Met-enkephalin effects on associations formed in utero

Rat fetuses exposed to an odor stimulus and an aversive stimulus in utero showed an aversion to the odor when tested 16 days postnatally. Fetuses that also received 80 μg/kg Met-enkephalin showed a greater aversion to the odor stimulus than those subjects that did not receive the peptide. The difference between these groups was marginally significant. Control subjects did not show an aversion. But, subjects exposed to the odor and Met-enkephalin without the aversive stimulus, when tested, showed a significant preference for the odor over other control groups. These data show that associative learning in rat fetuses at 20 days of gestation may be enhanced by administration of Met-enkephalin.     

Effects of energy level and monensin on reproductive performance and lactation of beef cows

This experiment consisted of a 2 year drylot study involving 80 multiparous, suckled beef cows (40 Angus and 40 Herefords). Experimental treatments (breed, monensin and year) were arranged in 2(3) factorial to evaluate the effect of breed and monensin on reproductive performance and lactation. Cows received 85% of the NRC total digestible nutrient (TDN) requirement for the first 56 days of the 140 day trial. Cows were synchronized with Synchro-Mate B and artificially inseminated 30 days into each trial with blood samples collected for luteinizing hormone (LH) analysis from one-half of each treatment group from 24 to 36 hours after implant removal. Progesterone determinations were made on plasma samples collected at day 9 and 16 after implant removal. On day 56, milk yield estimates were obtained by the weigh-suckle-weight technique. Following these collections, energy levels were increased by allowing ad lib consumption of forage. Calves, which were removed from the cows during feeding, were given access to a 75% TDN creep ration after day 56. Milk estimates were again evaluated at 140 days. Monensin supplementation did not result in a difference in cow weight change through the restricted energy period (first 56 days) or throughout the entire 140 day period. Milk yield estimates at 56 and 140 days and calf gains throught the trial, were unaffected by monensin supplementation. Monensin resulted in no effect on conception rate or services per conception. The time of the LH peak was shifted slightly forward by monensin although not significantly. Progesterone levels were not consistently affected by monensin supplementation.     

A haemagglutination inhibition test kit for routine analysis of erythropoietin.

A commercially available haemagglutination inhibition test kit for routine analysis of erythropoietin was shown to be unable to detect elevated levels of serum Epo.     

Correlations between human portal and peripheral venous insulin and proinsulin concentrations under glucose infusion]

In 8 female patients carbohydrate tolerance was proved by means of glucose infusion test 3 days after cholecystectomy. Parameters analyzed in portal and peripheral vein blood are compared with that of 47 healthy persons. All patients demonstrate a pathological carbohydrate tolerance after cholecystectomy, further characterized by an increased lipolysis, a paradoxical rise of HGH, a diminished insulin secretion during the early and increased IRI output in the second phase. There is a significant positive correlation between portal and peripheral vein IRI concentration despite the rising portalperipheral venous IRI difference with raised portal venous IRI concentration. Corresponding differences for proinsulin concentrations can be established in the early phase only. Relations existing between blood glucose and IRI are shown by multiple regression analysis. They suggest that the altitude of IRI concentration is determined by previous blood glucose concentration.     

Results of long-term biguanide therapy as a preventive measure in protodiabetes]

55 patients with pathological glucose tolerance received a long term treatment with buformin (200 mg daily). In 43 of the protodiabetics the duration of treatment was one year, in 29 of them two years and in 11 three years. The age of the patients was 38 years and the mean relative body weight was 118 per cent. The effect of buformin on glucose tolerance and insulin secretion was tested with the glucose infusion test before and after the periods of treatment. After one year we found in 58 per cent, after two years in 69 per cent and after three years in 64 per cent of the protodiabetics an improvement of glucose tolerance. In these groups the results showed a rise of the IRI in the low responder and a decrease of the IRI in the high responder. The good effects on glucose tolerance were not demonstrable in the compared groups with long-term treatment of diet only.     

Studies of glutamate dehydrogenase. Regulation of glutamate dehydrogenase from Candida utilis by a pH and temperature-dependent conformational transition.

Glutamate dehydrogenase from Candida utilis undergoes a reversible conformational transition between an active and an inactive state at low pH AND low temperature. This conformational transition can also be followed by fluorescence measurements. The temperature-dependent equilibrium between the active and the inactive state is characterized by a transition temperature of 10.7 degrees C and a delta H value of 148 kcal/mol (620 kJ/mol). The temperature dependence of the enzymic activity above 15 degrees C yields an activation energy of 15 kcal/mol (63 kJ/mol), a larger value than that for the beef liver enzyme (9 kcal/mol; 38 kJ/mol). In contrast to the yeast enzyme the Arrhenius plot is linear and, therefore, the beef liver enzyme is not transformed into an inactive conformation at low temperatures. Sedimentation analysis shows that the inactivation of the Candida utilis enzyme is not caused by change in the quaternary structure. The pH dependence of the conformational transition at low pH measured by fluorescence change is characterized by a pK value of 7.01 for the enzyme in the absence and of 6.89 for the enzyme in the presence of 2-oxoglutarate with a Hill coefficient of 3.4 in both cases. Similar results are found when the pH dependence of the enzymic activity is analyzed. With the beef liver enzyme the same pK value is obtained but with a Hill coefficient of 1 indicating cooperativity only in the case of the Candida utilis enzyme. The best fit of the pH dependence of the rate constants of the fluorescence changes was obtained with pK values of 7.45 and 6.45 for the active and the inactive state respectively. In this model the lowest time constant which is obtained at the pH of the equilibrium was found to be 0.05 s-1. Preincubation experiments with the substrate 2-oxoglutarate but not with the coenzyme shift the equilibrium to the active conformation. The coenzyme obviously reduces the rate constant of the conformational transition. The sedimentation coefficient (SO20, w) and the molecular weight were found to be 11.0 S and 276 000, respectively. The enzyme molecule is built up by six polypeptide chains each having a molecular weight of 47 000.     

Infant neural sensitivity to dynamic eye gaze is associated with later emerging autism

Autism spectrum disorders (henceforth autism) are diagnosed in around 1% of the population [1]. Familial liability confers risk for a broad spectrum of difficulties including the broader autism phenotype (BAP) [2, 3]. There are currently no reliable predictors of autism in infancy, but characteristic behaviors emerge during the second year, enabling diagnosis after this age [4, 5]. Because indicators of brain functioning may be sensitive predictors, and atypical eye contact is characteristic of the syndrome [6-9] and the BAP [10, 11], we examined whether neural sensitivity to eye gaze during infancy is associated with later autism outcomes [12, 13]. We undertook a prospective longitudinal study of infants with and without familial risk for autism. At 6-10 months, we recorded infants' event-related potentials (ERPs) in response to viewing faces with eye gaze directed toward versus away from the infant [14]. Longitudinal analyses showed that characteristics of ERP components evoked in response to dynamic eye gaze shifts during infancy were associated with autism diagnosed at 36 months. ERP responses to eye gaze may help characterize developmental processes that lead to later emerging autism. Findings also elucidate the mechanisms driving the development of the social brain in infancy.     

Individual rotavirus-like particles containing 120 molecules of fluorescent protein are visible in living cells

Rotaviruses are large, complex icosahedral particles consisting of three concentric capsid layers. When the innermost capsid protein VP2 is expressed in the baculovirus-insect cell system it assembles as core-like particles. The amino terminus region of VP2 is dispensable for assembly of virus-like particles (VLP). Coexpression of VP2 and VP6 produces double layered VLP. We hypothesized that the amino end of VP2 could be extended without altering the auto assembly properties of VP2. Using the green fluorescent protein (GFP) or the DsRed protein as model inserts we have shown that the chimeric protein GFP (or DsRed)-VP2 auto assembles perfectly well and forms fluorescent VLP (GFP-VLP2/6 or DsRed-VLP2/6) when coexpressed with VP6. The presence of GFP inside the core does not prevent the assembly of the outer capsid layer proteins VP7 and VP4 to give VLP2/6/7/4. Cryo-electron microscopy of purified GFP-VLP2/6 showed that GFP molecules are located at the 5-fold vertices of the core. It is possible to visualize a single fluorescent VLP in living cells by confocal fluorescent microscopy. In vitro VLP2/6 did not enter into permissive cells or in dendritic cells. In contrast, fluorescent VLP2/6/7/4 entered the cells and then the fluorescence signal disappear rapidly. Presented data indicate that fluorescent VLP are interesting tools to follow in real time the entry process of rotavirus and that chimeric VLP could be envisaged as "nanoboxes" carrying macromolecules to living cells.     

Human periosteum-derived progenitor cells express distinct chemokine receptors and migrate upon stimulation with CCL2, CCL25, CXCL8, CXCL12, and CXCL13

For bone repair, transplantation of periosteal progenitor cells (PCs), which had been amplified within supportive scaffolds, is applied clinically. More innovative bone tissue engineering approaches focus on the in situ recruitment of stem and progenitor cells to defective sites and their subsequent use for guided tissue repair. Chemokines are known to induce the directed migration of bone marrow CD34(-) mesenchymal stem cells (MSCs). The aim of our study was to determine the chemokine receptor expression profile of human CD34(-) PCs and to demonstrate that these cells migrate upon stimulation with selected chemokines. PCs were isolated from periosteum of the mastoid bone and displayed a homogenous cell population presenting an MSC-related cell-surface antigen profile (ALCAM(+), SH2(+), SH3(+), CD14(-), CD34(-), CD44(+), CD45(-), CD90(+)). The expression profile of chemokine receptors was determined by real-time PCR and immunohistochemistry. Both methods consistently demonstrated that PCs express receptors of all four chemokine subfamilies CC, CXC, CX(3)C, and C. Migration of PCs and a dose-dependent migratory effect of the chemokines CCL2 (MCP1), CCL25 (TECK), CXCL8 (IL8), CXCL12 (SDF1alpha), and CXCL13 (BCA1), but not CCL22 (MDC) were demonstrated using a 96-multiwell chemotaxis assay. In conclusion, for the first time, here we report that human PCs express chemokine receptors, present their profile, and demonstrate a dose-dependent migratory effect of distinct chemokines on these cells. These results are promising towards in situ bone repair therapies based on guiding PCs to bone defects, and encourage further in vivo studies.