In vitro maturation medium supplementation: utilization of repaglinide, L-carnitine, and mesenchymal stem cell-conditioned medium to improve developmental competence of oocytes derived from endometriosis mouse models.

Endometriosis (EMS) is one of the most prevalent causes for female infertility. Herein, we investigated the effect of the repaglinide (RG), L-carnitine (LC), and bone marrow mesenchymal stem cell-conditioned medium (BMSC-CM) supplementation during in vitro maturation (IVM) on the quality, maturation, and fertilization rates, as well as embryonic quality and development of oocytes derived from normal and EMS mouse model. Immature oocytes were collected from two groups of normal and EMS-induced female NMRI mice at 6-8 weeks of age. Oocytes were cultured in IVM medium unsupplemented (control group), or supplemented with 1 M RG, 0.3 and 0.6 mg/mL LC, and 25 and 50% BMSC-CM. After 24 h of oocyte incubation, IVM rate and antioxidant status were assessed. Subsequently, the rates of fertilization, cleavage, blastulation, and embryonic development were assessed. Our results demonstrated that supplementation of IVM medium with LC and BMSC-CM, especially 50% BMSC-CM, significantly enhanced IVM and fertilization rates, and markedly improved blastocyst development and total blastocyst cell numbers in EMS-induced mice compared to the control group (53.28±0.24 vs 18.09±0.10%). Additionally, LC and BMSC-CM were able to significantly modulate EMS-induced nitro-oxidative stress by boosting total antioxidant capacity (TAC) and mitigating nitric oxide (NO) levels. Collectively, LC and BMSC-CM supplementation improved oocyte quality and IVM rates, pre-implantation developmental competence of oocytes after in vitro fertilization, and enhanced total blastocyst cell numbers probably by attenuating nitro-oxidative stress and accelerating nuclear maturation of oocytes. These outcomes may provide novel approaches to refining the IVM conditions that can advance the efficiency of assisted reproductive technologies in infertile couples.

In vitro maturation medium supplementation: utilization of repaglinide, L-carnitine, and mesenchymal stem cell-conditioned medium to improve developmental competence of oocytes derived from endometriosis mouse models

Endometriosis (EMS) is one of the most prevalent causes for female infertility. Herein, we investigated the effect of the repaglinide (RG), L-carnitine (LC), and bone marrow mesenchymal stem cell-conditioned medium (BMSC-CM) supplementation during in vitro maturation (IVM) on the quality, maturation, and fertilization rates, as well as embryonic quality and development of oocytes derived from normal and EMS mouse model. Immature oocytes were collected from two groups of normal and EMS-induced female NMRI mice at 6-8 weeks of age. Oocytes were cultured in IVM medium unsupplemented (control group), or supplemented with 1 M RG, 0.3 and 0.6 mg/mL LC, and 25 and 50% BMSC-CM. After 24 h of oocyte incubation, IVM rate and antioxidant status were assessed. Subsequently, the rates of fertilization, cleavage, blastulation, and embryonic development were assessed. Our results demonstrated that supplementation of IVM medium with LC and BMSC-CM, especially 50% BMSC-CM, significantly enhanced IVM and fertilization rates, and markedly improved blastocyst development and total blastocyst cell numbers in EMS-induced mice compared to the control group (53.28±0.24 vs 18.09±0.10%). Additionally, LC and BMSC-CM were able to significantly modulate EMS-induced nitro-oxidative stress by boosting total antioxidant capacity (TAC) and mitigating nitric oxide (NO) levels. Collectively, LC and BMSC-CM supplementation improved oocyte quality and IVM rates, pre-implantation developmental competence of oocytes after in vitro fertilization, and enhanced total blastocyst cell numbers probably by attenuating nitro-oxidative stress and accelerating nuclear maturation of oocytes. These outcomes may provide novel approaches to refining the IVM conditions that can advance the efficiency of assisted reproductive technologies in infertile couples.

Moderate overfeeding of different sources of metabolizable energy and protein. II: Effects on inflammatory status of sheep in late pregnancy and growth trajectory of the offspring.

Our objective was to evaluate the effects of moderate overfeeding of different sources of metabolizable energy (ME) and protein (MP) on inflammatory status of sheep in late gestation and short-term growth performance of the offspring. A total of 32 healthy pregnant Zell ewes (2.5 ± 1 years old, 42.19 ± 1.65 kg body weight [BW]) were selected 7 weeks before lambing. After receiving a ten-day adaptation diet, eight ewes were randomly allocated to each of the four treatments including: 1) The adaptation diet that fulfills ME and MP requirements, according to the National Research Council's (NRC, 2007) recommendations for late pregnant ewes (CON); 2) A diet with 10% higher levels of ME than CON, using corn grain for extra ME (HEC); 3) A diet with 10% higher level of ME compared to CON, using flaxseed for additional ME (HEF); and 4) A diet with a 10% higher level of MP than CON, using a rumen-protected lysin and methionine supplement (RP-Lys & Met) to supply extra MP (PRO). After lambing, ewes continued their dietary treatments until 30 days postpartum; however, the content of ingredients were proportionally increased to meet the recommended levels of energy and protein for lactating ewes (NRC, 2007). Blood samples from the ewes were taken on days -21, -14, and -7 before parturition. Colostrum yield and components were determined, and lamb's BW and milk intake (MI) were measured weekly until 28 days of age. Other growth performance indexes were computed accordingly. Blood samples from the offspring were taken on days 15 and 30 of life. The data was analyzed using SAS software (SAS Institute Inc., 9.4). Compared with the CON, HEF-fed ewes were the only group that had lower levels of oxidative stress and blood inflammatory markers at -7 d before parturition (P < 0.05). Concentrations of aminotransferases in CON-fed ewes were greater than other groups in late gestation (P ≤ 0.05). Offspring of HEF- and PRO-fed ewes had higher birth BW than lambs of the CON-fed group (3.71 and 4.11 vs. 3.01 ± 0.24, Kg for HEF, PRO, and CON, respectively). Except for week 2, lambs of CON-fed ewes had lower BW gain compared with the other offspring (P < 0.01). Lambs of PRO-fed ewes had greater levels of plasma immunoglobulin G (IgG, days 15 and 30 of life; P < 0.05) and MI (from week 2; P < 0.001) than other groups. In conclusion, regardless of the source, moderate overfeeding of ME/MP was more beneficial for the immune status of late pregnant dams and the offspring's performance than non-overfed ones.

The Antioxidant Effect of Medicago sativa L. (Alfalfa) Ethanolic Extract against Mercury Chloride (HgCl2) Toxicity in Rat Liver and Kidney: An In Vitro and In Vivo Study.

Heavy metals such as mercury are some of the environmental pollutants and can induce toxicity by bioaccumulation and oxidative damage. This study aimed to investigate the effect of ethanolic extract of Medicago sativa L. (Alfalfa) on mercury damage in the kidney and liver of rats. Thirty Wistar rats were randomly divided into five groups, the control group, S group (2 mg/kg mercury chloride), and T1, T2, and T3 groups that, in addition to mercury, received doses of 250, 500, and 750 mg/kg of the alfalfa extract. On the last day, blood samples were taken, and the serum was separated to measure biochemical and oxidative stress parameters in the kidney and liver. A part of the kidney and liver was also used for histopathological evaluation. Total phenols and flavonoids were 40.45 ± 2.12 and 14.36 ± 0.45 mg/g, respectively, whereas IC50 was 245.18 ± 19.76 µg/ml. The body weight significantly decreased in the S group compared to other groups, while treatment with different doses of alfalfa extract increased the body weight. Mercury concentration in the kidney was higher than that in the liver. The serum levels of urea, creatinine, alanine aminotransferase (ALT), and alkaline phosphatase (ALP) significantly increased in the S group compared to the control group, while treatment with different doses of alfalfa extract increased their levels. Moreover, an increase in malondialdehyde (MDA) and a decrease in glutathione peroxidase (GPx), catalase (CAT), total antioxidant capacity (TAC), and superoxide dismutase (SOD) activity were observed in the S group. The level of these parameters significantly improved in the groups receiving the extract compared to the S group. Furthermore, the histopathological evaluation showed glomerular and tubular damage and hepatic necrosis in the S group and that these conditions improved in the T3 group. The findings of this study showed that the ethanolic extract of alfalfa in a dose-dependent manner has potentially unique protective effects against mercury poisoning in the kidney and liver.

De-oiled soy lecithin positively influenced growth performance, nutrient digestibility, histological intestinal alteration, and antioxidant status in turkeys fed with low energy diets.

1. The purpose of this research was to investigate the effects of supplementing an emulsifier (de-oiled soybean lecithin (DSL)) in a low metabolisable energy (ME) diet on growth performance, nutrient digestibility, carcase characteristics, intestinal morphology, blood metabolites, and antioxidant status in growing turkeys.2. A total of 480 one-day-old turkeys were assigned to one of four dietary treatments with of eight replicates of 15 birds each. Experimental treatments included a basal diet (BE) with commercially recommended levels of ME, a reduced energy diet (RE) with 0.42 MJ/kg reduction in dietary ME content, the RE diet + 1 g/kg DSL (DSL-1), and RE + 2 g/kg DSL (DSL-2).3. After 112 days, the body weight, average daily gain, and feed:gain in turkeys fed the supplemented for BE diets were better (P < 0.05) than in those fed RE, and those fed diet DSL-2 had the best performance. Although the RE diet decreased abdominal fat and relative liver weight (P < 0.05), compared to the BE diet, and supplementation with either level of DSL did not influence these variables.4. There were linear increases (P < 0.05) in fat digestibility, nitrogen-corrected apparent ME, and duodenal villus height, villus height/crypt depth ratio, and villus surface area in LE diet supplemented with DSL. From the jejunal morphology, crypt depth was decreased by DSL-supplemented diets (P < 0.05).5. Serum triglyceride, total cholesterol, and malondialdehyde concentrations were lower, whereas the serum superoxide dismutase activity was greater for the DSL-2 group compared to the BE and RE groups (P < 0.05).6. The findings suggested that, while low-ME diets impaired turkey growth performance, dietary supplementation of DSL could reverse such impacts of these diets. The DSL-supplemented diet at the inclusion level of 2 g/kg was advantageous over both BE and RE diets in terms of intestinal morphology, lipid profile, and antioxidant status in growing turkeys.

Pentoxifylline besides naltrexone recovers morphine-induced inflammation in male reproductive system of rats by regulating Toll-like receptor pathway.

This study aimed to investigate the effect of pentoxifylline on complications of prolonged usage of morphine upon the testis and sperm parameters of rats. In this study, forty male Wistar rats were divided into five groups (n = 8) and treated for 56 days to only saline, only morphine, only pentoxifylline, pentoxifylline + morphine and naltrexone + morphine. The diameters of seminiferous tubules, the maturity of germ line epithelium and sperm parameters were evaluated. The expression of inflammatory-related factors in testis tissues were also investigated at gene and protein levels. The data were calculated by one-way ANOVA test followed by Tukey's post hoc test using SPSS software for windows (version 20). Seminiferous tubule diameter, the maturity of spermatogonia and sperm parameters were significantly decreased in morphine group in comparison with control, pentoxifylline and pentoxifylline + morphine groups (p < .001). The expression of anti-inflammatory markers, at both gene and protein levels, was significantly increased in testis of morphine-treated rats in comparison with other groups (p < .001). Chronic morphine administration induces destructive effects on male reproductive system by regulating inflammatory responses. Pentoxifylline recovers the destructive effects of morphine on male reproductive system by inhibiting TLR (Toll-like receptor) activity, as an anti-inflammatory response.

Morphine-induced conditioned place preference and the alterations of p-ERK, p-CREB and c-fos levels in hypothalamus and hippocampus: the effects of physical stress.

The hypothalamus and hippocampus are important areas involved in stress responses and reward processing. In addition, ERK/CREB pathway plays a critical role in the control of cellular responses to stress and reward. In the current study, effects of acute and subchronic stress on the alteration of p-ERK, p-CREB and c-fos levels in the hypothalamus and hippocampus of saline- or morphine-treated animals during morphine-induced conditioned place preference (CPP) procedure were investigated. Male Wistar rats were divided into two saline- and morphine-treated supergroups. Each supergroup includes of control, acute stress and subchronic stress groups. In all of groups, the CPP procedure was done, afterward the alternation of p-ERK/ERK ratio, p-CREB/CREB ratio and c-fos level in the hypothalamus and hippocampus were estimated by Western blot analysis. The results indicated that in saline- or morphine-treated animals, p-ERK/ERK ratio, p-CREB/CREB ratio and c-fos level increased after application of acute and subchronic stress (except for p-ERK/ERK ratio in morphine-control group). Our findings revealed that in saline- or morphine-treated animals, acute and subcronic stress increased the p-ERK/ERK ratio, p-CREB/CREB ratio and c-fos level in the hypothalamus and hippocampus and this enhancement in morphine-treated animals, was more considerable than that in saline-treated animals.

Radon exhalation rate and natural radionuclide content in building materials of high background areas of Ramsar, Iran.

Radon exhalation rates from building materials used in high background radiation areas (HBRA) of Ramsar were measured using an active radon gas analyzer with an emanation container. Radon exhalation rates from these samples varied from below the lower detection limit up to 384 Bq.m(-2) h(-1). The (226)Ra, (232)Th and (40)K contents were also measured using a high resolution HPGe gamma- ray spectrometer system. The activity concentration of (226)Ra, (232)Th and (40)K content varied from below the minimum detection limit up to 86,400 Bq kg(-1), 187 Bq kg(-1) and 1350 Bq kg(-1), respectively. The linear correlation coefficient between radon exhalation rate and radium concentration was 0.90. The result of this survey shows that radon exhalation rate and radium content in some local stones used as basements are extremely high and these samples are main sources of indoor radon emanation as well as external gamma radiation from uranium series.

Estimates of the occupational exposure to tenorm in the phosphoric acid production plant in Iran.

Phosphate rock is used world wide for manufacturing phosphoric acid and several chemical fertilisers. It is known that the phosphate rock contains various concentrations of uranium, thorium, radium and their daughters. The subject of this study is the evaluation of the radiation exposure to workers in the phosphoric acid production plant due to technologically enhanced naturally occurring radioactive materials that can result from the presence of naturally occurring radioactive materials in phosphate ores used in the manufacturing of phosphoric acid. Radiation exposure due to direct gamma radiation, dust inhalation and radon gas has been investigated and external and internal doses of exposed workers have been calculated. Natural radioactivity due to (40)K, (226)Ra and (232)Th have been measured in phosphate rock, phosphogypsum, chemical fertilisers and other samples by gamma spectrometry system with a high-purity germanium. The average concentrations of (226)Ra and (40)K observed in the phosphate rock are 760 and 80 Bq kg(-1), respectively. Annual effective dose from external radiation had a mean value of ∼0.673 mSv y(-1). Dust sampling revealed greatest values in the storage area. The annual average effective dose from inhalation of long-lived airborne was 0.113 mSv y(-1). Radon gas concentrations in the processing plant and storage area were found to be of the same value as the background. In this study the estimated annual effective doses to workers were below 1 mSv y(-1).

Evaluation of occupational exposure to naturally occurring radioactive materials in the Iranian ceramics industry.

Zircon contains small amounts of uranium, thorium and radium in its crystalline structure. The ceramic industry is one of the major consumers of zirconium compounds that are used as an ingredient at ∼10-20 % by weight in glaze. In this study, seven different ceramic factories have been investigated regarding the presence of radioactive elements with focus on natural radioactivity. The overall objective of this investigation is to provide information regarding the radiation exposure to workers in the ceramic industry due to naturally occurring radioactive materials. This objective is met by collecting existing radiological data specific to glaze production and generating new data from sampling activities. The sampling effort involves the whole process of glaze production. External exposures are monitored using a portable gamma-ray spectrometer and environmental thermoluminescence dosimeters, by placing them for 6 months in some workplaces. Internal routes of exposure (mainly inhalation) are studied using air sampling, and gross alpha and beta counting. Measurement of radon gas and its progeny is performed by continuous radon gas monitors that use pulse ionisation chambers. Natural radioactivity due to the presence of ²³8U, ²³²Th and 4°K in zirconium compounds, glazes and other samples is measured by a gamma-ray spectrometry system with a high-purity germanium detector. The average concentrations of ²³8U and ²³²Th observed in the zirconium compounds are >3300 and >550 Bq kg⁻¹, respectively. The specific activities of other samples are much lower than in zirconium compounds. The annual effective dose from external radiation had a mean value of ∼0.13 mSv y⁻¹. Dust sampling revealed the greatest values in the process at the powdering site and hand weighing places. In these plants, the annual average effective dose from inhalation of long-lived airborne radionuclides was 0.226 mSv. ²²²Rn gas concentrations in the glaze production plant and storage warehouse were found to range from 10 to 213 Bq m⁻³. In this study, the estimated annual effective doses to exposed workers were <1 mSv y⁻¹.

Calcium and phosphorus metabolism disturbances after renal transplantation.

INTRODUCTION: Several studies have noted that, despite beneficial correction of abnormalities of mineral metabolism after successful renal transplantation, renal functional recovery is incomplete. Also, persistence of hyperparathyroidism and metabolic acidosis among patients with chronic impairment of graft function together with the use of loop diuretics and immunosuppressive drugs with adverse effects may alter mineral metabolism. We determined calcium and phosphorus levels in recipients. METHODS: This cross-sectional study enrolled 398 recipients in 2 medical centers in Iran from 1988 to 2004 to evaluate serum calcium and phosphorus levels after 1 month in relation to graft and patient survivals. Cyclosporine was the constant part of the immunosuppressive treatment in all study subjects. RESULTS: The median follow-up time was 8 months (range, 1-180 months). One and 10-year survival rates of patients were 97.9% and 91.1%. Mean (SD) serum calcium levels before and after transplantation were 8.79 (1.26) and 8.50 (1.39) mg/dL, respectively (P=.020). The mean (SD) phosphate levels before and after transplantation were 6.43 (2.42) and 3.64 (1.71) mg/dL, respectively (P=.000). There was no significant difference in survival considering changes in serum calcium and phosphorus levels. There was no correlation between serum calcium and phosphorus level changes among study patients. CONCLUSIONS: Despite reports suggesting hypercalcemia as a posttransplantation finding, we did not observe this condition, but, consistent with other reports in this field, we observed a significant decrease in serum phosphorus levels showing correction of this mineral level.

[Inhibition of the yeast respiratory system by Zn-protoporphyrin and effect of photolysis of this substance]. / Etude de l'inhibition du système respiratoire des levures par la Zn-protoporphyrine en fonction de la photolyse de cette substance.

We have shown earlier that yeast cells grown in synthetic mediums supplemented with Zn++ accumulate large amounts of Zn-protoporphyrin within their mitochondria. This accumulation is accompanied by an inhibition of respiration (3). This study deals with the effect of light on the respiratory inhibition and the release of respiratory control which are observed if Zn-protoporphyrin is added to isolated mitochondria which are initially devoid of this pigment. In addition, we have studied the effect of light on the respiratory inhibition exerted by Zn-protoporphyrin accumulated in vivo. The following results were obtained: 1) The light-induced destruction of Zn-protoporphrin which had been added in vitro to Zn-protoporphyrin-free mitochondria significantly inhibits respiration and phosphorylation. Under these conditions, the extent of the inhibitions increases with the concentration of the added Zn-protoporphyrin and the duration of illumination. 2) Accumulation of Zn-protoporphyrin within the cells causes an inhibition of the respiratory activities and the activities of succinate-cytochrome c reductase and NADH-cytochrome c reductase of the mitochondria. Illumination of the isolated mitochondria from Zn-protoporphyrin-containing cells enhances the inhibition of these activities. No light-induced inhibition of these activities is observed with mitochondria from cells devoid of Zn-protoporphyrin.