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Biomedical subjects

J E Repine

Publications and source records attributed to J E Repine.

At least 235 records · Page 13Linked to original sources
PubMed1977

Chemiluminescence by human alveolar macrophages: stimulation with heat-killed bacteria or phorobol myristate acetate.

Chemiluminescence of human alveolar macrophages (AM) was evaluated in vitro. Unstimulated AM generated chemiluminescence that remained constant during incubation. Addition of heat-killed Staphylococcus aureus 502A (HKB) or a chemical agent, phorbol myristate acetate, produced high rates of chemiluminescence that were significantly (P less than 0.05) increased over unstimulated AM. Phorbol myristate acetate-and HKB-stimulated increases in AM chemiluminescence were completely blocked by the enzyme superoxide dismutase. In comparison with unstimulated polymorphonuclear leukocytes, unstimulated AM had significantly (P less than 0.005) greater levels of chemiluminescence. However, after stimulation by phorbol myristate acetate or HKB, AM showed less chemiluminescence than similarly treated polymorphonuclear leukocytes.

Humans↗
PubMed1977

Influence of a deficiency of the second component of complement on the bactericidal activity of neutrophils in vitro.

Serum from three patients with a complete, selective deficiency of the second component of complement (C2) did not promote optimal killing of Staphylococcus aureus, 502A by neutrophilic polymorphonuclear leukocytes (PMN) in vitro. The addition of C2 reagent or the presence of heat-stable opsonin in the C2-deficient serum corrected the defective killing of S. aureus that was observed with patient or control PMN. PMN from the patients or control subjects killed bacteria with equal efficiency under conditions of optimal opsonization (normal pooled serum). However, twice-washed control PMN were better than patient PMN in killing S. aureus under circumstances of suboptimal opsonization (C2-deficient serum, heated C2-deficient serum, heated normal pooled serum, or no replacement of serum). The latter finding was due to residual C2 on the surface of twice-washed control cells. As repeated washing of control PMN progressively removed cell-associated C2, the staphylocidal effectiveness of the control RMN decreased to the level of patient PMN. In contrast to the findings with S. aureus, triply-washed PMN from patients or controls killed normal numbers of Escherichia coli, ON2, in C2-deficient serum.

Animals↗
PubMed1977

Quantitative measurement of the bactericidal capability of neutrophils from patients and carriers of chronic granulomatous disease.

Previous studies examined the ability of polymorphonuclear leukocytes (PMN) from patients and carriers of chronic granulomatous disease (CGD) to kill small numbers of bacteria in vitro. The purpose of the present study was to accurately measure the number of microorganisms killed by CGD PMN when the initial number of bacteria per neutrophil was increased. Bactericidal capacities of PMN from seven patients and five proven carriers of CGD were examined with five test ratios ranging from 1.25 to 100 bacteria/neutrophil. CGD PMN killed a limited number of catalase-positive S. aureus but the number of bacteria killed increased progressively as the initial test ratio was advanced. At the highest ratio (100:1) patient PMN killed a mean of nine S. aureus per cell (normal 48 +/- 13.2 SD). PMN from patients with CGD killed increasing numbers of catalase-negative S. faecalis with normal efficiency, reaching a mean of 14 S. faecalis killed per cell at 100:1. Individual variation in the ability of CGD PMN to kill S. Aureus was apparent but the degree of PMN bactericidal effectiveness did not correlate with the patient's clinical course. PMN from asymptomatic carriers of proven sex-linked CGD also had a wide spectrum of staphylocidal activity which ranged from normal to levels comparable to CGD patients.

Adult↗
PubMed1976

Primary leucocyte alkaline phosphatase deficiency in an adult with repeated infections.

An individual with repeated bacterial infections, eczema, hyperimmunoglobulin E, and a primary deficiency of leucocyte alkaline phosphatase is described. The Lap-deficient neutrophils from this patient had marginally deficient bactericidal activity particularly when challenged with high ratios of bacteria per neutrophil. Leucotactic, metabolic and morphologic features of the neutrophils from the patient were normal. Evidence is presented which contrasts this patient's condition with previously described primary or secondary deficiencies of LAP.

Adult↗
PubMed1976

Abnormal pattern of bactericidal activity of neutrophils deficient in granules, myeloperoxidase, and alkaline phosphatase.

The morphology, cytochemistry, metabolism, and bactericidal function of neutrophilic polymorphonuclear leukocytes (PMN) from a patient with subacute myelogenous leukemia were evaluated. The patient's mature PMN were deficient in granules and staining reactivity for myeloperoxidase (MPO) and alkaline phosphatase (LAP). These cells killed Staphylococcus aureus in an abnormal pattern when they were challenged with various increasing multiples of bacteria per neutrophil. At a low ratio of challenge (1.25 bacteria per neutrophil) the MPO-LAP-deficient PMN killed only 18 +/- 6 per cent (mean +/- 1 S.D.) (normal, 79 +/- 7) of the initial bacterial inoculum. As the PMN were challenged with higher ratios of bacteria per cell, the bactericidal effectiveness of the hypogranular PMN improved. At a 50:1 ratio the patient's cells killed within the normal range (28 +/- 10 per cent vs. normal of 48 +/- [mean +/- 1 S.D.]). Although rates of glucose oxidation and oxygen consumption by patient or control PMN stimulated with comparable ratios of heat-killed bacteria were the same, only minimal metabolic enhancement was produced in the MPO-LAP-deficient PMN by lower ratios. In contrast, higher ratios produced a marked increase in both of these metabolic activities indicating a major metabolic response to multiple ingestions by the patient's PMN. These observations may reflect the activation of compensatory microbicidal mechanisms available to the MPO-LAP-deficient PMN only when challenged by large multiples of bacteria.

Aged↗
PubMed1976

Quantitation of maximal bactericidal capability in human neutrophils.

The quantitative capacity of normal human neutrophils to ingest and kill increasing multiples of bacteria up to a level of saturation was evaluated. The in vitro system, a modification of classic bactericidal assay, employed carefully controlled ratios of bacteria per neutrophil between 1.25 and 400:1 to stress neutrophil function. Morphologic examination and measurement of bacterial survival by quantitative plate culturing indicated that at a challenge of about 100 Staphylococcus aureus per neutrophil bactericidal activity reached its peak with each average cell killing 48 bacteria. The efficiency of bactericidal function fell as the challenge was increased while ingestion capabilities kept pace with the numbers of bacteria presented to the neutrophils. Various key test parameters were evaluated from which evolved a system which provides a highly reproducible, statistically based, method for clinical assessment of subtle or partial neutrophil dysfunctions which may escape detection by the currently employed techniques of bactericidal assay.

Blood Bactericidal Activity↗
PubMed1976

Reversible neutrophil defect in patients with bacterial endocarditis.

The bactericidal capacities of neutrophilic polymorphonuclear neukocytes (PMN) from five untreated adults with bacterial endocarditis (BE), eight untreated patients with other forms of acute bacterial infection (ABI), and fourteen drug-free uniinfected normal subjects (NS) were compared with a new technique that uses five increasing bacteria-to-neutrophil ratios ranging from 1.25 to 100 Staphylococcus aureus per neutrophil. PMN from uninfected NS or untreated patients with ABI demonstrated a similar and reproducible ability to kill increasing numbers of S. aureus in 8 per cent normal serum. In contrast, the bactericidal activities of the PMN from untreated patients with BE were significantly depressed and the defect was more apparent at high ratios. Neutrophils from some of these individuals had decreased bactericidal action only at high ratios, indicating a quantitative type of neutrophil defect. There was no morphologic deficiency in the uptake of bacteria by BE neutrophils and comparable rates of glucose [-1-14C] oxidation were found in BE and control neutrophils stimulated with various ratios of heat-killed bacteria. Therefore, the observed abnormality appeared to be that of intracellular killing rather than of ingestion. The proportions of bacteria killed by the MN of untreated patients with BE improved after antibiotic treatment and became equal to those of NS or untreated ABI patients. This rapid return to normal bactericidal function by the PMN during treatment indicates that the prior deficiency was an acquired consequence of the infectious process of BE.

Adult↗
PubMed1975

Spectrum of function of neutrophils from carriers of sex-linked chronic granulomatous disease.

Bactericidal and metabolic activities were compared for polymorphonuclear leukocytes from normal controls, patients with sex-linked chronic granulomatous disease, five obligate carriers, and six potential carriers of CGD. Bacteria surviving at one hour were quantitated in a standardized assay which employed 1.25 Staphylococcus aureus per neutrophil. Heat-killed bacteria or a chemical agent, phorbol myristate acetate, were used to stimulate increases in utilization of oxygen, oxidation of [1--14C] glucose, and reduction of neotetrazolium chloride by PMN. The results demonstrate that PMN from the individual obligate carriers of CGD have a broad spectrum of functional capabilities. Neutrophils from one obligate carrier performed in the above in vitro tests and others on a par with normal control cells, whereas the PMN of others displayed deficiencies nearly as profound as those of the affected CGD patients. The observations parallel the broad range of phenotypic expression observed in heterozygotic carriers of other sex-linked recessive disorders as a result of random inactivation of the X chromosome. Although predictable from the current concept of random X inactivation, the spectrum has not been previously demonstrated for carriers of sex-linked recessive CGD and thus has important implications for the detection and counseling of carriers of CGD.

Blood Bactericidal Activity↗
PubMed1975

Deficiency of the second component of complement (C2) with chronic vasculitis.

A patient had complete deficiency of the second component of complement associated with chronic vasculitis and increased susceptibility to infection. We discuss here results of the complement profile, histocompatibility typing, and studies of the functional properties of patient plasma or serum in chemotaxis and opsonization in relation to the disease entity and host susceptibility to infection.

Adult↗
PubMed1975

The influence of phorbol myristate acetate on the metabolism of neutrophils from carriers of sex-linked chronic granulomatous disease.

The present investigation has compared the influences of phorbol myristate acetate (PMA) and heat-killed bacteria (HKB) on oxygen consumption and glucose oxidation by polymorphonuclear leukocytes (PMN) from carriers of sex-linked chronic granulomatous disease (CGD). PMA or HKB caused neutrophils from CGD carriers, considered as a group, to consume oxygen and oxidize glucose-1-14C at rates that were statistically distinguishable from rates of normal controls and affected CGD hemizygotes. PMA at a final concentration of 1.0 micrograms per milliliter wass more effective and reproducible than a ratio of 50 HKB: 1 PMN in discriminating the partial abnormality of carrier PMN from normal PMN. Moreover, a deficiency in glucose oxidation by the PMN of one individual carrier was detectable using PMA stimulation when no defect was apparent with HKB. Results of the present investigation confirm and extend previous observations which have demonstrated the similarity in responses of PMA-treated normal and CGD PMN to the reactions produced by particulates under similar conditions.

Alcohols↗
PubMed1974

Effects of phorbol myristate acetate on the metabolism and ultrastructure of neutrophils in chronic granulomatous disease.

Previous investigations have demonstrated that phorbol myristate acetate (PMA), the active principle of croton oil, stimulates alterations in normal polymorphonuclear leukocytes (PMN) that resemble closely the changes that develop in the cells after phagocytosis of bacteria. The present study has compared the effects of PMA and heat-killed bacteria on the oxygen uptake, glucose oxidation, nitroblue tetrazolium (NBT) reduction, and ultrastructure of normal neutrophils and PMN from six patients with chronic granulomatous disease (CGD). PMA stimulated oxygen consumption, hexose monophosphate shunt activity, and NBT reduction in normal cells but failed to produce similar effects in CGD neutrophils. However, PMA did induce formation of cytoplasmic vacuoles in the CGD cells similar to those observed in normal neutrophils. The results indicate that PMA is a useful nonparticulate agent for distinguishing between normal and CGD neutrophils and for studying basic mechanisms of phagocytosis in normal and abnormal PMN.

Alcohols↗
PubMedDate not supplied

Reversible pseudohyphal growth in haploid Saccharomyces cerevisiae is an aerobic process.

Pseudohyphal growth in Saccharomyces cerevisiae has been postulated to be an adaptation to foraging for nitrogen during nitrogen starvation. This process was described as a strictly diploid phenomenon which did not occur in haploid yeast cells and was under the genetic control of both the mating-type locus and a group of five genes, the BUD genes, regulating bud formation. We have also observed a dimorphic growth pattern in yeast growing on various nitrogen-limiting synthetic media. However, and in contrast to a previous report, we find that pseudohyphal growth is not precluded in haploid cells. We demonstrate that haploid pseudohyphal growth is strictly oxygen-dependent and is rapidly reversible, defining pseudohyphal growth as a reversible developmental pathway in yeast.

Haploidy↗
PubMedDate not supplied

Extracellular flush solution that contains blood, mannitol, albumin, and prostacyclin protects rat lungs from six hours of ischemia.

We have used an isolated rat lung model to compare the quality of preservation of different flush techniques with each other and with topical cooling alone. Lung injury was assessed by recording lung weights after reperfusion after 4 and 6 hours of ischemia. The flush solutions studied were intracellular (Collins-Sacks), traditional extracellular, extracellular with low potassium plus dextran, and extracellular containing blood, mannitol, albumin, and prostacyclin (Wallwork's solution). Flushing with Wallwork's solution before both 4 and 6 hours of ischemia gave superior protection from lung edema after reperfusion over all the other methods.

Albumins↗