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M C Roberts

Publications and source records attributed to M C Roberts.

At least 37 records · Page 2Linked to original sources

Identification of the conjugative mef gene in clinical Acinetobacter junii and Neisseria gonorrhoeae isolates.

The mef gene, originally described for gram-positive organisms and coding for an efflux pump, has been identified in clinical isolates of Acinetobacter junii and Neisseria gonorrhoeae. These strains could transfer the mef gene at frequencies ranging from 10(-6) to 10(-9) into one or more of the following recipients: gram-negative Moraxella catarrhalis, Neisseria perflava/sicca and Neisseria mucosa and gram-positive Enterococcus faecalis. Three Streptococcus pneumoniae strains could transfer the mef gene into Eikenella corrodens, Haemophilus influenzae, Kingella denitrificans, M. catarrhalis, Neisseria meningitidis, N. perflava/sicca, and N. mucosa at similar frequencies. The mef gene can thus be transferred to and expressed in a variety of gram-negative recipients.

Acinetobacter↗

A novel multiresistant Streptococcus pneumoniae serogroup 19 clone from Washington State identified by pulsed-field gel electrophoresis and restriction fragment length patterns.

In 1997, a cluster of multiresistant invasive serogroup 19 pneumococcus infections, including two fatalities, was reported in Washington State. Further investigation identified other cases. Fourteen Washington Streptococcus pneumoniae isolates, four from Alaska, and eight isolates from eastern Canada with reduced penicillin susceptibility (MIC of > or =1 microg/ml) were included in the study. Pulsed-field gel electrophoresis (PFGE) with ApaI, SacII, and SmaI restriction enzymes and IS1167 and mef restriction fragment length polymorphism (RFLP) pattern analysis were performed. Twenty of the 26 isolates had identical or related PFGE patterns, with two or all three enzymes, and identical or related IS1167 RFLP patterns, indicating that they were genetically related. These 20 isolates contained the mef gene conferring erythromycin resistance and had identical mef RFLP patterns. The PFGE and RFLP patterns were distinct from those of six multiresistant clones previously described and suggest that a new multiresistant clone has appeared in Washington, Alaska, and eastern Canada. This newly characterized clone should be included in the Pneumococcal Molecular Epidemiology Network.

Anti-Bacterial Agents↗

Genistein augments prostaglandin-induced recovery of barrier function in ischemia-injured porcine ileum.

We have previously shown that PGE(2) enhances recovery of transmucosal resistance (R) in ischemia-injured porcine ileum via a mechanism involving chloride secretion. Because the tyrosine kinase inhibitor genistein amplifies cAMP-induced Cl(-) secretion, we postulated that genistein would augment PGE(2)-induced recovery of R. Porcine ileum subjected to 45 min of ischemia was mounted in Ussing chambers, and R and mucosal-to-serosal fluxes of [(3)H]N-formyl-methionyl-leucyl phenylalanine (FMLP) and [(3)H]mannitol were monitored as indicators of recovery of barrier function. Treatment with genistein (10(-4) M) and PGE(2) (10(-6) M) resulted in synergistic elevations in R and additive reductions in mucosal-to-serosal fluxes of [(3)H]FMLP and [(3)H]mannitol, whereas treatment with genistein alone had no effect. Treatment of injured tissues with genistein and either 8-bromo-cAMP (10(-4) M) or cGMP (10(-4) M) resulted in synergistic increases in R. However, treatment of tissues with genistein and the protein kinase C (PKC) agonist phorbol myristate acetate (10(-5)-10(-6) M) had no effect on R. Genistein augments recovery of R in the presence of cAMP or cGMP but not in the presence of PKC agonists.

8-Bromo Cyclic Adenosine Monophosphate↗

The surface coat of infective larvae of Trichinella spiralis.

The surface coat of the infective larvae of the parasitic nematode Trichinella spiralis was characterized with respect to its biophysical properties, morphology and composition. Labelling of larvae with the fluorescent surface probe PKH26 was lost after activation (by incubation in mammalian medium containing trypsin and bile), or following pronase treatment. Electron microscopical examination revealed that pronase treatment resulted in the loss of an amorphous surface layer only, further demonstrating the specificity of PKH26 for the larval surface coat. Surface coat shedding was inhibited by sodium azide and carbonyl cyanide, or by incubation of larvae at 4 degrees C, suggesting the shedding process required metabolic energy. Pre-labelled, unactivated larvae demonstrated continuous slow surface coat shedding and could be re-labelled with PKH26, indicating that the shed coat is replaced in these parasites. However, pre-labelled larvae which were activated failed to re-label with the probe, suggesting that activation provides an irreversible trigger for surface changes. PKH26, therefore, is a useful marker for larval activation. Examination of the shed coat material by scanning electron microscopy revealed 2 types of morphologies; one comprising thin multilaminate sheets and the other of amorphous material with ridges producing a fingerprint-like motif. Western- and lectin-blotting of the shed coat material demonstrated 2 prominent entities; a 90 kDa glycoprotein, which bound Datura stramonium agglutinin and was resistant to N- and O-glycanase treatment and a 47-60 kDa set of protein(s). Analysis of the surface lipids by electrospray mass spectometry revealed the presence of lysophosphatidic acid (lysoPA, C14:2) and an unidentifiable component of 339.4 Da. These two lipids constituted 36.9% and 36% by mass of surface coat lipids respectively. The presence of lysoPA was confirmed by thin layer chromatography, which also detected phosphatidic acid (PA). The polar lipids detected in solvent rinses of intact parasites by electrospray mass spectrometry were PI (C48:4), PE (C40:4 and C38:4), PS (C40:4), lysoPC (C20:2 and C18:2) and lysoPA (C14:2). These observations are discussed with respect to the role of the surface coat and its shedding in the T. spiralis host-parasite relationship.

Animals↗

Characterization of a multidrug-resistant clone of invasive Streptococcus pneumoniae serotype 6B in Alaska by using pulsed-field gel electrophoresis and PspA serotyping.

Antimicrobial susceptibility, pneumococcal surface protein A (PspA) serotyping, and pulsed-field gel electrophoresis (PFGE) were used to evaluate clonal relatedness among 66 invasive isolates of Streptococcus pneumoniae serotype 6B collected during 1982-1996 from patients in Alaska. Thirty-seven (56%) of the isolates had penicillin minimal inhibitory concentration values >/=0.125 microgram/mL and were resistant to at least 1 other antibiotic. Fourteen PspA serotypes were observed; PspA 16 was the most common (35%). Forty-five (68%) of the 66 isolates shared common and highly related PFGE patterns using 3 enzymes. Twenty-six (58%) of the isolates with common PFGE patterns were from Native Alaskan children </=2 years of age residing in 1 region of Alaska. Alaskan serotype 6B had distinct PFGE patterns, compared with the South African 6B-8 and Spanish 6B-2 multidrug-resistant clones, suggesting that the Alaskan 6B isolates were distinct from these other pneumococcal 6B clones but were genetically related to each other [corrected].

Adolescent↗

Host range of the ermF rRNA methylase gene in bacteria of human and animal origin.

We screened 183 different clinical anaerobic and aerobic bacteria isolated from humans and other animals for the presence of the ermF gene using a polymerase chain reaction (PCR) assay. The ermF gene was detected in 107 (58%) clinical isolates, including 42 (61%) of 69 gram-positive bacteria and 65 (57%) of 114 gram-negative bacteria. Twenty-five ATCC isolates were also tested; 20 (80%) carried the ermF gene. The gene products from the ermF PCR from four isolates were sequenced and showed 95-99% nucleotide homology with the ermF gene and 98-99% amino acid homology with the gene product. Eleven (58%) of the 19 gram-negative donors tested were able to transfer the ermF gene. All nine (100%) of the gram-positive donors tested transferred the ermF gene, using either Enterococcus faecalis or Haemophilus influenzae as the recipients.

Animals↗

A variety of gram-positive bacteria carry mobile mef genes.

The mefE gene codes for a membrane bound efflux protein, which confers resistance to macrolides, and has been identified in Streptococcus pneumoniae. A variety of gram-positive organisms were examined. Twenty-six isolates of S. pneumoniae carried mefE and were resistant to erythromycin (MIC of 2-16 mg/L). Two additional isolates of Emr S. pneumoniae carried both ermB and mefE(MIC of 16-128 mg/L). One Micrococcus luteus, one Corynebacterium jeikeium, three Corynebacterium spp., two viridans streptococci and seven Enterocccus spp. also carried mef genes. It was possible to move the mef gene from all 11 S. pneumoniae tested to susceptible S. pneumoniae and/or Enterococcus faecalis recipients. The addition of DNase (1 g/L) did not affect the gene transfer. It was also possible to move the mef gene from donor Enterococcus spp., viridans streptococci, M. luteus, C. jeikeium and Corynebacterium spp. to E. faecalis recipients. Transconjugant isolates were resistant to erythromycin (MIC = 16 mg/L). Hybridization with a labelled mef oligonucleotide probe against Southern blots and bacterial dot blots confirmed the presence of the mef genes. This is the first time that a mobile mef gene has been identified in four different genera, from three distinct geographical locations.

Anti-Bacterial Agents↗

Mobile elements carrying ermF and tetQ genes in gram-positive and gram-negative bacteria.

Bacteroides spp. conjugative transposon Tn5030 is 150 kb which includes a 43 kb characterized region containing a number of defined genes and an open reading frame (ORF). The 43 kb region is organized with the ORF1 immediately upstream from the ermF gene, coding for an rRNA methylase, then an unknown 20 kb region downstream followed by the tetQ gene (coding for a ribosomal protection protein) then the rteA and rteB genes. The role of ORF1 is unclear; rteA is a putative sensor and rteB a regulator. Thirty-seven (62%) of 60 isolates, representing one gram-positive anaerobic and 13 gram-negative anaerobic species, co-transferred the ermFand tetQ genes to an unrelated Enterococcus faecalis recipient. We used the polymerase chain reaction to show the linkage between ORF1, ermF, tetQ, rteA and rteB. Our data suggest that the ORF1 gene product may participate in the transfer of the ermF gene with or without the ORF1-rteB region and has homology to bacterial transposases. Isolates that co-transferred the ermF and tetQ genes carried and transferred the rteB gene, suggesting that the rteB gene product may be important in transfer of the 43 kb ORF1-rteB region to E. faecalis. The rteB gene product is not required when ermF is transferred independently of tetQ.

Bacterial Proteins↗

The characteristics of intestinal injury peripheral to strangulating obstruction lesions in the equine small intestine.

Recent studies suggest that horses requiring surgical correction of strangulating intestinal obstruction may develop post operative complications as a result of ischaemia/reperfusion injury. Therefore, the mucosal and serosal margins of resected small intestine from 9 horses with small intestinal strangulating lesions were examined for evidence of ischaemia/reperfusion injury. Severe mucosal injury and marked elevations in myeloperoxidase activity were detected at ileal resection margins (n = 4), whereas the mucosa from proximal jejunal (n = 9) and distal jejunal (n = 5) resection margins was normal. However, the serosa from jejunal resection margins had evidence of haemorrhage and oedema, and the proximal jejunal serosa had significantly increased numbers of neutrophils. Histological injury in ileal stumps is indicative of the inability fully to resect the ileum in horses with distal small intestinal strangulations. One of 4 horses subjected to ileal resection was subjected to euthanasia and found to have a necrotic ileal stump. Evidence of serosal injury and neutrophil infiltration in the proximal jejunal resection margins may predispose horses to post operative adhesions. Four of 8 horses discharged from the hospital suffered from recurrent colic in the post operative period.

Anastomosis, Surgical↗

Role of duodenal reflux in nonglandular gastric ulcer disease of the mature horse.

Gastric contents were sampled in horses via nasogastric tube to determine changes in pH and bile salt concentrations during feeding and fasting periods. The horses were rotated through 4 feeding protocols. (1) hay; (2) hay with twice daily grain meals; (3) and (4) fasting preceded by either hay only or hay and grain. Sequential, hourly samples were collected from 3 horses prepared with gastric cannulas. Horses were fed hay twice daily and grain mix either twice daily or in small aliquots dispensed every 90 min. The horses were sampled during normal feeding or after 14 h of feed deprivation. Gastric pH values varied with time, but there was no significant difference between the feeding protocols or the fasting period on mean pH. Bile salt concentrations in fasted animals averaged 0.23-0.44 mmol/l with individual samples greater than 0.9 mmol/l. The bile salt concentrations in fed animals were consistently below 0.2 mmol/l. The effect of bile salt and acid on the stratified squamous gastric mucosa was tested in vitro. Mucosa, stripped of muscle and serosal layers, was mounted in Ussing chambers and the electrical potential difference (PD) across the tissue recorded. Sodium taurocholate or deoxycholate (0.3 mmol/l, bile salt) and/or HCl were added to the mucosal bathing solutions. The bile salt alone had no significant effect. Addition of acid (pH 2.5) to control tissues caused a decrease in the PD, which recovered within 20 min. Addition of acid to tissues exposed to bile salts resulted in a significant decrease in the PD, which did not recover. We conclude that combinations of bile salts and acid are more injurious to the stratified squamous gastric mucosa of the equine than acid alone. Concentrations of bile salts and acid sufficient to alter the electrolyte transport function of this mucosa can be found in the gastric contents of horses deprived of feed for as little as 14 h.

Animals↗

Erythromycin-resistant Neisseria gonorrhoeae and oral commensal Neisseria spp. carry known rRNA methylase genes.

Two Neisseria gonorrhoeae isolates from Seattle and two isolates from Uruguay were resistant to erythromycin (MIC, 4 to 16 microg/ml) and had reduced susceptibility to azithromycin (MIC, 1 to 4 microg/ml) due to the presence of the self-mobile rRNA methylase gene(s) ermF or ermB and ermF. The two Seattle isolates and one isolate from Uruguay were multiresistant, carrying either the 25.2-MDa tetM-containing plasmid (Seattle) or a beta-lactamase plasmid (Uruguay). Sixteen commensal Neisseria isolates (10 Neisseria perflava-N. sicca, 2 N. flava, and 4 N. mucosa) for which erythromycin MICs were 4 to 16 microg/ml were shown to carry one or more known rRNA methylase genes, including ermB, ermC, and/or ermF. Many of these isolates also were multiresistant and carried the tetM gene. This is the first time that a complete transposon or a complete conjugative transposon carrying an antibiotic resistance gene has been described for the genus Neisseria.

Amino Acid Sequence↗

Nomenclature for new tetracycline resistance determinants.

Letters of the English alphabet have heretofore been used to name tetracycline resistance determinants. Since all 26 letters have now been used, a nomenclature employing numerals is recommended for future determinants, and one laboratory has offered to coordinate the assignment of numerals.

Terminology as Topic↗

Prostaglandin-induced recovery of barrier function in porcine ileum is triggered by chloride secretion.

We have previously shown that PGI2 and PGE2 have a synergistic role in restoring electrical transepithelial resistance (R) in ischemia-injured porcine ileum via the second messengers Ca2+ and cAMP. Because Ca2+ and cAMP stimulate Cl- secretion, we assessed the role of PG-induced Cl- secretion in recovery of R. Mucosa from porcine ileum subjected to ischemia for 45 min was mounted in Ussing chambers and bathed in indomethacin and Ringer solution. Addition of PGs stimulated a twofold increase in R, which was preceded by elevations in short-circuit current (increase of 25 microA/cm2). The PG-induced effect on R was partially inhibited with bumetanide, an inhibitor of Cl- secretion. The remaining elevations in R were similar in magnitude to those induced in ischemic tissues by amiloride, an inhibitor of Na+ absorption. Treatment with 10(-4) M 8-bromo-cGMP or 300 mosM mucosal urea resulted in elevations in R similar to those attained with PG treatment. PGs signal recovery of R via induction of Cl- secretion and inhibition of Na+ absorption, possibly by establishing a transmucosal osmotic gradient.

16,16-Dimethylprostaglandin E2↗

Anxiety disorders among abstinent alcohol-dependent patients.

The Spielberger State-Trait Anxiety Index (STAI) and screening questions for anxiety disorders from the Structured Clinical Interview for DSM-IV Disorders were used to assess 146 alcohol-dependent patients on admission for inpatient treatment and just before discharge, after about three to four weeks of abstinence. Eighty-four patients (58 percent) reported excessive anxiety on admission, and 32 patients (22 percent) met DSM-IV criteria for specific anxiety disorders. After three to four weeks of abstinence, both state and trait STAI scores declined significantly, and only 15 patients (10 percent) met criteria for specific anxiety disorders. Anxiety levels on admission were significantly related to severity of alcohol dependence and recent life events but not to severity of withdrawal symptoms.

Adult↗

Training in clinical child psychology: doing it right.

Discusses (a) what roles the specialty of clinical child psychology fulfills and how societal and professional changes have enhanced the need for the specialty, (b) how the field defines itself, (c) how models of training are conceptualized for the specialty, and (d) how some training programs implement specialty training with broad, interdisciplinary components. Clinical child psychology is a professional field of research and practice that, when adequate training is provided, properly deserves a places as a specialty. The dangers of overspecialization and narrowness are more likely present in traditional clinical (adult) psychology than in clinical child psychology, especially when the clinical child training is done in a broadly comprehensive and integrated manner.

Adolescent↗