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Biomedical subjects

R C Cooksey

Publications and source records attributed to R C Cooksey.

At least 37 records · Page 2Linked to original sources

Identification of an epidemic strain of Acinetobacter baumannii using electrophoretic typing methods.

Nosocomial infections due to Acinetobacter baumannii dramatically increased in a Lebanese medical center following an outbreak of hostilities in Lebanon in 1984. The incidence of infection caused by this organism has remained high in this institution, thus requiring the implementation of a strain typing system to aid in infection control. Three methods were investigated for their utility in differentiating among a representative group of 36 nosocomial Acinetobacter baumannii isolates obtained over a 10 month period from specimens of hospitalized patients. Isolates were typed by antibiogram analyses, plasmid fingerprinting, and total cell protein profiles. Only three distinct total cell protein profiles were detected, with one pattern accounting for 26 (72.2%) of the isolates. However, eight different plasmid profiles were observed, with 20 (55.5%) isolates having the same profile. Eleven distinct antibiograms were seen with the most prevalent pattern occurring in 21 isolates. Twenty of the 21 (95%) isolates with the common antibiogram also had the same plasmid profile and total protein profile (44.4% of total isolates). The combination of these three typing methods was useful in tracing the spread of these organisms in the medical center. The data obtained suggest the distribution of a common strain among at least six wards of this hospital.

Acinetobacter↗

Antithyroid effects of coal-derived pollutants.

Endemic goiter in iodide-sufficient areas of the United States and Colombia has been linked to watersheds rich in coal and shale, which several reports suggest are the source of water-borne goitrogens. In this report the potential antithyroid activities of aqueous coal and shale extracts and of compounds identified in aqueous effluents from coal conversion processes were assayed in thyroid peroxidase (TPO) and thyroid slice systems. Aqueous extracts of coal and black shale were potent inhibitors of TPO or 125I organification by thyroid slices. The most abundant water-soluble compounds derived from coal are dihydroxy-phenols, thiocyanate, disulfides, and hydroxypyridines. The dihydroxyphenols resorcinol, 2-methylresorcinol, and 5-methylresorcinol (orcinol) were 26.7, 22.5, and 7.2 times more potent, respectively, than the antithyroid drug 6-propylthiouracil (PTU). Other dihydroxyphenols and thiocyanate were less potent but comparable in activity to PTU. All dihydroxypyridines and 3-hydroxypyridine produced inhibitory effects comparable to PTU. None of the disulfides inhibited TPO. The antiperoxidase effects of combinations of two dihydroxyphenols or one dihydroxyphenol and SCN were additive, whereas the effects of a combination of four dihydroxyphenols at threshold inhibitory concentrations were synergistic, resulting in net effects equivalent to or greater than the sum of the individual effects. Thus, antithyroid effects may be greatly amplified by exposure to multiple coal-derived goitrogens and could be many times that produced by any one of the contributing pollutants. These results demonstrate that potent water-borne goitrogens are derived from coal and shale and that their contamination of water supplies could pose a serious threat of thyroid disorders.

Animals↗

Antimicrobial susceptibility patterns of common and unusual species of enterococci causing infections in the United States. Enterococcal Study Group.

We collected 705 isolates of enterococci (1 per patient) from cultures of a variety of anatomic sites from patients at eight tertiary-care hospitals in six geographic regions of the United States. A total of 632 (90%) Enterococcus faecalis, 58 (8%) E. faecium, 5 E. gallinarum, 4 E. avium, 3 E. casseliflavus, 1 E. raffinosus, and 1 E. hirae isolate and 1 biochemical variant of E. faecalis were identified; 606 (86%) of these isolates were associated with clinical infections. The most common sites of isolation were the urinary tract (402 [57%]), nonsurgical wounds (94 [13%]), the bloodstream (74 [10%]), and surgical wounds (62 [9%]). High-level resistance to gentamicin or streptomycin or both was detected in 265 (38%) of the isolates. We identified two E. faecalis isolates resistant to vancomycin (MICs, 32 and 128 micrograms/ml) and 11 beta-lactamase-producing E. faecalis isolates. E. faecium isolates were significantly more resistant than E. faecalis isolates to penicillin, ampicillin, piperacillin, imipenem, and ciprofloxacin (P less than 0.001). The MICs for the 15 non-E. faecalis, non-E. faecium enterococci indicated variable resistance to ciprofloxacin and the penicillins. Antimicrobial susceptibility patterns vary among species of enterococci, and these organisms, while commonly resistant to high-level aminoglycosides, can also acquire resistance to vancomycin or the ability to produce beta-lactamase. Because of these diverse antimicrobial resistance mechanisms, successful treatment and control of enterococcal infections with current antimicrobial agents are becoming increasingly difficult.

Adolescent↗

Antithyroid and goitrogenic effects of millet: role of C-glycosylflavones.

Pearl millet [Pennisetum millet (L.) leeke] is the main source of food energy for the rural poor in many areas of the semiarid tropics. Epidemiological evidence suggests that millet may play a role in the genesis of endemic goiter in these areas, and sparse experimental data in rats support this suspicion. This study was undertaken to determine in vivo in rats and in vitro using porcine thyroid slices and a thyroid peroxidase (TPO) assay the goitrogenic and antithyroid effects of millet diets, extracts of millet, and certain pure compounds contained therein. For use in these studies, whole grain millet was progressively dehulled to yield successively four bran and four flour fractions in which direct analyses revealed progressively lower concentrations of C-glycosylflavones. In vivo feeding of bran fraction 1, that richest in C-glycosylflavones, led to a significant increase in thyroid weight and antithyroid effects. Feeding of bran fraction 2, the next richest in C-glycosylflavones, produced similar, but less marked, changes. In vitro studies of 125I metabolism using porcine thyroid slices indicated that extracts of bran fractions 1 and 2 were most potent, producing changes similar to those produced by methimazole (MMI). At a concentration of 60 mumol/L, glucosylvitexin, the major C-glycosylflavone present in millet, had effects comparable to those of 1 mumol/L MMI. Similarly, in studies of porcine TPO, extracts of bran fraction 1 caused pronounced (85%) inhibition of enzyme activity, and progressively less inhibition was induced by extracts of bran fractions 2, 3, and 4. Overall, the TPO-inhibiting activities of the various millet fractions closely correlated with their C-glycosylflavone concentrations. Three C-glycosylflavones present concentrations. Three C-glycosylflavones present in millet, glucosylvitexin, glycosylorientin, and vitexin, also inhibited TPO activity. Thus, in vivo and in vitro studies revealed that millet diets rich in C-glycosylflavones produce goitrogenic and antithyroid effects similar to those of certain other antithyroid agents and small doses of MMI. We conclude that in areas of iodine deficiency in which millet is a major component of the diet, its ingestion may contribute to the genesis of endemic goiter.

Animals↗

Thyroid function in neonates from goitrous and nongoitrous iodine-sufficient areas.

We compared thyroid function between newborns from goitrous and nongoitrous localities in which iodine intake has been supplemented since 1955. Cord serum samples were analyzed in 185 infants born during a 9-month period (1986-1987) in 2 goitrous and 1 nongoitrous localities of western Colombia. Urinary iodine was determined in all mothers before delivery. No significant differences were found among neonates of the 3 localities (Kruskal-Wallis test) for the various thyroid hormone values, and all values were within the normal range, although there was a trend in distribution of TSH to higher values in both goiter areas. Thyroid autoantibodies (antithyroglobulin and antithyroid microsomal) were negative in all neonates, and iodine intake, as indicated by urinary iodine, was adequate and similar among the mothers of the 3 groups. Those newborn infants with serum TSH values higher than 20 mU/L were reexamined 5-7 months later. At this time, all infants had lower serum TSH values and their serum free T4 index and T3 values were normal. Gestational age, weight, and height at birth were normal and also equal among the neonates in the 3 localities. These results indicate that neonates from goitrous iodine-sufficient areas have thyroid function similar to that of infants born in nongoitrous areas equally supplemented with iodine, and therefore, they are not more at risk to develop congenital hypothyroidism.

Colombia↗

Biotyping coagulase-negative staphylococci.

The biochemical profiles obtained with Staph-Ident (Analytab Products, Plainview, N.Y.) panels were combined with the results of adherence and synergistic hemolysis tests to define biotypes among 1,064 clinical isolates representing eight species of coagulase-negative staphylococci. The 672 isolates of Staphylococcus epidermidis were aligned in 69 of 144 potential biotypes in our scheme because of 18 different biochemical profiles and the eight physiologic subtypes. Isolates of most other species were in fewer biotypes because of more uniform adherence and synergistic hemolysis data, as well as fewer biochemical profiles. Since adherence and synergistic hemolysis may prove to be related to virulence and pathogenicity, biotyping with these test results would help evaluate the reliability of adherence and synergistic hemolysis as possible indices of the clinical significance of some of these organisms. When the antimicrobial susceptibility and plasmid profiles obtained on two clusters of S. epidermidis isolates were compared with the biotyping results, one cluster was not further differentiated by plasmid profiles, but was by antimicrobial profiles; the other cluster with only two biotypes was further divided into five distinct types by plasmid profiles but was not separated at all by antimicrobial profiles.

Bacterial Adhesion↗

Molecular hybridization versus isoelectric focusing to determine TEM-type beta-lactamases in gram-negative bacteria.

Isoelectric focusing and molecular hybridization with a TEM DNA probe were used to screen for TEM beta-lactamase in 328 bacterial isolates representing 11 gram-negative genera. The TEM enzyme was detected in 50% of isolates, and nine additional types of beta-lactamase could be identified in 36.9% of isolates. The TEM gene was detected in 53.6% of isolates. The results obtained by both methods were concordant in 92.7% of the entire sample. In situ colony hybridization with a specific probe therefore appears to be a convenient method to screen rapidly for the presence of homologous genetic sequences among a large number of isolates. Positive hybridization was observed for 16 isolates in which no TEM beta-lactamase was detected by isoelectric focusing. The significance of this hybridization remains to be determined.

DNA, Bacterial↗

Nucleic acid probes in clinical microbiology.

The infectious disease applications of nucleic acid probe have been described. In addition, the basic procedures of nucleic acid probe technology have been discussed, as have the factors affecting implementation of probe technology in diagnostic laboratories. Despite the questions raised, nucleic acid probes will become part of the diagnostic laboratory in the near future. Commercial interests are developing and marketing new probes, reagents, and kits which will expedite the employment of this technology. High-volume reference laboratories will first use probes as part of a battery of tests which will include ELISA and monoclonal antibody methods. In all probability, probes will replace methods: that have proven to be ineffective, difficult, or costly such as culturing for some enteric pathogens and Legionella, that require long incubation periods, such as mycobacteria, or that have high costs and low yields, such as virology.

Bacteria↗

Microplate phosphocellulose binding assay for aminoglycoside-modifying enzymes.

We modified the phosphocellulose binding assay for aminoglycoside-modifying enzymes (AMEs) by use of microdilution plates and a multichannel micropipette. Batteries of aminoglycoside substrates for screening organisms for the presence of AMEs as well as for subclassifying enzymes were prepared and stored in microdilution plates. When tested in parallel with the conventional tube reaction assay, the microplate assay yielded comparable radioactive counts and therefore equally correct identifications of AMEs in 32 isolates representing nine bacterial species. Other modifications, such as multichannel dispensing of crude enzyme preparations and radioisotopic precursors, provided a more rapid, convenient, and less expensive means of examining large collections of organisms for AMEs.

Acetyltransferases↗

Relatedness of tetracycline resistance plasmids among species of coagulase-negative staphylococci.

Four isolates of Staphylococcus aureus and 98 isolates of coagulase-negative staphylococci representing six species all obtained from endocervical cultures were examined for antimicrobial susceptibility and for the presence of plasmids. More than 80% of the isolates were susceptible to each of 12 antimicrobial agents tested, whereas only 33% were susceptible to penicillin G, 30% were susceptible to cadmium chloride, and 41% were susceptible to tetracycline. Although no species-related susceptibility or plasmid patterns were detected, 77 isolates contained at least one plasmid and 43 contained a plasmid similar in mass to a 2.7-megadalton tetracycline resistance plasmid previously reported in staphylococci. Association of tetracycline resistance with plasmids of this size in four species was determined from curing experiments. No plasmids homologous with the tetracycline resistance locus of the Escherichia coli plasmid pBR322 were found among 11 isolates examined by DNA hybridization. Homology with a 2.7-megadalton plasmids (pRC701) from an endocervical isolate of S. aureus, however, was apparent for 2.7-megadalton plasmids harbored by six isolates as well as with larger plasmids harbored by three isolates. Restriction analysis revealed that pRC701 shared structural identity with two plasmids of a similar mass from two species of coagulase-negative staphylococci as well as with a previously characterized tetracycline resistance plasmid originating in S. aureus.

Base Sequence↗

A gene probe for TEM type beta-lactamases.

A restriction fragment of plasmid pBR322 bearing the TEM-1 beta-lactamase structural gene was electroeluted from agarose gels after digestion with EcoRI and HinfI. The 1-kilobase fragment was 32P-labeled and used to examine genetic relationships with nucleic acids encoding seven other beta-lactamase classes. The probe hybridized only with TEM-2 and OXA-2 class plasmids.

DNA, Bacterial↗

In vitro measurement of antithyroid compounds and environmental goitrogens.

A specific, sensitive, and reproducible in vitro assay for antithyroid compounds and environmental goitrogens has been used to investigate antithyroid activity (AA) in small samples of water supplying 15 localities in endemic and nonendemic goiter areas of western Colombia. A significant positive correlation was observed between goiter prevalence and AA in water collected from the pipelines of these localities. Samples at the water source showed only borderline significance. No significant correlation was observed in waters between AA and total hardness (ppm) or concentrations of Ca, Mg, sulfates, chlorides, silicates, nitrates, and iodine. AA was also demonstrated by this in vitro assay in well water previously shown experimentally to be goitrogenic and that supplied the endemic goiter district of Candelaria town in western Colombia. In contrast, water from the well supplying the area of lower endemicity was found to possess little AA. These results provide experimental support for epidemiological observations that demonstrate a relationship between the sources of drinking water and goiter prevalence rates, and are consistent with previous findings indicating that organic antithyroid compounds contaminate water supplies in areas where goiter persists despite adequate iodine supplementation.

Animals↗

In vitro antimicrobial inhibition patterns of nutritionally variant streptococci.

Twenty-four isolates of nutritionally variant streptococci, previously categorized as species included among the viridans streptococci, were tested for susceptibility to 12 antimicrobial agents. Minimum inhibitory concentrations for these isolates and for a control group of viridans streptococci with no apparent nutritional deficiencies were determined in two microdilution systems. Pyridoxal hydrochloride, which enhances growth of the nutritional variants, was added to one of these microdilution systems but not to the other. An agar dilution method was also used to test the nutritionally variant isolates. Minimum inhibitory concentrations determined by the three methods compared favorably. Penicillin, nafcillin, methicillin, and clindamycin were less effective in vitro against the nutritional variants than were the other antibiotics tested. Streptomycin, however, was less effective against the control isolates.

Anti-Bacterial Agents↗

Physiological characterization of nutritionally variant streptococci.

Twenty-five isolates of nutritionally variant streptococci submitted to the Streptococcus Laboratory of the Center for Disease Control over a 2-year period were tested for growth requirements and for biochemical reactions. After they were recovered from storage in blood at -170 degrees C, all isolates grew within 48 h in both thioglycollate broth and Todd-Hewitt broth supplemented with 0.001% pyridoxal.HCl. They grew better in the latter, even though they all grew on unsupplemented infusion agar, anaerobe blood agar, and chopped meat-glucose medium. Biochemical patterns of the isolates resemble those of five viridans streptococcal species. Two isolates had patterns which did not resemble those of any viridans species. Biochemical reactions obtained with heart infusion broth base biochemicals and carbohydrate fermentation media compared favorably for an overall agreement rate of 86.5% for key tests. Lactic acid and acetic acid were the major fermentation products detected with gas-liquid chromatography.

Acetates↗

Evaluation of commercial latex agglutination reagents for grouping streptococci.

A total of 155 strains of beta-hemolytic streptococci were serologically grouped by conventional techniques (Lancefield extraction and capillary precipitin testing) and by latex agglutination (LA). Agreement between conventional and LA techniques was 97% when the instructions of the manufacturer for the LA technique were followed. Agreement of 99% was obtained when modified autoclave extracts were used as antigens in the LA procedure. A total of 82 strains of non-beta-hemolytic streptococci were also tested by conventional, prescribed LA, and modified autoclave procedures. The agreement between conventional techniques and both LA procedures was 76%. However, when serological cross-reactions in the conventional grouping procedures were considered as errors, the accuracy of identification of both LA procedures was 88% among the non-beta-hemolytic strains. Of 13 strains of Streptococcus bovis, 10 did not react with the LA group D reagent but were serogroup D by conventional techniques. More S. bovis strains were grouped by the LA technique when extracts of 20 ml of broth cultures were used as antigens; however, cross-reactions were observed with non-group D strains when this technique was applied to them.

Hemolysis↗

Presumptive identification of group A, B, and D streptococci on agar plate media.

Several presumptive tests were evaluated for their effectiveness in differentiating streptococci. When the tests were combined into a battery and the resulting reactions were interpreted as patterns, the overall presumptive identification rate was at least 97%. We used the hemolytic reaction, susceptibility to bacitracin and sulfamethoxazole plus trimethoprim (1.25 micrograms plus 23.75 micrograms), and standard CAMP reactions on sheep blood Trypticase soy agar, and bile-esculin and 6.5% NaCl agar tolerance tests with incubation in candle extinction jars. Subsequently, 98.9% of the group A; 95.3% of the group B; 100% of the beta-hemolytic non-group A, B, or D; 92.3% of group D enterococcal; 100% of the group D non-enterococcal; and 92.8% of the viridans streptococci were presumptively identified. We then used the hemolytic reactions, susceptibility of bacitracin and sulfamethoxazole-plus-trimethoprim disks, CAMP disk reactions on sheep blood Trypticase soy agar and bile-esculin and 6.5% NaCl agar tolerance tests with incubation in normal atmosphere. Subsequently, 98.1% of the group A; 98.6% of the group B; 99.2% of the beta hemolytic non-group A, B, or D; 97.5% of the group D entercoccal; 97.6% of the group D non-entercoccal; and 92.4% of the viridans strains were presumptively identified.

Aerobiosis↗

Antimicrobial susceptibility patterns of Streptococcus pneumoniae.

Fifty clinical isolates of Streptococcus pneumoniae received by the Streptococcus Laboratory of the Center for Disease Control from August 1976 through March 1977 and 50 pneumococcal strains retrieved from 13- to 16-year storage (originally isolated October 1961 through December 1964) were tested for susceptibility to 10 antimicrobial agents by disk-agar diffusion and agar dilution tests. No appreciable differences in susceptibility patterns were apparent between the two groups, and, except for one isolate, all were highly susceptible to every drug tested except gentamicin. This single isolate required higher drug concentrations to inhibit macroscopic growth and had corresponding decrements in zones of disk inhibition with penicillin, ampicillin, and cephalothin. An additional 43 pneumococci recently received from various areas of the United States and Canada were screened by a disk agar diffusion method for penicillin resistance. Four of these isolates had penicillin zone diameters <30 mm, and subsequent agar dilution test results showed that the penicillin minimum inhibitory concentrations were elevated with these organisms. Antimicrobial susceptibility patterns of pneumococci to antimicrobials other than penicillin and its analogs have not changed substantially in over a decade. However, due to the emergence of strains with decreased susceptibility to penicillin, the screening test for penicillin resistance in pneumococci, especially in isolates from spinal fluid and blood, could be clinically useful as an aid in selecting optimal therapy.

Cephalosporinase↗