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Biomedical subjects

W R Butler

Publications and source records attributed to W R Butler.

At least 55 records · Page 3Linked to original sources
PubMed1996

Automated DNA fingerprinting analysis of Mycobacterium tuberculosis using fluorescent detection of PCR products.

DNA fingerprints of Mycobacterium tuberculosis are produced by restriction fragment length polymorphism analysis of the insertion element IS6110. We modified a PCR-based subtyping method, mixed-linker PCR with fluorescent-labeled IS6110-specific oligonucleotides, to demonstrate rapid, automated, and unattended electrophoretic analysis. Variation in band sizing (normally occurring with fragment mobility), an artifact of lane-to-lane and gel-to-gel differences, was controlled with an internal lane standard, resulting in accurate and precise DNA sizing. By using this method, fingerprint analysis can be performed using actual fragment length rather than estimated position analysis.

Base Sequence↗
PubMed1996

The mtp40 gene is not present in all strains of Mycobacterium tuberculosis.

A multiple PCR-based assay that targets IS6110 and the mtp40 gene was evaluated for the rapid differentiation of Mycobacterium bovis and M. tuberculosis, two of the causative agents of tuberculosis. The IS6110 target is present in both species, whereas the mtp40 gene was thought to be specific for M.tuberculosis (P. Del Portillo, L.A. Murillo, and M.E. Patarroyo, J. Clin. Microbiol. 29:2163-2168, 1991). However, the mtp-40 gene is not present in all M. tuberculosis strains and, hence, is not useful for differentiating M.tuberculosis and M.bovis.

Bacterial Proteins↗
PubMed1996

Prolonged dominance of follicles and reduced viability of bovine oocytes.

Prolonging the lifespan of bovine follicles is known to result in reduced fertility after ovulation and insemination. In this study, the effect of prolonged development of follicles on oocyte viability was examined. In Expt 1, six cows in the aspirated-prolonged-follicle group received a vaginal progesterone releasing device on day 4 of the oestrous cycle (day 1 = ovulation) and prostaglandin F2 alpha (40 mg) on day 6. Ultrasound-guided follicular aspiration was performed on day 13. Six cows in the aspirated-growing-follicle group received prostaglandin F2 alpha on day 6, and follicular aspiration on day 7. In Expt 2, all cows were stimulated with 36 mg FSH-P to develop multiple large follicles for study. Three cows in the prolonged-multiple-follicle group received the same treatment as did cows in Expt 1, but were ovariectomized on day 13. Three cows in the growing-multiple-follicle group received prostaglandin F2 alpha on day 6 and were ovariectomized on day 7. Oocytes recovered in both experiments were stained to reveal the stage of nuclear development. All oocytes from aspirated-prolonged and prolonged-multiple follicles showed expanded cumulus cells and condensed chromatin dispersed in the ooplasm, with possible germinal vesicle breakdown. Oocytes from aspirated-growing and growing-multiple follicles showed compact cumulus cells and an intact germinal vesicle. Plasma concentrations of oestradiol in both growing follicle groups increased until oocyte recovery. Oestradiol in the aspirated-prolonged-follicle group increased after luteolysis on day 6 and remained high until follicular aspiration. In contrast, in the FSH-stimulated prolonged-multiple-follicle group, oestradiol fell to trace amounts on day 8 and remained low. Oestradiol concentrations in follicular fluid were consistent with plasma concentrations for all groups. Bovine oocytes from prolonged dominant follicles undergo premature maturation in vivo, which perhaps accounts for the poor fertility observed in other studies when oestrous synchronization with progestins prolongs follicle lifespan.

Animals↗
PubMed1996

Plasma and milk urea nitrogen in relation to pregnancy rate in lactating dairy cattle.

The objectives of this study were to relate concentrations of plasma (PUN) and milk (MUN) urea nitrogen to pregnancy rate in dairy cows and compare various methods of analysis and preparation of milk for measuring MUN. In two experiments, blood or milk samples were collected on the day of AI from Holstein cows (n = 160 and n = 155, respectively). Three methods of MUN analysis were compared. Two laboratory chemical procedures yielded similar results, whereas a quick dipstick method overestimated chemical analyses. Before and after milking strip samples had MUN concentrations equivalent to those in composite milk. Concentrations of PUN or MUN greater than 19 mg/dL were associated with decreased (P < .02) pregnancy rates (18 and 21 percentage point reduction in the two experiments). In two subset groups of cows (n = 51 and n = 23, respectively), plasma progesterone or MUN concentrations were monitored during the 5-d period after AI. Plasma progesterone concentrations increased similarly during the period for cows divided into low vs high PUN but were greater in pregnant than in nonpregnant cows on d 4 and 5 (P < .04). The MUN concentrations showed low within-cow variation (CV = 8%) but were lower in pregnant cows and had a decreasing trend over time compared with nonpregnant cows (P < .05). Based on this study, plasma and milk will yield similar results for monitoring urea nitrogen in dairy cows; PUN and MUN concentrations > 19 mg/dL were associated with approximately a 20 percentage point decrease in pregnancy rate after AI in lactating dairy cattle.

Analysis of Variance↗
PubMed1995

Food restriction, gonadotropins, and behavior in the lactating rat.

This study sought to quantify effects of undernutrition on behaviors and to relate these to gonadotropin and prolactin concentrations in the lactating dam. Dams were studied in a 2 x 3 factorial design with litter size and food intake as the two factors. Behavioral data were collected from each dam and her litter on day 9, day 14, and day 19 of lactation, and maternal blood samples collected. Plasma was analyzed for luteinizing hormone, follicle stimulating hormone and prolactin. On day 15 of lactation, percent time nursing, number of pups actively nursing, total number of pups nursing and dam location acted as mediating factors of the effect of diet group on plasma luteinizing hormone concentration. No such relationships were seen for plasma follicle stimulating hormone, and only nest condition score appeared to be a mediator for plasma prolactin concentration. In conclusion, this analysis suggests that food restriction indirectly influences plasma concentration of luteinizing hormone, but not follicle stimulating hormone, by changing maternal and pup behaviors. The relationship among diet, behavior and circulating prolactin was less clear.

Animals↗
PubMed1995

Multidrug-resistant Mycobacterium tuberculosis in an HIV dental clinic.

OBJECTIVE: To investigate possible transmission of multidrug-resistant tuberculosis (MDR-TB) in a dental setting. DESIGN: A retrospective, descriptive study of dental workers (DWs), patients, and practice characteristics. PATIENTS: Two dental workers (DW1 and DW2) with acquired immunodeficiency syndrome and MDR-TB. SETTING: A hospital-based (Hospital X) human immunodeficiency virus (HIV) dental clinic in New York City. METHODS: To identify dental patients with tuberculosis (TB), patients treated in the dental clinic at Hospital X during 1990 were cross-matched with those listed in the New York City Department of Health Tuberculosis Registry. Mycobacterium tuberculosis isolates from both DWs and from dental patients with TB were tested for antimicrobial susceptibility and typed by restriction fragment length polymorphism (RFLP) analysis. Infection control practices were reviewed. RESULTS: M tuberculosis isolates infecting DW1 and DW2 were resistant to isoniazid and rifampin and had identical RFLP patterns. DW1 and DW2 worked in close proximity to each other in a small HIV dental clinic in Hospital X during 1990. Of 472 patients treated in the dental clinic in 1990, 41 (8.7%) had culture-proven M tuberculosis infection. Of these 41, 5 had isolates with resistance patterns similar to both DWs; however, for four available isolates, the RFLP patterns were different from the patterns of the DWs. Sixteen of the 41 patients received dental treatment while potentially infectious. Dental patients were not routinely questioned about TB by dental staff, nor were all dental staff screened routinely for TB. No supplemental environmental measures for TB were employed in the dental clinic in 1990. CONCLUSIONS: Our investigation suggests that MDR-TB transmission may have occurred between two DWs in an HIV dental clinic. Opportunities for transmission of TB among dental staff and patients were identified. TB surveillance programs for DWs and appropriate infection control strategies, including worker education, are needed to monitor and minimize exposure to TB in dental settings providing care to patients at risk for TB.

Acquired Immunodeficiency Syndrome↗
PubMed1994

Rapid identification of mycolic acid patterns of mycobacteria by high-performance liquid chromatography using pattern recognition software and a Mycobacterium library.

Current methods for identifying mycobacteria by high-performance liquid chromatography (HPLC) require a visual assessment of the generated chromatographic data, which often involves time-consuming hand calculations and the use of flow charts. Our laboratory has developed a personal computer-based file containing patterns of mycolic acids detected in 45 species of Mycobacterium, including both slowly and rapidly growing species, as well as Tsukamurella paurometabolum and members of the genera Corynebacterium, Nocardia, Rhodococcus, and Gordona. The library was designed to be used in conjunction with a commercially available pattern recognition software package, Pirouette (Infometrix, Seattle, Wash.). Pirouette uses the K-nearest neighbor algorithm, a similarity-based classification method, to categorize unknown samples on the basis of their multivariate proximities to samples of a preassigned category. Multivariate proximity is calculated from peak height data, while peak heights are named by retention time matching. The system was tested for accuracy by using 24 species of Mycobacterium. Of the 1,333 strains evaluated, > or = 97% were correctly identified. Identification of M. tuberculosis (n = 649) was 99.85% accurate, and identification of the M. avium complex (n = 211) was > or = 98% accurate; > or = 95% of strains of both double-cluster and single-cluster M. gordonae (n = 47) were correctly identified. This system provides a rapid, highly reliable assessment of HPLC-generated chromatographic data for the identification of mycobacteria.

Algorithms↗
PubMed1994

Multiplex PCR assay specific for the multidrug-resistant strain W of Mycobacterium tuberculosis.

In 1991, a multidrug-resistant strain of Mycobacterium tuberculosis was isolated from eight people with tuberculosis at a state correctional facility in New York. This strain, which is designated strain W (IS6110 restriction fragment length polymorphism type 212072), was resistant to isoniazid, rifampin, ethambutol, streptomycin, kanamycin, ethionamide, and rifabutin. Since that outbreak, the W strain has been associated with outbreaks in five hospitals in the New York City area and is a continuing public health problem in the area. To be able to identify this strain rapidly, we developed a multiplex PCR assay which targets a direct repeat of IS6110 with a 556-bp intervening sequence (NTF-1). The amplification generates two amplicons from strain W, which indicate the presence and orientation of the NTF-1 sequence between the direct repeat of IS6110, and a third amplicon, which serves as an internal PCR control. The assay was evaluated with 193 isolates of M. tuberculosis, and all 48 strain W isolates among those 193 isolates were correctly identified.

Antitubercular Agents↗
PubMed1993

Mycobacterium celatum sp. nov.

A new slowly growing nonphotochromogenic Mycobacterium species of clinical importance is described. The biochemical characteristics of this organism were similar to those of Mycobacterium xenopi and members of the Mycobacterium avium complex. However, none of the strains reacted with commercially available genetic probes for the M. avium complex. The strains were resistant to most antituberculosis drugs. Multilocus enzyme electrophoresis revealed two original electrophoretic types, which was suggestive of new species. The strains contained alpha-, keto-, and dicarboxylic mycolates, as determined by thin-layer chromatography. A mycolic acid analysis by high-performance liquid chromatography revealed a chromatographic pattern similar to that of M. xenopi, but distinct from the patterns of previously described Mycobacterium species. Hexadecanoic and tuberculostearic acids were identified as the major cell wall fatty acids by gas-liquid chromatographic analysis; hexacosanoic acid was the major mycolic acid cleavage product, and 2-eicosanol was the major alcohol. Evaluation of the 16S rRNA sequence confirmed the phylogenetic position of the organism among the slowly growing Mycobacterium species. Cultures representing this new species have been deposited in the American Type Culture Collection as strains ATCC 51130 and ATCC 51131T (T = type strain). The name Mycobacterium celatum is proposed.

Bacterial Typing Techniques↗
PubMed1993

Rapid, amplification-based fingerprinting of Mycobacterium tuberculosis.

Insertion element IS6110 occurs in multiple copies throughout the Mycobacterium tuberculosis genome, and the variability of its insertion sites is the basis for the IS6110 restriction fragment length polymorphism (RFLP) method for typing. We describe a novel gene amplification method to assess the variability of the location of IS6110. A unilateral-nested polymerase chain reaction and hybridization procedure was used to measure the variability in the distances between IS6110 elements and copies of a major polymorphic tandem repeat sequence of M. tuberculosis. The pattern of amplicons produced could be used to cluster epidemiologically related strains of M. tuberculosis into groups which correlated with the groups formed using IS6110-RFLP typing. Reliable patterns can be generated directly from sputum specimens as well as from M. tuberculosis cultures. We designated the novel method as IS6110-ampliprinting.

Bacterial Typing Techniques↗
PubMed1993

Clinical significance, biochemical features, and susceptibility patterns of sporadic isolates of the Mycobacterium chelonae-like organism.

Mycobacterium chelonae-like organisms are nonpigmented rapidly growing mycobacteria whose clinical significance is unknown. We evaluated 87 sporadic isolates encountered in a clinical laboratory. Most isolates (62%) were respiratory; only 2 of 54 (4%) (both from patients with AIDS) were clinically significant. Among 33 nonrespiratory isolates, 20 of 33 (or 61%) were clinically significant. Clinical diseases included posttraumatic wound infections and catheter-related sepsis. Routine biochemical features included growth inhibition by 5% NaCl (100%), a smooth colony morphology (94%), positive 3-day arylsulfatase reaction (84%), no color or a light tan color on iron uptake (100%), and variable nitrate reduction (45%). Additional characteristics that helped to separate this group from M. chelonae and Mycobacterium abscessus were susceptibility to cephalothin (90%) and ciprofloxacin (100%), utilization of mannitol (94%) and citrate (83%) as carbon sources, and unique patterns of mycolic acid esters by high-performance liquid chromatography. This group was quite drug susceptible, with 100% of isolates inhibited by amikacin, imipenem, cefoxitin, cefmetazole, and the newer quinolones ciprofloxacin and ofloxacin. Three examples of this group, including a proposed type strain, have been deposited in the American Type Culture Collection.

Bacteriological Techniques↗
PubMed1993

Mixed-linker polymerase chain reaction: a new method for rapid fingerprinting of isolates of the Mycobacterium tuberculosis complex.

Rapid recognition of multidrug-resistant strains of Mycobacterium tuberculosis is a desirable goal for treatment of patients and protection of health care workers. DNA fingerprints produced with the insertion sequence IS6110 generate restriction fragment length polymorphism (RFLP) patterns that reliably identify M. tuberculosis complex strains. This report describes a rapid technique for RFLP typing using the polymerase chain reaction. The method uses one primer specific for IS6110 and a second primer complementary to a linker ligated to the restricted genomic DNA. In one strand the linker contains uracil in place of thymidine, and specific amplification is obtained by elimination of this strand with uracil N-glycosylase. Mixed-linker fingerprinting clearly differentiated multidrug-resistant isolates from 12 outbreaks and unambiguously assigned them to 26 RFLP groups.

Base Sequence↗
PubMed1993

Reduction of fertility and alteration of uterine pH in heifers fed excess ruminally degradable protein.

The objective of these studies was to elucidate the causes underlying the reduction of fertility associated with feeding of excess ruminally degradable protein to cattle. Holstein heifers were fed total mixed rations that either met (Normal) or exceeded (High) ruminally degradable protein requirements. In Exp. 1 heifers (n = 80) were inseminated at estrus after being fed the experimental diets for 4 wk. First-service conception rates were 82 and 61% for the Normal and High groups, respectively (P < .05). Plasma urea nitrogen (PUN) levels were elevated (10.2 +/- .17 vs 14.8 +/- .19 mg/dL) in the High group throughout the experiment (P < .05). In Exp. 2, heifers (n = 32) receiving the same diets as in Exp. 1 were fitted with catheters in a jugular vein, in the bladder, and in the uterus on either the day before expected estrus or d 6 after estrus. On estrus or d 7, plasma and urinary NH4 and urea nitrogen and uterine pH were determined at 4-h intervals for 24 h beginning just before feeding. Plasma NH4 was variable and did not differ between treatments or days of the estrous cycle. During the 24-h period, urinary urea nitrogen and PUN were elevated (P < .05) in the High groups and did not differ between days of the cycle. Urinary NH4 excretion was elevated (P < .05) during the 24-h period only in the High group on d 7. Uterine pH did not change over time after feeding.(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonia↗
PubMed1993

Alterations of pH in response to increased dietary protein in cattle are unique to the uterus.

This study was undertaken with two objectives: 1) to determine whether the effect of excess dietary protein on intrauterine pH in cattle is specific to the uterus or manifested in other bodily fluids and 2) to determine whether the effect of excess ruminally degradable protein on uterine pH can be ameliorated by substitution with a less-degradable protein source. Thirty-six Holstein cows in early lactation were fed isoenergetic total mixed rations that either 1) met undegradable intake protein (UIP) and degradable intake protein (DIP) requirements (Balanced), 2) met DIP requirements and exceeded UIP requirements by 25% (High UIP), or 3) met UIP requirements and exceeded DIP requirements by 25% (High DIP). After diets had been fed > or = 2 wk, uterine, blood, salivary, and urinary pH and plasma urea nitrogen were determined at estrus (d 0) and d 7. Plasma urea nitrogen (mg/dL) was not different between estrus and d 7 but was significantly affected by diet (Balanced, 16.1 +/- 2.3; High UIP, 19.2 +/- 1.6; High DIP, 22.3 +/- 2.6; P < .05). There was no effect of treatment on the pH of any fluid measured at estrus: intrauterine, blood, salivary, and urinary pH averaged 6.84 +/- .05, 7.39 +/- .01, 8.30 +/- .05, and 8.15 +/- .05, respectively. In contrast, on d 7, uterine pH was significantly lower in both high-protein groups, regardless of protein degradability (Balanced, 7.13 +/- .05; UIP, 6.95 +/- .04; DIP, 6.85 +/- .05; P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Social influences on reproductive development and fertility in female Djungarian hamsters (Phodopus campbelli).

Social influences on the sexual maturation of female Djungarian hamsters were investigated in two experiments. In the first experiment females were housed from weaning with an adult male, by themselves, or with a weanling sister. Maturation was accelerated in females housed with males as indicated by younger age at first ovulation, increased rates of ovarian and uterine growth, and lower LH levels at some ages. Maturation was delayed in females housed with sisters compared to those housed alone as measured by time of first ovulation and by lower estradiol levels at some ages. The most marked differences between groups occurred 8 to 12 days after weaning, suggesting that events during this period are particularly important in the social mediation of sexual maturation. In the second experiment the effects of reproductive suppression (caused by living with a sister) on the subsequent fertility of females housed with males were examined. If male-female pairs were housed in clean cages, no effects were observed; however, pairs housed in cages previously soiled by the female and her sister had fewer young surviving until 1 week of age despite no differences in the age of pregnancy onset or in the initial litter size. Thus, even cues present in unrenewed soiled bedding may have subtle but long lasting effects on reproductive function.

Animals↗