Effect of Intra-ovarian Injection of Mesenchymal Stem Cells or its Conditioned Media on Repeated OPU-IVEP Outcomes in Jersey Heifers and Its Relationship with Follicular Fluid Inflammatory Markers.

Background: Repeated Ovum Pick Up (OPU) could have a detrimental effect on ovarian function, reducing In Vitro Embryo Production (IVEP). The present study examined the therapeutic effect of adipose-derived Mesenchymal Stem Cells (MSCs) or its Conditioned Medium (ConM) on ovarian trauma following repeated OPU. Resolvin E1 (RvE1) and Interleukin-12 (IL-12) were investigated as biomarkers. Methods: Jersey heifers (n=8) experienced 11 OPU sessions including 5 pre-treatment and 6 treatment sessions. Heifers received intra-ovarian administration of MSCs or ConM (right ovary) and Dulbecco's Modified Phosphate Buffer Saline (DMPBS; left ovary) after OPU in sessions 5 and 8 and 2 weeks after session 11. The concentrations of RvE1 and IL-12 in follicular fluid was evaluated on sessions 1, 5, 6, 9, and 4 weeks after session 11. Following each OPU session, the IVEP parameters were recorded. Results: Intra-ovarian administration of MSCs, ConM, and DMPBS did not affect IVEP parameters (p>0.05). The concentration of IL-12 in follicular fluid increased at the last session of pre-treatment (Session 5; p<0.05) and remained elevated throughout the treatment period. There was no correlation between IL-12 and IVEP parameters (p>0.05). However, RvE1 remained relatively high during the pre-treatment and decreased toward the end of treatment period (p<0.05). This in turn was associated with decline in some IVEP parameters (p<0.05). Conclusion: Intra-ovarian administration of MSCs or ConM during repeated OPU did not enhance IVEP outcomes in Bos taurus heifers. The positive association between RvE1 and some of IVEP parameters could nominate RvE1 as a promising biomarker to predict IVEP parameters following repeated OPU.

Synchronization of estrus using progesterone injections followed by human menopausal gonadotropin in ewes.

Serial progesterone injections followed by human menopausal gonadotropin (hMG), instead of equine chorionic gonadotropin (eCG), were used to synchronize estrus in ewes. Shal ewes (n = 189) were assigned into five groups and each group was divided into two sub-groups to receive gonadotropins including eCG (300 IU; intra-muscular) or hMG (one ampoule; subcutaneously, SC). All ewes received prostaglandin (PG) F2α six days after introducing ram (day 0). Ewes received 0 (control), one, two, three or four injections of progesterone (50.00 mg; SC), 72 hr apart. The first progesterone was injected at the time of PG injection. Ewes in treatment groups received gonadotropins 48 hr after the last progesterone injection. Control group ewes received gonadotropins, at the time of PG injection. Mating was recorded after introducing fertile rams. Data were analyzed using GLM and GENMOD procedures in SAS. The incidence of estrus was less in control and ewes received a single progesterone (34.20%) compared to ewes received two (64.10%), three (81.10%) and four injections (68.40%) of progesterone. Time to estrus was earlier in control (45.70 ± 4.41 hr) than progesterone-treated groups (63.60 ± 1.79 hr). Fertility (51.30%) and fecundity (78.40%) of ewes received three progesterone injections were significantly greater than other progesterone-treated groups. There was no significant difference in reproductive indices between eCG and hMG sub-groups. In conclusion, during the non-breeding season, three injections of progesterone, three days apart, starting six days after ram exposure, in association with hMG, 48 hr after the last progesterone injection, could provide a sound reproductive performance in Shal ewes.

Metabolic enhancement of the one carbon metabolism (OCM) in bovine oocytes IVM increases the blastocyst rate: evidences for a OCM checkpoint.

The one carbon metabolism (OCM) has a primary role in the process of oocyte maturation. In this study bovine oocytes were cultured for 24 h, up to MII stage, with standard medium supplemented or not with 8 metabolic enhancers of the OCM and the MII and blastocyst rate were compared. Additional analyses were performed on matured oocytes, cumulus cells, zygotes and blastocysts. The OCM supplementation increased the blastocyst rate derived from in vitro fertilization. The mitochondrial mass and DNMT3a protein expression were increased whereas DNA fragmentation decreased in matured oocytes. DNA methylation in female pronucleus of zygotes was increased. The supplementation did not directly affect the redox balance as ROS and GSH in matured oocytes and homocysteine in the spent medium were unchanged. The supplementation of the oocytes with metabolic enhancers of the OCM may increase the yield from the culture, likely due to improved DNA methylation and epigenetic programming. The lack of effects on MII rate with huge differences appearing at the blastocyst stage suggest the existence of a OCM metabolic check point that hampers oocytes progression to blastocyst post-fertilization, if they were not properly primed at the time of maturation.

Superovulation of high-producing Holstein lactating dairy cows with human recombinant FSH and hMG.

Superovulation of high-producing dairy cows is a challenging subject in dairy farms with respect to the cost, dose and type of gonadotropin. The objectives of this study were to compare three gonadotropin products: Folltropin-V® (highly purified FSH with porcine origin), Cinnal-f® (recombinant human FSH) and Menotropins® (hMG) for superovulation in high-producing Holstein lactating dairy cows and to investigate the pregnancy outcomes achieved following transferring embryos recovered from donors treated with different gonadotropins. Healthy high-producing Holstein lactating dairy cows (n = 30; milk production: 46.35 ± 8.78 kg; parity: 2-4; days in milk: 80-130 days) without any puerperal problems were selected as donors. On Day 10 after estrus (Day 0 of superovulation), donors (10 cows in each experimental groups) received Folltropin-V® (400 mg NIH, dissolved in 20 ml), Cinnal-f® (20 vials; each vial of 1 ml contains 75 IU Follitropin alfa) and Menotropins ® (20 ampules; each ampule of 1 ml contains 75 IU FSH and 75 IU LH), administered twice daily, in decreasing doses (4,4; 3,3; 2,2; 1,1 ml), over 4 days. On Day 2 of superovulation, donors received 3 doses of prostaglandin F2α analogue, 6 h apart. They were inseminated twice with a frozen semen at 12 and 24 h after standing estrus. Concurrent with the second insemination, donors received 2500 IU hCG (Karma Pharmatech GmbH, Germany). On Day 7 after standing estrus, superovulatory responses (number of CLs, total ova/embryos and transferable embryos) were recorded and Code 1 embryos, recovered from each treated donors, were transferred to synchronized heifers. Pregnancy was detected on Day 30 and 60 after AI. Gestation length, the number and weight of live births were recorded. Data were analyzed using Proc GLM, Proc Mixed and Proc Genmod of SAS. The respective number of corpora lutea, total number of ova/embryos and transferable embryos were not different among donors received Cinnal-f (25.5 ± 3.01, 11.2 ± 2.77, 5.1 ± 0.86), Menotropins (24.0 ± 3.21, 9.0 ± 2.04, 6.3 ± 1.74) and Folltropin-V (20.3 ± 3.21, 8.9 ± 1.90, 5.1 ± 1.16; P > 0.05). Pregnancy rates on Day 30 was similar among treatment groups (P > 0.05). However, pregnancy rates on Day 60 and the number of calves born healthy was less in heifers that received embryos from Cinnal-f treated donors (P < 0.05). In conclusion, Cinnal-f and Menotropins could provide similar superovulatory response to Folltropin-V for superovulation of high-producing Holstein lactating dairy cows.

Epigenotoxic Effect of Dimethyl Sulfoxide on Buffalo Somatic Cells and Buffalo-Bovine Interspecies Somatic Cell Nuclear Transfer Embryos.

OBJECTIVE: In the present study, we investigated the possible epigenotoxic effect of dimethyl sulfoxide (DMSO) on buffalo fibroblast cells and on reconstructed oocytes during buffalo-bovine interspecies somatic cell nuclear transfer (iSCNT) procedure and its effect on rate and quality of blastocyst which derived from these reconstructed oocytes. MATERIALS AND METHODS: In this experimental study, cell viability of buffalo fibroblasts was assessed after exposure to various concentration (0.5, 1, 2 and 4%) of DMSO using MTS assay. The epigenetic effect of DMSO was also assessed in terms of DNA methylation in treated cells by flowcytometry. Reconstructed oocytes of buffalo-bovine iSCNT exposed for 16 hours after activation to non-toxic concentration of DMSO (0.5%) to investigate the respective level of 5-methylcytosine, cleavage and blastocyst rates and gene expression (pluripotent genes: OCT4, NANOG, SOX2, and trophectodermal genes: CDX2 and TEAD4) of produced blastocysts. RESULTS: Supplementation of culture medium with 4% DMSO had substantial adverse effect on the cell viability after 24 hours. DMSO, at 2% concentration, affected cell viability after 48 hours and increased DNA methylation and mRNA expression of DNMT3A in fibroblast cells. Exposure of reconstructed oocytes to 0.5% DMSO for 16 hours post activation did not have significant effect on DNA methylation, nor on the developmental competency of reconstructed oocyte, however, it decreased the mRNA expression of NANOG in iSCNT blastocysts. CONCLUSION: Depending on the dose, DMSO might have epigenotoxic effect on buffalo fibroblast cells and reconstructed oocytes and perturb the mRNA expression of NANOG in iSCNT blastocysts.

Effect of DNA and Histone Methyl Transferase Inhibitors on Outcomes of Buffalo-Bovine Interspecies Somatic Cell Nuclear Transfer.

Somatic cell nuclear transfer (SCNT) derived embryos suffer from abnormal epigenetic reprogramming, which handicaps pre- and postimplantation development. It was hypothesized that epigenetic modifiers, including zebularine (DNA methyltransferase inhibitors) and BIX-01294 (histone methyltransferase inhibitors), could decrease the respective levels of 5-methylcytosine and H3K9me2 in reconstructed oocytes (RO). Accordingly, we investigated whether treating RO with zebularine and BIX-01294 for 16 hours after activation could improve developmental competence and quality of buffalo-bovine interspecies SCNT (iSCNT) embryos. Treatment of RO with zebularine but not BIX-01294 significantly increased two-cell formation at 16 hours postactivation. Conversely, early cleaved embryos had significantly lower rate of blastocyst formation in zebularine treated RO compared to their counterparts in control and BIX-01294 groups. Treatment of RO with zebularine and BIX-01294 did not improve blastocyst rate of buffalo-bovine iSCNT embryos compared to their control counterparts. However, these two epigenetic drugs might have some beneficial effects on buffalo-bovine iSCNT compared to bovine SCNT embryos. The quality of iSCNT blastocysts was improved due to significant expression of OCT4 and CDX2 in BIX-01294 and CDX2 in zebularine treated RO. Furthermore, treatment of RO with zebularine and BIX-01294 did not affect DNA fragmentation in derived blastocysts against control group. In conclusion, treatment with zebularine and BIX-01294 did not enhance developmental competence of iSCNT embryos, but may have some beneficial effects on epigenetic makeup and quality of derived blastocysts.

Superovulatory response following FSH, eCG-FSH and hMG and pregnancy rates following transfer of hatched blastocyst embryos with different diameter and shape in dromedary camel.

The objectives of this study were to compare three superovulation protocols and to investigate the effect of embryo diameter and shape on pregnancy rates in dromedary camel. In Experiment 1, following follicular wave synchronization of donors using two injections of GnRH (25 µg, im), superovulation was initiated 2 days after the second GnRH (Day 0 of experiment). Then, donors were assigned into three groups (n = 5) and received 1) FSH alone (390 mg), 2) eCG (1000 IU; single dose on Day 0) and FSH (330 mg) and 3) hMG (17.5 ampules). FSH and hMG were administered twice daily, in decreasing doses, for 5.5 days. Thirty-six hours after the last dose of FSH or hMG, donors were mated once. On Day 4 of superovulation, the diameter and number of follicles and on Day 7.5 after mating, total number of corpora lutea and follicles ≥9 mm were recorded using ultrasound examination. On Day 8.5 after mating, embryos were recovered non-surgically. In Experiment 2, on Day 6.5 after inducing ovulation, recipients (n = 139) received a single 7.5 days old hatched blastocyst. The diameter and shape (round, oval, collapsed) of transferred embryos were recorded. Pregnancy was detected on Day 25 after embryo transfer. Data were analyzed using Proc GLM, Proc FREG, Proc CORR and Proc Genmod of SAS. The number of corpora lutea was greater in FSH (22.4 ± 2.25) than eCG-FSH (11.6 ± 2.58) and hMG (7 ± 3.19; P < 0.01). Total number of embryos was higher in FSH (16.2 ± 2.72) than hMG (1.6 ± 1.17; P < 0.01). In FSH group, there was correlation between the ratio of follicles >6 mm/follicles ≤6 mm on Day 4 of superovulation and total number of corpora lutea (r = 0.9; P < 0.05) and embryos (r = 0.91; P < 0.05). In the latter group, there was also correlation between corpora lutea and number of embryos (r = 0.92; P < 0.05). Transfer of embryos collected from different superovulation protocols did not affect pregnancy rates (P > 0.05). The diameter of hatched blastocysts had significant effect on pregnancy rates (P < 0.05). Medium size embryos (601-985 µm) produced greater pregnancy rates (50.8%) compared to small (275-600 µm; 25%) and large (986-1571 µm; 32%) embryos (P < 0.05). In conclusion, FSH protocol provided better and predictable superovulatory response. Greater pregnancy rates could be achieved by transferring medium size hatched blastocysts in dromedary camel.

Supplementation of tris-based extender with plasma egg yolk of six avian species and camel skim milk for chilled preservation of dromedary camel semen.

The present study was conducted to investigate a suitable source (Expt. 1) and concentration (Expt. 2) of plasma egg yolk (PEY) and concentration of camel skim milk (CSM; Expt. 3) to supplement tris based extender for chilled storage of dromedary camel semen. In Expt. 1, PEY (20%) of six avian species (domestic chicken, domestic duck, Japanese quail, partridge, pigeon and guinea fowl) was added to semen extender. In Expt. 2, different concentrations (0, 10, 20, 30 and 40%) of selected PEY from Expt.1 were added to semen extender. In both Expt. 1 and 2, CSM remained constant (20%). In Expt. 3, semen extender was supplemented with different concentrations of CSM (0, 20, 40, 60 and 80%) while the concentration of PEY remained constant. The sperm viability parameters were assessed at 6, 12 and 24h following chilled storage. In Expt. 1, progressive forward motility (PFM) of diluted semen supplemented with pigeon PEY was similar to domestic duck and Japanese quail PEYs (P>0.05) and superior to other PEYs (P<0.05). In Expt. 2, PFM following the addition of 20% pigeon PEY was similar to 10 and 30% (P>0.05) and greater than 0 and 40% (P<0.05). In Expt. 3, total motility, PFM and live percentage of sperm were better in 20% compared to 40, 60 and 80% CSM (P<0.05). In the last experiment, PFM in 20% was better than 0% CSM (P<0.05). In conclusion, pigeon PEY at the concentration of 20% and CSM at the concentration of 20% could provide beneficial effect on some of the sperm viability parameters during chilled storage of dromedary camel semen.

Comparison of virulence factors and biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections.

The aim of this study was to find different prevalence of genes involved in the biofilm formation process and to assess the phenotypic and genotypic markers of biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections. In this study, 215 S. aureus strains were collected from human and dairy cow's infections. The biofilm forming capacity of the strains was evaluated using a colorimetric microtiter plate assay. The genes encoding microbial surface components, recognizing adhesive matrix molecules (MSCRAMMs) (ebpS, eno, fib, fnbA, fnbB, cna and bap), and the intracellular adhesion (ica) genes (icaA, and icaD) were targeted by polymerase chain reaction (PCR)-based method. Approximately 70% of the isolates produced biofilm. Among these, 59.3% were producers of weakly adherent biofilms while 34.8% and 5.8% produced moderate and strong biofilms, respectively. The most prevalent gene was icaD found in 88.4% of the isolates, followed by icaA, fib and eno found in 87.9%, 75.8% and 75.3% of the isolates, respectively. The bap gene was not detected in any of the isolates. The prevalence of ebpS and fnbA genes among bovine isolates were significantly higher than those in human isolates, whilst the prevalence of cna gene was significantly higher in the human isolates. In this study, a high prevalence of biofilm production was found among S. aureus strains isolated from human and bovine infections. Most biofilm producing isolates were positive for MSCRAMM, icaA, and icaD genes.

Effect of camel milk on blood sugar and lipid profile of patients with type 2 diabetes: a pilot clinical trial.

BACKGROUND: It has been shown that camel milk consumption has a definite decreasing effect on the prevalence of diabetes. However, most of these studies were conducted on patients with type 1 diabetes, whereas studies on patients with type 2 diabetes mellitus (T2DM) are limited. In vitro experiments have shown that camel milk was able to decrease blood glucose concentration. OBJECTIVES: The purpose of this study was to investigate effects of camel and cow milk on blood sugar, lipid profile, and blood pressure of patients with T2DM. PATIENTS AND METHODS: In a randomized single-blinded controlled clinical trial, 20 patients with T2DM were randomly allocated into two groups. Participants consumed 500 mL of either camel milk (intervention group) or cow milk (control group) daily for two months. RESULTS: Mean of insulin concentration was significantly increased from 64.59 to 84.03 pmol/L in the camel milk group during the study (P < 0.05). No significant differences were shown in fasting blood sugar, lipid profile, and blood pressure between the two groups at the end of study. There was significant increase in homeostasis model assessment of insulin resistance (HOMA-IR) during the study in both groups, but no significant difference was seen between the two groups. CONCLUSIONS: Camel milk increased insulin level in patients with T2DM and might contribute to glycemic control in T2DM.

Comparison between two portal laparoscopy and open surgery for ovariectomy in dogs.

Ovariectomy (OVE) is a routine surgical procedure for neutering in small animal practice. Laparoscopy is a new surgical technique which contains advantages such as less trauma, smaller incision and excellent visualization than traditional open surgery. The present study was conducted to examine the feasibility and safety of laparoscopic procedure through two portal comparing with the conventional open surgery for OVE in healthy female bitches (n=16). Dogs were divided in two equal groups. In laparoscopic group, two 5 and 10 mm portals were inserted; First in the umbilicus for introducing the camera and the second, caudal to the umbilicus for inserting the forceps. Laparoscopic procedure involved grasping and tacking the ovary to the abdominal wall, followed by electrocautery, resection and removal of the ovary. In open surgery, routine OVE was conducted through an incision from umbilicus to caudal midline. Mean operative time, total length of scar, blood loss, clinical and blood parameters and all intra and post-operative complications were recorded in both groups. Mean operative time, total length of scar, blood loss and post-operative adhesions were significantly less in laparoscopic group compared with open surgery. In conclusion, laparoscopic OVE is an acceptable procedure due to more advantages in comparison with traditional OVE.

Heat-induced conformational unfolding of fatty acid-free and fatted camel serum albumin: a comparative study.

Albumin is a multifunctional non-glycosylated, negatively charged plasma protein, with extraordinary ligand-binding and transport properties, antioxidant functions, and enzymatic activities. Physiologically, albumin transports free fatty acids in plasma and contributes in maintaining colloid osmotic pressure. Recent progresses in using albumin as a versatile protein carrier for drug targeting and for improving the pharmacokinetic profile of peptide or protein-based drugs, increased the attempts for improving albumin stability. Studying the thermal stability of camel albumin may provide us not only new clues for designing recombinant albumins, but also molecular insights on camel physiology. This study aims to determine the thermal stability of camel albumin. Fatted camel serum albumin (FCSA) was purified from blood via combination of Cohn's method and anion-exchange chromatography. Activated charcoal treatment was used to obtain defatted camel serum albumin (CSA). Fluorescence spectroscopy and differential scanning calorimetry (DSC) were used to study thermal denaturation of this protein. The set of fluorescence spectra were deconvoluted using the convex constraint analysis method (CCA). The results from deconvolution of fluorescence spectroscopy and DSC showed three and two components for CSA and FCSA, respectively. The bimodal DSC transition can be attributed to a crevice between domains I and II and formation of two independent thermodynamic domains. The crevice formation can be prevented by fatty acid binding between domains I and II. The calculated values of ∆H v/∆H cal, approximately 0.4 for CSA and near 1 for FCSA, confirmed the presence of at least one intermediate in thermal unfolding of CSA and the absence of the intermediate for FCSA. The obtained midpoint transition temperature (T m) of FCSA was about 20 °C higher than that of CSA. Such enormous stabilizing effect may be attributed to the fact that fatty acid serves as glue which preserves different domains beside each other and prevents formation of the mentioned intermediate.

Comparative evaluation of two reconstructive methods following laparoscopic assisted subtotal gastrectomy in dogs.

BACKGROUND: Laparoscopic gastrectomy is a new and technically challenging surgical procedure with potential benefit. The objective of this study was to investigate clinical and para-clinical consequences following Roux-en-Y and Jejunal Loop interposition reconstructive techniques for subtotal gastrectomy using laparoscopic assisted surgery. RESULTS: Following resection of the stomach attachments through a laparoscopic approach, stomach was removed and reconstruction was performed with either standard Roux-en-Y (n = 5) or Jejunal Loop interposition (n = 5) methods. Weight changes were monitored on a daily basis and blood samples were collected on Days 0, 7 and 21 post surgery. A fecal sample was collected on Day 28 after surgery to evaluate fat content. One month post surgery, positive contrast radiography was conducted at 5, 10, 20, 40, 60 and 90 minutes after oral administration of barium sulfate, to evaluate the postoperative complications. There was a gradual decline in body weight in both experimental groups after surgery (P < 0.05). There was no difference in blood parameters at any time after surgery between the two methods (P > 0.05). Fecal fat content increased in the Roux-en-Y compared to the Jejunal loop interposition technique (P < 0.05). No major complications were found in radiographs and gastric emptying time was similar between the two groups (P > 0.05). CONCLUSION: Roux-en-Y and Jejunal loop interposition techniques might be considered as suitable approaches for reconstructing gastro-intestinal tract following gastrectomy in dogs. The results of this study warrant further investigation with a larger number of animals.

Comparison between single and three portal laparoscopic splenectomy in dogs.

BACKGROUND: Single incision laparoscopic surgery (SILS) is a newly growing technique to replace a more invasive conventional multiple portal laparoscopic surgery. The objective of this study was to compare single (SILS) with three portal (Conventional) laparoscopic splenectomy in dogs. Mongrel dogs (n = 18), weighting 15 ± 3 kg, were selected for this study (n = 12 SILS; n = 6 conventional). The area from xiphoid to pubis was prepared under aseptic conditions in dorsal recumbency with the head down and tilted 30 degree in the right lateral position. Pneumoperitoneum was established by CO2 using an automatic high flow pressure until achieving 12 mm Hg. Instrumentation used consisted of curved flexible-tip 5 mm Maryland forceps and ultracision harmonic scalpel for sealing and cutting of the vessels and splenic attachments. RESULTS: All dogs recovered uneventfully. The splenectomy procedure using SILS and conventional methods were significantly different in the respective operative time (29.1 ± 1.65 vs. 42.0 + 2.69 min) and the length of the surgical scar (51.6 ± 1.34 mm vs. 72.0 ± 1.63 mm; P < 0.001). There were no post-operative wound complication including inflammation, infection, hernia formation and dehiscence up to one month after surgery. Meanwhile, the conversion to open surgery or application of additional portals was not required in both approaches. CONCLUSION: This study demonstrated that SILS is a safe and feasible operation and could be used as an alternative approach to three portal (Conventional) for splenectomy in dog.

Comparative evaluation of conventional and transvaginal laparoscopic ovariohysterectomy in dogs.

OBJECTIVE: To evaluate the feasibility and safety of a transvaginal approach for laparoscopic ovariohysterectomy (OVH) in dogs and to compare it with conventional laparoscopic OVH. STUDY DESIGN: Prospective study. ANIMALS: Adult healthy female mixed breed dogs (n = 24). METHODS: Dogs (weighing 14-17 kg) were anesthetized and positioned in dorsal recumbency for ovariohysterectomy. Dogs were prepared for either conventional (n = 12) or transvaginal (n = 12) laparoscopic OVH. For conventional laparoscopic OVH, 3 midline abdominal portals were used and for the transvaginal approach, 2 midline abdominal portals and one vaginal portal were used. The transected ovarian pedicles, broad ligament, and uterus were removed through the umbilical region in the conventional method and through the vagina in the transvaginal method. Mean surgical time, intraoperative and postoperative complications, clinical and hematologic findings, and wound complications were compared. RESULTS: OVH was successfully performed without complications using both methods. Mean ± SD surgical times were similar between conventional (34.2 ± 4.03 min) and transvaginal (37.0 ± 3.56 min) methods. No significant differences, hematologic and clinical variables, were found between groups. The vaginal port could limit surgical maneuvers ergonomically during manipulation of the uterine body. CONCLUSIONS: Transvaginal approach for laparoscopic OVH is a feasible technique with the advantage of requiring one less abdominal portal.

Biological activity of camel milk casein following enzymatic digestion.

The aim of this study was to investigate the effects of enzymatic hydrolysis with digestive enzymes of camel whole casein and beta-casein (ß-CN) on their antioxidant and Angiotensin Converting Enzyme (ACE)-inhibitory properties. Peptides in each hydrolysate were fractionated with ultra-filtration membranes. The antioxidant activity was determined using a Trolox equivalent antioxidant capacity (TEAC) scale. After enzymatic hydrolysis, both antioxidant and ACE-inhibitory activities of camel whole casein and camel ß-CN were enhanced. Camel whole casein and ß-CN showed significant ACE-inhibitory activities after hydrolysis with pepsin alone and after pepsinolysis followed by trypsinolysis and chymotrypsinolysis. Camel ß-CN showed high antioxidant activity after hydrolysis with chymotrypsin. The results of this study suggest that when camel milk is consumed and digested, the produced peptides start to act as natural antioxidants and ACE-inhibitors.

Structure and stability analysis of cytotoxic complex of camel α-lactalbumin and unsaturated fatty acids produced at high temperature.

α-Lactalbumin α-La), together with oleic acid can be converted to a complex, which kills tumor cells selectively. Cytotoxic α-La -oleic acid and α-La -linoleic acid complexes were generated by adding fatty acid to camel holo α-La at 60 ° C (referred to as La-OA-60 and La-LA-60 state, respectively). Structural properties of these complexes were studied and compared to the camel α-La. The experimental results show that linoleic acid induces α-La partial unfolding but oleic acid does not change the protein structure significantly. Also the stability of La-OA-60 and La-LA-60 toward thermal denaturation was measured. The order of temperature at the transition midpoint is as follows: La-LA-60 < La-OA-60 < α-La. La-OA-60 complex inhibited tubulin polymerization in vitro. Although the structures of La-OA-60 and La-LA-60 were different, these two complexes had similar cytotoxic effect to DU145 human prostate cancer cells. Samples of La-OA-60 that have been renatured after denaturation lost the specific biological activity toward tumor cells.

Endoscopic treatment of vesicoureteral reflux using calcium hydroxyl apatite in dogs.

BACKGROUND: Injection of biomaterial to suburetral region, using minimally invasive procedure, has become an interesting topic for urologists to treat vesicoureteral reflux. The objective of this study was to evaluate the feasibility of injecting newly introduced calcium hydroxyl apatite to suburetral region, for treating an experimentally induced vesicoureteral reflux in dogs. FINDINGS: Bilateral vesicoureteral refluxed (VUR) mixed breed dogs (n = 12; 10-15 kg live weight, 3-6 months of age) were selected for this study. The presence and grade of the reflux were determined using cystography. Accordingly, 6 dogs displayed grade 1 & 2 and the other 6 showed grade 3 & 4 bilateral VUR. Every single dog, with bilateral VUR, underwent endoscopic treatment and received an injection of calcium hydroxyl apatite (an Iranian made product) into the left (treated side) and an injection of the similar volume of normal saline in to the right (control side) subureteric space. One week, 3 and 6 months after treatment, cystography was performed. On each occasion, 4 dogs were euthanized by gas inhalation and biopsy samples were collected for histopathological study from ureter, bladder, kidney, lung and spleen in order to investigate the biomaterial migration into different organs. Data were analyzed using Chi-squared test. In control sides, radiographs confirmed the same grade of VUR, found at the initiation of the study. VUR was resolved in 100% (6/6) of Grade 1 & 2 and 83.33% (5/6) of Grade 3 & 4 in treated side. Therefore, the total success rate of this study was 91.67% (11/12). Macroscopic examination of the vesicouretral region of the treated side revealed a firm and consistent biomaterial mass at the site of injection. Histological findings confirmed inflammation at treated side. In contrast, there was no tissue reaction on control side. There was no evidence for biomaterial migration in macroscopic and microscopic observations in this study. CONCLUSION: In the present study, a new biocompatible material produced a firm, consist and sustainable biomaterial mass in the suburetral region for treating vesicouretral reflux without any evidence of biomaterial migration.

Effect of fat supplementation during transition period on plasma leptin and non-esterified fatty acid concentrations in Holstein cows.

The objective of this experiment was to investigate the effect of fat supplementation during the transition period on pre and postpartum body weight (BW), body condition score (BCS), non-esterified fatty acids (NEFA), glucose and leptin concentrations in Holstein cows. Holstein cows (n = 15) received a low fat diet (LF; 1.61 Mcal net energy for lactation (NEL)/kg of dry matter [DM]), moderate fat diet (MF; 1.68 Mcal NEL/kg DM) or a high fat diet (HF; 1.74 Mcal NEL/kg DM) for 4 weeks prior to calving. All cows were fed similar lactation diets ad libitum (1.74 Mcal NEL/kg DM) for 30 days after calving. Increasing diet energy density during transition period had no effect on prepartum DMI, BCS, BW, glucose and NEFA concentrations (P > 0.05); but leptin concentrations and energy balance (EB) were affected by treatments (P < 0.05). Animals fed HF had less plasma leptin prepartum. After parturition, BW, milk production, milk fat, protein, urea nitrogen and plasma glucose concentrations were affected by prepartum diets (P < 0.05). Fat supplementation prepartum did not affect postpartum NEFA. In conclusion, prepartum fat supplementation decreased leptin concentration prepartum.

Improvement of the antimicrobial and antioxidant activities of camel and bovine whey proteins by limited proteolysis.

The compositions and structures of bovine and camel milk proteins are different, which define their functional and biological properties. The aim of this study was to investigate the effects of enzymatic hydrolysis of camel and bovine whey proteins (WPs) on their antioxidant and antimicrobial properties. After enzymatic treatment, both the antioxidant and the antimicrobial activities of bovine and camel WPs were improved. The significantly higher antioxidant activity of camel WPs and their hydrolysates as compared with that of bovine WPs and their hydrolysates may result from the differences in amounts and/or in accessibilities of antioxidant amino acid residues present in their primary structures and from the prevalence of alpha-lactalbumin and beta-lactoglobulin as proteolytic substrates in camel and bovine whey, respectively. The results of this study reveal differences in antimicrobial and antioxidant activities between WP hydrolysates of bovine and camel milk and the effects of limited proteolysis on these activities.