Bi-allelic variants in DNA mismatch repair proteins MutS Homolog MSH4 and MSH5 cause infertility in both sexes.

STUDY QUESTION: Do bi-allelic variants in the genes encoding the MSH4/MSH5 heterodimer cause male infertility? SUMMARY ANSWER: We detected biallelic, (likely) pathogenic variants in MSH5 (4 men) and MSH4 (3 men) in six azoospermic men, demonstrating that genetic variants in these genes are a relevant cause of male infertility. WHAT IS KNOWN ALREADY: MSH4 and MSH5 form a heterodimer, which is required for prophase of meiosis I. One variant in MSH5 and two variants in MSH4 have been described as causal for premature ovarian insufficiency (POI) in a total of five women, resulting in infertility. Recently, pathogenic variants in MSH4 have been reported in infertile men. So far, no pathogenic variants in MSH5 had been described in males. STUDY DESIGN, SIZE, DURATION: We utilized exome data from 1305 men included in the Male Reproductive Genomics (MERGE) study, including 90 males with meiotic arrest (MeiA). Independently, exome sequencing was performed in a man with MeiA from a large consanguineous family. PARTICIPANTS/MATERIALS, SETTING, METHODS: Assuming an autosomal-recessive mode of inheritance, we screened the exome data for rare, biallelic coding variants in MSH4 and MSH5. If possible, segregation analysis in the patients' families was performed. The functional consequences of identified loss-of-function (LoF) variants in MSH5 were studied using heterologous expression of the MSH5 protein in HEK293T cells. The point of arrest during meiosis was determined by γH2AX staining. MAIN RESULTS AND THE ROLE OF CHANCE: We report for the first time (likely) pathogenic, homozygous variants in MSH5 causing infertility in 2 out of 90 men with MeiA and overall in 4 out of 902 azoospermic men. Additionally, we detected biallelic variants in MSH4 in two men with MeiA and in the sister of one proband with POI. γH2AX staining revealed an arrest in early prophase of meiosis I in individuals with pathogenic MSH4 or MSH5 variants. Heterologous in vitro expression of the detected LoF variants in MSH5 showed that the variant p.(Ala620GlnTer9) resulted in MSH5 protein truncation and the variant p.(Ser26GlnfsTer42) resulted in a complete loss of MSH5. LARGE SCALE DATA: All variants have been submitted to ClinVar (SCV001468891-SCV001468896 and SCV001591030) and can also be accessed in the Male Fertility Gene Atlas (MFGA). LIMITATIONS, REASONS FOR CAUTION: By selecting for variants in MSH4 and MSH5, we were able to determine the cause of infertility in six men and one woman, leaving most of the examined individuals without a causal diagnosis. WIDER IMPLICATIONS OF THE FINDINGS: Our findings have diagnostic value by increasing the number of genes associated with non-obstructive azoospermia with high clinical validity. The analysis of such genes has prognostic consequences for assessing whether men with azoospermia would benefit from a testicular biopsy. We also provide further evidence that MeiA in men and POI in women share the same genetic causes. STUDY FUNDING/COMPETING INTEREST(S): This study was carried out within the frame of the German Research Foundation sponsored Clinical Research Unit 'Male Germ Cells: from Genes to Function' (DFG, CRU326), and supported by institutional funding of the Research Institute Amsterdam Reproduction and Development and funds from the LucaBella Foundation. The authors declare no conflict of interest.

Challenges for Adoption of Integrated Pest Management (IPM): the Soybean Example.

Soybean is considered one of today's most important crops. Planted on millions of hectares worldwide, the management of soybean pests usually requires large amounts of chemicals. However, a key component to meet the increasing demand for food due to the rapidly growing global population is protecting crops from pests while maintaining environmental quality through ecologically and economically sound integrated pest management (IPM) practices. Not only can IPM result in more profitable agriculture due to the reduction of pest control costs but also assures equitable, secure, sufficient, and stable flows of both food and ecosystem services. Despite those ecological and economic benefits, the vast areas of cultivated soybean as well as the convenience of spraying insecticides are encouraging the adoption of prophylactic pest control as a relatively inexpensive safeguard compared to IPM practices. Thus, in this forum, we discuss the reasons for soybean IPM not reaching its potential. We give examples of how we can revive this once successful pest management program with a focus on experiences in Brazil and the USA. We analyze IPM case studies to illustrate the need for growers to have easy and fast access to IPM information on its medium- and long-term benefits. Overall, this forum highlights the importance of IPM for agricultural sustainability including ecological and financial benefits.

Prevalence, risk factors, and type of sleep apnea in patients with paroxysmal atrial fibrillation.

BACKGROUND: Recent studies have suggested an association between sleep apnea (SA) and atrial fibrillation (AF). We aimed to study the prevalence, characteristics, risk factors and type of sleep apnea (SA) in ablation candidates with paroxysmal AF. METHODS/RESULTS: We prospectively studied 579 patients with paroxysmal AF, including 157 women (27.1%) and 422 men (72.9%). Mean age was 59.9 ± 9.6 years and mean body mass index (BMI) 28.5 ± 4.5 kg/m2. SA was diagnosed using polygraphy for two nights at home. The Epworth Sleepiness Scale (ESS), STOP-Bang Questionnaire, and Berlin Questionnaire (BQ) assessed the degree of SA symptoms. A total of 479 (82.7%) patients had an apnea-hypopnea index (AHI) ≥ 5, whereas moderate-severe SA (AHI ≥ 15) was diagnosed in 244 patients (42.1%). The type of SA was predominantly obstructive, with a median AHI of 12.1 (6.7-20.6) (range 0.4-85.8). The median central apnea index was 0.3 (0.1-0.7). AHI increased with age, BMI, waist and neck circumference, body and visceral fat. Using the Atrial Fibrillation Severity Scale and the SF-36, patients with more severe SA had a higher AF burden, severity and symptom score and a lower Physical-Component Summary score. Age, male gender, BMI, duration of AF, and habitual snoring were independent risk factors in multivariate analysis (AHI ≥ 15). We found no association between ESS and AHI (R2 = 0.003, p = 0.367). CONCLUSIONS: In our AF population, SA was highly prevalent and predominantly obstructive. The high prevalence of SA detected in this study may indicate that SA is under-recognized in patients with AF. None of the screening questionnaires predicted SA reliably.

Egg Albumin as a Protein Marker to Study Dispersal of Noctuidae in the Agroecosystem.

Knowledge of dispersal and spatial dynamics of pest populations is fundamental for implementation of integrated pest management and integrated resistance management. This study evaluated 1) the effectiveness of egg white albumin protein to mark larvae and adults of two polyphagous and highly mobile pests, Spodoptera frugiperda (J.E. Smith) (fall armyworm) and Helicoverpa zea (Boddie) (corn earworm) (Lepidoptera: Noctuidae), and 2) the sensitivity of polyvinylidene difluoride membrane (dot blot) in detecting albumin on marked insects. Laboratory and field experiments tested egg albumin as a protein marker, which was detected using two enzyme-linked immunosorbent assay (ELISA), microplate, and dot blot. In the laboratory, 100% of the moths sprayed with 20% egg white solution acquired the albumin marker, which was detected through the last time point tested (5 d) after application. Egg albumin was not effective at long-term marking of larvae, detected only prior the molting to the next instar. Albumin application in field cages resulted in a high percentage of moths detected as marked at 24 h and 5 d for both species. Egg albumin applied in the open field resulted in 15% of the recaptured corn earworm moths marked with most of them collected 150 m from the application area, although some were captured as far as 1,600 m within approximately 6 d after adult emergence. The results indicated egg albumin is a suitable marker to study the dispersion of fall armyworm and corn earworm in the agroecosystem and dot blot was as effective to detect egg albumin as was indirect ELISA.

First Documented Pathologies in Tenontosaurus tilletti with Comments on Infection in Non-Avian Dinosaurs.

In 2001, a nearly complete sub-adult Tenontosaurus tilletti was collected from the Antlers Formation (Aptian-Albian) of southeastern Oklahoma. Beyond its exceptional preservation, computed tomography (CT) and physical examination revealed this specimen has five pathological elements with four of the pathologies a result of trauma. Left pedal phalanx I-1 and left dorsal rib 10 are both fractured with extensive callus formation in the later stages of healing. Left dorsal rib 7 (L7) and right dorsal rib 10 (R10) exhibit impacted fractures compressed 26 mm and 24 mm, respectively. The fracture morphologies in L7 and R10 indicate this animal suffered a strong compressive force coincident with the long axis of the ribs. All three rib pathologies and the pathological left phalanx I-1 are consistent with injuries sustained in a fall. However, it is clear from the healing exhibited by these fractures that this individual survived the fall. In addition to traumatic fractures, left dorsal rib 10 and possibly left phalanx I-1 have a morphology consistent with post-traumatic infection in the form of osteomyelitis. The CT scans of left metacarpal IV revealed the presence of an abscess within the medullary cavity consistent with a subacute form of hematogenous osteomyelitis termed a Brodie abscess. This is only the second reported Brodie abscess in non-avian dinosaurs and the first documented occurrence in herbivorous dinosaurs. The presence of a Brodie abscess, known only in mammalian pathological literature, suggest mammalian descriptors for bone infection may be applicable to non-avian dinosaurs.

Characterization and Identification of Methods for Phenotyping Soybean Populations With Tolerance to the Soybean Aphid (Hemiptera: Aphididae).

The development of soybeans tolerant to the soybean aphid [Aphis glycines Matsumura (Hemiptera: Aphididae)] remains unexplored. The objectives of this research were to determine the susceptibility of two high-yielding soybean [Glycine max (L.) Merrill (Fabales: Fabaceae)] genotypes involved in a breeding platform to develop aphid-tolerant recombinant inbred lines (RILs); characterize the peroxidase activity and relative expression of peroxidase transcripts in the parents of RILs; and identify an assay to phenotype aphid-tolerant RILs. Enzyme kinetic assays documented the total peroxidase activity for tolerant (KS4202), susceptible (SD76R), and two high-yielding (U09-105007 and U11-611112) soybeans during two vegetative stages (V1 and V3) at three sampling days (D4, D6, and D8 after aphid introduction). Enzyme kinetic assays showed that V3 infested tolerant and U11-611112 plants had significantly higher peroxidase activity than their respective control plants at D4, and infested tolerant plants were also higher than control plants at D6. There were no apparent trends when comparing the expression of peroxidase-specific transcripts in the absence of aphids (basal levels) in both V1 and V3. Relative expression analyses of two peroxidase transcripts (PRX52 and PRX2) performed to compare differences among the soybean genotypes indicated that, despite basal levels being similar for the treatments analyzed, tolerant soybeans had a tendency for a higher expression of PRX52 in the presence of aphids. Based on the different patterns observed and the feasibility of analyses performed in this study, enzyme kinetics using V3 infested plants may be a marker for screening RILs in a breeding program targeting the development of aphid-tolerant soybeans.

Natural biological control of Chrysodeixis includens.

A wide variety of abiotic and biotic factors act on insect pests to regulate their populations. Knowledge of the time and magnitude of these factors is fundamental to understanding population dynamics and developing efficient pest management systems. We investigate the natural mortality factors, critical pest stages, and key mortality factors that regulate Chrysodeixis includens populations via ecological life tables. The total mortality caused by natural factors was 99.99%. Natural enemies were the most important mortality factors in all pest stages. The critical stages of C. includens mortality were second and fourth instars. The key mortality factors were predation by ants in the second instar and predation by Vespidae in the fourth instar. The elimination of these factors can cause an increase of 77.52 and 85.17% of C. includens population, respectively. This study elucidates the importance of natural enemies and other natural mortality factors in C. includens population regulation. These factors should be considered in developing and implementing C. includens management strategies and tactics in order to achieve effective and sustainable pest control.

Intraguild Competition of Three Noctuid Maize Pests.

The western bean cutworm Striacosta albicosta (Smith), the fall armyworm Spodoptera frugiperda (J. E. Smith), and the corn earworm Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) are among the major lepidopteran pests of maize in the United States, belonging to the same guild and injuring the reproductive tissues of this crop. Here, intraguild competition of these lepidopterans on non-Bt maize was evaluated through survival analysis of each species under laboratory and field conditions. Competition scenarios were carried out in arenas containing maize silk or ear tissue, using larvae on different stadium of development. Fitness cost competition studies were conducted to examine the influence of intraguild competition and cannibalism and predation rates on larval development. The survival of S. albicosta competing with the other species was significantly lower than in intraspecific competition, even when the larvae were more developed than the competitor. For S. frugiperda, survival remained high in the different competition scenarios, except when competing in a smaller stadium with H. zea Larvae of H. zea had a high rate of cannibalism, higher survival when competing against S. albicosta than S. frugiperda, and reduced survival when the H. zea larvae were at the same development stadium or smaller than the competitors. Based on fitness cost results, the absence of a competitor for the feeding source may confer an advantage to the larval development of S. frugiperda and H. zea Our data suggest that S. frugiperda has a competitive advantage against the other species, while S. albicosta has the disadvantage in the intraguild competition on non-Bt maize.

Plant-to-Plant Movement of Striacosta albicosta (Lepidoptera: Noctuidae) and Spodoptera frugiperda (Lepidoptera: Noctuidae) in Maize ( Zea mays ).

Integrated pest management (IPM) and insect resistance management (IRM) in various cropping systems demand a comprehensive understanding of insect behavior. Among the needed information is basic charaterizations of larval movement and dispersion of some insect-pests, such as the noctuids Striacosta albicosta (Smith) and Spodoptera frugiperda (J.E. Smith). We investigated the plant-to-plant movement of western bean cutworm and fall armyworm larvae in field of maize. Experiments on S. albicosta were conducted between 2008 and 2010. A main study with this pest was performed in 2012 in a randomized complete block design (RCBD) with nine replications. An S. frugiperda study was performed in 2013 in an RCBD with eight replications. The plant-to-plant movement and larval survival were measured in plots with maize nontoxic to the insects. The larval survival of S. albicosta presented high variety throughout the years. Although S. frugiperda survival was relatively low during 2013, it did not compromise the larval assessment. Larvae of both species dispersed governed by nondirectional sensory information, and presented aggregated and symmetrical distribution; however, fall armyworm remained nearer the release point. These results may help the IPM components, such as scouting and economic threshold, as well as the implementation of refuge and seed mixture strategies for IRM.

On-Plant Larval Movement and Feeding Behavior of Fall Armyworm (Lepidoptera: Noctuidae) on Reproductive Corn Stages.

Spodoptera frugiperda J.E. Smith (fall armyworm) is considered one of the most destructive pests of corn throughout the Americas. Although this pest has been extensively studied, little is known about its larval movement and feeding behavior on reproductive compared to vegetative corn stages. Thus, we conducted studies with two corn stages (R1 and R3) and four corn plant zones (tassel, above ear, ear zone, and below ear) in the field at Concord, NE (USA), and in the field and greenhouse at Botucatu, SP (Brazil), to investigate on-plant larval movement. The effects of different corn tissues (opened tassel, closed tassel, silk, kernel, and leaf), two feeding sequence scenarios (closed tassel-leaf-silk-kernel and leaf-silk-kernel), and artificial diet (positive control) on larval survival and development were also evaluated in the laboratory. Ear zone has a strong effect on feeding choice and survival of fall armyworm larvae regardless of reproductive corn stage. Feeding site choice is made by first-instar. Corn leaves of reproductive plants were not suitable for early instar development, but silk and kernel tissues had a positive effect on survival and development of fall armyworm larvae on reproductive stage corn.

Degree-Day Prediction Models for the Flight Phenology of Western Bean Cutworm (Lepidoptera: Noctuidae) Assessed with the Concordance Correlation Coefficient.

Western bean cutworm, Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a native, univoltine pest of corn and dry beans in North America. The current degree-day model for predicting a specified percentage of yearly moth flight involves heat unit accumulation above 10°C after 1 May. However, because the moth's observed range has expanded into the northern and eastern United States, there is concern that suitable temperatures before May could allow for significant S. albicosta development. Daily blacklight moth catch and temperature data from four Nebraska locations were used to construct degree-day models using simple or sine-wave methods, starting dates between 1 January and 1 May, and lower (-5 to 15°C) and upper (20 to 43.3°C) developmental thresholds. Predicted dates of flight from these models were compared with observed flight dates using independent datasets to assess model performance. Model performance was assessed with the concordance correlation coefficient to concurrently evaluate precision and accuracy. The best model for predicting timing of S. albicosta flight used simple degree-day calculations beginning on 1 March, a 3.3°C (38°F) lower threshold, and a 23.9°C (75°F) upper threshold. The revised cumulative flight model indicated field scouting to estimate moth egg density at the time of 25% flight should begin when 1,432 degree-days (2,577 degree-days °F) have accumulated. These results underscore the importance of assessing multiple parameters in phenological models and utilizing appropriate assessment methods, which in this case may allow for improved timing of field scouting for S. albicosta.

Optimization of intrabone delivery of hematopoietic progenitor cells in a swine model using cell radiolabeling with [89]zirconium.

Intrabone (IB) hematopoietic cell transplantation (HCT) of umbilical cord blood in humans remains experimental and the technique has not been optimized. It is unknown whether hematopoietic progenitor cells (HPCs) injected IB are initially retained in the marrow or rapidly enter into the venous circulation before homing to the marrow. To develop an IB-injection technique that maximizes HPC marrow-retention, we tracked radiolabeled human HPCs following IB-injection into swine. We developed a method to radionuclide-label HPCs using a long-lived positron emitter (89) Zr and protamine sulfate that resulted in cellular-retention of low-dose radioactivity. This approach achieved radioactivity levels sufficient for detection by positron emission tomography with both high sensitivity and spatial resolution when fused with computed tomography. We found that conditions utilized in pilot IB-HCT clinical trials conducted by others led to both rapid drainage into the central venous circulation and cellular extravasation into surrounding muscle and soft tissues. By optimizing the needle design, using continuous real-time intra-marrow pressure monitoring, and by reducing the infusion-volume and infusion-rate, we overcame this limitation and achieved high retention of HPCs in the marrow. This method of IB cellular delivery is readily applicable in the clinic and could be utilized in future investigational IB-HCT trials aimed at maximizing marrow retention of HPCs.

Leader-member exchange relationships in health information management.

This article seeks to raise awareness of the leader-member exchange (LMX) theory of leadership and its potential benefit to the health information management (HIM) profession. A literature review that was conducted identified a leadership challenge for HIM practitioners. The review also provides examples of leadership definitions, and potential benefits of LMX to HIM professionals in leading people and influencing leaders in their organizations. The LMX concept may be an avenue to investigate in preparing future and current HIM professionals for leadership.

Characterization of Peroxidase Changes in Tolerant and Susceptible Soybeans Challenged by Soybean Aphid (Hemiptera: Aphididae).

Changes in protein content, peroxidase activity, and isozyme profiles in response to soybean aphid feeding were documented at V1 (fully developed leaves at unifoliate node, first trifoliate leaf unrolled) and V3 (fully developed leaf at second trifoliate node, third trifoliate leaf unrolled) stages of soybean aphid-tolerant (KS4202) and -susceptible (SD76R) soybeans. Protein content was similar between infested and control V1 and V3 stage plants for both KS4202 and SD76R at 6, 16, and 22 d after aphid introduction. Enzyme kinetics studies documented that control and aphid-infested KS4202 V1 stage and SD76R V1 and V3 stages had similar levels of peroxidase activity at the three time points evaluated. In contrast, KS4202 aphid-infested plants at the V3 stage had significantly higher peroxidase activity levels than control plants at 6 and 22 d after aphid introduction. The differences in peroxidase activity observed between infested and control V3 stage KS4202 plants at these two time points suggest that peroxidases may be playing multiple roles in the tolerant plant. Native gels stained for peroxidase were able to detect differences in the isozyme profiles of aphid-infested and control plants for both KS4202 and SD76R.

Antibiotic resistance gene profiling of faecal and oral anaerobes collected during an antibiotic challenge trial.

Here we describe a study examining the antibiotic resistance gene carriage in anaerobes collected during a clinical study. The results demonstrated that genes normally associated with anaerobes were most prevalent such as tetQ, cepA and cblA although several genes associated with Enterobacteriaceae including sul2, blaSHV and strB were also detected.

Western bean cutworm survival and the development of economic injury levels and economic thresholds in field corn.

Western bean cutworm, Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a native pest of dry beans (Phaseolus vulgaris L.) and corn (Zea mays L.). Historically, the western bean cutworm was distributed in the western United States, but since 1999 eastward expansion has been observed. In corn, economic impact is caused by larval ear feeding. Information on western bean cutworm biology, ecology, and economic impact is relatively limited, and the development of economic injury levels (EILs) and economic thresholds (ETs) is required for more effective management. Studies during 2008-2011, across three ecoregions of Nebraska, sought to characterize western bean cutworm survival and development of EILs and ETs. Calculations of EILs and ETs incorporated the dynamics of corn price, management cost, and pest survival. The results from the current study demonstrated low larval survival of this species (1.51-12.82%). The mean yield loss from one western bean cutworm larva per plant was 945.52 kg/ha (15.08 bu/acre), based on 74,100 plants per ha. Economic thresholds are expressed as a percentage of plants with at least one egg mass. This study is the first study that explicitly incorporates variable management costs and crop values into western bean cutworm EIL calculations, and larval survival into ET calculations.

Evaluation of late vegetative and reproductive stage soybeans for resistance to soybean aphid (Hemiptera: Aphididae).

The soybean aphid, Aphis glycines Matsumura, has become the most significant soybean [Glycine max (L.) Merrill] insect pest in the north central soybean production region of North America. The objectives of this research were to measure selected genotypes for resistance to the soybean aphid in the later vegetative and reproductive stages under field conditions, and confirm the presence of tolerance in KS4202. The results from 2007 to 2011 indicate that KS4202 can support aphid populations with minimal yield loss at levels where significant yield loss would be expected in most other genotypes. The common Nebraska cultivar, 'Asgrow 2703', appears to show signs of tolerance as well. None of the yield parameters were significantly different between the aphid infested and noninfested treatments. Based on our results, genotypes may compensate for aphid feeding in different ways. Asgrow 2703 appears to produce a similar number of seeds as its noninfested counterpart, although the seeds produced are slightly smaller. Field evaluation of tolerance in KS4202 indicated a yield loss of only 13% at 34,585-53,508 cumulative aphid-days, when 24-36% yield loss would have been expected.

Comparative evaluation of the potency and antigenicity of two distinct BoNT/A-derived formulations.

IncobotulinumtoxinA (Xeomin®) and onabotulinumtoxinA (BOTOX®) are unique botulinum neurotoxin type A (BoNT/A)-derived drugs. IncobotulinumtoxinA utilizes the naked 150 kDa holotoxin portion of BoNT/A, whereas onabotulinumtoxinA uses the complete native 900 kDa complex as drug substance. On the basis of purportedly similar pharmacological characteristics, these formulations were evaluated for potency by LD50 and mouse Digit Abduction Score (DAS) bioassays. DAS was also used to assess antigenicity. Full-range DAS dose-response profiles were achieved with four lots of each product, with similar observations between lots for a given product. Between products, however, the mean DAS potency of incobotulinumtoxinA (ED50 range 7.0-10.2 U/kg) was significantly lower than that of onabotulinumtoxinA (ED50 range 4.4-6.4 U/kg), consistent with lower measured potencies in the LD50 assay for incobotulinumtoxinA (potency range 62-82 U). In assessments of DAS duration of effect at similar unit doses, the observed lower potency of incobotulinumtoxinA translated into decreased peak efficacy and dose effect over time (i.e. shorter duration). In contrast, at equi-efficacious doses yielding near-maximal DAS responses, both toxin formulations were uniformly inhibited in a statistically significant manner when preincubated with rabbit-derived, onabotulinumtoxinA-neutralizing antibodies, supporting the position that inhibition of 150 kDa holotoxin serves as the common basis for neutralization and, therefore, incobotulinumtoxinA would not be expected to be effective in onabotulinumtoxinA-immunoresistant subjects (and vice versa). Further, with lower lot-to-lot relative potency, incobotulinumtoxinA is not dose-equivalent or interchangeable with onabotulinumtoxinA, suggesting that various aspects of drug product formulation may influence observed pharmacology.