Miracle-dz/UniFAR-FTUs huggingface dataset:
Normal Fertility Requires the Expression of Carbonic Anhydrases II and IV in Sperm*

HCO3− is a key factor in the regulation of sperm motility. High concentrations of HCO3− in the female genital tract induce an increase in sperm beat frequency, which speeds progress of the sperm through the female reproductive tract. Carbonic anhydrases (CA), which catalyze the reversible hydration of CO2 to HCO3−, represent potential candidates in the regulation of the HCO3− homeostasis in sperm and the composition of the male and female genital tract fluids. We show that two CA isoforms, CAII and CAIV, are distributed along the epididymal epithelium and appear with the onset of puberty. Expression analyses reveal an up-regulation of CAII and CAIV in the different epididymal sections of the knockout lines. In sperm, we find that CAII is located in the principal piece, whereas CAIV is present in the plasma membrane of the entire sperm tail. CAII and CAIV single knockout animals display an imbalanced HCO3− homeostasis, resulting in substantially reduced sperm motility, swimming speed, and HCO3−-enhanced beat frequency. The CA activity remaining in the sperm of CAII- and CAIV-null mutants is 35% and 68% of that found in WT mice. Sperm of the double knockout mutant mice show responses to stimulus by HCO3− or CO2 that were delayed in onset and reduced in magnitude. In comparison with sperm from CAII and CAIV double knockout animals, pharmacological loss of CAIV in sperm from CAII knockout animals, show an even lower response to HCO3−. These results suggest that CAII and CAIV are required for optimal fertilization.
WDR82, a Key Epigenetics-Related Factor, Plays a Crucial Role in Normal Early Embryonic Development in Mice1

Epigenetic regulation is considered one of the most important mechanisms by which changes in gene expression occur without changes in the underlying DNA sequence. More and more studies have shown that this kind of regulation plays a very important role during the process of early embryonic development. Methylation of histones is a special process in epigenetic regulations that plays a dual role: some activate gene expression, while some inhibit it; trimethylation of histone 3 lysine 4 has been shown to be a marker of gene expression activation. Previous research has led us to focus on the role of WDR82, which has been shown to recognize a subunit in the methyltransferases complex that catalyzes H3K4me3 in early embryonic development. Although it has been shown that a defect in WDR82 causes dysfunction of SETD1A/SETD1B and results in loss of H3K4me3 in human cell lines, the exact role of WDR82 in the methyltransferases complex during early embryonic development is not clear. Our study has shown that a defect in WDR82 causes dysfunction of SETD1A/SETD1B and affects the normal H3K4me3 status in the transcription start region of POU5F1, which then causes the down-regulation of POU5F1 as well as its downstream factors STAT3/BIRC5, which are responsible for the extremely high apoptotic rates of blastocysts. Finally, the result of a blocked WDR82 consists of stunted embryonic development and death. Thus, WDR82 can be considered a key epigenetic regulation-related factor crucial in the normal growth and development of embryos.
Equine CRISP3 Modulates Interaction Between Spermatozoa and Polymorphonuclear Neutrophils1

Equine spermatozoa induce a uterine inflammatory response characterized by a rapid, transient influx of polymorphonuclear neutrophils (PMNs). Seminal plasma proteins have been shown to modulate the interaction between spermatozoa and PMNs, but a specific protein responsible for this function has not been identified. The objective of this study was to isolate and identify a protein in equine seminal plasma that suppresses binding between spermatozoa and PMNs. Seminal plasma was pooled from five stallions, and proteins were precipitated in 60% (w/v) ammonium sulfate and dialyzed (3500 MW cutoff). Proteins were submitted to a Sephacryl S200 column, and fractions were pooled based on the fraction pattern. Each pool was analyzed for protein concentration and tested for its suppressive effect on PMN/sperm binding. Protein pools with biological activity were submitted to ion-exchange chromatography (diethylaminoethyl [DEAE] Sephadex column) with equilibration buffers containing 0.1–0.5M NaCl. Eluants were pooled, analyzed for protein concentration, and tested for suppressive effects on PMN/sperm binding. Protein distribution and purity were determined by one- and two-dimensional SDS-PAGE, and the purified protein was submitted for sequence analysis and identification. This protein was identified as equine CRISP3 and was confirmed by Western blotting. Suppression of PMN/sperm binding by CRISP3 and seminal plasma was confirmed by flow cytometry (22.08% ± 3.05% vs. 2.06% ± 2.02% vs. 63.09% ± 8.67 for equine seminal plasma, CRISP3, and media, respectively; P < 0.0001). It was concluded that CRISP3 in seminal plasma suppresses PMNs/sperm binding, suggesting that CRISP3 regulates sperm elimination from the female reproductive tract.
Competence of the housefly, Musca domestica, as a vector of Microsporum canis under experimental conditions

Abstract The role of Musca domestica Linnaeus as a vector of the dermatophyte Microsporum canis was investigated under experimental laboratory conditions. About 400 4‐day‐old M. domestica flies were divided into two groups. Group A consisted of about 200 infected flies and group B comprised about 200 uninfected flies that were used as controls. Each trial was run three times. Flies from group A were fed for 24 h with a solution of ultra‐high temperature‐treated (UHT) milk containing about 106 colony‐forming units (CFU) per mL of M. canis (infected milk inoculum [IMI]). The control group (group B) was fed with only UHT milk spiked with a teaspoon of honey. Microsporum canis was detected from faeces, vomitus, external surfaces and internal organs of 20 adult flies, eggs, first‐, second‐ and third‐stage (L1, L2, L3) larvae and pupae of each group, as well from 20 adult newly emerged flies (NEFs; from infected generations only). Samples were collected at 2, 4, 6 and 24 h post‐infection (p.i.) (i.e. the times at which IMI was available) and on 2, 5, 7 and 8 days p.i. from adult flies, faeces and vomitus. Eggs, L1, L2, L3 and pupae were processed as soon as they appeared. Equivalent samples were taken from group B. All the samples were individually cultured. Microsporum canis was not isolated from the control group, from eggs, larvae, pupae or NEFs, or from faeces and vomitus, although it was detected on the body surface (26.2%) and internal organs (26.9%) of adult flies. The highest positivity for M. canis was detected on flies within the first 6 h p.i. (i.e. 57.2% on the body surface and 71.6% in the internal organs). No M. canis was isolated at 24 h p.i. , but it was isolated from the body surface only at 2 and 5 days p.i. The results presented provide evidence that M. domestica transmits M. canis mechanically with its outer body surface for up to 5 days p.i. , but does not do so through its vomitus and faeces or transovarially. The role played by M. domestica in the epidemiology of human and animal dermatophytoses is discussed.
Inhibition of Class IIb Histone Deacetylase Significantly Improves Cloning Efficiency in Mice1

Since the first mouse clone was produced by somatic cell nuclear transfer, the success rate of cloning in mice has been extremely low. Some histone deacetylase inhibitors, such as trichostatin A and scriptaid, have improved the full-term development of mouse clones significantly, but the mechanisms allowing for this are unclear. Here, we found that two other specific inhibitors, suberoylanilide hydroxamic acid and oxamflatin, could also reduce the rate of apoptosis in blastocysts, improve the full-term development of cloned mice, and increase establishment of nuclear transfer-generated embryonic stem cell lines significantly without leading to obvious abnormalities. However, another inhibitor, valproic acid, could not improve cloning efficiency. Suberoylanilide hydroxamic acid, oxamflatin, trichostatin A, and scriptaid are inhibitors for classes I and IIa/b histone deacetylase, whereas valproic acid is an inhibitor for classes I and IIa, suggesting that inhibiting class IIb histone deacetylase is an important step for reprogramming mouse cloning efficiency.
Effects of Genetic Captive-Breeding Protocols on Sperm Quality and Fertility in the White-Footed Mouse1

Mice (Peromyscus leucopus noveboracensis) from a captive-breeding program were used to test the effects of three genetic breeding protocols (minimizing mean kinship [MK], random breeding, and selection for docility [DOC]) and inbreeding levels on sperm traits and fertility. Earlier, in generation 8, one DOC replicate went extinct because of poor reproductive success. By generation 10, spermatozoa from DOC mice had more acrosome and midpiece abnormalities, which were shown to be strong determinants of fertility, as well as lower sperm production and resistance to osmotic stress. In addition, determinants of fertility, including male and female components, were assessed in a comprehensive manner. Results showed that the probability (P) of siring litters is determined by sperm number, sperm viability, and midpiece and acrosome abnormalities; that the P of siring one versus two litters is determined by tail abnormalities; and that the total number of offspring is influenced by female size and proportion of normal sperm, showing the relative importance of different sperm traits on fertility. On average, males with 20% normal sperm sired one pup per litter, and males with 70% normal sperm sired eight pups per litter. Interestingly, the proportion of normal sperm was affected by docility but not by relatively low inbreeding. However, inbreeding depression in sperm motility was detected. In the MK group, inbreeding depression not only affected sperm motility but also fertility: An increase in the coefficient of inbreeding (f) of 0.03 reduced sperm motility by 30% and translated into an offspring reduction of three pups in second litters. A genetic load of 48 fecundity equivalents was calculated.
Epidemiological Analysis and Cost of Hospitalization Associated with Pediatric Burns in Kermanshah, Iran

Background: burn injuries are a major public health in the world, especially in developing countries. The aim of this study was investigate to the epidemiological data and cost of hospitalization due to burn in pediatrics in Kermanshah provinces from 2011 to 2013.Material and Methods: this was a retrospective cross sectional study. The study subject was all of patient lee than 15 years who admitted in Burns Center at Imam Khomeini Hospital in Kermanshah, Iran, from 21 March 2011 to March 2013 (two years). The data including age, gender, cause of burn, burn degree, place of burn, length of stay (LOS), burned body surface (BBS) and cost of hospitalization was obtained from hospital data and analyzed by SPSS version 18. Results: the overall mean age was 5.27 ± 4.52 years; the ages ranged from less than 1 year to 15 years. The mean BBS % and LOS was 22.8 % and 7.48 day, respectively. The mean cost per patient, per hospitalization day and per % BBS were 15000000 IRR, 657981 IRR and 20045348 IRR, respectively. Conclusion: The current study showed the main cause of burn and mortality in the pediatric population was hot liquids and flame, respectively. An important point is that most of burn injuries in pediatric population are preventable and avoidable if the necessary training about cause of these burns provides for their parent.
Biomarkers for Diagnosis of Axial Spondyloarthritis, Disease Activity, Prognosis, and Prediction of Response to Therapy

There exists a major unmet need for biomarkers that can identify axial spondyloarthritis (axSpA) early after disease onset because of the availability of highly effective therapies. Several recent reports have examined the autoantibody response in patients with axSpA through the use of protein microarrays and protein-protein interactions although diagnostic performance of biomarkers identified to date has been inadequate. An example of such a biomarker is protein phosphatase magnesium-dependent 1A. Antibodies to the human leukocyte antigen class II-associated invariant chain peptide (anti-CD74) are candidate diagnostic biomarkers but sensitivity declines with increasing duration of disease. Metabolomic studies have employed nuclear magnetic resonance (NMR) spectrometry to identify disease-specific metabolites related to fat metabolism and intestinal microbial metabolism. A second major unmet need exists for biomarkers of disease activity that have superiority over standard C-reactive protein assessment and reflect MRI inflammation in the axial spine. Several biomarkers reflecting inflammation (calprotectin), angiogenesis (vasoactive endothelial growth factor), and connective tissue turnover (C2M, C3M, and citrullinated metalloproteinase degraded fragment of vimentin) have recently been shown to reflect disease activity when compared with clinical outcomes but comparisons with MRI inflammation are very limited. With increasing availability of highly effective but costly therapies, a third unmet need is biomarkers that can predict response to therapies with different mechanisms of action and are superior to C-reactive protein. Calprotectin is currently the only candidate. Although there are as yet no proven therapies for preventing progression of disease there is an unmet need for biomarkers of prognosis that are more responsive than radiography. Aside from CRP no consistent candidates have emerged. Future studies will need to be prospective, include consecutive patients presenting with undiagnosed back pain, and use more reliable and objective endpoints such as MRI inflammation. Moreover, it has become evident that targeted biomarker studies have not been successful in identifying clinically useful biomarkers and technologies that can simultaneously assess “multiomic” markers will need to be analyzed for future advances. These include more sophisticated metabolomic profiling and universal metabolome-standard (UMS) methodology, next generation RNA sequencing, and affinity-based quantitative proteomics based on the use of nucleic acid binders such as the aptamer-based SOMAscan assay.
Species diversity and seasonal abundance of Culicoides biting midges in northwestern Argentina

The species diversity and seasonal abundance of biting midges of the genus Culicoides (Diptera: Ceratopogonidae) were studied in northwestern Argentina during the period 2003–2005. A total of 5437 Culicoides specimens were collected using CDC light traps in three areas of the mountainous rainforest area. The most common species were Culicoides paraensis (Goeldi) and C. insignis Lutz, Culicoides lahillei (Iches), C. venezuelensis Ortiz & Mirsa, C. debilipalpis Lutz and C. crescentis Wirth & Blanton were also collected. Culicoides paraensis was abundant during the summer, and C. insignis and C. lahillei during late summer and early fall. Accumulated rainfall was the climatic variable most related to fluctuation in abundance of C. paraensis.
Direct Effects of Retinoic Acid on Entry of Fetal Male Germ Cells into Meiosis in Mice1

In both male and female germ cells of mice, retinoic acid (RA) is a meiosis-inducing factor. In the present study, we used a germ cell culture system to examine the direct effects of RA on meiotic initiation in male germ cells at the stage when they normally enter mitotic arrest to determine the extent to which fetal male germ cells can respond to exogenous RA to alter their sex-specific pathway. Male germ cells between 13.5 and 15.5 days postcoitum (dpc) were isolated from Pou5fl-green fluorescent protein transgenic fetuses and cultured with or without RA for up to 6 days. In the absence of RA, male germ cells did not undergo DNA replication and did not enter meiosis in culture. However, in the presence of RA, male germ cells isolated at 13.5 dpc expressed Stra8 and initiated the meiotic process. The ratio of cells entering meiosis gradually decreased as cells were isolated progressively at later stages. By 15.5 dpc, isolated male germ cells lost their ability to respond to RA signaling. These cells remained dispersed as single cells and progressed along the male differentiation pathway, as evidenced by the establishment of male-specific methylation imprints regardless of the presence or absence of RA. We conclude that male germ cells maintain sexual bipotency until 14.5 dpc that can be reversed by the addition of RA. Once male germ cells enter mitotic arrest, however, they appear to be committed irreversibly to the male-specific differentiation pathway even in the presence of exogenously added RA.
Risk for the introduction of exotic ticks and pathogens into Italy through the illegal importation of tortoises, Testudo graeca

In April 2008, 585 tortoises illegally imported into Italy from North Africa were examined for the presence of ticks. Of these, 221 tortoises (37.8%) were infested with a mean intensity of 3.9 ± 3.1 ticks (range 1–17 ticks). A total of 798 ticks (672 males, 125 females and one nymph) were collected and identified as Hyalomma aegyptium (L.) (Acari: Ixodidae). The overall male : female ratio observed was 5 : 1. The prevalence and mean intensity [±standard deviation (SD)] of ticks were higher among male (67.4%, 4.0 ± 3.2) than female (55.6%, 3.8 ± 3.1) tortoises, although this difference was not significant. By contrast, the prevalence and mean intensity of ticks were significantly higher on tortoises weighing >100 g (61.5%, 4.0 ± 3.2) compared with tortoises weighing <100 g (12.1%, 2.1 ± 1.1). Of the infested tortoises, 89.8% had ticks on their hind limbs, 21.0% on forelimbs, 18.6% on the tail and pre‐anal area, and 4.8% on the head; thus the hind limbs were evidently the preferred attachment site. The present report highlights the need to develop surveillance systems to prevent the introduction and spreading of exotic ticks and tick‐borne pathogens in Italy and other European countries.
Incrimination of the mosquito, Aedes taeniorhynchus, as the primary vector of heartworm, Dirofilaria immitis, in coastal Yucatan, Mexico

Mosquito collections were carried out on microfilaraemic dogs, positive for Dirofilaria sp. , for 18 consecutive nights in the coastal town of Celestún, Yucatan, southeast Mexico, during the rainy season (August) of 2007. A total of 292 female mosquitoes representing 12 species of dipteran Culicidae were collected: Anopheles albimanus (Wiedemann); Anopheles crucians (Wiedemann); Anopheles pseudopunctipennis (Theobald); Culex coronator (Dyar & Knab); Culex interrogator (Dyar & Knab); Culex nigripalpus (Theobald); Culex quinquefasciatus (Say); Culex salinarius (Coquillett); Aedes aegypti (Linnaeus); Aedes scapularis (Rondani); Aedes sollicitans (Walker), and Aedes taeniorhynchus (Wiedemann). Aedes taeniorhynchus and Cx. quinquefasciatus were the species found most commonly feeding on the dogs. Filarial nematodes were observed by microscopy in nine of the mosquito species collected; however, third‐instar larvae were only observed in Ae. taeniorhynchus and An. crucians. Of 76 Ae. taeniorhynchus specimens found positive for Dirofilaria sp. by dissection, 14 were confirmed to be positive for Dirofilaria immitis (Leidy) by polymerase chain reaction (PCR). The resulting infection rate for D. immitis confirmed by PCR (6.2%) is higher than any infection rate for Ae. taeniorhynchus previously reported from the Americas.
Responses of tabanids to Nzi traps baited with octenol, cow urine and phenols in Canada

Cow urine and the two phenols responsible for the attraction of biting flies to cow urine (4‐methylphenol, 3‐n‐propylphenol) were compared with octenol (1‐octen‐3‐ol) as baits for Tabanidae. Relative to an unbaited Nzi trap, catches of the horseflies Hybomitra lasiophthalma (Macquart), Tabanus similis Macquart and Tabanus quinquevittatus Wiedemann (Diptera: Tabanidae) were increased by 1.5–2.6, 1.4–2.0 and 1.4–1.9 times, respectively, whenever a bait included octenol released at either 0.13 mg/h or 1.5 mg/h, regardless of the presence of phenols or urine. Catches were not affected when traps were baited with phenols alone at evaporation rates of 0.38 mg/h (4‐methylphenol) and 0.022 mg/h (3‐n‐propylphenol). Catches of Hybomitra horseflies were increased by 1.5–1.9 times with cow urine and 2.6 times with cow urine + octenol. This bait combination could prove to be particularly useful for Hybomitra horseflies, the common tabanids of northern environments.
Regulation of RIPK3- and RHIM-dependent Necroptosis by the Proteasome*

Receptor-interacting protein kinase 3 (RIPK3) is a serine/threonine kinase with essential function in necroptosis. The activity of RIPK3 is controlled by phosphorylation. Once activated, RIPK3 phosphorylates and activates the downstream effector mixed lineage kinase domain-like (MLKL) to induce necroptosis. In certain situations, RIPK3 has also been shown to promote apoptosis or cytokine expression in a necroptosis and kinase-independent manner. The ubiquitin-proteasome system is the major pathway for selective degradation of cellular proteins and thus has a critical role in many cellular processes such as cell survival and cell death. Clinically, proteasome inhibition has shown promise as an anti-cancer agent. Here we show that the proteasome inhibitors MG132 and bortezomib activate the RIPK3-MLKL necroptotic pathway in mouse fibroblasts as well as human leukemia cells. Unlike necroptosis induced by classical TNF-like cytokines, necroptosis induced by proteasome inhibitors does not require caspase inhibition. However, an intact RIP homotypic interaction motif (RHIM) is essential. Surprisingly, when recruitment of MLKL to RIPK3 is restricted, proteasome inhibitors induced RIPK3-dependent apoptosis. Proteasome inhibition led to accumulation of K48-linked ubiquitinated RIPK3, which was partially reduced when Lys-264 was mutated. Taken together, these results reveal the ubiquitin-proteasome system as a novel regulatory mechanism for RIPK3-dependent necroptosis.