Search PubMed⌕ Search

Biomedical subjects

T H Price

Publications and source records attributed to T H Price.

At least 55 records · Page 3Linked to original sources
PubMed1985

The role of neutrophil membrane glycoprotein GP-150 in neutrophil adherence to endothelium in vitro.

We have previously described two patients with a congenital defect in neutrophil function characterized by an inability to form pus. The patients' neutrophils lack a membrane glycoprotein of mol wt 150,000 daltons (GP-150) on analysis by SDS-PAGE. This glycoprotein is part of a membrane antigen complex recognized by the murine monoclonal antibody (MoAb) 60.3. Addition of MoAb 60.3 to normal neutrophils produces defects in chemotaxis and phagocytosis in vitro similar to those observed in the patients. Since neutrophil adherence to vascular endothelium is prerequisite to neutrophil emigration in vivo, we examined the interaction of the patients' neutrophils and normal neutrophils treated with MoAb 60.3 with cultured endothelium. Adherence was determined as the percentage of 51Cr-labeled purified peripheral blood neutrophils which remained adherent to plastic wells or endothelial monolayers after a 45-minute incubation at 37 degrees C. The percentage of neutrophils from patient 1 remaining adherent to uncoated, fibronectin-coated, or laminin-coated plastic was similar to that observed in normal neutrophils (55% to 84% adherence with normal neutrophils v 73% to 78% adherence with the patient's neutrophils and 63% to 82% adherence with MoAb 60.3-treated normal neutrophils). The adherence of the neutrophils from patient 1 and MoAb 60.3-treated normal neutrophils to human or bovine endothelium in serum-free medium was also not significantly different from that observed in normal neutrophils (less than 10% adherence with normal, MoAb 60.3-treated, and patient neutrophils). In medium containing 10% autologous or heterologous human plasma, however, the adherence of neutrophils from patient 1 or MoAb 60.3-treated normal neutrophils to endothelial monolayers was significantly reduced (35% +/- 7% of normal neutrophils in seven experiments). Although phorbol myristate acetate (PMA) (10 ng/mL) and calcium ionophore A23187 (10(-5) mol/L) markedly increased the adherence of normal neutrophils to endothelial monolayers in serum-free medium (40% to 85% adherence), neither agent increased the adherence of the neutrophils from patient 1 or normal neutrophils treated with MoAb 60.3 (less than 5% adherence). The adherence of PMA-activated neutrophils from patient 2 to endothelial monolayers was also markedly decreased when compared with that of normal neutrophils. Postsecretory cell-free supernatants from PMA-activated normal neutrophils failed to augment adherence of neutrophils from patient 1 (less than 5% adherence).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1984

Absence of monoclonal-antibody-defined protein complex in boy with abnormal leucocyte function.

An 8-year-old boy with severe recurrent bacterial infections was found to lack the cell-surface multimeric polypeptide complex defined by murine monoclonal antibody 60.3. This complex is expressed on the surface of all normal polymorphonuclear leucocytes (PMN), monocytes, and lymphocytes. Incubation of normal lymphocytes or PMN with antibody 60.3 produced functional abnormalities comparable with those observed in the patient's cells, suggesting that the polypeptide complex plays a significant part in several membrane-dependent leucocyte functions.

Antibodies, Monoclonal↗
PubMed1984

The use of Preston enrichment broth for the isolation of 'thermophilic' campylobacters from water.

The application of Preston enrichment broth to isolation of 'thermophilic' campylobacters from water has been investigated. Enrichment substantially increased the yield of such organisms. The optimum timing for subculturing the broth was after 48 h incubation. Despite the addition of aerotolerant supplement to the broth the number and variety of isolates was greater when the broth was incubated microaerically than when it was incubated in air.

Aerobiosis↗
PubMed1983

Chronic lymphocytosis with neutropenia: evidence for a novel, abnormal T-cell population associated with antibody-mediated neutrophil destruction.

A 67-year-old man with stable chronic neutropenia, lymphocytosis, multiple auto-antibodies, and recurring infections was studied in order to characterize the abnormal lymphocytes and the mechanism of neutropenia. One-half of the circulating mononuclear leukocytes were large granular lymphocytes that did not rosette with sheep erythrocytes and did not have surface immunoglobulin or receptors for C3 or IgG Fc. Virtually all of the mononuclear leukocytes, however, reacted with three monoclonal antibodies specific for mature T lymphocytes--OKT3, 3A1, and 12.1--the latter with a bimodal pattern on FACS analysis. The nonadherent, non-E-rosetting lymphocytes consisted of a homogeneous population of T cells with a novel phenotype: 3A1+, OKT3+, OKT8+, Ia+, 12.1+, OKT4-, 9.6-, 10.2-, Fc gamma receptor-. The abnormal phenotype of these cells suggested that they may have developed clonally. In spite of their OKT8+ phenotype, these lymphocytes were functionally abnormal, since they did not suppress B-cell immunoglobulin production in vitro. A humoral immune mechanism of neutropenia was indicated by increased levels of neutrophil-reactive IgG and in vivo kinetic studies with autologous neutrophils demonstrating shortened intravascular survival. The granulocyte turnover was normal, however, suggesting a blunted marrow response to the neutropenia. Corroborating the kinetic data, in vitro cultures of the patient's blood and marrow cells showed reduced numbers of granulocytic progenitors (CFU-C). Neither blood lymphocytes nor serum from the patient suppressed growth of allogeneic CFU-C nor did removal of OKT8+ cells from the marrow increase CFU-C expression. Thus, the disorder in this patient is characterized by proliferation of abnormal OKT8+ lymphocytes with impaired suppressor function. Our findings suggest that the neutropenia was due to anti-neutrophil autoantibodies that may have resulted from abnormal T-cell immunoregulation.

Aged↗
PubMed1983

Multiple plating from enrichment media as an aid to salmonella isolation.

Faeces samples enriched for salmonellas in selenite F broth incubated at 43 degrees C were subcultured to brilliant green MacConkey agar after 6 h. Four platings were made. As a control technique, each sample was enriched in selenite F incubated at 37 degrees C for 24 h and subcultured once to brilliant green MacConkey agar. Of the specimens plated at 6 h, out of a total of 25 positive samples, 6 required more than a single subculture to reveal the presence of salmonellas. Twenty-four positive isolations were made from the selenite broths incubated at 37 degrees C for 24 h and subcultured once only. All faecal samples positive by enrichment were also positive by direct plating. Nine samples seeded by 6 h produced only 1-2 salmonella colonies per plate. Convalescent specimens are, therefore, unlikely to be efficiently diagnosed by early subculture even if it is combined with multiple plating.River Taff samples, after pre-enrichment in buffered peptone water, were enriched in Rappaport's medium at 37 degrees C for 6 h. Subcultures were made to four plates of brilliant green MacConkey agar. A control subculture was made at 24 h to a single plate of brilliant green MacConkey. Although multiple subculture at 6 h improved salmonella isolation, the method was not as efficient as a single plating from the same enrichment culture at 24 h. There was no evidence that subculture at 6 h combined with multiple plating had a value either for faecal examination or for the isolation of salmonellas from sewage-polluted natural water.Samples of other natural waters submitted routinely by Environmental Health Officers were pre-enriched in buffered peptone water before subculture to Rappaport's medium. The enrichment cultures were incubated at 37 degrees C for 24 h and duplicate platings were made to brilliant green MacConkey agar. By double plating at 24 h, a gain of 16% in salmonella recovery was made. This compares favourably with the gain obtained by subculture from Rappaport after 24 and 48 h incubation. It also saves 24 h in examination time.

Bacteriological Techniques↗
PubMed1983

Salmonella isolation from reptilian faeces: a discussion of appropriate cultural techniques.

A short study of salmonella isolation from reptile faeces is described. The samples came from the reptile house at the Bristol Zoological Gardens. The wide range of salmonella serotypes present in the material, including representatives of all four subgenera, is noted. The important factors in the technique of isolation are discussed. These are choice of inoculum, enrichment medium, and plating medium, use of multiple subculture and serological isolation. If the number of samples available is scanty, an extended technique will probably yield more information than a simple method.

Alligators and Crocodiles↗
PubMed1983

Effect of stool storage at room temperature on salmonella isolation from faeces.

Stools, known to have contained salmonellas were cultured in three enrichment media on arrival at the laboratory. The three media were laboratory prepared selenite F, laboratory prepared Muller-Kauffmann tetrathionate and commercially obtained Oxoid Muller-Kauffmann tetrathionate (CM343). Where faecal samples were sufficiently large, they were stored at room temperature and re-examined using the same three enrichment broths. Whether fresh or stored stools were investigated, the laboratory prepared media were significantly more efficient than the commercial medium. In a straight comparison of the two laboratory prepared media, no significant difference in efficiency was evident with fresh stool samples. With stored specimens, however, Muller-Kauffmann tetrathionate was significantly better than selenite F. This finding might be relevant in examining samples delayed in the post.

Culture Media↗
PubMed1983

A comparison of two modifications of Rappaport's enrichment medium (R25 and RV) for the isolation of salmonellas from sewage polluted natural water.

The development of Rappaport's enrichment medium and elevated temperature incubation as methods of salmonella isolation is traced. The recent recording of a merger of the two techniques by means of Rappaport--Vassiliadis medium is noted (RV medium). In Cardiff, we have found an earlier modification of Rappaport's enrichment (R25) by Vassiliadis to be efficient in salmonella recovery from environment samples. The current study compares the two media using sewage polluted natural water as test material. Under the conditions of experiment, R25 was more successful in salmonella isolation than RV, although the later medium inhibited competitive organisms better. R25 is a convenient enrichment broth for routine use. In combination with pre-enrichment it allows the use of a loop for subculture rather than a pipette. This increases safe manipulation. It also produces a high proportion of positive isolations at the 24 h subculture time in contrast to other enrichment broths. For these reasons it forms an integral part of salmonella isolation methodology in our laboratory.

Agar↗
PubMed1983

Definition of a common leukocyte cell-surface antigen (Lp95-150) associated with diverse cell-mediated immune functions.

We have described a monoclonal antibody, designated 60.3, which reacts with a cell surface antigen expressed by most peripheral blood and bone marrow leukocytes. Immunoprecipitation showed at least three major components with relative m.w. of 95,000, 130,000, and 150,000 under reducing conditions. Antibody 60.3 inhibited several cell-mediated immune functions. The lytic activity of both cytotoxic T cells and natural killer cells was blocked in the presence of the antibody. The proliferative responses of T cells stimulated by soluble antigens, mitogens, or allogeneic cells were inhibited when antibody 60.3 was added at the initiation of culture, but not when added after 48 hr. Antibody 60.3 also blocked the migration of neutrophilic granulocytes. The antigen identified by antibody 60.3 thus appears to be involved in a membrane-dependent cell activation pathway that is common to diverse functional systems and is shared by both lymphoid and myeloid cells.

Animals↗
PubMed1982

Severe recurrent bacterial infections associated with defective adherence and chemotaxis in two patients with neutrophils deficient in a cell-associated glycoprotein.

We studied two patients with delayed umbilical cord detachment, recurrent bacterial infections, inability to form pus, rapidly progressive periodontitis, and persistent leukocytosis. The phagocytes of both patients were strikingly abnormal in their ability to adhere to surfaces. The adherence of polymorphonuclear leukocytes to endotoxin-coated glass coverslips, glass beads, or nylon wool was markedly reduced. Scanning electron microscopy of the few adherent polymorphonuclear leukocytes from both patients showed a failure to flatten and form fine pseudopods. In vivo polymorphonuclear leukocyte and monocyte chemotaxis assessed by skin window and skin chamber methods was dramatically impaired, and in vitro chemotaxis was severely depressed. Chemiluminescence of zymosan- but not phorbol-stimulated polymorphonuclear leukocytes was markedly reduced. Allogeneic polymorphonuclear leukocytes transfused into these patients functional normally, indicating that the defect is intrinsic to the cells and not a secondary phenomenon. A 180-kilodalton glycoprotein normally present in the particulate fraction of polymorphonuclear leukocytes was found to be completely absent in Patient 1 and present in low concentration in Patient 2. We postulate that the glycoprotein deficiency interferes with the migration of polymorphonuclear leukocytes from the bloodstream into the interstitial space and to the site of infection.

Bacterial Infections↗
PubMed1982

Influence of multiple plating from fluid media on salmonella isolation from animal feeding stuffs.

The influence of multiple plating of fluid cultures on salmonella isolation from animal feeding stuffs was examined. Four plating were made from broth culture after 24 h at 37 degrees C and four platings from selenite enrichment from 24 h at 43 degrees C. Selenite enrichment followed broth culture which was used as a pre-enrichment stage. Brilliant green MacConkey agar plates were employed for broth subculture and brilliant green MacConkey and desoxycholate citrate agars for selenite subculture. The eight brilliant green plates subcultured from broth and selenite were examined for salmonellas after incubation for 24 h at 37 degrees C. The four desoxycholate citrate agars after 24 h at 37 degrees C were used for motility enrichment. The food sample size was a single 100 g instead of 4 x 25 g cultured in an earlier study. This pooling of samples aimed at technical economy. Quadruple plating played an important part in salmonella isolation from 100 g specimens. The combination of multiple plating with motility enrichment was the most successful technique used.

Animal Feed↗
PubMed1982

A comparisoin of the R25 modification of Rappaport s enrichment medium with strontium chloride B for salmonella isolation from sewage polluted natural water.

The relation of salmonella isolation efficiency and the size of inoculum introduced from a buffered peptone water culture of sewage polluted water into strontium chloride B medium was investigated. Two separate studies were made, one using enrichment at 37 degrees C, the other at 43 degrees C. From these trials, two inocula suitable for efficient salmonella isolation were determined. Using this information, strontium chloride B medium was compared with modified Rappaport's broth (R25). The inoculum used with R25 was 0.005 ml, determined in an earlier study. Two incubation temperatures were employed with strontium chloride enrichment (37 and 43 degrees C). Rappaport's medium was incubated at 37 degrees C only. Elevated temperature enrichment at 43 degrees C improved the performance of strontium chloride B, but Rappaport's broth still gave significantly better results. This supports earlier studies on simplification of salmonella isolation and standardization of routine technique on a single enrichment medium: Rappaport broth (R25) incubated at 37 degrees C.

Bacteriological Techniques↗
PubMed1982

Kinetics of transfused neutrophils.

Rational neutrophil transfusion therapy depends upon an understanding of the in vivo fate of the transfused cells. The ability of transfused cells to circulate and to accumulate at sites of inflammation in the recipient is profoundly affected by abnormalities in the viability of the cells transfused as well as by abnormalities in the recipient. Autologous transfusion studies in normal subjects have shown that cells collected by centrifugation are kinetically nearly normal in vivo, whereas cells collected by filtration are markedly abnormal. Storage of either cell preparation for one day results in substantial additional abnormalities. Studies in animals have shown that recipient variables such as inflammation, endotoxin, or neutropenia significantly alter the kinetics of transfused normal cells and that the inflammatory site accumulation of cells in such animals is not reliably predicted by the blood kinetic measurements. Problems associated with labelling neutrophils for kinetic studies are discussed.

Adrenal Cortex Hormones↗