Biomedical subjects
G Hughes
Publications and source records attributed to G Hughes.
Capsazepine: a competitive antagonist of the sensory neurone excitant capsaicin.
1. Capsazepine is a synthetic analogue of the sensory neurone excitotoxin, capsaicin. The present study shows the capsazepine acts as a competitive antagonist of capsaicin. 2. Capsazepine (10 microM) reversibly reduced or abolished the current response to capsaicin (500 nM) of voltage-clamped dorsal root ganglion (DRG) neurones from rats. In contrast, the responses to 50 microM gamma-aminobutyric acid (GABA) and 5 microM adenosine 5'-triphosphate (ATP) were unaffected. 3. The effects of capsazepine were examined quantitatively with radioactive ion flux experiments. Capsazepine inhibited the capsaicin (500 nM)-induced 45Ca2+ uptake in cultures of rat DRG neurones with an IC50 of 420 +/- 46 nM (mean +/- s.e.mean, n = 6). The 45Ca2+ uptake evoked by resiniferatoxin (RTX), a potent capsaicin-like agonist was also inhibited. (Log concentration)-effect curves for RTX (0.3 nM-1 microM) were shifted in a competitive manner by capsazepine. The Schild plot of the data had a slope of 1.08 +/- 0.15 (s.e.) and gave an apparent Kd estimate for capsazepine of 220 nM (95% confidence limits, 57-400 nM). 4. Capsazepine also inhibited the capsaicin- and RTX-evoked efflux of 86Rb+ from cultured DRG neurones. The inhibition appeared to be competitive and Schild plots yielded apparent Kd estimates of 148 nM (95% confidence limits, 30-332 nM) with capsaicin as the agonist and 107 nM (95% confidence limits, 49-162 nM) with RTX as agonist. 5. A similar competitive inhibition by capsazepine was seen for capsaicin-induced [14C]-guanidinium efflux from segments of adult rat vagus nerves (apparent Kd = 690 nM; 95% confidence limits, 63 nM-1.45 microM). No significant difference was noted in the apparent Kd estimates for capsazepine in assays on cultured DRG neurones and vagus nerve as shown by the overlap in the 95% confidence limits.6. Capsazepine, at concentrations up to 1O microM, had no significant effects on the efflux of 86Rb+ from cultured DRG neurones evoked either by depolarization with high (50 mM) K' solutions or by acidification of the external medium to pH 5.0-5.6. Similarly capsazepine had no significant effect on he depolarization (50 mM KCl)-induced efflux of [14C]-guanidinium from vagus nerve preparations.7. Ruthenium Red was also tested for antagonism against capsaicin evoked ['4C]-guanidinium release from vague nerves and capsaicin induced 45Ca2" uptake in cultures of DRG neurones. In contrast to capsazepine the inhibition by Ruthenium Red (10-500nM in DRG and 0.5-10microM in vagus nerve experiments) was not consistent with a competitive antagonism, but rather suggested a more complex,non-competitive inhibition.
Interview with Graham Hughes. What is going to happen tomorrow in the field of anti-phospholipid antibodies?. Interview by P. Youinou.
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The putative single-stranded DNA-binding protein of the filamentous bacteriophage, Ifl. Amino acid sequence of the protein and structure of the gene.
The protein product corresponding to the gene located in the region of the coliphage Ifl genome shown to contain the code for the single-stranded DNA (ssDNA)-binding proteins of all filamentous phages so far studied has been isolated from infected bacterial cells and its amino acid sequence determined. The mature protein contains 95 amino acids (calculated molecular mass 10553 Da). Its sequence corresponds to that predicted from the DNA sequence but lacks the initiating methionine residue. Although there is little direct sequence homology between the phage Ifl protein and the ssDNA-binding proteins of the other filamentous phages that have been studied, computer-based comparisons of various physical and structural parameters showed that the phage Ifl protein contains a domain that is closely related to domains in the coliphage T4 gene 32 protein and the Pseudomonas phage Pfl ssDNA-binding protein and suggest that the Ifl protein does have a ssDNA-binding function although we were unable to show this directly.
Evaluation of two endotracheal suction regimes in babies ventilated for respiratory distress syndrome.
All low birthweight babies ventilated for the respiratory distress syndrome during a period of fourteen months were randomised to receive endotracheal suction 12-hourly or 6-hourly. There were no significant differences in respiratory outcome between the groups. These observations suggest that it is safe to aspirate endotracheal tubes infrequently during the first few days in uncomplicated respiratory distress syndrome.
Pulmonary function and cardiovascular risk factor relationships in black and in white young men and women. The CARDIA Study.
Pulmonary function is known to be related inversely to incidence of coronary heart disease, congestive heart failure, chronic obstructive lung disease, lung cancer, and death from all causes. Reasons for some of these associations are poorly understood. Relationships between cardiovascular disease risk factors and pulmonary function were examined in 5,115 18- to 30-year-old black and white male and female participants in the study of Coronary Artery Risk Development in Young Adults (CARDIA). Forced expiratory volume in 1 s adjusted for height (FEV1/Ht2) was significantly lower in smokers than nonsmokers and in persons who reported shortness of breath; FEV1/Ht2 was correlated positively with a history of strenuous physical activity, duration of exercise on the treadmill, and high-density lipoprotein cholesterol. It was associated negatively with skinfold thicknesses, serum triglycerides, fasting serum insulin, and the Cook Medley scale of hostility. The association between pulmonary function and heart disease risk may reflect associations with physical fitness, vigor, fatness, and lipid profiles, as well as with cigarette smoking.
Families, parks, gardens and toxocariasis.
Toxocara canis antibody titres were measured in members of 30 families. 73/140 (52.1%) family members were seropositive for T. canis at a dilution greater than or equal to 1:50 and 11 (7.8%) had reciprocal titres greater than or equal to 800. Toxocara ova were sought (a) in relation to the 30 families and (b) in the community. No toxocara ova were found in household dust samples. 38% of family gardens contained toxocara ova while 6% of soil samples from public parks and open spaces around housing estates were positive. None of the faecal specimens from pet dogs were positive for toxocara ova as compared to 6.2% of stray dogs. 5.3% of canine faecal samples from the streets contained toxocara ova. In this study household garden soil was a potentially greater source of toxocara infection than soil in public parks and open spaces.
[Wegener's granulomatosis. New aspects of diagnosis, treatment and prognosis].
In its classical form, Wegener's granulomatosis (WG) is characterized by acute granulomatous vasculitis of the upper and lower respiratory tract together with glomerulonephritis, and when untreated, mortality is high. The use of cyclophosphamide and prednisolon has improved the prognosis dramatically, while azathioprine and trimetoprim-sulfamethoxazole are of limited therapeutic value in these patients. Only 1/3 of subacute and chronic cases show renal impairment, and the prognosis is better in limited Wegener's granulomatosis without renal disease than in the classical form. This case-report and review takes up various new aspects of diagnosis, treatment and prognosis.
Selection, training, and quality control of Type A interviewers in a prospective study of young adults.
This paper describes Type A/B interviewer selection, training, and quality control results in a prospective study of coronary artery risk development in young adults (CARDIA). Interviewer behaviors from 152 CARDIA structured interviews were audited and compared with 747 Western Collaborative Group Study (WCGS) interviews and 577 Multiple Risk Factor Intervention Trial (MRFIT) interviews. The results show success in modeling the CARDIA interviewer behaviors on those of the WCGS. CARDIA interviews were very similar to WCGS interviews for interview length, number of questions asked, and speed of speaking; they were similar to MRFIT interviews in latency of asking questions. CARDIA interviewer behaviors remained fairly consistent over the four time periods. Comparing the clinics, there were regional differences in latency of asking and speed of speaking, with the Southern clinic having a longer asking latency and speaking more slowly. There were differences between individual interviewers in most characteristics, particularly those that were more free to vary. The study provides procedures and guidelines designed to maintain quality control of the structured interview process.
The role of carbohydrate in the antigenic and immunogenic structure of bovine herpesvirus type 1 glycoproteins gI and gIV.
The role of carbohydrate in the antigenic and immunogenic structure of bovine herpesvirus type 1 (BHV-1) glycoproteins gI and gIV was investigated. Deglycosylated proteins induced a significantly lower antibody response in rabbits than native glycoproteins suggesting that the immunogenicity of several epitopes on gI and gIV is carbohydrate-dependent. Loss of carbohydrate from gI also resulted in a significantly decreased ability to induce a serum neutralizing antibody response to BHV-1, due to modifications in three distinct carbohydrate-containing continuous epitopes. Similarly, in vitro lysis of BHV-1-infected cells was significantly reduced when antibodies raised against deglycosylated gI were employed; this was attributed to changes in two of the three carbohydrate-dependent neutralizing epitopes on gI. The oligosaccharides may be directly involved as actual components of these continuous epitopes, rather than in stabilization of the conformation of the protein. In contrast, carbohydrate removal from gIV did not have a significant effect on the capacity to stimulate a neutralizing antibody response. Accordingly, none of the neutralizing epitopes on gIV appeared to be carbohydrate-dependent. Similarly, lysis of virus-infected cells was not significantly reduced when antibodies specific for deglycosylated rather than native gIV were used. In contrast to the humoral response, the delayed-type hypersensitivity response was stronger in rabbits immunized with deglycosylated proteins than in those inoculated with native glycoproteins gI or gIV. Consequently, the carbohydrates on gI and gIV may play a dual role in the host's immune recognition and response by contributing to certain epitopes, but masking others. The implications for the development of a subunit vaccine against BHV-1 are discussed.
Nerve-mediated contractions of sheep mesenteric lymph node capsules.
1. Isometric tension was recorded in vitro from strips cut from the capsules of mesenteric lymph nodes of sheep. 2. One minute periods of field stimulation at frequencies of 1, 2, 4, 8 and 16 Hz (pulse duration, 0.3 ms) elicited tonic contractions of increasing force and duration. The stimulus frequency-response relationship began to flatten out at frequencies greater than 4 Hz, where the response was already 72% of that at 16 Hz. 3. The response to field stimulation was abolished by tetrodotoxin (1 microM). 4. Phentolamine, rauwolscine and prazosin (all 1 microM) reduced the response to field stimulation, while desipramine (1 microM), potentiated it. 5. Atropine (1 microM) was without effect on the response. 6. These results suggest that sheep mesenteric lymph node capsules have a noradrenergic innervation which modulates their tone via an action on alpha-adrenoceptors.
Tape measure to aid prescription in paediatric resuscitation.
A tape measure, based on 50th centile weight for height and designed to permit easy drug dosage calculation, endotracheal tube size and DC cardioversion current dosages in childrens' emergencies, was tested for reliability by medical and nursing staff with varying paediatric experience. We found that the tape measure gave a reproducible estimate of weight and suggest that its use would facilitate decision making by inexperienced medical and nursing staff in paediatric resuscitation when there is no time to weigh the child.
Pea early browning virus RNA1 encodes four polypeptides including a putative zinc-finger protein.
We have determined the complete nucleotide sequence of RNA1 of the tobravirus pea early browning virus [PEBV] from an overlapping series of cDNA clones. The 7073 nucleotide sequence contains four open reading frames [ORFs]. The 5' proximal ORF encodes a 141K polypeptide, and readthrough of the opal [UGA] termination codon of this ORF would lead to the synthesis of a second, 201K polypeptide. Both of these polypeptides have extensive amino acid homology with the putative replicase proteins of tobacco rattle virus [TRV] and tobacco mosaic virus [TMV]. The third ORF encodes a 30K polypeptide which has homology with the TRV 29K and TMV 30K putative cell-to-cell spread proteins. The fourth, 3' proximal ORF encodes a 12K polypeptide which has extensive homology with the TRV 16K protein whose function is unknown. Examination of the amino acid sequences of the 12K and 16K gene products reveals in each the presence of two multiple-cysteine/histidine motifs, a finding which suggests that these proteins might have zinc and/or nucleic acid-binding properties.
Ethnic differences in blood pressure, pulse rate, and related characteristics in young adults. The CARDIA study.
This study examined ethnic differences in blood pressure and pulse rate in young adults to see whether the differences, if they exist, can be explained by differences in body mass index, lifestyle, psychological, and socioeconomic characteristics. Data used were from the baseline examination of the Coronary Artery Risk Development in (Young) Adults Study (CARDIA). CARDIA is a longitudinal study of lifestyle and evolution of cardiovascular disease risk factors in 5,116 young adults, black and white, men and women, aged 18-30 years, of varying socioeconomic status. Young black adults had higher mean systolic blood pressure and slightly higher mean diastolic blood pressure than young white adults. For both men and women, the blood pressure differences between blacks and whites tended to be greater for the age group 25-30 than for the age group 18-24 years. Among the variables studied, body mass index, duration of exercise on the treadmill, number of cigarettes smoked per day, and number of alcoholic drinks per week were consistently associated with blood pressure. The blood pressure differences were greatly reduced after adjusting for these variables. Black participants had lower mean pulse rate than white participants. The differences tended to be greater for the age group 18-24 than for the age group 25-30 years. Among the variables studied, only duration on treadmill and number of cigarettes smoked per day were consistently correlated with pulse rate. With adjustment for duration on treadmill, the differences in pulse rate increased. These results suggest that differences in ethnic pattern of blood pressures and pulse rate with age may be due in part to obesity, physical fitness, alcohol consumption, and cigarette smoking.
Functional and topographical analyses of epitopes on bovine herpesvirus type 1 glycoprotein IV.
Bovine herpesvirus type 1 (BHV-1) glycoprotein gIV was purified by affinity chromatography. Purified preparations showed two distinct components of 71 K and 140 K following electrophoresis in sodium dodecyl sulphate polyacrylamide gels. The polypeptides were separated, excised from the gel and used to immunize rabbits; the resulting antisera showed a high degree of cross reactivity indicating that these polypeptides represent monomeric and dimeric forms of the same glycoprotein. Purified gIV was also used to develop a gIV-specific panel of monoclonal antibodies. Neutralizing monoclonal antibodies directed against gIV were conjugated to horseradish peroxidase and subjected to competition binding assays by ELISA. Three distinct neutralizing antigenic domains on gIV were identified. Domain 1 comprised two overlapping epitopes, whereas domain 2 was represented by a single monoclonal antibody. The third antigenic domain was made up of a complex of four identical or overlapping epitopes designated 3a, b, c, and d. Evidence is presented suggesting that domain 1 of gIV may be involved in penetration of the virus into the cell.
Studies on the in vitro biological activities of recombinant bovine tumor necrosis factor (rBoTNF) alpha. I. Synergistic antiviral efficacy of rBoTNF alpha, recombinant bovine interferon gamma (rBoIFN gamma) and their combination.
The recent demonstration of the antiviral activity of recombinant human TNF has launched an interest in the use of TNF alpha for antiviral therapy in veterinary medicine. In the precent report, we demonstrate that pretreatment of bovine cells with rBoTNF alpha reduces the yield of bovine herpesvirus type-1 (BHV-1) from infected cells. Reduction in yield was similar to that observed in the presence of rBoIFN gamma. Similarly, rBoTNF alpha was able to protect bovine cells from virus-induced cytopathology. Enhanced antiviral activity was demonstrated when rBoTNF alpha was administered in combination with rBoIFN gamma. Studies on the induction of 2',5'-oligoadenylate synthetase (2'-5' AS) production by cytokine-treated cells showed that although rBoTNF alpha by itself did not induce 2'-5' AS activity it was capable of enhancing the production of 2'-5' AS by rBoIFN gamma-treated cells. Combination of these two cytokines was also evident in the inhibition of proliferation of treated cells. In contrast, the cytotoxic effect of rBoTNF alpha towards actinomycin D-treated cells was not affected by the combination of rBoTNF alpha with rBoIFN gamma.
The compartmental distribution of oestrogens and pregnancy specific beta1 glycoprotein.
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Studies on pregnancy-associated plasma protein A in the third trimester of pregnancy.
The plasma concentration of pregnancy-associated plasma protein A (PAPP-A) was measured in 34 women during the last 10 weeks of pregnancy. From 30 to 36 weeks the concentration of this protein increased steadily. Thereafter the concentration of PAPP-A rose more steeply, the highest amounts being found in early labour. The concentration of PAPP-A in peripheral venous blood and in the uterine vein was much the same. It was less in the retroplacental blood and a great deal less in the peritoneal fluid. The day-to-day variation was small; the coefficient of variation at 38 weeks was only 7.3 per cent. After delivery, the concentration of PAPP-A fell more slowly than other placental proteins and steroids, the average half-life being 51 hours. Although there is no doubt that PAPP-A is a product of the syncytiotrophoblast, our findings suggest that it is not simply secreted by the chorionic villi directly into the intervillous space but makes its way into the maternal circulation by a more circuitous route.