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Biomedical subjects

R J Wallace

Publications and source records attributed to R J Wallace.

At least 55 records · Page 3Linked to original sources

Pre- and postgame macronutrient intake of a group of elite Australian football players.

This study describes pre- and postcompetition mean energy and macro nutrient intakes of 40 elite Australian Football players. Carbohydrate intake, expressed both as a percentage of total energy intake (En%) and as grams per kilogram of body mass (g/kg BM). Pregame carbohydrate intake (53.6% En) was significantly greater (p < .01) than postgame (49.7% En). However, expressed as g/kg BM, pre- and postgame macronutrient intakes did not differ significantly. Protein and fat intakes (as g/kg BM) fell within guidelines, whereas energy intake (13.2 MJ/day or 153.8 kJ/kg BM) was lower than expected. Results suggest that for athletes engaging in endurance team sports where body mass and energy requirements vary considerably, carbohydrate recommendations are more appropriately expressed as g/kg BM rather than En %.

Adult↗

Influence of 1-[(E)-2-(2-methyl-4-nitrophenyl)diaz-1-enyl]pyrrolidine-2-carboxylic acid and diphenyliodonium chloride on ruminal protein metabolism and ruminal microorganisms.

The effects of 1-[(E)-2-(2-methyl-4-nitrophenyl)diaz-1-enyl]pyrrolidine-2-carboxy lic acid (LY29) and diphenyliodonium chloride (DIC) on the degradation of protein to ammonia were determined in a mixed rumen microbial population taken from sheep on a grass hay-concentrate diet. Both compounds decreased NH3 production by inhibiting deamination of amino acids. LY29, but not DIC, inhibited growth of the high-activity ammonia-producing species, Clostridium aminophilum and Clostridium sticklandii.

Animal Feed↗

Evidence from molecular fingerprinting of limited spread of drug-resistant tuberculosis in Texas.

To determine the contribution of recent transmission to spread of drug-resistant tuberculosis in Texas, we performed IS6110-based and pTBN12-based restriction fragment length polymorphism (RFLP) analyses on Mycobacterium tuberculosis isolates. Isolates collected from 201 patients in Texas between 1992 and 1994 were studied. The distribution of cases was strikingly focal. All cases were reported from 35 of the 254 counties in Texas, and 74% (148 of 201) were reported from only 9 counties. One hundred sixty-one (80%) of the patients had M. tuberculosis isolates with unique RFLP patterns, and 41 (20%) patients were in 20 clusters, each comprising 2 to 3 patients. The largest number of cases of drug-resistant tuberculosis were reported in counties bordering Mexico, but the percentage of clustered cases was highest in northeast Texas and in counties that included the cities of Dallas, Fort Worth, and Houston. Compared to nonclustered patients, clustered patients were more likely to be African American and to have been born in the United States. Clustered patients were significantly more likely to be from the same geographic area, and clustered patients from the same geographic area were more likely to have isolates with identical drug susceptibility patterns, suggesting that they were linked by recent transmission. In 11 of 20 clusters, clustered patients were from geographically separate regions, and most isolates did not have identical drug susceptibility patterns, suggesting that tuberculosis was contracted from a common source in the remote past. Based on the low percentage of clustered cases and the small cluster size, we conclude that there is no evidence for the extensive transmission of drug-resistant tuberculosis in Texas.

Adolescent↗

Multisite reproducibility of results obtained by the broth microdilution method for susceptibility testing of Mycobacterium abscessus, Mycobacterium chelonae, and Mycobacterium fortuitum.

A multicenter study was conducted to assess the interlaboratory reproducibility of broth microdilution testing of the more common rapidly growing pathogenic mycobacteria. Ten isolates (four Mycobacterium fortuitum group, three Mycobacterium abscessus, and three Mycobacterium chelonae isolates) were tested against amikacin, cefoxitin, ciprofloxacin, clarithromycin, doxycycline, imipenem, sulfamethoxazole, and tobramycin (M. chelonae only) in four laboratories. At each site, isolates were tested three times on each of three separate days (nine testing events per isolate) with a common lot of microdilution trays. Agreement among MICs (i.e., mode +/- 1 twofold dilution) varied considerably for the different drug-isolate combinations and overall was best for cefoxitin (91.7 and 97.2% for one isolate each and 100% for all others), followed by doxycycline, amikacin, and ciprofloxacin. Agreement based on the interpretive category, using currently suggested breakpoints, also varied and overall was best for doxycycline (97.2% for one isolate and 100% for the rest), followed by ciprofloxacin and clarithromycin. Reproducibility among MICs and agreement by interpretive category was most variable for imipenem. Based on results reported from the individual sites, it appears that inexperience contributed significantly to the wide range of MICs of several drugs, especially clarithromycin, ciprofloxacin, and sulfamethoxazole. New interpretive guidelines are presented for the testing of M. fortuitum against clarithromycin; M. abscessus and M. chelonae against the aminoglycosides; and all three species against cefoxitin, doxycycline, and imipenem.

Amikacin↗

Influence of sodium fumarate addition on rumen fermentation in vitro.

The influence of sodium fumarate on rumen fermentation was investigated in vitro using batch and semi-continuous cultures of mixed rumen micro-organisms taken from three sheep receiving a basal diet of hay, barley, molasses, fish meal and a mineral-vitamin supplement (500, 299.5, 100, 91 and 9.5 g/kg DM respectively). Batch cultures consisted of 10 ml strained rumen fluid in 40 ml anaerobic buffer containing 200 mg of the same feed given to the sheep. Sodium fumarate was added to achieve a final concentration of 0, 5 or 10 mmol/l, as a result of the addition of 0, 250 or 500 mumol, equivalent to 0, 200 and 400 g/kg feed. CH4 production at 24 h (360 mumol in the control cultures) fell (P < 0.05) by 18 and 22 mumol respectively (SED 7.5). Total gas production was increased by the addition of fumarate without significant accumulation of H2. Substantial increases in acetate production (92 and 194 mumol; SED 26.7, P < 0.01) were accompanied by increases in propionate formation (212 and 396 mumol; SED 13.0, P < 0.001). Longer-term effects of fumarate supplementation on ruminal fermentation and CH4 production were investigated using the rumen simulation technique (Rusitec). Eight vessels were given 20 g basal diet/d, and half of them received a supplement of fumarate (disodium salt) over a period of 19 d. The response to the daily addition of 6.25 mmol sodium fumarate was a decrease in CH4 production of 1.2 mmol (SED 0.39, P < 0.05), equivalent to the consumption of 4.8 mmol H2, and an increase in propionate production of 4.9 mmol (from 10.4 to 15.3 (SED 1.05) mmol/d, P < 0.01). The inhibition of CH4 production did not decline during the period of time that fumarate was added to the vessels. Thus, the decrease in CH4 corresponded well to the fraction of the fumarate that was converted to propionate. Fumarate had no significant (P > 0.05) effect on total bacterial numbers or on the number of methanogenic archaea, but numbers of cellulolytic bacteria were increased (8.8 v. 23.9 (SED 2.49) x 10(5) per ml, P < 0.01). Fumarate also increased DM digestibility of the basal diet after 48 h incubation (0.476 v. 0.508 (SED 0.0123), P < 0.05). Thus, it was concluded that sodium fumarate may be a useful dietary additive for ruminants, because it diverts some H2 from CH4 production and because it is able to stimulate proliferation of cellulolytic bacteria and digestion of fibre.

Acetates↗

Influence of peptides and amino acids on fermentation rate and de novo synthesis of amino acids by mixed micro-organisms from the sheep rumen.

The influence of different N sources on fermentation rate and de novo amino acid synthesis by rumen micro-organisms was investigated in vitro using rumen fluid taken from four sheep receiving a mixed diet comprising (g/kg DM): grass hay 500, barley 299.5, molasses 100, fish meal 91, minerals and vitamins 9.5. Pancreatic casein hydrolysate (P; comprising mainly peptides with some free amino acids; 10 g/l), free amino acids (AA; casein acid hydrolysate + added cysteine and tryptophan; 10 g/l), or a mixture of L-proline, glycine, L-valine and L-threonine (M; 0.83 g/l each) were added to diluted (1:3, v/v), strained rumen fluid along with 15NH4Cl (A; 1.33 g/l) and 6.7 g/l of a mixture of starch, cellobiose and xylose (1:1:1, by weight). P and AA, but not M, stimulated net gas production after 4 and 8 h incubation (P < 0.05) in comparison with A alone. P increased microbial-protein synthesis (P < 0.05) compared with the other treatments. All of the microbial-N formed after 10 h was synthesized de novo from 15NH3 in treatment A, and the addition of pre-formed amino acids decreased the proportion to 0.37, 0.55, and 0.86 for P, AA, and M respectively. De novo synthesis of amino acids (0.29, 0.42 and 0.69 respectively) was lower than cell-N. Enrichment of alanine, glutamate and aspartate was slightly higher than that of other amino acids, while enrichment in proline was much lower, such that 0.83-0.95 of all proline incorporated into particulate matter was derived from pre-formed proline. Glycine, methionine, lysine, valine and threonine tended to be less enriched than other amino acids. The form in which the amino acids were supplied, as P or AA, had little influence on the pattern of de novo synthesis. When the concentration of peptides was decreased, the proportion of microbial-N formed from NH3 increased, so that at an initial concentration of 1 g peptides/l, similar to the highest reported ruminal peptide concentrations, 0.68 of cell-N was formed from NH3. Decreasing the NH3 concentration at 1.0 g peptides/l caused proportionate decreases in the fraction of cell-N derived from NH3, from 0.81 at 0.53 g NH3-N/l to 0.40 at 0.19 g NH3-N/l. It was concluded that different individual amino acids are synthesized de novo to different extents by mixed rumen micro-organisms when pre-formed amino acids are present, and that the source of N used for synthesis of cell-N and amino acids depends on the respective concentrations of the different N sources available; however, supplementing only with amino acids whose synthesis is lowest when pre-formed amino acids are present does not stimulate fermentation or microbial growth.

Amino Acids↗

Phylogenetic position and codon usage of two centrin genes from the rumen ciliate protozoan, Entodinium caudatum.

A lambda phage cDNA expression library was constructed from washed suspensions of the rumen ciliate protozoan, Entodinium caudatum, which had been maintained in an isolated, monofaunated sheep. The library was screened using an anti-E. caudatum antiserum raised in rabbits against sonically disrupted protozoa, DNA sequences for two centrins or caltractins, a subfamily of the EF-hand Ca(2+)-modulated proteins which are closely related, highly conserved cytoskeletal proteins, were identified and characterised. Their phylogenetic position was established relative to other centrin gene sequences. The two proteins showed homology to Paramecium tetraurelia centrins: 50 and 52% identities and 71 and 75% similarities in the protein sequence, over 99 and 110 amino acids fragments. Codon usage and indices revealed the E. caudatum follows universal codon usage, but with a restricted number of codons, and has a low G&C content.

Animals↗

Results of operation in Mycobacterium avium-intracellulare lung disease.

BACKGROUND: Although operation remains part of the management of Mycobacterium avium-intracellulare lung disease, few series have assessed operation in the era of better therapeutic drugs (especially clarithromycin). METHODS: From January 1, 1989, through June 30, 1997, 28 patients with M avirum-intracellulare lung disease underwent pulmonary resection. All were receiving multidrug therapy (17 of 28 were receiving clarithromycin) before and after operation. Eight patients underwent pneumonectomy (6 right, 2 left); 20 patients underwent partial resections including 18 with upper lobe lobectomies (14 right, 4 left). The most common indications for operation were medical treatment failure (15) and as part of initial therapy (9). RESULTS: Mean postoperative follow-up was 39 months. Complications occurred in 9 of 28 patients (32%), and included persistent air leak requiring surgical correction (5), early postoperative death (2), and late bronchopleural fistulae (1 patient). Twenty-three of 26 patients were known to be acid fast bacilli culture negative within 1 month of operation. Only 1 of 26 patients who survived 2 years is known to have had a relapse. CONCLUSIONS: Operation continues to play an important role in treatment of M avium-intracellulare lung disease. More than 90% of patients become culture negative and remain so when they continue to receive drugs. Although morbidity is relatively high, it is manageable and the 12-month mortality in the current series was low (7%).

Adult↗

Evaluation of a disk diffusion method for determining susceptibility of Mycobacterium avium complex to clarithromycin.

We evaluated an agar disk diffusion method for determining the susceptibility of Mycobacterium avium complex to clarithromycin. Isolates were inoculated onto the surface of a Middlebrook 7H11 plate, followed by the application of a 15-microgram clarithromycin disk. Zone sizes were read after 5-7 days of incubation. Zone sizes had a bimodal distribution; 40 isolates (10%) had no zone of inhibition, whereas the zone sizes for the remaining isolates ranged from 11 to 60 mm. Most isolates (37/40) having no zone of inhibition came from patients who had been treated previously with clarithromycin. Fifty-one isolates were also tested for clarithromycin susceptibility using a microdilution broth method. Defining susceptibility as a zone size of > 10 mm, disk diffusion test results agreed with the results by the microdilution broth method for 50 of 51 (98%) isolates tested by both methods. Agar disk diffusion is a promising method for the determination of clarithromycin susceptibility testing for M. avium complex.

Animals↗

AIDS-related Mycobacterium kansasii infection with initial resistance to clarithromycin.

Clarithromycin is a promising drug for the treatment of Mycobacterium kansasii infection. We report a patient with AIDS and severe M. kansasii infection who had previously received a short course of clarithromycin for sinusitis. He had clinical failure of treatment using clarithromycin plus ethambutol, and the initial isolate was found to be highly resistant to clarithromycin. Nucleotide sequencing of the 23S rRNA gene of this isolate demonstrated a single base mutation at position 2058, the same as that found in clarithromycin-resistant Mycobacterium avium.

AIDS-Related Opportunistic Infections↗

Uptake of acetylated peptides from the small intestine in sheep and their nutritive value in rats.

Acetylation is a potential method for protecting dietary peptides from degradation by rumen micro-organisms. As a first step in determining the nutritive value of acetylated peptides, their disappearance in the small intestine of sheep and their ability to support growth in a rat bioassay were measured. 15N-labelled peptides were prepared from lucerne which had been grown with 15N-labelled (NH4)2SO4 in the absence of Rhizobium. Peptides were prepared by enzymic hydrolysis of the extracted protein. Two peptide preparations were made using different proteinase mixtures. These mixtures contained peptides with an average molecular weight of 559 and 522 Da. They were treated with acetic anhydride, which resulted in 85 and 88% modification respectively, and their uptake from the small intestine was determined by injecting 1 g of untreated or acetylated peptides in a Cr-EDTA solution into the jejunum of two sheep fitted with jejunal catheters and ileal cannulas. Ileal digesta were collected and analysed for Cr and 15N. The uptake of dialanine (Ala2) and N-acetyl-Ala2 were compared in a similar way. The disappearance of 15N from lucerne peptides was high (88 and 93% respectively) and this was not affected significantly by acetylation (86 and 87%). Corresponding values for Ala2 and N-acetyl-Ala2 were both 96%, as measured by HPLC. It was therefore concluded that acetylation did not affect the uptake of peptides from the small intestine in sheep. Two feeding trials were carried out with rats. The first trial was carried out with a protein-free diet to which was added 10% lactalbumin or 5% lactalbumin and then a mixture of methionine-free amino acids, either alone or supplemented with Met, Gly-Met or acetylated Gly-Met. The rats grew equally well on all sources of Met, but failed to grow significantly on the mixture of Met-free amino acids. In the second trial the diet contained casein as 5.9% of the basal diet. Additional casein, pancreatic casein hydrolysate (peptides) and acetylated pancreatic casein hydrolysate (acetylated peptides) were compared as sources of amino acids, at inclusion rates of 100 g/kg final diet. Feed intake was similar with casein and peptides treatments, but was depressed by 23% with acetylated peptides. Live weight gain was 15 and 75% lower with the peptides and acetylated peptides diets respectively. Addition of lysine, arginine or histidine did not restore feed intake or weight gain of rats receiving acetylated peptides, but feed intake was restored immediately when peptides replaced acetylated peptides. When intake was restricted to 9 g/d and acetylated casein hydrolysate replaced half of the protein in the diet, rats gained weight less rapidly (1.44 v. 1.09 g/d) and retained less N, such that only 0.36 of the acetylated peptide-N was calculated to remain available to the animal. This N retention compared with 0.70 for unmodified casein. Thus, the rat bioassay indicated that certain specific peptides may well be of high nutritive value following acetylation, but that there may be problems of inappetance and inefficient utilization with acetylated peptide mixtures.

Acetylation↗

Variation in proteinase activities in the rumen.

Rumen fluid was taken from fistulated sheep and a cow receiving various diets based on grass hay or grass cubes with and without cereal-based concentrates. Proteinases in the extracellular fluid, and extracted from particulate material by Triton X-100, were visualized using SDS-PAGE in which gelatin was co-polymerized with the gels. Each animal sampled had a different pattern of proteinase activity. No single proteinase band predominated, although a few appeared in several samples, indicating that some microbial species were commonly involved in proteolysis but none was dominant. The banding patterns in samples taken from the same animals two weeks apart were fairly similar, indicating some stability within animals. Patterns obtained with extracellular fluid and Triton X100 extracts of small and large particulate material from the same sample were similar for the most part, although the relative intensity of the bands differed. The serine protease inhibitor, phenyl methyl sulphonyl fluoride, had little influence on the banding pattern. Concentrate in the diet appeared to increase inter-animal variation, and sheep in adjacent pens and consuming the same grass hay:concentrate diet had different banding patterns. Thus, no microbial proteolytic enzyme was predominant in protein digestion in the rumen.

Animal Feed↗

Influence of dietary acetylated peptides on fermentation and peptidase activities in the sheep rumen.

The predominant mechanism of peptide breakdown by rumen micro-organisms is aminopeptidase. Thus acetylation of the N-terminus of peptides inhibits their degradation by rumen micro-organisms in short-term incubations with rumen fluid in vitro. An experiment was undertaken to determine if adaptation of the rumen microbial population would take place when acetylated peptides were fed for a prolonged period, which would enable the microbial population to break down the protected peptides and thus decrease their nutritive value. Three adult sheep, fitted with permanent rumen cannulae, received a maintenance hay/concentrate diet to which was added, at each meal, 20 g of casein enzymic hydrolysate ('peptides') or 20 g of peptides previously treated with acetic anhydride. The diets were fed for 28 d in a 3 x 3 latin square and samples were taken during the last 7 d. Fermentation products and NH3 concentrations indicated that acetylated peptides remained less degradable than untreated peptides. There was a trend towards increased proteolytic activity with acetylated peptides, and dipeptidase activity increased by 18% and 28%, respectively, compared with untreated peptides and control treatments. Activity against N-acetyl-Ala2 also increased when acetylated peptides were fed, but it remained only 13% of the rate of Ala2 hydrolysis. No increase was found in the rate of ammonia production from acetylated peptides in animals receiving acetylated peptides--this rate was 26% of that found with untreated peptides--and acetylated peptides continued to persist for longer in the rumen than untreated peptides after feeding. Thus it was concluded that the rumen microbial population did not adapt to utilize acetylated peptides.

Acetylation↗

Long-term laboratory contamination by Mycobacterium abscessus resulting in two pseudo-outbreaks: recognition with use of random amplified polymorphic DNA (RAPD) polymerase chain reaction.

Beginning in 1993, an increase in clinical isolates of Mycobacterium abscessus was observed in a single hospital microbiology laboratory. This involved a cluster of four patients in June 1993 and five patients and a quality-control culture of distilled water in May 1994. Twenty-three M. abscessus isolates recovered between 1991 and 1996 were compared by random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). Sixteen of 21 clinical isolates recovered over a 6-year period and the distilled water isolate had identical RAPD-PCR patterns consistent with a single strain or clone. Only six of 15 patients had findings suggestive of clinical disease. Since the use of in-house-prepared distilled water was discontinued, no further laboratory contamination of clinical specimens has been observed. Molecular typing was the key to defining distilled water as the source of this pseudo-outbreak. Recognition of such outbreaks is important for prevention of unnecessary therapeutic and diagnostic interventions.

Adolescent↗

A single 16S ribosomal RNA substitution is responsible for resistance to amikacin and other 2-deoxystreptamine aminoglycosides in Mycobacterium abscessus and Mycobacterium chelonae.

Twenty-six clinical isolates of Mycobacterium abscessus resistant to amikacin were identified. Most isolates were from patients with posttympanostomy tube placement otitis media or patients with cystic fibrosis who had received aminoglycoside therapy. Isolates were highly resistant (MICs > 1024 microg/mL) to amikacin, kanamycin, gentamicin, tobramycin, and neomycin (all 2-deoxystreptamine aminoglycosides) but not to streptomycin. Sequencing of their 16S ribosomal (r) RNA revealed that 16 (94%) of 17 had an A-->G mutation at position 1408. In vitro-selected amikacin-resistant mutants of M. abscessus and Mycobacterium chelonae had the same resistance phenotype, and 15 mutants all had the same A-->G substitution at position 1408. Introducing an rRNA operon from Mycobacterium smegmatis with a mutated A-->G at this position into a single functional allelic rRNA mutant of M. smegmatis produced the same aminoglycoside resistance phenotype. These studies demonstrate this 16S rRNA mutation is responsible for amikacin resistance in M. abscessus, which has only one copy of the rRNA operon.

Alleles↗

Initial (6-month) results of three-times-weekly azithromycin in treatment regimens for Mycobacterium avium complex lung disease in human immunodeficiency virus-negative patients.

Two consecutive, open, prospective trials of intermittent azithromycin (600 mg), usually given Monday, Wednesday, and Friday (TIW) for Mycobacterium avium complex (MAC) lung disease were initiated in human immunodeficiency virus-negative patients. Regimen A consisted of TIW azithromycin and daily ethambutol (15 mg/kg/day), daily rifabutin (300 mg/day), and initial twice weekly (BIW) streptomycin. Regimen B consisted of TIW azithromycin, TIW ethambutol (25 mg/kg/dose), TIW rifabutin (600 mg/dose), and initial BIW streptomycin. Of 19 patients enrolled in regimen A who completed at least 6 months of therapy, 14 (74%) had sputum samples become culture-negative. Of 39 patients enrolled in regimen B who completed at least 6 months of therapy, 24 (62%) had sputum conversion. These sputum conversion rates are comparable to previous rates at 6 months in patients receiving daily clarithromycin- or azithromycin-containing regimens. No resistance to azithromycin emerged with either regimen. This is the first study to demonstrate the efficacy of intermittent administration of medication for MAC lung disease.

Anti-Bacterial Agents↗

De novo synthesis of amino acids by the ruminal bacteria Prevotella bryantii B14, Selenomonas ruminantium HD4, and Streptococcus bovis ES1.

The influence of peptides and amino acids on ammonia assimilation and de novo synthesis of amino acids by three predominant noncellulolytic species of ruminal bacteria, Prevotella bryantii B14, Selenomonas ruminantium HD4, and Streptococcus bovis ES1, was determined by growing these bacteria in media containing 15NH4Cl and various additions of pancreatic hydrolysates of casein (peptides) or amino acids. The proportion of cell N and amino acids formed de novo decreased as the concentration of peptides increased. At high concentrations of peptides (10 and 30 g/liter), the incorporation of ammonia accounted for less than 0.16 of bacterial amino acid N and less than 0.30 of total N. At 1 g/liter, which is more similar to peptide concentrations found in the rumen, 0.68, 0.87, and 0.46 of bacterial amino acid N and 0.83, 0.89, and 0.64 of total N were derived from ammonia by P. bryantii, S. ruminantium, and S. bovis, respectively. Concentration-dependent responses were also obtained with amino acids. No individual amino acid was exhausted in any incubation medium. For cultures of P. bryantii, peptides were incorporated and stimulated growth more effectively than amino acids, while cultures of the other species showed no preference for peptides or amino acids. Apparent growth yields increased by between 8 and 57%, depending on the species, when 1 g of peptides or amino acids per liter was added to the medium. Proline synthesis was greatly decreased when peptides or amino acids were added to the medium, while glutamate and aspartate were enriched to a greater extent than other amino acids under all conditions. Thus, the proportion of bacterial protein formed de novo in noncellulolytic ruminal bacteria varies according to species and the form and identity of the amino acid and in a concentration-dependent manner.

Amino Acids↗