Celery oil modulates DEHP-induced reproductive toxicity in male rats

The objective of the study was to investigate the protective effect of Apium graveolens (AP) against di-(2-ethylhexyl) phthalate (DEHP)-induced testes injury in rats. Adult rats were divided into nine groups: (1) control group (no treatment); (2) corn oil (60 μg/kg body weight – bwt); (3) AP (50 μg/kg bwt); (4) 300 mg DEHP/kg bwt; (5) 500 mg DEHP/kg bwt; (6) 1000 mg DEHP/kg bwt; (7) 300 mg DEHP/kg bwt + AP; (8) 500 mg DEHP/kg bwt + AP; and (9) 1000 mg DEHP/kg bwt + AP. Oral administration of treatments was performed daily for 6 weeks. DEHP decreased (p < 0.01) body weight, testis weight and serum concentrations of testosterone, cholesterol and total proteins. Moreover, DEHP increased (p < 0.001) total antioxidant capacity in the testis and plasma DEHP level. In addition, DEHP decreased mRNA expression of two testicular steroidogenic enzymes: 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase. DEHP also caused atrophy, vacuolar degeneration and aspermia of the seminiferous tubules. AP administered concurrently with DEHP effectively alleviated most of the DEHP-induced effects. In conclusion, in male rats, DEHP had adverse effects on the testis including inhibition of androgen production. A concurrent administration of A. graveolens (celery oil) protected the testis against DEHP-induced toxicity.     

Administration of recombinant bovine somatotropin (rbST) at the time of breeding in superovulated fertile and subfertile ewes

Two experiments were conducted to evaluate the effects of the administration of 125 mg rbST at the time of breeding on fertile and subfertile superovulated ewes – in terms of the ovulation rate, embryo yield and embryonic development. In addition, the effects of the treatment on the plasma progesterone concentrations and interferon-tau (IFNT) expression in the embryos were evaluated in the fertile animals. In the first experiment, carried out on fertile superovulated ewes, the treatment significantly increased the ovulation rate (p < 0.05) and shifted the embryo distribution towards more advanced stages of development (p < 0.01). Plasma progesterone concentrations increased faster in the treated ewes, being significantly different (p < 0.05) between groups at day 6 post-breeding. In the second experiment, embryonic development was more advanced in fertile, but not in subfertile superovulated ewes. The expression of IFNT was significantly (p < 0.01) decreased in blastocyst embryos obtained from rbST treated animals, compared with non-treated ewes. It is concluded that the administration of rbST to subfertile ewes at the time of breeding does not stimulate embryonic development as it does when administered to fertile animals.     

Expression of orexin receptors 1 (OX1R) and 2 (OX2R) in the porcine pituitary during the oestrous cycle

Orexin A and B, also termed hypocretin 1 and 2, are associated with the stimulation of food intake and arousal. The biological actions of the hormones are mediated via two distinct G protein-coupled receptors, termed orexin receptor 1 (OX1R) and orexin receptor 2 (OX2R). OX1R is selective for orexin A and OX2R binds orexin A and orexin B with similar affinity. The present study analyzed mRNA and protein expressions of OX1R and OX2R in adenohypophysis (AP) and neurohypophysis (NP) of cycling pigs. The tissue samples were harvested on days 2–3, 10–12, 14–16, and 17–19 of the oestrous cycle. Using quantitative real-time PCR higher OX1R gene expression was detected in AP on days 2–3 relative to days 10–12, 14–16 and 17–19 (p < 0.05). In NP the OX1R mRNA level was elevated on days 10–12 compared to the remaining stages (p < 0.05). OX2R gene expression in AP was the lowest on days 10–12 (p < 0.05 compared to days 2–3 and 17–19) and the expression peak occurred on days 17–19 (p < 0.05 vs. the all studied stages). In NP the highest (p < 0.05) expression of OX2R mRNA was noted on days 17–19 in relation to the remaining periods. OX1R protein content in AP was greatest on days 10–12 (p < 0.05), whereas in NP it was greatest on days 2–3 and 14–16 (p < 0.05 vs. days 10–12 and 17–19). In both cases the lowest OX1R protein expression was observed during follicular phase (p < 0.05 in relation to three remaining studied stages). OX2R protein in AP was lower (p < 0.05) on days 2–3 and 14–16 compared to days 10–12 and 17–19. In NP the lowest (p < 0.05) expression of this protein was on days 17–19 and the highest on days 10–12 (p < 0.05 compared to days 2–3 and 17–19). In summary, the present findings provide the first evidence that OX1R and OX2R mRNAs and proteins occur in the pituitary of the pig and indicate the dependence of orexin receptor expression on the endocrine reproductive state.     

Effect of dl-malate on mixed ruminal microorganism fermentation using the rumen simulation technique (RUSITEC)

The objective of this study was to investigate the effects of dl-malate on the in vitro fermentation of a 50 : 50 forage : concentrate diet using the rumen simulation technique (RUSITEC) and to compare these effects with those induced by the addition of propionate. The RUSITEC system consisted of eight vessels: three of them received daily 5.62 mmol of dl-malate, three vessels received daily 5.62 mmol of propionate and two vessels received no additives (control). After an adaptation period of 11 days the main fermentation parameters were determined for five consecutive days. There were no significant differences between treatments either in pH or in the daily production of NH₃–N. Compared to control diet, the addition of dl-malate resulted in an increase (p < 0.05) of hemicellulose disappearance and a trend (p < 0.10) towards a greater disappearance of dry matter, organic matter and neutral detergent fibre. Differences between dl-malate and propionate in diet degradation were not significant. dl-malate treatment resulted in an increase (p < 0.01) of daily propionate production, and a decrease (p < 0.05) in the amount of both methane (mmol/g DM fermented) and l-lactate (mg/day). Compared to propionate, dl-malate produced an increase (p < 0.05) of acetate production and tended to produce a lower amount of propionate (p < 0.10). In conclusion, these results seem to indicate that dl-malate enhanced the in vitro fermentation by increasing production of propionate and digestibility of hemicellulose.     

Fluctuations in energy-related metabolites during the peri-parturition period in Lori-Bakhtiari ewes

This study describes the fluctuations in serum energy-related metabolites during a period of 2 weeks before, to 2 weeks after parturition in Lori-Bakhtiari ewes. The effect of parity was also studied. Blood profiles were determined in 60 healthy pregnant ewes with single (n = 30) and twin (n = 30) lambings. Blood was collected from each ewe on days 14 and 7 prepartum, and days 7 and 14 postpartum to determine the serum non-esterified fatty acid (NEFA), β-hydroxybutyrate (BHBA), glucose, cholesterol, blood urea nitrogen (BUN) and calcium (Ca) levels. The age of the ewes had no significant effect on the energy metabolism indicators. Serum NEFA, BHBA, glucose, BUN and calcium concentrations recorded peak levels 7 days before parturition. However, NEFA and BHBA recorded significant changes (P < 0.05) during the peri-parturition period. All metabolites changed significantly in ewes carrying twin-bearing ewes, compared to single-bearing ewes. Serum BHBA concentrations recorded positive correlations with the serum NEFA (P < 0.01) and cholesterol (P < 0.05), while blood glucose had negative correlations (P < 0.01) with NEFA, BHBA and Ca. Blood NEFA and BHBA recorded positive correlations (P < 0.05) with the BUN levels and negative correlations (P < 0.05) with Ca. The results showed that blood NEFA and BHBA levels are sensitive indicators of the energy balance during the peri-parturition period in ewes.     

The effect of the absence or presence of a corpus luteum on the ovarian follicular population and serum oestradiol concentrations during the estrous cycle in Sanjabi ewes

The aim of this study was to determine if the presence or absence of a corpus luteum (CL) during estrous synchronization in ewes can affect the ovarian follicular population and the serum oestradiol concentrations. The estrous cycles of 197 Sanjabi ewes were synchronized using a 12-day treatment with intravaginal progestagen sponges (Chronogest^®). Estrus was detected in 144 ewes, 27–39 h after sponge removal. Blood samples were taken daily from day 2 and continued for 19 days and analyzed for serum oestradiol concentration. Nine ewes were slaughtered on each experimental day (days 1–16 after estrus) for ovary collection. The ovaries per ewe were classified as those without, or with one or two CL's, for each slaughter day. Visible follicles on the surface of the ovaries were classified, based on their diameter, into (i) very small (<2 mm), (ii) small (2–3.4 mm), (iii) medium (3.5–5 mm) and (iv) large (>5 mm) categories, and the respective numbers recorded. Results indicated, the number of ovarian follicles to decrease (P < 0.01) from days 1 to 5 of the cycle and showed a significant increase on day 7. Numbers were high again on day 11 and decreased (P < 0.01) on day 16 of the estrous cycle. The serum oestradiol concentrations were significantly higher (P < 0.001) in the double than in the single ovulating animals (one or two CL's, respectively) on days 2–0. However serum levels were also significantly higher (P < 0.001) in single, than twin ovulating animals on days 4–5 and 12–16 of the estrous cycle. There were no significant differences in the total number of very small follicles between animals without and those with two CL's. The number of small, medium and large follicles in ewes, with or without a CL on the ovary was significantly higher (P < 0.01) than ewes with two ovulations at certain stages of the estrous cycle. The present study provides evidence of differences in the follicular ovarian population in ovaries without CL's and double ovulations. The existence of an intraovarian effect of the CL numbers on follicular population is demonstrated.     

The effect of machine or hand milking on milk production,composition and SCC in mountainous Greek breed (Boutsiko) ewes

The aim of this work was to determine the effect of two different milking methods (machine versus hand milking) on milk yield, milk composition and somatic cell counts (SCC) in mountainous Greek Boutsiko ewes. Ninety-six ewes were divided in two groups of 48 ewes each. Milk yield, lactation number (parity) and type of lambing were taken into consideration for the separation of the ewes. One group was milked by machine and the other by hand. Milk production, milk composition (with Milkoscan 104) and SCC (with Fossomatic) were performed every 14 days, in the morning and evening milking; during the experimental period seven samplings were performed in total. The results of this study have shown that ewes milked by machine gave a greater daily milk yield as compared to those milked by hand (698 ml versus 652 ml; P < 0,05) and less log SCC (5.37 versus 5.62; P < 0,05). The composition of milk did not differ between the two treatments. Also, no phenotypic correlations were established between SCC and milk production or composition, apart from a negative correlation, which was noted between SCC and the percentage of lactose. The results of this experiment have shown that ewes of the Boutsiko breed can be milked by machine without any problems.     

The induction of oestrus in ewes during the non-breeding season using pre-used CIDRs and oestradiol-17β treatment

The fertility obtained in sheep after the use of intravaginal progesterone devices is related to the content of progesterone of the device. The hypothesis of this study was that the reproductive response of anoestrous ewes to the ram-effect could be improved by the administration of oestradiol-17β in conjunction with CIDRs treatment—using previously used CIDRs in a 5-day progestagen priming. Therefore, the objective was to determine if oestradiol-17β treatment increases fertility of anoestrous ewes primed with used CIDRs and stimulated by the ram-effect. The hypothesis was tested with CIDRs that had been previously used for 12 or 18 days. The trial was performed during the non-breeding season using 158 Corriedale ewes. Ewes had been isolated from rams since Day −35 (Day 0 = introduction of the rams). A CIDR (0.3 g progesterone, InterAg, Hamilton, New Zealand) was inserted on Day −5 in all ewes with CIDR that had been previously used for 12 days (n = 62) or 18 days (n = 96). Also on Day −5, 29 and 53 ewes that had received CIDRs of 12 or 18 days, respectively, received an intra-muscular treatment of 50 μg of oestradiol-17β (E groups). The ewes that did not receive the oestradiol-17β treatment remained as the control group (C group). Overall the treatment groups were thus: C12 (n = 33), C18 (n = 43), E12 (n = 29), and E18 (n = 53). On Day 0 all CIDRs were withdrawn, and ewes were placed with 18 rams and 20 ewes hormonally induced to exhibit oestrus. Sexual receptivity of ewes treated with CIDRs was estimated from marks on the rumps of the ewes daily from Day 0 to Day 5, and the pregnancy status diagnosed with transrectal ultrasonography on Day 40. The percentage of ewes exhibiting oestrus and pregnancy rates were lower in ewes synchronized with previously used CIDRs for 18 days, compared to those used for 12 days. The responses of ewes in oestrus were 39.4, 14.0, 65.5, and 32.1% for the C12, C18, E12, and E18 groups respectively, with pregnancy rates of 30.3, 14.0, 34.5, and 17.0%. Administration of oestradiol-17β increased the frequency of oestrous response in ewes that were treated with CIDRs previously used for 12 days (P < 0.05), but not in those treated with CIDRs used for 18 days. It could be concluded that the administration of oestradiol-17β only improved the percentage of ewes responding to oestrus when CIDRs previously used for 12 days were used for 5 days before the introduction of rams. No positive effect on fertility was observed irrespective of the period during which CIDR had been previously used.     

Effects of uterine and lactational exposure to di-(2-ethylhexyl) phthalate on spatial memory and NMDA receptor of hippocampus in mice

Di-(2-ethylhexyl) phthalate (DEHP) is an environmental endocrine disrupter. Currently, little is known about neurodevelopmental toxicity of DEHP in wildlife and humans. The present study investigated the effects of DEHP, focusing on the changes in the behavior of offspring mice at the ages of 6 and 12 w, respectively, following utero and lactational exposure to DEHP (10, 50, and 200 mg/kg/d) from gestation day 7 through postnatal day 21. The results of open field tasks showed that DEHP increased the grooming of males at age 6 w and females at age 12 w but decreased the frequency of rearing of 6-w-old females and the number of grid crossings of 12-w-old females. In the Morris water maze task, 50 and 200 mg/kg/d DEHP significantly prolonged the time of searching the hidden platform in water maze and reduced the time staying in the target quadrant during a probe trial of 6-w-old male mice, but not of 6-w-old females nor 12-w-old mice of both sexes, suggesting an impaired spatial learning and memory among younger males after perinatal exposure to DEHP. Western blot analyses further showed that DEHP at 50 and 200 mg/kg/d decreased the levels of the N-methyl-d-aspartic acid (NMDA) receptor subunits NR1 and NR2B in the hippocampus of 6-w-old males. These results suggest that uterine and lactational exposure to low doses of DEHP sex-specifically impacted behaviors, including locomotion activity and spatial memory, via the concomitant inhibition of the NMDA receptor of the hippocampus in offspring mice.     

Correlations between ovarian follicular blood flow and superovulatory responses in ewes

The primary goal of this study was to employ ultrasonography to examine the ovaries of ewes undergoing superovulatory treatment for correlations between antral follicular blood flow and ovarian responses/embryo yields. Five Santa Inês ewes were subjected to a short- (Days 0–6, Group 1) and five to a long-term progesterone-based protocol (Days 0–12, Group 2) to synchronize estrus and ovulations after the superovulatory treatment. Porcine FSH (pFSH, 200 mg) was administered in 8 decreasing doses over 4 days, starting on Days 4 and 10 in Groups 1 and 2, respectively. After CIDR removal, all ewes were bred by a ram and embryos were recovered surgically 7 days later. Transrectal ovarian ultrasonography was performed the day before and on all 4 days of the superovulatory treatment. Both an arbitrary-scale [(0) non-detectable; (1) small; (2) moderate; (3) intense blood flow] and quantitative analysis of the blood flow area were used to assess the follicular blood flow in color Doppler images. There were no significant correlations between the arbitrary blood flow scores and superovulatory responses in the ewes of the present study. However, there was a positive correlation between the quantitative estimates of follicular blood flow on the final day of the superovulatory treatment, and the number (DA: r = 0.68, P < 0.05; DA/TA × 100%: r = 0.85, P < 0.05) and percentage (DA: r = 0.65, P < 0.05; DA/TA × 100%: r = 0.91, P < 0.001) of unfertilized eggs (DA: Doppler area, TA: total area of the largest ovarian cross section). This experiment presents a commercially practical tool for predicting superovulatory outcomes in ewes and evidence for the existence of follicular blood flow threshold that may impinge negatively on oocyte quality when surpassed during hormonal ovarian superstimulation.     

Supplemental energy and protein increase use of sagebrush by sheep

The ability of sheep to consume big sagebrush (Artemisia tridentata Nutt.), a shrub that contains terpenes, may be improved with supplemental energy and protein needed for detoxification processes. We compared feeding behavior of supplemented and unsupplemented ewes grazing separately in three 0.5 ha plots of 40 m × 120 m during October 2001. Supplemented ewes were offered a choice of a high-energy (50% corn and 50% beet pulp—3.62 Mcal/kg DE and 9.5% CP) and a high-protein (85% alfalfa and 15% soybean meal—2.89 Mcal/kg DE and 15% CP) supplement for 15 min/day. Supplemented ewes consumed 492 g (S.E. = 24.3) of the high-energy supplement and 140 g (S.E. = 9.8) of the high-protein supplement. The incidence of feeding on sagebrush was recorded using scan sampling at 10-min intervals from 09:00 to 17:00 h every other day for 10 days. Incidence of feeding on sagebrush increased gradually for both treatments from 0.4% on day 1 to 28% on day 10 (P < 0.01), but supplemented ewes increased feeding frequency on sagebrush to nearly twice that of unsupplemented ewes by the end of the trial. Averaged throughout the trial, incidence of eating sagebrush was twice as high for supplemented compared to unsupplemented ewes (22% versus 11% of the total time feeding, P < 0.01). The results of this study show that supplemental macronutrients increase use of sagebrush by ewes. Thus, strategic supplementation during fall and winter may be a way to increase intake of sagebrush by sheep and enhance and maintain biodiversity in areas of the sagebrush steppe where sagebrush is deemed too abundant.     

The effect of GnRH,eCG and progestin type on estrous synchronization following laparoscopic AI in ewes

The aim of the experiment was to evaluate the effects of GnRH and/or eCG and progestin type (implant versus CIDR) on the induction of estrus and pregnancy rate following laparoscopic AI (LAI) with frozen semen. In the first trial, ewes (n = 129) were treated with norgestomet implants for 14 days. At implant removal ewes received eCG (400 IU) and/or GnRH (25 μg) 36 h after removal, resulting in control, eCG, GnRH, and eCG/GnRH groups (n = 30–34/group). In trial 2, ewes (n = 36) were treated with intravaginal fluorogestone acetate sponges (FGA) or CIDR for 12 days. After withdrawal, half of the ewes from each progestin group received eCG (400 IU), resulting in sponge, sponge/eCG, CIDR and CIDR/eCG groups (n = 8–10/group). In both trials, estrous activity was assessed using a vasectomized ram from the time of progestin removal to laparoscopic AI with frozen semen 58–60 h (trial 1) or 54–56 h (trial 2) following cessation of treatment. In trial 1, GnRH decreased (P < 0.05) the percentage of ewes in estrus (GnRH, 75.8% versus control, 93.8% versus eCG/GnRH, 94.1%), however pregnancy rates were similar in all groups (control, 53.1%; eCG, 70.0%; GnRH, 51.5%; eCG/GnRH, 55.9%, respectively). In trial 2, neither the type of progestin nor eCG treatment effected the percentage of ewes in estrus (sponge, 75.0%; sponge/eCG, 100.0%; CIDR, 100.0%; CIDR/eCG, 90.0%). However, pregnancy rates following LAI were higher (P < 0.05) when ewes were treated with eCG (progestin + eCG, 73.7% versus progestin alone, 41.2%). Results demonstrate that the source of progestin does not influence the expression of estrus or the proportion of ewes pregnant following LAI. When progestin treatment protocols are used in combination with eCG, pregnancy rates can be increased. A dose of GnRH near the end of progestin treatment may decrease the estrous response, by inducing ovulation before normal expression of estrus.     

Reproductive performance and milk yield in Awassi ewes following crossbreeding

The objective of this study was to evaluate the effect of crossbreeding Awassi ewes with either Charollais or Romanov sires, on pre-weaning lamb production as reflected by reproductive performance and milk production in the ewes. Two hundred and five, 3- to 6-year-old multiparous ewes of three breeds were allocated to three groups [Awassi (A; n = 56), F₁ Romanov × Awassi (RA; n = 78) and F₁ Charollais × Awassi (CA; n = 71)]. Upon lambing, ewes and their offspring were placed in a large pen in which they remained until the end of the trial. Body weight (BW) and body condition score (BCS) of the ewes and the BW of the lambs were recorded weekly from birth to weaning at 70 days of age. Milk production was recorded weekly from parturition to weaning. Pregnancy rates were not influenced by breed-type. Multiple births and the fecundity of ewes were greater (P < 0.05) in the RA group, compared to the A and CA groups—while the weaning percentage was similar between the breeds. Lamb birth and weaning weights were similar, while the kg lamb born per kg ewe lambed was greater (P < 0.05) in the RA group, compared to the CA group. The CA ewes recorded a higher BW (P < 0.01) and BCS (P < 0.01) than the other breed groups. Awassi ewes produced more milk (P < 0.01) throughout the observation period. Milk ash percentage was higher (P < 0.05) in the CA group, while the percentage of crude protein and dry matter was similar for the different groups. Results of the present study indicate that crossbreeding of Awassi ewes with Charollais and Romanov breeds decreased milk production in the progeny, without affecting lamb growth. The crossbreeding of Awassi with Charollais resulted in improved BW and BCS, while the crossbreeding of Awassi ewes with Romanov resulted in improved reproductive performance of the crossbreds. Crossbreeding Awassi ewes with exotic rams can thus be conducted to increase the number of lambs produced per ewe.     

Feeding,resting and social behaviour in ewes housed in two different group sizes

The aim of this experiment was to investigate the effects of increased group size on eating- and resting behaviour, aggression and feed intake in housed ewes. During an initial period of 14 days 36 adult (2–6 years old) ewes of the domestic Norwegian Dala breed were divided into four groups of 9. In the second period (14 days), these ewes were merged into one group of 36 ewes. This experiment was repeated with a second batch of ewes, but this time starting with a group of 36 individuals in the first period, then splitting them up into four groups of 9 ewes in the second period. From 24 h video recordings we scored activity behaviours using instantaneous sampling every 10 min. Aggressive interactions were continuously observed the first 10 min every hour during the 24 h (4 h in total). A mixed statistical procedure with group size, day, batch and the interactions between them were included as fixed effects, whereas individual and group were specified as random effects.Ewes in large groups (36) had a larger variation in lying time at day one (P < 0.01), less synchronized lying (P < 0.05) and eating behaviour (P < 0.01), and spent less time queuing at the feed barrier (P < 0.001) compared to in the small group size (9). There were no effects of group size on aggressive interactions or feed intake.In conclusion, a larger group size decreased synchrony in resting and feeding behaviour and reduced the time spent queuing in front of the feed barrier. It is possible that the aggression level in sheep is more sensitive to changes in space allowance than to changes in group size per se.     

The effects of post-stroke upper-limb training with an electromyography (EMG)-driven hand robot

Loss of hand function and finger dexterity are main disabilities in the upper limb after stroke. An electromyography (EMG)-driven hand robot had been developed for post-stroke rehabilitation training. The effectiveness of the hand robot assisted whole upper limb training was investigated on persons with chronic stroke (n = 10) in this work. All subjects attended a 20-session training (3–5 times/week) by using the hand robot to practice object grasp/release and arm transportation tasks. Significant motor improvements were observed in the Fugl-Meyer hand/wrist and shoulder/elbow scores (p < 0.05), and also in the Action Research Arm Test and Wolf Motor Function Test (p < 0.05). Significant reduction in spasticity of the fingers as was measured by the Modified Ashworth Score (p < 0.05). The training improved the muscle co-ordination between the antagonist muscle pair (flexor digitorum (FD) and extensor digitorum (ED)), associated with a significant reduction in the ED EMG level (p < 0.05) and a significant decrease of ED and FD co-contraction during the training (p < 0.05); the excessive muscle activities in the biceps brachii were also reduced significantly after the training (p < 0.05).     

The ghrelin activating enzyme ghrelin-O-acyltransferase (GOAT) is present in human plasma and expressed dependent on body mass index

Ghrelin is the only known peripherally produced and centrally acting peptide hormone stimulating food intake. The acylation of ghrelin is essential for binding to its receptor. Recently, the ghrelin activating enzyme ghrelin-O-acyltransferase (GOAT) was identified in mice, rats and humans. In addition to gastric mucosal expression, GOAT was also detected in the circulation of rodents and its expression was dependent on metabolic status. We investigated whether GOAT is also present in human plasma and whether expression levels are affected under different conditions of body weight. Normal weight, anorexic and obese subjects with body mass index (BMI) 30–40, 40–50 and >50 were recruited (n = 9/group). In overnight fasted subjects GOAT protein expression was assessed by Western blot and ghrelin measured by ELISA. GOAT protein was detectable in human plasma. Anorexic patients showed reduced GOAT protein levels (−42%, p < 0.01) whereas obese patients with BMI > 50 had increased concentrations (+34%) compared to normal weight controls. Ghrelin levels were higher in anorexic patients compared to all other groups (+62–78%, p < 0.001). Plasma GOAT protein expression showed a positive correlation with BMI (r = 0.71, p < 0.001) and a negative correlation with ghrelin (r = −0.60, p < 0.001). Summarized, GOAT is also present in human plasma and GOAT protein levels depend on the metabolic environment with decreased levels in anorexic and increased levels in morbidly obese patients. These data may indicate that GOAT counteracts the adaptive changes of ghrelin observed under these conditions and ultimately contributes to the development or maintenance of anorexia and obesity as it is the only enzyme acylating ghrelin.     

Effects of estradiol and progesterone on circulating LH and FSH secretion,and ovarian antral follicle growth in anestrous ewes

In the ewe, ovarian antral follicles emerge or grow in a wave-like pattern and each wave is preceded by a peak in the serum FSH level. The purpose of the current study was to investigate whether in anestrous Western White Face ewes, a combination of progesterone and estradiol affects the circulating FSH peak secretion and the number of small ovarian follicles. Five ewes were treated with subcutaneous silastic rubber implants (10 cm × 0.47 cm), containing 10% estradiol-17β w/w (controls) and 5 ewes were treated with the same estradiol implant, along with subcutaneous implants (11 cm × 0.48 cm) containing 10% progesterone w/w for 12 days. Daily transrectal ovarian ultrasonography and blood sampling was performed from 5 days before, to 9 days after the period of implantation. Blood samples were also taken every 12 min for a 6 h period on day −2, 6 and 13 prior to or after implant insertion (day 0, day of implant insertion). Pulsatility in the serum LH levels was eliminated by the implants (P < 0.05). During the implantation period, the serum FSH peak amplitude was lower in ewes treated with implants releasing estradiol and progesterone, compared to ewes treated with implants releasing only estradiol (P < 0.05). No follicular waves emerged during implant treatment in both groups (P < 0.05) and the number of serum FSH peaks did not differ during implantation, compared to before implantation. During the implantation period, the number of small follicles did not differ in ewes with implants releasing estradiol and progesterone, compared to ewes treated with implants releasing only estradiol. To conclude, supra-physiological concentrations of estradiol completely eliminated the serum LH pulsatality and suppressed the follicular wave emergence, while the FSH secretory peaks that preceded the follicular waves were not affected. Supra-physiological concentrations of estradiol-17β with physiological concentrations of progesterone decreased the serum FSH peak amplitude, eliminated the serum LH pulses, but did not decrease the size of the small follicle pool in anestrous ewes.     

Serum sialic acid changes in non-insulin-dependant diabetes mellitus (NIDDM) patients following bitter melon (Momordica charantia) and rosiglitazone (Avandia) treatment

Diabetes mellitus is associated with an increase in sialic acid concentration along with other complications. Sialic acid changes in NIDDM patients were investigated following bitter melon (55 ml/24 h) and rosiglitazone (4 mg/24 h) treatment. A total of 25 patients of both sexes were used in each experimental group. Patients following bitter melon treatment showed no significant difference of serum sialic acid (57.95±4.90 vs. 57.6±5.56 mg/dl, p=0.17) and serum glucose concentration (93.7±9.63 vs. 88.35±6.31 mg/dl, p=0.78) as compared to control subjects. However, the concentration of total cholesterol was significantly high in these patients as compared to control subjects (192±14.23 vs. 170.6±15.1 mg/dl, p<0.03) but within normal range (160–200 mg/dl), suggesting the significant hypoglycemic and lipid-lowering properties of bitter melon. The patients following rosiglitazone treatment showed a significant increase of serum sialic acid concentration (60.2±5.80 vs. 57.6±5.56 mg/dl, p=0.01) along with glucose (112±6.2 vs. 88.35±6.31 mg/dl, p<0.04) and total cholesterol concentration (216.45±20.2 vs. 170.6±15.1 mg/dl, p<0.01) as compared to control subjects. In addition six of the patients had retinopathy, two of whom were suffering also from myocardial infarction and they still had a higher serum sialic acid (61.05±1.20 mg/dl), glucose (187±2.11 mg/dl), total cholesterol (239.10±5.04 mg/dl) and triglyceride (183±4.14 mg/dl) concentration, indicating a poor response of these patients to rosiglitazone. Comparison of serum sialic acid concentration of patients, following bitter melon and rosiglitazone treatment revealed no significant difference but the study showed that bitter melon could be more effective in the management of diabetes and its related complications as compared to rosiglitazone.     

Th1/Th2/Th17/Treg cytokine imbalance in systemic lupus erythematosus (SLE) patients: Correlation with disease activity

AimImbalance of T-helper-cell (TH) subsets (TH1/TH2/TH17) and regulatory T-cells (Tregs) is suggested to contribute to the pathogenesis of Systemic lupus erythematosus (SLE). Therefore, we evaluated their cytokine secretion profile in SLE patients and their possible association with disease activity.MethodsSixty SLE patients, 24 rheumatoid arthritis (RA) patients and 24 healthy volunteers were included in this study. Demographic, clinical, disease activity and serological data were prospectively assessed. Plasma cytokines levels of TH1 (IL-12, IFN-γ), TH2 (IL-4, IL-6, IL-10), TH17 (IL-17, IL-23) and Treg (IL-10 and TGF-β) were measured by enzyme linked immunosorbent assays (ELISA).ResultsSLE patients were found to have significantly higher levels of IL-17 (p < 0.001), IL-6 (p < 0.01), IL-12 (p < 0.001) and IL-10 (p < 0.05) but comparable levels of IL-23 and IL-4 and slight reduction (but statistically insignificant) of TGF-β levels compared to controls. IL-6, IL-10 and IL-17 were significantly increased (p < 0.05) with disease activity. The RA group exhibited significantly higher levels of plasma IL-4 (p < 0.01), IL-6 (p < 0.05), IL-17 (p < 0.001), IL-23 (p < 0.01) and TGF-β (p < 0.5) and lower IFN-γ (p < 0.001) and IL-10 (p < 0.01) than those of healthy subjects.ConclusionOur study showed a distinct profile of cytokine imbalance in SLE patients. Reduction in IFN-γ (TH1) and TGF-β1 (Treg) with the elevation in IL-6 and IL-17 (TH17) could imply skewing of T-cells toward TH17 cells. Breaking TH17/Treg balance in peripheral blood may play an important role in the development of SLE and could be responsible for an increased pro-inflammatory response especially in the active form of the disease.     

Determination of trace endocrine disruptors in ultrapure water for laboratory use by the yeast estrogen screen (YES) and chemical analysis (GC/MS)

High purity water for endocrine disruptors (EDs) analysis in experimental tests is an indispensable requirement for the preparation of reagents and solutions employed in biological laboratories. Commercial ultrapure water may contain traces of organic compounds, which can interfere with in vitro bioassays carried out to detect the potential estrogen-like activity of pure compounds and complex mixtures. This paper shows that solid-phase extracts of different types of ultrapure water (UPW) purchased or produced in situ for laboratory analysis (mQ-UPW) may contain organic molecules able to antagonize the binding of E₂ to the human estrogen receptor α in the yeast estrogen screen (YES) assay. GC/MS analysis detected the presence of bis(2-ethylhexyl) phthalate (DEHP) (0.033 ppm ± 0.006) in mQ-UPW extracts. The dose–response curve of DEHP in the YES assay showed a relevant antagonist effect of this phthalate. Agreement between content of DEHP chemically detected in UPW extract and the magnitude of biological effects induced was pointed out. It would be appropriate that chemical analyses were complemented by biological tests to establish concentration limits for chemical contaminants in UPW that do not induce biological effects detectable in vitro. The yeast assay used in this study has previously proved to be a sensitive tool in assessing the presence of agonistic/antagonistic chemicals at the ng/l level in complex mixtures and may be successfully used to identify trace amounts of estrogenic/antiestrogenic chemicals, which can represent critical issues influencing the experimental results in environmental testing laboratories.