Microbial Ecology

Bacterial Community of the Digestive Tract of the European Medicinal Leech ( Hirudo verbana ) from the Danube River
The original published version of this article had mistakes in figure legends. Correct figure legends are presented below.
“ Candidatus Hafkinia simulans” gen. nov., sp. nov., a Novel Holospora -Like Bacterium from the Macronucleus of the Rare Brackish Water Ciliate Frontonia salmastra (Oligohymenophorea, Ciliophora): Multidisciplinary Characterization of the New Endosymbiont and Its Host

Abstract

We characterized a novel Holospora-like bacterium (HLB) (Alphaproteobacteria, Holosporales) living in the macronucleus of the brackish water ciliate Frontonia salmastra. This bacterium was morphologically and ultrastructurally investigated, and its life cycle and infection capabilities were described. We also obtained its 16S rRNA gene sequence and performed in situ hybridization experiments with a specifically-designed probe. A new taxon, “Candidatus Hafkinia simulans”, was established for this HLB. The phylogeny of the family Holosporaceae based on 16S rRNA gene sequences was inferred, adding to the already available data both the sequence of the novel bacterium and those of other Holospora and HLB species recently characterized. Our phylogenetic analysis provided molecular support for the monophyly of HLBs and placed the new endosymbiont as the sister genus of Holospora. Additionally, the host ciliate F. salmastra, recorded in Europe for the first time, was concurrently described through a multidisciplinary study. Frontonia salmastra’s phylogenetic position in the subclass Peniculia and the genus Frontonia was assessed according to 18S rRNA gene sequencing. Comments on the biodiversity of this genus were added according to past and recent literature.

Soil Properties and Multi-Pollution Affect Taxonomic and Functional Bacterial Diversity in a Range of French Soils Displaying an Anthropisation Gradient

Abstract

The intensive industrial activities of the twentieth century have left behind highly contaminated wasteland soils. It is well known that soil parameters and the presence of pollutants shape microbial communities. But in such industrial waste sites, the soil multi-contamination with organic (polycyclic aromatic hydrocarbons, PAH) and metallic (Zn, Pb, Cd) pollutants and long-term exposure may induce a selection pressure on microbial communities that may modify soil functioning. The aim of our study was to evaluate the impact of long-term multi-contamination and soil characteristics on bacterial taxonomic and functional diversity as related to the carbon cycle. We worked on 10 soils from northeast of France distributed into three groups (low anthropised controls, slag heaps, and settling ponds) based on their physico-chemical properties (texture, C, N) and pollution level. We assessed bacterial taxonomic diversity by 16S rDNA Illumina sequencing, and functional diversity using Biolog® and MicroResp™ microtiter plate tools. Although taxonomic diversity at the phylum level was not different among the soil groups, many operational taxonomic units were influenced by metal or PAH pollution, and by soil texture and total nitrogen content. Functional diversity was not influenced by PAH contamination while metal pollution selected microbial communities with reduced metabolic functional diversity but more tolerant to zinc. Limited microbial utilisation of carbon substrates in metal-polluted soils was mainly due to the nitrogen content. Based on these two observations, we hypothesised that reduced microbial activity and lower carbon cycle–related functional diversity may have contributed to the accumulation of organic matter in the soils that exhibited the highest levels of metal pollution.

Bacterial Community of the Digestive Tract of the European Medicinal Leech ( Hirudo verbana ) from the Danube River

Abstract

The digestive tract of medicinal leeches from commercial suppliers has been investigated previously and comprises of a relatively simple bacterial community. However, the microbiome of medicinal leeches collected directly from the natural habitat has not been examined. In this study, we characterized the bacterial community in the digestive tract (anterior crop, posterior crop, and intestine) of the European medicinal leech, Hirudo verbana, collected from the Danube river using culture-independent and culture-dependent approaches. Culture-independent approach confirmed that the digestive tract of H. verbana carries a relatively simple bacterial community with species richness in the individual samples ranging from 43 to164. The dominant bacterial taxon was Mucinivorans sp. (49.7% of total reads), followed by Aeromonas sp. (18.7% of total reads). Several low abundance taxa, new for H. verbana, such as Phreatobacter, Taibaiella, Fluviicola, Aquabacterium, Burkholderia, Hydrogenophaga, Wolinella, and unidentified Chitinophagia, were also detected. The aerobic culturing approach showed Aeromonas veronii (Proteobacteria), the known leech symbiont, as the most dominant taxon followed by several Pseudomonas and Acidovorax spp. No significant differences in the bacterial community composition were detected among different parts of the digestive tract of individual leeches. However, the overall composition of the bacterial community among individual specimen varied significantly and this is possibly due to differences in leech age, feeding status, and blood source. Our results showed that the core bacterial community of H. verbana collected from the natural habitat is similar to that reported from the digestive tract of commercially supplied leeches maintained in the laboratory.

Experimental Testing of Dispersal Limitation in Soil Bacterial Communities with a Propagule Addition Approach

Abstract

The role of dispersal in the assembly of microbial communities remains contentious. This study tested the importance of dispersal limitation for the structuring of local soil bacterial communities using an experimental approach of propagule addition. Microbes extracted from soil pooled from samples collected at 20 localities across ~ 400 km in a temperate steppe were added to microcosms of local soils at three sites; the microcosms were then incubated in situ for 3 months. We then assessed the composition and diversity of bacterial taxa in the soils using 16S rRNA gene amplicon sequencing. The addition of the regional microbial pool did not cause significant changes in the overall composition or diversity of the total bacterial community, although a very small number of individual taxa may have been affected by the addition treatment. Our results suggest a negligible role of dispersal limitation in structuring soil bacterial communities in our study area.

Early Changes in Nutritional Conditions Affect Formation of Synthetic Mutualism Between Chlorella sorokiniana and the Bacterium Azospirillum brasilense

Abstract

The effect of three different nutritional conditions during the initial 12 h of interaction between the microalgae Chlorella sorokiniana UTEX 2714 and the plant growth–promoting bacterium Azospirillum brasilense Cd on formation of synthetic mutualism was assessed by changes in population growth, production of signal molecules tryptophan and indole-3-acetic acid, starch accumulation, and patterns of cell aggregation. When the interaction was supported by a nutrient-rich medium, production of both signal molecules was detected, but not when this interaction began with nitrogen-free (N-free) or carbon-free (C-free) media. Overall, populations of bacteria and microalgae were larger when co-immobilized. However, the highest starch production was measured in C. sorokiniana immobilized alone and growing continuously in a C-free mineral medium. In this interaction, the initial nutritional condition influenced the time at which the highest accumulation of starch occurred in Chlorella, where the N-free medium induced faster starch production and the richer medium delayed its accumulation. Formation of aggregates made of microalgae and bacteria occurred in all nutritional conditions, with maximum at 83 h in mineral medium, and coincided with declining starch content. This study demonstrates that synthetic mutualism between C. sorokiniana and A. brasilense can be modulated by the initial nutritional condition, mainly by the presence or absence of nitrogen and carbon in the medium in which they are interacting.

Genetic and Genome Analyses Reveal Genetically Distinct Populations of the Bee Pathogen Nosema ceranae from Thailand

Abstract

The recent global decline in Western honeybee (Apis mellifera) populations is of great concern for pollination and honey production worldwide. Declining honeybee populations are frequently infected by the microsporidian pathogen Nosema ceranae. This species was originally described in the Asiatic honeybee (Apis cerana), and its identification in global A. mellifera hives could result from a recent host transfer. Recent genome studies have found that global populations of this parasite are polyploid and that humans may have fueled their global expansion. To better understand N. ceranae biology, we investigated its genetic diversity within part of their native range (Thailand) and among different hosts (A. mellifera, A. cerana) using both PCR and genome-based methods. We find that Thai N. ceranae populations share many SNPs with other global populations and appear to be clonal. However, in stark contrast with previous studies, we found that these populations also carry many SNPs not found elsewhere, indicating that these populations have evolved in their current geographic location for some time. Our genome analyses also indicate the potential presence of diploidy within Thai populations of N. ceranae.

Population and Culture Age Influence the Microbiome Profiles of House Dust Mites

Abstract

Interactions with microorganisms might enable house dust mites (HDMs) to derive nutrients from difficult-to-digest structural proteins and to flourish in human houses. We tested this hypothesis by investigating the effects of changes in the mite culture growth and population of two HDM species on HDM microbiome composition and fitness. Growing cultures of laboratory and industrial allergen-producing populations of Dermatophagoides farinae (DFL and DFT, respectively) and Dermatophagoides pteronyssinus (DPL and DPT, respectively) were sampled at four time points. The symbiotic microorganisms of the mites were characterized by DNA barcode sequencing and quantified by qPCR using universal/specific primers. The population growth of mites and nutrient contents of mite bodies were measured and correlated with the changes in bacteria in the HDM microbiome. The results showed that both the population and culture age significantly influenced the microbiome profiles. Cardinium formed 93% and 32% of the total sequences of the DFL and DFT bacterial microbiomes, respectively, but this bacterial species was less abundant in the DPL and DPT microbiomes. Staphylococcus abundance was positively correlated with increased glycogen contents in the bodies of mites, and increased abundances of Aspergillus, Candida, and Kocuria were correlated with increased lipid contents in the bodies of mites. The xerophilic fungus Wallemia accounted for 39% of the fungal sequences in the DPL microbiome, but its abundance was low in the DPT, DFL, and DFT microbiomes. With respect to the mite culture age, we made three important observations: the mite population growth from young cultures was 5–8-fold higher than that from old cultures; specimens from old cultures had greater abundances of fungi and bacteria in their bodies; and yeasts predominated in the gut contents of specimens from young cultures, whereas filamentous mycelium prevailed in specimens from old cultures. Our results are consistent with the hypothesis that mites derive nutrients through associations with microorganisms.

Geographic and Temporal Variation of Distinct Intracellular Endosymbiont Strains of Wolbachia sp. in the Grasshopper Chorthippus parallelus : a Frequency-Dependent Mechanism?

Abstract

Wolbachia is an intracellular endosymbiont that can produce a range of effects on host fitness, but the temporal dynamics of Wolbachia strains have rarely been experimentally evaluated. We compare interannual strain frequencies along a geographical region for understanding the forces that shape Wolbachia strain frequency in natural populations of its host, Chorthippus parallelus (Orthoptera, Acrididae). General linear models show that strain frequency changes significantly across geographical and temporal scales. Computer simulation allows to reject the compatibility of the observed patterns with either genetic drift or sampling errors. We use consecutive years to estimate total Wolbachia strain fitness. Our estimation of Wolbachia fitness is significant in most cases, within locality and between consecutive years, following a negatively frequency-dependent trend. Wolbachia spp. B and F strains show a temporal pattern of variation that is compatible with a negative frequency-dependent natural selection mechanism. Our results suggest that such a mechanism should be at least considered in future experimental and theoretical research strategies that attempt to understand Wolbachia biodiversity.

Titanium Ions Inhibit the Bacteria in Vase Solutions of Freshly Cut Gerbera jamesonii and Extend the Flower Longevity

Abstract

Titanium ions significantly promote plant growth, but the mechanism is still unclear. Cut flowers are ideal materials for the study of plant growth and senescence. In this study, freshly cut Gerbera jamesonii were used to study the effects of titanium ions (8 mg/L) on the flower longevity. Flowering observation showed that the gerbera vase life was significantly prolonged in the presence of titanium ions. Plate colony counts showed that the amounts of bacteria in the vase solution of the control group were approximately 1700 times more than that of titanium ion treatment group. High-throughput sequencing was used to determine the sequences of 16S rRNA gene V3-V4 variable regions of the vase solutions to analyze bacterial species, their average proportions, and absolute abundance. The results showed that the titanium ions reduced the entire bacterial counts as well as altered the absolute abundance of different bacterial species in the vase solution. The most prevalent bacteria were mainly Enterobacteriaceae, Pseudomonas veronii, Pseudomonas sp., Delftia sp., Agrobacterium sp., Sphingobacterium multivorum, Acinetobacter johnsonii, and Clostridiaceae. In combination with plate colony counts, we demonstrated that all the bacterial growths were significantly inhibited by titanium ions, regardless of their average proportions increased or decreased. These results showed that titanium ions could extend effectively the longevity of gerberas and possess the broad-spectrum antibacterial properties. This study provides a basis for further mechanism exploration of titanium ions action and its applications in cut flower preservation and agricultural production.

Microbial Ecology

Cellular and Molecular

From neural crest cells to melanocytes: cellular plasticity during development and beyond

Abstract

Here, we review melanocyte development and how the embryonic melanoblast, although specified to become a melanocyte, is prone to cellular plasticity and is not fully committed to the melanocyte lineage. Even fully differentiated and pigment-producing melanocytes do not always have a stable phenotype. The gradual lineage restriction of neural crest cells toward the melanocyte lineage is determined by both cell-intrinsic and extracellular signals in which differentiation and pathfinding ability reciprocally influence each other. These signals are leveraged by subtle differences in timing and axial positioning. The most extensively studied migration route is the dorsolateral path between the dermomyotome and the prospective epidermis, restricted to melanoblasts. In addition, the embryonic origin of the skin dermis through which neural crest derivatives migrate may also affect the segregation between melanogenic and neurogenic cells in embryos. It is widely accepted that, irrespective of the model organism studied, the immediate precursor of both melanoblast and neurogenic populations is a glial-melanogenic bipotent progenitor. Upon exposure to different conditions, melanoblasts may differentiate into other neural crest-derived lineages such as neuronal cells and vice versa. Key factors that regulate melanoblast migration and patterning will regulate melanocyte homeostasis during different stages of hair cycling in postnatal hair follicles.

N -acetylglucosaminyltransferases and nucleotide sugar transporters form multi-enzyme–multi-transporter assemblies in golgi membranes in vivo

Abstract

Branching and processing of N-glycans in the medial-Golgi rely both on the transport of the donor UDP-N-acetylglucosamine (UDP-GlcNAc) to the Golgi lumen by the SLC35A3 nucleotide sugar transporter (NST) as well as on the addition of the GlcNAc residue to terminal mannoses in nascent N-glycans by several linkage-specific N-acetyl-glucosaminyltransferases (MGAT1-MGAT5). Previous data indicate that the MGATs and NSTs both form higher order assemblies in the Golgi membranes. Here, we investigate their specific and mutual interactions using high-throughput FRET- and BiFC-based interaction screens. We show that MGAT1, MGAT2, MGAT3, MGAT4B (but not MGAT5) and Golgi alpha-mannosidase IIX (MAN2A2) form several distinct molecular assemblies with each other and that the MAN2A2 acts as a central hub for the interactions. Similar assemblies were also detected between the NSTs SLC35A2, SLC35A3, and SLC35A4. Using in vivo BiFC-based FRET interaction screens, we also identified novel ternary complexes between the MGATs themselves or between the MGATs and the NSTs. These findings suggest that the MGATs and the NSTs self-assemble into multi-enzyme/multi-transporter complexes in the Golgi membranes in vivo to facilitate efficient synthesis of complex N-glycans.

Dgcr8 knockout approaches to understand microRNA functions in vitro and in vivo

Abstract

Biologic function of the majority of microRNAs (miRNAs) is still unknown. Uncovering the function of miRNAs is hurdled by redundancy among different miRNAs. The deletion of Dgcr8 leads to the deficiency in producing all canonical miRNAs, therefore, overcoming the redundancy issue. Dgcr8 knockout strategy has been instrumental in understanding the function of miRNAs in a variety of cells in vitro and in vivo. In this review, we will first give a brief introduction about miRNAs, miRNA biogenesis pathway and the role of Dgcr8 in miRNA biogenesis. We will then summarize studies performed with Dgcr8 knockout cell models with a focus on embryonic stem cells. After that, we will summarize results from various in vivo Dgcr8 knockout models. Given significant phenotypic differences in various tissues between Dgcr8 and Dicer knockout, we will also briefly review current progresses on understanding miRNA-independent functions of miRNA biogenesis factors. Finally, we will discuss the potential use of a new strategy to stably express miRNAs in Dgcr8 knockout cells. In future, Dgcr8knockout approaches coupled with innovations in miRNA rescue strategy may provide further insights into miRNA functions in vitro and in vivo.

Establishment and depletion of the ovarian reserve: physiology and impact of environmental chemicals

Abstract

The reproductive life span in women starts at puberty and ends at menopause, following the exhaustion of the follicle stockpile termed the ovarian reserve. Increasing data from experimental animal models and epidemiological studies indicate that exposure to a number of ubiquitously distributed reproductively toxic environmental chemicals (RTECs) can contribute to earlier menopause and even premature ovarian failure. However, the causative relationship between environmental chemical exposure and earlier menopause in women remains poorly understood. The present work, is an attempt to review the current evidence regarding the effects of RTECs on the main ovarian activities in mammals, focusing on how such compounds can affect the ovarian reserve at any stages of ovarian development. We found that in rodents, strong evidence exists that in utero, neonatal, prepubescent and even adult exposure to RTECs leads to impaired functioning of the ovary and a shortening of the reproductive lifespan. Regarding human, data from cross-sectional surveys suggest that human exposure to certain environmental chemicals can compromise a woman’s reproductive health and in some cases, correlate with earlier menopause. In conclusion, evidences exist that exposure to RTECs can compromise a woman’s reproductive health. However, human exposures may date back to the developmental stage, while the adverse effects are usually diagnosed decades later, thus making it difficult to determine the association between RTECs exposure and human reproductive health. Therefore, epidemiological surveys and more experimental investigation on humans, or alternatively primates, are needed to determine the direct and indirect effects caused by RTECs exposure on the ovary function, and to characterize their action mechanisms.

Exosomes: from carcinogenesis and metastasis to diagnosis and treatment of gastric cancer

Abstract

Exosomes represent an important group of extracellular vesicles with a defined size between 40 and 150 nm and cup-shaped construction which have a pivotal role in elimination of intracellular debris and intercellular signaling networks. A line of evidence revealed the impact of different types of exosomes in initiation, progression, and metastasis of gastric cancer (GC). These bioactive vesicles mediate tumor and stromal communication network through modulation of cell signaling for carcinogenesis and pre-metastatic niche formation in distant organs. Exosomes contain various cargos including DNAs (mitochondrial and genomic), proteins, transposable elements, and RNAs (coding and noncoding) with different compositions related to functional status of origin cells. In this review, we summarize the main roles of key exosomal cargos in induction of exosome-mediated signaling in cancer cells. Body fluids are employed frequently as the source of exosomes released by tumor cells with a potential role in early diagnosis of GC and chemoresistance. These vesicles as non-toxic and non-immunogenic carriers are also found to be applied for novel drug delivery systems.

Non-obesogenic doses of fatty acids modulate the functionality of the circadian clock in the liver

Abstract

Saturated fatty acids, such as palmitate, lead to circadian disruption in cell culture. Moreover, information regarding the effects of unsaturated fatty acids on circadian parameters is scarce. We aimed at studying the effects of low doses of saturated as well as unsaturated fatty acids on circadian metabolism in vivo and at deciphering the mechanism by which fatty acids convey their effect. Mice were fed non-obesogenic doses of palm or olive oil and hepatocytes were treated with palmitate and oleate. Mice fed non-obesogenic doses of palm oil showed increased signaling towards fatty acid synthesis, while olive oil increased signaling towards fatty acid oxidation. Low doses of palmitate and oleate were sufficient to alter circadian rhythms, due to changes in the expression and/or activity of key metabolic proteins. Palmitate, but not oleate, counteracted the reduction in lipid accumulation and BMAL1-induced expression of mitochondrial genes involved in fatty acid oxidation. Palmitate was also found to interfere with the transcriptional activity of CLOCK:BMAL1 by preventing BMAL1 deacetylation and activation. In addition, palmitate, but not oleate, reduced PER2-mediated transcriptional activation and increased REV-ERBα-mediated transcriptional inhibition of Bmal1. The inhibition of PER2-mediated transcriptional activation by palmitate was achieved by interfering with PER2 nuclear translocation. Indeed, PER2 reduced fat accumulation in hepatocytes and this reduction was prevented by palmitate. Herein, we show that the detrimental metabolic alteration seen with high doses of palmitate manifests itself early on even with non-obesogenic levels. This is achieved by modulating BMAL1 at several levels abrogating its activity and expression.

Establishment and maintenance of blood–lymph separation

Abstract

Hippocratic Corpus, a collection of Greek medical literature, described the functional anatomy of the lymphatic system in the fifth century B.C. Subsequent studies in cadavers and surgical patients firmly established that lymphatic vessels drain extravasated interstitial fluid, also known as lymph, into the venous system at the bilateral lymphovenous junctions. Recent advances revealed that lymphovenous valves and platelet-mediated hemostasis at the lymphovenous junctions maintain life-long separation of the blood and lymphatic vascular systems. Here, we review murine models that exhibit failure of blood–lymph separation to highlight the novel mechanisms and molecular targets for the modulation of lymphatic disorders. Specifically, we focus on the transcription factors, cofactors, and signaling pathways that regulate lymphovenous valve development and platelet-mediated lymphovenous hemostasis, which cooperate to maintain blood–lymph separation.

Therapeutic potential of menstrual blood-derived endometrial stem cells in cardiac diseases

Abstract

Despite significant developments in medical and surgical strategies, cardiac diseases remain the leading causes of morbidity and mortality worldwide. Numerous studies involving preclinical and clinical trials have confirmed that stem cell transplantation can help improve cardiac function and regenerate damaged cardiac tissue, and stem cells isolated from bone marrow, heart tissue, adipose tissue and umbilical cord are the primary candidates for transplantation. During the past decade, menstrual blood-derived endometrial stem cells (MenSCs) have gradually become a promising alternative for stem cell-based therapy due to their comprehensive advantages, which include their ability to be periodically and non-invasively collected, their abundant source material, their ability to be regularly donated, their superior proliferative capacity and their ability to be used for autologous transplantation. MenSCs have shown positive therapeutic potential for the treatment of various diseases. Therefore, aside from a brief introduction of the biological characteristics of MenSCs, this review focuses on the progress being made in evaluating the functional improvement of damaged cardiac tissue after MenSC transplantation through preclinical and clinical studies. Based on published reports, we conclude that the paracrine effect, transdifferentiation and immunomodulation by MenSC promote both regeneration of damaged myocardium and improvement of cardiac function.

Presynaptic NMDA receptors control nociceptive transmission at the spinal cord level in neuropathic pain

Abstract

Chronic neuropathic pain is a debilitating condition that remains challenging to treat. Glutamate N-methyl-d-aspartate receptor (NMDAR) antagonists have been used to treat neuropathic pain, but the exact sites of their actions have been unclear until recently. Although conventionally postsynaptic, NMDARs are also expressed presynaptically, particularly at the central terminals of primary sensory neurons, in the spinal dorsal horn. However, presynaptic NMDARs in the spinal cord are normally quiescent and are not actively involved in physiological nociceptive transmission. In this review, we describe the emerging role of presynaptic NMDARs at the spinal cord level in chronic neuropathic pain and the implications of molecular mechanisms for more effective treatment. Recent studies indicate that presynaptic NMDAR activity at the spinal cord level is increased in several neuropathic pain conditions but not in chronic inflammatory pain. Increased presynaptic NMDAR activity can potentiate glutamate release from primary afferent terminals to spinal dorsal horn neurons, which is crucial for the synaptic plasticity associated with neuropathic pain caused by traumatic nerve injury and chemotherapy-induced peripheral neuropathy. Furthermore, α2δ-1, previously considered a calcium channel subunit, can directly interact with NMDARs through its C-terminus to increase presynaptic NMDAR activity by facilitating synaptic trafficking of α2δ-1–NMDAR complexes in neuropathic pain caused by chemotherapeutic agents and peripheral nerve injury. Targeting α2δ-1–bound NMDARs with gabapentinoids or α2δ-1 C-terminus peptides can attenuate nociceptive drive form primary sensory nerves to dorsal horn neurons in neuropathic pain.

Tight junction proteins at the blood–brain barrier: far more than claudin-5

Abstract

At the blood–brain barrier (BBB), claudin (Cldn)-5 is thought to be the dominant tight junction (TJ) protein, with minor contributions from Cldn3 and -12, and occludin. However, the BBB appears ultrastructurally normal in Cldn5 knock-out mice, suggesting that further Cldns and/or TJ-associated marvel proteins (TAMPs) are involved. Microdissected human and murine brain capillaries, quickly frozen to recapitulate the in vivo situation, showed high transcript expression of Cldn5, -11, -12, and -25, and occludin, but also abundant levels of Cldn1 and -27 in man. Protein levels were quantified by a novel epitope dilution assay and confirmed the respective mRNA data. In contrast to the in vivo situation, Cldn5 dominates BBB expression in vitro, since all other TJ proteins are at comparably low levels or are not expressed. Cldn11 was highly abundant in vivo and contributed to paracellular tightness by homophilic oligomerization, but almost disappeared in vitro. Cldn25, also found at high levels, neither tightened the paracellular barrier nor interconnected opposing cells, but contributed to proper TJ strand morphology. Pathological conditions (in vivo ischemia and in vitro hypoxia) down-regulated Cldn1, -3, and -12, and occludin in cerebral capillaries, which was paralleled by up-regulation of Cldn5 after middle cerebral artery occlusion in rats. Cldn1 expression increased after Cldn5 knock-down. In conclusion, this complete Cldn/TAMP profile demonstrates the presence of up to a dozen TJ proteins in brain capillaries. Mouse and human share a similar and complex TJ profile in vivo, but this complexity is widely lost under in vitro conditions.

Cellular and Molecular

Parasitology

Apparent kleptoparasitism in fish—parasitic gnathiid isopods” 10.1007/s00436-018-6152-8
Correction to: Epidemiology of Sulcascaris sulcata (Nematoda: Anisakidae) ulcerous gastritis in the Mediterranean loggerhead sea turtle ( Caretta caretta )
The original version of this article contained a mistake in the value of Bar in figure 3 caption. It should be Bar = 200 μm instead of Bar = 500 μm.
Correction to: In vitro schistosomicidal activity of tamoxifen and its effectiveness in a murine model of schistosomiasis at a single dose
The original published version of this article contains error in Tables 1 and 2. Correct tables are presented here.
First report of sparganosis manifested by pleuritis and decreased peripheral blood eosinophils in Jiangsu province, China

Abstract

Sparganosis is a parasitic infection caused by the metacestode stage of Spirometra mansoni and some other related diphyllobothriidean cestodes. Although various internal organs were involved in sparganum infection, pulmonary and pleural involvement is rarely reported. We herein report an uncommon form of sparganosis manifested by pleuritis and decreased peripheral blood eosinophils. Sparganum worms were found in the pleural effusion accidentally and confirmed by pathological diagnosis. After being treated with praziquantel for 10 days, the patient’s symptoms, laboratory examinations, and imaging findings were improved gradually.

Distribution of Cryptosporidium parvum gp60 subtypes in calf herds of Saxony, Germany

Abstract

Cryptosporidiosis is a common protozoan parasitic infection that causes diarrhoea in neonatal calves. The high shedding of environmentally resistant oocysts facilitates outbreaks of cryptosporidiosis in humans. In total, 58 farms (512 calves) in Germany (Saxony and Brandenburg) were visited three times each. Faecal samples of pre-weaned calves were microscopically examined for oocysts of Cryptosporidium spp. using Heine staining and were scored with regard to their consistency. Overall, 88.9% of calves tested microscopically positive for Cryptosporidium spp. in at least one sample, and the excretion of oocysts was significantly (P < 0.01) associated with a higher faecal score (diarrhoea). After DNA extraction from pooled farm isolates, 47 samples were successfully subtyped by sequence analysis of the 60 kDa glycoprotein gene (gp60). All isolates belonged to subtype family IIa. IIaA15G2R1 was the most common subtype (present on 66% of the farms), followed by IIaA16G3R1 (13%). Subtypes IIaA14G1R1, IIaA14G2R1, IIaA1612R1, IIaA16G2R1, IIaA17G1R1, IIaA17G2R1, IIaA17G4R1 and IIaA19G2R1 were found sporadically. This is the first description of gp60 subtype IIaA17G4R1 in cattle in Germany.

Combined morphology and DNA-barcoding to identify Plagiorhynchus cylindraceus cystacanths in Atelerix algirus

Abstract

The acanthocephalan parasite Plagiorhynchus cylindraceus has a global distribution and utilizes isopods and birds as intermediate and definitive hosts, respectively. Occasionally, mammals of various orders can act as paratenic hosts. In hedgehogs, severe cases have been reported in juvenile specimens due to secondary infections, as a consequence of complete penetrations of the intestinal wall by cystacanths. In a 66-month study period, we found seven larvae of this parasite encysted in both, the peritoneal cavity and intestine of the Algerian hedgehog, Atelerix algirus in Majorca. Morphology alone was insufficient to identify the species, due to the lack of previous reports and taxonomy-informative characters. In the present report, we combined the use of morphology and the DNA-barcoding approach to confirm to identify cystacanths as P. cylindraceus. This is the first report of this parasite in this hedgehog species. The epidemiological implications will be discussed, including the risk of zoonosis and the importance of using modern approaches to identify immature acanthocephalan larvae in wildlife hosts.

Intestinal injury caused by Eimeria spp. impairs the phosphotransfer network and gain weight in experimentally infected chicken chicks

Abstract

Parasitic infections caused by protozoan belonging to genus Eimeria are considered important for the poultry industry, due to their severe intestinal lesions and high mortality rates, causing significant economic losses. Although several mechanisms of coccidiosis pathogenesis are known, the effects of this infection on intestinal enzymes linked to adenosine triphosphate (ATP) metabolism, as creatine kinase (CK), adenylate kinase (AK), and pyruvate kinase (PK), remain unknown. Thus, the aim of this study was to evaluate whether coccidiosis impairs enzymes linked ATP metabolism in the intestine of chicken chicks. For this, 42 animals that were 2 days old were divided into two groups: uninfected (the negative control group) and experimentally infected on second day of life (the positive control group). On days 5, 10, and 15 post-infection (PI), fecal samples were collected for oocyst counts; intestinal tissue was collected in order to evaluate CK, AK, and PK activities, as well as parameters of the oxidative stress and histopathology. On days 10 and 15 PI, infected animals showed high counts of oocysts in fecal samples and intestinal lesions compared to the control group. Cytosolic CK activity was higher in infected animals on days 10 and 15 PI compared to the control group, while mitochondrial CK activity was lower on days 5, 10, and 15 PI. Also, AK activity was lower in infected animals on days 10 and 15 PI compared to control group, while no differences were observed between groups regarding PK activity. In relation to parameters of oxidative stress, intestinal lipid peroxidation and reactive oxygen species levels were higher in infected animals on days 10 and 15 PI compared to the control group, while non-protein thiol levels were lower on day 10 PI. On the 15th day, infected animals had lower body weight (P < 0.05). Based on this evidence, inhibition of mitochondrial CK activity causes an impairment of intestinal energetic homeostasis possibly through depletion on ATP levels, although the cytosolic CK activity acted as an attempt to restore the mitochondrial ATP levels through a feedback mechanism. Moreover, the impairment on energy metabolism appears to be mediated by excessive production of intestinal ROS, as well as oxidation of lipids and thiol groups.

Oral infection of mice and host cell invasion by Trypanosoma cruzi strains from Mexico

Abstract

Oral infection by Trypanosoma cruzi has been responsible for frequent outbreaks of acute Chagas disease in the north of South America and in the Amazon region, where T. cruzi genetic group TcI predominates. TcI strains from different geographical regions have been used in oral infection in mice, but there is no information about strains from Mexico where TcI is prevalent. Here, we analyzed four Mexican strains as concerns the course of oral infection, the ability to invade host cells in vitro, and the profile of metacyclic trypomastigote surface molecules gp82 and gp90 that are implicated in parasite internalization. Oral infection of mice with metacyclic forms of all strains resulted in reduced blood and tissue parasitism, and mild to moderate inflammatory process in the heart/skeletal muscle. They expressed pepsin-resistant gp82 and gp90 molecules at high levels and invaded host cells poorly in full nutrient medium and efficiently under nutrient-deprived condition. The properties exhibited by Mexican strains were similar to those displayed by TcI strains from other geographical regions, reinforcing the notion that these features are common to the genetic group TcI as a whole.

Antarctophthirus microchir infestation in synanthropic South American sea lion ( Otaria flavescens ) males diagnosed by a novel non-invasive method

Abstract

Antarctophthirus microchir is a sucking louse species belonging to the family Echinophthiriidae and has been reported to parasitize all species of the subfamily Otariinae, the sea lions. Former studies on this ectoparasite mainly required fixation, immobilization, or death of host species and especially examinations of adult male sea lions are still very rare. Between March and May 2018, adult individuals of a unique “urban” bachelor group of South American sea lions (Otaria flavescens) living directly in the city of Valdivia, Chile, were studied regarding their ectoparasite infestation status. For first time, a non-invasive method in the form of a lice comb screwed on a telescopic rod and grounded with adhesive tape was used for sample taking process. Overall, during combing different stages of A. microchir were detected in 4/5 O. flavescens individuals, especially at the junction between the back and hind flippers. Our findings represent the first report of A. microchir infesting individuals of this synanthropic colony and fulfilling complete life cycle in a sea lion group despite inhabiting freshwater and in absence of females/pups. Our “telescopic lice comb apparatus” offers a new strategy to collect different stages of ectoparasites and a range of epidermal material, such as fur coat hair and superficial skin tissue for a broad spectrum of research fields in wildlife sciences in an unmolested and stress reduced manner.

On the occurrence and molecular identification of Contracaecum larvae (Nematoda: Anisakidae) in Mugil cephalus from Turkish waters

Abstract

Anisakis and Contracaecum species are fish borne zoonotic nematodes. In our previous studies, other larval anisakid and raphidascarid nematodes, Anisakis and Hysterothylacium species, were genetically identified in marine fish from Turkish waters. However, there is no information on molecular identification of larval Contracaecum species in marine fish from Turkey. Therefore, the aim of this study was only to investigate the presence and molecular identification of Contracaecum species in commonly commercialized marine fish from Turkish waters. A total of 475 marine fish, which belong to 21 different species, were sampled from the Aegean (FAO 37.3.1), Mediterranean (FAO 37.3.2), and Black Sea (FAO 37.4.2). The prevalence of Contracaecum L3 larvae in the Aegean Sea was identified as 10% in Mugil cephalus. All Contracaecum L3 larvae were molecularly characterized with RFLP targeting the ITS region and rrnS gene. Moreover, all larvae were analyzed by sequencing of ITS region, rrnS and cox2 gene. All Contracaecum larvae were identified as C. overstreeti based on the cox2 sequence analysis. This is the first report of C. overstreeti larvae in M. cephalus as paratenic and intermediate hosts. Furthermore, the analysis reveals novel information on ITS region. Additionally, the rrnS gene of C. overstreeti was also achieved and deposited in Genbank for the first time. The PCR-RFLP patterns of the ITS region and rrnS gene from C. overstreeti were presented in the present study. Consequently, the presence of C. overstreeti larvae in M. cephalus from the Aegean Sea may also potentially capable of inducing allergic sensitization in humans.

Parasitology

Voice

Vocal Health Education in Undergraduate Performing Arts Training Programs

Amanda Flynnlow asterisk,'Correspondence information about the author Amanda FlynnEmail the author Amanda Flynn
Pace University, New York, New York

Journal of Voice

Accepted: March 27, 2019; Published online: April 29, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.016
Publication stage: In Press Corrected Proof

SUMMARY

Purpose

Vocal health is taught in multiple formats and to varying degrees across undergraduate training programs. The aim of the study is to identify what methods of instruction lead to a better self-perception of vocal health in order to more adequately prepare graduates for the extreme demands of the performing arts industry.

Method

A survey investigating how vocal health and vocal injury are being taught was administered to students within 5 years of graduation. This study looked at what type of information is taught, when information is presented, how prepared students feel they are to handle their health and injuries upon graduation, the prevalence of injuries while in school and during the first 5 years of postgraduation, and students’ suggestions to improve their overall vocal health education. The survey compares vocal health education between the various disciplines of classical voice, musical theatre, and acting.

Results

Students learn more about general vocal hygiene than voice disorders in their undergraduate programs. Classical voice and musical theatre majors learn more about vocal health than acting majors, yet acting majors report a higher incidence of vocal injury within the first 5 years of graduation. Students also retain more vocal health knowledge when presented with information multiple times in their education.

Conclusions

Creating a more specific, consistent vocal health curriculum for all voice-related performing arts programs will improve self-efficacy regarding vocal health upon graduation.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
The Influence of Linguistic Demand on Symptom Expression in Adductor Spasmodic Dysphonia

Laura L.O. Froeschkelow asterisk,'Correspondence information about the author Laura L.O. FroeschkeEmail the author Laura L.O. Froeschke
Department of Communication Sciences and Disorders, Elmhurst College, Elmhurst, Illinois

Journal of Voice

Accepted: April 10, 2019; Published online: April 29, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.04.003
Publication stage: In Press Corrected Proof

Summary

Introduction

Adductor Spasmodic Dysphonia (ADSD), a form of focal dystonia, has been defined as a neurogenic, task-specific disorder characterized by abrupt spasms of intrinsic laryngeal muscles that result in phonatory breaks. Voice breaks are typically isolated to propositional speech, and reported to increase in severity as speaking demand or complexity increases. Research to date has focused on variations in phonologic contexts and their influence on voice breaks. The influences of variables at lexical and syntactic levels of analysis have been less well-researched and yet may provide insight into observed variability of symptom manifestation in this rare voice disorder.

Objectives

This study investigated frequency of voice breaks over 20 standard sentences in 38 individuals with ADSD according to linguistic complexity measures including lexical density and a four-level lexical frequency and type paradigm. Two research questions about linguistic influences and ADSD symptom manifestation were posed: (1) does the frequency of voice breaks vary according to the lexical density of a string? and (2) does the frequency of voice breaks vary according to a measure of lexical frequency/type?

Results

Results revealed a nonsignificant relationship between string length and voice break frequency, whereas a significant relationship was found between lexical density and voice break frequency (P = 0.029, r = 0.488). Lexical analysis results revealed a significant relationship between lexical frequency and voice breaks, with words within technical/academic classes relating to the highest rates of voice break across 38 subjects with ADSD.

Conclusions

Results from this secondary analysis provide support for the hypothesis that variation in linguistic demand may modulate symptom expression in SD. Specifically, lexical density and lexical frequency modulated the frequency of symptom expression in classic forms of SD in this purposive sample.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Effect of Dysphonia and Cognitive-Perceptual Listener Strategies on Speech Intelligibility

Connie K. Porcaro*,low asterisklow asterisk,'Correspondence information about the author Connie K. PorcaroEmail the author Connie K. Porcaro
, Paul M. Evitts†,‡
, Nicole King
, Cassandra Hood
, Erin Campbell
, Layla White
, Jacqueline Veraguas*

Journal of Voice

Accepted: March 21, 2019; Published online: April 25, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.013
Publication stage: In Press Corrected Proof

ABSTRACT

There is a high prevalence of dysphonia among professional voice users and the impact of the disordered voice on the speaker is well documented. However, there is minimal research on the impact of the disordered voice on the listener. Considering that professional voice users include teachers and air-traffic controllers, among others, it is imperative to determine the impact of a disordered voice on the listener. To address this, the objectives of the current study included: (1) determine whether there are differences in speech intelligibility between individuals with healthy voices and those with dysphonia; (2) understand whether cognitive-perceptual strategies increase speech intelligibility for dysphonic speakers; and (3) determine the relationship between subjective voice quality ratings and speech intelligibility. Sentence stimuli were recorded from 12 speakers with dysphonia and four age- and gender-matched typical, healthy speakers and presented to 129 healthy listeners divided into one of three strategy groups (ie, control, acknowledgement, and listener strategies). Four expert raters also completed a perceptual voice assessment using the Consensus Assessment Perceptual Evaluation of Voice for each speaker. Results indicated that dysphonic voices were significantly less intelligible than healthy voices (P ≤ 0.001) and the use of cognitive-perceptual strategies provided to the listener did not significantly improve speech intelligibility scores (P = 0.602). Using the subjective voice quality ratings, regression analysis found that breathiness was able to predict 41% of the variance associated with number of errors (P = 0.008). Overall results of the study suggest that speakers with dysphonia demonstrate reduced speech intelligibility and that providing the listener with specific strategies may not result in improved intelligibility.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Normative Value of SVHI-10. Systematic Review and Meta-Analysis

Maria Sobol, PhD *
, Ewelina M. Sielska-Badurek, Prof †,low asterisklow asterisk,'Correspondence information about the author Prof Ewelina M. Sielska-BadurekEmail the author Prof Ewelina M. Sielska-Badurek
, Anna Rzepakowska, Prof †
, Ewa Osuch-Wójcikiewicz, Prof †

Journal of Voice

Accepted: April 9, 2019; Published online: April 25, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.04.002
Publication stage: In Press Corrected Proof

Summary

Objective

The study aimed to determine the normative value of SVHI-10.

Study Design

The study is a systematic review and a meta-analysis.

Methods

A systematic literature search was performed using PubMed and ScienceDirect to access relevant databases and to locate outcome studies. Eligibility criteria included type of publication, participant characteristics, and report of outcomes. Data analysis was conducted using the meta-analysis method.

Results

Six articles were included for the final analysis. The normative values for the SVHI-10 for a group of 528 subjects were 8.38 with confidence levels of 7.43–9.34 (age range 16–83).

Conclusions

Based on the results of the meta-analysis the SVHI-10 can be used as a screening tool for a group of singers. In the future, it would be worthwhile to carry out a subordinate analysis to determine the SVHI-10 range for mild voice disorders or severe voice disorders in singing.

© 2019 Published by Elsevier Inc.

Article in Press
The Effectiveness of Vocal Hygiene Education for Decreasing At-Risk Vocal Behaviors in Vocal Performers

Fran Pomaville*,low asterisklow asterisk,'Correspondence information about the author Fran PomavilleEmail the author Fran Pomaville
, Kristi Tekerlek*
, Anthony Radford*

Journal of Voice

Accepted: March 12, 2019; Published online: April 25, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.004
Publication stage: In Press Corrected Proof

Summary

This study examined the knowledge gained and behavioral changes made by vocal performers after attending a vocal hygiene education program. A single-group, pretest-posttest research design was utilized to examine the improvement of voice care knowledge and decrease of phonotraumatic behaviors in vocal performers. Data analysis involved a comparison of pretest and posttest responses from an online questionnaire. A paired sample t test revealed a statistically significant improvement in the participants’ knowledge regarding the larynx, voice production, and vocal hygiene. In addition, a behavioral inventory revealed improved hydration habits, decreased caffeine and alcohol intake, and healthier responses to symptoms of throat irritation or vocal fatigue. The findings from this study will contribute to evidence regarding the benefits of having vocal performers attend a vocal hygiene education program in order to increase their knowledge about voice production and care, decrease at-risk vocal behaviors, and improve healthy vocal practices.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Morphology, Vibratory Function, and Vascular Pattern for Predicting Malignancy in Vocal Fold Leukoplakia

Anna Rzepakowska*,a,'Correspondence information about the author Anna RzepakowskaEmail the author Anna Rzepakowska
, Maria Sobol
, Ewelina Sielska-Badurek*
, Kazimierz Niemczyk*
, Ewa Osuch-Wójcikiewicz*

Journal of Voice

Accepted: April 2, 2019; Published online: April 19, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.04.001
Publication stage: In Press Corrected Proof

Summary

Background

The study evaluates clinical features of vocal fold (VF) leukoplakia in predicting its benign or malignant nature.

Materials

57 patients with 84 lesions were evaluated before undergoing laryngeal microsurgery. The texture, color, thickness and size of the leukoplakia, along with an assessment of the surrounding mucosa vascularization using narrow-band imaging (NBI), and VF vibratory function were analyzed. Receiver-operating characteristic curves were constructed to determine the predictive value of each feature and area under the curve (AUC) was calculated.

Results

Histopathological examination revealed high-grade dysplasia or invasive cancer in 13 of VF leukoplakia. Seventy-one lesions were nondysplastic or low-grade dysplasia. Nonhomogenous color, irregular texture, and prominent thickness predicted malignancy with statistical significance (P < 0.05). AUC was 0.793, 0.793, and 0.679, respectively. Absence of a mucosal wave on laryngovideostroboscopy was significant for the detection of malignancy (P < 0.001) with an AUC of 0.927. The NBI diagnosis of horizontal vessel loops was significant with the highest AUC of 0.993.

Conclusions

The comprehensive clinical evaluation of VF leukoplakia with laryngovideostroboscopy and NBI creates the opportunity to differentiate between low- and high-risk malignancy lesions. The perpendicular vascular pattern and the limited or absent mucosal wave appear to be the most powerful indicators of malignancy.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Acoustic Voice Quality Index and Acoustic Breathiness Index: Analysis With Different Speech Material in the Brazilian Portuguese

Marina Englert*,†,a,'Correspondence information about the author Marina EnglertEmail the author Marina Englert
, Livia Lima
, Mara Behlau*,†

Journal of Voice

Accepted: March 25, 2019; Published online: April 17, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.015
Publication stage: In Press Corrected Proof

Summary

Objective

To analyze the Acoustic Voice Quality Index (AVQI) and the Acoustic Breathiness Index (ABI) concurrent validity and diagnostic accuracy with different speech materials.

Methods

Voices of 53 subjects (40 dysphonic; 13 vocally health) were recorded: vowel /a/ + counting numbers 1–20 (42 syllables) + reading text (138 syllables). Numbers and text were edited in order to achieve 3 seconds of voiced segments, such as the vowel /a/ (average of 18.81 and 32.49 syllables; confidence interval of 1.87 and 2.30). The audio files were edited to have 17 syllables for numbers and 32 for text. Three voice specialists perceptually judge the overall voice quality (G) and the breathiness (B). AVQI’s and ABI’s precision and concurrent validity were assessed.

Results

The intra- and inter-rater reliability were high. Reading text presented higher concurrent validity (r) than automatic speech and excellent area under the receiver-operating characteristic curve for AVQI (0.963) and ABI (0.929). Counting numbers presented good area under the receiver-operating characteristic curve for AVQI (0.870) and excellent for ABI (0.924). Counting numbers produced higher sensitivity for ABI (95.2%) and reading text higher specificity for both indexes (AVQI = 100%; ABI = 90.90%). Reading text presented higher AVQI and ABI scores than numbers, therefore, reading seems to reveal more vocal deviations; however, perceptual judgment can be similar in both samples.

Conclusions

Different speech materials may impact acoustic outcomes and certain voice characteristics may not be evident. Reading text offers higher diagnostic accuracy. Clinician and/or researchers must select and standardize the speech sample according to their goals.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Efficacy of Videostroboscopy and High-Speed Videoendoscopy to Obtain Functional Outcomes From Perioperative Ratings in Patients With Vocal Fold Mass Lesions

Maria E. Powelllow asterisk,†,‡,a,'Correspondence information about the author Maria E. PowellEmail the author Maria E. Powell
, Dimitar D. Deliyski†,‡,§
, Steven M. Zeitels‖,¶
, James A. Burns‖,¶
, Robert E. Hillman‖,¶,#
, Terri Treman Gerlachlow asterisklow asterisk
, Daryush D. Mehta‖,¶,#

Journal of Voice

Accepted: March 21, 2019; Published online: April 17, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.012
Publication stage: In Press Corrected Proof

Summary

Objectives

A major limitation of comparing the efficacy of videostroboscopy (VS) and high-speed videoendoscopy (HSV) is the lack of an objective reference by which to compare the functional assessment ratings of the two techniques. For patients with vocal fold mass lesions, intraoperative measures of lesion size and depth may serve as this objective reference. This study compared the relationships between the pre- to postoperative change in VS and HSV visual-perceptual ratings to intraoperative measures of lesion size and depth.

Design

Prospective visual-perceptual study with intraoperative measures of lesion size and depth.

Methods

VS and HSV samples were obtained preoperatively and postoperatively from 28 patients with vocal fold lesions and from 17 vocally healthy controls. Two experienced clinicians rated amplitude, mucosal wave, vertical phase difference, left-right phase asymmetry, and vocal fold edge on a visual-analog scale using both imaging techniques. The change in perioperative ratings from VS and HSV was compared between groups and correlated to intraoperative measures of lesion size and depth.

Results

HSV was as reliable as VS for ratings of amplitude and edge, and substantially more reliable for ratings of mucosal wave and left-right phase asymmetry. Both VS and HSV had mild-moderate correlations between change in perioperative ratings and intraoperative measures of lesion area. Change in function could be obtained in more patients and for more parameters using HSV than VS. Group differences were noted for postoperative ratings of amplitude and edge; however, these differences were within one level of the visual-perceptual rating scale. The presence of asynchronicity in VS recordings renders vibratory features either uninterpretable or potentially distorted and thus should not be rated.

Conclusions

Amplitude and edge are robust vibratory measures for perioperative functional assessment, regardless of imaging modality. HSV is indicated for evaluation of subepithelial lesions or if asynchronicity is present in the VS image sequence.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
A Very Rare Complication of Hyaluronic Acid Injection for Medialization Laryngoplasty: A Case With Laryngeal Abscess

Necati Enverlow asterisk,'Correspondence information about the author Necati EnverEmail the author Necati Enver
, Orhan Asya
, Ghazi Abuzaid
, Ece Gürol
Department of Otolaryngology, Pendik Training and Research Hospital, Marmara University, Istanbul, Turkey

Journal of Voice

Accepted: March 21, 2019; Published online: April 17, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.011
Publication stage: In Press Corrected Proof

Summary

Hyaluronic acid injection for medialization laryngoplasty is a safe procedure performed on patients with glottic incompetence. Laryngeal abscess formation as a complication of injection laryngoplasty is a very rare complication, and, as we know from the literature, there has been only one case of laryngeal abscess after injection laryngoplasty in a patient with a type-I laryngeal cleft. We document for the first time a laryngeal abscess resulting from hyaluronic acid injection laryngoplasty for unilateral vocal fold paralysis. Prompt evaluation of the patient was necessary. Our patient was treated with antibiotics and corticosteroids without a need for intubation. One year after injection, the patient’s Voice Handicap Index-10 score was still good and within the range of normal values.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Association Between Subglottic Pressure and Pulmonary Function in Individuals With Parkinson’s Disease

Adrián Castillo*
, Javiera Castillo
, Alvaro Reyes‡,a,'Correspondence information about the author Alvaro ReyesEmail the author Alvaro Reyes

Journal of Voice

Accepted: March 5, 2019; Published online: April 15, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.001
Publication stage: In Press Corrected Proof

Summary

Introduction

In individuals with Parkinson’s disease (PD), pulmonary complication such as weakness and rigidity of respiratory muscles and reduced cough airflow may be associated with reduced voice production due to limited pulmonary capacity and reduced airflow needed to vibrate the vocal folds. It is not clear, however, which pulmonary function parameter is determinant in the association with peak subglottic pressure (SGP). Therefore, the purpose of this study was to determine the association between peak SGP and pulmonary function parameters in individuals with PD.

Methods

Forty-two individuals with diagnosis of idiopathic PD of both genders were recruited in the study. Mean and peak SGP, spirometric indices, maximum inspiratory pressure, maximum expiratory pressure (MEP), and peak cough flow (PCF) during reflex and voluntary cough were measured on all participants.

Results

The analysis revealed that peak SGP had a moderate but significant linear association with MEP (r = 0.38; P = 0.013), voluntary (r = 0.31; P= 0.051), and reflex PCF (r = 0.40; P = 0.012), but not with maximum inspiratory pressure (r = 0.23; P = 0.145). Higher values in peak SGP were associated with higher values in MEP, voluntary PCF, and reflex PCF. No linear association was detected between peak SGP and spirometric indices.

Conclusions

Peak SGP has a direct association with voluntary and reflex PCF, and expiratory muscle strength, but not with inspiratory muscle strength. The association with peak SGP is higher for reflex PCF than for voluntary PCF.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Longitudinal Case Study of Transgender Voice Changes Under Testosterone Hormone Therapy

Gabriel J. Cler*,†,low asterisklow asterisk,'Correspondence information about the author Gabriel J. ClerEmail the author Gabriel J. Cler
, Victoria S. McKenna
, Kimberly L. Dahl
, Cara E. Stepp*,†,‡,║

Journal of Voice

Accepted: March 14, 2019; Published online: April 13, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.006
Publication stage: In Press Corrected Proof

Abstract

The purpose of this study was to comprehensively evaluate voice and speech changes in one healthy 30-year-old transgender male undergoing testosterone therapy for transition. Testing occurred at three timepoints before cross-sex hormone therapy and every 2 weeks thereafter for 1 year. Data collected included measures of acoustics, aerodynamics, and laryngeal structure and function via flexible laryngoscopy. Analysis included acoustic correlates of pitch, loudness, voice quality, and vocal tract length, as well as perceptual measures of voice quality and gender. Speaking fundamental frequency (fo) lowered from 183 Hz to 134 Hz. Phonatory frequency range (ie, minimum and maximum singing range) shifted from a range of D#3–E6 to a range of A2–A5. Perceptual measures of voice quality indicated no negative changes. Naïve listeners reliably rated the participant’s speech samples as male after 37 weeks on testosterone. Few studies document in detail the variety of voice changes that occur during cross-sex hormone therapy, focusing instead on fo alone. This study adds to the literature a comprehensive case study of speech and voice changes experienced by one transmasculine participant undergoing testosterone therapy.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Air Injection and Transillumination in Phonosurgery: A Novel Technique

Ismail Kocak*
, Zeynep Alkan†,low asterisklow asterisk,'Correspondence information about the author Zeynep AlkanEmail the author Zeynep Alkan
, Okan Ovunc

Journal of Voice

Accepted: March 20, 2019; Published online: April 12, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.009
Publication stage: In Press Corrected Proof

Abstract

Objectives/Hypothesis

The goal of this study was to introduce two novel techniques in phonomicrosurgery, air injection (AIR), and transillumination (TI), to improve the diagnosis and surgical excision of pathological tissue in vocal folds during suspension laryngoscopy while preserving the healthy tissue as much as possible.

Study design

Prospective clinical case series.

Methods

Thirty-four patients with benign vocal cord lesions who underwent phonomicrosurgery between January 2016 and May 2017 were evaluated. Pre- and intraoperative recordings were evaluated by three experienced laryngologists. Stroboscopic video images taken during the preoperative diagnosis and interoperative video recordings made before and after AIR and TI were performed were reviewed and compared. During the preoperative evaluation, the surgeons declared their surgical plans and noted changes while observing the intraoperative evaluation during AIR and TI.

Results

Sixty-eight vocal folds were evaluated. The initial diagnosis was found to be consistent with the final diagnosis in only 10 patients (29.4%). The diagnoses of 29 vocal folds (42.6%) and the surgical plans changed after AIR and TI. In six cases, submucosal bands, additional morphological structures in the vicinity of the primary pathology, were observed; these could only be visualized with AIR and TI. AIR and TI revealed new pathologies in four vocal folds that were noted to be normal in the preoperative evaluation.

Conclusion

AIR and TI are useful and promising techniques to identify undiagnosed lesions in vocal folds and to increase the success of minimally invasive phonosurgery.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
The Prevalence of Dysphonia and Dysphagia in Patients with Vitamin D Deficiency

Abdul-Latif Hamdan*
, Elie Khalifee*
, Nader Al Souky*
, Bakr Saridar*
, Pierre Richard Abi Akl*
, Anthony Ghanem*
, Sami Azar†,low asterisklow asterisk,'Correspondence information about the author Sami AzarEmail the author Sami Azar

Journal of Voice

Accepted: March 14, 2019; Published online: April 12, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.007
Publication stage: In Press Corrected Proof

Abstract

Objective

To investigate the prevalence of phonatory and swallowing symptoms in patients with hypovitaminosis D.

Methods/Design

All patients presenting to the endocrinology clinic and investigated for vitamin D deficiency between January 2018 and April 2018 were asked to participate in this study. Demographic data included age, gender, allergy, and history of smoking. Patients filled Voice handicap Index (VHI-10) and Eating Assessment Tool (EAT-10).

Results

A total of 136 consecutive subjects presenting to the endocrinology clinic for vitamin D testing were included: 60 with hypovitaminosis D and 76 with no hypovitaminosis D. The mean vitamin D level in the study group and controls was 13.25 ng/mL and 31.91 ng/mL, respectively. There was no significant difference in the mean score of VHI-10, nor in the mean score of EAT-10 in patients with hypovitaminosis D versus those with no hypovitaminosis D (P value >0.05).

Conclusion

There was no significant difference in the prevalence of phonatory and dysphagia symptoms using VHI-10 and EAT-10 questionnaires between subjects with hypovitaminosis D and those with normal serum vitamin D levels.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Strap Muscle Type I Thyroplasty After Gore‐Tex Implant Extrusion: Case Report and Literature Review

Rishabh Sethia*,†
, Ryan J. Ivancic
, Winslo K. Idicula
, Brad W. deSilva*,†,1,'Correspondence information about the author Brad W. deSilvaEmail the author Brad W. deSilva

Journal of Voice

Accepted: March 19, 2019; Published online: April 10, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.008
Publication stage: In Press Corrected Proof

ABSTRACT

Objectives

To discuss the presentation and management techniques of implant extrusion following type I thyroplasty and to illustrate the potential of strap muscle for augmentation following implant removal.

Methods

We report a unique case of a patient with late Gore-Tex implant extrusion after type I thyroplasty treated with removal and autologous strap muscle graft for augmentation.

Results

A 41-year-old female nearly 3.5 years status post Gore-Tex type I thyroplasty for left vocal fold paralysis presented for evaluation of dysphonia. Upon flexible laryngoscopy, erythema, edema, and granulation tissue were identified at the left vocal fold and ventricle. The patient subsequently underwent removal of her implant. Intraoperatively, a free portion of sternothyroid muscle was dissected free and placed into the paraglottic space. One month following surgery, the patient reported an improvement in her Voice Handicap Index (VHI) score from 40 to 0. In addition, no major complications were observed and complete glottic closure was achieved. Nine months postsurgery, she continued to function well with a VHI score of 0. At 50 months postop, the patient still reports a VHI score of 0.

Conclusions

Implant extrusion is a rare complication of type I thyroplasty usually occurring in the first few months after surgery and more commonly presenting in females. Current management options consist of observation or augmentation with autologous fat or vocal fold injection following implant removal. This is the first report of a successful strap muscle free graft revision thyroplasty following implant extrusion. The patient’s excellent long-term outcome highlights the potential of strap muscle augmentation as a feasible management option for implant extrusion.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Group Voice Therapy Reduces Anxiety in Patients With Dysphonia

Flávia Maiele Pedroza Trajano*,†
, Larissa Nadjara Alves Almeida
, Sauana Alves Leite de Alencar
, João Euclides Fernandes Braga
, Anna Alice Almeida§,low asterisklow asterisk,'Correspondence information about the author Anna Alice AlmeidaEmail the author Anna Alice Almeida

Journal of Voice

Accepted: March 12, 2019; Published online: April 09, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.003
Publication stage: In Press Corrected Proof

Summary

Objective

To evaluate the impact of group therapy in patients with dysphonia, as well as to verify the correlation between vocal symptoms and levels of anxiety.

Methods

The study was composed of 52 patients subdivided into two groups, named the Experimental Group (EG) with 28 volunteers and the Control Group (CG) with 24 volunteers. Anxiety and voice protocols were used for data collection. The State-Trait Anxiety Inventory (STAI) was used to measure trait levels of anxiety that after collection were categorized according to the variation in scores value: low anxiety (20-40 points); average anxiety (40-60 points); high anxiety (60-80 points). In addition, the Voice Symptom Scale (VoiSS) was used for voice assessment. Inferential statistical analysis from the Student’s t test for paired and independent data, in order to compare the average scores of STAI trait levels and VoiSS domains of the pre- and postmoments, intra- and intergroups, EG and CG, respectively. For that purpose, the program Statistical Package for Social Sciences (SPSS) was used.

Results

Half of the patients in the survey presented an average trait level of anxiety. Regarding the EG, there was a significant reduction of state anxiety when comparing the moments before and after group therapy. There was also a significant reduction in the values of the VoiSS-Total and VoiSS-Physical domains when compared to the pre- and postgroup therapy moments. It was verified the existence of a positivecorrelation between the levels of anxiety after group therapy and VoiSS-Total, VoiSS-Limitation, and VoiSS-Physical domains. As for the CG, there was an increase in anxiety levels as well as in all domains of the VoiSS scale when compared to the pre- and postmoments.

Conclusions

Group voice therapy was effective for a significant reduction of vocal symptoms and anxiety – common conditions in patients with dysphonia. It was possible to perceive the positive correlation between anxiety levels and vocal symptoms.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Is There a Relationship Between Vocal Effort and VHI?

Brienne Ruel*,low asterisklow asterisk,'Correspondence information about the author Brienne RuelEmail the author Brienne Ruel
, Susan Thibeault

Journal of Voice

Accepted: March 14, 2019; Published online: April 08, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.005
Publication stage: In Press Corrected Proof

Summary

Objectives

Vocal effort is often a symptom reported when a patient experiences a voice disorder. The aim of this investigation was to compare Voice Handicap Index (VHI) scores among patients with primary complaints of vocal effort (VE) versus vocal quality (VQ) and both vocal effort and quality (VQ+VE). A secondary aim was to compare Glottal Function Index (GFI), and glottic closure pattern between patients with the aforementioned primary presenting complaints and laryngeal pathology diagnosis.

Study Design

Prospective outcomes database design

Methods

Patients were identified with presenting complaints of VQ, VE, or VQ+VE who completed the VHI. Seven hundred and thirteen subjects who met criteria were categorized across three complaint groups and five laryngeal pathologies. Variables selected included age, gender, diagnosis, VHI, GFI, and 12-Item Short Form Health Survey. VHI and GFI scores were compared between the three presenting complaints. Relationships between the categorical variables and glottic closure were also assessed.

Results

Significant differences in total VHI were measured among all three presenting-complaint categories (P < 0.0001). Patients with VE only complaints had significantly greater VHI scores than those patients who report VQ only complaints. VHI scores were significantly higher for patients with combined complaints of VQ+VE as compared to VQ only or VE only. Significant differences in GFI were measured across all three complaints, and GFI was significantly higher in VE only as compared to VQ only (P < 0.001). Glottic closure pattern was not statistically significant in the variables examined. VHI scores were significantly different within pathology subgroups (P < 0.03).

Conclusion

VE only complaints affect quality of life with and without concomitant VQ complaints. Patients with VE complaints perceive a higher level of glottic dysfunction. An underlying mechanism for increased VE could be altered glottic function, however, we were unable to suggest a correlation between glottic closure and GFI.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Prevalence, Characteristics and Impact of Dysphonia in US Marine Corps Drill Instructors

Joseph Spellman*,†
, Michael Coulter*
, Carole Roth*
, Christopher Johnson*,#,'Correspondence information about the author Christopher JohnsonEmail the author Christopher Johnson

Journal of Voice

Accepted: February 27, 2019; Published online: March 26, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.02.015
Publication stage: In Press Corrected Proof

Abstract

Importance

Prior studies have evaluated various populations at increased risk of voice impairment. However, minimal data is available for military Drill Instructors, a population known to have significant vocal demands.

Objective

The purpose of this study was to determine the prevalence of subjective, perceptual, and objective dysphonia in this population and to evaluate contributing factors and impact on job performance.

Design

Cross-sectional analysis.

Setting

United States Marine Corps base (Marine Corps Recruit Depot, San Diego, California).

Participants

Active US Marine Corps Drill Instructors.

Interventions

A survey was administered investigating subjective measures of dysphonia and its impact on occupational function. Standardized voice samples were recorded for objective and perceptual voice analysis.

Main Outcomes and Measures

Prevalence of subjective (Voice Handicap Index-10 and Glottal Function Index scores), perceptual (CAPE-V score), and acoustic (Cepstral-spectral index of dysphonia) measures of dysphonia.

Results

Subjective dysphonia was present in 47.7% by the Voice Handicap Index-10 and 70.2% by the Glottal Function Index. 51% of subjects reported periods of aphonia, while 47% reported voice problems limiting job function in the month prior to being surveyed. The Cepstral-Spectral Index of Dysphonia Mean was abnormal in 95.3% and CAPE-V overall severity score was abnormal in 94%. There was significant improvement in subjective, perceptual, and acoustic voice outcomes as the amount of time since last training cycle (ie, relative voice rest) and as experience as a Drill Instructor increased, however the VHI-10 was the only measure that normalized.

Conclusion and Relevance

There is a very high prevalence of self-reported dysphonia in Drill Instructors, with near-universal prevalence of some degree of objectively and perceptually-rated dysphonia. Nearly half of those surveyed reported that dysphonia limited their job performance. Relative voice rest and experience seem to mitigate severity, but normal ratings were rare. While objective and perceptually-rated dysphonia are persistent and highly prevalent, it does not necessarily translate into a perceived impairment in this population. For these reasons and considering the importance of extraordinary vocal function in this occupation, Drill Instructors appear to be in dire need of proper voice care to both maximize job performance and mitigate long-term voice-related problems.

Published by Elsevier Inc.

Article in Press
Adaptation and Validation of Vocal Fatigue Index (VFI) to Malayalam Language

Usha Rajan Athira
, Usha Devadaslow asterisk,'Correspondence information about the author Usha DevadasEmail the author Usha Devadas
Department of Speech and Hearing, School of Allied Health Sciences, Manipal Academy of Higher Education, Manipal, India

Journal of Voice

Accepted: March 7, 2019; Published online: March 26, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.03.002
Publication stage: In Press Corrected Proof

Abstract

Objective

The purpose of the present study was to translate and validate the English version of the Vocal Fatigue Index (VFI) in to Malayalam language.

Methods

The English version of the VFI was translated into Malayalam language using parallel back translation. The translated version was content validated by three qualified Speech Language Pathologists. The final Malayalam version of the VFI was administered on 528 primary school teachers with and without self-reporting of voice problems. The internal consistency and test-retest reliability were determined using Cronbach’s alpha and intraclass correlation coefficients. Independent sample t test was used to assess the difference in means. Receiver operating characteristic curves with sensitivity and specificity were calculated to set cut-off scores for three domains of Malayalam VFI.

Results

The results revealed that the Malayalam version of VFI has an excellent internal consistency across all the three factors of VFI; tiredness of voice and avoidance of voice use (α = 0.922), physical discomfort symptoms (α = 0.923), and improvement of symptoms with voice rest (α = 0.925). Independent sample t test revealed a significant difference (P ≤ 0.001) for total scores of three domains [tiredness and avoidance of voice use (factor 1), physical discomfort symptoms (factor 2), improvement in symptoms with voice rest (factor 3)] between teachers reporting voice problems and teachers not reporting voice problems. Cut-off scores for three different domains of the Malayalam VFI were set at ≥ 16.5 (80% sensitivity and 71.4% specificity) for factor 1, ≥ 6.5 (71.1% sensitivity and 70.1% specificity) for factor 2 and ≤ 7.5 (71.1% sensitivity and 69.9% specificity) for factor 3.

Conclusion

The Malayalam VFI was found to have good internal consistency and reliability. Hence, it can be considered as a valid and reliable tool for identifying vocal fatigue symptoms and its severity among the Malayalam speaking community who report voice disorders.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Perceptual Judgment of Voice Quality in Nondysphonic French Speakers: Effect of Task-, Speaker- and Listener-Related Variables

Véronique Delvaux*,low asterisklow asterisk,'Correspondence information about the author Véronique DelvauxEmail the author Véronique Delvaux
, Claire Pillot-Loiseau

Journal of Voice

Accepted: February 25, 2019; Published online: March 25, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.02.013
Publication stage: In Press Corrected Proof

Abstract

Purpose

Several perceptual scales have been developed to assess voice quality in dysphonic voices, among which the Grade Roughness Breathiness Asthenia Strain and a Rate of Dysphonia scale is probably the most frequently used. However, this clinical tool has not been properly validated with a normophonic population yet. The aim of the present study was to provide a first set of reference data gathered from a normal population, to serve as a basis of comparison for vocologists and laryngologists working with French-speaking patients. A second goal was to investigate the influence on this normal voice dataset, of variables known to affect perceptual judgments of pathological voice.

Material and methods

Sustained vowels and sentences produced by 80 healthy, normophonic French native speakers were perceptually assessed by a panel of 18 raters (nine students, nine experts) using the Grade Roughness Breathiness Asthenia Strain and a Rate of Dysphonia scale.

Results

The average overall grade was close to 1 on the (0 to 3) scale, questioning the notion of “normal” voice as opposed to dysphonic voice. Rating reliability as well as perceptual scores were affected by task-, speaker-, and listener-related factors: speech stimuli led to better rating reliability and were judged less severely than voice stimuli; experts were slightly more reliable and less severe than students; older speakers were unanimously considered as more dysphonic. Multiple interactions between these factors were observed, confirming the multidimensional nature of voice quality.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Article in Press
Detection of Muscle Tension Dysphonia Using Eulerian Video Magnification: A Pilot Study

Jason Adleberg*
, Ashley P. O’Connell Ferster
, Daniel A. Benito
, Robert T. Sataloff§,low asterisklow asterisk,'Correspondence information about the author Robert T. SataloffEmail the author Robert T. Sataloff

Journal of Voice

Accepted: February 13, 2019; Published online: March 25, 2019
DOI: https://doi.org/10.1016/j.jvoice.2019.02.006
Publication stage: In Press Corrected Proof

Summary

Objective

To determine whether Eulerian Video Magnification software is useful in diagnosis of muscle tension dysphonia (MTD).

Study Design

Prospective.

Methods

Adult patients scheduled in a tertiary care laryngology practice for evaluation of dysphonia were recruited between November 2016 and March 2017. Demographic and clinical data were extracted from patient charts. Diagnosis of MTD was confirmed with videostroboscopic and physical exam and by a speech-language pathologist. Eighteen MTD patients were video recorded while at rest and with phonation. Five patients without MTD also were analyzed as controls. Videos were analyzed using Eulerian Video Magnification software (Massachusetts Institute of Technology) to assess change in blood flow at the forehead, infrahyoid muscles, and sternocleidomastoid muscles, while using the values of the background wall as a control value.

Results

Patients with MTD demonstrated little change in perfusion to the infrahyoid muscles of the neck while phonating (+1% ± 55%). Control subjects demonstrated an increase in perfusion to the infrahyoid muscles while phonating (+102% ± 164%), with this change being significant when comparing the two groups (P = 0.04, t = 2.189, df = 21). A change in perfusion of 0% or less to infrahyoid muscles was 75% sensitive and 70% specific for diagnosis of MTD. No differences in perfusion were found between other regions assessed. Patient age and gender did not correlate with any change in perfusion between rest and phonation.

Conclusion

Our data suggest that Eulerian Video Magnification can be used in the diagnosis of MTD by focusing on the difference in perfusion to the infrahyoid muscles between rest and phonation.

© 2019 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

Voice

Ethics

Three Levels of Ethical Influences on Selling Behavior and Performance: Synergies and Tensions

Abstract

In general, the business ethics literature has treated the conceptual domains and outcomes of macro-level (industrial), meso-level (organizational), and micro-level (individual) ethical influence separately. However, this singular treatment ignores the synergies and tensions that can arise across these different types of ethical influence. Using sales as a research context, the current study argues that all three ethical frames of references are important in shaping employee behavior and performance and, as such, should be examined simultaneously. The findings show that industrial ethical climate and salesperson moral equity are positively associated with salesperson customer orientation. In addition, industrial and organizational ethical norms have a stronger joint effect on customer orientation than either ethical climate alone. More specifically, a more ethical organizational climate enhances the positive effects of the industrial ethical climate on customer orientation. Furthermore, whereas salesperson moral equity is significantly associated with salesperson customer orientation, strong moral equity beliefs in situations requiring adaptive selling result in weaker sales outcomes. This study concludes with a set of theoretical and actionable implications, as well as a discussion of future research avenues.

Going It Alone Won’t Work! The Relational Imperative for Social Innovation in Social Enterprises

Abstract

Shifts in the philosophy of the “state” and a growing emphasis on the “Big Society” have placed an increasing onus on a newly emerging organizational form, social enterprises, to deliver innovative solutions to ease societal issues. However, the question of how social enterprises manage the process of social innovation remains largely unexplored. Based on insights from both in-depth interviews and a quantitative empirical study of social enterprises, this research examines the role of stakeholder relationships in supporting the process of social innovation within social enterprises. We find that social enterprises are adept at working with their stakeholders in the ideation stage of social innovation. In contrast, they often fail to harness knowledge and expertise from their partners during the social innovation implementation phase. Consequently, we propose a social innovation–stakeholder relationship matrix that provides social enterprises in particular with insight for developing stakeholder relationships to achieve their social innovation missions.

Actively Persuading Consumers to Enact Ethical Behaviors in Retailing: The Influence of Relational Benefits and Corporate Associates

Abstract

While consumer motivation to maintain a relationship with a retailer is a function of personal idiosyncratic characteristics, specific perceptions of retailers may play a role in influencing receptivity to relationship maintenance. This study integrates relationship marketing tactics and corporate associates into a model of consumer ethical purchasing behavior that improves the relationship between sellers and buyers. Results show social benefits, special treatment benefits, CSR, and service quality have direct and indirect impact on ethically questionable consumer behaviors in retailing. This study also modifies the consumer ethics scale of Muncy and Vitell (1992) for the East Asian market with good reliability and validity in order to measure ethically questionable consumption behaviors in retailing. Finally, some theoretical contribution and practical implications are discussed.

Who’s Watching? Accountability in Different Audit Regimes and the Effects on Auditors’ Professional Skepticism

Abstract

The European Commission has suggested that the use of joint audits should lead to improved auditor skepticism and—by extension—audit quality, through increased accountability. However, archival research does not find support for improved audit quality in a joint audit setting. To better understand the relationship between accountability in different review regimes and auditors’ judgments, we examine the behavioral effect of implementing a joint audit relative to other review regimes based on a 1 × 3 experimental design. Forty-seven senior auditors and partners from a Big Four firm performed a going concern evaluation task under one of three review regimes: the joint audit, the internal review, and the no review regime. Notwithstanding the difference in the audiences to which auditors are accountable, there is no difference in the judgment process. In terms of their judgment outcome, however, auditors in the joint audit setting were the least skeptical in their judgment of the going concern assumption. Overall, we suggest that the joint audit may lead to unintended behavioral consequences.

Internally Reporting Risk in Financial Services: An Empirical Analysis

Abstract

The enduring failure of financial institutions to identify and deal with risk events continues to have serious repercussions, whether in the form of small but significant losses or major and potentially far-reaching scandals. Using a mixed-methods approach that combines an innovative version of the classic dictator game to inform prosocial tendencies with the survey-based Theory of Planned Behaviour, we examine the risk-escalation behaviour of individuals within a large financial institution. We discover evidence of purely selfish behaviour that explains the lack significance in pressure to adhere to the Subjective Norms of colleagues around intention to report risks. A finding that has potentially important implications for efforts to instil a high-error management climate and incentivise risk reporting within organisations where risk, if ignored or unchecked, could ultimately have consequences that extend far beyond the institutions themselves.

Can Social Norm Activation Improve Audit Quality? Evidence from an Experimental Audit Market

Abstract

We assert that audit quality can be improved to the extent that social norms for honesty and responsibility are activated in the auditor. To test this assertion, we use an experimental audit market setting found in the literature and manipulate factors expected to activate honesty and responsibility norms in the auditor. We find that auditor misreporting is reduced when the investor is another participant in the experiment rather than computer simulated, and thus, the interests of third-party investors are salient to the auditor. We also find that auditor misreporting is reduced when the auditor is required to sign-off on the audit report, but only when the investor is another participant in the experiment. Consistent with our underlying theory, we find that pre-experimental measures of sensitivity to honesty and responsibility norms help explain the effects of our manipulated variables. Finally, we find that these measures of social norm sensitivity are associated with the moral judgment that auditor misreporting is unethical. Our study helps explain previous anomalous findings in the literature and answers the call in Blay et al. (J Bus Ethics 2017. doi:10.1007/s10551-016-3286-4) for empirical researchers to use social norm theory to develop stronger tests of moral reasoning in the market for auditing services.

Achieving Top Performance While Building Collegiality in Sales: It All Starts with Ethics

Abstract

While previous literature provides evidence of the positive relationship between ethical climate and job satisfaction, the possible mechanisms of this relationship are still underexplored. This study aims to enhance scholars’ and practitioners’ understanding of the ethical climate–job satisfaction relationship by identifying and testing two of the possible mechanisms. More specifically, this study fills an existing research gap by examining social and interpersonal mechanisms, referred to in this study as workplace isolation of colleagues and salesperson’s teamwork, of the ethical climate–job satisfaction relationship. This is vital for the selling profession because job satisfaction is known to drive higher levels of salespeople’s performance. The arguments for such mechanisms are built on the foundations of social/psychological contract theory and ethical climate literature. Empirical testing using a large sample of salespeople shows higher levels of ethical climate to decrease workplace isolation and increase teamwork. Findings support hypothesized model where ethical climate positively relates to job satisfaction as partially mediated by workplace isolation and teamwork. Ethical climate is negatively related to workplace isolation and positively to teamwork. Further, findings indicate negative effect of workplace isolation on teamwork and sales performance. Job satisfaction is found to be key factor in driving performance of salespeople.

Perceived Ethical Leadership Affects Customer Purchasing Intentions Beyond Ethical Marketing in Advertising Due to Moral Identity Self-Congruence Concerns

Abstract

Ethical leadership has so far mainly been featured in the organizational behavior domain and, as such, treated as an intra-organizational phenomenon. The present study seeks to highlight the relevance of ethical leadership for extra-organizational phenomena by combining the organizational behavior perspective on ethical leadership with a classical marketing approach. In particular, we demonstrate that customers may use perceived ethical leadership cues as additional reference points when forming purchasing intentions. In two experimental studies (N = 601 and N = 336), we find that ethical leadership positively affects purchasing intentions because of customers’ concerns for moral self-congruence. We show this by means of both mediation and moderation analyses. Interestingly, the effect of perceived ethical leadership on purchasing intentions holds over and above the ethical advertising claims (e.g., cause-related marketing) that are commonly used in marketing. We conclude by discussing the possible ramifications of ethical leadership beyond its effects on immediate employees.

How and When Retailers’ Sustainability Efforts Translate into Positive Consumer Responses: The Interplay Between Personal and Social Factors

Abstract

This study aims to address how (through which mechanisms) and when (under which conditions) retailers’ sustainability efforts translate into positive consumer responses. Hypotheses are developed and tested through a scenario-based experiment among 672 consumers. Retailers’ assortment sustainability and distribution sustainability are manipulated. Retailers’ sustainability efforts lead to positive consumer responses (e.g., improved store evaluations) via two underlying mechanisms: consumers’ identification with the store (personal route) and store legitimacy (social route). The effects of sustainability efforts are strengthened if consumers have personal norms favoring shopping at environmentally friendly stores. Remarkably, when controlling for moderation by personal norms, social norms weaken the effects. The findings show that traditional marketing mix elements provide opportunities for retailers to improve their organizations’ bottom line and positively affect consumer (and societal) well-being. This study helps retailers decide whether or not to invest in and communicate about sustainability. Past research has shown the clear potential for positive consumer responses to firms’ sustainability efforts, but little is known about the underlying mechanisms and the conditions under which such responses take place. This study advances theory by examining personal and social factors as mediators and moderators of the retailers’ sustainability efforts–consumer responses relationship.

The Role of the Distributor Network in the Persistence of Legal and Ethical Problems of Multi-level Marketing Companies

Abstract

Multi-level marketing companies (MLMs) such as Amway, Herbalife, or Tupperware differ from most other companies. They market their products and services by means of self-employed distributors who typically work from home, sell products to end consumers, and recruit, motivate, and educate new distributors to do the same. Although the industry’s growth seems to illustrate the attractiveness of MLMs, the industry has been facing several legal and ethical problems. In this paper, we focus on these problems and argue that an extended MLM model may help us to understand why such problems continue to occur, despite the countermeasures that have been implemented. By explicating how problems relate to a specific but often overlooked characteristic of MLMs, i.e., the so-called distributor network, we provide an extended understanding of (a) MLMs’ mode of operation, (b) the sources of their legal and ethical problems, and (c) the reason that currently implemented and suggested countermeasures may not suffice. Moreover, based on our extended understanding of MLMs and their problems, we propose additional countermeasures.

Ethics

Der Onkologe

Therapiealgorithmus bei primärem Vaginalkarzinom
Moderne Strahlentherapie des Vulvakarzinoms

Zusammenfassung

Hintergrund

In den letzten 20 Jahren haben sich die onkologischen Ergebnisse in der Behandlung des Vulvakarzioms nicht verbessern lassen. Aufgrund fehlender Evidenzen aus randomisierten Studien sind zahlreiche Aspekte der Behandlung unklar und kontrovers. Therapieziele aus radioonkologischer Sicht sind die Vermeidung lokaler und lokoregionärer Rezidive, die Verbesserung von Organ- und Funktionserhalt im Rahmen des interdisziplinären Therapiekonzepts und die Verhinderung unnötiger Toxizitäten.

Methode

Es erfolgte eine Literaturrecherche und eine systematische Darstellung der aktuellen Therapiestandards der radioonkologischen Behandlung von Vulvakarzinomen.

Ergebnisse

Tumorbefallene Resektionsränder erfordern eine lokale Nachresektion oder, falls nicht möglich oder funktionell ungünstig, eine postoperative Radio(chemo)therapie. Die aktuelle deutsche Leitlinie empfiehlt derzeit eine Resektion von mindestens 3 mm im Gesunden.

Die Vermeidung der inguinalen Rezidive verbessert das krankheitsfreie Überleben. Die häufig geübte Praxis der pelvinen Bestrahlung ist selbst für Patientinnen mit positiven inguinalen Lymphknoten in ca. 70 % eine Übertherapie. Hier hat sich zur Vermeidung unnötiger Toxizitäten eine Neubewertung durchsetzen können. Die aktuelle deutsche Leitlinie empfiehlt ein operatives Staging des Beckens und eine pelvine Bestrahlung nur im Falle positiver Lymphknoten.

Schlussfolgerung

Themen der Diskussion und zukünftiger Neubewertung sind der Stellenwert der simultanen Chemotherapie, der neoadjuvanten vs. primären Radiochemotherapie, der optimalen radiosensibilisierenden Substanz und der optimalen Bestrahlungsdosis in der primären Therapie. Die Interdisziplinarität stellt eine unabdingbar Voraussetzung für eine optimale Therapie dar.

Moderne Strahlentherapie des Vaginalkarzinoms

Zusammenfassung

Hintergrund

Das primäre Karzinom der Vagina ist eine sehr seltene Entität. Die Therapiestrategie beim Vaginalkarzinom orientiert sich an den Behandlungskonzepten von Plattenepithelkarzinomen der Zervix und Vulva.

Methode

Es erfolgte eine Literaturrecherche und eine systematische Darstellung der prognostischen Faktoren, der aktuellen Therapiestandards und des klinischen Outcomes der radioonkologischen Behandlung des Vaginalkarzinoms.

Ergebnisse

Die Strahlentherapie ist Therapie der Wahl bei der Behandlung von lokalen Vaginalkarzinomen. Tumorstadium, Lymphknotenstatus und Resektionsstatus sind etablierte prognostische Faktoren. Die Tumorgröße (>4 cm) ist prognostisch v. a. bei Patientinnen mit alleiniger Strahlentherapie relevant. Moderne Bestrahlungstechniken haben die Toxizität der Behandlung stark vermindert. Brachytherapie ist von entscheidender Bedeutung für die Krankheitskontrolle. Seit der Etablierung der Radiochemotherapie (RCT) konnte in den Stadien II–IVa eine deutliche Verbesserung des Therapieergebnisses erreicht werden (Stadium II 41,7 % auf 58,1 %, Stadium III 24 % auf 43 % im Fünfjahresgesamtüberleben).

Schlussfolgerung

Durch eine moderne radioonkologische Behandlung können beim Vaginalkarzinom gute Behandlungsergebnisse mit akzeptablen Nebenwirkungen erreicht werden. Eine präzise Ausbreitungsdiagnostik ermöglicht eine individuelle Behandlungsstrategie und hilft, operative und strahlentherapeutische Über- oder Untertherapien zu vermeiden. Die Behandlung sollte interdisziplinär und in erfahrenen Zentren erfolgen.

Epidemiologie der Krebserkrankungen von Vulva und Vagina in Deutschland

Zusammenfassung

Für die Diagnosejahre seit 2009 ist die bevölkerungsbezogene Krebsregistrierung in ganz Deutschland flächendeckend umgesetzt. Das Zentrum für Krebsregisterdaten führt seitdem jährlich die Krebsregisterdaten der Länder zusammen und schätzt deren Vollzähligkeit. Die bundesweiten Krebsneuerkrankungen von Vulva und Vagina (definiert als C51 bzw. C52 nach der derzeit gültigen 10. Fassung der Internationalen Klassifikation der Krankheiten und verwandter Gesundheitsprobleme, ICD-10) wurden auf der Basis vollständiger Landeskrebsregister geschätzt. Aktuell treten jährlich etwa 3580 Neuerkrankungen auf, wovon knapp 90 % die Vulva betreffen. Eine Zunahme der Krebserkrankungen der Vulva hat in den vergangenen Jahren auch zu einer Zunahme der Sterbefälle geführt. Etwa 930 Frauen verstarben 2016 an Vulvakrebs; weitere 190 Frauen verstarben am Vaginalkarzinom. Im jungen Erwachsenenalter treten bösartige Vulva- und Vaginaltumoren nur sehr selten auf; die Erkrankungsraten steigen mit zunehmendem Lebensalter kontinuierlich an. Betroffene Frauen sind zum Diagnosezeitpunkt einige Jahre älter (medianes Erkrankungsalter 73 Jahre) als Frauen mit anderen gynäkologischen Erkrankungen wie Eierstock- oder Gebärmutterkörperkrebs (medianes Erkrankungsalter 69 Jahre). Eine von 3 Patientinnen mit Krebs der Vulva wird 5 Jahre nach der Diagnosestellung an ihrer Erkrankung verstorben sein (relatives Überleben: 68 %). Bei Frauen mit einer Krebserkrankung der Vagina ist die Prognose schlechter. Das relative Fünfjahresüberleben beträgt 43 %.

Bildgebende Diagnostik bei Karzinomen der Vulva und Vagina

Zusammenfassung

Hintergrund

Eine bildgebende Diagnostik mittels Magnetresonanztomographie (MRT), Computertomographie (CT) oder PET-CT (Positronen-Emissions-Tomographie-CT) wird bei gynäkologischen Tumoren zum Staging, zur Therapieplanung und -kontrolle oder zur Rezidivdiagnostik zunehmend eingesetzt.

Ziel

Es wird der aktuelle Stand des Einsatzes der Schnittbildgebung beim Vulva- und Vaginalkarzinom beleuchtet.

Material und Methoden

Es erfolgte eine Recherche aktueller Literatur und der gültigen deutschen Leitlinien.

Ergebnisse

Die funktionelle MRT des Beckens (T2-Wichtung, Diffusionswichtung, Kontrastmittel-Dynamik) hat ihren Stellenwert beim Vulva- und Vaginalkarzinom im lokalen Staging und bei der Rezidivdiagnostik. Die kontrastmittelverstärkte CT wird zur Fernmetastasensuche eingesetzt. Die PET-CT kann in Einzelfällen zum Einsatz kommen, beispielsweise vor geplanter Exenteration.

Schlussfolgerungen

Die Schnittbildgebung liefert therapierelevante Details und Zusatzinformationen beim Vulva- und Vaginalkarzinom bzw. in der Rezidivsituation.

Aktuelle Studien und experimentelle Ansätze in der Therapie von Karzinomen der Vulva- und Vagina

Zusammenfassung

Hintergrund

Es gibt wenige prospektiv durchgeführte und bis dato abgeschlossene Studien zum Thema Therapie eines Vulva- oder Vaginalkarzinoms.

Ziel

Eine selektive Literaturrecherche in der Datenbank Pubmed sowie der webbasierten Datenbank www.clinicaltrial.gov wurde durchgeführt.

Material und Methodik

Mittels Schlüsselwörter wie „Vulvakarzinom, Vaginalkarzinom, Therapie, Studien“ wurde die Recherche durchgeführt.

Ergebnisse

Eine selektive Übersicht von publizierten Arbeiten wurde zusammengestellt. Es konnten publizierte Arbeiten für das Vulvakarzinom und Vaginalkarzinom gesichtet werden.

Schlussfolgerung

Die Datenlage für die o. g. Tumorentität ist weiterhin sehr dünn. Demnächst sollten neueste Ergebnisse im Einklang mit Kenntnissen der Tumorbiologie veröffentlicht werden.

Prophylaxe des Vulva- und des Vaginalkarzinoms und ihrer Vorstufen

Zusammenfassung

Hintergrund

Vulva- und Vaginalkarzinome sind zunehmend diagnostizierte Tumorentitäten in Deutschland. Vulva- und Vaginalkarzinome entstehen über zwei unterschiedliche pathogenetische Mechanismen: Zum einen kann eine persistierende Infektion mit dem humanen Papillomvirus (HPV, v. a. Typ 16, 31, 33) über eine undifferenzierte vulväre/vaginale intraepitheliale Neoplasie (VIN/VAIN) zum invasiven Vulva‑/Vaginalkarzinom führen. Diese Tumoren finden sich eher bei jüngeren Frauen. Davon abzugrenzen ist das Vulvakarzinom der älteren Frauen, das auf dem Boden einer chronischen Hauterkrankung (z. B. Lichen sclerosus et atrophicus, LSA) über Mutationen im Tumorsuppressorgen p53 entsteht. Auch beim Vaginalkarzinom ist einer der Risikofaktoren ein Lichen sclerosus oder Lichen planus, diese Tumoren sind HPV-negativ. Daneben gibt es eine ganze Reihe von Tumoren, deren Ursache nicht bekannt ist.

Fragestellung

Gibt es primäre oder sekundäre Präventionsmaßnahmen, die die Entstehung dieser Tumoren verhindern können?

Ergebnisse

Die HPV-induzierten Vulva- und Vaginalkarzinome können zu einem bedeutenden Teil durch eine HPV-Impfung von HPV-naiven Frauen primär verhindert werden. Der nonavalente Impfstoff deckt die relevanten HPV-Typen 16, (18), 31 und 33 mit einer Wirksamkeit von 97 % ab, der bivalente Impfstoff hat eine vergleichbar hohe Effektivität aber nur für HPV16 und HPV18.

Ob das licheninduzierte Vulva‑/Vaginalkarzinom durch eine konsequente lokale Dauertherapie mit Kortison oder Calcineurininhibitoren primär präventiv verhindert werden kann wird diskutiert. Erste Daten, die darauf hindeuten, liegen vor.

Karzinome der Vulva und Vagina − ein Update
Partnerschaft und Sexualität bei Tumorerkrankungen

Zusammenfassung

Eine Krebserkrankung stellt nicht nur für den Patienten, sondern auch für die Angehörigen eine belastende Situation dar. Insbesondere die Partner erleben häufig vergleichbare psychische und soziale Belastungen wie die Patienten. Dabei spielen die Partner eine wichtige Rolle in der emotionalen und praktischen Unterstützung des Patienten. Die Krebserkrankung kann zu massiven Änderungen der partnerschaftlichen Funktionsfähigkeit wie z. B. Rollenveränderungen oder Einschränkung gemeinsamer Aktivitäten sowie sexuellen Funktionsstörungen führen, die Anpassungsprozesse erforderlich machen. Sexuelle Themen führen oft zu einer Sprachlosigkeit zwischen Patient und Partner, aber auch zwischen dem Paar und dem medizinischen Behandlungsteam. Eine offene Kommunikation über sexuelle Themen – im Idealfall initiiert vom medizinischen Team – wäre empfehlenswert. In der Partnerschaft haben sich ebenfalls eine offene Kommunikation sowie die Förderung des dyadischen Copings (der gemeinsamen Bewältigung des Paares) als hilfreich für die Belastungsreduktion und Steigerung der Lebensqualität erwiesen. Partnerschaftliche Unterstützungsprogramme sind dabei eine sinnvolle Hilfe für Paare in diesem Anpassungs- und Bewältigungsprozess.

Gemcitabin und Oxaliplatin bei Karzinomen der Gallenwege
Der Onkologe

Genetics

Correction to: Splicing mutations in human genetic disorders: examples, detection, and confirmation
The original version on this paper contained an error. The first names and last names of Anna Abramowicz and Monika Gos are inadvertently interchanged and are incorrectly displayed in indexing sites. The correct names are presented above.
Co-treatment with probucol does not improve lung pathology in hydroxypropyl-β-cyclodextrin-treated Npc1 −/− mice

Abstract

We previously reported the altered pulmonary function and pathology found in the mouse model of infantile Niemann-Pick C1 disease, the Npc1−/− mouse. Despite its salutary properties on brain and liver parameters, we did not find efficacious effects of hydroxypropyl-β-cyclodextrin (HPBCD) on pulmonary pathology. Since we had previously shown the beneficial effects of probucol on the somatic phenotype in the Npc1−/− mice, we have now studied the effects of combined therapy with HPBCD and probucol on the lung with mostly negative results. Body weight and lung weight for body weight were increased in parallel while inspiratory capacity for body weight was markedly decreased. Other physical, biochemical, and pulmonary function parameters were not much changed. There were trends towards improved lung elastance (p = 0.09) and compliance (p = 0.07).

Zebrafish breeding program: genetic parameters estimates for growth traits

Abstract

The objective of this study was to evaluate the genetic parameters of two generations of zebrafish breeding program. The base population was formed by crossing individuals of six commercial stocks of zebrafish, resulting in a nucleus with 60 families. Two generations were evaluated, with a total of 780 and 781 individuals for the first and second generation, respectively. The selection was made based on the mean genetic value of each family, followed by mass selection of the breeders. Mathematical models that considered the fixed (age, density in the larval stage, sex, and generation) and random (animal additive genetics, common to full-sibs, and residual) effects were evaluated using BLUPF90 program family. Weight and total length were used as response variables. Total length was the best selection criterion because it had a higher heritability (0.30) than weight (0.22). There was a high common to full-sib effect, especially in the first generation of animals. For second-generation data, the heritability was 0.26 for total length, as well as a lower common to full-sib effect for length. The best model obtained for this evaluation was considering all effects, being age and density as first and second polynomial, respectively. The genetic and phenotypic correlations for weight and length were 0.87 and 0.75, respectively. These results indicate that genetic breeding using total length as the selection criterion may produce a larger and heavier zebrafish strain.

New QTL for resistance to Puccinia polysora Underw in maize

Abstract

Southern corn rust (SCR) is a prevalent foliar disease in maize. Deployment of resistant cultivars is an effective way to control SCR. In this study, resistance to SCR was evaluated in a BC1RIL population comprising 118 lines grown under three different field conditions. Combined with a genetic map constructed from 1635 SNP markers obtained from the maize 9.4 K SNP Affymetrix® Axiom® Genotyping Array, single quantitative trait loci (QTL) were mapped on chromosomes 4, 9, and 10, respectively. The QTL on chromosome 4 (qSCR4.08) and chromosome 9 (qSCR9.04) were stable across multiple environments, and each explained more than 10% of the phenotypic variation. The stable QTL detected could be desirable sources of SCR resistance in maize-breeding programs.

Genotype by environment interaction using AMMI model and estimation of additive and epistasis gene effects for 1000-kernel weight in spring barley ( Hordeum vulgare L.)

Abstract

The objective of this study was to assess genotype by environment interaction for 1000-kernel weight in spring barley lines grown in South Poland by the additive main effects and multiplicative interaction model. The study comprised of 32 spring barley (Hordeum vulgare L.) genotypes (two parental genotypes—breeding line 1 N86 and doubled haploid (DH) line RK63/1, and 30 DH lines derived from F1 hybrids), evaluated at six locations in a randomized complete block design, with three replicates. 1000-kernel weight ranged from 24.35 g (for R63N/42 in 2011) to 61.46 g (for R63N/18 in 2008), with an average of 44.80 g. AMMI analyses revealed significant genotype and environmental effects as well as GE interaction with respect to 1000-kernel weight. In the analysis of variance, 16.86% of the total 1000-kernel weight variation was explained by environment, 32.18% by differences between genotypes, and 24.50% by GE interaction. The lines R63N/61, R63N/22, and R63N/1 are recommended for further inclusion in the breeding program because their stability and the highest averages of 1000-kernel weight. The total additive effect of all genes controlling the trait and the total epistasis effect of 1000-kernel weight were estimated. Additive gene action effects based on DH lines were always larger that this parameter estimated on the basis of parental lines. Estimates of additive gene action effects based on the all DH lines were significantly larger than zero in each year of study. Epistasis effects based on all DH lines were statistically significant in 2011 and 2013.

Comparing assignment-based approaches to breed identification within a large set of horses

Abstract

Considering the extensive data sets and statistical techniques, animal breeding embodies a branch of machine learning that has a constantly increasing impact on breeding. In our study, information regarding the potential of machine learning and data mining within a large set of horses and breeds is presented. The individual assignment methods and factors influencing the success rate of the procedure are compared at the Czech population scale. The fixation index values ranged from 0.057 (HMS1) to 0.144 (HTG6), and the overall genetic differentiation amounted to 8.9% among the breeds. The highest genetic divergence (FST = 0.378) was established between the Friesian and Equus przewalskii; the highest degree of gene migration was obtained between the Czech and Bavarian Warmblood (Nm = 14,302); and the overall global heterozygote deficit across the populations was 10.4%. The eight standard methods (Bayesian, frequency, and distance) using GeneClass software and almost all mainstream classification algorithms (Bayes Net, Naive Bayes, IB1, IB5, KStar, JRip, J48, Random Forest, Random Tree, PART, MLP, and SVM) from the WEKA machine learning workbench were compared by utilizing 314,874 real allelic data sets. The Bayesian method (GeneClass, 89.9%) and Bayesian network algorithm (WEKA, 84.8%) outperformed the other techniques. The breed genomic prediction accuracy reached the highest value in the cold-blooded horses. The overall proportion of individuals correctly assigned to a population depended mainly on the breed number and genetic divergence. These statistical tools could be used to assess breed traceability systems, and they exhibit the potential to assist managers in decision-making as regards breeding and registration.

A novel WDR62 missense mutation in microcephaly with abnormal cortical architecture and review of the literature

Abstract

Autosomal recessive primary microcephaly (MCPH) is a group of rare neurodevelopmental diseases with severe microcephaly at birth. One type of the disorder, MCPH2, is caused by biallelic mutations in the WDR62 gene, which encodes the WD repeat–containing protein 62. Patients with WDR62 mutation may have a wide range of malformations of cortical development in addition to congenital microcephaly. We describe two patients, a boy and a girl, with severe congenital microcephaly, global developmental delay, epilepsy, and failure to thrive. MRI showed hemispherical asymmetry, diffuse pachygyria, thick gray matter, indistinct gray-white matter junction, and corpus callosum and white matter hypoplasia. Whole exome sequencing revealed the same novel homozygous missense mutation, c.668T>C, p.Phe223Ser in exon 6 of the WDR62 gene. The healthy parents were heterozygous for this mutation. The mutation affects a highly conserved region in one of the WD repeats of the WDR62 protein. Haplotype analysis showed genetic relatedness between the families of the patients. Our findings expand the spectrum of mutations randomly distributed in the WDR62 gene. A review is also provided of the brain malformations described in WDR62 mutations in association with congenital microcephaly.

A single enzyme PCR-RFLP assay targeting V1-V3 region of 16S rRNA gene for direct identification of Alicyclobacillus acidoterrestris from other Alicyclobacillus species

Abstract

Alicyclobacillus acidoterrestris is considered to be one of the most important target microorganisms in quality control of heat-processed acidic foods. This species can cause spoilage by forming spores with very high heat resistance; however, no gas production or visible changes occur in the contaminated product during storage. Thus, the differentiation of A. acidoterrestris from other Alicyclobacillus species is of great importance. Τhe present study aims to find a rapid method for the identification of A. acidoterrestris. To achieve this, 78 Alicyclobacillus isolates were subjected to PCR-RFLP (restriction fragment length polymorphism). Specifically, PCR products of amplified V1-V3 region of the 16S rRNA gene were digested with restriction endonuclease HhaI. According to the obtained results, all A. acidoterrestris isolates showed similar restriction fragments of the 16S rRNA gene and different from A. acidocaldarius and A. hesperidum. In conclusion, a single enzyme PCR-RFLP assay was developed and showed rapid, inexpensive and direct identification of Alicyclobacillus isolates. The application of this method will be useful to identify this contaminant in fruit juices.

Novel data on growth phenotype and causative genotypes in 29 patients with Morquio (Morquio-Brailsford) syndrome from Central-Eastern Europe

Abstract

Mucopolysaccharidosis type IVA, also known as Morquio (Morquio-Brailsford) syndrome results from accumulation of keratan sulfate (KS) and chondroitin-6-sulfate (C6S), whereas the primary cause is mutations in the gene encoding galactosamine (N-acetyl)-6-sulfatase (GALNS). Phenotypically it seems to be a well-defined condition, with two main clinical forms: mild (attenuated) and severe, which are determined based on a combination of symptoms, i.e., enzymatic activity of GALNS, age of onset, and symptom severity. Nevertheless, the natural history of MPSIVA in relation to specific anthropometric parameters (growth, head circumference, body proportions, and face phenotype) is not precisely characterized. The aim of our work was to analyze the aforementioned anthropometric parameters, including correlation to molecular data (causative GALNS mutations).

Copy number variation of bovine SHH gene is associated with body conformation traits in Chinese beef cattle

Abstract

Sonic Hedgehog (Shh) regulates many key developmental processes during vertebrate limb development, fat formation, and skeletal tissue regeneration. Current whole genome sequencing data have identified a copy number variation mapping to bovine Sonic Hedgehog gene (SHH-CNV). The object of this study was to characterize the SHH-CNV distributions in 648 individuals from 11 Chinese cattle populations and further to investigate the associations of the copy number changes with gene expression and cattle growth traits. The SHH-CNV showed a high variance within Chinese indigenous yellow cattle. Compared to yak and dairy cattle, the beef cattle like Luxi and Xianan breed had significantly higher median copy numbers, suggesting the diversity of SHH-CNV in beef cattle selections. The negative correlation of SHH-CNV with SHH transcriptional level in adult adipose tissue (P < 0.01) indicated the dosage effects of SHH-CNV related to bovine fat formation. Association analysis of SHH-CNV and body size traits was conducted in five breeds. The results revealed that the copy number gain type of SHH-CNV exhibited significantly better chest depth in 24 months old Qinchuan cattle, and better body weight, body length, and chest girth in 18 months old Nanyang cattle, whereas the normal copy number had superior chest girth and body weight in adult Jinnan cattle (P < 0.05 or P < 0.01). In summary, this research uncovered meaningful effects of SHH-CNV on gene expression and cattle phenotypic traits, indicating its potential applications for genetic improvement of beef cattle.

Genetics