Live birth after in vitro maturation in women with gonadotropin resistance ovary syndrome: report of two cases.

PURPOSE: Gonadotropin-resistant ovary syndrome (GROS) is a rare endocrine disorder that causes hypergonadotropic hypogonadism, amenorrhea, and infertility. This study reports live birth in two women with GROS who underwent fertility treatment with in vitro maturation (IVM). METHODS: Both patients had primary infertility, amenorrhea (primary and secondary), typical secondary sexual characters, elevated gonadotropin levels, normal ovarian reserve, normal chromosomal characteristics, and previous nonresponsiveness gonadotropin stimulations. One patient had polymorphism of the follicle-stimulating hormone receptor, which is a predictor of poor ovarian response. Given unresponsiveness to exogenous gonadotropin stimulations, IVM with human chorionic gonadotropin priming (hCG-IVM) was performed in both patients. All transferrable embryos were vitrified. RESULTS: Both patients achieved pregnancy after their first frozen embryos transfer, and each delivered a healthy baby boy. CONCLUSIONS: These results suggest that IVM should be a first-line therapeutic option for patients with GROS.

Breast Milk for Preterm Multiples: More Proteins, Less Lactose.

Exclusive breastfeeding is currently recommended until at least 6 months of postnatal age, due to maternal breast milk (BM) unique composition and beneficial properties. In fact, BM modifies itself according to gestational age (GA) at birth, adapting its composition to neonatal requests during lactation. Multiple births represent about 3% of the whole pregnancies; such neonates result more vulnerable than full-term newborns, due to lower GA and birth weight (BW) and the higher incidence of perinatal complications. Although an adequate nutrition is fundamental for twins and other multiples, studies on this topic are lacking. We collected and analyzed BM from mothers of 19 twins and 5 triplets showing GA < 33 weeks and BW < 1500 g, comparing it to a control group of 28 preterm singletons. As a result, at GA ≤ 28 weeks, we observed that protein content is higher in BM for multiples (1.53 vs. 1.29 g per 100 ml), lactose concentration is greater in BM for singletons (6.72 vs. 6.34 g per 100 ml) and GA results the most relevant factor influencing BM protein composition. BM for multiples results higher in proteins and lower in lactose, if compared with singleton's samples; this could promote and sustain growth and organ development in this vulnerable category. BM from multiples shows a trophic and immunologic role, since these neonates often show lower GA and BW instead of singletons. These findings could help in optimizing nutritional strategies and improving BM individualized fortification.

Twin Conception in Sheep Leads to Impaired Insulin Sensitivity and Sexually Dimorphic Adipose Tissue and Skeletal Muscle Phenotypes in Adulthood.

Twins are often born small and early and have increased risk of obesity and diabetes later in life. Twin conception in sheep, regardless of whether the pregnancy continues as twins or is reduced to singleton in early gestation, alters offspring growth trajectory and body composition in young adulthood. We hypothesized that twin conception would result in insulin resistance in adulthood, with insulin-resistant adipose tissue and skeletal muscle phenotypes. At 3 years of age, body weight was not different among singletons, twins, and reductions; females weighed less than males. Singletons were leaner than reductions, with twins intermediate. Twins and reductions had decreased insulin sensitivity compared with singletons (singletons: mean [standard error of the mean]: 4.75 [0.4], twins: 3.34 [0.3], reductions: 3.67 [0.2] mg·I µU-1·kg-1·min-1, P < .01). There were no group differences in adipocyte size, adipose tissue, or circulating tumor necrosis factor α, monocyte chemoattractant protein 1, or interleukin 6 concentrations. In males, omental and subcutaneous adipose SLC2A4 was 1.5- to 2.0-fold greater in twins and reductions than in singletons ( P < .01) and SLC2A1 was greater in reductions than in singletons. Skeletal muscle IRS-1 was decreased in male twins but increased in female twins, compared to singletons ( P ≤ .01), with no effect on reductions in either sex. Skeletal muscle SLC2A4 was decreased in female twins and reductions but elevated in male twins and reductions compared to singletons ( P ≤ .01). We conclude that adult twin insulin resistance is not due to adipose tissue phenotype, but potentially phenotypic effects in skeletal muscle, and obesity is a result of twin conception per se with its origins in early gestation.

Gestational diabetes mellitus in women with multiple pregnancies: is the metabolic abnormality milder?

OBJECTIVE: We aimed to compare maternal characteristics and dysglycemia after delivery in women with gestational diabetes mellitus (GDM) according to pregnancy being multiple (MP) or singleton (SP). The hypothesis was that women with GDM and MP would have a milder glycemic abnormality before and after pregnancy than those with SP. METHODS: We performed a cohort study of 2908 women giving birth between 1986 and 2009. Logistic regression was performed to discriminate between MP and SP after anamnestic pre-pregnancy characteristics. Kaplan-Meier and Cox regression analyses were performed to assess if MP was independently associated with both impaired fasting glucose (IFG)/impaired glucose tolerance (IGT) and diabetes after delivery. RESULTS: Family history of diabetes was the only independent anamnestic pre-pregnancy characteristic discriminating MP versus SP, OR 2.04 (95% CI 1.12, 3.70, p 0.019). The median time to progress to IFG/IGT was 7.52 years in SP (95% CI 6.92, 8.13) and 7.41 in MP (95% CI 3.84, 10.98), ns and the progression to DM did not differ. In addition, MP was not associated to IFG/IGT or to DM in the Cox regression analysis. CONCLUSIONS: In this cohort of women with GDM, those with MP did not demonstrate a lesser degree of dysglycemia after controlling for other pregnancy characteristics and pregnancy-independent factors.

Nutrition for Multiples.

In 2012 there were 135,943 infants of multiple pregnancies born in the United States, nearly a 2-fold increase since 1980, with twins accounting for 96% of all multiple births. To date, most perinatal morbidities associated with multiple births have proven resistant to technological or pharmaceutical interventions. Maternal nutrition can have a profound effect on the course and outcome of multiple pregnancy, with the goal of achieving optimal intrauterine growth and birthweights, and minimizing prenatal and perinatal complications for the mother and her children.

Reproducción Asistida: ¿Debemos solo transferir un embrión? / Assisted Reproduction: Should be transferred only one embryo?

OBJETIVO: Comparar resultados obstétricos y perinatales de embarazos únicos espontáneos con los obtenidos por reproducción asistida, tanto únicos como gemelares. MATERIAL Y MÉTODO: Estudio descriptivo observacional retrospectivo. Se analizan un total de 1 096 embarazos de pacientes que acuden a control obstétrico en una clínica privada y se clasifican en tres grupos según el modo de concepción y número de fetos: Grupo 1, embarazo único espontáneo; Grupo 2, embarazo único por técnica de reproducción asistida (TRA) y, Grupo 3, embarazo gemelar por reproducción asistida. Se analizan los resultados obstétricos y perinatales entre los tres grupos. Se considera que hay diferencia estadística con un valor menor a <0.01 utilizando la prueba de chi cuadrado. RESULTADOS: La tasa de diabetes gestacional fue mayor en el grupo de embarazo único por TRA (23,78%) comparada a la de embarazo único espontáneo (6,66%) y gemelar por TRA (0%), presentando diferencia estadísticamente significativa. Hubo mayor presencia de anemia gestacional en el grupo de embarazo único espontáneo (76,66%) que en los grupos de TRA (19,51% y 31,25% para único y gemelar respectivamente) con diferencia estadística. Se presenta diferencia significativamente estadística a favor del grupo de embarazo gemelar por TRA en cuanto a un mayor número de casos de amenaza de parto pretérmino (25 %), una mayor ganancia de peso materno a final del embarazo (12,33 ± 3,2 Kg), un mayor número de nacimientos por cesárea (81,25%), menor edad en semanas de gestación al parto (35,28 ± 3,6), menor peso del recién nacido (2 388,69 ± 378 g) y un mayor número de casos que requieren ingreso a unidad de cuidados intensivos neonatales (18,75%). CONCLUSIONES: El embarazo gemelar tiene un peor pronóstico perinatal que el embarazo único sea espontáneo o por TRA

OBJETIVE: To determine the correlation between the semen parameters (concentration, motility, morphology and vitality) and sperm DNA integrity, using the test Halosperm. Material And METHODS: Prospective study conducted at the Laboratory of Andrology of Assisted Reproduction Laboratory FERTILAB, Lima - Peru, from August 2012 to March 2013. 282 patients were analyzed. We have established the DNA fragmentation index (IFA) from patients with semen sample normozoospermic and patients with semen sample altered in some parameters. We used two types of threshold (18% and 30%) and were correlated with semen parameters. In the total population was determined the Spearman correlation between sperm parameters and the IFA. RESULTS: It was determined that the value of the median the IFA of the patients with semen sample normozoospermic was significantly lower than that patients with semen sample altered in some parameters (12.8% vs 19.0, P=0.000). Using threshold of 18% was determined the there significant difference between the median of the two groups (Group 1: ≤ 18% y Group 2: > 18%) in age (37 years vs 40 years, P = .002), sperm concentration (82.30 X 106/ml vs 58.00 X 106/ml, P = 0.046), progressive motility (45.80% vs 27.40, P = 0.000), normal morphology (12.50% vs 9.00%, P = 0.000) and sperm vitality (89.0% vs 78.0%, P = 0.000). Using the threshold of 30 % was found significant difference between the median of the two groups (Group 1: ≤ 30% y Group 2:> 30%) in age (39 years vs 44 years, P=0.000), sperm concentration ( 78.00 X 106/ml vs. 36.75  X 106/ml, P = 0.015 ), motility progressive (40.85% vs 22.38 %, P=0.000 ), normal morphology (12.0 % vs 6.0 %, P=0.004) and sperm vitality (85.0 % vs 72.5%, P=0.000). Also identified an inverse correlation between the IFA and concentration (r=- 0.219 P=0.000), progressive motility (r=-0.452 P=0.000), normal morphology (r=-0.322 P=0.000) and vitality (r = -0.452 P = 0.000) in the total population. We determined a significant positive correlation between age and the IFA (r=0.267 P = 0.000). CONCLUSIONS: The results indicate that the level of DNA damage in sperm patients with semen sample altered in some parameters is significantly higher than in patients with semen sample normozoospermic. It has been shown that the semen parameters (concentration, motility, morphology and vitality) are highly negatively correlated with sperm DNA fragmentation index. It showed a positive correlation between age and the IFA

Embryo number and periconceptional undernutrition in the sheep have differential effects on adrenal epigenotype, growth, and development.

Exposure to poor maternal nutrition around the time of conception results in an early prepartum activation of the fetal pituitary-adrenal axis and in increased adrenal growth and stress response after birth associated with epigenetic changes in a differentially methylated region (DMR) of adrenal IGF2/H19. We have determined the effects of maternal undernutrition during the periconceptional period (PCUN: 70% of control intake from 60 days before until 6 days after conception) and early preimplantation period (PIUN: 70% of control intake for 6 days after conception) on fetal plasma ACTH and cortisol concentrations and fetal adrenal ACTHR, StAR, 3ßHSD, CYP11B, CYP17, TGFß1, IGF1, IGF1R, IGF2, and IGF2R mRNA expression and the methylation level of sites within the DMRs of IGF2/H19 and IGF2R in the adrenal of twin and singleton fetuses at 136-138 days gestation. Being a twin resulted in a delayed prepartum increase in fetal ACTH and in a lower cortisol response to CRH in the control but not PCUN and PIUN groups. PCUN, but not PIUN, resulted in an increase in adrenal weight and CYP17 expression in singletons, a decrease in adrenal IGF2 expression in singletons, and an increase in adrenal IGF2R expression in both twins and singletons. IGF2/H19 and IGF2R DMR methylation levels and ACTHR expression were lower in the twin adrenal. Thus, exposure of the oocyte and embryo to maternal undernutrition or to the environment of a twin pregnancy have differential effects on epigenetic and other factors that regulate fetal adrenal growth and IGF2 and IGF2R expression.

Stability of genomic imprinting and gestational-age dynamic methylation in complicated pregnancies conceived following assisted reproductive technologies.

For the past three decades, assisted reproductive technologies (ART) have revolutionized infertility treatments. The use of ART is thought to be safe. However, early investigations suggested that children born as a result of ART had higher risk of diseases with epigenetic etiologies, including imprinting disorders caused by a lack of maternal methylation at imprinting control elements. In addition, large epidemiology studies have highlighted an increased risk of obstetric complications, including severe intrauterine growth restriction (IUGR) in babies conceived using ART. It is plausible that the increased frequency of IUGR may be due to abnormal imprinting because these transcripts are key for normal fetal growth and development. To address this, we have collected a large cohort of placenta and cord blood samples from ART conceptions and compared the imprinting status with appropriate non-ART population. Using a custom DNA methylation array that simultaneously quantifies 25 imprinted differentially methylated regions, we observed similar epigenetic profiles between groups. A multiplex Sequenom iPLEX allelic expression assay revealed monoallelic expression for 11 imprinted transcripts in our placenta cohort. We also observe appropriate gestational age-dependent methylation dynamics at retrotransposable elements and promoters associated with growth genes in ART placental biopsies. This study confirms that children conceived by ART do not show variability in imprinted regulation and that loss-of-imprinting is not commonly associated with nonsyndromic IUGR or prematurity.

The hedgehog system in ovarian follicles of cattle selected for twin ovulations and births: evidence of a link between the IGF and hedgehog systems.

Hedgehog signaling is involved in regulation of ovarian function in Drosophila, but its role in regulating mammalian ovarian folliculogenesis is less clear. Therefore, gene expression of Indian hedgehog (IHH) and its type 1 receptor, patched 1 (PTCH1), were quantified in bovine granulosa (GC) or theca (TC) cells of small (1-5 mm) antral follicles by in situ hybridization and of larger (5-17 mm) antral follicles by real-time RT-PCR from ovaries of cyclic cows genetically selected (Twinner) or not selected (control) for twin ovulations. Expression of IHH mRNA was localized to GC and cumulus cells, whereas PTCH1 mRNA was greater in TC than in GC. Estrogen-active (E-A; follicular fluid concentration of estradiol > progesterone) versus estrogen-inactive follicles had a greater abundance of mRNA for IHH in GC and PTCH1 in TC. Abundance of IHH mRNA in GC was not affected by cow genotype, whereas TC PTCH1 mRNA was less in large E-A follicles of Twinners than in controls. In vitro, estradiol and wingless-type (WNT) 3A increased IHH mRNA in IGF1-treated GC. IGF1 and BMP4 treatments decreased PTCH1 mRNA in small TC. Estradiol and LH increased PTCH1 mRNA in IGF1-treated TC from large and small follicles, respectively. In summary, functional status of ovarian follicles was associated with differences in hedgehog signaling in GC and TC. We hypothesize that as follicles grow and develop, increased free IGF1 may suppress expression of IHH mRNA by GC and PTCH1 mRNA by TC, and these effects are regulated in a paracrine way by estradiol and other intra- and extragonadal factors.

Relationship between plasma progesterone concentration and number of conceptuses and their growth in superovulated cattle.

Elevated concentrations of circulating progesterone (P4) in the immediate post-ovulation period are associated with advancement of conceptus elongation in cattle. Superovulated (SOV) cattle have not only elevated plasma P4 concentrations but also multiple embryos in the uterus because of the formation of multiple corpora lutea. We examined the relationship between plasma P4 concentration and uterine glucose level in the immediate post-ovulation period and the presence and growth of multiple conceptuses in SOV cattle. SOV cattle were artificially inseminated with frozen-thawed semen at standing estrus (day 0), and the conceptuses were recovered by nonsurgical flushing of the uterus on day 13. In the SOV cattle, there were quadratic relationships between plasma P4 concentration on days 4, 5 and 7 and conceptus length and between number of conceptuses in the uterus and conceptus length. These results suggest that conceptus growth in SOV cattle is regulated by both systemic P4 level and number of conceptuses and that there are ranges of plasma P4 concentrations and numbers of conceptuses in the uterus that are suitable for conceptus growth and development. Plasma P4 concentrations on days 5 and 7, but not the numbers of conceptuses, were quadratically correlated with uterine glucose levels on day 13 in SOV cattle. In addition, conceptus length was positively correlated with uterine glucose level in SOV cattle. Accordingly, regardless of the number of conceptuses in the uterus, the plasma P4 concentration was well correlated with the regulation of conceptus growth via changes in uterine glucose levels in SOV cattle.

Effect of prolonged in vivo administration of progesterone in pregnancy on myometrial gene expression, peripheral blood leukocyte activation, and circulating steroid hormone levels.

OBJECTIVE: We aimed to investigate the effects of progesterone on gene expression and function of both myometrium and circulating leukocytes. METHODS: We recruited women participating in a randomized clinical trial of progesterone to prevent preterm delivery. These participants had a twin pregnancy and were managed in 1 of 2 tertiary referral centers. Participants were treated with progesterone (90 mg vaginally) or placebo from 24 to 34 weeks of pregnancy. The outcome measures were myometrial and leukocyte gene expression and expression of cell surface markers in circulating leukocytes, all quantified ex vivo. RESULTS: Prolonged in vivo administration of progesterone inhibited myometrial expression of connexins 26 and 43, endothelial nitric acid synthase (eNOS), and the prostaglandin receptor EP2 ex vivo. Administration of progesterone also increased numbers of circulating neutrophils while decreasing lymphocyte proportions and decreasing neutrophil CD11b expression. CONCLUSION: The observed effects of prolonged in vivo administration of progesterone will minimize the ability of the uterus to contract as a synctium and the ability of peripheral blood leukocytes to migrate into the myometrium during parturition. We suggest that these are putative mechanisms by which progesterone might prevent preterm birth in women at high risk.

Parto diferido del segundo gemelo / Delayed-interval delivery in twin pregnancy

La incidencia de embarazos múltiples se ha incrementado en los últimos años. Las pacientes con embarazos múltiples están en riesgo de parto prematuro con una alta asociación a mortalidad y morbididad perinatal. El parto del primer gemelo en una gestación múltiple va normalmente seguido por el parto del siguiente en un corto periodo. Es raro observar un intervalo prolongado entre el parto de los dos fetos de un embarazo múltiple. Nosotros reportamos 7 casos de embarazos múltiples con un parto diferido del segundo gemelo. Basándonos en nuestra experiencia y en la revisión de la literatura, concluimos que el parto diferido del segundo gemelo en edades gestacionales extremas, con un control exhaustivo de las condiciones fetales y maternas, está recomendado para mejorar la supervivencia y disminuir la morbilidad en el segundo gemelo (AU)

The incidence of multiple pregnancies has increased in the last few years. Patients with multiple pregnancies are at risk of preterm delivery associated with high perinatal mortality and morbidity. Delivery of the first twin in a multiple gestation is usually followed by delivery of the second twin shortly thereafter. A prolonged interval between delivery of the fetuses in a multiple pregnancy is infrequent. We report seven cases of multiple pregnancies with delayed- interval delivery of the second twin. On the basis of our experience and a review of the literature, we conclude that delayed delivery of the second twin in very preterm gestational ages, with careful observation of fetal and maternal status, is recommended to improve survival and decrease morbidity in the second twin (AU)

Diferencias en el metabolismo lipídico, niveles de homocisteína, folato y vitamina B12 entre gestaciones gemelares y únicas / Differences in lipid metabolism, homocysteine levels, serum folic acid and vitamin B12 between single and twin pregnancies

Antecedentes: Se están usando diferentes tipos de suplementación nutricional durante la gestación, sin embargo, existen escasez de estudios que se centren en los resultados metabólicos de los mismos. Objetivos: Comprobar si mediante la suplementación se logra el control del impacto de los múltiples cambios metabólicos que conlleva una gestación múltiple, usando análisis comunes. Método: Comparamos tres diferentes grupos de pacientes entre la semana gestacional 25 y 27. Un grupo de 46 gestaciones únicas, un grupo de 38 gestaciones gemelares espontáneas y otro grupo de 32 gestaciones gemelares tras técnicas de reproducción asistida. Comparamos niveles de colesterol LDL, HDL, triglicéridos, folato sérico, vitamina B12, homocisteína, hemoglobina, creatinina y plaquetas. Resultados: Los niveles de triglicéridos fueron más altos en ambos grupos de gemelares (p<0,001) y los niveles de colesterol fueron similares en los tres grupos. Las concentraciones de folato sérico fueron mayores en las gestaciones gemelares (p<0,001). El resto de parámetros estudiados fueron similares en los tres grupos. Conclusión: La suplementación siguiendo las recomendaciones de la ACOG y la SEGO es efectiva en el control de colesterol LDL, HDL, folato sérico, vitamina B12, homocisteína, hemoglobina, creatinina y plaquetas. Sólo los niveles de triglicéridos no se relacionan con la suplementación y pueden estar relacionados con mayor incidencia de colestasis y preeclampsia en gestaciones múltiples.

Background: Different ways of nutritional supplementation are being advised by physicians during pregnancy, but there is a lack of literature focused on its metabolical results. Aims: Check if the control of the impact that the multiple metabolic changes related to multiple pregnancies have on the mother's homeostasis is achieved with the supplementation, through common used blood tests. Method: We compare three different groups of patients between the 25 and 27th week of gestation. One group of 46 single pregnancies, a second group of 38 spontaneous twin pregnancies and another group of 32 twin pregnancies after assisted reproduction techniques. We compare the levels of LDL, HDL-cholesterol, triglycerides, serum folic acid, Vit B12, homocysteine, haemoglobine, creatinine and platelets. Results: The levels of triglycerides were higher in both groups of twin pregnancies (p<0.001), but the levels of cholesterol remain similar. Folic acid concentrations were higher in the groups of twin pregnancies (p<0.001). The rest of levels studied were similar in the 3 groups. Conclusion: The supplementation following the ACOG and SEGO recommendations seems to be effective in controlling the values of LDL, HDL-cholesterol, serum folic acid, Vit B12, homocysteine, haemoglobine, creatinine and platelets. Only the levels of triglycerides appear unaffected by the supplementation and can be related with the higher incidence of preeclampsia and cholestasis seemed among twin pregnancies.

Effects of twin pregnancy and dry period feeding strategy on milk production, energy balance, and metabolic profiles in dairy cows.

The present study evaluated the interaction of pregnancy type [PT; single (S) vs. twin (T)] and dry period feeding management [D; close-up (CU) diet (NE(l) = 1.54 Mcal/kg of DM)] throughout the entire dry period (8W) vs. far-off (FO) diet (NE(l) = 1.32 Mcal/kg of DM) from 60 to 21 d before expected calving date (ECD) followed by CU diet until calving (3W). Treatments were arranged in a 2 x 2 factorial with a randomized block design with primiparous (n = 8) and multiparous (n = 39) Holstein cows. We hypothesized that increasing the duration of feeding a CU diet would improve metabolic status and lactation performance for cows with T, but not for cows with S. All cows were fed similarly in late lactation (90 to 60 d before ECD; diet NE(l) = 1.58 Mcal/kg of DM) and in early lactation (calving to 105 DIM; diet NE(l) = 1.71 Mcal/kg of DM). Prepartum DMI as percentage of BW did not differ (P > 0.10) with D but tended to be greater (P = 0.10) for cows with S than with T. Cows with T tended to have greater (P = 0.08) BW than cows with S, but conceptus-free BW was less (P = 0.001) for cows with T than for cows with S. No differences (P > 0.10) were detected in prepartum BCS or BCS change with PT or D. Energy balance (EB) was greater for cows with S than with T (P < 0.001) and for cows fed 8W vs. 3W (P = 0.01). Cows with T had greater (P < 0.001) NEFA and a tendency for greater liver triglycerides (TG; P = 0.07) and plasma beta-hydroxybutyrate (BHBA; P = 0.06) than cows with S. Prepartum cows fed 3W had greater (P = 0.01) liver TG and greater (P = 0.02) plasma NEFA, but less (P = 0.02) plasma BHBA than cows fed 8W. Plasma glucose (P < 0.004) and liver glycogen (P = 0.02) were less for cows with T but were not affected (P > 0.10) by D. Postpartum, there was no effect (P > 0.1) of PT or D on mean DMI as percentage of BW, BW, and BCS, but there was an interaction (P = 0.02) of PT x D for mean BCS. Cows that calved T were in a more positive (P = 0.004) EB than cows that calved S. Milk production was 5.2 kg/d greater (P = 0.04) for cows fed 8W; however, they were in less (P = 0.01) EB than cows that received 3W. Postpartum cows that calved T had decreased concentrations of plasma NEFA (P = 0.02) and liver TG (P = 0.04) but greater concentrations of plasma glucose (P = 0.03) than cows that calved S. Plasma BHBA (P = 0.07) and NEFA tended (P = 0.06) to be greater for cows that received 8W than 3W. Neither PT nor D affected (P > 0.1) plasma glucose and liver glycogen. There was a tendency for an interaction of PT x D for plasma NEFA and liver TG. In contrast to our hypothesis, response to D was independent of PT. Based on milk production data from the present experiment, 8W is a more desirable feeding strategy than 3W.

Renal regulation of calcium-phosphate metabolism in single- and twin pregnant goats.

The aim of this study was the clearance assessment of renal function in single- and twin-pregnant goats and a comparison of selected parameters associated with calcium-phosphate management in terms of litter size. Clearance studies were carried out on 16 pregnant Polish White Improved goats (8 single pregnancies and 8 twin pregnancies). It was demonstrated that the kidneys of pregnant goats regulate the calcium-phosphate balance to a great extent. In spite of observed differences in parameters of renal functions, plasma calcium and inorganic phosphorus concentrations were comparable with normal reference values in further weeks of pregnancy. Significant differences in renal function between single and twin pregnancies were not observed. Gut absorption and/or bone tissue metabolism seem important for the maintenance of appropriate calcium-phosphate status.

Follicle and hormone dynamics in single versus double ovulating heifers.

Follicles > or = 5 mm were ablated at 4 day post-ovulation in heifers to induce a follicular wave, and prostaglandin F(2alpha) was given at day 6 to increase the incidence of double ovulations. Follicle diameters and plasma hormone concentrations were compared between single ovulators (n=12) and double ovulators (n=8). In double ovulators, the interval from follicle deviation to the peak of the pre-ovulatory LH surge was shorter (1.9+/-0.2 vs 2.5+/-0.2 days; P<0.02) and diameter of the largest pre-ovulatory follicle was smaller (12.2+/-0.5 vs 13.3+/-0.3 mm; P<0.02). The LH concentrations of the pre-ovulatory surge did not differ between single and double ovulators for 24 h on each side of the peak. When data were normalised to LH peak, the peak of the pre-ovulatory FSH and oestradiol (E(2)) surges occurred in synchrony with the peak of LH surge for both groups. Concentration of FSH for 24 h on each side of the peak showed a group effect (P<0.0001) from lower concentration in the double ovulators. A group-by-hour interaction (P<0.008) for E(2) reflected greater concentration in the double ovulators before and at the peak. Results indicated that two pre-ovulatory follicles resulted in an earlier and greater E(2) increase, leading to lower FSH concentration, an earlier LH surge, and ovulation at a smaller diameter. In conclusion, the difference in hormone concentrations during the pre-ovulatory period was an effect rather than a cause of double ovulations.

Inhibition of brain prostaglandin endoperoxide synthase-2 prevents the preparturient increase in fetal adrenocorticotropin secretion in the sheep fetus.

Maturation of the fetal hypothalamus-pituitary-adrenal axis is critical for the timely somatic development of the fetus and readiness for birth. Recently, we proposed that prostaglandin generation within the fetal central nervous system is critical for the modulation of hypotension-induced fetal ACTH secretion. The present study was designed to test the hypothesis that the preparturient increase in fetal ACTH secretion is dependent upon fetal central nervous system prostaglandin synthesis mediated by the activity of prostaglandin endoperoxide synthase (PGHS)-2 (cyclooxygenase-2) in the fetal brain. We performed two studies in chronically catheterized fetal sheep. In the first study, we infused nimesulide or vehicle intracerebroventricularly (i.c.v) into singleton fetal sheep and collected blood samples until spontaneous parturition. Nimesulide significantly delayed parturition, and inhibited fetal ACTH and proopiomelanocortin secretion but did not prevent the preparturient increase in fetal plasma cortisol concentration. In the second study, we used twin fetuses. One fetus received intracerebroventricular nimesulide and the other intracerebroventricular vehicle. Nimesulide reduced brain tissue concentrations of prostaglandin estradiol, while not affecting plasma prostaglandin E(2) concentrations, demonstrating an action restricted to the fetal brain. Nimesulide reduced PGHS-2 mRNA and increased PGHS-2 protein, while not altering PGHS-1 mRNA or protein in most brain regions, suggesting an effect of the inhibitor on PGHS-2 turnover and relative specificity for PGHS-2 in vivo. We conclude that the preparturient increase in fetal ACTH and proopiomelanocortin is dependent upon the activity of PGHS-2 in the fetal brain. However, we also conclude that the timing of parturition is not solely dependent upon ACTH in this species.

Placental vascularity and growth factor expression in singleton, twin, and triplet pregnancies in the sheep.

For singleton, twin, and triplet pregnancies, uteri were collected on day 140 of pregnancy. For each ewe (n = 18), placentomes were fixed by arterial perfusion supplying the fetal (cotyledon) and maternal placenta (caruncle). Tissue sections were stained for determination of vascularity by image analysis. Further, protein expression for factor VIII, vascular endothelial growth factor (VEGF) and its receptor, VEGFR1, as well as basic fibroblast growth factor (FGF2) and its receptor, FGFR, in tissue sections was determined by immunohistochemistry and image analyses. Cotyledonary and caruncular samples were analyzed for expression of mRNA for Vegf and its two receptors, Vegfr1 and Vegfr2, as well as Fgf2 and Fgfr. Fetal number did not affect placental capillary density or factor VIII expression, whereas increased fetal number reduced total cotyledon and caruncle capillary volume. While expression of Vegf, Vegfr1, Vegfr2, and Fgfr mRNA in cotyledonary but not caruncular tissue was greater in twin pregnancies compared to singleton and triplet pregnancies, protein expression of VEGF in the placentome decreased with increasing numbers of fetuses, VEGFR1 did not change, and FGFR was greater in twin versus singleton and triplet pregnancies. Fetal number did not affect the expression of Fgf2 mRNA in placental tissues, whereas FGF2 protein expression was less in triplet compared to singleton and twin pregnancies. Reduced fetal and placental weights in twins and/or triplet pregnancies are associated with an overall decrease in total placental vascularity, VEGF and FGF2 and/or FGFR protein expression, but not in angiogenic factor mRNA expression or VEGFR1 protein expression in sheep.

Chronic stretching of amniotic epithelial cells increases pre-B cell colony-enhancing factor (PBEF/visfatin) expression and protects them from apoptosis.

In normal pregnancy, the fetal membranes become increasingly distended towards term and in multifetal gestations they become over-distended. Apoptosis of the amniotic epithelium increases with advancing gestation and may contribute to fetal membrane weakening and rupture. The effects of chronic static stretching for 36h have been investigated using primary amniotic epithelial cells. Pre-B cell colony-enhancing factor (PBEF) is a stretch-responsive cytokine and expression of its gene, intracellular and secreted protein were all significantly increased by 4h and its secretion sustained over 36h, contrasting with the rapid increase and decline in expression of IL-8. Increased expression of SIRT1 and decreased p53 paralleled the changes in PBEF, are known to be responsive to PBEF, and contribute to cell survival. Distension had no effects on proliferation or necrosis but protected the cells from apoptosis, knocking-down PBEF with antisense probes abrogated this protective effect. There was increased immunostaining of PBEF in the compact layer of the amnion in multifetal tissues and significantly fewer apoptotic amniotic epithelial cells. These results show that chronic stretching of the amniotic epithelial cells increases PBEF expression, which protects them from apoptosis.

Placental expression of soluble fms-like tyrosine kinase 1 is increased in singletons and twin pregnancies with intrauterine growth restriction.

OBJECTIVE: Intrauterine growth restriction (IUGR) is characterized by decreased placental perfusion. Low oxygen has been shown to increase soluble fms-like tyrosine kinase 1 (sFlt-1) expression in the human placenta. The objective of this study was to examine sFlt-1 expression in different types of IUGR pregnancies, including early-onset severe cases characterized by abnormal umbilical and uterine artery Doppler and discordant IUGR twins in which the normal cotwin represents the optimal control because both placentas share the same uterine environment. PATIENTS: Placentas from four subgroups were collected: early severe IUGR with umbilical artery absent end diastolic flow (n = 19), small for gestational age with normal uterine and umbilical artery Doppler (n = 11), severely growth-restricted dichorionic and monochorionic twins with abnormal umbilical artery Doppler (n = 9), preeclamptic twins (n = 3), and age-matched normal singletons (n = 19) and twin controls (n = 8). RESULTS: Expression of sFlt-1 mRNA and protein was significantly increased in IUGR placentas compared with small for gestational age and normal control placentas. sFlt-1 expression levels were also significantly greater in the small IUGR twin placentas from discordant twin pregnancies compared with the normal cotwin. In preeclamptic twins, sFlt-1 expression was increased in only one of the two placentas. CONCLUSIONS: Our results demonstrate that sFlt-1 expression is increased in severe IUGR placentas with abnormal umbilical artery Doppler of singletons and also in discordant IUGR twins. Reduced placental perfusion may contribute to the increased expression of sFlt-1 in IUGR pregnancies. Our data are compatible with differential sFlt-1 expression in placentas from discordant twins.