Oligoclonality of serum immunoglobulin G antibody responses to Streptococcus pneumoniae capsular polysaccharide serotypes 6B, 14, and 23F

Serum antibodies (Abs) specific for the capsular polysaccharides of Streptococcus pneumoniae provide protection against invasive pneumococcal disease. Previous studies indicate that Abs to pneumococcal polysaccharide (PPS) serotypes 1 and 6B have limited clonal diversity. To determine if restricted diversity was a feature common to other PPS specificities, we examined the light (L)-chain expression and isoelectric heterogeneity of type 6B, 14, and 23F Abs elicited in 15 adults following PPS vaccination. At the population level, both PPS-6B and PPS-14 Abs expressed kappa and lambda chains, although 6B Abs more frequently expressed lambda chains lambda and 14 Abs more frequently expressed kappa chains. In individual sera, Abs were generally skewed towards either kappa or lambda expression. 23F-specific Abs had predominantly kappa chains. Isoelectric focusing analyses showed that sera contained one or at most a few immunoglobulin G Ab spectrotypes to all three respective capsular serotypes, a result indicative of oligoclonality. A sequence analysis of a purified PPS-14-specific Ab having a single spectrotype gave uniform amino-terminal sequences for both the heavy chain (V(H)III subgroup) and the L chain (kappaIII-A27 V region). From these results we conclude that within individual adults, serum Ab responses to PPS serotypes 6B, 14, and 23F derive from a small number of dominant B-cell clones, and consequently variable-region expression is probably individually limited as well. Oligoclonality appears to be a general characteristic of human PPS-specific Ab repertoires, and we suggest that this property could lead to individual differences in Ab fine specificity and/or functional activity against encapsulated pneumococci.     

Dissecting the genetics of type 1 diabetes: relevance for familial clustering and differences in incidence

A combination of genetic and environmental factors is most likely the cause of Type 1 diabetes. Results from twin data, familial clustering of the disease and difference in incidence according to ethnicity infer the presence of specific disease genes. The genetic component of Type 1 diabetes cannot be classified according to a classical model of inheritance but is due to an interaction between different genes and environmental factors. The major genes are within the HLA region that are responsible for 40% of the genetic susceptibility, although other genes are important (non-HLA genes). To date, more than 10 specific loci have been localized on different chromosomes. The gene involved has been characterized only for two of such loci, IDDM1 and IDDM2, while in the other cases the presence of some susceptibility genes can be envisaged and their identification represents the goal of genetic research in coming years. Fine mapping of the loci will certainly increase our understanding of the genetics of Type 1 diabetes; the limitation in detecting some of the remaining genes by linkage studies can be overcome by association studies. That is possible via the collection of a large number of affected families (over 1000) in homogeneous populations.     

SAP90 binds and clusters kainate receptors causing incomplete desensitization

The mechanism of kainate receptor targeting and clustering is still unresolved. Here, we demonstrate that members of the SAP90/PSD-95 family colocalize and associate with kainate receptors. SAP90 and SAP102 coimmunoprecipitate with both KA2 and GluR6, but only SAP97 coimmunoprecipitates with GluR6. Similar to NMDA receptors, GluR6 clustering is mediated by the interaction of its C-terminal amino acid sequence, ETMA, with the PDZ1 domain of SAP90. In contrast, the KA2 C-terminal region binds to, and is clustered by, the SH3 and GK domains of SAP90. Finally, we show that SAP90 coexpressed with GluR6 or GluR6/KA2 receptors alters receptor function by reducing desensitization. These studies suggest that the organization and electrophysiological properties of synaptic kainate receptors are modified by association with members of the SAP90/PSD-95 family.     

Protein phosphorylation and taurine biosynthesis in vivo and in vitro

Taurine is known to be involved in many important physiological functions. Here we report that both in vivo and in vitro the taurine-synthesizing enzyme in the brain, namely cysteine sulfinic acid decarboxylase (CSAD), is activated when phosphorylated and inhibited when dephosphorylated. Furthermore, protein kinase C and protein phosphatase 2C have been identified as the enzymes responsible for phosphorylation and dephosphorylation of CSAD, respectively. In addition, the effect of neuronal depolarization on CSAD activity and 32P incorporation into CSAD in neuronal cultures is also included. A model to link neuronal excitation and CSAD activation by a Ca2+-dependent protein kinase is proposed.     

Corporeal veno-occlusive dysfunction: predominantly intracavernous muscular pathology

PURPOSE: We investigated whether a relationship exists between the flow to maintain an erection obtained at cavernosometry and the alteration of intracavernous structures in impotent patients with corporeal veno-occlusive dysfunction and normal arterial inflow. MATERIALS AND METHODS: Computerized histomorphometric analysis of smooth muscle and elastic fibers, and endothelial cells was compared to the flow necessary to maintain erection after intracavernous vasoactive drug injection in 18 patients with corporeal veno-occlusive dysfunction. RESULTS: A significant correlation between percentage of smooth muscle fibers and flow to maintain erection was observed, while no correlation was noted with elastic fibers and endothelial cells. CONCLUSIONS: Corporeal veno-occlusive dysfunction seems to be due mainly to smooth muscular alterations. According to this observation treatment of impotent patients with this abnormality should not be restricted to the penile veins but should also include the intracavernous structures, predominantly the muscular component.     

The effect of dietary oil containing (n-3) fatty acids on the fatty acid, physicochemical, and organoleptic characteristics of pig meat and fat

An investigation was made to alter the fatty acid composition of pork and a pork product in line with human dietary advice while not adversely affecting factors controlling consumer acceptability. Pigs (n = 150) were assigned to three dietary treatments with 25 intact male-female pairs per treatment. Diet A (control) contained 3% of a 4:1 (wt/ wt) tallow-soybean oil mixture. Diets B and C contained 2% rapeseed oil plus 1% fish oil. Diets A, B, and C were supplemented with 100, 100, and 250 mg of all-rac-alpha-tocopheryl acetate/kg of diet, respectively. Pigs were given ad libitum access to feed from 52 kg live weight until 95 kg (slaughter). Sausages were prepared from the resulting cuts. Tissues of pigs were evaluated in terms of fat firmness, color, fatty acid composition, and contents of alpha-tocopherol and thiobarbituric acid-reactive substances (TBARS). Organoleptic characteristics of chops and sausages were evaluated by a trained taste panel. Pigs fed Diets B and C had improved feed conversion ratios (P < .05) and ADG compared with control pigs. The levels of n-3 (omega-3) polyunsaturates were significantly increased in the tissues and sausage from pigs fed Diets B and C with associated alterations in n-6 to n-3 fatty acid ratios that accorded with contemporary human dietary recommendations. Levels of alpha-tocopherol and TBARS were significantly altered in the tissues. There were no appreciable differences between treatments in carcass characteristics, including color. The overall organoleptic acceptability of chops and sausages was not different between the treatments.     

Evaluation of client preference for function-based treatment packages

Functional communication training (FCT) and noncontingent reinforcement (NCR) are commonly prescribed treatments that are based on the results of a functional analysis. Both treatments involve delivery of the reinforcer that is responsible for the maintenance of destructive behavior. One major difference between the two treatment procedures is that client responding determines reinforcement delivery with FCT (e.g., reinforcement of communication is delivered on a fixed-ratio 1 schedule) but not with NCR (e.g., reinforcement is delivered on a fixed-time 30-s schedule). In the current investigation, FCT and NCR were equally effective in reducing 2 participants' destructive behavior that was sensitive to attention as reinforcement. After the treatment analysis, the participants' relative preference for each treatment was evaluated using a modified concurrent-chains procedure. Both participants demonstrated a preference for the FCT procedure. The results are discussed in terms of treatment efficacy and preference for control over when reinforcement is delivered. In addition, a method is demonstrated in which clients with developmental disabilities can participate in selecting treatments that are designed to reduce their destructive behavior.     

Minimal bystander activation of CD8 T cells during the virus-induced polyclonal T cell response

Acute infections with viruses such as lymphocytic choriomeningitis virus (LCMV) are associated with a massive polyclonal T cell response, but the specificities of only a small percentage of these activated T cells are known. To determine if bystander stimulation of T cells not specific to the virus plays a role in this T cell response, we examined two different systems, HY-specific T cell receptor (TCR)-transgenic mice, which have a restricted TCR repertoire, and LCMV-carrier mice, which are tolerant to LCMV. LCMV infection of HY-transgenic C57BL/6 mice induced antiviral CTLs that lysed target cells coated with two of the three immunodominant epitopes previously defined for LCMV (glycoprotein 33 and nucleoprotein 397). Although LCMV-induced cytotoxic T lymphocytes (CTLs) from C57BL/6 mice could lyse uninfected H-2(k) and H-2(d) allogeneic targets, LCMV-induced CTLs from HY mice lysed only the H-2(k)-expressing cells. The HY mice generated both anti-H-2(k) and anti-H-2(d) CTL in mixed leukocyte reactions, providing evidence that the generation of allospecific CTLs during acute LCMV infection is antigen specific. During the LCMV infection there was blastogenesis of the CD8+ T cell population, but the HY-specific T cells (as determined by expression of the TCR-alpha chain) remained small in size. To examine the potential for bystander stimulation under conditions of a very strong CTL response, T cell chimeras were made between normal and HY mice. Even in the context of a normal virus-induced CTL response, no stimulation of HY-specific T cells was observed, and HY-specific cells were diluted in number by day 9 after infection. In LCMV-carrier mice in which donor and host T cells could be distinguished by Thy1 allotypic markers, adoptive transfer of LCMV-immune T cells into LCMV-carrier mice, whose T cells were tolerant to LCMV, resulted in activation and proliferation of donor CD8 cells, but little or no activation of host CD8 cells. These results support the hypothesis that the massive polyclonal CTL response to LCMV infection is virus-specific and that bystander activation of non-virus-specific T cells is not a significant component of this response.     

beta-Lactamases responsible for resistance to expanded-spectrum cephalosporins in Klebsiella pneumoniae, Escherichia coli, and Proteus mirabilis isolates recovered in South Africa

Although resistance to the expanded-spectrum cephalosporins among members of the family Enterobacteriaceae lacking inducible beta-lactamases occurs virtually worldwide, little is known about this problem among isolates recovered in South Africa. Isolates of Klebsiella pneumoniae, Escherichia coli, and Proteus mirabilis resistant to expanded-spectrum cephalosporins recovered from patients in various parts of South Africa over a 3-month period were investigated for extended-spectrum beta-lactamase production. Antibiotic susceptibility was determined by standard disk diffusion and agar dilution procedures. Production of extended-spectrum beta-lactamases was evaluated by using the double-disk test, and the beta-lactamases were characterized by spectrophotometric hydrolysis assays and an isoelectric focusing overlay technique which simultaneously determined isoelectric points and general substrate or inhibitor characteristics. DNA amplification and sequencing were performed to confirm the identities of these enzymes. The P. mirabilis and E. coli isolates were found to produce TEM-26-type, SHV-2, and SHV-5 extended-spectrum beta-lactamases. An AmpC-related enzyme which had a pI of 8.0 and which conferred resistance to cefoxitin as well as the expanded-spectrum cephalosporins was found in a strain of K. pneumoniae. This is the first study which has identified organisms producing different extended-spectrum beta-lactamases from South Africa and the first report describing strains of P. mirabilis producing a TEM-26-type enzyme. The variety of extended-spectrum beta-lactamases found among members of the family Enterobacteriaceae isolated from major medical centers in South Africa is troubling and adds to the growing list of countries where these enzymes pose a serious problem for antimicrobial therapy.