Route of infection that induces a high intensity of gamma interferon-secreting T cells in the genital tract produces optimal protection against Chlamydia trachomatis infection in mice

The induction of local T helper type 1 (Th1)-mediated cellular immunity is crucial for resistance of mice to genital infection by the obligate intracellular bacterium Chlamydia trachomatis. We tested the hypothesis that the route of immunization that elicits relatively high numbers of chlamydia-specific, gamma interferon (IFN-gamma)-secreting T lymphocytes (ISTLs) in the genital tract would induce optimal protective immunity against reinfection. Female BALB/c mice were infected intravaginally (i.v.), intranasally (i.n.), orally (p.o.), or subcutaneously (s.c.) with C. trachomatis. At days 7, 14, 21, and 28 postinfection, T cells isolated from the genital tract tissues were restimulated with chlamydial antigen in vitro, and the amounts of IFN-gamma induced were measured by a sandwiched enzyme-linked immunosorbent assay method. At day 7 postinfection, i.n.- and i.v.-immunized mice had high levels of chlamydia-specific ISTLs in their genital tracts (203.58 +/- 68.1 and 225.5 +/- 12.1 pg/ml, respectively). However, there were no detectable ISTLs in the genital tracts of p.o.- or s.c.-infected mice. When preinfected mice were challenged i.v. 70 days later, animals preexposed by the i.n. route were highly resistant to reinfection, with greatly reduced chlamydial burden, and suffered an attenuated infection that resolved by day 6 postchallenge. Animals preexposed by the i.v. route were modestly protected, whereas p.o. and s.c. groups were indistinguishable in this regard from control mice. The resistance of i.n.-immunized mice (and to some extent the i.v.-exposed mice) to reinfection was associated with early appearance (within 24 h) of high levels of genital ISTLs compared with mice preinfected by other routes. Furthermore, although i.n. and i.v.-immunized mice had comparable levels of chlamydia-specific immunoglobulin A (IgA) antibodies in their vaginal washes, the levels of IgG2a were four- sixfold higher in i.n.-immunized mice than in any of the other groups. The results suggested that immunization routes that foster rapid induction of vigorous genital mucosal cell-mediated immune (CMI) effectors (e.g., IFN-gamma), the CMI-associated humoral effector, IgG2a, and to some extent secretory IgA produce protective immunity against chlamydial genital infection. Therefore, i.n. immunization is a potential delivery route of choice in the development of a vaccine against Chlamydia.     

Study of the use of noninvasive ventilation of the lungs in acute respiratory insufficiency due exacerbation of chronic obstructive pulmonary disease

Noninvasive positive pressure ventilation (NPPV) is a life-saving procedure in acute respiratory failure (ARF), but its technique is not yet in routine use in many respiratory centers. We carried out a prospective randomized study comparing the combination of NPPV with conventional therapy (oxygen, bronchodilators, steroids, and theophylline) with conventional therapy alone in patients with acute respiratory failure caused by exacerbation of chronic obstructive pulmonary disease (COPD). A total of 58 patients were recruited from a large group of patients admitted to our hospital between September 1995 and March 1997. Twenty-nine patients were randomly assigned to the NPPV group and 29 to the conventional (non-NPPV) group. The patients were matched for demographic and physiological norm values (mean age 63.4 +/- 5.5 vs. 66.2 +/- 7.1 years, mean FEV1 0.68 +/- 0.15 vs. 0.74 +/- 0.16 L, PaO2 51.4 +/- 6.8 vs. 52.3 +/- 6.5 mm Hg, PaCO2 63.4 +/- 10.9 vs. 64.9 +/- 9.7 mm Hg, and pH 7.28 +/- 0.07 vs. 7.26 +/- 0.06). The outcome end points were needed for endotracheal intubation, length of hospital stay, and incidence of complications. NPPV was administered using BiPAP ventilatory device (Respironics, Inc.) by spontaneous and spontaneous/timed modes via nasal and facial masks. The mean time of NPPV was 29 +/- 25 h. Three patients refused from NPPV because of intolerance of mask or ventilation procedure. Two of them were eventually intubated and one of them died. In patients administered NPPV, we observed a significant rise of pH and fall of PaCO2 after 1 h of ventilation, in contrast to the non-NPPV group (7.34 +/ 0.09 vs. 7.21 +/- 0.08, p < 0.05; 53.2 +/- 10.7 vs. 71.4 +/- 10.2 mm Hg, p < 0.01, respectively). The need in intubation was lower in the NPPV group as compared to the reference group (12 vs. 28%, p = 0.18), mortality rate was higher in the non-NPPV group (31 vs. 8%, p = 0.03), and hospital stay was shorter in NPPV patients (26 +/- 7 vs. 34 +/- 10 days). The incidence of complications was lower in the NPPV group, they were less significant, and did not involve discontinuation of ventilation. Hence, NPPV is a first-line therapy in patients with ARF caused by COPD exacerbation, due to obvious advantages over conventional methods of treatment.     

Can we use erythrocytes for the study of the activity of the ubiquitous Na+/H+ exchanger (NHE-1) in essential hypertension?

Both Na+/Li+ countertransport and electrochemical proton gradient (delta mu(H+))-induced Na+ and H+ fluxes are increased in erythrocytes from patients with essential hypertension. It was assumed that these abnormalities are related to ubiquitous (housekeeping) forms of the Na+/H+ exchanger (NHE-1). To examine this hypothesis, we compared kinetic and regulatory properties of erythrocyte Na+/Li+ countertransport and delta mu(H+)-induced Na+ and H+ fluxes with data obtained for cloned isoforms of the Na+/H+ exchanger. In human erythrocytes, Na+/Li+ countertransport exhibited a hyperbolic dependence on [Na+]0 with a K0.5 of approximately 30 to 40 mmol/L. The activity of this carrier was increased by two-fold in the fraction of erythrocytes enriched with the old cells, was inhibited by 0.1 mmol/L phloretin, and was insensitive to both 1 mmol/L amiloride and ATP depletion. In contrast, delta mu(H+)-induced 22Na influx was exponentially increased at [Na+]0 > 60 mmol/L, was insensitive to phloretin, was partly decreased by both 1 mmol/L amiloride and ATP depletion, and was the same in total erythrocytes and in the old cells. The values of Na+/Li+ countertransport and delta mu(H+)-induced Na+ influx in erythrocytes from different species were not correlating and their ratio in human, rat, and rabbit erythrocytes was 10:1:170 and 1:5:1 for Na+/ Li+ countertransport and delta mu(H+)-induced Na+ influx, respectively. In contrast to the majority of nonepithelial cells and cells transfected with an ubiquitous isoform of Na+/H+ exchanger, both delta mu(H+)-induced Na+ influx and Na+/Li+ countertransport in human erythrocytes were completely insensitive to ethylisopropyl amiloride (20 micromol/L) and cell shrinkage. Thus, our data strongly suggest that human erythrocyte Na+/Li+ countertransport and delta mu(H+)-induced Na+/H+ exchange are mediated by the distinct transporters. Moreover, because the properties of these erythrocyte transporters and NHE-1 are different, it complicates the use of erythrocytes for the identification of the mechanism for activating the ubiquitous form of Na+/H+ exchanger in primary hypertension.     

Epidemiologic data of stroke. Data of the WHO-MONICA Project in Germany

Analyses of stroke morbidity or mortality are usually based on official statistics. A reduction in stroke mortality rates has been shown for many countries. It is not clear, however, whether this is due to declining morbidity or case fatality (or both). For this purposes population-based register data are required. Using the standardized methodology of the WHO-MONICA Project, stroke cases were also registered in Germany from 1984 to 1993 (7,435 first-ever and recurrent stroke cases). The data collection was almost restricted to East Germany. The age-specific stroke rates in males/females showed an increase from 9/11 per 100,000 population in the youngest age group (25-34) to 1,005/779 cases per year in the oldest group under study (65-74). If one tries to classify stroke types, which is not always possible in a population-based register, the best estimate for men (women) would be: 63(62%) thromboembolic stroke, 25(22)% intracerebral hemorrhage, and 12(17)% subarachnoid hemorrhage. The 28-day case fatality of the 25-74 year old stroke patients was found to be about 40%. Neither for stroke attacks nor for case fatality was a convincing time trend over the 10-year period found. The very small changes observed over 10 years time should lead to increased attention to strokes, particularly primary and secondary prevention, and this not only in East Germany. This applies also for treatment in the acute phase, because the case fatality before admission in the hospital and during the first few days is still very high. Population-based studies of the long-term prognosis of stroke patients in Germany are also missing, i.e., including the effectiveness of various forms of treatment and rehabilitation. Systematic monitoring of the development in this field is an important part of the assessment of the quality and effectiveness of the health care service.     

Endosulfan poisoning in Northern India: a report of 18 cases

Eighteen cases of endosulfan poisoning by accidental overexposure during spray, admitted between October 1995 to September 1997, were observed and analyzed. These accounted for approximately one third of the total number of poisoning cases admitted in our unit during this period. Nausea, vomiting abdominal discomfort, tonic and clonic convulsions, confusion, disorientation, and muscular twitchings were cardinal manifestations. None of the patients succumbed to their illness. Analysis of various incriminating factors revealed that accidental overexposure was due to failure to adhere to the instructions for spray either due to ignorance or due to illiteracy. All the patients avoided preventive measures and developed toxicity both due to inhalation and absorption through skin. Endosulfan (a chlordiene derivative) poisoning is gaining up momentum in this part of world and has become an important matter for public health in India.     

Glyceryl nonivamide: a capsaicin derivative with cardiac calcitonin gene-related peptide releasing, K+ channel opening and vasorelaxant properties

In this study, the aorta vasorelaxant, coronary calcitonin gene-related peptide (CGRP) releasing, and atrial contractility effects of glyceryl nonivamide (GLNVA) were investigated in guinea pigs. In the isolated thoracic aorta, although GLNVA (0.01-50 microM) concentration dependently induced endothelium-independent relaxations and relaxed phenylephrine-(1.0 microM) induced contractions, it failed to relax 80 mM KCI-induced contractions. The GLNVA (1.0 microM) relaxation response in the aorta was significantly inhibited by tetraethylammonium (2.5-10 mM) or ouabain (5.0 microM) and was attenuated by increased extracellular potassium gradient (10-30 mM). Glibenclamide (0.01-10 microM) dose dependently antagonized the GLNVA relaxant effect. In the isolated perfused guinea pig heart, GLNVA (0.1-10 microM) increased CGRP-like immunoreactivity outflow from coronary circulation in a concentration-dependent manner. In the isolated right and left guinea pig atria, GLNVA (0.01-10 microM) produced concentration-dependent positive inotropic and chronotropic effects, but these effects were inhibited by pretreatments with ruthenium red (1.0 microM), capsazepine (10 microM), human calcitonin-gene-related peptide (CGRP(8-37)) (1.0 microM) and sensory neuron denervation, respectively. Based on these findings, we suggest that CGRP may be released by GLNVA from cardiovascular sensory neuron, and it then activates CGRP receptors on the coronary artery and atrium. The GLNVA-induced vasorelaxant effect in the vascular smooth muscle of the aorta is due to CGRP release associated K+ channel opening, and this effect eliminates capsaicin-derived excitability-associated K+ channel blocking activities.     

The crystal and molecular structure of a valinomycin analogue cyclo[(D-Val-L-Lac-L-Ala-D-Hyi)2(D-Val-L-Lac-L-Val-D-Hyi)]. H2O(C50H82N6 O18.H2O)

The crystal and molecular structure of the valinomycin analogue, cyclo[(D-Val-L-Lac-L-Ala-D-Hyi)2(D-Val-L-Lac-L-Val-D-Hyi)] has been solved by x-ray direct methods using the "Shake and Bake" procedure. The crystals, grown from a mixture of octane/CH2Cl2, belong to space group P2(1) (Z = 4) with cell parameters a = 10.29, b = 32.08, c = 18.73 A, beta = 97.05 degrees, and contain two molecules per asymmetric unit. After anisotropic refinement the standard reliability factor was Rl = 0.058. The conformations of both independent molecules is similar to that observed for isoleucinomycin, cyclo[-(D-Ile-L-Lac-L-Ile-D-Hyi)3] [V. Z. Pletnev et al. (1980) Biopolymers, Vol. 19, pp. 1517-1534]. The structure has an asymmetric conformation stabilized by six intramolecular H bonds, five bonds being of the 4-->1 type and one bond being of the 5-->1 type. One water molecule is caged in the internal cavity of each cyclodepsipeptide. This conformation could represent an intermediate state between free and complexed forms of valinomycin.