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Surgical sterilisation is a common tool in most of the developed world for combatting the overpopulation of unwanted dogs and eliminating the risk of reproductive diseases in pet dogs. Luteinising hormone (LH) stimulates the secretion of gonadal steroid hormones, which negatively feedback to the hypothalamus and anterior pituitary to decrease the secretion of GnRH and LH, respectively. However, in the desexed mammal, there is no negative feedback of LH, which results in supraphysiological circulating concentrations. LH receptors in non-reproductive tissues may induce cell division and stimulate nitric oxide release. With constant activation following gonad removal, these receptors are upregulated, further magnifying the effects of the supraphysiological LH concentrations in non-reproductive tissues. Non-reproductive long-term health complications resulting from canine desexing are discussed, as well as how these effects are mediated by LH receptor activation in non-reproductive target tissues. [ABSTRACT FROM AUTHOR]

Contents Overpopulation of cats and dogs is a serious worldwide problem that demands novel, safe and cost-effective solutions. The objective of this study was to generate and characterize phage-peptide conjugates with gonadotropin-releasing hormone ( Gn RH) for potential use as an immunocontraceptive. A filamentous phage vector f5-8 with wild-type phage coat proteins was used as a carrier for construction of chemical conjugates with Gn RH, a peptide that acts as a master reproductive hormone. In such conjugates, the phage body plays the role of a carrier protein, while multiple copies of Gn RH peptide stimulate production of neutralizing anti- Gn RH antibodies potentially leading to contraceptive effects. To generate the constructs, four different Gn RH-based peptides were synthesized and conjugated to phage particles in a two-step procedure: (i) peptides were reacted with phage to form a conjugate using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride chemistry ( EDC) and (ii) the conjugates were separated from remaining free peptides by dialysis. Formation and specificity of phage- Gn RH conjugates were confirmed by three independent methods: spectrophotometry, electron microscopy and ELISA. When the conjugates were tested for interaction with sera collected from cats and dogs immunized with Gn RH-based vaccines in independent studies, strong specific ELISA signals were obtained, suggesting the potential use of the conjugates for cat and dog immunosterilization. The ability of the conjugates to stimulate production of anti- Gn RH antibodies in vivo was evaluated in mice. While optimization of dose, immunization route and adjuvant still requires investigation, our preliminary results demonstrated the presence of anti- Gn RH antibodies in sera of mice immunized with such conjugates. Fertility trials in cats and dogs will be needed to evaluate contraceptive potentials of the phage- Gn RH peptide chemical conjugates. [ABSTRACT FROM AUTHOR]

Contents Four German Shorthair Pointer bitches each produced from two to five (total of 14) purebred litters in response to natural matings to either natural oestrus (n = 8) or oestrus induced by Ovuplant®, a sustained-release implant containing 2.1 mg deslorelin (n = 6). All bitches initially produced litters from natural oestrous matings prior to producing litters from induced oestrus, and two of the bitches also produced natural litters subsequent to their Ovuplant® litters. Mean litter size was lower for Ovuplant® litters (5.4 vs 8.6 puppies; p < 0.001) and within each bitch every induced litter was smaller than any of her natural litters. [ABSTRACT FROM AUTHOR]

Two retrogene insertions, FGF4L1 (formerly 18-FGF4, colloquially CDPA) and FGF4L2 (formerly 12-FGF4, colloquially CDDY), have recently been discovered as determinants of short leg phenotype in dogs. This case study is comprised of a family of standard wirehaired dachshunds in which the dogs lacking the FGF4L1 gene exhibit a tall phenotype. The tall phenotype in the dachshunds of this report precludes the dog's working function of den work. The data presented in this report provide information as to how FGF4L1 status could be used in making breeding decisions in dachshunds to maintain working ability without compromising animal health., Competing Interests: Each of the authors breeds and exhibits dachshunds. KS has authored three books on working dachshunds and is currently writing a fourth., (Copyright © 2025 Sullivan, Szeremeta and Kutzler.)

Background: The relationship between radiographic disc calcification score and FGF4L2 genotype has been reported in only a small number of dachshunds., Hypothesis: A genotype with either 0 or 1 FGF4L2 copy will be associated with a lower number of calcified discs (lower K-n) compared with a genotype with 2 FGF4L2 copies., Animals: Dachshunds registered with the Norwegian or Finnish Kennel Clubs for which both K-n and FGF4L2 genotype are known (n = 407)., Methods: Retrospective study (2012-2024). The frequency and percentage of dachshunds within each K-n group (K0, K1/2, K3/4, K5+) by FGF4L2 genotype (FGF4L2/FGF4L2, N/FGF4L2, N/N) were reported. The K-n distribution differences among genotypes were analyzed by the Cochran-Mantel-Haenszel test in SAS 9.4. Significance was defined as P < .05., Results: A difference in K-n distribution was found between dachshunds that have 0 or 2 FGF4L2 copies. However, no difference was found in the K-n distribution between dachshunds that have 1 or 2 FGF4L2 copies., Conclusions and Clinical Importance: Most dachshunds with 1 FGF4L2 copy have radiographic disc calcification scores that are associated with substantially lower risk of symptomatic disc disease. Radiographic disc calcification scores are similar in dachshunds with 0 and 1 FGF4L2 copies. Given the high allele frequency of FGF4L2, breeding to produce progeny with 1 FGF4L2 copy is expected to be a more achievable short-term goal for dachshund spinal health breeding programs than breeding for 0 copies. It is recommended that both K-n and FGF4L2 genotype status be considered when choosing dachshund breeding stock., (© 2024 The Author(s). Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

The potential association of milk with childhood obesity has been widely debated and researched. Milk is known to contain many bioactive compounds as well as bovine exosomes rich in micro-RNA (miR) that can have effects on various cells, including stem cells. Among them, adipose stem cells (ASC) are particularly interesting due to their role in adipose tissue growth and, thus, obesity. The objective of this study was to evaluate the effect of milk consumption on miR present in circulating exosomes and the transcriptome of ASC in piglets. Piglets were supplemented for 11 weeks with 750 mL of whole milk (n = 6; M) or an isocaloric maltodextrin solution (n = 6; C). After euthanasia, ASC were isolated, quantified, and characterized. RNA was extracted from passage 1 ASC and sequenced. Exosomes were isolated and quantified from the milk and plasma of the pigs at 6-8 hours after milk consumption, and miRs were isolated from exosomes and sequenced. The transfer of exosomes from milk to porcine plasma was assessed by measuring bovine milk-specific miRs and mRNA in exosomes isolated from the plasma of 3 piglets during the first 6h after milk consumption. We observed a higher proportion of exosomes in the 80 nM diameter, enriched in milk, in M vs. C pigs. Over 500 genes were differentially expressed (DEG) in ASC isolated from M vs. C pigs. Bioinformatic analysis of DEG indicated an inhibition of the immune, neuronal, and endocrine systems and insulin-related pathways in ASC of milk-fed pigs compared with maltodextrin-fed pigs. Of the 900 identified miRs in porcine plasma exosomes, only 3 miRs were differentially abundant between the two groups and could target genes associated with neuronal functions. We could not detect exosomal miRs or mRNA transfer from milk to porcine-circulating plasma exosomes. Our data highlights the significant nutrigenomic role of milk consumption on ASC, a finding that does not appear to be attributed to miRs in bovine milk exosomes. The downregulation of insulin resistance and inflammatory-related pathways in the ASC of milk-fed pigs should be further explored in relation to milk and human health. In conclusion, the bioinformatic analyses and the absence of bovine exosomal miRs in porcine plasma suggest that miRs are not vertically transferred from milk exosomes., Competing Interests: The authors have declared that no competing interests exist, (Copyright: © 2024 Swanson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: The expression of luteinizing hormone (LH) receptors has been reported in canine nonneoplastic and neoplastic lymphocytes. This study aimed to determine the effect of LH receptor activation in neoplastic canine T lymphocytes., Methods: Cell lines (CLC, CLK, EMA) derived from 3 dogs with primary T-cell lymphoma were used. Cell lines were previously phenotyped and evaluated for LH receptor expression with flow cytometry. Cell lines were stimulated with increasing concentrations (0, 4, 400, and 4,000 U/mL) of human chorionic gonadotropin (hCG; an LH receptor agonist), RNA was extracted, cDNA was synthesized, and gene expression was determined using quantitative PCR and the 2-ΔΔCt method. Cell lines were stimulated with the same increasing concentrations of hCG, and cell proliferation, adhesion, and transmigration were determined using commercial assays. The effects of LH receptor activation were compared between hCG concentrations using a one-way ANOVA. Significance was defined as P < .05., Results: LH receptor stimulation increased LH receptor gene expression in the CLC cell line, and there was a trend for increased expression in the EMA cell line but no effect in the CLK cell line. Activation of LH receptors increased proliferation in all 3 cell lines, endothelium adhesion in 2 cell lines (CLC, CLK), and transmigration in 2 cell lines (CLK, EMA)., Conclusions: LH receptor stimulation using hCG increased LH receptor gene expression in neoplastic canine T lymphocytes and increased cell proliferation, adhesion, and transmigration., Clinical Relevance: These findings may provide a physiologic mechanism for the increased incidence of lymphoma reported in dogs with sustained supraphysiologic LH concentrations.

Introduction: Distal femur fractures account for 3-6% of all femur fractures. Internal fixation of most distal femur fractures with an anatomic lateral locking plate should permit some motion at the metaphyseal portion of the fracture when secondary bone healing is planned by the operating surgeon. While several studies have been performed evaluating union rates for distal femur fractures with stainless steel and titanium plates, the timing of callus formation between stainless steel and titanium implants used as bridge plates for distal femur fractures (AO/OTA 33-A and -C) has been investigated to a lesser extent. We hypothesize that callus will be visualized earlier with post-operative radiographs with titanium versus stainless steel bridge plates., Methods: We retrospectively reviewed a consecutive cohort of patients over 18 years of age with acute AO/OTA 33-A and 33-C fracture patterns treated with an isolated stainless steel or titanium lateral bridge plate within 4 weeks of injury by a single fellowship-trained orthopedic trauma surgeon from 2011 to 2020 at one academic Level 1 trauma center. An independent, fellowship-trained orthopedic trauma attending surgeon reviewed anterior-posterior (AP) and lateral radiographs from every available post-operative clinic visit and graded them using the Modified Radiographic Score for Tibia (mRUST)., Results: Twenty-five subjects were included in the study with 10 with stainless steel and 15 with titanium plates. There were no significant differences in demographics between both groups, including age, sex, BMI, injury classification, open versus closed, mechanism, and laterality. Statistically significant increased mRUST scores, indicating increased callus formation, were seen on 12-week radiographs (8.4 vs. 11.9, p = 0.02) when titanium bridge plates were used. There were no statistically significant differences in mRUST scores at 6 or 24-weeks, but scores in the titanium group were higher in at every timepoint., Discussion: In conclusion, we observed greater callus formation at 12 weeks after internal fixation of 33-A and 33-C distal femur fractures treated with titanium locked lateral distal femoral bridge plates compared to stainless steel plates. Our data suggest that titanium metallurgy may have quicker callus formation compared to stainless steel if an isolated, lateral locked bridge plate is chosen for distal femur fracture fixation., (© 2024. The Author(s), under exclusive licence to Springer-Verlag France SAS, part of Springer Nature.)

Introduction: Predictive models for Clostridioides difficile infection can identify high-risk patients and aid clinicians in preventing infection. Issues of generalizability regarding current predictive models have been acknowledged but, to the authors' knowledge, have never been quantified., Methods: C. difficile infection, severity and recurrence predictive models were created using multi-variate logistic regression through case-control sampling from an urban safety-net hospital. Models were validated using five-fold cross-validation, and inverse probability weights (IPW) based on two different catchment area definitions were used to improve external validity. Akaike Information Criterion (AIC), area under the receiver operating characteristic curve (AUROC), and sensitivity and specificity with bootstrapped confidence intervals (CI) were used to assess and compare model fit and performance., Results: Changes in performance before and after weighting were small across all models, although differences were more apparent after weighting the recurrence model (AUROC values of 0.78, 0.76 and 0.71 for the unweighted and two weighted models, respectively). Overall, the infection model performed the best (AUROC 0.82, 95% CI 0.78-0.85), followed by the recurrence model (AUROC 0.78, 95% CI 0.69-0.86) and then the severity model (AUROC 0.70, 95% CI 0.63-0.78)., Conclusions: The performance of the models after weighting did not change drastically, suggesting that the models predicting C. difficile infection, severity and recurrence may not be impacted by patient selection factors. However, other researchers may wish to consider addressing these catchment forces using IPW., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The purpose of this research was to quantify sperm acrosome associated 3 protein expression in the ovaries of young (3.0 ± 0.9 months, n = 11) and adult (10.4 ± 2.8 months, n = 11) queens. Immunohistochemistry was performed on formalin-fixed, paraffin-embedded feline ovarian sections. Ovaries were obtained following routine ovariohysterectomy of queens. Cellular expression of sperm acrosome associated 3 protein was measured in primordial, primary, secondary, and tertiary follicles using an image-analysis software's red, green, and blue stack and manual thresholding functions. The oocyte nucleus, ooplasm, granulosa cells, and theca cells were outlined using the freehand selection tool and mean grey value was recorded. Results from each cellular location were compared between age groups using a Student's t-test and between follicle stages using an analysis of variance. Compared to adult queens, younger queens had significantly greater sperm acrosome associated 3 protein expression in granulosa cells of primary, secondary, and tertiary follicles. Also, theca cells of secondary and tertiary follicles had significantly greater sperm acrosome associated 3 protein expression in younger queens compared to adult queens. The oocyte nucleus of primordial, primary, and secondary follicles had significantly greater sperm acrosome associated 3 protein expression in younger queens compared to adult queens. However, sperm acrosome associated 3 protein expression within the ooplasm did not differ significantly between age groups of any follicle type. More research is needed to determine what role sperm acrosome associated 3 protein may play in female fertility in animals as well as what mechanisms regulate ovarian sperm acrosome associated 3 protein expression over time., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Despite the overwhelming success of mRNA-based vaccine in protecting against SARS-CoV-2 infection and reducing disease severity and hospitalization, little is known about the role lipid nanoparticles (LNP) play in initiating immune response. In this report we studied the adjuvantive impact of empty LNP with no mRNA cargo (eLNP) on anti-viral pathways and immune function of cells from young and aged individuals. We found that eLNP induced maturation of monocyte derived dendritic cells by measuring the expression of CD40, CD80, HLA-DR and production of cytokines including IFN-α,IL-6, IFN-γ, IL-12, and IL-21. Flow cytometry analysis of specific dendritic cell subsets showed that eLNP can induce CD40 expression and cytokine production in cDC1, cDC2 and monocytes. Empty LNP (eLNP) effects on dendritic cells and monocytes coincided with induction pIRF7 and pTBK1, which are both important in mitigating innate immune signaling. Interestingly our data show that in response to eLNP stimulus at 6 and 24 hrs, aged individuals have decreased CD40 expression and reduced IFN- γ output compared to young adults. Furthermore, we show that cDC1, cDC2, and CD14 dim CD16 + monocytes from healthy aged individuals have dysregulated anti-viral signaling response to eLNP stimulation as measured by the defect in type I IFN production, phosphorylation of IRF7, TBK-1, and immune function like phagocytosis. These data showed a novel function of eLNP in eliciting DC maturation and innate immune signaling pathways and that some of these functions are impaired in older individuals providing some suggestion of why older individuals (> 65 yrs of age) respond display lower immune responses and adverse events to SARS-CoV-2 mRNA-based vaccines.

Luteinizing hormone receptors (LHRs) are expressed in canine lymphoma and hemangiosarcoma. We hypothesized that LHR would be expressed in canine mast cell tumors (MCTs) and that more neoplastic mast cells would express LHR in gonadectomized dogs compared with intact dogs. Eleven archived formalin-fixed paraffin-embedded cutaneous MCT tissue sections were processed using routine immunohistochemistry. For both the KIT protein and LHR, the percentage of positive cells for each staining pattern (I-III) was calculated. A Student's t test was used to compare the total percentage of positive cells expressing LHR and KIT in intact and gonadectomized dogs. A one-way analysis of variance was used to compare the percentage of cells within each staining pattern for LHR and KIT in intact and gonadectomized dogs. All MCT expressed LHR. MCT from gonadectomized dogs had a significantly higher percentage of LHR-positive mast cells (84.2 ± 8.7%) compared with MCTs from intact dogs (64.3 ± 4.2%). This is the first study to demonstrate the expression of LHR in canine MCTs and to report that LHR expression is increased in neoplastic mast cells from gonadectomized dogs compared with intact dogs. Future studies are planned to evaluate the functionality of the LHR in canine neoplastic mast cells., (© 2022 by American Animal Hospital Association.)

KR13, a peptide triazole thiol previously established to inhibit HIV-1 infection and cause virus lysis, was evaluated by flow cytometry against JRFL Env-presenting cells to characterize induced Env and membrane transformations leading to irreversible inactivation. Transiently transfected HEK293T cells were preloaded with calcein dye, treated with KR13 or its thiol-blocked analogue KR13b, fixed, and stained for gp120 (35O22), MPER (10E8), 6-helix-bundle (NC-1), immunodominant loop (50-69), and fusion peptide (VRC34.01). KR13 induced dose-dependent transformations of Env and membrane characterized by transient poration, MPER exposure, and 6-helix-bundle formation (analogous to native fusion events), but also reduced immunodominant loop and fusion peptide exposure. Using a fusion peptide mutant (V504E), we found that KR13 transformation does not require functional fusion peptide for poration. In contrast, simultaneous treatment with fusion inhibitor T20 alongside KR13 prevented membrane poration and MPER exposure, showing that these events require 6-helix-bundle formation. Based on these results, we formulated a model for PTT-induced Env transformation portraying how, in the absence of CD4/co-receptor signaling, PTT may provide alternate means of perturbing the metastable Env-membrane complex, and inducing fusion-like transformation. In turn, the results show that such transformations are intrinsic to Env and can be diverted for irreversible inactivation of the protein complex.

Estrus synchronization and manipulation are a tool that has been used by producers to provide uniform lamb and kid meat production and dairy sheep and goat milk production, to concentrate work and labor cost, and to plan for the lambing and kidding time. Breeders can also use estrus synchronization to stimulate ewes and does to exhibit estrus and ovulate outside of the breeding season, although both the ovulation rate and pregnancy rate may be decreased. To increase the ovulation rate outside of the breeding season, a variety of estrus synchronization methods have been used., Competing Interests: Disclosure The author has nothing to disclose., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Neonatal maladjustment syndrome is characterized by depressed consciousness, neurologic signs, and reduced or nonexistent suckle reflex. Resuscitation compression (squeezing) has been used in newborn foals and calves to reduce the behavioral symptoms of neonatal maladjustment syndrome. In this review, the authors describe how resuscitation compression can be used in newborn sheep. This technique can improve abnormal neonatal lamb behavior and encourage suckling in resistant lambs., Competing Interests: Disclosure This research was funded by the Oregon Sheep Commission (USA) and the College of Agricultural Sciences, Oregon State University, Continuing Researchers Support Program (USA)., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Hysteroscopy in alpacas and llamas allows for the identification of abnormalities on the surface or within the endometrium that cannot be identified with other methods. Hysteroscopy also allows for site-directed endometrial cytology, culture, and biopsy to achieve a definitive diagnosis. Even when no cause for infertility can be found, previously infertile females tend to become pregnant and maintain their pregnancies to term following the hysteroscopic procedure. This therapeutic effect may be a response to pre-hysteroscopy estrogen treatment, dilation of the uterine horns during hysteroscopy, and/or posttreatment uterine lavages. Complications following hysteroscopy have not been reported in camelids., Competing Interests: Disclosure The authors have nothing to disclose., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The SARS-CoV-2 pandemic has almost 56 million confirmed cases resulting in over 1.3 million deaths as of November 2020. This infection has proved more deadly to older adults (those >65 years of age) and those with immunocompromising conditions. The worldwide population aged 65 years and older is increasing, and the total number of aged individuals will outnumber those younger than 65 years by the year 2050. Aging is associated with a decline in immune function and chronic activation of inflammation that contributes to enhanced viral susceptibility and reduced responses to vaccination. Here we briefly review the pathogenicity of the virus, epidemiology and clinical response, and the underlying mechanisms of human aging in improving vaccination. We review current methods to improve vaccination in the older adults using novel vaccine platforms and adjuvant systems. We conclude by summarizing the existing clinical trials for a SARS-CoV-2 vaccine and discussing how to address the unique challenges for vaccine development presented with an aging immune system.

The Apgar (appearance, pulse, grimace, attitude, respiration) score is an accepted method for assessing newborn viability in humans and domestic animals. The purpose of this research was to modify an existing Apgar scoring system for lambs and compare Apgar score with litter size and birth order, as well as time-to-stand and time-to-suckle. Polypay lambs (n = 75) delivered naturally were used for this study. Pulse (<100 bpm = 0; 100-175 bpm = 1; >175 bpm = 2) and respiration (<45% SpO 2  = 0; 45-65% SpO 2  = 1; >65% SpO 2  = 2) were measured with a pulse oximeter attached to the lamb's ear. Appearance, grimace, and attitude were assessed using mucous membrane color (cyanotic = 0; pale = 1; pink = 2); response to nose stimulation (no response = 0; moves head slightly = 1; shakes head = 2); and response to rump stimulation (no movement = 0; moves but no attempt to stand = 1; attempts to stand = 2), respectively. The Apgar score was measured within 5 min of birth (T0), 15 min (T15) and 60 min (T60) after birth. The time-to-stand and time-to-suckle were recorded and compared to Apgar scores. Apgar scores did not differ by litter size or birth order. There were no significant correlations between the Apgar scores and time-to-stand. There was a moderate positive correlation between attitude and time-to-suckle at T0 (R 2  = 0.467), which may affect the ingestion of adequate colostrum in a timely manner. Producers may choose to consider using Apgar scoring so that assistance can be provided to lambs before serious complications arise., (Copyright © 2020 Elsevier Inc. All rights reserved.)

An effective prophylactic vaccine targeting HIV must induce a robust humoral response and must direct the bulk of this response to the mucosa-the primary site of HIV transmission. The chemokine, CCL28, is secreted by epithelial cells at mucosal surfaces and recruits' cells expressing its receptor CCR10. CCR10 is predominantly expressed by IgA + ASCs. We hypothesized that co-immunization with plasmid DNA encoding consensus envelope antigens with plasmid-encoded CCL28 would enhance anti-HIV IgA responses at mucosal surfaces. Indeed, animals receiving pCCL28 and pEnvA/C had significantly increased HIV-specific IgA in fecal extract. Surprisingly, CCL28 co-immunization induced a significant increase in anti-HIV IgG in the serum in mice compared to those receiving pEnvA/C alone. These robust antibody responses were not associated with changes in the frequency of germinal center B cells but depended upon the expression of CCR10, as these responses we abolished in CCR10-deficient animals. Finally, immunization with CCL28 led to increased frequencies in HIV-specific CCR10 + and CCR10 + IgA + B cells in the small intestine and Peyer's patches of vaccinated animals as compared to those receiving pEnvA/C alone. These data indicate that CCL28 administration can enhance antigen-specific humoral responses systemically and at mucosal surfaces., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier Ltd.)

Dual-acting virucidal entry inhibitors (DAVEIs) have previously been shown to cause irreversible inactivation of HIV-1 Env-presenting pseudovirus by lytic membrane transformation. This study examined whether this transformation could be generalized to include membranes of Env-presenting cells. Flow cytometry was used to analyze HEK293T cells transiently transfected with increasing amounts of DNA encoding JRFL Env, loaded with calcein dye, and treated with serial dilutions of microvirin (Q831K/M83R)-DAVEI. Comparing calcein retention against intact Env expression (via Ab 35O22) on individual cells revealed effects proportional to Env expression. "Low-Env" cells experienced transient poration and calcein leakage, while "high-Env" cells were killed. The cell-killing effect was confirmed with an independent mitochondrial activity-based cell viability assay, showing dose-dependent cytotoxicity in response to DAVEI treatment. Transfection with increasing quantities of Env DNA showed further shifts toward "High-Env" expression and cytotoxicity, further reinforcing the Env dependence of the observed effect. Controls with unlinked DAVEI components showed no effect on calcein leakage or cell viability, confirming a requirement for covalently linked DAVEI compounds to achieve Env transformation. These data demonstrate that the metastability of Env is an intrinsic property of the transmembrane protein complex and can be perturbed to cause membrane disruption in both virus and cell contexts.

MicroRNAs are conserved, endogenous small RNAs with critical post-transcriptional regulatory functions throughout eukaryota, including prominent roles in development and disease. Despite much effort, microRNA annotations still contain errors and are incomplete due especially to challenges related to identifying valid miRs that have small numbers of reads, to properly locating hairpin precursors and to balancing precision and recall. Here, we present miRWoods, which solves these challenges using a duplex-focused precursor detection method and stacked random forests with specialized layers to detect mature and precursor microRNAs, and has been tuned to optimize the harmonic mean of precision and recall. We trained and tuned our discovery pipeline on data sets from the well-annotated human genome, and evaluated its performance on data from mouse. Compared to existing approaches, miRWoods better identifies precursor spans, and can balance sensitivity and specificity for an overall greater prediction accuracy, recalling an average of 10% more annotated microRNAs, and correctly predicts substantially more microRNAs with only one read. We apply this method to the under-annotated genomes of Felis catus (domestic cat) and Bos taurus (cow). We identified hundreds of novel microRNAs in small RNA sequencing data sets from muscle and skin from cat, from 10 tissues from cow and also from human and mouse cells. Our novel predictions include a microRNA in an intron of tyrosine kinase 2 (TYK2) that is present in both cat and cow, as well as a family of mirtrons with two instances in the human genome. Our predictions support a more expanded miR-2284 family in the bovine genome, a larger mir-548 family in the human genome, and a larger let-7 family in the feline genome., Competing Interests: The authors have declared that no competing interests exist.

The objectives of this study were to determine the effects of acupuncture in dairy cows (Bos taurus) on caruncular matrix metalloproteinase type-2 (MMP2) at 0, 2 and 4 hr after calving. Acupuncture (n = 6) was applied at 0 and 2 hr after calving to 6 points that relax the cervix and stimulate uterine contractions. Controls (n = 9) were kept in a stanchion for 15 min without acupuncture. All of the cows in the study delivered their placenta in <4 hr. Formalin-fixed caruncles were paraffin-embedded and subjected to routine immunohistochemistry to determine MMP2 expression, which was scored by a single observer. Flash frozen caruncles were homogenized, and protein concentration was determined. MMP2 concentrations were calculated using commercial bovine ELISAs. MMP2 enzyme activity was determined using zymography. The mean value for each time point for each cow was used to calculate the mean ± SEM for each treatment group. MMP2 was predominantly localized to the epithelial and subepithelial stromal cells of the caruncles in both treatment groups. MMP2 immunoexpression was lower 4 hr after calving in the control cows (p = 0.012) but not in the acupuncture treated cows indicating that acupuncture treatment maintained MMP expression. MMP2 tissue concentration was lower 2 hr after calving in the control cows (p = 0.048) but not in the acupuncture treated cows. MMP2 enzyme activity decreased from 0 to 2 hr after calving in control cows (p = 0.046) but not in acupuncture treated cows. This study provides physiologic evidence for the effects of acupuncture on the bovine reproductive tract and substantiates the use of this treatment in cases of placental retention., (© 2019 Blackwell Verlag GmbH.)

The effect of milk consumption on childhood obesity is unclear and a direct demonstration of an association is needed. In the present study, we used piglets as a model for prepubertal children to determine the effect of milk on adipose tissue. Two studies were conducted: study 1 with 5-wk-old male piglets (n = 8) and study 2 with 8- to 9-wk-old male piglets (n = 12). The piglets were fed a normal growing diet and randomly assigned to receive daily either 750 mL of whole cow milk or an isocaloric maltodextrin solution (control). For approximately 12 wk, body weight, feed intake, and subcutaneous back fat thickness were determined ultrasonographically and recorded. At euthanasia, back and neck fat thicknesses were measured and samples of back fat were collected for adipose histology. In study 1, but not study 2, piglets receiving milk grew more and ate more compared with control. In study 1, both back fat and neck fat thickness were greater in the milk-fed piglets and they had a higher frequency of small adipocytes and a lower frequency of intermediate and large adipocytes compared with controls. In study 2, control pigs had a significantly greater frequency of intermediate adipocytes but the milk-fed piglets tended to have a higher frequency of the largest adipocytes. In conclusion, milk has no apparent causal or consistent effect on adipose tissue in growing piglets., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

It is now widely accepted in human medicine that prolactin (PRL) and growth hormone (GH) function in the mammary gland in an autocrine and paracrine manner in tumour formation. The aim of this study was to compare PRL and GH immunoactivity in canine mammary tumours submitted for histopathologic evaluation. Formalin-fixed specimens from spontaneously occurring mammary adenomas and adenocarcinomas from 24 female client-owned dogs were used. Information pertaining to the reproductive status of the patient at the time of mammary tumour diagnosis was obtained from each of the submitting veterinarians. Tissues were paraffin-embedded and sectioned (5 μm) onto charged slides. All slides were deparaffinized and rehydrated. Endogenous peroxidase activity was inactivated with 3% H 2 O 2, and non-specific binding was blocked. Polyclonal rabbit antihuman PRL (DAKO A0569) and GH antibody (DAKO A0570) were applied at a 1:250 and 1:200 dilutions, respectively. A universal rabbit negative control (DAKO N1699) was used. Slides were then reacted with anti-rabbit horseradish peroxidase followed by Nova Red Peroxidase substrate. Slides were counter-stained with haematoxylin, dehydrated and mounted. Tumour type and reproductive status at time of tumour diagnosis were compared individually between tumours that were negative or positive for PRL and GH using a two-tailed analysis of variance. Significance was defined as p < .05. There was no significant relationship between tumour type and PRL and GH presence. In addition, reproductive status at the time of tumour removal was found to be not significant. These results vary from previous reports in canine mammary tumours and warrant further investigation., (© 2016 Blackwell Verlag GmbH.)

Sterilization is a key strategy to reduce the number of domestic cats entering and killed in shelters each year. However, surgical sterilization is expensive and labour-intensive and cannot fully address the 70 million free-roaming cats estimated to exist in the United States. GonaCon™ is a gonadotropin-releasing hormone vaccine originally developed for use as a wildlife immunocontraceptive. An earlier formulation was tested in domestic cats and found to be safe and effective for long-term contraception. However, the current Environmental Protection Agency (EPA)-registered formulation consists of a different antigen-carrier protein and increased antigen concentration and has never been tested in cats. A pilot study was undertaken to evaluate the short-term safety of a single GonaCon immunization, assess the consequences of vaccinated cats receiving an accidental second GonaCon injection and determine the humoral immune response to immunization. During Phase 1, cats in Group A (n = 3) received a single intramuscular injection of GonaCon and Group B (n = 3) received a single intramuscular injection of saline. During Phase 2, Group A received a second GonaCon injection and Group B received their initial GonaCon injection. All cats developed GnRH antibodies within 30 days of vaccine administration. The endpoint titre (1:1,024,000) was similar among all cats, and levels remained high throughout the duration of the study. Four cats developed a sterile, painless, self-limiting mass at the site of injection. The mean number of days to mass development was 110.3 (range, 18-249 days). In conclusion, this preliminary study suggests that the EPA-registered GonaCon formulation is safe for continued testing in domestic cats, an accidental revaccination should not increase the risk of a vaccine reaction and the EPA-registered formulation effectively elicits a strong humoral immune response., (© 2016 Blackwell Verlag GmbH.)