Serum concentrations of fibroblast growth factor 21 are elevated in patients with congenital or acquired lipodystrophy

ObjectivePatients with lipodystrophy (LD) suffer from loss of subcutaneous adipose tissue accompanied by dysregulation of several adipocyte-secreted factors. However, regulation of adipocyte-expressed fibroblast growth factor (FGF) 21 which acts in an insulin-mimetic, lipid-lowering, and anti-atherogenic manner has not been investigated in non-human immunodeficiency virus (HIV) LD.Material and methodsCirculating serum FGF21 levels were quantified in 37 patients with non-HIV LD and 37 controls matched for age, gender, and body mass index. Moreover, FGF21 plasma levels and mRNA expression were measured in LD mice and control animals. Additionally, serum FGF21 levels were assessed in 10 LD patients before and during metreleptin therapy.ResultsMedian FGF21 serum concentrations were significantly higher in LD patients (381.2 ng/l) as compared to the control group (231.2 ng/l; p = 0.023). There was an independent and positive association between circulating FGF21 and serum triglycerides (TG), as well as fibrate treatment, in multiple linear regression analysis. LD mice showed significantly upregulated FGF21 plasma levels (4.5-fold), as well as mRNA expression in various adipose tissue depots and liver as compared to controls (p < 0.05). Metreleptin treatment did not significantly alter circulating FGF21 levels in human subjects.ConclusionsSerum concentrations of FGF21 are elevated in patients with non-HIV LD with adipose tissue and liver being potential sources of increased production. TG and fibrate treatment are independent positive predictors of circulating FGF21.     

Differential activity of matrix metalloproteinases (MMPs) during photoperiod induced uterine regression and recrudescence in Siberian hamsters (Phodopus sungorus)

Siberian hamsters adapt to seasonal changes by reducing their reproductive function during short days (SD). SD exposure reduces uterine mass and reproductive capacity, but underlying cellular mechanisms remain unknown. Because matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are important in uterine development, parturition, and postpartum remodeling, their expression in uterine tissue from Siberian hamsters undergoing photoperiod-mediated reproductive regression and recrudescence was investigated. Female hamsters were exposed to long day (LD, 16L:8D, controls) or SD (8L:16D) for 3-12 weeks (regression); a second group was exposed to SD or LD for 14 weeks and then transferred to LD for 0-8 weeks (recrudescence). Hamsters were euthanized, uteri collected, and homogenates analyzed by gelatin zymography or Western blotting for MMP and TIMP protein levels. Uterine weight decreased (67-75%) at SD weeks 12-14 and increased post-LD transfer (PT) reaching LD values by PT week 2. MMP-2, but not MMP-9 activity was reduced by SD week 12 or 14 but increased to LD levels at PT week 2. MMP-3 expression increased at SD week 9 compared to other SD and LD groups. MMP-14 and -13 protein levels decreased at SD week 3 but returned to LD levels by SD week 6. During recrudescence, MMP-3 (PT weeks 0-2), MMP-13 (PT week 4), and MMP-14 (PT weeks 2, 4) protein levels were higher than LD. TIMP-1 and 2 were present at low levels. Significant and differential variations in uterine MMP activity/expression during photoperiod-induced regression and recrudescence were observed. These changes likely reflect increases in tissue remodeling during both the adaptation to SD and the restoration of reproductive function.     

Postpartum ovarian activity and uterine involution in non-seasonal Shiba goats,with or without nursing

Postpartum ovarian activity, uterine involution, and the relationship with hormonal profiles were studied in non-seasonal Shiba goats, with or without nursing of their kids. After parturition, does were allocated to one of two groups, with either weaning on the day of parturition (n = 3; non-nursing group) or weaning at 7–10 weeks after parturition (n = 4; nursing group). Blood sampling (starting 7 days prior to expected day of parturition) and transrectal ultrasound evaluations (starting 2 days after parturition) were conducted every other day or daily to monitor the follicular dynamics, uterus size, and the levels of plasma progesterone, blood glucose, and insulin concentration until at 3 weeks following the first postpartum ovulation. In the nursing group, blood samples were also collected every 10 min for an 8 h period on the day before weaning, 2 and 6 days after weaning, for the analysis of the pulsatile patterns of LH release. In all animals, the blood glucose concentrations increased transiently on the day before parturition and were significantly (p < 0.05) higher than those on the day of parturition (mean of 144 ± 48.8 mg/dl vs 63 ± 11.2 mg/dl). In the non-nursing group, the first postpartum ovulation was observed 9.3 ± 3.2 days after parturition, while in the nursing group, no ovulation occurred before weaning in any of the goats. Here ovulation was observed 18.8 ± 5.0 days after weaning, which was significantly (p < 0.05) later than in the non-nursing group. After first ovulation, all animals in both groups showed early luteal regression. No significant difference in the time required for the completion of uterine involution was recorded between the non-nursing and nursing groups (18.3 ± 4.2 days vs 19.3 ± 3.6 days, respectively), nor in the plasma LH pulse frequency obtained before (1.3 ± 0.5 pulses/8 h), 2 (1.3 ± 1.0 pulses/8 h) and 6 days (2.0 ± 0.8 pulses/8 h) after parturition, in the nursing group. It can be concluded that no ovulation occurs during nursing postpartum, suggesting that nursing is a determinant of the resumption of ovulation in these non-seasonal goats. Ultrasonic observations of the ovary suggest that uterine involution was not influenced by nursing or suckling.     

The proliferative phase underpins endometrial development: Altered cytokine profiles in uterine lavage fluid of women with idiopathic infertility

Endometrial gland development occurs during the proliferative phase of a woman’s menstrual cycle, laying the foundation for the subsequent receptive, secretory phase when pregnancy is established. Idiopathic infertility has been rarely investigated with respect to the proliferative phase endometrium. We investigated whether gland development and/or altered secretion of cytokines during the proliferative phase is associated with infertility. Area of the glandular epithelium (GE) was measured in proliferative phase endometrial tissue collected from fertile (n = 18) and infertile (n = 14) women. Cytokines were measured in proliferative phase uterine lavage of fertile (n = 15) and infertile (n = 15) women. Immunohistochemistry determined cellular localisation of transforming growth factor alpha (TGFα) and interferon gamma (IFNγ) in proliferative phase endometrial tissue. For statistical analysis the cohort was divided into women <35 years and ⩾35 years. There were no significant differences in GE area of infertile and fertile women. C-C motif chemokine 11 (P = 0.048), TGFα (P = 0.049), IFNγ (P = 0.033) and interleukin-1 alpha (P = 0.047) were significantly elevated in uterine lavage from infertile women <35 years compared to fertile but not in women ⩾35 years. TGFα and IFNγ localised predominantly to GE in both the fertile and infertile endometrium. The potential impact of this altered proliferative phase environment on subsequent receptivity is discussed.     

Valoración de niveles de testosterona mediante diferentes métodos analíticos en varones sanos

Plasma testosterone concentrations are essential for the diagnosis of several causes of hypogonadism, including late-onset hypogonadism. Defining the normal range for testosterone concentrations poses certain difficulties due to the changes that occur with age and the variability of the different analytical methods used.ObjectivesTo study normal ranges of testosterone in healthy young men and to compare the results of distinct analytical methods.Material and methodsWe recruited 20 healthy men with a mean age of 24.5 years (standard deviation (SD): 5.04) and a mean body mass index (BMI) of 23.8% (SD: 3.3). Total testosterone (TT) was measured by immunochemiluminescence (ICLA) and free testosterone (FT) by radioimmunoassay (RIA). Calculated free testosterone (FTc) and bioavailable testosterone (BT) were calculated using Vermeulen's formula. Serum lutropin (LH), follitropin (FSH) and sex hormone binding globulin (SHBG) were measured by immunoradiometric assays (IRMA).ResultsThe mean concentrations were 20 nmol/l (SD: 4.96) for TT, 0.054 nmol/L (SD: 0.01) for FT, 0.3834 nmol/L (SD: 0.09) for FTc and 9.9 nmol/L (SD: 2.8) for BT. There was no correlation between testosterone measured by different methods other than an association between FT and FTc (r=0.662, p<0.003) and between FTc and BT (r=0.979, p<0.0001). An inverse correlation was found between BMI and TT concentrations (r: ?0.52, p<0.017).ConclusionsThe normal range for testosterone in healthy young men should be established in each laboratory based on the analytical method used.     

Graded response to short photoperiod during development and early adulthood in Siberian hamsters and the effects on reproduction as females age

Short day (SD) lengths delay puberty, suppress ovulation, inhibit sexual behavior, and decelerate reproductive aging in female Siberian hamsters (Phodopus sungorus). To date, the modulation of the age-associated decline in reproductive outcomes has only been demonstrated in female hamsters experiencing different day lengths during development. To determine if developmental delay is necessary for photo-inhibition to decelerate reproductive aging, hamsters raised in LD were transferred to SD as young adults and remained there for 6 months. Females that demonstrated the most immediate and sustained photo-inhibition were found to have greater numbers of ovarian primordial follicles at advanced ages (9 and 12 months) than did females held in LD, nonresponders to SD, and females with a marginal SD-response. Similarly, for females raised in SD from conception to 6 months of age, prolonged developmental delay was associated with greater numbers of primordial follicles at later ages as compared to hamsters that became refractory to SD. A robust response to SD in juvenile and adult hamsters is associated with decelerated reproductive aging, which may result in greater reproductive success in older females as compared to age-matched individuals demonstrating a more modest response to SD.     

Effects of praeruptorin C on blood pressure and expression of phospholamban in spontaneously hypertensive rats

BackgroundThe traditional Chinese medicine Praeruptorin c (Pra-c) has many physiological and pharmacological effects, including antagonistic effects on blood pressure and calcium levels, maintenance of cellular calcium homeostasis, and improved cardiac systolic and diastolic function. It is potentially a novel and versatile drug for the treatment and prevention of cardiovascular diseases.ObjectiveTo explore the possible impact of Pra-c on blood pressure in SHR and its mechanism of action.Materials and methodsTwenty SHR were randomly divided into a Pra-c group [Pra-c was administered intragastrically, 20 mg kg⁻¹ d⁻¹, n = 10] or an untreated control group (n = 10), containing 10 age-matched SD rats. Each group of rats was followed for 8 weeks. Before and during the treatment, tail artery systolic blood pressure was measured using a tail-cuff every 2 weeks. After 8 weeks, the rats were sacrificed and RNA was extracted from homogenates of cardiac tissue. Tissue from the left ventricle was fixed, sectioned and H&E stained to assess possible changes in myocardial cell structure and morphology. Semi-quantitative RT-PCR was used to assess changes in phospholamban gene expression in treated and untreated rats.ResultsSHR treated with Pra-c for 8 weeks had a lower systolic pressure than untreated SHR (p < 0.05), two measures of cardiac damage, the heart mass index and left ventricle mass index (HMI and LVMI, respectively) were improved, and the level of PLB mRNA expression was lower in the untreated SHR group (p < 0.05).Discussion and conclusionWith continuous hypertension, SHR gradually formed or developed cardiac hypertrophy and fibrosis. Pra-c had a clear effect on blood pressure in SHR, and reversed SHR ventricular remodeling by upregulating the gene expression of sarcoplasmic reticulum PLB.     

Effect of arousing stimuli on circulating corticosterone and the circadian rhythms of luteinizing hormone (LH) surges and locomotor activity in estradiol-treated ovariectomized (ovx + EB) Syrian hamsters

In most proestrous hamsters, novel wheel exposure phase advances activity rhythms and blocks the preovulatory LH surge, which occurs 2 h earlier the next day. Because wheel immobilization does not prevent these effects we hypothesized that arousal alone blocks and phase advances the LH surge. Ovariectomized (ovx) hamsters received a jugular vein cannula and estradiol benzoate (EB) or vehicle was injected sc. The next day (Day 1), at zeitgeber time (ZT) 4–5 (ZT 12 = lights off), after obtaining a blood sample, each hamster was exposed to constant darkness (DD), and either remained in her home cage or was transferred to a new cage and exposed to a running wheel or a 2-hour arousal paradigm. Blood samples were obtained in dim red light and activity was recorded hourly until ~ ZT 10–11 on Days 1 and 2. For the next 1–2 weeks, activity was monitored in DD. Plasma LH and corticosterone were assessed by RIA. Novel wheel exposure or arousal at ZT 4 greatly attenuated the Day 1 LH surge in ovx + EB hamsters, and phase advanced the Day 2 LH surge by about 2 h. In proestrous hamsters, novel wheel exposure led to a prolonged (> 2 h) increase in corticosterone levels only when LH surges were blocked. Phase advances in activity rhythms were enhanced by estradiol and arousal. The results suggest that estradiol modulates the effectiveness of non-photic stimuli. The role of the increased activity of the hypothalamic–pituitary–adrenal axis associated with novel wheel-induced attenuation of LH surges in ovx + EB hamsters remains to be determined.     

C-type natriuretic peptide plasma levels are reduced in obese adolescents

The high prevalence of obesity in children may increase the magnitude of lifetime risk of cardiovascular disease (CD). At present, explicit data for recommending biomarkers as routine pre-clinical markers of CD in children are lacking. C-type natriuretic peptide (CNP) is assuming increasing importance in CD; in adults with heart failure, its plasma levels are related to clinical and functional disease severity. We have previously reported five different reference intervals for blood CNP as a function of age in healthy children; however, data on plasma CNP levels in obese children are still lacking. Aim of this study was to assess CNP levels in obese adolescents and verify whether they differ from healthy subjects. Plasma CNP was measured in 29 obese adolescents (age: 11.8 ± 0.4 years; BMI: 29.8 ± 0.82) by radioimmunoassay and compared with the reference values of healthy subjects. BNP was also measured. Both plasma CNP and BNP levels were significantly lower in the obese adolescents compared to the appropriate reference values (CNP: 3.4 ± 0.2 vs 13.6 ± 2.3 pg/ml, p < 0.0001; BNP: 18.8 ± 2.6 vs 36.9 ± 5.5 pg/ml, p = 0.003). There was no significant difference between CNP values in males and females. As reported in adults, we observed lower plasma CNP and BNP levels in obese children, suggesting a defective natriuretic peptide system in these patients. An altered regulation of production, clearance and function of natriuretic peptides, already operating in obese adolescents, may possibly contribute to the future development of CD. Thus, the availability of drugs promoting the action of natriuretic peptides may represent an attractive therapeutic option to prevent CD.     

Stem-swelling and photosynthate partitioning in stem mustard are regulated by photoperiod and plant hormones

The experiment was conducted to study the relationship between stem-swelling and photoperiod and growth hormones by comparing stem swelling with non-stem-swelling stem mustard (Brassica juncea var. tsatsai) plants about their growth characteristics and levels of endogenous gibberellin and cytokinin under different photoperiods. The results here showed that plant biomass was higher in 12-h photoperiod compared to that in long day (LD) and short day (SD), whereas stem growth was much stronger in LD compared to 12-h photoperiod and SD. Exogenous application of 1.0 mM gibberellic acid (GA₃) accelerated stem elongation in SD, but 8.9 μM benzyladenine (BA) failed. The shape of the swollen stem was also found to be associated with day length: a LD promoted stem elongation, while a 12-h photoperiod made the stem oval swollen. Also, stem was shown to have no sign of swelling in plants in SD with a relatively poor growth. The further studies showed that the largest proportion of ¹⁴C photosynthate was allocated to the swelling stems in stem-swelling plants, but to expanded leaves in non-stem-swelling plants, and endogenous gibberellin A₁ (GA₁) and zeatin + zeatin riboside (ZRs) were higher in LD compared to 12-h photoperiod and SD. These results from this experiment indicate that stem growth and swelling is a physiological process of hormonal control, and the photoperiod possibly exerts its influence by altering the balance between the levels of endogenous gibberellins and cytokinins.     

Limits to sustainable energy budget during lactation in the striped hamster (Cricetulus barabensis) raising litters of different size

A female animal appears to approach an upper limit to the rate of sustained energy intake/metabolic rate (SusEI/MR) during lactation. However, different species of animals may respond differently to the sustainable limit. Here, we measured energy budget during lactation in female striped hamsters raising litters of natural size (Con), and females whose litter size was manipulated during early lactation to support fewer or more pups (minus pups, MP or plus pups, PP). The striped hamsters significantly decreased their body mass and increased food intake from early to late lactation; and MP females had lower weight loss and food intake than the control and PP females. Litter size of the PP group decreased significantly over the period of lactation, and pups were weaned at a similar weight to that of the controls. MP females supported a significantly lower litter mass throughout lactation compared with the control and PP females, but during late lactation the pups from the MP group were significantly heavier. Resting metabolic rate (RMR) did not differ significantly between the three groups and the gross energy intake during peak lactation was 5.0×, 4.2× and 5.0 × RMR for the control, MP and PP females, respectively. Female striped hamsters reached a plateau in food intake at around 14 g/d during peak lactation, which might signify a limit of SusEI at 5.0 × RMR. However, it was not possible to determine whether the limitation on SusEI was imposed centrally by the capacity of the gastrointestinal tract to process food, peripherally by the capacity of the mammary gland to produce milk, or by the capacity of animals to dissipate heat.     

Age-specific breast,uterine and ovarian cancer mortality trends in Spain: Changes from 1980 to 2006

Introduction: Cancers of the breast, uterus and ovary are responsible for 30% of the cancer deaths in Spanish women. In recent decades, Spain has experienced important socioeconomic transformations, which may have affected mortality trends. We present the current situation of mortality in Spain due to cancers of the breast, uterus and ovary, as well as trends over 1980–2006. Methods: Data on population and deaths due to cancers of the breast, uterus and ovary were obtained from records of the National Statistics Institute. Overall and age-specific changes in mortality of these tumors were studied using change-point Poisson regression models. Results: Breast cancer was responsible for more than 140,000 deaths of females in 1980–2006. Trend analysis of breast cancer mortality of women of all ages showed that rates increased 2.9% annually until 1992 (95% confidence interval (CI) = 2.5, 3.3). After 1992, mortality declined steadily at a rate of ?2.1% per year (95% CI = ?2.4, ?1.8). The number of deaths due to cancers of the uterus was 49,287 between the years 1980 and 2006. Uterine cancer mortality registered a steady decrease of ?1.9% every year since 1980 (95% CI = ?2.1, ?1.8). Ovarian cancer caused 36,157 deaths during the same period, with rates in women older than 50 years more than ten-fold those of younger women. Trend analysis showed a sharp increase of mortality up to 1998 (4.4% annually; 95% CI = 3.9, 4.8) followed by a stabilization. Conclusion: The downturn observed in mortality for these tumors mainly reflects improved survival as a result of earlier diagnosis and better cancer treatments. Cancer management is moving in the right direction in Spain.     

Effects of total flavonoids of raspberry on perimenopausal model in mice

To study the effect of raspberry total flavonoids on perimenopausal model in mice. Blank group, sham operation, and the rest of the mice made the menopausal model. Choose 72 mice castrated completely random divided into 6 groups for the experiment, respectively: model group, gengnianan (GNA) capsule group, soybean isoflavone soft (SIS) capsule group, high, mid and low dose group of total flavonoids of raspberry (TFR). Animals in each group were given the corresponding drugs tenth days after operation, and were given intragastrical administration of once a day for continuous administration of 21 days. Each group of mice in the administration of 18 days to determine the number of autonomic activities within 5 min, in the administration of 19–20 days to determine the incubation of the mice first entry into the darkroom and the number of shocks into the darkroom within 5 min. At 2 h after the last administration (fasting for 12 h), mice were sacrificed and serum was collected. Serum levels of E2, T, LH and FSH were measured. Dissect the uterus, uterus, thymus and spleen. Weigh the wet weight and calculate the organ index, the morphological changes of uterus, thymus and spleen were observed. The results showed that the TFR had a good therapeutic effect on the perimenopausal model of mice after giving a high, mid and low dose of raspberry flavonoids for some time.     

Ultrasonographic imaging of the reproductive tract and surgical recovery of oocytes in Cebus apella (capuchin monkeys)

Two-dimensional real-time and Doppler ultrasonography are valuable non-invasive methods to assess reproductive anatomy and physiology. In adult, postpubertal female Cebus apella (capuchin monkeys), the objectives were to determine (1) uterine and ovarian dimensions, ovarian follicular dynamics, day of ovulation, and arterial blood flow of uterus and utero-ovarian ligament during the follicular phase of the menstrual cycle and (2) the number of oocytes aspirated from antral follicles at laparotomy. Based on two-dimensional, transabdominal B-mode ultrasonography, mean (± S.E.M.) length, height, width, and volume of the uterus were 17.9 ± 0.4, 12.4 ± 0.3, 13.6 ± 0.3 mm, and 1.55 ± 0.08 mL, respectively, and of the ovary were 13.4 ± 0.2, 8.2 ± 0.1, 7.7 ± 0.1 mm, and 4.5 ± 0.2 mL. Ovarian follicles were monitored for 6 days before ovulation, which occurred on day 9.3 ± 0.5 (range, days 7–11; day 1 = start of menses), with 10 of 12 ovulations in the right ovary. Diameter and volume of the preovulatory follicle were 10.1 ± 0.2 mm and 0.55 ± 0.03 mL (on the estimated day of ovulation) and of the CL were 8.1 ± 0.4 mm and 0.3 ± 0.05 mL. Resistivity and pulsatility indices were 0.86 ± 0.02 and 2.15 ± 0.11 for uterine arteries, and were 0.69 ± 0.04 and 1.63 ± 0.15 for the utero-ovarian ligament (UOL) artery; just prior to ovulation, both indices peaked (P < 0.05) in the uterine artery ipsilateral to the side of ovulation, but both reached a nadir (P < 0.05) in the UOL artery. In the absence of ovarian stimulation, 31 oocytes (diameter, 137 ± 10 μm) were aspirated (average of 2 oocytes/(female attempt)) on days 5, 7, and 9. In conclusion, transabdominal ultrasonography facilitated assessment of reproductive anatomy and physiology in C. apella adult females. Resistance and pulsatility indices of uterine and UOL arteries changed near the time of ovulation. Dominant follicles were easiest to aspirate at 8–9 mm in diameter (∼day 9), with intact cumulus-oocyte complexes recovered from ovarian follicles 2–9 mm in diameter.     

Serum levels of GDF15 are reduced in preeclampsia and the reduction is more profound in late-onset than early-onset cases

BackgroundPreeclampsia is a pregnancy specific disorder affecting 3–5% of pregnancies worldwide. It is clinically divided into early-onset and late-onset subtypes. Placental factors are involved in the pathogenesis of preeclampsia. Growth differentiation factor 15 (GDF15), a protein of the transforming growth factor beta superfamily, is highly expressed in the placenta. However, it is unclear whether the circulating levels of GDF15 are altered in preeclampsia at the time of or prior to disease presentation.MethodsSerum samples across three trimesters from 29 healthy pregnancies, third trimester sera from 34 women presenting with preeclampsia (early-onset n = 16, late-onset n = 18) and 66 gestation-age-matched controls, and sera at 11–13 weeks of pregnancy from women who later did (n = 36) or did not (n = 33) develop late-onset preeclampsia, were examined for GDF15 by ELISA.ResultsSerum GDF15 levels increased significantly with gestation in normal pregnancy. Serum GDF15 was significantly reduced in the third trimester in women presenting with preeclampsia compared to their gestation-age-matched controls. This reduction was apparent in both early-onset and late-onset subtypes, but it was more profound in late-onset cases. At 11–13 weeks of gestation, however, serum levels of GDF15 were similar between women who subsequently did and did not develop late-onset preeclampsia.ConclusionSerum GDF15 increased with gestation age, reaching the highest level in the third trimester. Serum GDF15 was significantly reduced in the third trimester in women presenting with preeclampsia, especially in late-onset cases. However, serum GDF15 was not altered in the first trimester in women destined to develop late-onset preeclampsia.     

PYY(3-36) into the arcuate nucleus inhibits food deprivation-induced increases in food hoarding and intake

Central administration of neuropeptide Y (NPY) increases food intake in laboratory rats and mice, as well as food foraging and hoarding in Siberian hamsters. The NPY-Y1 and Y5 receptors (Rs) within the hypothalamus appear sufficient to account for these increases in ingestive behaviors. Stimulation of NPY-Y2Rs in the Arcuate nucleus (Arc) has an anorexigenic effect as shown by central or peripheral administration of its natural ligand peptide YY (3-36) and pharmacological NPY-Y2R antagonism by BIIE0246 increases food intake. Both effects on food intake by NPY-Y2R agonism and antagonism are relatively short-lived lasting ∼4 h. The role of NPY-Y2Rs in appetitive ingestive behaviors (food foraging/hoarding) is untested, however. Therefore, Siberians hamsters, a natural food hoarder, were housed in a semi-natural burrow/foraging system that had (a) foraging requirement (10 revolutions/pellet), no free food (true foraging group), (b) no running wheel access, free food (general malaise control) or (c) running wheel access, free food (exercise control). We microinjected BIIE0246 (antagonist) and PYY_(3-36) (agonist) into the Arc to test the role of NPY-Y2Rs there on ingestive behaviors. Food foraging, hoarding, and intake were not affected by Arc BIIE0246 microinjection in fed hamsters 1, 2, 4, and 24 h post injection. Stimulation of NPY-Y2Rs by PYY_(3-36) inhibited food intake at 0–1 and 1–2 h and food hoarding at 1–2 h without causing general malaise or affecting foraging. Collectively, these results implicate a sufficiency, but not necessity, of the Arc NPY-Y2R in the inhibition of food intake and food hoarding by Siberian hamsters.     

Preventive effect of rikkunshito on gastric motor function inhibited by l-dopa in rats

We previously reported that ghrelin prevented l-dopa (LD)-induced inhibition of gastric emptying (GE) of a non-nutrient solution in rats. Parkinson's disease treatment involves the combined administration of l-dopa with the enzyme l-amino acid decarboxylase inhibitor, carbidopa (CD) to reduce peripheral formation of dopamine. We investigated the effect LD/CD given orogastrically (og) on GE of a non-nutrient or nutrient meal and whether og pretreatment with rikkunshito, a kampo medicine clinically used to treat gastroparesis, influenced LD/CD effect on GE and postprandial antral and duodenal motility in conscious rats. LD/CD (20/2 mg kg⁻¹) decreased significantly GE to 26.3 ± 6.0% compared to 61.2 ± 3.2% in og vehicle monitored 20-min after a non-nutrient meal and to 41.9 ± 5.8% compared to 72.9 ± 5.2% in og vehicle monitored 60 min after a nutrient meal. Rikkunshito (0.5 or 1.0 g kg⁻¹) reduced the LD/CD (20/2 mg kg⁻¹) inhibition of GE of non-nutrient meal (36.9 ± 7.4% and 46.6 ± 4.8% respectively vs. 12.1 ± 7.4% in og vehicle plus LD/CD) while having no effect alone (56.6 ± 8.5%). The ghrelin antagonist, [d-Lys³]-GHRP-6 (1 mg kg⁻¹) injected intraperitoneally partially reversed rikkunshito preventive effect on LD/CD-inhibited GE. Rikkunshito (1.0 g kg⁻¹) blocked LD/CD (20/2 mg kg⁻¹)-induced delayed GE of a nutrient meal and the reduction of postprandial antral motility. In 6-hydroxydopamine-induced Parkinson's disease rat model, rikkunshito (1.0 g kg⁻¹, og) also prevented LD/CD-inhibited gastric emptying of a nutrient meal and enhanced fasting plasma levels of acylated ghrelin. These data indicate that oral rikkunshito alleviates the delayed GE induced by LD/CD in naïve and PD rat model in part through ghrelin-related mechanisms.     

Secretory expression of functional barley limit dextrinase by Pichia pastoris using high cell-density fermentation

Heterologous production of large multidomain proteins from higher plants is often cumbersome. Barley limit dextrinase (LD), a 98 kDa multidomain starch and α-limit dextrin debranching enzyme, plays a major role in starch mobilization during seed germination and is possibly involved in starch biosynthesis by trimming of intermediate branched α-glucan structures. Highly active barley LD is obtained by secretory expression during high cell-density fermentation of Pichia pastoris. The LD encoding gene fragment without signal peptide was subcloned in-frame with the Saccharomyces cerevisiae α-factor secretion signal of the P. pastoris vector pPIC9K under control of the alcohol oxidase 1 promoter. Optimization of a fed-batch fermentation procedure enabled efficient production of LD in a 5-L bioreactor, which combined with affinity chromatography on β-cyclodextrin–Sepharose followed by Hiload Superdex 200 gel filtration yielded 34 mg homogenous LD (84% recovery). The identity of the recombinant LD was verified by N-terminal sequencing and by mass spectrometric peptide mapping. A molecular mass of 98 kDa was estimated by SDS–PAGE in excellent agreement with the theoretical value of 97419 Da. Kinetic constants of LD catalyzed pullulan hydrolysis were found to K_m,app = 0.16 ± 0.02 mg/mL and k_cat,app = 79 ± 10 s^?1 by fitting the uncompetitive substrate inhibition Michaelis–Menten equation, which reflects significant substrate inhibition and/or transglycosylation. The resulting catalytic coefficient, k_cat,app/K_m,app = 488 ± 23 mL/(mg s) is 3.5-fold higher than for barley malt LD. Surface plasmon resonance analysis showed α-, β-, and γ-cyclodextrin binding to LD with K_d of 27.2, 0.70, and 34.7 μM, respectively.     

Effect of a change in step direction from a forward to a lateral target in response to a sensory perturbation

The aim of the current study was to investigate the response of healthy older and young adults to a change in step direction from a forward to a lateral target in response to a sensory perturbation. Nine healthy older (75.1 ± 6.7 years; age range, 65–81 years) and nine young adults (27 ± 3.6 years; age range, 23–31 years) participated in the study. The sensory perturbation was a visual cue presented at random times while subjects stepped over an obstacle from quiet stance. For both young and elderly subjects there was an abrupt change in the slope of both shear ground reaction forces (GRFs) of the stance limb following the perturbation. The slope and peak of the change in GRFs was greater for the young subjects and the onset significantly earlier (205 ms compared to 271 ms). Changes in the GRFs were accompanied by an increase in bilateral gluteus medius and stance limb soleus activity. A late visual cue resulted in a delayed response for elderly subjects. These data show that a stepping response to a sensory perturbation was both delayed and of less magnitude for older adults which has implications for fall risk.     

The mechanics of activated semitendinosus are not representative of the pathological knee joint condition of children with cerebral palsy

Characteristic cerebral palsy effects in the knee include a restricted joint range of motion and forcefully kept joint in a flexed position. To show whether the mechanics of activated spastic semitendinosus muscle are contributing to these effects, we tested the hypothesis that the muscle’s joint range of force exertion is narrow and force production capacity in flexed positions is high. The isometric semitendinosus forces of children with cerebral palsy (n = 7, mean (SD) = 7 years (8 months), GMFCS levels III–IV, 12 limbs tested) were measured intra-operatively as a function of knee angle, from flexion (120°) to full extension (0°). Peak force measured in the most flexed position was considered as the benchmark. However, peak force (mean (SD) = 112.4 N (54.3 N)) was measured either at intermediate or even full knee extension (three limbs) indicating no narrow joint range of force exertion. Lack of high force production capacity in flexed knee positions (e.g., at 120° negligible or below 22% of the peak force) was shown except for one limb. Therefore, our hypothesis was rejected for a vast majority of the limbs. These findings and those reported for spastic gracilis agree, indicating that the patients’ pathological joint condition must rely on a more complex mechanism than the mechanics of individual spastic muscles.