Functional properties of recombinant GABAA receptors composed of single or multiple β subunit subtypes

GABA_A receptor (GABAR) isoforms in the central nervous system are composed of combinations of α(1–6), β(1–4), γ(1–4), δ(1) and (1) subunit subtypes arranged in a pentamer. Many regions of the brain express high levels of mRNA encoding several different subunits and even multiple subunit subtypes. The stoichiometry of GABAR isoforms is unclear, and the number and identity of individual subunit subtypes that are coassembled remain uncertain. To examine the role of β subunit subtypes in the functional properties of GABARS and to determine whether multiple β subtypes can be coassembled in functional GABARs, plasmids containing cDNAs encoding rat β1 and/or β3, α5 and γ2L subtypes were cotransfected into L929 fibroblasts. The properties of the expressed receptor populations were determined using whole-cell and single-channel recording techniques. The α5β1γ2L isoform was less sensitive to GABA than the α5β3γ2L isoform. α5β1γ2L isoform currents were also insensitive to the allosteric modulator loreclezole, while α5β3γ2L isoform currents were strongly potentiated by loreclezole. Fibroblasts transfected with plasmids containing cDNAs for both β1 and β3 subtypes along with α5 and γ2L subtypes produced a receptor population with an intermediate sensitivity to GABA which was insensitive to loreclezole. These results suggest that functional GABARs can be formed that contain two different β subunit subtypes with properties different from receptors that contain only a single β subtype and that the β subunit subtypes influence the response of GABARs to GABA and to the allosteric modulator loreclezole.     

Low-Calcium Diets Enhance Phytate-Phosphorus Availability

This study showed that, in chickens, the negative effects of phytate-phosphorus in a low inorganic phosphorus diet could be completely reversed through the additive effects of reduced dietary calcium and increased cholecalciferol. In the future, perhaps greater reliance on more readily available plant phosphorus sources may be instituted in developing countries, where cheap sources of inorganic phosphorus are difficult to obtain.     

Lack of involvement of reactive oxygen in the cytotoxicity of mitoxantrone,CI941 and ametantrone in MCF-7 cells: comparison with doxorubicin

Summary The MCF-7 cell S9 fraction and whole MCF-7 cells can mediate one-electron-redox cycling of doxorubicin, giving rise to concomitant oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH), formation of a drug semiquinone free radical, consumption of molecular oxygen and formation of superoxide anions and hydroxyl radicals. Doxorubicin redox cycling was consistent with DNA strand breakage and cell kill in MCF-7 cells. In contrast, no evidence for redox cycling was found for mitoxantrone (MIT), CI941 or ametantrone (AMET) in MCF-7 cells. Despite the absence of redox cycling, the CI941, MIT, and AMET concentrations resulting in 50% mortality (LC₅₀; 1.5×10^–10, 5.2×10^–9 and 1.2×10^–6 m, respectively) of MCF-7 cells were lower than that of DOX (3.0×10^–6 m). Furthermore, the higher cytotoxicity of MIT and CI941 as compared with AMET or DOX was associated with greater efficiency in inducing DNA strand breakage in MCF-7 cells as determined by alkaline elution. Sine MIT and CI941 proved to be the most potent DNA-damaging and cytotoxic agents in this study, the ability of DOX to undergo redox cycling does not appear to confer increased cytotoxic potential on this agent. The present study revealed several important aspects with regards to the structural modification of anthraquinone antitumour agents. Firstly, the C1 and C4 postitioning of the hydroxyethylamino side chains on MIT, CI941 and AMET is associated with a lack of flavin reductase-mediated activation of these agents. Secondly, the possession of a C5 or C8 aromatic hydroxyl group appears to be intimately involved in the enhanced DNA strand breakage and cytotoxic potency of MIT and CI941, since AMET does not possess these groups. These findings indicate that future development of quinone antitumour agents should concentrate on compounds that do not undergo redox cycling but do possess aromatic hydroxyl groups, since the latter appear to be responsible for the enhanced cytotoxicity of MIT and CI941.Abbreviations DOX doxorubicin - MIT mitoxantrone - AMET ametantrone - SOD superoxide dismutase - DMPO 5,5-dimethyl-1-pyrroline-N-oxide - ESR electron spin resonance - SSF strand scission factor This work was funded by the Cancer Research Campaign (UK)     

A comparison of preoperative and intraoperative evaluation of patients undergoing pelvic reconstructive surgery for pelvic organ prolapse using the pelvic organ prolapse quantification system

OBJECTIVE: To compare the pre- and intraoperative situation using the POP-Q system during optimally standardized conditions of both examinations. STUDY DESIGN: In a prospective observational study, 108 women were compared. The POP-Q in the outpatient department (preoperative) was compared with the situation just prior to surgery after full anesthesia was reached (intraoperative). During the intraoperative measurement, traction with 0.5 kg force was applied on all relevant places. RESULTS: The pre- and intraoperative measurements were all significant correlated with the R-values between 0.43 and 0.85. All six points, which are measured during the POP-Q, were more prolapsed in the intra- as compared with the preoperative situation. The points Bp, C, and D were significantly more prolapsed, but for the points Aa, Ba, and Ap this was not significant. Fifteen patients were upstaged by the intraoperative measurements and five patients were downstaged in the overall POP-Q grading system. CONCLUSIONS: Intraoperative evaluation of the prolapse can reveal significant changes as compared with the preoperative situation. In general, the prolapse is more pronounced especially in the middle and posterior compartment.     

Vein of Galen aneurysm presenting with recurrent aseptic meningitis and subsequent spontaneous thrombosis

Clinical and laboratory findings are described in the case of a patient with a vein of Galen aneurysm who presented with recurrent aseptic meningitis for which no etiology could be identified. The patient subsequently developed thalamic and intraventricular hemorrhage associated with partial thrombosis of the dilated vein of Galen. Review of the literature revealed no previously reported association of these conditions. Recurrent cerebral venous thrombosis involving the fistula is hypothesized as the cause of repeated inflammatory reactions near the subarachnoid space. More extensive thrombosis may then have precipitated the hemorrhage.     

Emergence of occult minority genotype 2b hepatitis C infection in an HIV-1-co-infected patient treated for genotype 5a HCV infection with 48 weeks of pegylated-interferon-alpha 2b and ribavirin.

An HIV-1/hepatitis C virus (HCV) co-infected patient with haemophilia received a 48-week course of pegylated interferon-alpha-2b and ribavirin therapy for genotype 5a HCV infection. Virological response was achieved at week 24. At the end of treatment, HCV RNA in serum was detected and identified to belong to genotype 2b, rather than genotype 5a. A sensitive method for identifying minority HCV genotypes in pre-treatment serum showed genotype 2b HCV carriage prior to treatment. Sequencing the interferon sensitivity-determining region of the HCV NS5A gene obtained from pre-, intra- and post-treatment sera revealed emergence of quasispecies bearing R-->K and M-->A/T mutations at codons 2222 and 2223, respectively. Occult presence of minority HCV subpopulations and their acquisition of mutations following therapy can result in poor treatment outcome.     

The use and cost of HIV service provision in England in 1996. National Prospective Monitoring System (NPMS) Steering Group and NPMS Working Party on Costs.

OBJECTIVE: The aim of the study was to measure the use and estimate the cost of HIV service provision in England. DESIGN AND SETTING: Standardised activity and case-severity data were collected prospectively in 10 English HIV clinics (5 London and 5 non-London sites) for the periods 1 January 1996 to 30 June 1996 and 1 July 1996 to 31 December 1996 and linked to unit cost data. In total, 5440 patients with HIV infection attended during the first 6 months and 5708 during the second 6 months in 1996. MAIN OUTCOME MEASURES AND RESULTS: The mean number of inpatient days per patient-year for patients with AIDS was 19.7 [95% confidence interval (CI): 13.7 to 25.7] for January to June and 20.8 (95% CI: 15.3 to 26.4) for July to December 1996. The mean number of outpatient visits for asymptomatic patients with HIV infection was 14.8 (95% CI: 11.9 to 17.6) and 13.3 (95% CI: 10.8 to 15.7) for the respective periods and 16.1 (95% CI: 13.21 to 18.97) and 15.7 (95% CI: 11.2 to 20.2), respectively, for patients with symptomatic non-AIDS (i.e. symptomatic patients with HIV infection but without AIDS-defining conditions). Substantial centre-to-centre variation was observed, suggesting that many clinics can continue the shift from an inpatient- to an outpatient-based service. Cost estimates per patient-year for HIV service provision for 1996 varied from 4695 Pounds (95% CI: 3769 Pounds to 5648 Pounds) for asymptomatic patients, to 7605 Pounds (95% CI: 6273 Pounds to 8909 Pounds) for symptomatic non-AIDS patients to 20,358 Pounds (95% CI: 17,681 Pounds to 23,206 Pounds) for patients with AIDS. CONCLUSIONS: Different combinations of antiretroviral therapy affect the cost estimates of HIV service provision differently. Anticipated reduction in inpatient-related activity through the increased use of combination antiretroviral therapy will further shift service provision from an inpatient- to outpatient-based service and reduce costs per patient-year. The extent and duration of such effects are currently unknown. The long term effects of combination treatment on the morbidity and mortality patterns of individuals infected with HIV are also currently unknown, as are their implications on the use and cost of HIV service provision. Multicentre databases like the National Prospective Monitoring System (NPMS) will provide healthcare professionals with information to improve existing services and anticipate the impact of new developments.     

Photons, physiatrics, and physicians: a practical guide to understanding laser light interaction with living tissue, part I

In the past 32 years, lasers have progressed from interesting curiosities to important instruments in medicine and surgery, with a wide variety of wavelengths and medical-surgical applications. Today's physician is faced with a bewildering array of laser types, each touted by its manufacturer as the ultimate surgical tool. This article sets forth in simple, understandable prose, the basic principles of the interaction of laser light with living tissue, so that the physician can decide which type of laser is best suited to a given application, without relying on the manufacturer's sales literature. The topics discussed are the nature of light; reflection, absorption, scattering, and attenuation in living tissue; physical processes by which laser light destroys tissue; relative importance of the three unique properties of laser light in surgery and therapy; temporal modes of lasers; means of delivering laser beams to their targets; and considerations in the selection of laser systems.