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Biomedical subjects

M G Johnson

Publications and source records attributed to M G Johnson.

At least 73 records · Page 4Linked to original sources

Surgical audit: variations in wound infection rates of individual surgeons.

A prospective survey of 1086 general surgical in-patients was carried out in West Dorset district to determine the incidence of postoperative infections for individual surgeons. A total of 79 cases were diagnosed as having become infected, resulting in an overall infection rate of 7.3%. For all cases the infection rate for individual surgeons varied from 3.9 to 14.6% (P less than 0.005; chi 2 test). For the clean operative category only, the infection rate varied from 2.9 to 20.0% (P less than 0.0005; chi 2 test). The results demonstrate the value of routine surgical audit in elaborating the variation in clinical outcomes for individual surgeons. Clinical audit can identify the need for extra supervision and training of junior staff and peer review for senior clinicians.

General Surgery↗

Purification and analysis of the major components of chum salmon protamine contained in insulin formulations using high-performance liquid chromatography.

A simple high-performance liquid chromatographic method has been developed for the rapid purification and analysis of protamine components contained in insulin formulations. Only a single step is needed to separate peptides whose compositions, sizes, and unusual isoelectric points (pI 13.8) are nearly identical. The method involves their isocratic separation on a reversed-phase column using a pH 2 phosphate buffer and a low acetonitrile content as an eluant. The purified chum salmon components were analyzed by amino acid analysis, solid-phase amino acid sequencing, carboxypeptidase B digests, insulin complexation analysis, and a mass spectrophotometric procedure which gives an accurate mass of the intact peptides. This HPLC purification technique may also be applicable to protamines and other highly basic peptides isolated from other sources.

Amino Acid Sequence↗

Monoclonal antibody specific for Listeria monocytogenes, Listeria innocua, and Listeria welshimeri.

Eight hundred fifty-nine murine hybridomas were produced from eight fusions, and 27 were characterized for secretion of antibodies reactive to Listeria monocytogenes. One monoclonal antibody (MAb), P5C9, reacted with all test strains of L. monocytogenes (31 of 31), L. innocua (3 of 3), and L. welshimeri (1 of 1) but not with any strains of the other four Listeria species or with any of 22 gram-positive or 11 gram-negative species of bacteria when tested in microtiter and dot blot enzyme immunoassays. Of the other 26 antibodies, 20 reacted with either L. monocytogenes Scott A or V7 and with some or all of the other six Listeria species but also cross-reacted with some or all of the non-Listeria bacteria tested. MAb P5C9 is of the immunoglobulin G1 murine subclass. In Western blot (immunoblot) analyses, this MAb reacted with a single antigen with a molecular weight of 18,500, and it is shared in common with all three reactive species, L. monocytogenes, L. innocua, and L. welshimeri. This antigen was extracted with detergent and appeared to be cell bound.

Animals↗

Petite colony formation by Listeria monocytogenes and Listeria species grown on esculin-containing agar.

Several strains of Listeria species formed petite-sized colonies from parent stock cultures when grown on agar media containing 0.2-1% (w/v) esculin. This was observed in Listeria monocytogenes (7/22 strains), L. innocua (1/3), L. grayi (1/1), L. seeligeri (1/3), and L. welshimeri (1/1), but not in L. ivanovii (0/1) and L. murrayi (0/1). This phenomenon was only observed on agar media that contained esculin. All petite isolates had biotyping profiles identical to their larger, normal-sized counterpart isolates. Normal and petite-sized isolates from two L. monocytogenes strains, Scott A and V7, were pathogenic to immunosuppressed white mice. On media containing 0.5% (w/v) esculin + ferric iron, Listeria cultures produced colony diameters intermediate in size between those of normal and petite cultures. When pregrown in glucose broth, all petite isolates demonstrated visible beta-glucosidase (esculinase) activity within 5 min, while the normal-sized isolates showed beta-glucosidase activity only after at least 20-70 min. This evidence suggests that cells forming petite colonies are beta-glucosidase constitutive variants within the parent population, while cells that form normal-sized colonies are inducible for beta-glucosidase (esculinase) activity. A possible role for the esculin hydrolysis product, esculetin, in causing petite colony formation is discussed.

Animals↗

Inhibition of Listeria monocytogenes growth by the lactoperoxidase-thiocyanate-H2O2 antimicrobial system.

The lactoperoxidase-thiocyanate-H2O2 system (LP system), consisting of lactoperoxidase (0.37 U/ml), KSCN (0.3 mM), and H2O2 (0.3 mM), delayed but did not prevent growth of L. monocytogenes Scott A at 5, 10, 20, and 30 degrees C in broth and at 20 degrees C in milk. The net lag periods determined spectrophotometrically varied inversely with temperature and were shorter at 5 and 10 degrees C for cultures from shaken versus from statically grown inocula. Lag periods for cultures from shaken and statically grown inocula, respectively, were 73 and 98 h at 5 degrees C, 22 and 32 h at 10 degrees C, both 8.9 h at 20 degrees C, and both 2.8 h at 30 degrees C. After the lag periods, the maximum specific growth rates were similar for each of the three treatments (complete LP system, H2O2 alone, or control broth) at 5, 10, and 20 degrees C and were 0.06 to 0.08, 0.09 to 0.1, and 0.32 to 0.36/h, respectively. At 20 degrees C in sterile reconstituted skim milk, the LP system restricted growth of Scott A, with log CFU counts per ml at 0, 36, and 68 h being 5.7, 6.4 and 7.9 (versus 5.7, 9.8, and 11.2 for controls). Possible explanations for the decreased lag times observed for cultures from aerobically grown inocula are discussed.

Animals↗

Persistence of Listeria monocytogenes in yogurt as determined by direct plating and enrichment methods.

Listeria Selective Isolation Agar (LSI) and Modified Acriflavin Ceftazidime Esculin Agar (MACE) were compared to McBride Listeria Agar minus Blood (MLA-B) for ability to recover Listeria monocytogenes Scott A cells inoculated into commercial yogurt, pH 4.1, Yogurt was stored at 5 degrees C and sampled periodically over a 12 day period. LSI, MACE and MLA-B inhibited the growth of the two yogurt organisms but LSI and MACE gave better inhibition of other separately tested Gram-positive bacteria likely to be present in other fermented foods. Acid-stressed Listeria monocytogenes Scott A cells were optimally recovered by enrichment at 5 degrees C for 5-18 days in 0.1 M phosphate buffer, pH 7.2, followed by transfer to tryptic soy broth +0.5% yeast extract at 30 degrees C for 2 days. At low inoculum levels (10(2) cells/g yogurt), they were not detectable by direct plating or enrichment of yogurt after day 0. At high inoculum levels (10(7) cells/g yogurt), they were detectable up to day 6 but not at day 9 by direct plating on MLA-B, LSI or MACE with log counts per gram of yogurt being about 10 fold higher on LSI than on MACE or MLA-B. The above enrichment procedure permitted recovery on MLA-B, LSI, or MACE of viable Listeria cells from the day 9 samples found negative by direct plating.

Animals↗

Drugs, alcohol and road accidents in Tasmania.

Drug analyses were performed on 200 blood samples that were taken for alcohol analysis from road users in Tasmania. Alcohol at a concentration of above 0.5 g was found in 75% of the samples, and other drugs were found in 17% of the samples. Cannabis was the most prevalent of these other drugs: it was detected in 6% of road users; benzodiazepine drugs were detected in 5% of road users; and barbiturate drugs were detected in 2% of road users. Alcohol was found in 50% and other drugs were found in 25%, of drivers, riders and pedestrians who were involved in road accidents that were serious enough to cause death or injury. In addition to alcohol, other drugs may be making a significant contribution to road accidents because all the drugs that were identified are capable of impairing psychomotor performance. Of particular concern is the prevalence of cannabis, which is an illegal drug, and barbiturate drugs, which are now prescribed rarely. A well-controlled study is required to quantitate the contribution of drugs other than alcohol to road accidents. In the meantime, drivers should be warned that drugs that depress the central nervous system can be expected to impair driving ability and to increase the risk of an accident.

Accidents, Traffic↗

Analysis of a mass colorectal cancer screening program for cost-effectiveness.

Mass screening of the general population with the stool blood test is a costly means of increasing the incidence of colorectal cancer identification; however, the test is worthwhile as a screening tool in patients at risk. We found the incidence of positive results to increase with age, as does the risk of colorectal cancer. Further efforts to increase the incidence of colorectal cancer detection should be directed toward increasing the awareness of primary care physicians and improving their screening practices. We stress that high-risk populations should be given special attention and the use of flexible proctosigmoidoscopes should be encouraged.

Colonic Neoplasms↗

Decreased glucose stimulation threshold, enhanced insulin secretion, and increased beta cell coupling in islets of prolactin-treated rats.

In order to determine the effect of lactogen on insulin secretion and junctional coupling among islet beta cells, ovine prolactin (oPRL) was infused by Alzet minipumps into female rats for 4 days. This treatment produced an oPRL level of 994 +/- 122 ng/ml which, combined with residual rat PRL (rPRL) (12 +/- 2 ng/ml), represented nearly a 20-fold increase from control (rPRL: 53 +/- 17 ng/ml). In addition, plasma insulin was increased nearly 50% (control: 21.9 +/- 3 microU/ml; experimental: 30.3 +/- 3 microU/ml; p less than 0.05). When pancreata from lactogen-treated and control animals were perfused with linear 30-200 mg/dl glucose gradients, the apparent glucose threshold for insulin secretion in the experimental group was nearly 33% lower than that of the controls (i.e., 70 +/- 4.6 mg/dl vs. 104 +/- 7.5 mg/dl; p less than 0.01). The oPRL treatment also increased dye coupling among beta cells. Central cells in islets isolated from lactogen-treated and control animals were injected with Lucifer Yellow CH to estimate the extent of gap junctional coupling. There was nearly a twofold increase in the projected area of dye transfer per injection in the experimental vs. the controls: 4,607 +/- 575 micron 2 vs. 2,302 +/- 474 micron 2, respectively; p less than 0.02. The effects of oPRL decreased the apparent glucose threshold for insulin release, increased the above-threshold glucose-induced insulin secretion, and increased the extent of dye coupling among beta cells. These changes in insulin secretion and dye coupling closely resemble those observed in islets from pregnant rats.

Animals↗

Localization of quantitative changes in pulmonary beta-receptors in ovalbumin-sensitized guinea pigs.

Impaired beta-receptor function has been postulated as one factor contributing to airway hyperreactivity in asthmatic patients. Although numerous indirect studies have cast doubt on this theory, none of these previous investigations has been able to directly measure changes in beta-receptor number on intrapulmonary structures capable of affecting the physiologic changes seen in this disease state. To help clarify the intrapulmonary location of such changes, a model of allergic bronchoconstriction was prepared by sensitizing guinea pigs to ovalbumin intraperitoneally (ip) 2 wk prior to testing (Group S). A second group of animals was sensitized to ovalbumin, then 2 wk later partially desensitized (Group D) during a 4- to 6-wk period by repeated exposure to increasing doses of nebulized ovalbumin with epinephrine rescue. Control animals received ip administered and nebulized normal saline alone. Pulmonary function assessed by plethysmography revealed an increase in airway resistance to 294 +/- 42% (SE) of control in Group S (p less than 0.005) and a decrease in dynamic compliance to 76 +/- 8% of control in Group D and 39 +/- 10% of control in Group S (p less than 0.002) after exposure to nebulized ovalbumin. Using L-[3H] dihydroalprenolol ([3H] DHA), beta-receptors were autoradiographically localized and quantitated in lung sections from all 3 groups. Significant decreases (p less than 0.02) in 3H-DHA binding were noted in alveolar and conducting airway epithelium, and bronchiolar and vascular smooth muscle in ovalbumin-exposed animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Sensitivity to bile salts of Shigella flexneri sublethally heat stressed in buffer or broth.

Batch cultures of Shigella flexneri M4243 were grown at 37 degrees C in broth to early stationary phase, washed, and heated at 50 degrees C in 0.1 M phosphate buffer (pH 7.0). Cells were surface plated on a tryptic phytone glucose agar (TPGA), TPGA with 0.15 or 0.85% bile salts no. 3 (TPGA-BS 0.15 or TPGA-BS 0.85), or TPGA with 0.25 or 0.50% sodium deoxycholate (TPGA-DC 0.25 or TPGA-DC 0.50). Cells sampled after no heating produced colony counts on TPGA-BS 0.85 or on TPGA-DC 0.50 that were no more than about 0.5 log lower than for unheated cell samples plated on TPGA. Cells heated at 50 degrees C for 30 min produced colony counts on TPGA-DC 0.50 or on TPGA-BS 0.85 that were about 1.5 logs lower than on TPGA. Cells heated for 30 min and shifted to TPG broth at 37 degrees C to allow resuscitation required about 2 h to regain tolerance to 0.85% BS. However, heated cells resuscitated on solid TPGA at 35 degrees C before being challenged with overlays of TPGA-BS 0.85 or TPGA-DC 0.50 required 6 to 8 h on TPGA to regain tolerance to 0.85% BS or 0.50% DC. To regain tolerance to overlays of 0.15% BS or 0.25% DC, heated cells required resuscitation periods on TPGA of about 2 or 2 to 6 h, respectively. Cells heated in TPG broth and sampled after no heating produced colony counts on TPGA that were about 1.5 logs lower than for unheated cell suspensions, suggesting greater apparent injury when heat stressed in broth than in buffer.(ABSTRACT TRUNCATED AT 250 WORDS)

Bile Acids and Salts↗

Isolation and characterization of a genetic variant of bovine proinsulin.

A genetic variant of bovine proinsulin has been isolated using preparative reverse-phase HPLC. The new proinsulin (bovine proinsulin II) differs from the known proinsulin (bovine proinsulin I) by a single amino acid residue at position C-48 in the connecting peptide. The amino acid replacement is a leucine substitution for proline. The two proinsulins were found in a ratio of approximately 9:1, proinsulin I: proinsulin II. No chemical or biological differences were observed for the two proinsulins other than their different elution times on reverse-phase HPLC.

Amino Acid Sequence↗

Distribution and quantitative developmental changes in guinea pig pulmonary beta-receptors.

Studies using tissue homogenates have demonstrated an increase in pulmonary beta-receptors during development. However, techniques using disrupted tissue have not permitted the precise anatomic localization of pulmonary beta-receptors or identification of structures where increases occur. Using L-[3H]dihydroalprenolol, beta-receptors were radioautographically localized and quantitated in sections of newborn (NB) and adult (A) guinea pig lung. Scatchard analysis showed a single class of binding sites with a maximum binding capacity of 189 +/- 3 (NB) and 305 +/- 37 (A) fmol X mg-1 protein (P less than 0.02). Binding was of high affinity with the dissociation constant (Kd) = 1.46 +/- 0.2 (NB) and 1.26 +/- 0.3 (A) nM (NS). The majority of beta-receptors were localized in alveolar wall and airway epithelia (alveolar much greater than bronchiolar greater than bronchial) (P less than 0.0001). Airway and vascular smooth muscle had significantly fewer demonstrable beta-receptors. The increased number of beta-receptors in the adult appeared to be due primarily to a 2.0 +/- 0.12-fold increase in alveolar wall and airway epithelia as opposed to only a 1.3 +/- 0.18-fold increase in the already low number in airway and vascular smooth muscle (P less than 0.05). While apparent receptor density may not necessarily correlate with physiological response or importance, radioautographic localization of pulmonary beta-receptors may significantly enhance our understanding of their role in normal and pathologic states.

Aging↗

Pancreatic polypeptide from rat pancreas.

Pancreatic polypeptide (PP) has been isolated from rat pancreas by gel filtration, ion exchange chromatography, and high performance liquid chromatography. The isolation was monitored with a RIA, using antibody to the carboxyl-terminal hexapeptide of bovine PP. Rat PP contains 36 amino acids and is similar in composition to PP from other mammalian sources. The single methionine residue in the peptide appears to oxidize easily to the sulfoxide, thereby giving rise to two immunoactive peaks on high performance liquid chromatography. Reduction to the native peptide can be accomplished with mercaptoethanol. The PP content of rat pancreas is about 2 mg/kg. The amino acid sequence of rat PP is Ala-Pro-Leu-Glu-Pro-Met-Tyr-Pro-Gly-Asp- Tyr-Ala-Thr-His-Glu-Gln-Arg-Ala-Gln-Tyr-Glu-Thr-Gln-Leu-Arg-Arg-Tyr-Ile- Asn-Thr-Leu-Thr-Arg-Pro-Arg-Tyr-NH2. This sequence preserves characteristics necessary for stabilization of the compact globular conformation found in avian PP.

Amino Acid Sequence↗

Zygomycosis caused by Cunninghamella bertholletiae: mycologic aspects.

Two isolates of Cunninghamella bertholletiae from confirmed cases of human zygomycosis were studied, and their sexual, asexual, and physiologic characteristics described. Minimal inhibitory concentrations for these two isolates, as well as for six other clinical isolates of C bertholletiae, were determined for amphotericin B, flucytosine, and miconazole using a standardized agar dilution technique. All isolates were found to be susceptible to miconazole, but resistant to flucytosine and amphotericin B.

Amphotericin B↗

Effects of pH shifts, bile salts, and glucose on sporulation of Clostridium perfringens NCTC 8798.

The sporulation of Clostridium perfringens NCTC 8798 was studied after exposing vegetative cells to: pH values of 1.5 to 8.0 in fluid thioglycolate broth (for 2h) and then transferring them to Duncan-Strong (DS) sporulation medium; sodium cholate or sodium deoxycholate (0.3 to 6.5 mM) in DS medium; or Rhia-Solberg medium with 0.4% (wt/wt) starch, glucose, or both added at 0 to 55 mM. At pH 1.5, no culturable heat-resistant spores were formed. For cells exposed to pH 3.0, 4.0, 5.0, or 6.0, increases in heat-resistant spores were not seen until after a lag of 12 to 13 h, whereas the lag was only 2 to 3 h for cells exposed to pH 7.0 or 8.0. Maximal spore crops were produced after only 6 to 8 h for cells exposed to pH 7 or 8, but 16 to 18 h was required for production of maximal spore crops by cells exposed to the lower-pH media. The addition of sodium cholate (3.5 to 6.5 mM) to DS medium only slightly reduced the culturable heat-resistant spore count from 1.9 X 10(7) to 3 X 10(6)/ml. The addition of 1.8 mM or more sodium deoxycholate reduced the culturable heat-resistant spore count to less than 10/ ml. When either starch or glucose alone was added to Rhia-Solberg medium there was no production of culturable heat-resistant spores, but a combination of 0.4% (wt/wt) starch and 4.4 mM glucose yielded 6 X 10(5) spores/ml. The spore production remained at this level for glucose concentrations of 6 to 22 mM, but then declined to about 3 X 10(3) spores per ml at higher concentrations.

Cholic Acids↗