Gonadotrophin-releasing hormone nerve terminals, tanycytes and neurohaemal junction remodelling in the adult median eminence: functional consequences for reproduction and dynamic role of vascular endothelial cells.

Although coordinated actions of several areas within the hypothalamus are involved in the secretion of gonadotrophin-releasing hormone (GnRH), the median eminence of the hypothalamus, where the nerve terminals are located, plays a particularly critical role in the release of GnRH. In adult females, prior to the preovulatory surge of GnRH, the retraction of specialised ependymoglial cells lining the floor of the third ventricle named tanycytes allows for the juxtaposition of GnRH nerve terminals with the adjacent pericapillary space of the pituitary portal vasculature, thus forming direct neurohaemal junctions. These morphological changes occur within a few hours and are reversible. Such remodelling may promote physiological conditions to enhance the central release of GnRH and potentiate oestrogen-activated GnRH release. This plasticity involves dynamic cell interactions that bring into play tanycytes, astrocytes, vascular endothelial cells and GnRH neurones themselves. The underlying signalling pathways responsible for these structural changes are comprised of highly diffusible gaseous molecules, such as endothelial nitric oxide, and paracrine communication processes involving receptors of the erbB tyrosine kinase family, transforming growth factor beta 1 and eicosanoids, such as prostaglandin E(2). Some of these molecules, as a result of their ability to diffuse within the median eminence, may also serve as synchronizing cues allowing for the occurrence of functionally meaningful episodes of GnRH secretion by coordinating GnRH release from the GnRH neuroendocrine terminals.

Birth asphyxia--analysis of 235 cases.

OBJECTIVE: To look for risk factors leading to birth asphyxia in new borns admitted in a tertiary care unit STUDY DESIGN: Retrospective analysis. SETTING: Neonatal Unit of National Institute of Child Health, Karachi from 1st January, 2001 to 31st August, 2001. PATIENTS AND METHODS: Records of 235 new borns admitted with birth asphyxia during this period were analyzed. Variables studied included antenatal care, period of gestation, place of delivery, mode of delivery, birth weight and age at arrival in the hospital. RESULTS: Majority (71%) of mothers were booked and had antenatal care, similarly most (88%) of the babies were born at term and 75.3% were delivered in maternity homes or hospitals. Caesarian sections were performed in 14% cases and rest were all vaginal deliveries. Fifty seven percent newborns weighed more than 2.5 kg and 22.5% presented to the hospital within 6 hours of birth. CONCLUSION: Birth asphyxia occurring in such a high number of booked cases delivered at term with good weight, reflects the poor perinatal services offered in those maternity homes or hospitals. It is recommended that trained personnel and neonatal resuscitation equipment should be made mandatory in all maternity homes/hospitals.

Prescribing practices of consultants at Karachi, Pakistan.

OBJECTIVE: To audit the prescribing practices of consultants. DESIGN: A randomized sample survey of 354 consultants prescription for analysis of some basic parameters as recommended by World Health Organization (WHO). MAIN OUTCOME MEASURES: To provide feedback to the prescribers for promoting rational drug therapy. RESULTS: The average number of drugs for prescription were found to be 4.51 and the average daily cost of prescribed drugs was Rs: 133.41. The antimicrobials, vitamins/minerals and injections were over-prescribed. About half of the prescribed drugs were from the National Essential Drugs List of Pakistan (NEDLP) and only a little over 12% were prescribed by their generic names. CONCLUSIONS: The prescribing practices of the consultants in Karachi are rather liberal and not rational. Overuse of antimicrobials and injections have been observed and there is general tendency of indulging in polypharmacy requiring continued medical education.

Ca2+-dependent actin remodeling in the contracting A7r5 cell.

Previous work has shown that stimulation of contraction in A7r5 smooth muscle cells with phorbol ester (PDBu) results in the disassembly and remodeling of the alpha-actin component of the cytoskeleton (Fultz et al., 2000, J Mus Res Cell Motil 21: 775-781). In the present study, we evaluated the effect of increasing intracellular calcium ion concentration [Ca2+]i by A23187 and thapsigargin on alpha- and beta-actin remodeling. The effects of A23187 and thapsigargin on cell contraction and actin remodeling were effectively identical. The two compounds caused contraction of A7r5 cells that was earlier in onset and more quickly completed than PDBu-induced contractions. Both the alpha- and beta-actin isoforms were incorporated into stress cables in the resting cell. During the interval of contraction, beta-actin cables shortened without evidence of disassembly. By comparison, the increase of [Ca2+]i resulted in partial or complete dissolution of alpha-actin cables without further remodeling. In addition, PDBu-mediated alpha-actin remodeling was blocked in the presence of A23187. Increased [Ca2+]i also caused dispersal of alpha-actinin but had no effect on the cellular distribution of talin suggesting the effect was selective for alpha-actin cytoskeletal structure. The incubation of cells in calcium-free media prevented alpha-actin dissolution by A23187/thapsigargin and also blocked PDBu-mediated remodeling. Finally, of six kinase inhibitors investigated, only ML-7 partially blocked the dissolution of alpha-actin cables by increased [Ca2+]i. The results suggest that the sustained elevation of [Ca2+]i beyond a threshold level initiates depolymerization of alpha-actin but not beta-actin. It further appears that PDBu-induced alpha-actin remodeling requires Ca2+ but increases of [Ca2+]i beyond a threshold level may inhibit this activity. The finding that ML-7 partially inhibits alpha-actin dissolution in the presence of A23187/thapsigargin may be suggesting that myosin light chain kinase (MLCK) plays a role in destabilizing alpha-actin structure in the activated cell.

Depolarized resonance light scattering by porphyrin and chlorophyll a aggregates.

A quantum mechanical model is developed for the observed resonance enhancement of light scattering by aggregates of electronically interacting chromophores. Aggregate size, monomer oscillator strength, extent of electronic coupling, and aggregate geometry are all important determinants of intensity in resonance light scattering (RLS) spectra. The theory also predicts the value of the depolarization ratio (rho(v)(90)) of RLS for a given aggregate geometry. These results are used to interpret the RLS depolarization ratios of four aggregates: tetrakis(4-sulfonatophenyl)porphine aggregated at low pH (rho(v)(90) = 0.17 at 488 nm), trans-bis(N-methylpyridinium-4-yl)-diphenylporphinato copper(II) aggregated in 0.2 M NaCl solution (rho(v)(90) = 0.13 at 450 nm) and on calf thymus DNA (rho(v)(90) = 0.20 at 454 nm), and chlorophyll a aggregates in formamide/water (rho(v)(90) = 0.23 and 0.32 at 469 and 699 nm, respectively). The analysis is consistent with a J-aggregate geometry for all four systems. Furthermore, the specific values of rho(v)(90) allow us to estimate the orientation of the monomer transition dipoles with respect to the long axis of the aggregate. We conclude that depolarized resonance light scattering spectroscopy is a powerful probe of the geometric and electronic structures of extended aggregates of strong chromophores.

pHi and serum regulate AE2-mediated Cl-/HCO3- exchange in CHOP cells of defined transient transfection status.

Anion exchanger (AE) protein-mediated anion exchange contributes to regulation of intracellular pH (pHi), Cl- concentration, and volume in vertebrate cells. We have extended the functional characterization of recombinant AE2-mediated Cl-/HCO3- exchange in single Chinese hamster ovary cells stably transfected with the polyoma large T antigen (CHOP cells) of defined transient transfection status using a novel surface marker coexpression vector. Marker expression and detection had minimal effect on the low endogenous Cl-/HCO3- exchange activity of CHOP cells, whereas coexpression of marker with AE2 elevated CHOP cell Cl-/HCO3- exchange activity 16-fold. Between pHi of 7.3 and 7.8, AE2-mediated flux of proton equivalents was activated > 11-fold by increasingly alkaline pHi without reaching saturation. This activation may be secondary to allosteric effects of pHi on AE2, in parallel with the obligatory increase in substrate intracellular HCO3- concentration. Nominal removal of CO2/HCO3- reduced AE2 activity by 90%. Addition of 10% calf serum slowly activated AE2 activity severalfold. This activation was slowly reversed after serum removal. Surface marker coexpression vectors improve both the efficiency and reliability of studies of recombinant protein function for a wide range of single cell assays in many cell types.