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Biomedical subjects

W J Martin

Publications and source records attributed to W J Martin.

At least 19 recordsLinked to original sources

Recovery of anaerobic bacteria from pediatric patients. A one-year experience.

During 1975, 115 speciments from the soft tissues, body fluids, and purulent sites in 105 pediatric ward and nursery patients were submitted to the clinical laboratory for anaerobic study, and 75% of the cultures were positive. In most instance specimens were collected in anaerobic transport tubes containing peptone yeast extract and glucose. In the laboratory, samples were processed by the anaerobic holding jar method. Nineteen different anaerobic organisms were isolated; the average number of species per specimen was 1.8 Bacteroides sp were the most common isolates (45 strains). Other isolates included 44 anaerobic cocci (15 of which were Peptostreptococcus sp), 22 Propionibacterium sp, ten Fusobacterium sp, nine Clostridium sp, eight Veillonella sp, six Bifidobacterium sp, and four Eubacterium sp. The main sources of anaerobic isolates included 52 gastric aspirates, 46 soft tissue sites, 14 blood specimens, 12 CSF samples, and 12 samples from the external ear canals. Forty-one (43%) of the 86 specimens that contained anaerobic bacteria also had aerobic organisms present.

Adolescent

API and Minitek systems in identification of clinical isolates of anaerobic gram-negative bacilli and Clostridium species.

A comparison of the API and Minitek methods of biochemical testing was made on a variety of anaerobic bacteria. Although API and Minitek results were not compared to more standardized or conventional procedures of identification, multiple repeat testing of the two systems was done on routine clinical isolates and known organisms to determine (i) whether the reactions were reliably consistent, (ii) the ease of reading the two systems with respect to the frequency of questionable results, and (iii) the percentage of routine clinical isolates for which each system yielded an identification. The Minitek system gave a much lower incidence of difficult to interpret reactions. The two systems were comparable in terms of reproducibility and capability of yielding an identification of the anaerobic gram-negative bacilli and Clostridium species, but were unsatisfactory for routine use on most of the other anaerobic bacteria isolated.

Bacterial Infections

Comparison of rates of virus isolation from leukocyte populations separated from blood by conventional and Ficoll-Paque/Macrodex methods.

One hundred fifty-two blood specimens, largely from immunocompromised patients, were collected in heparinized Vacutainer tubes and divided into paired aliquots of equal volume. Buffy-coat preparations, containing mixed leukocyte and separate mononuclear and polymorphonuclear leukocyte populations were obtained by treatment of blood with conventional and Ficoll-Paque/Macrodex (F-P/M) methods. The development of cytopathic effect in monolayers of WI-38 fibroblasts inoculated with cell suspensions derived from the two methods was used to assess virus infectivity. Twice as many virus isolations were obtained using F-P/M. Of those viruses isolated by both conventional and F-P/M, the development of cytopathic effect was more extensive using the latter method. Moreover, a greater variety of viruses was isolated using F-P/M method, as compared to the conventional method. The F-P/M method is no more time consuming than conventional procedures, is readily adaptable for use in the diagnostic virology laboratory, requires only minimal additional cost, and is a particularly suitable and effective means of monitoring viremia.

Adenoviruses, Human

Abnormal interaction of alpha 1-antitrypsin and leukocyte elastolytic activity in patients with chronic obstructive pulmonary disease.

The interaction of serum alpha 1-antitrypsin with leukocyte elastolytic activity was examined in 71 patients with chronic obstructive pulmonary disease and 46 normal subjects. Residual elastolytic activity was present despite adequate amounts of alpha1-antitrypsin in 33 of the 71 patients compared to 6 of the 46 normal control subjects (P less than 0.001). This elastolytic activity was completely abolished by a specific human leukocyte elastase inhibitor. A crossover study designed to detect the source of this abnormal activity revealed that the plasma fraction containing alpha 1-antitrypsin was responsible for inadequate inhibition in 30 of the 39 cases. The residual elastolytic activity from a given patient did not correlate with the serum alpha 1-antitrypsin concentration, alpha 1-antitrypsin phenotype, or history of smoking. Our data suggest that the abnormal interaction of alhpa 1-antitrypsin with leukocyte elastolytic activity is an important additional variable in the gensis of chronic obstructive pulmonary disease.

Female

Relative opsonic and protective activities of antibodies against K1, O and lipid A antigens of Escherichia coli.

The K1 Escherichia coli capsular antigen has been implicated as a virulence factor because of the frequency of isolation of K1 containing strains from certain invasive human infections. In the study of the interaction between K1 strains, normal human polymorphonuclear cells (PMNs) and fresh human serum, we have found varying susceptibility to phagocytosis and killing; thus, the in vitro opsonophagocytic and in vivo protective role of K1, somatic O and core glycolipid antibodies remain unclear. We have therefore examined strains of E. coli with defined susceptibility to phagocytosis by normal PMNs and sera and compared the effect of K1, somatic O and lipid A antibodies in opsonophagocytic tests and mouse protection experiments. K1 E. coli strains demonstrating relative resistance to phagocytosis and killing were effectively opsonized only with specific K1 capsular antisera. Similarly, K1 capsular antisera, but not anti-O or lipid A antisera, also provided protection in mice challenged with a LD100 of K1 E. coli that were "resistant" to phagocytosis. The ability of purified capsular antigens from Neisseria meningitidis group B and K1 "resistant" E. coli to inhibit the phagocytosis of a "sensitive" non-K1 and a K1 E. coli strain of "intermediate" susceptibility to opsonophagocytosis was also investigated. Purified K1 and group B capsular antigens were able to block specific capsular-antibody mediated opsonophagocytosis, yet these capsular antigens failed to inhibit the phagocytosis of non-K1 "sensitive" or K1 "intermediate" E. coli. These studies suggest that K1 antibodies are obligatory for the in vitro and in vivo opsonophagocytosis of "resistant" K1 E. coli and that the K1 antigen must remain in situ on the bacterial surface to exert an anti-phagocytic effect.

Animals

Lung tumor-reactive cytotoxic lymphocytes generated in mixed lymphocyte reaction between C3HfeB/HeN (H-2kb) and C3H/HeN (H-2k) strain mice.

The tumor-associated transplantation antigen expressed by several transplacentally induced lung tumors of C3HfeB/HeN mice (H-2kb haplotype) has previously been shown to exist as a normal tissue alloantigen in mice of known H-2k and H-2a haplotypes. This antigen is not expressed in normal tissues of C3HfeB/HeN mice but is expressed in C3H/HeN mice, the strain from which the C3HfeB/HeN mice were originally derived. The present study indicates that spleen cells from C3HfeB/HeN and C3H/HeN mice respond reciprocally in the mixed lymphocyte reaction. Cytotoxic T lymphocytes specific for the tumor-associated alloantigen can be readily generated in mixed lymphocyte reactions in which spleen cells from C3HfeB/HeN mice are reacted with x-irradiated spleen cells from C3H/HeN or A strain mice. These cells are effective in suppressing the growth of the C3HfeB/HeN-derived lung tumor 85 in x-irradiated syngeneic recipients.

Animals

Anaerobic and aerobic bacteriology of acute conjunctivitis.

Aerobic and anaerobic cultures and clinical data were obtained from 131 patients presenting with acute conjunctivitis. Similar cultures were obtained from 60 noninflamed individuals. Anaerobes were isolated from 66 patients, 51 times in mixed culture with aerobes and 15 times (11.5%) as the only isolates. Aerobes only were recovered in 54 patients. The organisms found to be statistically significantly more commonly recovered from eyes with conjunctivitis were Staphylococcus aureus, Hemophilus influenzae, Peptostreptococcus, and Propionibacterium acnes. No statistical difference was present between the bacterial flora of the eyes with unilateral conjunctivitis when compared to their uninvolved mates in 20 cases in which only one eye was involved.

Adolescent

Aerobic and anaerobic bacterial flora of the maternal cervix and newborn gastric fluid and conjunctiva: a prospective study.

Aerobic and anaerobic cultures were performed on cervical swabs from 35 mothers and their newborn infants' gastric aspirates and conjunctival sacs. Four hundred seventy-nine isolates were obtained; 287 were aerobes and 192 were anaerobes. The cervical and gastric cultures overall yielded a similar flora, but this was not always the case when one compared mothers with their own newborn infants. The conjunctival cultures yielded about half of the number of bacteria per specimen that the gastric contents yielded, and a repeat conjunctival culture done 48 hours later showed a marked reduction in the number of potentially pathogenic bacteria isolated. The predominant aerobes isolated were Staphylococcus epidermidis, diphtheroids, viridans streptococci and Haemophilus vaginalis. The predominant anaerobes were the Bacteroides fragilis group, Propionibacterium acnes, Peptococcus, other Bacteroides and Peptostreptococcus organisms. Statistical analysis revealed significant positive correlation between the isolation of certain groups of organisms and increased duration of pregnancy, increased baby's weight, and prolonged duration of labor.

Bacteria

Lung tumor-associated derepressed alloantigen coded for by the K region of the H-2 major histocompatibility complex.

Transplacental induction of lung tumors in C3HfeB/HeN (C3Hf) strain mice can be readily achieved with the carcinogen 1-ethyl-1-nitrosourea. Several of these tumors express, as a tumor-associated transplantation antigen (TATA), a normal tissue alloantigen present in strain A and C3H/HeN (C3H) mice. In the present study it was shown that the tumor-associated alloantigen on the C3Hf-derived lung tumor 85 was present in all mice of H-2(a) and H-2(k) haplotypes tested and in CBA (532) strain mice (H-2(ka) haplotype). Studies using congenic-resistant and recombinant strains of mice indicated that the genetic locus controlling the expression of this antigen was either within or to the left of the H-2K region of the major histocompatibility complex (MHC). Thus the antigen was expressed in B10.A (4R) mice (kkbbbb MHC haplotype) but not in B10 (bbbbbb) or B10.AQR mice (qkkddd). The antigen was expressed in all tissues tested of C3H and A strain mice. It was not detected on any tissue tested including embryo tissue of C3Hf mice or mice of MHC haplotype other than H-2(k) or H-2(a). Because C3Hf strain mice were originally derived from C3H strain mice (H-2(k)), the MHC haplotype of C3Hf mice has been provisionally designated H-2(kb). The finding of a tumor-associated change in the expression of a H-2K region-coded antigen is consistent with the concept that MHC-coded antigens may act as targets for immunological surveillance of tumors.

Animals

Histocompatibility difference between C3HfeB/HeN and C3H/HeN mice: tumour induced in C3HfeB/HeN mice expresses C3H/HeN-associated alloantigen.

A transplacentally induced lung tumour of C3HfeB/HeN mouse origin expresses, as a tumour-associated antigen, a normal tissue component of strain A mice. The genetic locus coding for this alloantigen has been shown to be linked to the H-2 major histocompatibility complex. In the present study we demonstrate that this antigen is also expressed on normal tissues of C3H/HeN mice. Skin grafts exchanged between C3HfeB/HeN and C3H/HeN mice are reciprocally rejected at approximately 3 weeks after grafting. C3HfeB/HeN mice were derived from C3H/HeN mice in 1945. These strains have apparently deviated since then in their genetic regulation of the expression of the MHC-linked genetic locus. The finding of the C3H/HeN-associated antigen on a C3HfeB/HeN mouse-derived lung tumour indicates that this deviation is reversible.

Animals

Modified agar dilution method for rapid antibiotic susceptibility testing of anaerobic bacteria.

A simplified method has been developed for agar dilution antimicrobial susceptibility testing of anaerobic bacteria, designed to economize on time and money when only a few isolates need to be tested. The procedure is based on the principle of using filter paper disks as carriers of the antibiotic and 35- by 10-mm petri dishes which, when inoculated with the Steers replicator, can test up to four organisms per plate. The procedure was run in parallel with conventional agar dilution techniques and showed 95% agreement to within one dilution for all minimal inhibitory concentrations recorded on fresh anaerobic isolates from clinical specimens. The technique was further simplified by using commercially available antibiotic-containing disks, thereby alleviating the tedious and time-consuming procedure of preparing the disks. The data indicated that 48- to 72-h diffusion periods were sufficient to achieve a uniform concentration of the antibiotic in the petri plate and that the antibiotics were stable at room temperature for that period of time. In terms of applicability and relevance to the needs of the clinical microbiology laboratory, the modified agar dilution method for rapid antimicrobial susceptibility testing of individual anaerobic isolates was found to be superior to the broth dilution method since it was easier to read and required considerably less set up time.

Agar

Comparison of the Minitek test system with a conventional screening procedure for identification of Enterobacteriaceae.

A Minitek basic 9 disc/11 test system plus arabinose, inositol and rhamnose was compared with a conventional six tube screening system for identification of members of the family Enterobacteriaceae. Correlation varied from 88.3 to 100% with the various substrates when studied against 737 strains of these and other gram-negative bacteria recently isolated in our clinical laboratory. Two discs, citrate and ornithine, which originally gave a correlation of 88.3 and 95.5% respectively were modified so that a final correlation of 99.4 and 96.3% was obtained. The average correlation using the eleven discs was 97%.

Bacteriological Techniques

Hemagglutinin-specific cytotoxic T-cell response during influenza infection.

Specific cytotoxic thymus-derived (T) lymphocytes were detected in the cervical lymph nodes and spleen during influenza infection of mice. The cytotoxic T cells can distinguish target cells infected with different influenza A subtypes. Infection with parent viruses and their recombinant progeny possessing the hemagglutinin of one parent and the neuraminidase of the other demonstrated that significant cytotoxicity occurred only when the hemagglutinin of the immunizing viruses was the same as that of the virus used to infect the target cell. In addition to this specific cytotoxic response to the major surface antigen, a cross-reactive response could be detected when the relatively nonpermissive L cell was used as the target cell. These results indicate there is a specific cytotoxic T-cell response to the surface hemagglutinin, and a cross-reactive cytotoxic response, not directed to the hemagglutinin, during influenza infection. The cytotoxic T-cell response specific for the hemagglutinin antigen may play an important role in in vivo immunity to influenza.

Animals

Specificity studies on cytotoxic thymus-derived lymphocytes reactive with influenza virus-infected cells: evidence for dual recognition of H-2 and viral hemagglutinin antigens.

Cytotoxic thymus-derived (T) lymphocytes were readily detected in spleens of mice inoculated intranasally with mouse-adapted A/Port Chalmers (H3N2), A/England (H3N2), A/PR/8 (H0n1), and B/Hong Kong influenza viruses. T-cell-mediated lysis of H-2 compatible target cells infected with the strain of virus used to immunize the mice was considerably higher than lysis of either syngeneic cells infected with a different strain of influenza virus or allogeneic cells infected with the immunizing strain of influenza virus. The findings that cytotoxic lymphocytes can distinguish minor antigenic variants among influenza viruses and that lysis depends on H-2 histocompatibility between lymphocyte and target cell support the concept of dual recognition of visual and H-2 histocompatibility antigens in T-cell-mediated antiviral immunity.

Animals