Searches among mycobacterial cultures for antileprosy vaccines.

All mycobacteria species share some antigens, so there may be cultivable mycobacterial cultures that can provide vaccine protection against leprosy. Vaccine protection against Mycobacterium leprae infections in mice has been demonstrated for M. leprae itself, as living or heat-killed suspensions, and for Mycobacterium bovis (BCG), as living suspensions. Results are reported here with 17 other cultures. The mycobacterial suspensions were injected intradermally, and the mice were challenged in the footpad with infectious suspensions of M. leprae. In two experiments the mice were also challenged by footpad injections of 10(7) heat-killed M. leprae so the footpad enlargment could be measured. That some mycobacterial suspensions were immunogenic for some of their own antigens was suggested by reactions at the vaccine site and enlargement of the regional lymph nodes. Some mycobacterial suspensions also stimulated footpad enlargement on challenge by homologous suspensions or by challenge with M. leprae suspensions. Consistent protection against infectious challenge with M. leprae was observed only with BCG and M. leprae, however.     

Hereditary Characteristic that Varies Among Isolates of Mycobacterium leprae

Isolates of Mycobacterium leprae in mouse foot pads were found to differ in two related properties, the average rate of growth between inoculation and harvest (G) and the number of bacilli in the harvest (H). For “fast” strains the median values for G were less than 25 days per generation, and the median values for H were above 10^6.1. For “slow” strains the median values for G were above 30, and the median values for H were below 10^5.6. The G and H values for the 59 isolates for which data were available formed a continuous spectrum between the two extremes; there was no correlation with dapsone resistance. The fastness characteristic was stable; it did not change on passage in mice and was in agreement when more than one isolate had been made from the same patient. No important differences were apparent according to geographic origin of the infection of the patient. Histological studies showed that fast strains grew to a higher level without inducing the infiltrate of lymphocytes and macrophages that appears at the end of the logarithmic phase of growth in mouse foot pads. Although fast strains often had higher ratios of solidly staining (and presumably viable) bacilli in the inoculum, the fast-slow difference was not accounted for by the solid ratio. Slow strains differed from fast by having longer times until harvest and by having fewer generations of growth, even when their frequently lower solid ratios were taken into account.     

Non-secretion of mutant proteins of the glaucoma gene myocilin in cultured trabecular meshwork cells and in aqueous humor

Until recently, very little was known about the molecular mechanisms responsible for the development of glaucoma, a leading cause of blindness worldwide. Mutations in the glaucoma gene myocilin (MYOC, GLC1A) are associated with elevated intraocular pressure and the development of autosomal dominant juvenile glaucoma and a subset of adult-onset glaucoma. MYOC is expressed in the trabecular meshwork (TM), a tissue responsible for drainage of aqueous humor from the eye, and the tissue involved in elevated intraocular pressure associated with glaucoma. To better understand the role of MYOC in glaucoma pathogenesis, we examined the expression of normal and mutant myocilin in cultured ocular (TM) and non-ocular cells as well as in the aqueous humor of patients with and without MYOC glaucoma. Normal myocilin was secreted from cultured cells, but very little to no myocilin was secreted from cells expressing five different mutant forms of MYOC. In addition, no mutant myocilin was detected in the aqueous humor of patients harboring a nonsense MYOC mutation (Q368X). Co-transfection of cultured cells with normal and mutant myocilin led to suppression of normal myocilin secretion. These studies suggest that MYOC glaucoma is due either to insufficient levels of secreted myocilin or to compromised TM cell function caused by congestion of the TM secretory pathway.     

Role of nitric oxide in the biology, physiology and pathophysiology of reproduction

Following its benchmark discovery, nitric oxide (NO) is nowknown to play important functional roles in a variety of physiologicalsystems. Within the vasculature, NO induces vasodilation, inhibitsplatelet aggregation, prevents neutrophil/platelet adhesionto endothelial cells, inhibits smooth muscle cell proliferationand migration, regulates programmed cell death (apoptosis) andmaintains endothelial cell barrier function. NO generated byneurons acts as a neurotransmitter, whereas NO generated bymacrophages in response to invading microbes acts as an antimicrobialagent. Because neurons, blood vessels and cells of the immunesystem are integral parts of the reproductive organs, and inview of the important functional role that NO plays in thosesystems, it is likely that NO is an important regulator of thebiology and physiology of the reproductive system. Indeed, inthe past 10 years, NO has established itself as a polyvalentmolecule which plays a decisive role in regulating multiplefunctions within the female as well as the male reproductivesystem. This review provides an overview of the role of NO invarious reproductive organs under physiological and pathologicalconditions.     

Comparative analysis of the polycystic kidney disease 1 (PKD1) gene reveals an integral membrane glycoprotein with multiple evolutionary conserved domains

PKD1 is the major locus of the common genetic disorder autosomal dominant polycystic kidney disease (ADPKD). Analysis of the predicted protein sequence of the human PKD1 gene, polycystin, shows a large molecule with a unique arrangement of extracellular domains and multiple putative transmembrane regions. The precise function of polycystin remains unclear with a paucity of mutations to define key structural and functional domains. To refine the structure of this protein we have cloned the genomic region encoding the Fugu PKD1 gene. Fugu PKD1 spans 36 kb of genomic DNA and has greater complexity with 54 exons compared with 46 in man. Comparative analysis of the predicted protein sequences shows a lower level of homology than in similar studies with identity of 40 and 59% similarity. However key structural motifs including leucine rich repeats (LRR), a C-type lectin and LDL-A like domains and 16 PKD repeats are maintained. A region of homology with the sea urchin REJ protein was also confirmed in Fugu but found to extend over 1000 amino acids. Several highly conserved intra- and extra- cellular regions, with no known sequence homologies, that are likely to be of functional importance were detected. The likely structure of the membrane associated region has been refined with similarity to the PKD2 protein and voltage gated Ca2+ and Na+ channels highlighted over part of this area. The overall protein structure has therefore been clarified and this comparative analysis derived structure will form the basis for the functional study of polycystin and its individual domains.      

Blockade of SK-type Ca2+-activated K+ channels uncovers a Ca2+-dependent slow afterdepolarization in nigral dopamine neurons

Sharp electrode current-clamp recording techniques were used to characterize the response of nigral dopamine (DA)-containing neurons in rat brain slices to injected current pulses applied in the presence of TTX (2 microM) and under conditions in which apamin-sensitive Ca2+-activated K+ channels were blocked. Addition of apamin (100-300 nM) to perfusion solutions containing TTX blocked the pacemaker oscillation in membrane voltage evoked by depolarizing current pulses and revealed an afterdepolarization (ADP) that appeared as a shoulder on the falling phase of the voltage response. ADP were preceded by a ramp-shaped slow depolarization and followed by an apamin-insensitive hyperpolarizing afterpotential (HAP). Although ADPs were observed in all apamin-treated cells, the duration of the response varied considerably between individual neurons and was strongly potentiated by the addition of TEA (2-3 mM). In the presence of TTX, TEA, and apamin, optimal stimulus parameters (0.1 nA, 200-ms duration at -55 to -68 mV) evoked ADP ranging from 80 to 1,020 ms in duration (355.3 +/- 56.5 ms, n = 16). Both the ramp-shaped slow depolarization and the ensuing ADP were markedly voltage dependent but appeared to be mediated by separate conductance mechanisms. Thus, although bath application of nifedipine (10-30 microM) or low Ca2+, high Mg2+ Ringer blocked the ADP without affecting the ramp potential, equimolar substitution of Co2+ for Ca2+ blocked both components of the voltage response. Nominal Ca2+ Ringer containing Co2+ also blocked the HAP evoked between -55 and -68 mV. We conclude that the ADP elicited in DA neurons after blockade of apamin-sensitive Ca2+-activated K+ channels is mediated by a voltage-dependent, L-type Ca2+ channel and represents a transient form of the regenerative plateau oscillation in membrane potential previously shown to underlie apamin-induced bursting activity. These data provide further support for the notion that modulation of apamin-sensitive Ca2+-activated K+ channels in DA neurons exerts a permissive effect on the conductances that are involved in the expression of phasic activity.     

Immunochemical cross-reactivity between the dissociated capsid proteins of PVY group plant viruses

Primary (4–5-week) and long-term (12-week) antisera were elicited against the pyrrolidine-dissociated capsid proteins (D-proteins) of tobacco etch virus (TEV) and potato virus Y (PVY), and subsequently used to ascertain the degree of immunochemical cross-reactivity between several PVY group viral D-proteins. Primary antisera were highly specific for homologous antigen. However, immunodiffusion and cross-absorption experiments with long-term anti-TEV D-protein and anti-PVY D-protein sera revealed pronounced cross-reactivity among the D-proteins of 14 different PVY group viruses. Furthermore, all cross-reactions involved the same antibody population except in the case of celery mosaic virus (CeMV) D-protein where but a portion of the antibody population was reactive. Spur formation in immuno-diffusion experiments was only observed among heterologous D-proteins when they were either adjacent to homologous D-protein antigen or (in an opposite direction) when compared with CeMV D-protein. Wheat streak mosaic virus dissociated protein, in contrast, was not found to be cross-reactive with PVY group D-proteins in reciprocal tests, and no cross-reactions were observed with the dissociated proteins of 6 additional viruses outside the PVY group.     

Recommendations on performance characteristics of diagnostic exposure meters: report of AAPM Diagnostic X-Ray Imaging Task Group No. 6.

Task Group 6 of the Diagnostic X-Ray Imaging Committee of the American Association of Physicists in Medicine (AAPM) was appointed to develop performance standards for diagnostic x-ray exposure meters. The recommendations as approved by the Diagnostic X-Ray Imaging Committee and the Science Council of the AAPM are delineated in this report and provide specifications on meter precision, calibration accuracy, calibration reference points, linearity, energy dependence, exposure rate dependence, leakage, amplification gain settings, directional dependence, the stem effect, constancy checks, and calibration intervals. The report summarizes recommendations for meters used in mammography, general purpose radiography including special procedures, computed tomography, and radiation safety surveys for x-ray radiography.     

Attenuation of ethanol tolerance by a novel stimulus

A Pavlovian conditioning model of tolerance emphasizes that an association between predrug cues and the systemic effects of the drug contributes to tolerance. On the basis of this model, established tolerance should be attenuated by external inhibition, i.e., by presentation of a novel, extraneous stimulus. This prediction was evaluated in the present experiment. Rats that were so tolerant to the hypothermic effect of ethanol that they evidenced no drug-induced decrease in temperature were presented with a bright strobe light following ethanol administration. The light precipitated a large decrease in temperature in these rats. These results provide further evidence that tolerance to the hypothermic effect of ethanol is, in part, mediated by learning.     

RENAL TRANSPLANTATION – AN EARLY EXPERIENCE

Renal transplant (RT) is now a therapy of choice for end stage renal disease (ESRD). The Nephrology Unit, Asvini started functioning in Dec 90 and to date 1298 sittings of hemodialysis have been given to 45 patients. Of these, 35 were in ESRD and 11 patients underwent renal transplantation at this hospital during the period Jan 91 – Dec 93. One patient expired after 18 months of transplantation due to infection. Early experience in screening patients for RT, use of immunosuppression, management of rejection episodes and protocol are presented with special emphasis on its relevance to the Armed Forces.KEY WORDS: Transplantation, Renal Failure, Immunosuppression, Rejection