Experimental cannabinoid 2 receptor-mediated immune modulation in sepsis.

Sepsis is a complex condition that results from a dysregulated immune system in response to a systemic infection. Current treatments lack effectiveness in reducing the incidence and mortality associated with this disease. The endocannabinoid system offers great promise in managing sepsis pathogenesis due to its unique characteristics. The present study explored the effect of modulating the CB2 receptor pathway in an acute sepsis mouse model. Endotoxemia was induced by intravenous injection of lipopolysaccharide (LPS) in mice and intestinal microcirculation was assessed through intravital microscopy. We found that HU308 (CB2 receptor agonist) reduced the number of adherent leukocytes in submucosal venules but did not restore muscular and mucosal villi FCD in endotoxemic mice. AM630 (CB2 receptor antagonist) maintained the level of adherent leukocytes induced by LPS but further reduced muscular and mucosal villi FCD. URB597 (FAAH inhibitor) and JZL184 (MAGL inhibitor) both reduced the number of adherent leukocytes in submucosal venules but did not restore the mucosal villi FCD. Using various compounds we have shown different mechanisms of activating CB2 receptors to reduce leukocyte endothelial interactions in order to prevent further inflammatory damage during sepsis.

Clinical and genetic heterogeneity in familial temporal lobe epilepsy.

PURPOSE: Familial forms of temporal lobe epilepsy have been described recently. A locus on ch 10q has been linked to partial epilepsy with auditory symptoms. We investigated the proportion of families segregating temporal lobe epilepsy (TLE) linked to ch 10q and sought to establish genotype-phenotype correlations. METHODS: We studied 15 unrelated families segregating TLE. A total of 153 individuals, including 79 patients, were analyzed in this study. Family members were genotyped for four polymorphic dinucleotide repeat markers: D10S185, D10S574, D10S577, and D10S192, which flank the 15-cM candidate interval on ch 10q. Two-point lod scores were calculated for each family separately. RESULTS: Fourteen of our families had ictal semiology of mesial temporal onset of seizures and magnetic resonance imaging (MRI) abnormalities in the mesial structures; only one family, with seven affected individuals, reported auditory symptoms and had normal MRIs. Pedigree analysis showed an autosomal dominant transmission with 0.75 penetrance. Only two families had informative lod scores. A large family, with 22 affected individuals segregating mesial TLE, had negative lod scores for all four markers genotyped. The lod scores were significantly negative (less than -2.00) up to 0.05 for D10S185, 0.10 for D10S574, 0.25 for D10S577, and 0.15 for D10S192. The single family with auditory symptoms had positive lod scores for all markers genotyped, with a Z max of 1.52 at 0.0 for D10S574. CONCLUSIONS: We identified two different clinical groups of families segregating TLE. Most families identified in this study had mesial TLE. Only one single family segregating lateral TLE was found. We significantly excluded linkage between familial mesial TLE and the locus on ch 10q. In addition, we showed evidence for linkage between one family with lateral TLE and markers on ch 10q. This is strong evidence for clinical and genetic heterogeneity among familial forms of TLE.

Enzyme-linked immunofiltration assay used in the screening of solid supports and immunoreagents for the development of an azinphos-methyl flow immunosensor.

Azinphos-methyl (AM), O,O-dimethyl S-[(4-oxo-1,2,3-benzotriazin-3(4H)-yl)methyl] phosphorodithioate, is a dithiophosphorous insecticide extensively used for the control of fruit culture pests. In this work the ELIFA system, initially developed and marketed to substitute conventional ELISA methods, was used for the screening of supports and immunoreagents in the development of a flow immunosensor to AM. The objective was to find the optimal antibody concentration, support quantity and enzymatic tracer concentration to develop a sensitive and reusable immunosensor. The influence of chitosan as protein stabilizing agent was also investigated. We observed that, on the basis of immunosorbent characterization, chitosan-modified silica with immobilized LIB-MFH14 monoclonal antibody (MAb) showed the best sensitivity, with a I(50) value of 6 nM AM. All of the immobilized MAbs either in alkylaminated or chitosan-modified silica showed I(50) values between 10 and 36 nM. Regarding the regeneration capability, the best desorption agent tested was 0.1 M glycine/HCl, pH 2.0, performing in most cases a 100% desorption after just one wash and maintaining the antibody activity even after 20 cycles of regeneration. The chitosan-modified silica seemed to be the best support for this purpose.

Reduced susceptibility to azithromycin and high percentages of penicillin and tetracycline resistance in Neisseria gonorrhoeae isolates from Manaus, Brazil, 1998.

BACKGROUND: The identification of Neisseria gonorrhoeae isolates resistant to antimicrobial agents currently recommended for the treatment of gonococcal infections continues to escalate globally. Thus, in some areas, resistance to fluoroquinolone drugs is commonplace; several reports document resistance to third-generation cephalosporins, and the sporadic isolation of spectinomycin-resistant isolates continues unabated. Gonococcal resistance to azithromycin, an antibiotic used for the primary treatment of gonococcal infections in some Latin American countries, also has been described. Because the prevalence of resistant isolates is insufficiently documented in many areas of Latin America, the efficacy of locally recommended therapies for gonococcal infections is often unknown. GOAL: To determine the antimicrobial susceptibility and strain types of N gonorrhoeae isolates collected in Manaus, Brazil. These data will establish antimicrobial susceptibility baseline data for the region as a reference point for future surveillance. STUDY DESIGN: Consecutive N gonorrhoeae isolates from urethral and endocervical specimens were collected and examined for identity, antimicrobial susceptibility, and strain type (plasmid content, tetM type, auxotype, and serovar). RESULTS: Most of the isolates (65/81; 85.2%) were resistant to tetracycline, penicillin, or both, with the majority (n = 62) carrying plasmid-mediated resistance to tetracycline (tetracycline-resistant N gonorrhoeae [TRNG]). All of the TRNG contained the Dutch-type tetM plasmid, and 18 were A/S class NR/IA-02. Penicillinase-producing N gonorrhoeae comprised 8.2% (7/81) of the isolates. Of these seven isolates, four also were TRNG, and two carried chromosomal resistance to tetracycline. The isolates were susceptible to ciprofloxacin, spectinomycin, and ceftriaxone. However, 23 isolates were characterized by reduced susceptibility to azithromycin (MIC, 0.25-0.5 microg/ml), and one isolate had reduced susceptibility to ciprofloxacin (MIC, 0.25 microg/ml). CONCLUSIONS: This study supports the continued use of third-generation cephalosporins, spectinomycin, and fluoroquinolone drugs for the primary treatment of gonococcal infections in Manaus. The occurrence of isolates with reduced susceptibility to azithromycin and ciprofloxacin underscores the importance of ongoing antimicrobial susceptibility monitoring to support decisions regarding appropriate drugs for the treatment of gonococcal infections.

Validation of national algorithms for the diagnosis of sexually transmitted diseases in Brazil: results from a multicentre study.

OBJECTIVE: To validate STD flow charts for the management of genital discharge and genital ulcer currently recommended by the National STD Control Programme in Brazil. METHODS: A study was conducted in five Brazilian STD clinics from January to June 1995. After an interview, a clinical examination was performed by a physician, who recorded a presumptive diagnosis, based on his/her clinical experience. This diagnosis was compared with a gold standard laboratory diagnosis in order to calculate sensitivity, specificity, and positive predictive value of the clinical diagnosis. The validity of the simulated national flow charts was assessed using the same method. RESULTS: A total of 607 men and 348 women participated in the study. Gonorrhoea was the aetiology most frequently detected in men with urethral discharge. The sensitivity of the clinical diagnosis was far lower than the sensitivity fo the national flow chart, using the syndromic approach, for both gonococcal and chlamydial urethritis. Adding a simple laboratory test (Gram stain) to the national flow chart increased the specificity and positive predictive value for gonorrhoea. Among the women with vaginal discharge, a cervical infection was detected in 17%, a vaginal infection in 74%, and mixed infection in 9%. The sensitivity of the diagnosis for cervical infection increased from 16% (clinical aetiological approach) to 54% (when adding a syndromic approach) and to 68% when adding a risk assessment, as in the national flow charts. The cure or improved rate of genital ulcers was 96% after 1 week. CONCLUSIONS: The results of the study will help to convince policy makers and those involved in training healthcare workers in Brazil of the public health advantages of the syndromic approach, as an essential part of STD/HIV control activities.