Estimation of periostin as a biomarker for early pregnancy diagnosis in goats: a preliminary study.

BACKGROUND: Periostin (POSTN) is an extracellular matrix (ECM) protein that plays an important role in the metastatic process and cancer cell migration. As implantation is a similar mechanism to metastasis, it has been hypothesized that POSTN may also play a role in the implantation process. AIMS: The aim of the present study was to compare POSTN and progesterone levels during the early pregnancy stage in Damascus goats. METHODS: Forty goats were synchronized using progesterone based sponges and were mated upon estrus signs display. While ten goats were kept as control (CON) and were not allowed to mate. Blood samples were taken through jugular venepuncture from CON and synchronized goats on day 13, 15, 17, 19, and 21 of breeding. Progesterone and POSTN levels were determined by enzyme-linked immunosorbent assay (ELISA). Later the pregnancy diagnosis was confirmed by transabdominal ultrasonography on day 50 after mating. RESULTS: Progesterone level was influenced by status of pregnancy and day of observation with an interaction between the status of pregnancy and day of observation in goats. Whereas POSTN level was only affected by the day of observation. CONCLUSION: POSTN level did not vary with progesterone level during phase of embryonic implantation in goats; however, standardization and application of different procedures for POSTN assay in a large group of animals might be useful as an early pregnancy biomarker in goats.

Variations in serum metabolites in response to season, cyclicity, and pregnancy in estrus-synchronized Damascus goats.

The current study was carried out to determine the changes in serum electrolytes, liver enzymes, and hormones during breeding and non-breeding seasons in estrus-synchronized goats. Forty goats (n = 40) were synchronized by inserting the sponges intravaginally for 12 days; PMSG and PGF2α were injected on day of sponge removal. Blood samples were collected from the goats at the time of sponges insertion (day -12), estrus day (day 0), and at 15 day after sponge withdrawal (day 15). The blood samples were analyzed for glucose, cholesterol (CHO), aspartate aminotransferase (AST), alanine aminotransferase (ALT), thyroid stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4) and progesterone (P4), calcium (Ca), phosphate (PO4), magnesium (Mg), sodium (Na), potassium (K), and chlorine (Cl) levels. Repeated measurements ANOVA of general linear model were used as statistical model. The results showed that seasons have an effect (p < 0.05) on glucose, Ca, Na, K, Cl, CHO, T3, and T4 levels. There was an effect (p < 0.05) of cyclicity only on P4 level. Whereas, nonpregnant, or goats carrying singlet and twin fetuses have shown variation (p < 0.05) in glucose, Ca, and P4 levels. Although, the correlations existed between different serum metabolites but strong correlations (p < 0.05) were observed either between Ca and Na or T3 and T4. In conclusion, the serum electrolytes and liver enzymes are correlated with hormones during the different seasons, pregnancy status, and fecundity. In recommendation, the results of the study could be as managemental tool to monitor the reproductive activity across seasons and to maintain pregnancy carrying twining in goat breeds.

Effects of folic acid on testicular toxicity induced by bisphenol-A in male Wistar rats.

We investigated the protective effect of the folic acid (FA) against bisphenol-A (BPA) induced toxicity in rat testis. We used four groups of seven adult male Wistar albino rats. The control group was fed corn oil, the BPA group was given BPA, the FA group was given FA and the FA + BPA group was given FA initially followed by BPA 1 h later. The BPA, FA and corn oil were administered by oral gavage for 14 days. At the end of the experiment, testis sections were examined for histological and histomorphometric characteristics. The TUNEL method was used to detect apoptosis and immunohistochemistry was used to examine the distribution of spermatogonial stem cells. Levels of serum testosterone were measured, and sperm viability and morphology were determined. The histological structure of the testis was normal in the control and FA groups. Although the number of TUNEL positive cells/tubule increased, the seminiferous epithelium height (SEH) at stages VII-VIII decreased in the BPA group compared to the control, FA and FA + BPA groups. The number of TUNEL positive cells/tubule decreased and the SEH at stages VII-VIII increased in the FA + BPA group compared to the BPA group. No significant difference in spermatogonial stem cells was found among groups. The level of serum testosterone and percentage of viable sperm was significantly lower, while the head, midpiece and total sperm abnormalities were significantly higher in the BPA treated group compared to control, FA, FA + BPA groups. It appears that the toxic effects of BPA on testis might be minimized by FA treatment.

Comparative efficacy of G6G and Ovsynch protocols on synchronization and pregnancy rate in Nili-Ravi buffalo.

The aim of this work was to evaluate the efficacy of G6G and Ovsynch protocols on synchronization and pregnancy rate in Nili-Ravi buffalo. For this, a total of fifty buffalo were assigned into two treatment groups: (1) G6G (n=25) and (2) Ovsynch (n=25). The blood samples were collected from all the treated buffalo on day (D) 7 (PGF2α injection of Ovsynch) to measure the progesterone (P4). All the buffalo were fixed time inseminated (FTAI) following last GnRH of Ovsynch. Ultrasonography was performed for follicular, corpus luteum (CL) dynamics and pregnancy diagnosis. The results revealed that synchronization rate and response to final PGF2α were significantly (P<0.05) higher in G6G compared to Ovsynch. The ovulation to first GnRH of Ovsynch was higher (P<0.05) in G6G (84%) than Ovsynch (56%). The mean dominant follicle size following last GnRH of Ovsynch showed significantly larger diameter in G6G (11.7±0.2mm) compared to Ovsynch (10.9±0.2mm). Moreover, the higher (P<0.05) P4 concentration was observed in G6G than Ovsynch. The presence of double CL on D 7 was higher (P<0.05) in G6G (64%) than Ovsynch (24%) group. The pregnancy rate (P=0.08) tended to be greater in G6G compared to Ovsynch. In conclusion, G6G treatment showed better ovulatory and luteolytic response to first GnRH and PGF2α compared to Ovsynch treatment. Moreover, the addition of GnRH and PGF2α prior to Ovsynch is helpful to improve synchronization application of Ovsynch protocol in Nili-Ravi buffalo.

Trehalose enhances osmotic tolerance and suppresses lysophosphatidylcholine-induced acrosome reaction in ram spermatozoon.

This study was aimed to investigate the influence of trehalose on osmotic tolerance and the ability of ram spermatozoon to undergo acrosome reaction induced by lysophosphatidylcholine (LPC). In experiment 1, the diluted ejaculates were exposed to anisosmotic fructose solutions (70, 500, 750 and 1000 mOsm l(-1) ) with or without 50 mm trehalose. The presence of trehalose in hyperosmotic conditions enhanced (P < 0.05) the percentage of live, live-intact and intact spermatozoa. Similarly, trehalose enhanced (P < 0.05) the live and live-intact spermatozoa during hypo-osmotic conditions. In experiment 2, the centrifuged ejaculates were diluted with TCG only or TCG containing either 50 or 100 mm trehalose. The acrosome reaction was induced by LPC. The percentage of acrosome-reacted spermatozoon was less (P < 0.05) in trehalose-supplemented groups compared to control. In experiment 3, the ejaculates were cryopreserved in an extender containing 0 mm (control), 50 mm or 100 mm trehalose. Supplementation of extender with trehalose, either 50 mm or 100 mm, enhanced the cryosurvival rate (P < 0.05) compared to the control. In conclusion, the presence of trehalose in anisosmotic conditions enhances the osmotic tolerance, cryosurvival rate of ram spermatozoon and suppresses their ability to undergo LPC and cryo-induced acrosome reaction.

Efficacy of Retigabine on Acute Limbic Seizures in Adult Rats.

BACKGROUND AND PURPOSE: The efficacy of retigabine (RGB), a positive allosteric modulator of K+ channels indicated for adjunct treatment of partial seizures, was studied in two adult models of kainic acid (KA)-induced status epilepticus to determine it's toleratbility. METHODS: Retigabine was administered systemiclly at high (5 mg/kg) and low (1-2 mg/kg) doses either 30 min prior to or 2 hr after KA-induced status epilepticus. High (1 µg/µL) and low (0.25 µg/µL) concentrations of RGB were also delivered by intrahippocampal microinjection in the presence of KA. RESULTS: Dose-dependent effects of RGB were observed with both models. Lower doses increased seizure behavior latency and reduced the number of single spikes and synchronized burst events in the electroencephalogram (EEG). Higher doses worsened seizure behavior, produced severe ataxia, and increased spiking activity. Animals treated with RGB that were resistant to seizures did not exhibit significant injury or loss in GluR1 expression; however if stage 5-6 seizures were reached, typical hippocampal injury and depletion of GluR1 subunit protein in vulernable pyramidal fields occurred. CONCLUSIONS: RGB was neuroprotective only if seizures were significantly attenuated. GluR1 was simultaneously suppressed in the resistant granule cell layer in presence of RGB which may weaken excitatory transmission. Biphasic effects observed herein suggest that the human dosage must be carefully scrutinized to produce the optimal clinical response.

Retigabine calms seizure-induced behavior following status epilepticus.

In adult rats, intraperitoneal injection of kainate (KA) results in sustained status epilepticus and persistent behavioral comorbidities such as hyperexcitability, anxiety, and altered response to environmental cues. Intrahippocampal KA also results in sustained status epilepticus and continuous high frequency oscillations in the electroencephalograph (EEG), although subsequent behavioral side effects are unknown. We hypothesized that retigabine, a recently discovered anticonvulsant and potent positive modulator of Kv7 channels, may attenuate seizure-induced behavioral abnormalities. Status epilepticus was induced by administration of KA either intraperitoneally (15 mg/kg) or by single intrahippocampal injection (1.0 µg/0.5 µL). After 24 h, half of systemically KA-treated animals that reached stage 6 seizures were injected once daily with retigabine (5 mg/kg) for 14 continuous days. All groups underwent three behavioral tests--capture and handling, open field, and elevated plus maze--24 h following the last retigabine treatment and were sacrificed at 25-28 days. In the capture and handling test, systemic KA treatment resulted in frisky behavior and resistance to capture with wild attempts to escape during the 1st, 2nd, and 3rd weeks of the observation period. In contrast, these behaviors were attenuated in KA+retigabine-treated animals. In the open-field test, KA-treated animals spent more time in the center zone, but KA+retigabine-treated rats had greater overall activity compared with those having vehicle, KA, or retigabine-only treatment. In the elevated plus maze, KA+retigabine-treated animals traveled greater distances in open and closed arms (proximal and distal) compared with controls, also signifying anxiety reduction. Retigabine-only-treated rats traveled more in the open proximal arms compared with controls, indicating increased hyperlocomotion in normotensive rats. Although treatment with KA+retigabine resulted in anxiolytic-like effects in all three behavioral tasks compared with vehicle, this group did not significantly differ from systemically KA-treated rats in most measurements in open-field and elevated plus maze tasks, suggesting that retigabine may also cause hyperlocomotion unrelated to anxiety level. Despite that intrahippocampal KA-treated rats displayed comparable seizure behavior, epileptiform activity, and hippocampal injury, their behavior resembled the controls, suggesting that molecular and subsequent cellular changes are also partially responsible for anxiolytic-like effects and that these results are likely independent of the hippocampus.

Fertility following CIDR based synchronization regimens in anoestrous Nili-Ravi buffaloes.

The objective of this study was to compare oestrus expression and fertility rate in used and new controlled internal drug releasing (CIDR) device treated anoestrous buffaloes. Furthermore, to determine the timing of ovulation, and fertility rate in estradiol benzoate (EB) and GnRH-administered CIDR-treated anoestrous Nili-Ravi buffaloes. In experiment 1, buffaloes received either a used CIDR (UCIDR, n = 35) or a new CIDR (NCIDR, n = 36) for 7 day and PGF2α on day 6. Oestrous expression was similar (p > 0.05) between UCIDR (88.5%) and NCIDR (96.6%) buffaloes. The pregnancy rate did not differ (p > 0.05) because of treatment (37.1% in UCIDR vs 36.6% in NCIDR). In experiment 2, buffaloes (n = 55) received CIDR device for 7 days and PGF2α, on day 6 and randomly assigned into three treatment groups: (i) CIDR-EB (n = 17) received EB on day 8, (ii) CIDR-GnRH (n = 18) received GnRH on day 9 and (iii) control (n = 20) received no further treatment. Mean interval from CIDR removal to ovulation in CIDR-EB, CIDR-GnRH and CIDR group were 61.3 ± 0.8, 64.9 ± 1.8 and 65.1 ± 16.7 h, respectively. However, the buffaloes in the CIDR-EB and CIDR-GnRH group had lesser variability in the timing of ovulation compared to control. The pregnancy rate of both CIDR-EB group (58%) and CIDR-GnRH group (61%) were tended to be higher (p < 0.1) than control (30%). In conclusion, compared to NCIDR devices, previously UCIDR devices are equally effective to induce oestrus in anoestrous buffaloes resulting optimal pregnancy rate. Administration of EB and GnRH after CIDR removal results in tighter synchrony (less variability) and improved fertility in anoestrous buffaloes. CIDR based synchronization regimens have great potential in fertility improvement in anoestrous buffaloes.

Lymph node retrieval and assessment in stage II colorectal cancer: a population-based study.

BACKGROUND: Adjuvant chemotherapy for patients with stage III (node-positive) colorectal cancer (CRC) reduces mortality by one third. Retrieval of an inadequate number of lymph nodes in the surgical specimen may result in incorrectly designating some patients as stage II (node negative), and consequently, such patients may not be offered appropriate chemotherapy. Recent National Cancer Institute guidelines suggest that a minimum of 12 nodes should be examined to ensure accurate staging. METHODS: This population-based study identified stage II (T3N0 and T4N0) CRC cases by using CRC pathology reports (1997-2000) from the Ontario Cancer Registry. Patients aged 19 to 75 years were identified, and demographic, surgical, pathologic, and hospital data were extracted. Factors relating to the number of lymph nodes assessed were examined. RESULTS: A total of 8848 CRC cases were reviewed, and 1789 stage II cases were identified. Seventy-three percent of cases were designated as node negative on the basis of assessment of <12 lymph nodes. Multivariate analysis showed that age, tumor size, specimen length, use of a pathology template, and academic status of the hospital were significant predictors of the number of lymph nodes assessed. CONCLUSIONS: A subset of patients with CRC in Ontario were assigned stage II disease on the basis of examination of relatively few lymph nodes.