Immunohistochemical study of the P2X2 and P2X3 receptor subunits in rat and monkey sensory neurons and their central terminals

Of the cloned P2X receptor subunits, six are expressed in sensory neurons, suggesting that the native channels may be heteromultimers with diverse composition. It has been proposed that P2X2 and P2X3 form heteromultimers in sensory neurons. We further tested this hypothesis by examining the relationship of P2X2 and P2X3 immunocytochemically. In rat dorsal root and nodose ganglia, P2X2- and P2X3-immunoreactivity (-ir) were highly colocalized, although single-labeled cells were also present. In dorsal root ganglia (DRG), in some cases P2X2-ir appeared to be present in satellite cells. In dorsal horn of spinal cord, at low magnification the laminar localization of P2X2- and P2X3-ir overlapped, but at high magnification colocalization was rarely observed. In contrast, in the solitary tract and its nucleus (NTS), colocalization of P2X2- and P2X3-ir was seen at low and high magnification. These results suggest that the relationship of P2X2- and P2X3-ir is different in nodose and dorsal root ganglia and might reflect differences in the targeting of P2X receptors in different sensory neurons. In monkey, P2X2-ir was observed in DRG neurons and satellite cells and in dorsal horn of spinal cord. P2X3-ir was also seen in DRG neurons. However, the presence of P2X2-ir in NTS as well as the presence of P2X3-ir in spinal cord and NTS could not be established definitively. These results suggest species differences, although a more extensive study of primate sensory systems is necessary.     

A reappraisal of mouth-to-mouth ventilation during bystander-initiated cardiopulmonary resuscitation: a statement for Healthcare Professionals from the Ventilation Working Group of the Basic Life Support and Pediatric Life Support Subcommittees, American Heart Association

There is growing recognition that race and socioeconomic variables in health research demand greater attention. The investigators compared racial differences in health definition, health value, and health-promoting behavior of 62 pairs (N = 124) of Black and White college women matched on age, body mass index, and socioeconomic status. Both groups of women had similar definitions of health, valued health to the same extent, and reported similar levels of self-actualization, health responsibility, exercise, and stress management. Black women, relative to White women, practiced fewer nutrition behaviors and had less interpersonal support. Interventions to reduce health risk associated with nutrition practices of Black women are warranted and further research is needed to explore the influence of the social structure of educational institutions on interpersonal relationships and other health behaviors. When socioeconomic status is taken into consideration, Black and White college women demonstrated more commonalities in health behavior than differences.     

Identification and characterization of kinetically competent carbinolamine and alpha-iminoglutarate complexes in the glutamate dehydrogenase-catalyzed oxidation of L-glutamate using a multiwavelength transient state approach

A highly constrained and heavily overdetermined multiwavelength transient state kinetic approach has been used to study the oxidative deamination of L-glutamate catalyzed by beef liver glutamate dehydrogenase. Spectra generated using the known enzyme-reduced coenzyme-substrate spectrum served as models for deconvolution of kinetic scan data. Deconvolution of the multiwavelength time course array shows formation of three distinguishable intermediates in the reaction sequence, an ultrablue-shifted complex, an ultrared-shifted complex, and a blue-shifted complex. The ultrablue-shifted entity is identified as the enzyme-NADPH-alpha-iminoglutarate complex (ERI) and the ultrared as the enzyme-NADPH-alpha-carbinolamine complex (ERC). The blue-shifted complex is characterized as the E-NADPH-ketoglutarate species (ERK). The location of these species along the reaction coordinate has been determined and their kinetic competency in the reaction sequence has been established by fitting the concentration time courses of the components for both the alpha-deuterio- and the alpha-protio-L-glutamate reactions to the now highly constrained differential equations derived from a kinetic scheme involving the sequential formation of alpha-iminoglutarate, alpha-carbinolamine, and alpha-ketoglutarate-reduced coenzyme complexes, following the formation of two prehydride transfer complexes.     

Vascular endothelial growth factor (VEGF)-C synergizes with basic fibroblast growth factor and VEGF in the induction of angiogenesis in vitro and alters endothelial cell extracellular proteolytic activity

Vascular endothelial growth factor-C (VEGF-C) is a recently characterized member of the VEGF family of angiogenic polypeptides. We demonstrate here that VEGF-C is angiogenic in vitro when added to bovine aortic or lymphatic endothelial (BAE and BLE) cells but has little or no effect on bovine microvascular endothelial (BME) cells. As reported previously for VEGF, VEGF-C and basic fibroblast growth factor (bFGF) induced a synergistic in vitro angiogenic response in all three cells lines. Unexpectedly, VEGF and VEGF-C also synergized in the in vitro angiogenic response when assessed on BAE cells. Characterization of VEGF receptor (VEGFR) expression revealed that BME, BAE, and BLE cell lines express VEGFR-1 and -2, whereas of the three cell lines assessed, only BAE cells express VEGFR-3. We also demonstrate that VEGF-C increases plasminogen activator (PA) activity in the three bovine endothelial cell lines and that this is accompanied by a concomitant increase in PA inhibitor-1. Addition of alpha2-antiplasmin to BME cells co-treated with bFGF and VEGF-C partially inhibited collagen gel invasion. These results demonstrate, first, that by acting in concert with bFGF or VEGF, VEGF-C has a potent synergistic effect on the induction of angiogenesis in vitro and, second, that like VEGF and bFGF, VEGF-C is capable of altering endothelial cell extracellular proteolytic activity. These observations also highlight the notion of context, i.e., that the activity of an angiogenesis-regulating cytokine depends on the presence and concentration of other cytokines in the pericellular environment of the responding endothelial cell.     

Identification of 3-hydroxypalmitic acid methyl ester as a novel autoregulator controlling virulence in Ralstonia solanacearum

A specific polyclonal antibody against the lipid peroxide (LOOH)-modified rabbit serum albumin (RSA) was generated in rabbits. The antibody selectively recognized the modified protein in a concentration-dependent manner and did not cross react with aldehyde-modified proteins or proteins directly oxidized with the free radical generator 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH). Oxidized low-density lipoprotein (Ox-LDL), but not native LDL, was also recognized by the antibody in a concentration-dependent manner. The antibody also cross reacted with several other proteins modified by LOOH suggesting that the antibody is directed towards a common epitope and not towards the protein sequence. Western blot analysis of normal human plasma showed that at least three different proteins are recognized by the antibody. RAW cells, preincubated with LOOH, were immunostained with the antibody and the antigenic epitopes were present intracellularly, while controls lacking in the primary antibodies failed to show immunoreactivity. Atherosclerotic arteries from cholesterol-fed monkeys and human atherosclerotic lesions were also immunostained by the antibody. The immunoreactivity was co-localized in areas rich in foam cell macrophages. These results suggest that LOOH-modified proteins present an unique antigenic epitope that may represent a primary product of interaction of LOOH with proteins.     

Helicobacter pylori infection and socioeconomic factors among adult Danes

The aim of this study was to examine the relationship between housing conditions, educational level, occupational factors, and serologically diagnosed acute and chronic Helicobacter pylori infection. IgG, IgM, and IgA antibodies against Helicobacter pylori were measured by ELISA techniques in sera obtained from a random sample of 3589 Danes participating in a population study. Poor social status (odds ratio 2.2 [1.7-3.0]), short duration of schooling (odds ratio 2.0 [1.3-2.5]), lack of vocational training/education (odds ratio 1.4 [1.2-1.7]), unskilled work (odds ratio 1.7 [1.2-2.5]), and high work-related energy expenditure (odds ratio 1.4 [1.1-1.9]) increased the likelihood of chronic Helicobacter pylori infection. Chronic Helicobacter pylori infection was frequently found in people with a history of living abroad. Increased levels solely of IgM antibodies to H. pylori, interpreted as a sign of acute infection, were found more often in people who were divorced (odds ratio 2.3 [1.2-4.4]) or unmarried (odds ratio 2.0 [1.1-3.8]) and in people who worked long hours (odds ratio 2.0 [1.1-4.0]). In conclusion, educational and occupational factors relate to the likelihood of chronic Helicobacter pylori infection in adults. The rate of acute Helicobacter pylori infection is probably increased in single adults.     

Musculoskeletal health surveillance. Eight simple, self-administered tests to add to symptom registration: a preliminary report

STUDY DESIGN: A population-based evaluation including cross-sectional comparisons of eight simple function tests (mobility and strength) and answers on questions concerning lifestyle factors, work environment, and health. The study population was randomly selected among people 35-45 years old. OBJECTIVES: To construct a simple, self-administered surveillance system to test musculoskeletal function to be mailed to a sample population along with a questionnaire. SUMMARY OF BACKGROUND DATA: A surveillance system should be easy to use, detect treatable symptoms at an early stage, and detect change of function or symptoms. METHOD: Questionnaires, which also contained instructions in performing the eight tests of musculoskeletal function, were sent to a randomly selected sample population. The questionnaire covered type and level of musculoskeletal problems, and lifestyle and occupational factors. RESULTS: The questionnaire and the self-administered test were completed by 834 people. The test movements could be performed and the questions answered by the study population. The tests discriminated between those with and without moderate or severe problems. Performance of physically heavy work did not significantly correlate with the test results. The results of the physical function test discriminated significantly between those with different levels of fear of movement (odds ratio, 2.2) and the degree of current somatic distress (odds ratio, 5.9). CONCLUSION: The test of physical function could be performed and the questionnaire completed by this randomly selected cohort, and the results discriminated significantly between groups.     

Ethanol inhibits single-unit responses in the nucleus accumbens evoked by stimulation of the basolateral nucleus of the amygdala

We studied the actions of intoxicating doses of ethanol on excitatory inputs from the basolateral nucleus of the amygdala, a major afferent system projecting to the nucleus accumbens (NAcc). In view of the hypothesized role of opioid receptors on the effects of ethanol on NAcc physiology, we also explored whether naloxone modulates ethanol-induced suppression of NAcc excitability in halothane anesthetized and freely moving unanesthetized rats. Intraperitoneal administration of ethanol (1.2-1.4 g/kg) markedly suppressed a subgroup of amygdala-activated NAcc neurons. The ethanol-induced reduction in amygdala-activated NAcc neurons was not reversed by naloxone (5.0 mg/kg, intraperitoneally). Moreover, naloxone had no effect on the suppressive effects of ethanol on NAcc spontaneous activity in either halothane-anesthetized or unanesthetized freely moving preparations. These findings suggest that opiate mechanisms either are not participating or are not solely responsible for the inhibitory effects of acute intoxicating doses of ethanol on NAcc physiology.