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Our objectives were to examine changes in endometrial and luteal gene expression during estrus, diestrus, pregnancy and treatments to induce luteolysis and putatively induce luteostasis. Groups were: Diestrus (DIEST), Estrus (ESTR), Pregnant (PREG), Oxytocin (OXY), Carbetocin (CARB), and Meclofenamic acid (MFA). Blood was obtained from day (D)12 to D15 for measurement of oxytocinase, also referred to as leucyl-cysteinyl aminopeptidase (LNPEP) and progesterone. Luteal biopsies were obtained on D12 and D15 and an endometrial biopsy on D15. Real-time RT-PCR was performed for the following genes: PGR, ESR1, OXTR,OXT, LNPEP, PTGS2, PTGFR, PLA2G2C, PTGES, SLC2A4, and SLC2A1. Regarding serum LNPEP, PREG and OXY (p-value0.001) had higher concentrations than DIEST mares. Endometrial PTGES expression was higher (p-value0.04) in DIEST, PREG and OXY than other groups. Endometrium from ESTR had increased expression of OXT (p-value0.02) compared to MFA and OXY mares. Carbetocin treatment: decreased serum progesterone and LNPEP; increased endometrial PLA2G2C; decreased endometrial PTGES; and decreased luteal aromatase and PTGES. Treatment with MFA: decreased endometrial PLA2G2C, increased endometrial PTGES; and resulted in less OXTR and OXT luteal abundance on D12 compared to D15. Endometrial and luteal expression of LNPEP is affected by physiologic stage and treatment and is involved in luteal function and pregnancy recognition pathways through effects on oxytocin and prostaglandin synthesis in the horse.

The development and application of new molecular tools has shown the real complexity involved in Mycobacterium Tuberculosis infection. The sequencing of the complete Koch's Bacillus genome can reveal information about the true extent of the point mutations that confer resistance to treatment to these bacteria. Complete genome sequencing techniques and their availability in public databases, have been responsible for the exponential growth of computational tools to extract information about important features encoded in the genomes of Mycobacterium complex species. This is a literature review study, from scientific research platforms, with the aim of discussing how computational tools used for analysis of the complete genome sequencing of M. tuberculosis can identify the genetic relationship between strain mutations and antimicrobial resistance. Whole genome sequencing is considered a surveillance marker of transmission and drug resistance, as well as a strong predictor of the emergence of possible resistance mechanisms for first- and second-line drugs used in the treatment of pulmonary tuberculosis.

Introduction: The recombinant factor FVIII Fc fusion protein (rFVIIIFc) is a first-in-class extended half-life FVIII product to treat patients with hemophilia A. The safety, efficacy and prolonged half-life of rFVIIIFc was demonstrated in the phase 3 studies A-LONG, Kids A-LONG and the extension study ASPIRE. Despite robust efficacy and safety data of rFVIIIFc therapy from clinical trials, evidence on the effectiveness of rFVIIIFc use in real-world remains scarce. Our analysis aimed at investigating the effectiveness of prophylactic rFVIIIFc treatment in routine clinical use in Germany. Material and Methods: Twenty-seven patients with severe hemophilia A, who switched from prophylaxis with conventional recombinant factor VIII (rFVIII) products to rFVIIIFc, were included. Annualized bleeding rates, factor consumption, number of injections and adherence to prophylaxis were compared. The retrospective period prior switching to rFVIIIFc was three years, while the mean follow-up period after switching to rFVIIIFc was 24.9 months. Results: Switching to rFVIIIFc led to a 33.7% reduction in mean annualized number of injections and a 18.3% reduction in mean annualized factor consumption while maintaining low bleeding rates. The mean annualized bleeding rate (ABR) was 2.5 and 1.7 for rFVIII and rFVIIIFc, respectively. The adherence improved from 87% to 94%. During the follow-up period eleven surgeries were performed; all with a hemostatic response rated as excellent. No FVIII inhibitor formation after switching to rFVIIIFc has been detected. Conclusion: Real-world treatment with rFVIIIFc was associated with substantial reductions in consumption and injection frequenies while maintaining low bleeding rates supporting safety and efficacy data from clinical trials.

Heat stress is a major concern in animal reproduction, as testicular temperature must be 3–5 °C below body core temperature for production of motile and fertile sperm in mammals. Although recent studies concluded that increased temperature per se was the underlying pathophysiology of testicular impairment, more studies are required to better understand the mechanisms. Therefore, our objective was to investigate the impacts of mild acute heat stress on sperm and testes, and based on mRNA, elucidate involvement of StAR, Trp53 and Trp53-dependent intrinsic and extrinsic apoptotic pathways in pathophysiology of testicular heat stress. Forty-eight C57 BCL6 elite male mice were equally allocated into six groups, anesthetized and the distal third of their body immersed in a water-bath at 40 or 30 °C (heat treatment and control, respectively) for 20 min. Intervals from heat exposure (Day 0) to euthanasia were: 8 and 24 h and 7, 14 and 21 d (plus a control group at 14 d). The epididymides were excised, minced and placed in Tyrode albumin lactate pyruvate hepes (TALPH) at 37 °C for 15 min to recover sperm. Based on computer assisted sperm analysis (CASA), heat treatment reduced total and progressive motility ∼40% (P < 0.05) on Days 14 and 21. Furthermore, percentage morphologically normal sperm was significantly decreased on Day 7, with greater reductions on Days 14 and 21, mostly due to increased midpiece defects. Acrosome integrity (FITC PSA) was decreased ∼35% at 8 h (P < 0.05) and reached a nadir on Day 14. There were decreases (P < 0.05) in seminiferous tubule diameter and testicular weight (relative to body weight) on Day 14. Testicular RNA was extracted, reverse-transcribed and cDNA used for PCR. Expression of genes Hspa1b (Hsp70) and Gpx1 had 7- and 10-fold increases (P < 0.001 for each) at 8 and 24 h, respectively, with Hspa1b remaining upregulated at 24 h, whereas StAR peaked at Day 14 (15-fold, P < 0.0001) and had returned to baseline on Day 21. Both Trp53 and Casp8 were upregulated (P < 0.05) on Day 14, whereas Bcl-2 was decreased (P < 0.05) on Days 7 and 14. In conclusion, acute mild heat stress severely reduced sperm quality and based on mRNA, there was upregulation of chaperone and antioxidant systems and Trp53-dependent intrinsic and extrinsic apoptotic pathways, with deleterious effects on sperm, spermatocytes and spermatids. These findings provided insights into the pathophysiology of heat stress and should contribute to development of evidence-based approaches to mitigate effects of testicular heating.

Claudia Klein ist Dozentin an einer Berufsfachschule für Ergotherapie und dort unter anderem verantwortlich für den Bereich „Digitale Medien“. In diesem Rahmen stellte sie sich die Frage, welche Kompetenzen die Lehrenden im Umgang mit digitalen Medien besitzen und welchen Stellenwert die Digitalisierung in Ausbildung und Studium einnimmt.

In patients with haemophilia (PwH), most frequently affected joints are the ankle, knee and elbow. Due to improved factor therapy in the last decades, these previous findings have to be verified in Germany.The aim of this study is to detect the most affected joint, evaluate the significance of the source of pain and determine the point prevalence of back pain in Germany today.In a retrospective study, data of n = 300 patients with severe moderate and mild haemophilia were evaluated regarding the most affected joint, the most common source of pain, and the point prevalence of back pain. An anamnesis questionnaire and the German Pain Questionnaire were used for this assessment.The most affected joint in German PwH is still the ankle (41%), followed by the knee (27%) and the elbow (11%). The most common source of pain is also the ankle joint (32%). Back pain was also identified as one of the most common sources of pain, which is comparable to the elbow (elbow:15%; back:13%). The point prevalence in PwH for back pain was significantly higher compared to the general German population (P = .031).Our data showed that the ankle is still the most affected joint and the most common source of pain in Germany. These results also showed the relevance of back pain as a pain source. The evaluations also demonstrated the high point prevalence of back pain in PwH. Future therapies should also focus on the spine because joint changes affect posture.

We report a case of bilateral and acute angle closure after a single dose of antidepressant venlafaxine in a 40-year-old woman with no previous pathologies, who asked for consultation for blurred vision and pain in the left eye. Initial evaluation included visual acuity, slit lamp biomicroscopy and intraocular pressure (IOP) measurement using Goldmann’s applanation tonometer. Gonioscopy and fundus examination were also performed in both eyes. Examination and IOP supported the diagnosis of acute glaucoma in the left eye. The patient’s evolution was satisfactory after bilateral peripheral iridotomy was performed with Nd-YAG laser, as described in the cases reported in the international literature. The pathophysiology of angle closure and its relationship with venlafaxine intake were also discussed.

Veränderungen finden ständig statt – egal, ob in der Politik, Mode oder Technik. Seit ein bis zwei Jahren macht besonders eine Entwicklung Schlagzeilen: die viel diskutierte, manchmal belächelte und oft kritisierte Generation Z. Mit ihren Werten und Einstellungen stellt sie Arbeitgeber, Kollegen und Lehrende vor neue Herausforderungen.

Aluteal cycles were induced in the mare to evaluate the effects of progesterone deprivation on the gene expression of embryos and endometrium collected eight days after ovulation. We hypothesized that the transcript expression would be altered during induced aluteal (AL) cycles (low progesterone 4 ng/mL) for 1) the embryonic expression of progesterone-mediated transcripts and those related to normal embryo growth and development and 2) the endometrial expression of progesterone-mediated transcripts and those related to prostaglandin synthesis and normal pregnancy establishment. Seven cyclic mares with a median age of 6.5 years (range 3–16) were utilized in a crossover design. Mares in estrus were artificially inseminated to a fertile stallion and randomly assigned to control or AL groups. Mares received either saline solution (control mares) or PGF2α (AL mares), twice daily on days 0, 1, and 2 and once daily on days 3 and 4. Serial blood samples were collected daily from day 0 (ovulation) until the day of embryo collection and endometrial biopsy on day 8. Mares were monitored until they returned to estrus, and artificially inseminated. Mares were switched to the opposite treatment group only after a successful embryo collection occurred during the previous cycle and only cycles that produced embryos were used for analyses. The study design resulted in paired samples from each mare for analyses. No significant rise in progesterone was observed in the AL group with mean concentrations of plasma progesterone remaining

Our understanding of the temporal changes in endometrial and luteal gene transcripts related to the actions of oxytocin and prostaglandin during early equine pregnancy is incomplete. Additionally, the role of oxytocinase, also known as Leucyl-cystinyl aminopeptidase (LNPEP), during early pregnancy in mares has not been previously investigated. Luteal and endometrial biopsies were obtained on Day (D)8, D10, D12 and D15 post-ovulation in pregnant (PREG) and diestrus (DIEST) mares for real-time qPCR. Differences in endometrial gene expression occurred over time in: SLC2A4, SLC2A1, PTGES, OXTR and LNPEP. PTGFR and PLA2G2C had lower relative abundance in PREG D15 endometrium compared to D10. OXT and OXTR were increased on D10 and 15 PREG, respectively. Regarding luteal mRNA relative abundance, ESR1, PTGS2, PTGFR, and PTGES had higher relative abundance in D12 of DIEST and PREG. Luteal expression of OXTR and OXT had higher relative abundance in D15 compared to D8, and LNPEP had higher relative abundance in D10 and 12. Endometrial and luteal PTGES had an increased mRNA abundance in both D12 DIEST and PREG mares, which may lead to additional luteoprotective prostaglandin E2 (PGE2) secretion. Furthermore, luteal SLC2A1 had higher relative abundance in pregnancy, and likely supports the high metabolic activity of luteal tissue by increasing glucose uptake. Oxytocinase is present in endometrial and luteal tissue and its role in oxytocin induced prostaglandin secretion is uncertain.

Maternal recognition of pregnancy (MRP) resulting in prolonged luteal lifespan is crucial to maintenance of pregnancy. Mechanisms underlying MRP differ in between species, whereat the precise mechanism by which luteolysis is prevented has yet to be described for equine pregnancy. Transcriptional profiling studies entailing analysis of endometrial and embryonic tissues have shed light into the wider context of maternal recognition of pregnancy. Our own studies revealed the secretion of fibrinogen by preimplantation equine embryos, which is a peculiar finding as fibrinogen synthesis almost exclusively occurs in the liver. Conceptus-derived fibrinogen likely contributes to cell adhesion during fixation and underlines the helpfulness of transcriptome studies in identifying species-specific aspects of early pregnancy. Despite all our advances, we yet have to decipher the signaling pathways leading to reduced endometrial production of prostaglandin F2alpha during MRP. We recently analyzed samples from mares undergoing spontaneous pregnancy loss in the face of high progesterone in comparison to samples collected from mares at Day 20 of intact pregnancy. No differentially expressed transcripts could be detected. It appears that after maternal recognition of pregnancy and consequent suppression of luteolysis, progesterone, not the presence of a viable conceptus, is the driving force of gene expression. This finding underlines the importance of understanding the events surrounding luteostasis. We propose that a standardized approach in vitro cell culture model is needed to address mechanistic questions pertaining to MRP. Approaches currently underway such as CRISPR/Cas9-mediated targeted integration of TERT and TERC to immortalize equine endometrial stromal fibroblasts will be discussed.

Human male reproductive development has a prolonged prepubertal period characterized by juvenile quiescence of germ cells with immature spermatogonial stem cell (SSC) precursors (gonocytes) present in the testis for an extended period of time. The metabolism of gonocytes is not defined. We demonstrate with mitochondrial ultrastructure studies via TEM and IHC and metabolic flux studies with UHPLC-MS that a distinct metabolic transition occurs during the maturation to SSCs. The mitochondrial ultrastructure of prepubertal human spermatogonia is shared with prepubertal pig spermatogonia. The metabolism of early prepubertal porcine spermatogonia (gonocytes) is characterized by the reliance on OXPHOS fuelled by oxidative decarboxylation of pyruvate. Interestingly, at the same time, a high amount of the consumed pyruvate is also reduced and excreted as lactate. With maturation, prepubertal spermatogonia show a metabolic shift with decreased OXHPOS and upregulation of the anaerobic metabolism-associated uncoupling protein 2 (UCP2). This shift is accompanied with stem cell specific promyelocytic leukemia zinc finger protein (PLZF) protein expression and glial cell-derived neurotropic factor (GDNF) pathway activation. Our results demonstrate that gonocytes differently from mature spermatogonia exhibit unique metabolic demands that must be attained to enable their maintenance and growth in vitro.

Background Record keeping is integral to home treatment for haemophilia. Issues with paper diaries include questionable compliance, data validity and quality. Implementation of electronic diaries (e-diaries) in haemophilia patients could improve documentation of home treatment. Aim This article evaluates the effects of an e-diary, Haemoassist, on recording and patient compliance with therapy. Patients and Methods An explorative study was used to assess the sequential use of paper diaries and e-diaries by 99 patients with severe haemophilia A or B and 1 with severe factor VII deficiency. Median age was 41 years. Information was obtained from paper records for 3 years preceding the introduction of an electronic record system and the first 6 to 12 months of Haemoassist use. Data from the 3-year period were averaged. Missing data for rounded 12 months of e-diary use were extrapolated to correspond to a full year. Results Enhancement of 23% in record delivery was observed for the period of Haemoassist use (p = 0.013). Twenty-one percent increase in patients’ compliance for data reporting (from 65% 35 to 86% 22, p = 0.003) and 16% increase for documentation of bleedings (from 68 to 84% of patients, p = 0.01) were detected. Compliance to prescribed therapy of patients for the whole studied period improved by 6% (from 82% ± 29 to 88% ± 25, p = 0.05). Major advances were demonstrated predominantly in the age groups of between 13 and 20 and 21 and 40 years. Conclusion e-Diaries' use enables improved recording of information about patients' home treatment and bleeding episodes. Enhanced compliance with therapy may be a further benefit.

A produção de milho se destaca no Brasil como o principal cereal cultivado e se torna cada vez mais importante devido ao aumento da demanda de produção de alimentos. A adubação nitrogenada, além de possuir alto custo no cultivo do milho, tem grande influência nos componentes de rendimento da cultura. Pela dinâmica do nitrogênio ser muito dependente das condições climáticas se buscam fontes alternativas para diminuir perdas no sistema. Sendo assim, o objetivo desta pesquisa foi avaliar a eficiência agronômica da cultura do milho, sendo este submetido a diferentes fontes de nitrogênio em cobertura, buscando testar as principais fontes do nutriente disponíveis no mercado. O delineamento experimental utilizado foi de blocos casualizados, com cinco blocos e cinco tratamentos. Foi utilizada a espécie híbrido comercial 30f53 VYHR e os tratamentos consistiram de testemunha, Nitromag, Super N, Cooper N e Ureia. Foram analisadas as variáveis: índice de clorofila foliar, rendimento de grãos e produção de massa verde e seca. Os resultados foram submetidos à análise de variância pelo teste F e as diferenças entre médias foram comparadas pelo teste de Tukey (P≤0,05) de probabilidade de erro. As diferentes fontes de N não diferiram, significativamente, entre si para o índice de clorofila foliar, rendimento de grãos e produtividade de massa verde e seca.

O presente estudo teve por objetivo avaliar arranjos espaciais na cultura da soja, no município de Anchieta (SC). O delineamento experimental utilizado foi de blocos casualizados com cinco repetições e quatro tratamentos (0,60 m, 0,45 m, 0,30 m e 0,20 x 0,40 m-fileiras duplas), utilizando a cultivar AMS Tibagi RR. Avaliou-se a altura de inserção da primeira vagem, massa de mil grãos, produtividade real e potencial. Os dados foram submetidos à análise de variância pelo teste F e quando significativos, comparados pelo teste de Tukey a 5% de probabilidade de erro. A cultura da soja submetida a diferentes arranjos espaciais não apresentou diferença estatística para as variáveis produtividade real, potencial e massa de mil grãos. A diferença estatística foi constatada para a variável altura de inserção da primeira vagem, sendo que, quanto mais próximos os espaçamentos, maior a altura de inserção da primeira vagem. Portanto, diferentes espaçamentos não influenciam a produtividade de grãos de soja.

Interferon epsilon (IFNE) is type I interferon which stands out through its unusual expression profile and differing regulation compared to classic type I interferons such as interferon alpha and interferon beta. Unlike other type I interferons, the expression of IFNE is not stimulated through exposure to viral agents. Expression of IFNE is most abundant in mouse and human endometrium where it is constitutively expressed in luminal and glandular epithelial cells and expression levels are up-regulated with estrogen exposure. The aim of the current study was to determine whether a cycle or pregnancy dependent expression pattern of IFNE is existent in equine endometrium and to localize IFNE expression within the endometrium. Additionally, endometrial explant culture and culture of mixed epithelial/stromal cells populations was used to determine the effects estrogen and seminal plasma on IFNE transcript abundance. Samples collected during diestrus and pregnancy expressed significantly higher levels of IFNE than samples obtained from anestrous or estrous mares (P 0.001). Exposure of mixed endometrial epithelial/stromal cell populations and endometrial explants to 10% seminal plasma and estradiol 17-beta did not affect IFNE expression levels (P 0.05). Upon in situ hybridization, staining was exclusively present in luminal and glandular epithelial cells, with stromal displaying absent staining intensity. Both diestrous and pregnant samples were characterized by markedly stronger staining of glandular epithelial cells than anestrous and estrous samples. The progesterone-dependent increase in IFNE abundance during the estrous cycle likely implies that IFNE is part of the innate immune system in endometrium that gives protection against uterine infections during progesterone-dominated phase of the estrous cycle.

International audience; A key element to understanding parasite epidemiology is assessing their prevalence in the respective wild reservoir hosts. The tapeworm Echinococcus multilocularis circulates between canid species (definite hosts) and small mammals (mostly rodents; intermediate hosts). Prevalence rates of Echinococcus multilocularis in the intermediate host are most exclusively determined through macroscopic examination of the liver generally followed by molecular or histological diagnostic for parasite species confirmation. The overall objective of the study was to investigate the suitability of Real-Time PCR and Droplet Digital PCR (ddPCR) analysis as tool to detect exposure pressure (frequency of infection events) from E. multilocularis in intermediate hosts even in the absence of macroscopic lesions in the liver. One hundred six small mammals (meadow voles and deer mice) were trapped followed by post-mortem examination including macroscopic evaluation of the liver to detect lesions indicative of infection with Echinococcus multilocularis but also by sampling a piece of liver in absence of lesion to submit it to molecular assay. Macroscopic lesions were present in the livers of two samples. Including the latter two samples, five samples yielded a positive result following Real-Time PCR, whereas 16 samples displayed three or more positive droplets upon ddPCR and were considered positive. Whether these additional cases without macroscopic lesions would have become infectious during the lifespan of the rodent or were abortive or early infections is unclear, but these data suggest levels of exposure of intermediate hosts to the parasite is much higher than assumed.

The current study evaluated equine endometrial explants following 12, 24, and 48 hours in culture. Measurement of an indicator of cell death in explant supernatant, light microscopy, and gene expression of biomarkers of endometrial function and cellular stress, were used to compare the effect of six different media on explant viability and morphology. Viability of explants was assessed indirectly through measuring LDH activity in the culture supernatant. Regardless of culture medium composition, a significant increase in LDH activity was observed within 12 hours of culture, indicating occurrence of cell damage. Morphological analysis through light microscopy revealed degenerative changes occurring within 12 hours and after 48 hours there is nearly complete loss of luminal and superficial glandular epithelium and diffuse detachment of deep glandular epithelium. Transcript abundance of prostaglandin-endoperoxide synthase 2 (PTGS2), estrogen receptor 1 (ESR1), and vascular endothelial growth factor (VEGF) was assessed as biomarkers of endometrial function. A marked increase in PTGS2 and VEGF expression occurred, ESR1 displayed more or less steady expression levels. Above described changes were seen irrespective of cell culture medium used. The marked increase in expression in PTGS2 expression presents a limitation to using endometrial explants in the current culture system to study aspects of endometrial function such as the inflammatory response to insemination. This article is protected by copyright. All rights reserved.