Objective: To evaluate the potential antioxidant and hepatoprotective effects of n-hexane, dichloromethane(DCM), ethyl acetate(EtOAc), n-butanol and aqueous fractions of Moringa oleifera(M. oleifera) leaves methanol e...Objective: To evaluate the potential antioxidant and hepatoprotective effects of n-hexane, dichloromethane(DCM), ethyl acetate(EtOAc), n-butanol and aqueous fractions of Moringa oleifera(M. oleifera) leaves methanol extract against carbon tetrachloride(CCl_4)-induced liver injury in rats. Methods: These fractions were prepared from the M. oleifera leaves methanol extract by solubilization in water and partitioning in n-hexane, EtOAc, DCM and n-butanol. Their phyto-components were identified by GC-MS analysis. The in vitro antioxidant effect of these fractions was carried out by assessment of 1,1-diphenyl-2-picrylhydrazyl scavenging activity. A total of 40 Sprague Dawley rats were allocated into 8 equal groups: group 1 given olive oil(1 m L/kg b.wt.), group 2 injected with CCl_4, group 3 to 7 administered with n-hexane, DCM, EtOAc, n-butanol and aqueous fractions, respectively after CCl_4, group 8 administered with silymarin after CCl_4. The activities of aspartate aminotransferase, alanine aminotransferase, and the levels of total cholesterol, triglycerides, glucose, total proteins and albumin in serum were determined spectrophotometrically. Glutathione reduced, lipid peroxide by-products levels, glutathione-s-transferase and catalase enzyme activities in the liver homogenate were determined by spectrophotometer. Liver specimens were also examined for histopathological alterations under light microscope. Results: The GCMS analysis of different fractions of the M. oleifera leaves methanol extract revealed that n-hexane, DCM, EtOAc, n-butanol, and aqueous fractions contained 17, 22, 23, 19 and 32 compounds, respectively. The percent and the molecular structure of each component in each fraction were identified. The n-butanol and EtOAc fractions exhibited the strongest in vitro antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl. CCl_4 significantly decreased glutathione reduced and total proteins concentration and glutathione-s-transferase and catalase activities but increased lipid peroxide by-products and total cholesterol levels. The n-hexane followed by aqueous and DCM fractions were the most potent to regulate serum enzyme activities and lipid peroxide by-products levels in the liver homogenate. Conclusions: n-hexane, DCM, and aqueous fractions have the highest effectiveness against CCl_4-induced hepatotoxicity. Isolation and purification of the active constituents require further experiments.展开更多
Objective: To analyze the phytochemical constituents, and to explore potential protective effect of the methanol extract of Moringa oleifera(M. oleifera) seeds and Egyptian propolis, each alone or concurrently adminis...Objective: To analyze the phytochemical constituents, and to explore potential protective effect of the methanol extract of Moringa oleifera(M. oleifera) seeds and Egyptian propolis, each alone or concurrently administered on acetic acid-induced ulcerative colitis in rats. Methods: Eight groups of 5 rats each were used: normal control group with distilled water, model group, two groups with M. oleifera seeds(100 and 200 mg/kg), two groups with propolis(50 and 100 mg/kg), one group with concurrent administration of both, and one group with prednisolone(reference drug). Macro-and microscopic picture, ulcer index and lesion scores, oxidative markers, inflammatory mediators, in vitro activity of the inflammatory enzymes and 1, 1-diphenyl-2-picrylhydrazyl free radicals scavenging activity were evaluated. The phytochemical constituents of both extracts were explored by GC-MS analysis. Results: Both treatments modulated the macro-and microscopic picture, decreased the ulcerative index, lesion score, oxidative markers and inflammatory mediators, and inhibited the COX-1 and COX-2 enzymes. Propolis appeared to be powerful free radicals scavenger. A powerful synergistic effect of both treatments in modulating the course of the disease was reported. GCMS analysis of methanol extract of M. oleifera seeds and propolis revealed the presence of 50 and 34 compounds, respectively. Conclusions: M. oleifera seeds and propolis methanol extracts have modulated the course of acetic acid-induced ulcerative colitis. Moreover, both treatments induce a good synergistic effect against the disease. Isolation of the active constituents is recommended.展开更多
Background: In order to improve the efficiency of bovine sperm cryopreservation process, it is important to understand how spermatozoa respond to differences in temperature as well as the ability to recover its own me...Background: In order to improve the efficiency of bovine sperm cryopreservation process, it is important to understand how spermatozoa respond to differences in temperature as well as the ability to recover its own metabolism. The combination between flow cytometry approach and antioxidant enzymes activity allows a more sensible evaluation of sperm cell during cryopreservation. The aim of this study was to evaluate sperm attributes and antioxidant enzymes activity during different stages of cryopreservation process. Semen samples from Holstein bulls(n = 4) were separated in 3 treatments: fresh(37 °C); cooled(5 °C); and thawed. Evaluation occurred at 0 h and2 h after incubation. Membrane integrity, mitochondrial membrane potential(MMP) and DNA damages were evaluated by flow cytometry; activities of antioxidant enzymes such as catalase, superoxide dismutase and gluthatione peroxidase were measured by spectrofotometry.Results: There was an increase in the percentage of sperm with DNA damage in the thawed group, compared to fresh and cooled, and for 2 hs of incubation when compared to 0 h. Considering MMP, there was an increase in the percentage of cells with medium potential in thawed group when compared to fresh and cooled groups. Opposingly,a decrease was observed in the thawed group considering high mitochondrial potential. Also in the thawed group,there was an increase on cells with damaged acrosome and membrane when compared to fresh and cooled groups.Significant correlations were found between antioxidant enzymes activity and membrane or mitochondrial parameters.Conclusion: Based on our results, we conclude that cryopreservation affects cellular and DNA integrity and that the critical moment is when sperm cells are exposed to freezing temperature. Also, our study indicates that intracellular antioxidant machinery(SOD and GPX enzymes) is not enough to control cryodamage.展开更多
Background: In beef cattle, changes in the periovulatory endocrine milieu are associated with fertility and conceptus growth. A large preovulatory follicle(POF) and the resulting elevated concentrations of progesteron...Background: In beef cattle, changes in the periovulatory endocrine milieu are associated with fertility and conceptus growth. A large preovulatory follicle(POF) and the resulting elevated concentrations of progesterone(P4) during diestrus positively affect pregnancy rates. Amino acids(AA) are important components of maternally derived secretions that are crucial for embryonic survival before implantation. The hypothesis is that the size of the POF and the concentration of P4 in early diestrus modulate the endometrial abundance of SLC transcripts related to AA transport and metabolism and subsequently impact luminal concentrations of AA. The follicle growth of Nelore cows was manipulated to produce two experimental groups: large POF and CL(LF-LCL group) and small POF and CL(SF-SCL group). On Day 4(D4; Experiment 1) and Day 7(D7; Experiment 2) after Gn RH-induced ovulation(Gn RH treatment = D0), the animals were slaughtered and uterine tissues and uterine washings were collected. q RT-PCR was used to evaluate the expression levels of AA transporters in D4 and D7 endometrial tissues.The concentrations of AA were quantified in D4 and D7 uterine washings by HPLC.Results: Transcript results show that, on D4, SLC6 A6, SLC7 A4, SLC17 A5, SLC38 A1, SLC38 A7 and SCLY and on D7 SLC1 A4, SLC6 A1, SLC6 A14, SLC7 A4, SLC7 A7, SLC7 A8, SLC17 A5, SLC38 A1, SLC38 A7, SLC43 A2 and DDO were more abundant in the endometria of cows from the LF-LCL group(P < 0.05). In addition, concentrations of AA in the uterine lumen were influenced by the endocrine profiles of the mother. In this context, D4 uterine washings revealed that greater concentrations of taurine, alanine and α-aminobutyric acid were present in SF-SCL(P < 0.05).In contrast, lower concentrations of valine and cystathionine were quantified on D7 uterine washings from SF-SCL cows(P < 0.05).Conclusion: The present study revealed an association between the abundance of transcripts related to AA transport and metabolism in the endometrium and specific periovulatory endocrine profiles related to the receptive status of the mother. Such insights suggest that AAs are involved in uterine function to support embryo development.展开更多
Background: Estradiol(E2) is required for luteolysis in cows and its injection stimulates prostaglandin F2α(PGF2α)release. The main goal of our study was to investigate the ability of endometrial explants and cells ...Background: Estradiol(E2) is required for luteolysis in cows and its injection stimulates prostaglandin F2α(PGF2α)release. The main goal of our study was to investigate the ability of endometrial explants and cells treated with E2 and the calcium ionophore(CI) A23187 to synthesize PGF2α.Results: Treatment with E2 in vivo resulted in a 48.4% increase of PGF2α production by endometrial explants treated in vitro with A23187. Production of PGF2α was better stimulated with A23187 at concentrations of 10-6and10-5mol/L compared with other concentrations used. The concentration of PGF2α for untreated bovine endometrial cell cultures was 33.1 pg/m L, while for cultures treated with E2, A23187, or a combination of E2 and A23187, the PGF2α concentration was 32.5, 92.4 and 145.6 pg/m L, respectively.Conclusions: Treatment with A23187 tended to stimulate PGF2α production. In the presence of E2, A23187 significantly stimulated PGF2α synthesis. It appears that A23187 potentiates the effects of E2 with respect to synthesis of endometrial PGF2α in cattle.展开更多
Objective:To assess the effect of region, season and year of insemination onin-vivo fertility of Italian-Egyptian crossbred buffalo semen.Methods: A total number of 4799 female buffaloes were inseminated by frozen sem...Objective:To assess the effect of region, season and year of insemination onin-vivo fertility of Italian-Egyptian crossbred buffalo semen.Methods: A total number of 4799 female buffaloes were inseminated by frozen semen with at least 50% post-thaw motility of Egyptian-Italian crossbred bulls in three localities in Delta, lower Egypt (El-Behira, El-Sharkia and Damietta) during the period of 2013, 2014 and 2015. The pregnancy rate after two months was evaluated during the four seasons.Results:The rate of pregnancy was significantly (P<0.0001) differ among the three localities. The effect of year of insemination on pregnancy rate was significantly higher during 2014 and 2015 than 2013 in El-Sharkia and El-Behira. But in Damietta, the rate of pregnancy was significantly higher in 2014 than 2013 and 2015. There were no significant differences among seasons in El-Behira and Damietta governorates but there was significant (P<0.05) differences in pregnancy rate in El-Sharkia. It was higher in summer, spring and autumn than in winter.Conclusions:Localities, year of insemination and season of the year have effects on fertility of crossbred Egyptian-Italian buffalo semen.展开更多
Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Me...Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Methods:Two experiments were performed, the first one aimed to evaluate the different concentrations (0, 25, 50, 100 μM) of L-ascorbic acid on embryo developmental rate of buffalo oocytes. The L-ascorbic acid was added to the maturation and culture media. In the second experiment, oocytes were cultured in media with two type of antioxidant (ascorbic acid + cysteamine) or ascorbic acid only.Results:There was a significant increase in cleavage rate at 25, 50 μM than 100 μM and control group. But, the blastocyst rate was higher at 50 μM ascorbic acid than other concentrations (0, 25, 100 μM). Supplementation of ascorbic acid and cysteamine to maturation and cultured media improved embryo development than ascorbic acid alone.Conclusions: Using of 50 μM L-ascorbic acid duringin vitro maturation and development improve the developmental competence of buffalo oocytes, this effect was increase with the presence of cysteamine.展开更多
Objective: To clarify the antioxidant effect of butylated hydroxytoulene (BHT) at different concentrations on cooled and post frozen semen diluted in tris-citrate-fructose egg yolk glycerol and lecithin -based extende...Objective: To clarify the antioxidant effect of butylated hydroxytoulene (BHT) at different concentrations on cooled and post frozen semen diluted in tris-citrate-fructose egg yolk glycerol and lecithin -based extenders. Methods: Forty ejaculates were harvested from four buffalo bulls by means of the artificial vagina. Ejaculated semen samples were diluted with each of the tris citrate-fructose egg yolk glycerol and lecithin-based extender diluents. The semen samples diluted with each of the two extenders were added to pre-warmed dried test tubes containing BHT (prepared in ethanol) to get concentrations at 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mM/mL BHT. These ingredients were put at 37 ℃ for 5 min to allow the proper BHT spermatozoal permeation. The diluted semen samples were cooled to 5 ℃ and then frozen to -196 ℃ in 0.25 mL ministraws before dipping in liquid nitrogen pending its evaluation. Sperm motility, viability, morphology, intact acrosome and membrane integrity were tested. Visual motility was tested using a high power ordinary microscope (at 400 × ) with closed circuit television, and sperm concentration was tested using Neubauer haemocytometer and abnormality % using eosin-nigrosin stain. Spermatozoal membrane integrity was tested using the hypo-osmotic swelling test. The sperm with swollen twisting tail was normally intact. Sperm acrosomal integrity % was tested as mentioned by Watson. Results: Addition of BHT improved (P<0.01) progressive motility, viability, morphology and acrosome as well as plasma membrane integrities at 0.5-2.0 mM/mL depending upon types of used extenders and stages of pre-and post-freezing process. Higher levels of 2.5 and 3.0 mM/mL BHT had a deteriorating (P<0.01) result if compared to the control and all extenders assayed. Conclusions: BHT addition at lower concentration can improve pre-frozen and post-thawed buffalo sperm quality.展开更多
Objective:To investigate the contraceptive effect of intratesticular injection of chlorhexidine gluconate in dogs and compare it with that of zinc gluconate neutralized with arginine.Methods:Twenty-four sexually matur...Objective:To investigate the contraceptive effect of intratesticular injection of chlorhexidine gluconate in dogs and compare it with that of zinc gluconate neutralized with arginine.Methods:Twenty-four sexually mature male mongrel dogs were randomly divided by replicate into four groups(n=6 per group).GroupⅡreceived intra-testicular injection of 2 mL zinc gluconate(10 mg/mL)neutralized with arginine.GroupⅡreceived intratesticular injection of 2 mL chlorhexidine gluconate(5%w/v).GroupⅢreceived intratesticular injection of 2 mL chlorhexidine gluconate(4%w/v).GroupⅣdid not receive any treatment and served as the control group.Testicular morphometry was conducted on day 0,7,15 and 30 after treatment.Semen was collected and evaluated on day 0 and 30.Data were analyzed using repeated measures analysis of variance.Results:There was no difference in the mean values of various parameters between dogs treated with zinc gluconate and those treated with chlorhexidine gluconate at any of the time points.In dogs treated with zinc gluconate or chlorhexidine gluconate,there was a significant increase in the testicular morphometric parameters on day 7 followed by a significant reduction thereafter(day 15 and 30).In contrast,there was no change in any of the parameters in the control untreated dogs during the course of the study.Compared to the pre-treatment values,the mean scrotal circumference and the mean paired testicular volume and testicular weight on day 30 were significantly lower in the treated dogs.Semen samples collected on day 30 from treated dogs were found to be azoospermic,whereas no change in semen quality was observed in the control untreated dogs.Conclusions:Intratesticular injection of chlorhexidine gluconate(5%w/v and 4%w/v)is equally as effective as zinc gluconate neutralized with arginine for chemical contraception in dogs.展开更多
Genetic diversity of two chicken ecotypes from Ismailia-Egypt (ISM) and Taif-Saudi Arabia (TA) was evaluated using 39 microsatellites. DNA was extracted from blood of 25 chickens/ecotype. The number of alleles was 157...Genetic diversity of two chicken ecotypes from Ismailia-Egypt (ISM) and Taif-Saudi Arabia (TA) was evaluated using 39 microsatellites. DNA was extracted from blood of 25 chickens/ecotype. The number of alleles was 157 and 138, the number of alleles/locus averaged 4.2±2.2 and 3.6±1.6, and the highest number of private alleles was 9 and 5 for ISM and TA, respectively. Percentage of shared alleles between the two ecotypes was 45%. This panel of markers is reasonably informative as the mean polymorphic information content for ISM and TA was 0.47±0.21, and 0.41±0.2. Similar average of observed heterozygosity was attained for both ecotypes. Conversely, averages of expected heterozygosity differed between two ecotypes, 0.52±0.23 vs. 0.45±0.21 for ISM and TA. 8 and 12 loci have significantly deviated from HWE of ISM and TA. Estimate of genetic distance was 0.2 and F<sub>ST</sub> index was 0.29. Results showed only 6% of genetic diversity is shared between these two ecotypes.展开更多
Objective:To assess the effects of epidermal growth factor (EGF) on the Egyptian buffalo bull frozen semen, EGF was incorporated at 0 (control), 50, 100, 200 and 400 ng/mL of extender (Bioxcell?).Methods:Semen feature...Objective:To assess the effects of epidermal growth factor (EGF) on the Egyptian buffalo bull frozen semen, EGF was incorporated at 0 (control), 50, 100, 200 and 400 ng/mL of extender (Bioxcell?).Methods:Semen features, spermatozoa biometry, total liberated amounts of enzymes (aspartate transaminase, alanine aminotransferase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) and lipid peroxidation markers (thiobarbituric acid reactive substances, malondialdehyde, glutathione peroxidase, nitric oxide, catalase (CAT) and superoxide dismutase (SOD)) were determined in the spermatozoa-free extracellular extender. Results:Spermatozoa membrane integrity significantly (P<0.05) increased, but DNA integrity decreased with EGF 200 ng/mL. Spermatozoa head (dimensions, area and perimeter), but not shape, as well as acrosome and midpiece measures substantially differed with regard to EGF. Principle piece length and volume markedly decreased (at 100 and 200 ng/mL), while total tail/flagellum length increased (at 50 ng/mL) after EGF supplementation. EGF 50 ng/mL was associated with the decline of nitric oxide levels and catalase enzyme activity, but EGF 100 ng/mL significantly decreased the total liberated amounts of enzymes (aspartate transaminase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) as well as lipid peroxidation markers (thiobarbituric acid reactive substances and malondialdehyde).Conclusion:EGFin vitro supplementation would affect the semen characteristics of buffalo bull with 100 ng/mL counteracted the freezing mediated oxidative stress indicated with the lowest enzymes leakage and lipid peroxidation.展开更多
Objective:To determine the effects of different concentrations of penicillamine, hypotaurine, and epinephrine (PHE) as well as incubation time on motility, hyperactivity and acrosome reaction (AR) of ram spermin vitro...Objective:To determine the effects of different concentrations of penicillamine, hypotaurine, and epinephrine (PHE) as well as incubation time on motility, hyperactivity and acrosome reaction (AR) of ram spermin vitro.Methods: Freshly ejaculated spermatozoa from three ram were collected, pooled and subjected to swim up technique in modified sperm Tyrode's albumin lactate pyruvate medium supplemented with different concentrations of PHE (10, 20, 30, 40, 50, 75 and 100 mM/mL). Then best concentrations were compared and examined for motility, hyperactivity and AR.Results: A high concentrations of PHE (30, 40, 50, 75 and 100mM/mL) showed a significant increase in motility when compared to control immediately after dilution and exist for the first and second hour of incubation period. However, when longer incubation time were used, a significant (P<0.05) decrease in motility was achieved. Similar finding was observed in hyperactivity. Stained semen samples showed a maximum percentage of incomplete AR after 1 h incubation corresponding to 50 and 75 mM/mL of PHE;however, spermatozoa treated with 75 mM/mL had a higher tendency to undergo complete AR after further incubation up to 4 h. A dose dependent relationship was detected where the maximum value of total AR was shown in spermatozoa treated with 75 mM/mL PHE for 4 h. Conclusions: To obtain better motility, hyperactivity and AR, treatment of ram spermatozoa with 75 mM/mL PHE for 4 h before being used in insemination was considered the best concentration of PHE to be used in the process ofin-vitro fertilization.展开更多
Objective:To assess effect of buffalo bull breed on the development and cryotolerence of the in vitro produced embryos.Methods: Three types of frozen semen were adopted;Egyptian, Italian and cross-bred (Egyptian-Itali...Objective:To assess effect of buffalo bull breed on the development and cryotolerence of the in vitro produced embryos.Methods: Three types of frozen semen were adopted;Egyptian, Italian and cross-bred (Egyptian-Italian) breeds were used for in-vitro fertilization and vitrification of their embryos. Oocytes were collected from buffalo ovaries and maturedin vitrofor 24 h, then they were fertilized using the three semen breeds. The produced embryos of morula and blastocysts were vitrified using ethylene glycol and dimethyl sulfoxide then evaluated for their viability after warming.Results: The cleavage and blastocysts rates significantly declined in oocytes fertilized by Egyptian (P<0.01) than in Italian (P<0.05) and crossbred (P<0.05) frozen semen. After embryo vitrification, there were no significant differences among the three breeds in the percentages of morphologically viable embryos evaluated directly after warming and at 24 h post-culture. Conclusions:Thein vitro fertilization response to frozen-thawed semen varies between breeds;however, the resistance of produced embryos to the damage effect of vitrification does not vary.展开更多
Objective:To investigate the effects of non-permeable cryoprotectant,cholesterol-loaded cyclodextrin,when added at different concentrations into cooled and frozen-thawed semen extended with Tris-citrate-fructose egg y...Objective:To investigate the effects of non-permeable cryoprotectant,cholesterol-loaded cyclodextrin,when added at different concentrations into cooled and frozen-thawed semen extended with Tris-citrate-fructose egg yolk glycerol and lecithin-based extenders.Methods:A total of 40 ejaculates from four buffalo bulls were collected using artificial vagina.Ejaculates were extended with one of Tris-citrate-fructose egg yolk glycerol and lecithin-based extenders which contained different concentrations[0(control),0.75,1.50,2.25 and 3.00 mg/mL]of cholesterol-loaded cyclodextrin.The extended semen samples were cooled to 5曟and then frozen slowly to-196曟in 0.25 mL ministraws before being stored in liquid nitrogen pending its evaluation.Sperm motility,live sperm,normal sperm morphology,sperm membrane integrity and acrosome morphology were measured.Results:Supplementation of cholesterol-loaded cyclodextrin improved progressive motility,viability,morphology and acrosome as well as plasma membrane integrities at 1.50-2.25 mg/mL depending upon types of used extenders and stages of pre-and post-freezing process(P<0.01).The best concentration was 1.50 mg/mL at pre-freeze stage and 2.25 mg/mL at post-freezing.However,greater concentration(3.00 mg/mL)of cholesterol-loaded cyclodextrin had a detrimental effect compared to the control group with the two evaluated extenders(P<0.01).Conclusions:Cholesterol-loaded cyclodextrin supplementation at 1.50-2.25 mg/mL concentration could improve pre-frozen and post-thawed buffalo sperm quality.The most suitable concentration is 1.50 mg/mL at pre-freeze stage and 2.25 mg/mL at post-freezing.展开更多
Background: Insufficient production of anti-luteolytic signals by the pre-attachment embryo is considered a major cause of pregnancy failure in cattle.We tested the hypothesis that transfer of multiple blastocysts(n =...Background: Insufficient production of anti-luteolytic signals by the pre-attachment embryo is considered a major cause of pregnancy failure in cattle.We tested the hypothesis that transfer of multiple blastocysts(n = 5/recipient) and progesterone(P4) supplementation amplify anti-luteolytic signaling and reduce embryonic losses in beef cattle.Cows detected in estrus(D0; n = 104) were assigned randomly to receive 150 mg of injectable long-acting P4(iP4) or vehicle(non-iP4) on D4 and transcervical transfer of none or five,grade 1,not-frozen,in vitro-produced blastocysts,on D7.Luteal development and time of structural luteolysis were monitored by ultrasonography.Plasma P4 concentrations were determined on D4,D5 and D7,and daily between D14 and D20.Conceptus signaling was monitored by transcript abundance of interferon-stimulated gene 15(ISG15) in peripheral blood mononuclear cells isolated on D14,D16,D18 and D20.Early embryonic mortality(EEM) was defined as the absence of ISG15 m RNA upregulation over time and/or luteal regression up to D20.Late embryonic mortality(LEM) was defined as the absence of a conceptus with a heartbeat on pregnancy diagnosis at D30(PD30) after observing upregulation of ISG15 mRNA and extension of luteal lifespan.Pregnant cows presented conceptuses with heartbeat at PD30.Results: On D5,iP4-treated cows had P4 concentrations 2.07-fold greater than non-iP4 treated(P < 0.001).On D7,P4 concentrations were similar.Pregnant and LEM animals showed a progressive increase in the abundance of ISG15 from D14 to D20.iP4-treated cows detected pregnant at PD30 had 1.53-fold greater abundance of ISG15 mRNA between D14 and D20 than non-iP4 treated cows(P = 0.05).iP4 doubled the frequency of EEM while it did not affect LEM.At PD30,embryonic survival was 37.0% vs.55.6% for iP4-treated vs.control cows.Majority of pregnant cows(71%)presented only a single viable embryo.Conclusions: A substantial proportion of cows had EEM(31%) and LEM(20%) even after transferring multiple blastocysts.This argues that mortality was due to poor uterine receptivity that could not be reversed by supplemental P4 or overcome by transferring multiple blastocysts.Further,a given uterine environment was not necessarily adequate to all embryos.展开更多
Background: Pregnancy associated glycoproteins form a diverse family of glycoproteins that are variably expressed at different stages of gestation. They are probably involved in immunosuppression of the dam against th...Background: Pregnancy associated glycoproteins form a diverse family of glycoproteins that are variably expressed at different stages of gestation. They are probably involved in immunosuppression of the dam against the feto- maternal placentome. The presence of the products of binucleate cells in maternal circulation has also been correlated with placentogenesis and placental re-modeling. The exact structure and function of the gene product is unknown due to limitations on obtaining purified pregnancy associated glycoprotein preparations. Results: Our study describes an in silico derived 3D model for bubaline pregnancy associated glycoprotein 2. Structure-activity features of the protein were characterized, and functional studies predict bubaline pregnancy associated glycoprotein 2 as an inducible, extra-cellular, non-essential, N-glycosylated, aspartic pro-endopeptidase that is involved in down-regulation of complement pathway and immunity during pregnancy. The protein is also predicted to be involved in nutritional processes, and apoptotic processes underlying fetal morphogenesis and re- modeling of feto-maternal tissues. Conclusion: The structural and functional annotation of buPAG2 shall allow the designing of mutants and inhibitors for dissection of the exact physiological role of the protein.展开更多
Of the tumors diagnosed in the female dogs have the highest mammary neoplasias incidence. These neoplasias can be influenced by environmental contaminants. Despite evidence of pyrethroid toxicity, carcinogenic potenti...Of the tumors diagnosed in the female dogs have the highest mammary neoplasias incidence. These neoplasias can be influenced by environmental contaminants. Despite evidence of pyrethroid toxicity, carcinogenic potential has not yet been sufficiently elucidated, there is a need to investigate their involvement in mammary tumor. In previous studies, pyrethroid residues were detected in female dogs with mammary neoplasia, however was not investigate the influence of this insecticide in the genesis and aggressiveness of mammary cancer. This study aimed to investigate possible relations between pyrethroid residues and aggressiveness of mammary carcinoma in female dogs. Fifty selected female dogs were divided into five groups of 10 animals each: the Control group, female dogs without mammary neoplasia;the groups Luminal A, Luminal B, HER-2 Superexpression and Basal were constituted by female dogs that presented inguinal mammary carcinoma classified immunohistochemically. The aggressiveness of carcinomas was evaluated by immunohistochemistry (HER-2, p63, estrogen receptor). Residual concentrations of the pyrethroids from the mammary gland and fat tissue adjacent to it were determined by HPLC. Data were analyzed by Chi-Square test. Of the all animals, six presented residues of pyrethroids in mammary samples and 10 presented it in fat tissue samples. There was no statistical evidence that pyrethroids are involved in mammary carcinoma aggressiveness in female dogs.展开更多
In Vitro production of swine embryos is a valuable tool to generate clones and genetically modified pigs during a short period of time. However, the efficiency of the existing methods is extremely low and the oocyte q...In Vitro production of swine embryos is a valuable tool to generate clones and genetically modified pigs during a short period of time. However, the efficiency of the existing methods is extremely low and the oocyte quality and quantity represent important obstacles on the success of in Vitro production of embryos. Therefore, the aim of this study was to compare the in Vitro maturation, fertilization and subsequent embryo development rates of oocytes recovered by ovary slicing or follicular aspiration. The oocyte recovery rate (grade 1 COC/ovary) was higher (p = 0.0083) in the slicing group when compared to the aspiration group. No differences were observed between groups regarding in Vitro maturation and early cleavage rates. A higher percentage of oocytes recovered by follicular aspiration reached the blastocyst stage after IVF when compared to the ovary slicing method (p = 0.0395). However, no difference on blastocyst cell number was observed. Although the recovery of oocytes using the slicing technique yielded more grade 1 oocytes per ovary than the aspiration method, the number of oocytes that reached the blastocyst stage after IVF by the slicing method was lower when compared with oocytes obtained by aspiration, as observed by lower blastocyst rates. In conclusion, the follicular aspiration is the method of choice for porcine in Vitro embryo production.展开更多
文摘Objective: To evaluate the potential antioxidant and hepatoprotective effects of n-hexane, dichloromethane(DCM), ethyl acetate(EtOAc), n-butanol and aqueous fractions of Moringa oleifera(M. oleifera) leaves methanol extract against carbon tetrachloride(CCl_4)-induced liver injury in rats. Methods: These fractions were prepared from the M. oleifera leaves methanol extract by solubilization in water and partitioning in n-hexane, EtOAc, DCM and n-butanol. Their phyto-components were identified by GC-MS analysis. The in vitro antioxidant effect of these fractions was carried out by assessment of 1,1-diphenyl-2-picrylhydrazyl scavenging activity. A total of 40 Sprague Dawley rats were allocated into 8 equal groups: group 1 given olive oil(1 m L/kg b.wt.), group 2 injected with CCl_4, group 3 to 7 administered with n-hexane, DCM, EtOAc, n-butanol and aqueous fractions, respectively after CCl_4, group 8 administered with silymarin after CCl_4. The activities of aspartate aminotransferase, alanine aminotransferase, and the levels of total cholesterol, triglycerides, glucose, total proteins and albumin in serum were determined spectrophotometrically. Glutathione reduced, lipid peroxide by-products levels, glutathione-s-transferase and catalase enzyme activities in the liver homogenate were determined by spectrophotometer. Liver specimens were also examined for histopathological alterations under light microscope. Results: The GCMS analysis of different fractions of the M. oleifera leaves methanol extract revealed that n-hexane, DCM, EtOAc, n-butanol, and aqueous fractions contained 17, 22, 23, 19 and 32 compounds, respectively. The percent and the molecular structure of each component in each fraction were identified. The n-butanol and EtOAc fractions exhibited the strongest in vitro antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl. CCl_4 significantly decreased glutathione reduced and total proteins concentration and glutathione-s-transferase and catalase activities but increased lipid peroxide by-products and total cholesterol levels. The n-hexane followed by aqueous and DCM fractions were the most potent to regulate serum enzyme activities and lipid peroxide by-products levels in the liver homogenate. Conclusions: n-hexane, DCM, and aqueous fractions have the highest effectiveness against CCl_4-induced hepatotoxicity. Isolation and purification of the active constituents require further experiments.
文摘Objective: To analyze the phytochemical constituents, and to explore potential protective effect of the methanol extract of Moringa oleifera(M. oleifera) seeds and Egyptian propolis, each alone or concurrently administered on acetic acid-induced ulcerative colitis in rats. Methods: Eight groups of 5 rats each were used: normal control group with distilled water, model group, two groups with M. oleifera seeds(100 and 200 mg/kg), two groups with propolis(50 and 100 mg/kg), one group with concurrent administration of both, and one group with prednisolone(reference drug). Macro-and microscopic picture, ulcer index and lesion scores, oxidative markers, inflammatory mediators, in vitro activity of the inflammatory enzymes and 1, 1-diphenyl-2-picrylhydrazyl free radicals scavenging activity were evaluated. The phytochemical constituents of both extracts were explored by GC-MS analysis. Results: Both treatments modulated the macro-and microscopic picture, decreased the ulcerative index, lesion score, oxidative markers and inflammatory mediators, and inhibited the COX-1 and COX-2 enzymes. Propolis appeared to be powerful free radicals scavenger. A powerful synergistic effect of both treatments in modulating the course of the disease was reported. GCMS analysis of methanol extract of M. oleifera seeds and propolis revealed the presence of 50 and 34 compounds, respectively. Conclusions: M. oleifera seeds and propolis methanol extracts have modulated the course of acetic acid-induced ulcerative colitis. Moreover, both treatments induce a good synergistic effect against the disease. Isolation of the active constituents is recommended.
基金supported by Sao Paulo Research Foundation(FAPESP),process number 2010/18978-9 and 2007/58487-1
文摘Background: In order to improve the efficiency of bovine sperm cryopreservation process, it is important to understand how spermatozoa respond to differences in temperature as well as the ability to recover its own metabolism. The combination between flow cytometry approach and antioxidant enzymes activity allows a more sensible evaluation of sperm cell during cryopreservation. The aim of this study was to evaluate sperm attributes and antioxidant enzymes activity during different stages of cryopreservation process. Semen samples from Holstein bulls(n = 4) were separated in 3 treatments: fresh(37 °C); cooled(5 °C); and thawed. Evaluation occurred at 0 h and2 h after incubation. Membrane integrity, mitochondrial membrane potential(MMP) and DNA damages were evaluated by flow cytometry; activities of antioxidant enzymes such as catalase, superoxide dismutase and gluthatione peroxidase were measured by spectrofotometry.Results: There was an increase in the percentage of sperm with DNA damage in the thawed group, compared to fresh and cooled, and for 2 hs of incubation when compared to 0 h. Considering MMP, there was an increase in the percentage of cells with medium potential in thawed group when compared to fresh and cooled groups. Opposingly,a decrease was observed in the thawed group considering high mitochondrial potential. Also in the thawed group,there was an increase on cells with damaged acrosome and membrane when compared to fresh and cooled groups.Significant correlations were found between antioxidant enzymes activity and membrane or mitochondrial parameters.Conclusion: Based on our results, we conclude that cryopreservation affects cellular and DNA integrity and that the critical moment is when sperm cells are exposed to freezing temperature. Also, our study indicates that intracellular antioxidant machinery(SOD and GPX enzymes) is not enough to control cryodamage.
基金FAPESP 2014/01727-4 to MISSCNPq-481199/2012-8 and FAPESP-2011/03226-4 to MBCNPq 140527/2013-3 to MRF
文摘Background: In beef cattle, changes in the periovulatory endocrine milieu are associated with fertility and conceptus growth. A large preovulatory follicle(POF) and the resulting elevated concentrations of progesterone(P4) during diestrus positively affect pregnancy rates. Amino acids(AA) are important components of maternally derived secretions that are crucial for embryonic survival before implantation. The hypothesis is that the size of the POF and the concentration of P4 in early diestrus modulate the endometrial abundance of SLC transcripts related to AA transport and metabolism and subsequently impact luminal concentrations of AA. The follicle growth of Nelore cows was manipulated to produce two experimental groups: large POF and CL(LF-LCL group) and small POF and CL(SF-SCL group). On Day 4(D4; Experiment 1) and Day 7(D7; Experiment 2) after Gn RH-induced ovulation(Gn RH treatment = D0), the animals were slaughtered and uterine tissues and uterine washings were collected. q RT-PCR was used to evaluate the expression levels of AA transporters in D4 and D7 endometrial tissues.The concentrations of AA were quantified in D4 and D7 uterine washings by HPLC.Results: Transcript results show that, on D4, SLC6 A6, SLC7 A4, SLC17 A5, SLC38 A1, SLC38 A7 and SCLY and on D7 SLC1 A4, SLC6 A1, SLC6 A14, SLC7 A4, SLC7 A7, SLC7 A8, SLC17 A5, SLC38 A1, SLC38 A7, SLC43 A2 and DDO were more abundant in the endometria of cows from the LF-LCL group(P < 0.05). In addition, concentrations of AA in the uterine lumen were influenced by the endocrine profiles of the mother. In this context, D4 uterine washings revealed that greater concentrations of taurine, alanine and α-aminobutyric acid were present in SF-SCL(P < 0.05).In contrast, lower concentrations of valine and cystathionine were quantified on D7 uterine washings from SF-SCL cows(P < 0.05).Conclusion: The present study revealed an association between the abundance of transcripts related to AA transport and metabolism in the endometrium and specific periovulatory endocrine profiles related to the receptive status of the mother. Such insights suggest that AAs are involved in uterine function to support embryo development.
基金funded by Fundao de Amparo à Pesquisa do Estado de So Paulo (FAPESP)
文摘Background: Estradiol(E2) is required for luteolysis in cows and its injection stimulates prostaglandin F2α(PGF2α)release. The main goal of our study was to investigate the ability of endometrial explants and cells treated with E2 and the calcium ionophore(CI) A23187 to synthesize PGF2α.Results: Treatment with E2 in vivo resulted in a 48.4% increase of PGF2α production by endometrial explants treated in vitro with A23187. Production of PGF2α was better stimulated with A23187 at concentrations of 10-6and10-5mol/L compared with other concentrations used. The concentration of PGF2α for untreated bovine endometrial cell cultures was 33.1 pg/m L, while for cultures treated with E2, A23187, or a combination of E2 and A23187, the PGF2α concentration was 32.5, 92.4 and 145.6 pg/m L, respectively.Conclusions: Treatment with A23187 tended to stimulate PGF2α production. In the presence of E2, A23187 significantly stimulated PGF2α synthesis. It appears that A23187 potentiates the effects of E2 with respect to synthesis of endometrial PGF2α in cattle.
文摘Objective:To assess the effect of region, season and year of insemination onin-vivo fertility of Italian-Egyptian crossbred buffalo semen.Methods: A total number of 4799 female buffaloes were inseminated by frozen semen with at least 50% post-thaw motility of Egyptian-Italian crossbred bulls in three localities in Delta, lower Egypt (El-Behira, El-Sharkia and Damietta) during the period of 2013, 2014 and 2015. The pregnancy rate after two months was evaluated during the four seasons.Results:The rate of pregnancy was significantly (P<0.0001) differ among the three localities. The effect of year of insemination on pregnancy rate was significantly higher during 2014 and 2015 than 2013 in El-Sharkia and El-Behira. But in Damietta, the rate of pregnancy was significantly higher in 2014 than 2013 and 2015. There were no significant differences among seasons in El-Behira and Damietta governorates but there was significant (P<0.05) differences in pregnancy rate in El-Sharkia. It was higher in summer, spring and autumn than in winter.Conclusions:Localities, year of insemination and season of the year have effects on fertility of crossbred Egyptian-Italian buffalo semen.
文摘Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Methods:Two experiments were performed, the first one aimed to evaluate the different concentrations (0, 25, 50, 100 μM) of L-ascorbic acid on embryo developmental rate of buffalo oocytes. The L-ascorbic acid was added to the maturation and culture media. In the second experiment, oocytes were cultured in media with two type of antioxidant (ascorbic acid + cysteamine) or ascorbic acid only.Results:There was a significant increase in cleavage rate at 25, 50 μM than 100 μM and control group. But, the blastocyst rate was higher at 50 μM ascorbic acid than other concentrations (0, 25, 100 μM). Supplementation of ascorbic acid and cysteamine to maturation and cultured media improved embryo development than ascorbic acid alone.Conclusions: Using of 50 μM L-ascorbic acid duringin vitro maturation and development improve the developmental competence of buffalo oocytes, this effect was increase with the presence of cysteamine.
文摘Objective: To clarify the antioxidant effect of butylated hydroxytoulene (BHT) at different concentrations on cooled and post frozen semen diluted in tris-citrate-fructose egg yolk glycerol and lecithin -based extenders. Methods: Forty ejaculates were harvested from four buffalo bulls by means of the artificial vagina. Ejaculated semen samples were diluted with each of the tris citrate-fructose egg yolk glycerol and lecithin-based extender diluents. The semen samples diluted with each of the two extenders were added to pre-warmed dried test tubes containing BHT (prepared in ethanol) to get concentrations at 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mM/mL BHT. These ingredients were put at 37 ℃ for 5 min to allow the proper BHT spermatozoal permeation. The diluted semen samples were cooled to 5 ℃ and then frozen to -196 ℃ in 0.25 mL ministraws before dipping in liquid nitrogen pending its evaluation. Sperm motility, viability, morphology, intact acrosome and membrane integrity were tested. Visual motility was tested using a high power ordinary microscope (at 400 × ) with closed circuit television, and sperm concentration was tested using Neubauer haemocytometer and abnormality % using eosin-nigrosin stain. Spermatozoal membrane integrity was tested using the hypo-osmotic swelling test. The sperm with swollen twisting tail was normally intact. Sperm acrosomal integrity % was tested as mentioned by Watson. Results: Addition of BHT improved (P<0.01) progressive motility, viability, morphology and acrosome as well as plasma membrane integrities at 0.5-2.0 mM/mL depending upon types of used extenders and stages of pre-and post-freezing process. Higher levels of 2.5 and 3.0 mM/mL BHT had a deteriorating (P<0.01) result if compared to the control and all extenders assayed. Conclusions: BHT addition at lower concentration can improve pre-frozen and post-thawed buffalo sperm quality.
基金funded by Nanaji Deshmukh Veterinary Science University(grant No.B31/Comp/2018-19 Dated 17/07/2018).
文摘Objective:To investigate the contraceptive effect of intratesticular injection of chlorhexidine gluconate in dogs and compare it with that of zinc gluconate neutralized with arginine.Methods:Twenty-four sexually mature male mongrel dogs were randomly divided by replicate into four groups(n=6 per group).GroupⅡreceived intra-testicular injection of 2 mL zinc gluconate(10 mg/mL)neutralized with arginine.GroupⅡreceived intratesticular injection of 2 mL chlorhexidine gluconate(5%w/v).GroupⅢreceived intratesticular injection of 2 mL chlorhexidine gluconate(4%w/v).GroupⅣdid not receive any treatment and served as the control group.Testicular morphometry was conducted on day 0,7,15 and 30 after treatment.Semen was collected and evaluated on day 0 and 30.Data were analyzed using repeated measures analysis of variance.Results:There was no difference in the mean values of various parameters between dogs treated with zinc gluconate and those treated with chlorhexidine gluconate at any of the time points.In dogs treated with zinc gluconate or chlorhexidine gluconate,there was a significant increase in the testicular morphometric parameters on day 7 followed by a significant reduction thereafter(day 15 and 30).In contrast,there was no change in any of the parameters in the control untreated dogs during the course of the study.Compared to the pre-treatment values,the mean scrotal circumference and the mean paired testicular volume and testicular weight on day 30 were significantly lower in the treated dogs.Semen samples collected on day 30 from treated dogs were found to be azoospermic,whereas no change in semen quality was observed in the control untreated dogs.Conclusions:Intratesticular injection of chlorhexidine gluconate(5%w/v and 4%w/v)is equally as effective as zinc gluconate neutralized with arginine for chemical contraception in dogs.
文摘Genetic diversity of two chicken ecotypes from Ismailia-Egypt (ISM) and Taif-Saudi Arabia (TA) was evaluated using 39 microsatellites. DNA was extracted from blood of 25 chickens/ecotype. The number of alleles was 157 and 138, the number of alleles/locus averaged 4.2±2.2 and 3.6±1.6, and the highest number of private alleles was 9 and 5 for ISM and TA, respectively. Percentage of shared alleles between the two ecotypes was 45%. This panel of markers is reasonably informative as the mean polymorphic information content for ISM and TA was 0.47±0.21, and 0.41±0.2. Similar average of observed heterozygosity was attained for both ecotypes. Conversely, averages of expected heterozygosity differed between two ecotypes, 0.52±0.23 vs. 0.45±0.21 for ISM and TA. 8 and 12 loci have significantly deviated from HWE of ISM and TA. Estimate of genetic distance was 0.2 and F<sub>ST</sub> index was 0.29. Results showed only 6% of genetic diversity is shared between these two ecotypes.
文摘Objective:To assess the effects of epidermal growth factor (EGF) on the Egyptian buffalo bull frozen semen, EGF was incorporated at 0 (control), 50, 100, 200 and 400 ng/mL of extender (Bioxcell?).Methods:Semen features, spermatozoa biometry, total liberated amounts of enzymes (aspartate transaminase, alanine aminotransferase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) and lipid peroxidation markers (thiobarbituric acid reactive substances, malondialdehyde, glutathione peroxidase, nitric oxide, catalase (CAT) and superoxide dismutase (SOD)) were determined in the spermatozoa-free extracellular extender. Results:Spermatozoa membrane integrity significantly (P<0.05) increased, but DNA integrity decreased with EGF 200 ng/mL. Spermatozoa head (dimensions, area and perimeter), but not shape, as well as acrosome and midpiece measures substantially differed with regard to EGF. Principle piece length and volume markedly decreased (at 100 and 200 ng/mL), while total tail/flagellum length increased (at 50 ng/mL) after EGF supplementation. EGF 50 ng/mL was associated with the decline of nitric oxide levels and catalase enzyme activity, but EGF 100 ng/mL significantly decreased the total liberated amounts of enzymes (aspartate transaminase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) as well as lipid peroxidation markers (thiobarbituric acid reactive substances and malondialdehyde).Conclusion:EGFin vitro supplementation would affect the semen characteristics of buffalo bull with 100 ng/mL counteracted the freezing mediated oxidative stress indicated with the lowest enzymes leakage and lipid peroxidation.
文摘Objective:To determine the effects of different concentrations of penicillamine, hypotaurine, and epinephrine (PHE) as well as incubation time on motility, hyperactivity and acrosome reaction (AR) of ram spermin vitro.Methods: Freshly ejaculated spermatozoa from three ram were collected, pooled and subjected to swim up technique in modified sperm Tyrode's albumin lactate pyruvate medium supplemented with different concentrations of PHE (10, 20, 30, 40, 50, 75 and 100 mM/mL). Then best concentrations were compared and examined for motility, hyperactivity and AR.Results: A high concentrations of PHE (30, 40, 50, 75 and 100mM/mL) showed a significant increase in motility when compared to control immediately after dilution and exist for the first and second hour of incubation period. However, when longer incubation time were used, a significant (P<0.05) decrease in motility was achieved. Similar finding was observed in hyperactivity. Stained semen samples showed a maximum percentage of incomplete AR after 1 h incubation corresponding to 50 and 75 mM/mL of PHE;however, spermatozoa treated with 75 mM/mL had a higher tendency to undergo complete AR after further incubation up to 4 h. A dose dependent relationship was detected where the maximum value of total AR was shown in spermatozoa treated with 75 mM/mL PHE for 4 h. Conclusions: To obtain better motility, hyperactivity and AR, treatment of ram spermatozoa with 75 mM/mL PHE for 4 h before being used in insemination was considered the best concentration of PHE to be used in the process ofin-vitro fertilization.
文摘Objective:To assess effect of buffalo bull breed on the development and cryotolerence of the in vitro produced embryos.Methods: Three types of frozen semen were adopted;Egyptian, Italian and cross-bred (Egyptian-Italian) breeds were used for in-vitro fertilization and vitrification of their embryos. Oocytes were collected from buffalo ovaries and maturedin vitrofor 24 h, then they were fertilized using the three semen breeds. The produced embryos of morula and blastocysts were vitrified using ethylene glycol and dimethyl sulfoxide then evaluated for their viability after warming.Results: The cleavage and blastocysts rates significantly declined in oocytes fertilized by Egyptian (P<0.01) than in Italian (P<0.05) and crossbred (P<0.05) frozen semen. After embryo vitrification, there were no significant differences among the three breeds in the percentages of morphologically viable embryos evaluated directly after warming and at 24 h post-culture. Conclusions:Thein vitro fertilization response to frozen-thawed semen varies between breeds;however, the resistance of produced embryos to the damage effect of vitrification does not vary.
文摘Objective:To investigate the effects of non-permeable cryoprotectant,cholesterol-loaded cyclodextrin,when added at different concentrations into cooled and frozen-thawed semen extended with Tris-citrate-fructose egg yolk glycerol and lecithin-based extenders.Methods:A total of 40 ejaculates from four buffalo bulls were collected using artificial vagina.Ejaculates were extended with one of Tris-citrate-fructose egg yolk glycerol and lecithin-based extenders which contained different concentrations[0(control),0.75,1.50,2.25 and 3.00 mg/mL]of cholesterol-loaded cyclodextrin.The extended semen samples were cooled to 5曟and then frozen slowly to-196曟in 0.25 mL ministraws before being stored in liquid nitrogen pending its evaluation.Sperm motility,live sperm,normal sperm morphology,sperm membrane integrity and acrosome morphology were measured.Results:Supplementation of cholesterol-loaded cyclodextrin improved progressive motility,viability,morphology and acrosome as well as plasma membrane integrities at 1.50-2.25 mg/mL depending upon types of used extenders and stages of pre-and post-freezing process(P<0.01).The best concentration was 1.50 mg/mL at pre-freeze stage and 2.25 mg/mL at post-freezing.However,greater concentration(3.00 mg/mL)of cholesterol-loaded cyclodextrin had a detrimental effect compared to the control group with the two evaluated extenders(P<0.01).Conclusions:Cholesterol-loaded cyclodextrin supplementation at 1.50-2.25 mg/mL concentration could improve pre-frozen and post-thawed buffalo sperm quality.The most suitable concentration is 1.50 mg/mL at pre-freeze stage and 2.25 mg/mL at post-freezing.
基金FAPESP-2015/26215–9 and CAPES-33002010047P6 to TM.FAPESP-2011/03226–4 to MBfunding bodies had no participation on the study,collection,analysis,interpretation of data nor in writing the manuscript
文摘Background: Insufficient production of anti-luteolytic signals by the pre-attachment embryo is considered a major cause of pregnancy failure in cattle.We tested the hypothesis that transfer of multiple blastocysts(n = 5/recipient) and progesterone(P4) supplementation amplify anti-luteolytic signaling and reduce embryonic losses in beef cattle.Cows detected in estrus(D0; n = 104) were assigned randomly to receive 150 mg of injectable long-acting P4(iP4) or vehicle(non-iP4) on D4 and transcervical transfer of none or five,grade 1,not-frozen,in vitro-produced blastocysts,on D7.Luteal development and time of structural luteolysis were monitored by ultrasonography.Plasma P4 concentrations were determined on D4,D5 and D7,and daily between D14 and D20.Conceptus signaling was monitored by transcript abundance of interferon-stimulated gene 15(ISG15) in peripheral blood mononuclear cells isolated on D14,D16,D18 and D20.Early embryonic mortality(EEM) was defined as the absence of ISG15 m RNA upregulation over time and/or luteal regression up to D20.Late embryonic mortality(LEM) was defined as the absence of a conceptus with a heartbeat on pregnancy diagnosis at D30(PD30) after observing upregulation of ISG15 mRNA and extension of luteal lifespan.Pregnant cows presented conceptuses with heartbeat at PD30.Results: On D5,iP4-treated cows had P4 concentrations 2.07-fold greater than non-iP4 treated(P < 0.001).On D7,P4 concentrations were similar.Pregnant and LEM animals showed a progressive increase in the abundance of ISG15 from D14 to D20.iP4-treated cows detected pregnant at PD30 had 1.53-fold greater abundance of ISG15 mRNA between D14 and D20 than non-iP4 treated cows(P = 0.05).iP4 doubled the frequency of EEM while it did not affect LEM.At PD30,embryonic survival was 37.0% vs.55.6% for iP4-treated vs.control cows.Majority of pregnant cows(71%)presented only a single viable embryo.Conclusions: A substantial proportion of cows had EEM(31%) and LEM(20%) even after transferring multiple blastocysts.This argues that mortality was due to poor uterine receptivity that could not be reversed by supplemental P4 or overcome by transferring multiple blastocysts.Further,a given uterine environment was not necessarily adequate to all embryos.
文摘Background: Pregnancy associated glycoproteins form a diverse family of glycoproteins that are variably expressed at different stages of gestation. They are probably involved in immunosuppression of the dam against the feto- maternal placentome. The presence of the products of binucleate cells in maternal circulation has also been correlated with placentogenesis and placental re-modeling. The exact structure and function of the gene product is unknown due to limitations on obtaining purified pregnancy associated glycoprotein preparations. Results: Our study describes an in silico derived 3D model for bubaline pregnancy associated glycoprotein 2. Structure-activity features of the protein were characterized, and functional studies predict bubaline pregnancy associated glycoprotein 2 as an inducible, extra-cellular, non-essential, N-glycosylated, aspartic pro-endopeptidase that is involved in down-regulation of complement pathway and immunity during pregnancy. The protein is also predicted to be involved in nutritional processes, and apoptotic processes underlying fetal morphogenesis and re- modeling of feto-maternal tissues. Conclusion: The structural and functional annotation of buPAG2 shall allow the designing of mutants and inhibitors for dissection of the exact physiological role of the protein.
文摘Of the tumors diagnosed in the female dogs have the highest mammary neoplasias incidence. These neoplasias can be influenced by environmental contaminants. Despite evidence of pyrethroid toxicity, carcinogenic potential has not yet been sufficiently elucidated, there is a need to investigate their involvement in mammary tumor. In previous studies, pyrethroid residues were detected in female dogs with mammary neoplasia, however was not investigate the influence of this insecticide in the genesis and aggressiveness of mammary cancer. This study aimed to investigate possible relations between pyrethroid residues and aggressiveness of mammary carcinoma in female dogs. Fifty selected female dogs were divided into five groups of 10 animals each: the Control group, female dogs without mammary neoplasia;the groups Luminal A, Luminal B, HER-2 Superexpression and Basal were constituted by female dogs that presented inguinal mammary carcinoma classified immunohistochemically. The aggressiveness of carcinomas was evaluated by immunohistochemistry (HER-2, p63, estrogen receptor). Residual concentrations of the pyrethroids from the mammary gland and fat tissue adjacent to it were determined by HPLC. Data were analyzed by Chi-Square test. Of the all animals, six presented residues of pyrethroids in mammary samples and 10 presented it in fat tissue samples. There was no statistical evidence that pyrethroids are involved in mammary carcinoma aggressiveness in female dogs.
基金supported financially by Fundacao de Amparo a Pesquisa do Estado de Sao Paulo(FAPESP).
文摘In Vitro production of swine embryos is a valuable tool to generate clones and genetically modified pigs during a short period of time. However, the efficiency of the existing methods is extremely low and the oocyte quality and quantity represent important obstacles on the success of in Vitro production of embryos. Therefore, the aim of this study was to compare the in Vitro maturation, fertilization and subsequent embryo development rates of oocytes recovered by ovary slicing or follicular aspiration. The oocyte recovery rate (grade 1 COC/ovary) was higher (p = 0.0083) in the slicing group when compared to the aspiration group. No differences were observed between groups regarding in Vitro maturation and early cleavage rates. A higher percentage of oocytes recovered by follicular aspiration reached the blastocyst stage after IVF when compared to the ovary slicing method (p = 0.0395). However, no difference on blastocyst cell number was observed. Although the recovery of oocytes using the slicing technique yielded more grade 1 oocytes per ovary than the aspiration method, the number of oocytes that reached the blastocyst stage after IVF by the slicing method was lower when compared with oocytes obtained by aspiration, as observed by lower blastocyst rates. In conclusion, the follicular aspiration is the method of choice for porcine in Vitro embryo production.
