Molecular characteristics,antibiogram and prevalence of multi-drug resistant Staphylococcus aureus (MDRSA) isolated from milk obtained from culled dairy cows and from cows with acute clinical mastitis

To study the molecular characteristics, antibiogram and prevalence of multi-drug resistant Staphylococcus aureus (S. aureus) (MDRSA) isolated from milk obtained from culled dairy cows and from cows with acute clinical mastitis.MethodsBacteria were cultured from 188 quarter milk samples obtained from cows before culling (n = 139) and from cows affected with acute mastitis (n = 49) belonging to 10 dairy farms. The bacteria were identified using colony morphology, Gram staining and biochemical characteristics. S. aureus isolates were then subjected to molecular characterization using PCR targeting 16S rRNA and mecA gene to identify Methicillin resistant S. aureus (MRSA). The antibiogram of all isolates was performed using the Kirby-Bauer disk diffusion method against 10 commonly used antibiotics in dairy farms.ResultsS. aureus was isolated from 19 (13.7%) samples obtained from culled cows and 11 (22.4%) samples obtained from cows with acute mastitis. In both culled cows and cows with acute mastitis, in vitro antibiogram revealed that 100% of S. aureus isolates were resistant to erythromycin, penicillin G, streptomycin, doxycyclin, and trimethoprim/sulpha. The prevalence of MRSA in milk of culled cows and cows with acute mastitis was 26.3% and 18.2%, respectively, with an overall prevalence of 3.7% among all samples. All MRSA isolates were completely resistant to all tested antibiotics. All MRSA isolates were positive for the presence of the mecA gene.ConclusionsMRSA carrying the mecA gene were isolated from mastitic milk from dairy cows in Jordan for the first time. MRSA may pose a potential health risk to the public, farm workers and veterinarians.

Clinical Effects of a Plant Extract Mixture Containing Rhus verniciflua and Other Herbs in Tumor Bearing Dogs

Dog owners are increasingly seeking treatment when their pets develop cancers. As in human cancer patients, dogs with cancer are commonly treated with complementary and alternative therapies, including herbal medicines and nutritional supplements. A novel antitumor agent was developed from six different herbs including Rhus verniciflua (Rv-PEM01). The components were established from traditional herbal medicine and designed to affect antitumor activity and maintain host immune function. Previous studies identified anti-proliferative activity in human, murine and canine cancer cell lines. In this clinical study the safety and tolerability of Rv-PEM01 were evaluated in pet dogs with spontaneously occurring cancers. Twelve dogs were treated orally daily for 30 days in escalating dose (4 - 10 mg/kg orally once daily) cohorts. Rv-PEM01 was well tolerated;only transient mild elevations in BUN were observed in 2 dogs. Although tumor response was not a primary endpoint for this study, stable disease was maintained for 30 days in 5 (42%) of the dogs. In conclusion, Rv-PEM01 was found to be safe and well tolerated in the dosage range tested. Future studies should evaluate higher dosages of Rv-PEM01 in dogs with cancer, and specifically address other potential benefits of Rv-PEM01 in canine cancer patients, including correlative assessments of immune function, quality of life and owner satisfaction.

Additively manufactured Ti–Ta–Cu alloys for the next-generation load-bearing implants

Bacterial colonization of orthopedic implants is one of the leading causes of failure and clinical complexities for load-bearing metallic implants. Topical or systemic administration of antibiotics may not offer the most efficient defense against colonization, especially in the case of secondary infection, leading to surgical removal of implants and in some cases even limbs. In this study, laser powder bed fusion was implemented to fabricate Ti3Al2V alloy by a 1:1 weight mixture of CpTi and Ti6Al4V powders. Ti-Tantalum(Ta)–Copper(Cu) alloys were further analyzed by the addition of Ta and Cu into the Ti3Al2V custom alloy. The biological,mechanical, and tribo-biocorrosion properties of Ti3Al2V alloy were evaluated. A 10 wt.% Ta(10Ta) and 3 wt.% Cu(3Cu) were added to the Ti3Al2V alloy to enhance biocompatibility and impart inherent bacterial resistance. Additively manufactured implants were investigated for resistance against Pseudomonas aeruginosa and Staphylococcus aureus strains of bacteria for up to 48 h. A 3 wt.% Cu addition to Ti3Al2V displayed improved antibacterial efficacy, i.e.78%–86% with respect to CpTi. Mechanical properties for Ti3Al2V–10Ta–3Cu alloy were evaluated, demonstrating excellent fatigue resistance, exceptional shear strength, and improved tribological and tribo-biocorrosion characteristics when compared to Ti6Al4V. In vivo studies using a rat distal femur model revealed improved early-stage osseointegration for alloys with10 wt.% Ta addition compared to CpTi and Ti6Al4V. The 3 wt.% Cu-added compositions displayed biocompatibility and no adverse infammatory response in vivo. Our results establish the Ti3Al2V–10Ta–3Cu alloy’s synergistic effect on improving both in vivo biocompatibility and microbial resistance for the next generation of load-bearing metallic implants.

Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein(RNP)electroporation

Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising cost.Furthermore,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in cattle.In this point of view,electroporation in embryos has been risen as an alternative.Results First,editing efficiency of our electroporation methods were tested for embryos.Presence of mutation on embryo was confirmed by T7E1 assay.With first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,respectively.In case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for BLG.Afterwards,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing.Thirteen recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double KO.Ten surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both genes.Conclusions These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied.Finally,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.

Robust Machine Learning Technique to Classify COVID-19 Using Fusion of Texture and Vesselness of X-Ray Images

Manual investigation of chest radiography(CXR)images by physicians is crucial for effective decision-making in COVID-19 diagnosis.However,the high demand during the pandemic necessitates auxiliary help through image analysis and machine learning techniques.This study presents a multi-threshold-based segmentation technique to probe high pixel intensity regions in CXR images of various pathologies,including normal cases.Texture information is extracted using gray co-occurrence matrix(GLCM)-based features,while vessel-like features are obtained using Frangi,Sato,and Meijering filters.Machine learning models employing Decision Tree(DT)and RandomForest(RF)approaches are designed to categorize CXR images into common lung infections,lung opacity(LO),COVID-19,and viral pneumonia(VP).The results demonstrate that the fusion of texture and vesselbased features provides an effective ML model for aiding diagnosis.The ML model validation using performance measures,including an accuracy of approximately 91.8%with an RF-based classifier,supports the usefulness of the feature set and classifier model in categorizing the four different pathologies.Furthermore,the study investigates the importance of the devised features in identifying the underlying pathology and incorporates histogrambased analysis.This analysis reveals varying natural pixel distributions in CXR images belonging to the normal,COVID-19,LO,and VP groups,motivating the incorporation of additional features such as mean,standard deviation,skewness,and percentile based on the filtered images.Notably,the study achieves a considerable improvement in categorizing COVID-19 from LO,with a true positive rate of 97%,further substantiating the effectiveness of the methodology implemented.

Development of genome engineering technologies in cattle: from random to specific

The production of transgenic farm animals(e.g., cattle) via genome engineering for the gain or loss of gene functions is an important undertaking. In the initial stages of genome engineering, DNA micro-injection into one-cell stage embryos(zygotes) followed by embryo transfer into a recipient was performed because of the ease of the procedure.However, as this approach resulted in severe mosaicism and has a low efficiency, it is not typically employed in the cattle as priority, unlike in mice. To overcome the above issue with micro-injection in cattle, somatic cell nuclear transfer(SCNT) was introduced and successfully used to produce cloned livestock. The application of SCNT for the production of transgenic livestock represents a significant advancement, but its development speed is relatively slow because of abnormal reprogramming and low gene targeting efficiency. Recent genome editing technologies(e.g.,ZFN, TALEN, and CRISPR-Cas9) have been rapidly adapted for applications in cattle and great results have been achieved in several fields such as disease models and bioreactors. In the future, genome engineering technologies wil accelerate our understanding of genetic traits in bovine and wil be readily adapted for bio-medical applications in cattle.

Lactobacillus rhamnosus GR-1 attenuates foodborne Bacillus cereus-induced NLRP3 inflammasome activity in bovine mammary epithelial cells by protecting intercellular tight junctions

Background:Bacillus cereus is an important pathogen that causes human food poisoning,specifically diarrhea and vomiting.B.cereus can also induce mastitis in dairy cows and has a strong survival ability in milk,as it cannot be inactivated by high-temperature short-time pasteurization.Therefore,B.cereus can enter the market through pasteurized milk and other dairy products,imposing enormous hidden dangers on food safety and human health.Results:In this study,B.cereus 2101(BC)was isolated from milk samples of cows with mastitis.BC grew rapidly with strong hemolysis,making it difficult to prevent mastitis and ensure food security.MAC-T cells were treated with BC and/or Lactobacillus rhamnosus GR-1(LGR-1).Pretreatment with LGR-1 protected the integrity of tight junctions and the expression of zonula occludens-1(ZO-1)and occludin destroyed by BC.Furthermore,LGR-1 pretreatment reduced the expression of NOD-like receptor family member pyrin domain-containing protein 3(NLRP3),caspase recruitment and activation domain(ASC),Caspase-1 p20,gasdermin D(GSDMD)p30,inflammatory factors(interleukin(IL)-1βand IL-18),and cell death induced by BC.Moreover,LGR-1 pretreatment reduced NLRP3 inflammasome activity and increased expressions of ZO-1 and occludin induced by lipopolysaccharides(LPS)+ATP stimulation.MAC-T cells were transfected with NLRP3 si RNA or MCC950 and/or treated with BC and/or LGR-1.NLRP3-si RNA transfection and MCC950 attenuated BC-induced NLRP3 inflammasome activity.Expression of inflammatory cytokines and cell death suggested that the inflammatory pathway might play an important role in the induction of the NLRP3 inflammasome by BC and the protection of LGR-1.Conclusions:These results suggest that LGR-1 might be a probiotic alternative to antibiotics and could be administered to prevent mastitis in dairy cows,thus ensuring food security.

Intra-articular transplantation of porcine adipose-derived stem cells for the treatment of canine osteoarthritis: A pilot study

AIM: To test whether intra-articular injection of porcine adipose-derived stem cells(ADSCs) can treat canine osteoarthritis(OA).METHODS: To enroll in this study dogs must have stifle joint OA that had lasted ≥ 3 mo and been treated with OA medication without significant improvement. Three dogs fulfilled these criteria and were thus subjects for ADSCs treatment. ADSCs were isolated from abdominal adipose tissue of a 2-mo-old female Yorkshire pig. Their stem cell marker expression was examined by immunofluorescence staining. For treatment, 5 million ADSCs were injected into the diseased joint of each dog. In the next 48 h, the patient was observed for signs of inflammatory and allergic reactions. Thepatient was then discharged to the owner and, at 2, 6, and 12 wk, followed up with orthopedic assessment, owner questionnaire, X-ray imaging, and force-plate gait analysis.RESULTS: Porcine ADSCs expressed mesenchymal stem cell markers CD90 and CD105. Injection of porcine ADSCs into canine stifle joints did not cause any inflammatory or allergic reactions. Orthopedic evaluation found improvements in two dogs, particularly at the longest time point. Owners' evaluation found increased capacity and decreased pain in all three dogs' activities such as walking and running. Radiographic evaluation did not find statistically significant differences before and after treatment. Force-plate analysis found significant improvements in all three dogs after treatment.CONCLUSION: Xenotransplantation of ADSCs for the treatment of OA is feasible. Further studies are needed to validate this novel treatment modality, which can then be implemented for the routine treatment of OA in veterinary medicine.

Fetal vs adult mesenchymal stem cells achieve greater gene expression, but less osteoinduction

AIM: To investigate adenoviral transduction in mesenchymal stem cells(MSCs) and effects on stemness in vitro and function as a cell therapy in vivo.METHODS: Bone marrow-derived adult and fetal MSC were isolated from an equine source and expanded in monolayer tissue culture. Polyethylenimine(PEI)-mediated transfection of pc DNA3-e GFP or adenoviral transduction of green fluorescent protein(GFP) was evaluated in fetal MSCs. Adenoviral-mediated transduction was chosen for subsequent experiments. All experiments were carried out at least in triplicate unless otherwise noted. Outcome assessment was obtained by flow cytometry or immunohystochemistry and included transduction efficiency, cell viability, stemness(i.e., cell proliferation, osteogenic and chondrogenic cell differentiation), and quantification of GFP expression. Fetal and adult MSCs were then transduced with an adenoviral vector containing the gene for the bone morphogenic protein 2(BMP2). In vitro BMP2 expression was assessed by enzyme linked immunosorbent assay. In addition, MSC-mediated gene delivery of BMP2 was evaluated in vivo in an osteoinduction nude mouse quadriceps model. New bone formation was evaluated by microradiography and histology.RESULTS: PEI provided greater transfection and viability in fetal MSCs than other commercial chemical reagents. Adenoviral transduction efficiency was superior to PEI-mediated transfection of GFP in fetal MSCs(81.3% ± 1.3% vs 35.0% ± 1.6%, P < 0.05) and was similar in adult MSCs(78.1% ± 1.9%). Adenoviral transduction provided significantly greater expression of GFP in fetal than adult MSCs(7.4 ± 0.1 vs 4.4 ± 0.3 millions of mean fluorescence intensity units, P < 0.01) as well as significantly greater in vitro BMP2 expression(0.16 pg/cell-day vs 0.10 pg/cell-day, P < 0.01). Fraction of fetal MSC GFP positive cells decreased significantly faster than adult MSCs(1.15% ± 0.05% vs 11.4% ± 2.1% GFP positive at 2 wk post-transduction, P < 0.05). Cell proliferation and osteogenic differentiation in vitrowere not affected by Ad transduction in both fetal and adult MSCs, but fetal MSCs had reduced chondrogenic differentiation in vitro when compared to adult(P < 0.01). Chondrogenic differentiation was also significantly reduced in Ad-GFP transduced cells(P < 0.05). AdBMP2 transduced adult MSCs induced new bone formation in more thighs than Ad-BMP2 transduced fetal MSCs(83% vs 17% of the six treated thighs per group, P < 0.05) and resulted in increased femur midshaft diameter due to greater extent of periosteal new bone(1.57 ± 0.35 mm vs 1.27 ± 0.08 mm, P < 0.05).CONCLUSION: Fetal MSCs may be genetically manipulated ex vivo with adenoviral vectors. Nonetheless, the abbreviated expression of the exogenous gene may limit their applications in vivo.

Zinc oxide nanoparticles accelerate the healing of methicillin-resistant Staphylococcus aureus(MRSA)-infected wounds in rabbits

Objective:To synthesize zinc oxide nanoparticles(ZnONPs)and evaluate their antibacterial and wound healing effects against wounds infected with methicillin-resistant Staphylococcus aureus(MRSA).Methods:ZnONPs were prepared by sol-gel method and characterized by X-ray diffraction(XRD)analysis and scanning electron microscopy(SEM).A total of 18 rabbits were divided into three groups:the ZnONPs group,the gentamicin group and the control group.A wound of 3 cm^(2) was inflicted on each rabbit and contaminated with MRSA inoculum.Treatment was started from the fourth day post-surgery.Wound healing,microbiological analysis,and histopathological analysis were performed to assess the efficacy of ZnONPs ointment.Results:XRD analysis confirmed the hexagonal wurtzite structure of the ZnONPs with an average crystallite size of 29.23 nm.SEM revealed discoid-shaped ZnONPs with a rough surface and an average size of 48.36 nm.Energy-dispersive X-ray analysis confirmed the purity of ZnONPs.Moreover,the particle size ranged from 100-700 nm with a high agglomeration trend.Treatment with ZnONPs promoted MRSA-infected wound healing.In addition,ZnONPs showed a good antibacterial effect as evidenced by a dose-dependent increase in the zone of inhibition.Conclusions:ZnONPs accelerate the healing of MRSA-infected wounds.Therefore,it can be explored for the treatment of MRSA infection.

Evaluation of the Aurora Kinase Inhibitor, ZM447439, in Canine Malignant Lymphoid Cells <i>in Vitro</i>

Aurora kinases play an important role in the cell cycle. These enzymes help establish mitotic spindles by directing centrosome duplication and separation and by regulating the spindle assembly checkpoint thereby helping control cytokinesis. An over-expression of aurora kinases has been reported in a variety of human tumors. In this study, we identified the expression of aurora-A and aurora-B kinases in canine malignant lymphoid cells. We also evaluated the effects of the aurora kinase inhibitor (ZM447439), and found that this inhibitor decreases cell viability, increases DNA content change, and leads to apoptosis in canine B- and T-cell lymphoid cell lines. The lymphotoxicity induced by ZM447439 in these canine lymphoid cell lines suggests that further in vivo evaluation of aurora kinase inhibitors as a potential treatment for canine malignant lymphoid tumors is warranted.

Initial joint bleed volume in a delayed on-demand treatment setup correlates with subsequent synovial changes in hemophilic mice

Background: Hemophilic arthropathy is a debilitating morbidity of hemophilia caused by recurrent joint bleeds. We investigated if the joint bleed volume, before initiation of treatment, was linked to the subsequent degree of histopathological changes and the development of bone pathology in a mouse model of hemophilic arthropathy.Methods: FVIII knock-out(F8-KO) mice were dosed with a micro-CT blood pool agent prior to induction of hemarthrosis. Eight hours after induction, the bleed volume was quantified with micro computed tomography(micro-CT) and recombinant FVIII treatment initiated. On Day 8, inflammation in the knees was characterized by fluorescence molecular tomography. On Day 14, knee pathology was characterized by micro-CT and histopathology. In a second study, contrast agent was injected into the knee of wild-type(WT) mice, followed by histopathological evaluation on Day 14.Results: The average joint bleed volume before treatment was 3.9 mm3. The inflammation-related fluorescent intensities in the injured knees were significantly increased on Day 8. The injured knees had significantly increased synovitis scores, vessel counts, and areas of hemosiderin compared to un-injured knees. However, no cartilage-or bone pathology was observed. The bleed volume before initiation of treatment correlated with the degree of synovitis and was associated with high fluorescent intensity on Day 8. In F8-KO and WT mice, persistence of contrast agent in the joint elicited morphological changes.Conclusion: When applying a delayed on-demand treatment regimen to hemophilic mice subjected to an induced knee hemarthrosis, the degree of histopathological changes on Day 14 reflected the bleed volume prior to initiation of treatment.

The Influence of Culture Medium Type on Cellular Phenotype of Canine Adipose Derived Stem Cells

Canine adipose derived stem cells (ASCs) hold a great promise for the therapy of osteoarthritis in veterinary medicine. Current therapy is an autologous, stromal vascular fraction. Allogeneic ASCs provide many advantages, including efficient, cost-effective treatments while eliminating a surgical procedure in a diseased animal. Cultured ASCs can be expanded and characterized, allowing selection of desirable qualities. Use of allogeneic ASCs requires selection of a culture medium that provides consistent, desirable cellular products. The supplements within a medium can greatly influence cellular phenotypes. We hypothesized that medium type influenced cellular phenotype, allowing selection of a specified cellular product for clinical applications. We evaluated ASCs derived from adipose tissue of six dogs, assessing mRNA expression of proinflammatory: interleukin-1b, cyclooxygenase-2, and anti-inflammatory mediators: tissue inhibitor metalloproteinase-2 and interleukin -1 receptor antagonist, via quantitative RT-PCR prior to, and following culture in five cell culture media: basic cell growth medium (BGM), Keratinocyte N acetyl-L-cysteine supplemented (KNAC) medium, Multipotent Adult Progenitor Cell (MAPC) medium, serum free medium (SFM) and xeno-free medium. Major histocompatability complex I (MHCI), major histocompatability complex II (MHCII), CD44 and CD90 immunophenotypes were assessed via flow cytometry analysis. Tri-lineage differentiation (bone, adipose and cartilage tissue) was utilized to verify multipotency. SFM and xeno-free culture conditions did not produce cell expansion sufficient to assess phenotype. ASCs prior to culture had wide variability in all mediator levels, while culturing in the remaining conditions resulted in more predictable expression levels of inflammatory mediators, with a decrease in all levels. Cultured ASCs retained expression of cell surface markers MHCI, CD44 and CD90, while decreasing MHCII expression levels. KNAC and MAPC medium conditions consistently produced tri-lineage differentation;BGM, SFM and xeno-free medium did not. Culture condition will influence phenotype of ASCs, and should be selected according to the intended therapeutic effect.

IKKβ overexpression together with a lack of tumour suppressor genes causes ameloblastic odontomas in mice

Odontogenic tumours are a heterogeneous group of lesions that develop in the oral cavity region and are characterized by the formation of tumoural structures that differentiate as teeth. Due to the diversity of their histopathological characteristics and clinical behaviour, the classification of these tumours is still under debate. Alterations in morphogenesis pathways such as the Hedgehog,MAPK and WNT/β-catenin pathways are implicated in the formation of odontogenic lesions, but the molecular bases of many of these lesions are still unknown. In this study, we used genetically modified mice to study the role of IKKβ(a fundamental regulator of NF-κB activity and many other proteins) in oral epithelial cells and odontogenic tissues. Transgenic mice overexpressing IKKβ in oral epithelial cells show a significant increase in immune cells in both the oral epithelia and oral submucosa. They also show changes in the expression of several proteins and mi RNAs that are important for cancer development. Interestingly, we found that overactivity of IKKβ in oral epithelia and odontogenic tissues, in conjunction with the loss of tumour suppressor proteins(p53, or p16 and p19), leads to the appearance of odontogenic tumours that can be classified as ameloblastic odontomas, sometimes accompanied by foci of secondary ameloblastic carcinomas. These tumours show NF-κB activation and increased β-catenin activity.These findings may help to elucidate the molecular determinants of odontogenic tumourigenesis and the role of IKKβ in the homoeostasis and tumoural transformation of oral and odontogenic epithelia.

IN VIVO TRANS-RECTAL ULTRASOUND-COUPLED NEAR-INFRARED OPTICAL TOMOGRAPHY OF INTACT NORMAL CANINE PROSTATE

This is the first tomography-presentation of the optical properties of a normal canine prostate,in vivo,in its native intact environment in the pelvic canal.The imaging was performed by trans-rectal near-infrared(NIR)optical tomography in steady-state measurement at 840 nm on three sagittal planes across the right lobe,middle-line,and left lobe,respectively,of the prostate gland.The NIR imaging planes were position-correlated with concurrently applied trans-rectal ultrasound,albeit there was no spatial prior employed in the NIR tomography reconstruction.The reconstructed peak absorption coefficients of the prostate on the three planes were 0.014,0.012,and 0.014mm^(−1).The peak reduced scattering coefficients were 5.28,5.56,and 6.53 mm^(−1).The peak effective attenuation coefficients were 0.45,0.43,and 0.50 mm^(−1).The absorption and effective attenuation coefficients were within the ranges predictable at 840 nm by literature values which clustered sparsely from 355 nm to 1064 nm,none of which were performed on a canine prostate with similar conditions.The effective attenuation coefficients of the gland were shown to be generally higher in the internal aspects than in the peripheral aspects,which is consistent with the previous findings that the urethral regions were statistically more attenuating than the capsular regions.

Reversal of maternal obesity attenuates hypoxia and improves placental development in the preeclamptic-like BPH/5 mouse model

Background:Women with obesity have higher risk of adverse pregnancy outcomes,including preeclampsia(PE).Late-gestational hypertension,aberrant fetoplacental development,and fetal growth restriction(FGR),hallmarks of PE,are observed spontaneously in BPH/5 mice.Similar to obese preeclamptic women,BPH/5 mice have higher visceral white adipose tissue(WAT)and circulating leptin.We hypothesized that attenuation of maternal obesity and serum leptin in pregnant BPH/5 mice will improve fetoplacental development by decreasing hypoxia markers and leptin expression at the maternal-fetal interface.Methods:To test this hypothesis,BPH/5 mice were fed ad libitum(lib)and pair-fed(PF)to C57 ad lib controls beginning at embryonic day(e)0.5.Hypoxia-related genes,hypoxia inducible factor(Hif)1α,stem cell factor(Scf),heme oxygenase-1(Ho-1),leptin(Lep),and leptin receptor(LepR)were assessed in e7.5 implantation sites.Results:BPH/5 ad lib had 1.5 to 2-fold increase in Hif1α,Scf,and Ho-1 mRNA and a greater than 3-fold increase in leptin mRNA vs.C57 that was attenuated with PF.Exogenous leptin promoted Hif1αand Ho-1 mRNA expression in e7.5 decidua in vitro.While hypoxic conditions in vitro did not change decidual leptin mRNA.Furthermore,BPH/5 PF mice demonstrated improved fetal and placental outcomes later in gestation,with greater placental vascular area by e18.5 and attenuation of FGR.Conclusion:In conclusion,pair-feeding BPH/5 mice beginning at conception may improve placental vasculature formation via decreased leptin and hypoxia-associated markers in this model.Future investigations are needed to better determine the effect of hypoxia and leptin on pregnancy outcomes in obese pregnant women.

In vivo percutaneous reflectance spectroscopy of fatty liver development in rats suggests that the elevation of the scattering power is an early indicator of hepatic steatosis

This study assessed whether there was a scat tering spectral mar ker quantifiable by reflectance measurements that could indicate early development of hepatic steatosis in rats for potential applications to pre procurement organ evaluation.Sixteen rats were fed a methionine choline-deficient(MCD)diet and eight rats were fed a normal diet.Direct assessment of the liver parenchyma of rats in vivo was performed by percut aneous reflect ance spectroscopy using a single fiber probe at the beginning of diet-intake and arbitrary post-diet-intake times up to 11 weeks to render longitudinal comparison.Histological sampling of the liver over the duration of diet adm inistration was performed on two MCD diet treated rats and one control rat eutha-nized after reflectance spectroscopy measurement.The images of hematoxylin/eosin-stained liver specimens were analyzed morphometrically to evahuate the lipid size changes associated with the level of steatosis.The MCD-diet-treated group(n=16)had mild steatosis in seven rats,moderate in three rats,severe in six rats,and no other significant pathology.No control rats(n=8)developed hepatic steatosis.Among the parameters retrieved from per-SfS,only the scat tering power(can be either positive or negative)appeared to be statist ically diferent between MCD-treated and control livers.The scattering power for the 16 MCD-diet-treated livers at the time of euthanasia and presenting various levels of steatosis was 033±0.21,in comparison to 0.036±0.25 of the eight control livers(p=0.0189).When evaluated at days 12 and 13 combined,the scattering power of the 16 MCD-diet-treated livers was 032±0.17,in comparison to 0.10±0.11 of the eight control livers(p=0.0017).All of four MCD-treated livers harvested at days 12 and 13 presented mild steatosis with sub-micron size lipid droplets,even though none of the MCD-treated livers were sonogr aphically remarkable for fatty changes.The elevation of the scattering power may be a valuable marker indicating early hepatic steatosis before the steatosis is sonographically detectable.

Evaluation of Immunomodulatory Activity of the Herbals Formula Viranur, Turmeric （Curcuma heyneana Val.） and Phyllanthus （Phyllanthus niruri L.） in Layer Chicken Vaccinated with Avian Influenza

The aim of this research was to know the effect of herbal as immunomodulator on chicken layer vaccinated with avian influenza. A total of 60 chickens were alloted into three treatment groups： control group （KA）, group KB and group KC, with 20 chicken each group. All the chickens were vaccinated with Newcastle disease （ND） in the age of one week, and a week later they were vaccinated with avian influenza （AI）. The chickens in group KB were drunken with herbal solution containing of 5 g turmeric （Curcuma heyneana Val.） and 25 g phyllanthus （Phyllanthus niruri L.） and group KC were drunken with herbal solution containing of 36 g the herbals formula Viranur and 25 g phyllanthus （Phyllanthus niruri L.）, respectively for four weeks. Thirty days after AI vaccinated, all of chicken were weighed and necropsied. Samples from bursa of Fabricius, tymus and spleen were taken for weighing and histopathological examination. The weight indexs of bursa of Fabricius, tymus and spleen were not significantly different between control group and treatment group in the considered statistically significance （P 〉 0.05）, but the treatment groups （KB and KC） had higher weight index. The histopathologically changes of spleen in both control group and treatment groups were not different, although in the group KC, in bursa of Fabricius, there was lymphocyte increase in its lymphoid follicles; and in the group KB and KC, the tymus were more widening in the cortex than medulla. The conclusion of this study showed that the herbals can stimulate lymphocyte activity.

Pilot Study Evaluating the Use of a Commercially Available Oral Nutritional Supplement in the Management of Chronic Kidney Disease in Cats

Purpose: To determine if cats with chronic kidney disease (CKD) would willingly consume an oral nutritional supplement formula (NS-CKD) and to assess associated effects on select clinical and biochemical parameters. Methods: Client-owned cats with CKD classified as International Renal Interest Society (IRIS) stage 2 (n = 7), IRIS stage 3 (n = 12), or IRIS stage 4 (1 cat) were classified by the owners as having normal or variable appetites. The cats were offered 30 ml NS-CKD for 14 days concurrently with a meal in a separate bowl and the amount of the NS-CKD consumed daily was recorded. Bodyweight, physical examination, and serum biochemical profiles were assessed on Days 0, 7, and 14. Results: Greater than 50% of the NS-CKD was consumed by 14 of 20 (70%) cats and 12 of 20 cats (60%) consumed >80% of the NS-CKD. The total volume of NS-CKD consumed over the course of the study was statistically greater for the cats classified by owners as having normal appetite (P = 0.046). Increases in body weight were noted for 9 of 14 cats (64.3%) that ingested >50% of the NS-CKD and 1 of 6 cats (16.7%), that ingested ≤ 50% (p = 0.1409) and the group mean % change in body weight was greater in the cats that ingested >50% of the NS-CKD (P = 0.023). The volume of NS-CKD consumed correlated to the % change increases in serum bicarbonate concentration (R = 0.4998;P = 0.02) and was weakly correlated to % change decreases in serum phosphorus concentration (R = 0.0406;P = 0.08). Conclusions: In this pilot study, the NS-CKD was accepted by most cats, no adverse effects were noted, and several findings suggest that the product was associated with ameliorating some metabolic complications which suggest it could be considered in the management of cats with CKD.

T follicular helper expansion and humoral-mediated rejection are independent of the HVEM/BTLA pathway

The molecular pathways contributing to humoral-mediated allograft rejection are poorly defined. In this study, we assessed the role of the herpesvirus entry mediator/B- and T-lymphocyte attenuator （HVEM/BTLA） signalling pathway in the context of antibody-mediated allograft rejection. An experimental setting was designed to elucidate whether the blockade of HVEM/BTLA interactions could modulate de novo induction of host antidonor-specific antibodies during the course of graft rejection. To test this hypothesis, fully allogeneic major histocompatibility complex-mismatched skin grafts were transplanted onto the right flank of recipient mice that were treated with isotype control, anti-CD40L or modulatory antibodies of the HVEM/BTLA signalling pathway. The frequencies of CD4 T follicular helper （Tfh） cells （B220-, CD4＋ CXCR5＋ PD-lhigh）, extrafollicular helper cells （B220-, CD4＋ CXCR5- PD-1＋ and PD-1-） and germinal centre （GC） B cells （B220＋Fas＋ GL7＋） were analysed by flow cytometry in draining and non-draining lymph nodes at day 10 post transplantation during the acute phase of graft rejection. The host antidonor isotype-specific humoral immune response was also assessed. Whereas blockade of the CD40/CD40L pathway was highly effective in preventing the allogeneic humoral immune response, antibody-mediated blockade of the HVEM/BTLA-interacting pathway affected neither the expansion of Tfh cells nor the expansion of GC B cells. Consequently, the course of the host antidonor antibody-mediated response proceeded normally, without detectable evidence of impaired development. In summary, these data indicate that HVEM/BTLA interactions are dispensable for the formation of de novo host antidonor isotype-specific antibodies in transplantation.