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Biomedical subjects

R E Williams

Publications and source records attributed to R E Williams.

At least 37 records · Page 2Linked to original sources
PubMed2001

Turkish variant late infantile neuronal ceroid lipofuscinosis (CLN7) may be allelic to CLN8.

One variant form of late infantile neuronal ceroid lipofuscinosis (LINCL) is found predominantly within the Turkish population (CLN7). Exclusion mapping showed that CLN7 was not an allelic variant of known NCL loci (CLN1, CLN2, CLN3, CLN5 or CLN6). Using the method of homozygosity mapping, a genome-wide search was undertaken and a total of 358 microsatellite markers were typed at an average distance of about 10 cM. A region of shared homozygosity was identified on chromosome 8p23. This telomeric region contained the recently identified CLN8 gene. A missense mutation in CLN8 causes progressive epilepsy with mental retardation (EPMR) or Northern epilepsy, which has so far been reported only from Finland and is now classified as an NCL. The mouse model mnd has been shown to carry a 1 bp insertion in the orthologous Cln8 gene. Statistically significant evidence for linkage was obtained in this region, with LOD scores > 3, assuming either homogeneity or heterogeneity. Flanking recombinants defined a critical region of 14 cM between D8S504 and D8S1458 which encompasses CLN8. This suggests that Turkish variant LINCL, despite having an earlier onset and more severe phenotype, may be an allelic variant of Northern epilepsy. However mutation analysis has not so far identified a disease causing mutation within the coding or non-coding exons of CLN8 in the families. The Turkish variant LINCL disease-causing mutation remains to be delineated.

Alleles↗
PubMed2001

Analysis of candidate genes in the CLN6 critical region using in silico cloning.

CLN6, the gene for variant late infantile neuronal ceroid lipofuscinosis, was mapped to a 4 cM region on chromosome 15q22-23. Subsequently the critical region was narrowed to less than 1 cM between microsatellite markers D15S988 and D15S1000 by additional marker typing in an expanded family resource. A physical map was constructed across this region using YAC and PAC clones and sequence was generated from two PAC clones. This sequence was analysed together with overlapping sequence generated by the Human Genome Project to identify genes within the region using an in silico cloning approach. In all, 29 genes have been identified and 18 have been analysed for mutations by direct sequencing. This powerful new approach will lead to the identification of CLN6.

Chromosome Mapping↗
PubMed2001

Gluteus medius and rump fat depths as additional live animal ultrasound measurements for predicting retail product and trimmable fat in beef carcasses.

This study was conducted to determine the ability of additional ultrasound measures to enhance the prediction accuracy of retail product and trimmable fat yields based on weight and percentage. Thirty-two Hereford-sired steers were ultrasonically measured for 12th-rib fat thickness, longissimus muscle area, rump fat thickness, and gluteus medius depth immediately before slaughter. Chilled carcasses were evaluated for USDA yield grade factors and then fabricated into closely trimmed, boneless subprimals with 0.32 cm s.c. fat. The kilogram weight of end-point product included the weight of trimmed, boneless subprimals plus lean trim weights, chemically adjusted to 20% fat, whereas the fat included the weight of trimmed fat plus the weight of fat in the lean trim. Prediction equations for carcass yield end points were developed using live animal or carcass measurements, and live animal equations were developed including ultrasound ribeye area or using only linear measurements. Multiple regression equations, with and without ultrasound rump fat thickness and gluteus medius depth, had similar R2 values when predicting kilograms of product and percentages of product, suggesting that these alternative variables explained little additional variation. Final unshrunk weight and ultrasound 12th-rib fat thickness explained most of the variation when predicting kilograms of fat. Rump fat and gluteus medius depth accounted for an additional 10% of the variation in kilograms of fat, compared with the equation containing final weight, ultrasound ribeye area, and ultrasound 12th-rib fat thickness; however, the two equations were not significantly different. Prediction equations for the cutability end points had similar R2 values whether live animal ultrasound measurements or actual carcass measurements were used. However, when ultrasound ribeye area was excluded from live animal predictions, lower R2 values were obtained for kilograms of product (0.81 vs 0.67) and percentages of product (0.41 vs 0.17). Conversely, the exclusion of ultrasound ribeye area had little effect on the prediction accuracy for kilograms of fat (0.75 vs 0.74) and percentage fat (0.50 vs 0.40). These data substantiate the ability of live animal ultrasound measures to accurately assess beef carcass composition and suggest that the alternative ultrasound measures, rump fat and gluteus medius depth, improve the accuracy of predicting fat-based carcass yields.

Adipose Tissue↗
PubMed2001

Tail-docking alters fly numbers, fly-avoidance behaviors, and cleanliness, but not physiological measures.

Tail docking is an animal well-being issue not only regarding the docking procedures but also because of concerns during fly season. To address the latter question, we selected eight cows that had been tail-docked in a previous experiment and eight nondocked cows matched by stage of lactation. Physiological, immunological, and behavioral measures were used to evaluate the well being of those cows housed in a tie-stall barn during fly season for 5 consecutive days. Behavior was observed for 5-min interval instantaneous scan samples for 1 h each at 0800, 1200, and 1600 h. Flies were counted before behavior observations. Blood samples were taken daily for plasma and leukocyte separation. Cows were scored on d 5 for cleanliness on a five-point scale. Docked cows were cleaner, but fly counts of docked cows were greater for total fly counts and rear leg counts. However, counts were not different on front legs. Time of day was significant, so each time of day was analyzed separately. Docked cows were observed to exhibit fewer tail swings at 0800 h, but docked cows tended to ruminate more at that time. Docked cows tended to stand less at the 1200 h observation. Total fly-avoidance behaviors were greater for all cows at the 1600-h observation. Only tail swings tended to be more frequent with docked cows, but foot stomps occurred only in the docked cows. Lymphocyte phenotypes, acute-phase proteins, and immunoglobulin concentrations did not differ. In conclusion, although docked cows were cleaner, as the fly numbers increase throughout the day, fly-avoidance behaviors also increased and foot stomping appeared as an alternative method for fly avoidance by docked cows.

Acute-Phase Proteins↗
PubMed2000

Cloning, sequencing and expression in Escherichia coli of the primary alcohol dehydrogenase gene from Thermoanaerobacter ethanolicus JW200.

The structural gene, adhA, for a thermostable primary alcohol dehydrogenase was cloned from Thermoanaerobacter ethanolicus JW200. Constitutive expression from its own promoter was observed in Escherichia coli. The nucleotide sequence of adhA corresponded to an open reading frame of 1197 bp, encoding a polypeptide of 399 amino acids with a calculated Mr of 43 192. Amino acid sequence analysis showed 67-69% identity with alcohol dehydrogenases from two archaeal species and 29-37% identity with bacterial type III alcohol dehydrogenases. This represents the first reported cloning of an alcohol dehydrogenase from a bacterial species that is both thermostable and active against primary long-chain alcohols.

Alcohol Dehydrogenase↗
PubMed2000

The structure of arachno-[B6H11]-, at -25 degrees C in (CD3)2O, is resolved via the ab initio/IGLO-GIAO/NMR procedure.

The arachno-[B6H11]- solution structure at -25 degrees C was clarified as fluxional compound 2 by applying the ab initio/IGLO/NMR method. The anion 2 can be derived from arachno-B6H12, 1, by the removal of the B2/B3 bridging hydrogen (2). No minimum on the potential energy surface could be found for an asymmetric complex, a, between [B5H8]- and BH3, which had been proposed originally. A Cs-symmetric [mu-(BH3)B5H8]- complex, A, only 3.2 kcal mol-1 higher in energy than 2, is the intermediate in the fluxional rearrangement observed on the NMR time scale. The transition structure [D] connecting 2 (Erel = 0.0) and A (Erel = 3.2) has a relative energy of 9.7 kcal mol-1. The elimination of both a and A as "most stable structure" candidates of arachno-[B6H11]- reinforces the early geometrical bonding systematics for boranes and carboranes.

Journal Article↗
PubMed2000

Inclusion of S-sepharose beads in the culture medium significantly improves recovery of secreted rBPI(21) from transfected CHO-K1 cells.

rBPI(23), a recombinant N-terminal fragment of human bactericidal/permeability-increasing protein (BPI), kills gram-negative bacteria and binds endotoxin. rBPI(21), a variant, in which cysteine 132 is changed to alanine, retains the activities of rBPI(23). Initial attempts using conventional ion-exchange chromatography to purify rBPI(23) from culture supernatants of transfected CHO-K1 cells resulted in lower than expected yields. Also, ELISA of supernatants from CHO-K1 transfectants expressing rBPI(23) or rBPI(21) yielded variable signals. Results from pulse-chase experiments using [(35)S]methionine had indicated that rBPI(23) could not be detected in the culture medium by 7 h of chase, suggesting that these proteins were degraded and/or bound to cells, media components, or vessel surfaces. To address these issues, we developed a novel process whereby sterile S-Sepharose beads were added directly to the cell culture medium. For attached cells, the beads were added to confluent cultures with serum-free medium for the expression phase, while for suspension-adapted cells, beads were added at the beginning of culture growth. The S-Sepharose was then separated from cells and media and washed, and BPI was eluted with high-salt buffer. This approach yielded up to a 50-fold improvement in recovery of rBPI(23) and rBPI(21) from roller bottles, shake flasks, and 2-liter fermenters. It also resulted in improved detection and quantitation of secreted rBPI(23) and rBPI(21) by ELISA. Results of competition binding studies with iodinated rBPI(21) in conjunction with unlabeled rBPI(21) and rBPI(23) or with heparin demonstrated that these proteins bound specifically and with high affinity to heparan-containing sites on the surface of the CHO-K1 cells. We conclude that the S-Sepharose included in the culture medium captures the BPI protein products as they are secreted and protects them from degradation and/or irreversible binding to cell surfaces. This method has been scaled up to a manufacturing process in large (2750 liter) fermenters for pharmaceutical production.

Animals↗
PubMed2000

A prospective study of faecal bile acids and colorectal cancer.

A prospective study of 7079 people aged 45-74 recruited through general practices in South Wales, Herefordshire and Edinburgh, Scotland was undertaken to test the hypothesis that faecal bile acids are implicated in the causation of large bowel cancer. The population was recruited between 1974 and 1980 and the response rate for stool collection was 67%. Bile acid analyses were performed on those cases that presented by 1990. It was decided in advance to examine the hypothesis separately for left- and right-sided bowel cancer because of known epidemiological differences between the two sites and to exclude the cases presenting within 2 years of the stool sample from the analyses because the cancer could have been present at recruitment and might have possibly affected faecal bile acid concentrations. Each case (n = 51 left-sided and 8 right-sided) was matched with three controls by age (within 5 years), sex, place of residence and time of providing the stool sample (within 3 months). Statistical analyses using conditional logistic regression showed no significant differences between the left-sided cases and controls for any of the concentrations of individual bile acids, total bile acid concentrations, faecal neutral steroids, percentage bacterial conversion and the ratio of lithocholic acid to deoxycholic acid concentrations. There was a statistically significant (P = 0.021) association of the presence of chenodeoxycholic acid (5/8 samples) in the right-sided cases compared with the controls (3/23), odds ratio 6.26 (95% confidence interval 1.19, 32.84). A high proportion of primary bile acids has also been found in other studies of patients with a genetic predisposition to proximal bowel cancer, however this pattern may also occur in low risk groups, such as Indian vegetarians, suggesting that they may predispose to right-sided bowel cancer only in the presence of other, as yet unknown factors. If bile acids are involved in the causation of large bowel cancer, they may be part of a complex set of interacting factors.

Aged↗
PubMed1999

A new locus for variant late infantile neuronal ceroid lipofuscinosis-CLN7.

To date two genes are known to be involved in variant LINCL, CLN5 and CLN6, which map to chromosomes 13q21 and 15q21-23. A subset of Turkish families with a variant phenotype has been identified. Affected individuals have curvilinear bodies and fingerprint profiles on EM but are recombinant at CLN5 and CLN6. These families appear to represent a new locus. Homozygosity mapping is being used to map this locus, which has been designated CLN7.

DNA Mutational Analysis↗
PubMed1999

Genetic and physical mapping of the CLN6 gene on chromosome 15q21-23.

CLN6, the gene for a variant late infantile neuronal ceroid lipofuscinosis, has been mapped to chromosome 15q21-23 by homozygosity mapping. At present the family resource consists of 31 families. By the analysis of additional polymorphic markers in this resource the critical region has been narrowed down from 12 cM to less than 4 cM. A physical map is being constructed using YAC and PAC clones as a prerequisite to transcript mapping.

Alleles↗
PubMed1999

Ultrastructural and electrophysiological correlation of the genotypes of NCL.

Several genetically different, but clinically similar childhood forms of neuronal ceroid lipofuscinosis are now recognized. Accurate diagnosis is important so that appropriate genetic advice can be given, molecular analysis can be undertaken, and prenatal testing can be considered. A combined clinical, electrophysiological, and histological (light and electron microscopy) approach offers the most reliable means of diagnosis in the majority of patients. Patients with an unusual presentation will also be identified.

Electroencephalography↗
PubMed1999

The European Concerted Action NCL Clinical Case Registry.

A European NCL Clinical Case Registry has been set up in London, as part of a European Union-funded Concerted Action project. Concerted Action participants provide written information about children which is then anonymized and entered on the Registry. Contributors are able to request information contained within the Registry for the purpose of epidemiological, molecular, diagnostic, or therapeutic research. Up to May 1998, 60 cases were entered on the Registry.

Europe↗
PubMed1999

Biochemical and neurotoxicological effects of L-2-chloropropionic acid on rodent brain.

L-2-Chloropropionic acid (L-CPA) is selectively toxic to cerebellar granule cells; necrosis is first observed in rats 36 h after L-CPA administration (750 mg/kg p.o.) and becomes marked by 48 h. L-CPA has also been shown to activate the mitochondrial pyruvate dehydrogenase (PDH) complex in fasted adult rats, resulting in reduced blood glucose and lactate levels. This study aimed to investigate the biochemical and neurotoxicological effects of L-CPA on the brain. Extracts, prepared from guinea-pig cerebellar and cerebral cortex slices incubated in the presence of L-CPA, were analysed using 1H magnetic resonance spectroscopy, 31P magnetic resonance spectroscopy, and amino acid analysis. Glucose metabolism was studied by monitoring the metabolism of [1-(13)C]glucose using gas chromatography/mass spectrometry. Increased glucose metabolism and decreases in the pool sizes of lactate and alanine were observed in both tissues, demonstrating activation of the PDH complex. Extracts were also prepared from the forebrain and cerebellum of animals that had been treated in vivo with L-CPA and analysed as described for the in vitro studies. Similar evidence for PDH activation was demonstrated at 2 and 24 h after dosing in both tissues. At 48 h after dosing, when signs of toxicity are observed, an increase in the lactate concentration and a decrease in N-acetylaspartate in the cerebellum but not in the forebrain confirmed the selective neurotoxic action of L-CPA. These results suggest that activation of the PDH complex does not directly lead to the delayed selective neurotoxicity of L-CPA.

Alanine↗
PubMed1999

Bactericidal/permeability-increasing protein inhibits growth of a strain of Acholeplasma laidlawii and L forms of the gram-positive bacteria Staphylococcus aureus and Streptococcus pyogenes.

Bactericidal/permeability-increasing protein (BPI) inhibited growth of cell wall-deficient Acholeplasma laidlawii and L forms of certain strains of Staphylococcus aureus and Streptococcus pyogenes. However, the same strains of S. aureus and S. pyogenes with intact cell walls were not susceptible to the growth-inhibitory effects of BPI.

Acholeplasma laidlawii↗
PubMed1999

Individual differences in physiological responses and type A behavior pattern.

The relationships between individual differences in psychophysiological responses and tendency of Type A behavior pattern (TABP) were investigated during mental arithmetic (MA) at a steady rhythm, challenging calculation (Uchida-Kraepelin serial addition test: UK test), music listening, and exposure to an 80 dB SPL of white noise. Each mental task was sustained for 5 minutes. Sixteen healthy Japanese adults, (10 males and 6 females) with an age from 18 to 36 years old volunteered for this study. The KG's Daily Life Questionnaire (KG Questionnaire) was used to investigate the tendency toward TABP, which included three sub-factors: aggression-hostility, hard-driving and time urgency, and speed-power items. Recorded physiological variables were respiratory rate (RR), skin resistance response (SRR), eyeblinks, and heart rate (HR) calculated using frequency analysis to render high frequency power (HF) and the ratio of low/high frequencies (L/H ratio). During the MA and UK tests, significant increases in HR, L/H ratio, RR occurred, while significant decreases in HF were observed. Eyeblinks significantly increased during the MA test and significantly decreased during the UK test. During music and white noise, no significant changes occurred except for SRR, which decreased significantly. The coefficient of variances in each response was over 20% for almost all variables, indicating that individual differences in the magnitude of each response were large, even if the direction (increase or decrease) of the change was the same in almost all subjects. The highest correlation coefficients (r) between the mean values of relative magnitude for each variable and TABP scores during the MA and UK tests were obtained for the L/H ratio (MA: r = 0.591, UK test: r = 0.577) and the RR (MA: r = -0.576, UK test: r = -0.511). These values were statistically significant (p < 0.05). Similar results were obtained for TABP sub-factors. Though other investigations have reported relationships between HF and TABP, we found no significant relationship. It was suggested that sympathetic nerve activity became greater for TABP individuals than for Type B individuals under stress conditions.

Adolescent↗
PubMed1998

Stem cell factor influences mast cell mediator release in response to eosinophil-derived granule major basic protein.

Stem cell factor (SCF) is an important mast cell growth, differentiation, and survival factor. We investigated whether SCF influenced the response of mouse mast cells to an IgE-independent stimulus, eosinophil-derived granule major basic protein (MBP). Mouse bone marrow cultured mast cells (BMCMC) were derived in either concanavalin-stimulated mouse spleen conditioned medium (CM) or SCF. The cloned growth, factor-independent mast cell line Cl.MC/C57.1 was also studied. BMCMC in SCF exhibited cytochemical staining properties, protease and histamine content, and increased serotonin uptake consistent with more mature differentiated mast cells as compared with BMCMC in CM or Cl.MC/ C57.1 cells. BMCMC in SCF released serotonin, 14C-labeled arachidonic acid metabolites and tumor necrosis factor-alpha (TNF-alpha) on stimulation with MBP, while no response was seen from either BMCMC in CM or Cl.MC/C57.1 cells. All three mast cell populations released mediators on stimulation with the cationic MBP analog, poly-L-arginine, indicating that the cationic charge did not explain the selective response of BMCMC in SCF to eosinophil-derived granule MBP. These findings show that SCF significantly influences mast cell differentiation and the responsiveness of mast cells to eosinophil-derived granule MBP.

Animals↗