Kinetics and quantitation of eosinophil and neutrophil recruitment to allergic lung inflammation in a brown Norway rat model

We quantitated neutrophil and eosinophil migration into lung parenchyma using specific peroxidase enzyme assays, and into the bronchoalveolar compartment by bronchoalveolar lavage (BALF), in sensitized brown Norway (BN), Fischer, and Lewis rats and also assessed the lungs by histopathology. Fourteen days after sensitization with ovalbumin (OA in alum [given subcutaneously] and OA with Bordetella pertussis [given intraperitoneally]), rats were challenged with an OA aerosol for 1 h. In BN rats, there was marked perivascular and peribronchial edema, focal hemorrhages, and increase in lung wet weight and BALF protein content, accompanied by neutrophilic infiltration at 3-14 h postchallenge. Few eosinophils were seen at 14 h in lung tissue or in BALF. Neutrophils peaked at 24 h in parenchyma ([94 +/- 7] x 10[6]) and in BALF ([2.7 +/- 0.4] x 10[6]) and declined rapidly thereafter. Marked eosinophil infiltration into parenchyma was apparent by 24 h. Eosinophil accumulation peaked at 48 h in parenchyma ([127 +/- 18] x 10[6]) and at 72 h in BALF ([10 +/- 2.4] x 10[6]), comprising up to 85% of lavage cells at this time. Lung eosinophilia persisted for at least 6 d with only a slow decline or clearance, not approximating baseline until day 13 after challenge. Histopathology showed peribronchial and interstitial eosinophilic pneumonia, most severe on day 3. In contrast to the BN rats, essentially no pulmonary inflammation was observed in Lewis and Fischer rats. This model in the BN rat, and the specific peroxidase assays for quantitating tissue eosinophils and neutrophils, should be useful for investigating the regulation of allergen-induced eosinophil and neutrophil migration into and clearance from the lung.     

Kinematic analysis of patients with spinal muscular atrophy during spontaneous breathing and mechanical ventilation

Dinoseb is a herbicide known to inhibit photosystem II electron transfer like DCMU, triazine and phenolic-type herbicides. The mutant Din7 of the cyanobacterium Synechocystis sp. PCC 6803, selected for resistance to dinoseb, and the mutant Ins2, constructed by the insertion of the kanamycin resistance cassette into the drgA gene, were cross-resistant to other nitrophenolic herbicides (DNOC, 2,4-dinitrophenol) and to the cell inhibitor metronidazole but not to the photosystem II inhibitors DCMU or ioxynil. The Din7 mutant had the same characteristics of photosystem II inhibition by dinoseb as the wild type. This result suggested the existence of another site for dinoseb inhibition. The wild type cells modified dinoseb to a non-toxic product that gave an absorption spectrum similar to that of dithionite treated dinoseb containing reduced nitro groups. In contrast, the Din7 mutant did not modify dinoseb. These phenomena were controlled by the drgA gene encoding a protein which showed similarity to several enzymes having nitroreductase activity. The addition of superoxide dismutase to the medium relieved the toxic effect of dinoseb in wild type cells but not in Din7. It is proposed that in wild type cells of Synechocystis sp. PCC 6803 the DrgA protein is involved in detoxification of dinoseb via the reduction of the nitro group(s) and this process is accompanied by the formation of toxic superoxide anions. Mutations blocking the activity of the DrgA protein lead to the development of resistance to nitrophenolic herbicides and metronidazole.     

Phenotypic expression of a mannose-sensitive hemagglutinin by a Vibrio cholerae O1 E1Tor strain and evaluation of its role in intestinal adherence and colonization

A Vibrio cholerae O1 strain (1150) of the EIT or biotype and Ogawa serotype with haemagglutination (HA) activity was subjected to TnphoA mutagenesis. Out of several mutants isolated, one HA- and another HA+ mutant were further characterised. The HA- mutant showed about 50% reduction in its intestinal adherence capacity in vitro and about 9-fold decrease of its colonisation ability in vivo, as compared to the wild-type strain. Subsequent studies showed that the HA activity of strain 1150 was mediated by a mannose-sensitive haemagglutinin (MSHA). Thus, the phenotypic expression of MSHA appears to be partly responsible for the intestinal adherence and colonisation properties of strain 1150.     

Partnering with physicians to achieve quality improvement

BACKGROUND: The Maine Medical Assessment Foundation (MMAF) has successfully involved hundreds of Maine physicians in study groups to analyze data on small-area variation and assess physician decision-making patterns. In 1991 the MMAF model was replicated across a tri-state area (Maine, New Hampshire, Vermont) in an effort called the Outcomes Dissemination Project, which is funded by a five-year grant from the U.S. Agency for Health Care Policy and Research. THE OUTCOMES DISSEMINATION PROJECT: Five specialty study groups, each meeting three times a year, examine local and national utilization data, examine guidelines and research findings, participate in outcomes studies and patient education, and disseminate their findings through specialty society presentations and other feedback efforts. The MMAF study group process is based on the beliefs that medicine is a subculture with a complex set of professional values, beliefs, socialization processes, and norms, and that quality improvement efforts work best when they are nonpunitive and educational. ISSUES IN OBTAINING PHYSICIAN INVOLVEMENT: (1) Physicians are willing to change their practices if they are brought into a culturally appropriate improvement program. (2) Related specialties (for example, internists and family practitioners) can often work together effectively on issues of common interest. (3) Involving respected clinical leaders has helped establish the legitimacy of MMAF methods among physicians. (4) Area- and physician-specific data are not made public, so as to build a sense of confidentiality among participants. CONCLUSIONS: The project continues to function as a powerful education process and serves as a model for replication elsewhere.     

The Drosophila G-protein-coupled receptor kinase homologue Gprk2 is required for egg morphogenesis

G protein signaling is a widely utilized form of extracellular communication that is mediated by a family of serpentine receptors containing seven transmembrane domains. In sensory neurons, cardiac muscle and other tissues, G protein-coupled receptors are desensitized through phosphorylation by a family of kinases, the G protein-coupled receptor kinases (GRKs). Desensitization allows a cell to decrease its response to a given signal, in the continued presence of that signal. We have identified a Drosophila mutant, gprk2(6936) that disrupts expression of a putative member of the GRK family, the G protein-coupled receptor kinase 2 gene (Gprk2). This mutation affects Gprk2 gene expression in the ovaries and renders mutant females sterile. The mutant eggs contain defects in several anterior eggshell structures that are produced by specific subsets of migratory follicle cells. In addition, rare eggs that become fertilized display gross defects in embryogenesis. These observations suggest that developmental signals transduced by G protein-coupled receptors are regulated by receptor phosphorylation. Based on the known functions of G protein-coupled receptor kinases, we speculate that receptor desensitization assists cells that are migrating or undergoing shape changes to respond rapidly to changing external signals.     

Metabolic efficiency during arm and leg exercise at the same relative intensities

This study was conducted to compare gross efficiency (GE), net efficiency (NE), work efficiency (WE), and delta efficiency (DE) between arm crank and cycle exercise at the same relative intensities. Eight college-aged males underwent two experimental trials presented in a randomized counterbalanced order. During each trial subjects performed three intermittent 7-min exercise bouts separated by 10-min rest intervals on an arm or semirecumbent leg ergometer. The power outputs for the three bouts of arm crank or cycle exercise corresponded to 50, 60, and 70% of the mode-specific VO2peak. GE, NE, and WE were determined as the ratio of Kcal.min-1 equivalent of power output to Kcal.min-1 of total energy expended, energy expended above rest and energy expended above unloaded exercise, respectively. DE was determined as the ratio of the increment of Kcal.min-1 of power output above the previous lower intensity to the increment of kcal.min-1 of total energy expended above the previous lower intensity. GE and NE did not differ between arm crank and cycle exercises. However, WE was lower (P < 0.05) during arm crank than cycle exercise at 50, 60, and 70% VO2peak. DE was also lower (P < 0.05) during arm crank than cycle exercise at delta 50-60 and at delta 60-70% VO2peak. It is concluded metabolic efficiency as determined by work and delta efficiency indices was lower during arm crank compared with cycle exercise at the same relative intensities. These findings add to the understanding of the difference in metabolic efficiency between upper and lower body exercise.