Calcium-evoked dendritic exocytosis in cultured hippocampal neurons. Part II: mediation by calcium/calmodulin-dependent protein kinase II.

Calcium-evoked dendritic exocytosis (CEDE), demonstrated in cultured hippocampal neurons, is a novel mechanism that could play a role in synaptic plasticity. A number of forms of neuronal plasticity are thought to be mediated by calcium/calmodulin-dependent protein kinase II (CaMKII). Here, we investigate the role of CaMKII in CEDE. We find that the developmental time course of CEDE parallels the expression of alphaCaMKII, a dominant subunit of CaMKII. An inhibitor of this enzyme, KN-62, blocks CEDE. Furthermore, 7 d in vitro neurons (which normally do not express alphaCaMKII nor show CEDE) can undergo CEDE when infected with a recombinant virus producing alphaCaMKII. Expression of a constitutively active CaMKII produces dendritic exocytosis in the absence of calcium stimulus, and this exocytosis is blocked by nocodazole, an inhibitor of microtubule polymerization that also blocks CEDE. These results indicate that CEDE is mediated by the activation of CaMKII, consistent with the view that CEDE plays a role in synaptic plasticity

A study of feto-maternal outcome in cases of pre-labour rupture of membranes

Background: Prelabour rupture of membranes is a common obstetric problem and the assessment of women with possible membrane rupture is a management issue faced in every day practice. The diagnosis and treatment of Prelabour rupture of membranes depends upon the facilities available. Unrecognized and inadequately treated conditions can lead to various maternal and neonatal complications.Methods: The present prospective study was conducted on 200 pregnant women satisfying the inclusion criteria after getting consent for participation. Pregnant women between the gestational age of 28 – 40 weeks who presented with Prelabour rupture of membranes to casualty were enrolled into the study.Results: 76.5% of the study participants were between 37-40 weeks of gestation and the remaining 23.5% belonged to the preterm population. Maternal morbidity was found to be 68% among the study participants with latency period more than 24 hrs. 37.5% of the neonates developed morbidity and the neonatal mortality was 2%.Conclusions: Prelabour rupture of membranes contributes significantly to maternal morbidity, neonatal morbidity and perinatal mortality

Relevance of immuno-contraceptive vaccines for population control. I. Hormonal immunocontraception

Human chorionic gonadotropin--subunit β (β-hCG) has been so far most extensively studied antigen for immuno-contraceptive properties. Studies of vaccination of non-human primates has been so far controversially reported since some antisera obtained from baboons immunized with β-hCG had shown cross-reactivity to other tissues. Despite these problems and concerns about the complete safety of this first generation vaccine, a decision has been made to proceed with a limited clinical trials with this vaccine. Results from these trials are encouraging and indicate that contraceptive antibody titers can be achieved with permissible adjuvants. Neutralization of LH and/or FSH by circulating antibodies may impede their action and interfere with the maturation of gametes. Gonadotropin releasing hormone can be also a suitable target for immunological attack. The human reproductive process entails numerous possible sites for immunological intervention aimed at controlling fertility. Currently under investigation are a vaccine based on gonadotropin releasing hormone (GnRH) interception, regulation of male fertility by immunointerception of follicle-stimulating hormone (FSH), vaccines based on the neutralization of human chorionic gonadotropin (hCG), and anti-hCG immunization. In Phase I clinical trials, passive transfer of anti-GnRH antibody has produced short-term infertility; long-term infertility has been achieved through active immunization against the decapeptide. In terms of FSH, the research has failed to demonstrate how long-term neutralization brings about male infertility. Before immunization against FSh is considered further as a means of immunocontraception, further studies are needed on the required levels of physiological FSH and testosterone for normal testicular functions to be maintained. 4 other Phase I clinical trials in this area involve vaccines based on the β subunit of hCG or its fragment. Preliminary results from these trials suggest that contraceptive antibody titers can be obtained with permissible adjuvants, especially tetanus toxoid

Infection of frog neurons with vaccinia virus permits in vivo expression of foreign proteins

Vaccinia virus can be used to infect cells in the CNS of frogs, Xenopus laevis, and Rana pipiens, both in vivo and in vitro. In vivo infections were accomplished by injection of viral solution into the tectal ventricle of stage 40-48 tadpoles or by local injections into distinct neural regions. Infections with high titer of virus injected into the ventricle resulted in the majority of cells in the brain expressing foreign protein, while cells in the retina and optic nerve showed no expression. Infection with lower viral titers resulted in fewer infected cells that were distributed throughout the otherwise normal tissue. Intense expression of foreign protein in the brain was observed 36 hr after injection and remained high for at least 4 days. Infected animals developed normally and had the same number of cells in the optic tectum as control animals. Infection with a recombinant virus carrying the gene for Green Fluorescent Protein labels neurons, so that infected cells can be observed in vivo. Vaccinia virus provides a versatile means to alter proteins in distinct populations of neurons in amphibia

Vaccinia virus transfection of hippocampal slice neurons

Here we describe a technique that uses a recombinant vaccinia virus to transfect neurons in rat hippocampal slices. This technique allows the use of molecular biological manipulations on neuronal tissue while maintaining intact synaptic function. This method should be useful in testing specific hypotheses regarding the role of synaptic proteins

Characteristics of monoclonal antibodies against porcine zona pellucida-3 and their functional relevance

Seven monoclonal antibodies (MAs) against 55 kDa glycoprotein family of porcine zona pellucida (ZP3) reacting with either ZP3 α (MA-7, MA-27, MA-28) or ZP3 β (MA-1, MA-2, MA-11, MA-30) have been described. MA-1, -2, -27, -28 and -30 do not recognize carbohydrate determinants as shown by their reactivity to the deglycosylated (DG) ZP3 alpha and ZP3 beta. Indirect immunoperoxidase studies showed that all MAs reacted with zona pellucida from porcine and monkey ovaries. Only MA-1 and -27 reacted with ZP from rabbit ovary as well, while none of the MAs recognised mouse ZP, MA-7, -11, -27, -28 and -30 inhibited in vitro, the zona lysis by trypsin as well as the binding of ZP3 to sperm membrane vesicle as investigated by ELISA