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 gestat...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.展开更多
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 a...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.展开更多
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 pre...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.展开更多
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 cli...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.展开更多
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 medicine...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.展开更多
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 ...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.展开更多
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 helpi...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.展开更多
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 advanta...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.展开更多
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）, gr...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.展开更多
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. M...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.展开更多
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 c...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.展开更多
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 a...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.展开更多
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） signal...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.展开更多
The objective of the experiment reported here was to investigate and compare the amino acid(AA)digestibility of distillers' dried grains(DDG), distillers' dried grains with solubles(DDGS), high protein distill...The objective of the experiment reported here was to investigate and compare the amino acid(AA)digestibility of distillers' dried grains(DDG), distillers' dried grains with solubles(DDGS), high protein distillers' dried grains(HP-DDG), and high protein distillers' dried grains with solubles(HP-DDGS) in growing pigs. Five semi-purified diets consisting of DDG, DDGS, HP-DDG, HP-DDGS, and nitrogen-free diet(NFD) were fed to pigs fitted with simple T-cannula for 5 observations per diet. Endogenous losses of AA at the terminal ileum of pigs that received the NFD were used to calculate standardized ileal digestibility(SID) of AA from apparent ileal digestibility(AID) of AA. The AID of Lys in DDGS was lower(P < 0.05) than that in DDG, which was also lower(P < 0.05) than that in HP-DDG. There were no differences in AID of Met among DDG, DDGS and HP-DDGS, but was greater(P < 0.05) in HP-DDG than in DDG or DDGS. The AID of Thr in HP-DDG was greater(P < 0.05) than that in DDGS but not different from that in DDG or HP-DDGS. The branched-chain AA Ile and Leu had greater(P < 0.05) AID in HP-DDG than in DDG, DDGS or HP-DDGS, and there was no difference among DDG, DDGS, and HP-DDGS. The AID of Trp in DDG and DDGS or HP-DDG and HP-DDGS were not different, but the AID of Trp in HP-DDGS was greater(P < 0.05) than that of DDGS. The greatest SID of the indispensable AA was in HP-DDG. Except for Arg and Lys in which DDG had greater(P < 0.05) digestibility, there was no difference between DDG and DDGS in the SID of the indispensable AA. The SID of Lys in DDG was greater(P < 0.05) than that of DDGS but there was no difference between that of DDG and HP-DDGS. Only His, Ile, and Met had lower(P < 0.05) SID in HP-DDGS than HP-DDG within the indispensable AA. The SID of Ala, Asp, Cys, GIu, Gly,Ser and Tyr were lower(P < 0.05) in DDGS than in HP-DDG. There SID of dispensable AA in DDG was not different from that of HP-DDGS. The current study provided apparent and standardized ileal amino acids digestibility values for traditional and high-protein corn distillers' dried grains coproducts for use in formulating swine diets. Amino acid digestibility was generally higher in HP-DDG than in other tested co-products of the dry grind processing of corn for ethanol.展开更多
In this article,one of the grating agencies requested us to incorporate the information,Spanish Government and co-funded by European Union ERDF/ESF,“Investing in your future”,in the acknowledgments section.The corre...In this article,one of the grating agencies requested us to incorporate the information,Spanish Government and co-funded by European Union ERDF/ESF,“Investing in your future”,in the acknowledgments section.The correct acknowledgement is as follows:“This work has been supported by grants of the Spanish Ministry of Health(Fondo de Investigaciones Sanitarias,PI13/00029,Spanish Government and co-funded by European Union ERDF/ESF,“Investing in your future”),Department of Education of Castilla and Leon Regional Government(Grant#LE093U13)and Mutua Madrileña Foundation(Basic research grants 2012)to J.I.R.B.;by Miguel Servet National Program(Ministry of National Health)CP12/03063 and by Gerencia Regional de Salud GRS963/A/2014 to M.L.R.G.We are particularly grateful to Mr.Leonides Alaiz for outstanding animal husbandry.”The authors regret the errors.展开更多
基金financially supported by the National Research Foundation of Korea(NRF-2021R1A5A1033157 for SRC program:382 Comparative medicine Disease Research Center,NRF-2021R1F1A105195313)the Research Institute of Veterinary Science,the BK21 Four for Future Veterinary Medicine Leading Education and Research Center,and a Seoul National University(SNU)grant(#550e2020005)。
文摘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.
文摘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.
文摘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.
基金sponsored by the Deanship of Research at Jordan University of Science and Technology (Grant Number 195/2016)
文摘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.
文摘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.
基金National Research Foundation of Korea(NRF-2017R1A2B3004972)IPET(No.109023–05-5-CG000)The BK21 PLUS Program for Creative Veterinary Science Research
文摘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.
文摘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.
文摘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.
文摘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.
文摘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.
基金the following funds:the National Key R&D Program of China(Project No.2017YFD0502200)the National Natural Science Foundation of China(Project No.31960721)the National Natural Science Foundation of China(Project No.31873034)。
文摘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.
基金The Ohio State University College of Medicine Roessler Research Scholarship(In part)in part by National Cancer Institution of the United States grant No.P30 CA016058(Our histological examination was performed at The OSU Comparative Pathology and Mouse Phenotyping Shared Resource)
文摘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.
文摘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.
文摘The objective of the experiment reported here was to investigate and compare the amino acid(AA)digestibility of distillers' dried grains(DDG), distillers' dried grains with solubles(DDGS), high protein distillers' dried grains(HP-DDG), and high protein distillers' dried grains with solubles(HP-DDGS) in growing pigs. Five semi-purified diets consisting of DDG, DDGS, HP-DDG, HP-DDGS, and nitrogen-free diet(NFD) were fed to pigs fitted with simple T-cannula for 5 observations per diet. Endogenous losses of AA at the terminal ileum of pigs that received the NFD were used to calculate standardized ileal digestibility(SID) of AA from apparent ileal digestibility(AID) of AA. The AID of Lys in DDGS was lower(P < 0.05) than that in DDG, which was also lower(P < 0.05) than that in HP-DDG. There were no differences in AID of Met among DDG, DDGS and HP-DDGS, but was greater(P < 0.05) in HP-DDG than in DDG or DDGS. The AID of Thr in HP-DDG was greater(P < 0.05) than that in DDGS but not different from that in DDG or HP-DDGS. The branched-chain AA Ile and Leu had greater(P < 0.05) AID in HP-DDG than in DDG, DDGS or HP-DDGS, and there was no difference among DDG, DDGS, and HP-DDGS. The AID of Trp in DDG and DDGS or HP-DDG and HP-DDGS were not different, but the AID of Trp in HP-DDGS was greater(P < 0.05) than that of DDGS. The greatest SID of the indispensable AA was in HP-DDG. Except for Arg and Lys in which DDG had greater(P < 0.05) digestibility, there was no difference between DDG and DDGS in the SID of the indispensable AA. The SID of Lys in DDG was greater(P < 0.05) than that of DDGS but there was no difference between that of DDG and HP-DDGS. Only His, Ile, and Met had lower(P < 0.05) SID in HP-DDGS than HP-DDG within the indispensable AA. The SID of Ala, Asp, Cys, GIu, Gly,Ser and Tyr were lower(P < 0.05) in DDGS than in HP-DDG. There SID of dispensable AA in DDG was not different from that of HP-DDGS. The current study provided apparent and standardized ileal amino acids digestibility values for traditional and high-protein corn distillers' dried grains coproducts for use in formulating swine diets. Amino acid digestibility was generally higher in HP-DDG than in other tested co-products of the dry grind processing of corn for ethanol.
文摘In this article,one of the grating agencies requested us to incorporate the information,Spanish Government and co-funded by European Union ERDF/ESF,“Investing in your future”,in the acknowledgments section.The correct acknowledgement is as follows:“This work has been supported by grants of the Spanish Ministry of Health(Fondo de Investigaciones Sanitarias,PI13/00029,Spanish Government and co-funded by European Union ERDF/ESF,“Investing in your future”),Department of Education of Castilla and Leon Regional Government(Grant#LE093U13)and Mutua Madrileña Foundation(Basic research grants 2012)to J.I.R.B.;by Miguel Servet National Program(Ministry of National Health)CP12/03063 and by Gerencia Regional de Salud GRS963/A/2014 to M.L.R.G.We are particularly grateful to Mr.Leonides Alaiz for outstanding animal husbandry.”The authors regret the errors.
