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Biomedical subjects

M C Hudson

Publications and source records attributed to M C Hudson.

26 records · Page 2Linked to original sources
PubMed1991

Use of the polymerase chain reaction in detection of culturable and nonculturable Vibrio vulnificus cells.

Vibrio vulnificus is a human pathogen associated with consumption of raw oysters. During the colder months the organism apparently enters a viable but nonculturable state and thus cannot be cultured by ordinary bacteriological methods. For this reason, another means of detecting this bacterium is necessary. In the present study we utilized the polymerase chain reaction (PCR) to detect V. vulnificus DNA, thus eliminating the problem of nonculturability. DNA from both culturable and nonculturable cells of V. vulnificus was amplified by PCR with primers flanking a 340-bp fragment of the cytotoxin-hemolysin gene. As little as 72 pg of DNA from culturable cells and 31 ng of DNA from nonculturable cells could be detected. Fifty cycles of a two-step reaction (30 s [each] at 94 and 65 degrees C) were found to be optimal as well as more time efficient than the three-step PCR. The total procedure from the point of DNA extraction to observation on a gel required less than 8 h. Possible reasons for the difficulties encountered in amplifying DNA from nonculturable cells, e.g., gene rearrangement or loss of the hemolysin gene, are discussed.

Base Sequence↗
PubMed1990

Regulation of expression of Streptococcus mutans genes important to virulence.

Studies were initiated to investigate the regulation of Streptococcus mutans genes which are believed to be important to virulence. Operon fusions were constructed between S. mutans gene regulatory regions and a promoterless chloramphenicol acetyltransferase gene (cat) found on the plasmid pMH109. Specifically, fusions were generated between cat and the S. mutans genes encoding fructosyltransferase (ftf) and the glucosyltransferase B/C (gtfB/C) operon. Constructs were confirmed by restriction enzyme analysis, and the fusions were subcloned into the integration vehicle pVA891. Following generation of multimeric DNA, recombinant plasmids were introduced into the s. mutans genome by Campbell-type insertion, resulting in single-copy operon fusions. Chloramphenicol acetyltransferase specific activities were used to monitor the expression of the S. mutans gtfB/C operon and ftf determinants. The expression of these genes is increased by the presence of sucrose and is followed by a rapid decline in expression over time. Additionally, expression of the gtfB/C operon is increased in S. mutans cells bound to artificial tooth pellicles.

Chloramphenicol O-Acetyltransferase↗
PubMed1986

Differential utilization of Staphylococcus aureus promoter sequences by Escherichia coli and Bacillus subtilis.

Promoter-cloning plasmids were constructed and have been used to isolate transcriptionally active DNA fragments from Staphylococcus aureus. The plasmids contain a chloramphenicol acetyltransferase (CAT) gene of Gram-positive (G+) origin which lacks both its promoter and the sequence responsible for CAT inducibility. The ability of S. aureus promoters to direct CAT expression in Escherichia coli and Bacillus subtilis was examined. Two classes of staphylococcal promoter sequences have been obtained. Class I DNA fragments direct CAT expression in S. aureus, B. subtilis, and E. coli, while class II DNA sequences direct CAT expression only in the G+ hosts.

Acetyltransferases↗
PubMedDate not supplied

Evaluation of standard surgical preparation performed on superficial dermal abrasions.

OBJECTIVES: To determine the difference, if any, between the reduction of bacteria on contaminated normal skin and contaminated superficially abraded skin following standard surgical preparations at clinically relevant time points after injury. DESIGN: Prospective animal study. SETTING: Laboratory. SUBJECTS: Thirty-two New Zealand white rabbits. INTERVENTION: Two sites, two by two centimeters, one abraded and one nonabraded (control), were studied on each rabbit. Both were inoculated with encapsulated Staphylococcus aureus strain Wood 46. Four six-millimeter punch biopsies were obtained after inoculation, immediately before surgical scrub, and five minutes and then two hours after completion of the surgical scrub. The rabbits were divided into four cohort groups with surgical scrubs performed at six, twelve, twenty-four, and forty-eight hours after inoculation. Bacterial counts were determined. MAIN OUTCOME MEASUREMENTS: Numbers of bacteria on surgical sites. RESULTS: Before surgical preparation, the amount of bacteria on the normal skin (control sites) dropped significantly (p<0.02) except in the six-hour group (p<0.20). At the abraded skin sites, the bacteria flourished. The surgical scrub dropped bacterial counts at both the abraded and nonabraded skin sites significantly (p<0.05) except for the abraded site in the twenty-four-hour group (p<0.08). However in the twelve-, twenty-four-, and forty-eight-hour groups, the bacterial counts (colony-forming units) were still markedly elevated (>1x10(5) at abraded sites) when compared with the nonabraded skin sites (p<0.008) at the respective time intervals. Only at the six-hour interval were the bacterial counts reduced similarly at both the abraded and nonabraded skin sites. CONCLUSIONS: In a rabbit model the standard surgical preparation using povidone-iodine at six hours after inoculation is effective in reducing the bacterial count on abraded skin to that of surgically prepared nonabraded skin. Beyond that time, the standard surgical preparation is ineffective in reducing counts to those of nonabraded skin at similar time intervals.

Animals↗