Effects of fibinolysis on neurite growth from dorsal root ganglia cultured in two- and three-dimensional fibrin gels

The mechanism of neurite penetration of three-dimensional fibrin matrices was investigated by culturing embryonic chick dorsal root ganglia (DRGs) within fibrin gels, upon fibrin gels, and upon laminin. The length of neurites within three-dimensional matrices of fibrin was decreased in a concentration-dependent manner by agents that inhibited plasmin, e.g. aprotinin, or that inhibited plasminogen activation, e.g., epsilon-aminocaproic acid (EACA), or plasminogen antiserum. In contrast, such agents increased the length of neurites growing out from DRGs cultured upon two-dimensional substrates of fibrin and had no effect on the length of neurites growing out from DRGs cultured upon laminin. Visualization of neurites within three-dimensional fibrin matrices demonstrated that the distance between fibrin strands was much smaller than the diameter of neurites. All these data were consistent with the hypothesis that fibrinolysis localized to the region of the neurite tip is an important mechanism for neurite penetration of a physical barrier of fibrin strands arranged in a three-dimensional matrix.     

Dose-related effects of Ampligen (poly(I).poly(C12U)), a mismatched double-stranded RNA, in a bleomycin-mouse model of pulmonary fibrosis

The antifibrotic effect of the mismatched double-stranded RNA, Ampligen (poly(I).poly(C12U)), was evaluated in a bleomycin-mouse model of pulmonary fibrosis. Mice received a single intratracheal dose of bleomycin (0.125 U/mouse) or saline (50 microL) at the beginning of the experiment, followed by 5 or 6 intraperitoneal injections of Ampligen (1.0, 5.0, 10.0, 15.0, or 25.0 mg/kg) or saline at regular intervals for 2 weeks. Ampligen did not produce increased mortality or weight loss by itself. However, it produced varying degrees of mortality in combination with bleomycin. Five injections of 10 mg/kg Ampligen or three injections of 25 mg/kg Ampligen plus three injections of 10 mg/kg Ampligen in combination with bleomycin .produced significant reductions in lung collagen accumulation as indicated by lung hydroxyproline content compared to the bleomycin control group. Animals receiving bleomycin plus Ampligen at all dosages had significantly reduced prolyl hydroxylase activity compared to the bleomycin control group. Lipid peroxidation and bronchoalveolar lavage fluid (BALF)-supernatant protein content for the groups receiving bleomycin plus Ampligen were not reduced compared to the bleomycin control group. In the BALF-supernatant, the activity of acid phosphatase, a lysosomal enzyme produced by neutrophils, monocytes, and macrophages, was significantly decreased in the group receiving bleomycin plus 10 mg/kg Ampligen. Also, selected BALF differential immune cell counts were reduced in some of the groups receiving bleomycin plus Ampligen, but not in a consistent or dose-dependent manner. The results of this study indicate that Ampligen can significantly reduce the bleomycin-induced increased collagen accumulation and may be therapeutically useful in the management of lung fibrosis in humans.     

Loss of heterozygosity and overexpression of the p53 gene in ovarian carcinoma

Eight anticonvulsant drugs-including clonazepam, diazepam and phenobarbital-were tested for their effects on GABA-stimulated chloride uptake in rat cerebral cortical microsacs (unfiltered synaptoneurosomes). "Mid" and "high" therapeutic concentrations were screened, and, if significant enhancement was found, full concentration-response tests were done. In the initial screens, enhancement of GABA-stimulated uptake was found only with phenobarbital, clonazepam and diazepam. In subsequent concentration-response tests, the effects of phenobarbital were found to occur throughout the range of normal, anticonvulsant concentrations, whereas the effects of clonazepam and diazepam were observed only above the concentrations normally used for the chronic control of seizures or anxiety. These data suggest that phenobarbital's anticonvulsant effects are mediated via the GABAA receptor complex, but that the low-dose effects of the benzodiazepines may be mediated via some other mechanism.     

A transmembrane form of annexin XII detected by site-directed spin labeling

Previous studies of the annexin family of Ca2+ binding proteins identified a soluble monomer in the absence of Ca2+ and a trimer adsorbed on the membrane surface in the presence of Ca2+. On the basis of site-directed spin-labeling studies of annexin XII at low pH, we now report a membrane-inserted form of the protein with a dramatically different structure. The data suggest that upon insertion a continuous transmembrane alpha-helix is reversibly formed from a helix-loop-helix motif in the solution structure. Other regions with similar membrane-insertion potential were identified in the amino acid sequence, and we propose that the corresponding helices come together to form an aqueous pore that mediates the ion channel activity reported for several annexins.     

Hepatic blood flow in horses during the recuperative period from maximal exercise

OBJECTIVE: To determine effects of walking or standing on hepatic blood flow of horses after brief, intense exercise. ANIMALS: 6 adult Thoroughbreds (4 mares, 2 geldings). PROCEDURE: Horses were preconditioned on a treadmill to establish uniform level of fitness. Once fit, treadmill speed causing each horse to exercise at 120% of maximal oxygen consumption was determined and used in simulated races at 14-day intervals. In a three-way crossover study, horses were exercised at a speed inducing 120% of maximal oxygen consumption until fatigued or for a maximum of 2 minutes. Three interventions were studied: resting on the treadmill (REST), exercised then standing on the treadmill for 30 minutes (MS), and exercised then walking at 2 m/s for 30 minutes (MW). At 60 seconds after completion of exercise, bromsulphalein (BSP) was infused IV, and blood samples were collected every 2 minutes for 30 minutes for analysis of BSP concentration. Hematocrit and plasma total solids concentration were measured. Pharmacokinetic parameters were derived, using nonlinear regression, and were compared, using Friedman's repeated measures analysis on ranks. RESULTS: Plasma BSP concentration was higher after exercise. Median hepatic blood flow (BSP clearance) decreased significantly from 23.8 (REST) to 20.7 (MS) and 18.7 (MW) ml/min/kg. Median steady-state volume of distribution of BSP decreased from 47.6 (REST) to 42.7 (MW) and 40.2 (MS) ml/kg. Differences among trials were not significant when horses walked or stood after exercise. CONCLUSIONS: Hepatic blood flow and pharmacokinetics of BSP are markedly altered immediately after exercise. Limiting movement of horses during this period did not affect hepatic blood flow.     

Increased use of mammography among Hispanic women: baseline results from the NCI Cooperative Group on Cancer Prevention in Hispanic Communities

BACKGROUND: The Healthy People 2000 report set the objective of increasing the percentage of women 40 or older who had ever received a mammogram and clinical breast examination to 80% by the year 2000. The report used a baseline of 36% for all American women and 20% for Hispanic women. The purpose of this study was to compare baseline estimates with data obtained in five Hispanic communities. METHODS: Common survey measures were administered in five studies participating in a National Cancer Institute Cooperative agreement. The surveys evaluated history of mammography in five Hispanic communities in the southwestern Unites States. RESULTS: Across the five communities, the rates of mammography use were significantly higher than the national baseline. Among women 40-49 years of age, 55% had completed mammography (95% confidence interval [CI] = 52%, 57%). Among women 50 years of age or older, 64% had received a mammogram (95% CI = 62%, 66%). Older women (above age 50) were significantly more likely to have completed the test than younger women (younger than age 50), and mammography was obtained less often among women who were uninsured and those who had lower levels of acculturation. CONCLUSIONS: We conclude that the rate of mammography use among Hispanic women has increased significantly over the last few years and that we are on track to reach the goal of 80% mammography compliance for Hispanic women 40 years and older by the year 2000.     

The value of baseline chest radiograph reports in the care of elderly patients in an emergency department

The authors used a before/after research design to determine the impact of written reports of baseline chest radiographs (CXRs) on the care of elderly patients during 48 emergency department visits. Internal medicine housestaff completed questionnaires that included inquiries about their treatment plans for the patients both before and after they reviewed reports of baseline CXR results. The investigators compared these answers to determine the proportion of patients for whom baseline CXR reports changed patient management. In addition, three faculty reviewers used the nominal group process, a consensus analysis approach, to measure treatment changes and to estimate benefit to the patients from the alterations in management. The housestaff indicated that written baseline CXR reports caused them to change treatment plans during only 2 (4%) of the 48 visits. In both cases, they provided aggressive treatment for congestive heart failure because of baseline CXR results. The faculty reviewers could find only one case in which baseline CXR findings might have changed their management plans. Moreover, they were not convinced that the treatment changes indicated by the housestaff actually benefitted the patients. These results suggest that written reports of baseline CXRs do not improve the care of acutely ill patients in an emergency department setting.     

Structure and function in rhodopsin: rhodopsin mutants with a neutral amino acid at E134 have a partially activated conformation in the dark state

The Glu-134-Arg-135 residues in rhodopsin, located near the cytoplasmic end of the C helix, are involved in G protein binding, or activation, or both. Furthermore, the charge-neutralizing mutation Glu-134 to Gln-134 produces hyperactivity in the activated state and produces constitutive activity in opsin. The Glu/Asp-Arg charge pair is highly conserved in equivalent positions in other G protein-coupled receptors. To investigate the structural consequences of charge-neutralizing mutations at Glu-134 and Arg-135 in rhodopsin, single spin-labeled side chains were introduced at sites in the cytoplasmic domains of helices C (140), E (227), F (250), or G (316) to serve as "molecular sensors" of the local helix bundle conformation. In each of the spin-labeled rhodopsins, a Gln substitution was introduced at either Glu-134 or Arg-135, and the electron paramagnetic resonance spectrum of the spin label was used to monitor the structural response of the helix bundle. The results indicate that a Gln substitution at Glu-134 induces a photoactivated conformation around helices C and G even in the dark state, an observation of potential relevance to the hyperactivity and constitutive activity of the mutant. In contrast, little change is induced in helix F, which has been shown to undergo a dominant motion upon photoactivation. This result implies that the multiple helix motions accompanying photoactivation are not strongly coupled and can be induced to take place independently. Gln substitution at Arg-135 produces only minor structural changes in the dark- or light-activated conformation, suggesting that this residue is not a determinant of structure in the regions investigated, although it may be functionally important.     

From ethnography to intervention: developing a breast cancer control program for Latinas

Latinas are less likely than Anglo women to have appropriate breast cancer screening for reasons that may include culturally based beliefs as well as socioeconomic factors. This study employed ethnographic methods to explore breast cancer-related knowledge, attitudes, and behaviors among Latinas, Anglo women, and physicians, tested the generalizability of the findings in a telephone survey of randomly selected women, and used the results to design a culturally sensitive breast cancer control intervention in Orange County, Calif. Respondents for the ethnographic interviews included 28 Salvadoran immigrants, 39 Mexican immigrants, 27 Chicanas (U.S.-born Latinas of Mexican heritage), 27 Anglo women, and 30 physicians selected through organization-based network sampling. Latinas had very different beliefs about risk factors for breast cancer and held more fatalistic attitudes about the disease. For example, they believed that trauma to the breast was among the most important risk factors. Results of a telephone survey of 1225 randomly selected women (269 U.S.-born Latinas, 425 Mexican immigrants, 109 other Latina immigrants, and 422 Anglo women) generally confirmed the dissimilar beliefs among Latinas and Anglo women. The findings influenced our decision to design and pilot-test a breast cancer control intervention based on Bandura's self-efficacy theory and Freire's empowerment pedagogy. The methodology and findings of this study have important implications for future cancer control research and interventions.     

Initial analyses of naltriben, a delta opioid receptor antagonist, as a putative medicine for treating cocaine abuse.

The authors explored naltriben's (NTB, a delta opioid antagonist) potential to be a pharmacotherapy for cocaine abuse. NTB (3–6 mg/kg) reduced rats' (Rattus norvegicus) intravenous, self-administration of cocaine, daily, across 5 days. NTB did not, however, interfere with rats' pressing for water. NTB (3 mg/kg) blocked cocaine's facilitation of pressing for brain stimulation, daily, for 5 days. As doses of NTB were explored, it was found that 10 mg/kg of NTB was lethal for about one third of the rats. Smaller doses (e.g., 3 mg/kg) gave some indications of toxicity as indexed by NTB's tendency to reduce pressing for brain stimulation by itself. NTB (3 mg/kg) induced a mild conditioned taste aversion but had no negative effects on rats' ability to learn and remember a sequence of mazes. Taken together, these results lead to the suggestion that opioidergic processes play an important role in mediating cocaine's reinforcement. Although NTB may not be the ideal opioid antagonist for treating people, it has many positive properties supporting further investigation of opioid antagonists as agents for treating cocaine addiction. (PsycINFO Database Record (c) 2010 APA, all rights reserved)