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Biomedical subjects

A Jenkins

Publications and source records attributed to A Jenkins.

At least 109 records · Page 6Linked to original sources

A thermal clearance probe for continuous monitoring of cerebral blood flow.

Thermal clearance has been used as a measure of tissue blood flow for over 60 years. We have developed a probe which can be used to monitor cerebral cortical thermal clearance in patients suffering from acute neurosurgical disorders. Despite its limitations of size, invasiveness, small sample volume, and difficult quantification it has proved a reliable method, showing a good correlation between measured thermal clearance and the onset of clinical signs of ischaemia.

Adult↗

Population sequence analysis of a simian immunodeficiency virus (32H reisolate of SIVmac251): a virus stock used for international vaccine studies.

The virus structural genes gag and env (both gp120 and gp41 regions) of the 32H isolate of SIVmac251 were amplified using the polymerase chain reaction (PCR). The proviral template used in the PCR was DNA isolated from cells used to prepare several experimental SIV vaccines, which have been tested in simians, and a standard challenge stock of virus, which has been used in international collaborative studies. The PCR products were cloned and the nucleotide sequences of several clones were determined for each gene. From a comparison of the sequences obtained the predominant amino acid sequences of gag and env were predicted and the degree of sequence heterogeneity was determined. Conserved and more variable regions of each protein were identified. The gp120 region of env was more heterogeneous than gag or the transmembrane protein of env (gp41). Within gp120, sequence variability was concentrated to specific regions equivalent to the V1, V2, and, to a lesser extent, the C1 regions identified for human immunodeficiency virus type 1 (HIV-1). In contrast the region equivalent to the hypervariable "V3-loop" of HIV-1 was highly conserved. The implications of the data is discussed in relation to the ability of this virus stock to prepare effective vaccines against SIV.

Amino Acid Sequence↗

Comparison of HPV detection in parallel biopsies and cervical scrapes by PCR.

PCR was used to detect HPV DNA in cervical scrapes of 23 women with cervical cancer. Compared with PCR-assisted HPV DNA detection in parallel biopsies, the sensitivity was 81% and the specificity 100%. We conclude that cervical scrapes can be used for detection of HPV DNA in women with cervical cancer.

Base Sequence↗

Transformational exchanges in the dihydropteroate synthase gene of Neisseria meningitidis: a novel mechanism for acquisition of sulfonamide resistance.

The nucleotide sequences of the chromosomal dihydropteroate synthase (dhps) genes in sulfonamide-susceptible and sulfonamide-resistant strains of Neisseria meningitidis of serogroups A, B and C were determined. The molecular weights and the amino acid sequences showed similarity to those of all other known dihydropteroate synthase polypeptides. Sequence comparison of the N. meningitidis dhps genes indicated horizontal transfer of DNA segments rather than point mutations as the cause for resistance in meningococci. The dhps genes in three of four sulfonamide-resistant meningococci contained identical central regions of 424 bp. Compared with the corresponding genes in susceptible strains, each central region included an insert of 6 bp. In one of the sulfonamide-resistant strains, the dhps gene was similar to the corresponding genes in the sensitive strains in its NH2-terminal and C-terminal parts. Its central region, however, was identical to the corresponding regions of two of the other resistant genes, and thus it could be seen as a hybrid dhps gene. Transformation experiments and mapping of transformed dhps genes indicated the existence of a novel mechanism for the dissemination of sulfonamide resistance in N. meningitidis. The origin of the resistance-mediating segment of the gene is unknown, but hybridization results showed the presence of homologous dhps genes in Neisseria gonorrhoeae and N. lactamica but not in N. subflava or Branhamella catarrhalis.

Amino Acid Sequence↗

Rapid diagnosis of meningococcal meningitis by polymerase chain reaction.

Rapid diagnosis of meningococcal disease followed by early treatment is essential. However, culture of blood or cerebrospinal fluid (CSF) may be unsuccessful because antibiotic treatment is often started before adequate specimens are collected, and because bacteria may die during transportation to the laboratory. We have used the polymerase chain reaction (PCR) to detect meningococcal DNA in a culture-negative CSF of a 15-year-old girl with meningococcal disease. Two oligonucleotides flanking the dihydropteroate synthase gene (dhps) of Neisseria meningitidis were used as primers. The PCR reaction is a rapid technique for the early detection of meningococcal meningitis, and also when culture is negative.

Adolescent↗

Specific types of colonic fermentation may raise low-density-lipoprotein-cholesterol concentrations.

To assess the effects of increased colonic fermentation on serum lipids, eight healthy volunteers were placed on two identical 2-wk metabolic diets, one of which was supplemented with lactulose (18-25 g/d). Lactulose raised day-long concentrations of breath hydrogen and serum glutamine as indicators of increased colonic fermentation by 78 +/- 13% (P less than 0.001) and 24.7 +/- 9.5% (P less than 0.05), respectively). Unexpectedly, however, fasting serum total and low-density-lipoprotein cholesterol and apolipoprotein B concentrations were higher at 2 wk by 8.9 +/- 1.5% (P less than 0.001), 10.9 +/- 2.2% (P less than 0.005), and 18.9 +/- 5.9% (P less than 0.02), respectively, compared with the control diet. With lactulose, mean free fatty acid concentrations were reduced over the day by 19.5 +/- 5.9% (P less than 0.02), with no change in mean day-long blood glucose, serum insulin, or C-peptide concentrations. We conclude that certain rapidly fermented substrates may raise rather than lower serum lipids, possibly through increasing the amount of acetate absorbed from the colon.

Acetates↗

T cell sensitization to factor VIII in haemophilia A?

Previous studies have indicated that factor VIII concentrates have a direct inhibitory effect on the proliferation of peripheral blood mononuclear cells (PBMC) obtained from normal volunteers. We have studied the effects of an intermediate and high purity factor VIII concentrate on haemophiliac PBMC. Intermediate purity factor VIII concentrate inhibited PBMC proliferation, the high purity factor VIII concentrate had no effect. The high purity factor VIII concentrate increased production of the lymphokine interleukin 2, suggesting the presence of a clone of T cells sensitized to factor VIII.

Albumins↗

Detection of genital papillomavirus types by polymerase chain reaction using common primers.

We describe the detection of eight genital human papillomavirus (HPV) types, including HPV16 and HPV18, by PCR amplification of a 323 base-pair region of the genome within the L1 open reading frame (ORF). The primer sequences are: TGYAAATATCCWGATTWTWT and GTATCWACMACAGTAACAAA. The method will detect purified HPV16 DNA down to a concentration of as little as a single molecule in 100 microliters. The method is also applicable to purified DNA and crude lysates from tumour biopsies. Typing of the PCR product can be achieved with specific oligonucleotide probes.

Base Sequence↗

DNA fingerprinting of isolates of Staphylococcus aureus from newborns and their contacts.

During a study on the epidemiology of Staphylococcus aureus colonization in newborns, mothers, and hospital staff, S. aureus was isolated from 536 of 1,945 specimens. Ninety-three isolates of S. aureus from the three groups of individuals were included in a study to evaluate the potential of DNA fingerprinting for strain differentiation. The 93 isolates were also phage typed and their plasmid profiles were analyzed. Cleavage of DNA with BamHI resulted in 13 different DNA restriction endonuclease band patterns (DNA REBPs), one of which consisted of eight isolates whose DNA was not cleaved with BamHI. The DNAs from these eight isolates were easily cleaved with HindIII. The different DNA REBPs were stable both during in vitro and in vivo growth and allowed strain differentiation within phage groups or types. We could not show any strong association between DNA REBP classes, phage types or groups, and plasmid profiles. Of the 93 isolates, 27 (29.0%) could not be phage typed and 12 (12.9%) lacked plasmids. We therefore conclude that DNA fingerprinting is a powerful tool, in addition to phage typing and plasmid profile analysis, for strain differentiation of S. aureus.

Adult↗

Clinical experience with pamidronate in the treatment of Paget's disease of bone.

Bisphosphonates have been shown to be effective in treating the increased bone turnover associated with Paget's disease of bone. In this study two groups of patients were treated with pamidronate by intravenous infusion. In group 1 (n = 15) 30 mg of pamidronate was given once a week for six weeks. A subgroup (group 1A, n = 6) of more severely affected patients (pretreatment serum alkaline phosphatase (ALP) greater than 1000 U/l, normal range 80-280 U/l) received a further 60 mg weekly for three weeks. Group 2 (n = 24) received 45 mg of pamidronate every three months for one year. In both groups the level of ALP in serum samples decreased steadily throughout the year. In group 1 the level decreased to a mean value of 230 U/l (95% confidence interval 188-281) and in group 2 to 297 U/l (227-389). Four of the six patients in group 1A achieved normal ALP, whereas ALP remained at an increased level in all of the 10 patients in group 2 whose pretreatment ALP was greater than 1000 U/l, suggesting that a dose-response effect exists. The lowest hydroxyproline to creatinine ratios (normal ratio less than 0.033) were observed at the end of treatment in group 1, with a mean ratio of 0.022 (range 0.015-0.033) and at three months after the start of treatment in group 2 with a mean ratio of 0.029 (range 0.022-0.037). There was a significant decrease in the turnover of bone, as measured by whole body retention of radiolabelled bisphosphonate, from a mean of 49.3 to 41.0% (p less than 0.01). These data confirm that pamidronate is effective in the management of Paget's disease of bone. For patients with levels of ALP in serum samples of up to four times above the upper limit of the normal reference range, an effective and convenient regimen is 45 mg every three months for one year. For patients with higher levels of ALP higher doses may be more effective.

Adult↗

Is colonic fermentation minimized by increasing meal frequency?

It has been suggested that, although increasing meal frequency has metabolic advantages in terms of carbohydrate and lipid metabolism, the efficiency of small intestinal absorption may be enhanced, thus reducing the extent of colonic fermentation. Since this may be undesirable, we have tested whether increasing meal frequency reduces the amount of carbohydrate fermented into the colon. Accordingly, seven healthy male volunteers took, in random order, two identical 2-week metabolic diets divided into either seventeen or three meals daily. On day 13 of each period measurements were made throughout the day of breath H2 and serum acetate, as markers of colonic fermentation. Mean levels of both breath H2 and acetate were similar on both diets, being lower on nibbling by 3.2 +/- 0.8 ppm and 25 +/- 9 mumols/l respectively (not significant). This study failed to demonstrate an effect of more efficient carbohydrate absorption with increased meal frequency.

Acetates↗

The production and purification of PCR-derived recombinant simian immunodeficiency virus p27 gag protein; its use in detecting serological and T-cell responses in macaques.

The polymerase chain reaction (PCR) was used to amplify a region of the gag gene, encompassing the core protein p27, from genomic DNA of cells infected with SIVmac251 (32H isolate). The 767 base pair PCR product was cloned into the bacteriophage M13 and fully sequenced before sub-cloning into the expression vector pUC19. The 30 kilodalton (kDa) fusion protein of lacZ-p27 was expressed as a soluble protein in E. coli JM101 cells and purified to greater than 90% purity by affinity chromatography. The affinity purified product was used in serological and T-cell assays to assess immune function in cynomolgus macaques immunised or challenged with immunodeficiency virus derived material. This reagent and accompanying methods provide valuable assays for monitoring the efficacy of vaccines for SIV as a model for human AIDS.

Animals↗

Cloning and characterization of a DNA fragment that confers sulfonamide resistance in a serogroup B, serotype 15 strain of Neisseria meningitidis.

By cloning studies and complementation experiments, the sulfonamide resistance gene of a serogroup B and serotype 15 (B:15) strain of Neisseria meningitidis was localized to a 1.2-kb chromosomal SspI fragment expressing a drug-resistant dihydropteroate synthase. The fragment hybridized to DNA from both resistant and susceptible strains, suggesting that the resistance gene is a variant of the normal gene for dihydropteroate synthase.

Cloning, Molecular↗

Experimental intracerebral haematoma: the role of blood constituents in early ischaemia.

In patients with intracerebral haematoma, ischaemic damage and final outcome are often more serious than the size of the lesion would suggest. The aetiology of the ischaemia in relation to space-occupying effects or specific factors present in blood is unclear. In a rat model of an intracerebral space-occupying lesion, the pathophysiological effects of a haematoma were compared with those of an equal volume of inert fluid (mock cerebrospinal fluid [CSF] or silicone oil). Cerebral blood flow was measured at 1 min by 14C iodoantipyrine autoradiography, and ischaemic cell damage was assessed by light microscopy at 4 h. In all animals, cerebral blood flow was reduced immediately adjacent to the lesion. In the group with a haematoma, blood flow was reduced (p less than 0.001) over a greater radius and also in the ipsilateral frontal and parietal cortex. Ischaemic damage was seen in animals lesioned with blood or oil of blood viscosity, but not in animals with CSF lesions. These data suggest that both tissue pressure and vasoactive substances are components of the immediate reduction in blood flow following intracranial haemorrhage. Tissue pressure may be the more important factor in later ischaemic neuronal damage.

Animals↗

Should we tell them when their blood pressure is up?

We carried out two studies to determine the effects of feedback on subsequent blood pressure and heart rate readings in subjects without significant cardiovascular abnormalities. In both studies the subjects were randomly assigned to be told that their blood pressure was normal or was high or to receive no feedback at all; 3 minutes later another reading was taken and correct feedback provided. Study 1 was done in 114 patients who attended a family practice teaching unit for an office visit; subjects taking cardioactive medication or with chronically elevated blood pressure (diastolic pressure more than 95 mm Hg) or known low pressure (diastolic pressure less than 60 mm Hg) were excluded. Half of the subjects received feedback from a nurse and the other half from a physician. We found no effect of type of feedback or type of practitioner on subsequent readings. No adaptation of diastolic blood pressure or heart rate took place, whereas a similar rest period in the laboratory consistently triggers cardiovascular adaptation. Given the field nature of the study it was not clear whether the intervention was not powerful or whether the practitioner-patient interactions diffused the effects of an otherwise powerful intervention. Therefore, a second study with the same design was carried out in a controlled laboratory setting with 61 university students who believed they were in the adaptation phase of an experimental stress protocol. The subjects did not interact with the experimenter, who provided only the initial feedback, via intercom. The findings replicated those of study 1: type of feedback had no significant effect on subsequent blood pressure levels, and all types of feedback prevented cardiovascular adaptation. We recommend that patients be allowed to rest alone for at least 10 minutes before blood pressure is measured. Our findings suggest that practitioners need not be concerned about telling normotensive or borderline hypertensive patients that their blood pressure is elevated.

Adult↗