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Biomedical subjects

S Patel

Publications and source records attributed to S Patel.

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

Large scale production of cyclohexanone monooxygenase from Escherichia coli TOP10 pQR239.

The cyclohexanone monooxygenase (CHMO) from Acinetobacter calcoaceticus NCIMB 9871 has been cloned into Escherichia coli in an L-arabinose inducible vector. The recombinant E. coli containing the L-arabinose inducible CHMO was grown at 1.5 litres under controlled conditions to determine the parameters for growth and induction. It was found that induction with 0.1% (w/v) L-arabinose at late logarithmic phase of growth and growth for a further 2.5 to 3 h gave the optimal CHMO titre ( approximately 3500 U.l(-1,) 630 U. g dry cell weight(-1)). High dissolved oxygen concentrations were shown to be deleterious to the CHMO titre. This influenced the strategy for growth and induction, and was optimal when the oxygen uptake rate was maximized but the dissolved oxygen concentration was zero. Finally, a 300 litre scale fermentation was carried out giving a total CHMO titre of >8 x 10(5) U.

Journal Article↗
PubMed2001

Comparison of diameter and perimeter methods for tumor volume calculation.

PURPOSE: Lesion volume is often used as an end point in clinical trials of oncology therapy. We sought to compare the common method of using orthogonal diameters to estimate lesion volume (the diameter method) with a computer-assisted planimetric technique (the perimeter method). METHODS: Radiologists reviewed 825 magnetic resonance imaging studies from 219 patients with glioblastoma multiforme. Each study had lesion volume independently estimated via the diameter and perimeter methods. Cystic areas were subtracted out or excluded from the outlined lesion. Inter- and intrareader variability was measured by using multiple readings on 48 cases. Where serial studies were available in noncystic cases, a mock response analysis was used. RESULTS: The perimeter method had a reduced interreader and intrareader variability compared with the diameter method (using SD of differences): intrareader, 1.76 mL v 7.38 mL (P < .001); interreader, 2.51 mL v 9.07 mL (P < .001) for perimeter and diameter results, respectively. Of the 121 noncystic cases, 23 had serial data. In six (26.1%) of those 23, a classification difference occurred when the perimeter method was used versus the diameter method. CONCLUSION: Variability of measurements was reduced with the computer-assisted perimeter method compared with the diameter method, which suggests that changes in volume can be detected more accurately with the perimeter method. The differences between these techniques seem large enough to have an impact on grading the response to therapy.

Brain↗
PubMed2001

Functional MRI of human auditory cortex using block and event-related designs.

fMRI of human auditory cortex response to sinusoidal tones of 200, 1000, and 3000 Hz was evaluated using block design and conventional and "silent" event-related designs. Conventional event-related fMRI revealed the timecourse of the BOLD response (approximately 5 sec to peak, approximately 4 sec full-width-half-max, and approximately 14 sec recovery to baseline). Both event-related, but not block, designs provided evidence for tonotopic organization in auditory cortex. Sources of low-frequency activation were more lateral and anterior than the sources of high-frequency activation (P < or = 0.05). In the block designs, repeated rapid stimulus presentation and the co-incidence of scanner noise preclude definition of tonotopic organization revealed in event-related approaches. Magn Reson Med 45:254-260, 2001.

Acoustic Stimulation↗
PubMed2001

Increased matrix metalloproteinase expression and activation following experimental acute pancreatitis.

BACKGROUND: The observation that matrix metalloproteinases (MMPs) are central to tissue remodeling and may contribute to organ failure prompted us to investigate the role of MMPs in acute pancreatitis. We hypothesize that increased expression and activation of MMP-2 and MMP-9 will correlate with organ injury following acute pancreatitis. METHODS: Acute pancreatitis was induced in five male rats by retrograde infusion of 5% sodium taurocholate into the pancreatic duct. Sham laparotomy was performed on five rats serving as a control. Pancreatitis was confirmed by histology and serum amylase levels. MMP-2 and MMP-9 activity and expression were assayed by gelatin zymography in the lungs and ascitic fluid of each animal. Lung permeability was assayed by Evans blue dye extravasation. Lung activity of MMP-2 and MMP-9 was confirmed by a specific fluorogenic MMP substrate assay. RESULTS: Lung permeability increased twofold in the animals with severe pancreatitis compared with sham. Analysis of the zymograms from lung homogenate revealed a threefold increase in active MMP-2 in severe pancreatitis compared with sham and no change in MMP-9 activity. Gelatin zymograms of peritoneal fluid from severe pancreatitis animals demonstrated increased levels of active MMP-2 and MMP-9 compared with the sham group. Increases in MMP activity were confirmed by MMP activity assay using a fluorogenic substrate. CONCLUSIONS: This study demonstrates a correlation between severity of acute pancreatitis and active MMP-2 and MMP-9 levels in the peritoneal fluid and MMP-2 activity in lung homogenate. The MMP-mediated degradation of the basement membrane offers a potential pharmacologic and therapeutic target for halting the final biologic outcome of severe pancreatitis.

Acute Disease↗
PubMed2001

Association between integrin-dependent migration capacity of neural stem cells in vitro and anatomical repair following transplantation.

In previous transplantation studies using neural stem cell lines immortalized by the temperature-sensitive SV40 large T-antigen, we have shown that animals with experimental hippocampal lesions resulting from four vessel occlusion recover spatial memory functions more effectively when grafted with the MHP36 cell line than with the MHP15 cell line [Gray et al. (1999). Philos. Trans. R. Soc. London Biol. Sci. 354:1407-1421]. In the present study, we have investigated the cellular and molecular basis of these differences in repair capacity both in vivo and in vitro. Using the same model of hippocampal damage we have shown that following transplantation MHP36 cells migrate and align within the damaged CA1 of the ipsilateral hippocampus. MHP15 cells, in contrast, migrate in a more indiscriminate pattern that does not reflect the anatomy of the region. To analyze the migratory properties of these two cell lines in more detail, we performed migration assays at a nonpermissive temperature on the extracellular matrix substrates laminin, fibronectin, and vitronectin. These showed that MHP36 cells have a greater migration potential than the MHP15 cells. While the pattern of cell surface extracellular matrix receptors of the integrin family was identical in both cell lines, the different degrees of migration on vitronectin were both blocked by inhibitors of alphaV integrins. Differences in integrin signaling therefore contribute to the greater migration potential of the repairing MHP36 cell line.

Animals↗
PubMed2001

Mathematical analysis of hemoglobin spectrophotometry in microvessels.

Spectrophotometry of hemoglobin in microvessels is commonly performed by collecting light either from a small region around the vessel centerline or from the entire lumen of the vessel. In the latter instance, parallel rays of light may not encounter the same amount of absorbing species. Hence, a phenomenon similar to the sieve effect reported in the literature on hemoglobin spectrophotometry may be expected to occur. Although it has been observed that under such circumstances nonlinearities in calibration characteristics arise, the implications of this effect on the interpretation of the spectrophotometric mean concentration have never been addressed so far. Mathematical analysis of hemoglobin spectrophotometry in microvessels, performed in this study, reveals that for practical situations the calibration curve is indeed nonlinear. Moreover, the spectrophotometric mean oxygen saturation is an overestimate of the mean oxygen saturation during oxygenation and an underestimate of the mean oxygen saturation during deoxygenation. These deviations depend upon the manner in which the total heme concentration is distributed within the lumen. Application of the analysis to artificial microvessels showed that the observed superior oxygen transport characteristics of flowing erythrocyte suspensions and hemoglobin solution mixtures could in part be due to the assumptions underlying the procedure used to interpret the experimental results. The implications of this result on models for oxygen transport in microvessels are discussed along with possible resolutions.

Calibration↗
PubMed2001

Sequence analysis of the plasmid pRRI2 from the rumen bacterium Prevotella ruminicola 223/M2/7 and the use of pRRI2 in Prevotella/Bacteroides Shuttle Vectors.

pRRI2 is a small cryptic plasmid from the rumen bacterium Prevotella ruminicola 223/M2/7 which has been used for the construction of shuttle vectors (pRH3 and pRRI207) that replicate in many Bacteroides/Prevotella strains as well as in Escherichia coli. Sequence analysis of pRRI2 reveals that it is a 3240-bp plasmid carrying two clear open reading frames. Rep, encoded by ORF1, shows 48 and 47% amino acid sequence identity with RepA proteins from Bacteroides vulgatus and Bacteroides fragilis, respectively. ORF2, named Pre, shares 34% amino acid sequence identity with a putative plasmid recombination protein from the Flavobacterium spp. plasmid pFL1 and 30% amino acid sequence identity with BmpH from B. fragilis Tn5520. Disruption of ORF1 with HindIII prevents replication and maintenance in Bacteroides spp. hosts, but shuttle vectors carrying pRRI2 interrupted within ORF2, by EcoRI*, are able to replicate. pRRI2 shows no significant similarity with the only other P. ruminicola plasmid to have been studied previously, pRAM4.

Animals↗
PubMed2001

The effect of community-acquired pneumonia on plasma esterases in older people.

INTRODUCTION: Pneumonia is a major cause of morbidity and mortality in older people. The poor outcome of older pneumonia patients despite treatment is still not understood. OBJECTIVE: The aim of this study was to examine the effect of community-acquired pneumonia on enzymes of drug metabolism in older people. METHODS: Fifteen patients (median age 67 years) with a clinical and radiological diagnosis of community-acquired pneumonia and 14 healthy volunteers matched for age and gender (median age 75 years) were recruited. Plasma activities of benzoylcholinesterase, butyrylcholinesterase, acetylcholinesterase and aspirin esterase were determined spectrophotometrically at three time points in pneumonia patients--within 24 h of admission to hospital, 2 days later and 10 days later. Monocyte aryl hydrocarbon hydroxylase (AHH) activity was determined spectrofluorimetrically at the same time points. Enzyme activities were measured at one time point in healthy controls. RESULTS: Mean plasma benzoylcholinesterase activity was significantly lower in pneumonia patients on admission to hospital (mean +/- SEM 848 +/- 100) and after 10 days of treatment (mean +/- SEM 925 +/- 114) than in healthy controls (mean +/- SEM 1333 +/- 84, P < 0.05). Similarly, plasma acetylcholinesterase activity was significantly lower in pneumonia patients on admission (P = 0.007) and after 10 days of treatment (P = 0.01) than in controls. Butyrylcholinesterase activity was lower in pneumonia patients on admission (P = 0.029) than in healthy controls, but improved slightly after treatment so there was no longer a significant difference at 10 days compared with controls (P = 0.077). In contrast there were no significant differences in plasma aspirin esterase activity or induced monocyte AHH activity between pneumonia patients and healthy controls. The activities of benzoylcholinesterase (r = -0.536, P = 0.04), butyrylcholinesterase (r = -0.638, P = 0.01), acetylcholinesterase (r = -0.583, P = 0.022) and aspirin esterase (r = -0.624, P = 0.013) correlated inversely with the British Thoracic Society pneumonia poor prognostic index. CONCLUSION: The activities of several esterases are reduced in older pneumonia patients. Other enzymes including aspirin esterase and induced monocyte AHH activity are unaltered in pneumonia. There was a significant inverse relationship between the activities of all esterases studied and the British Thoracic Society pneumonia poor prognostic index.

Acetylcholinesterase↗
PubMed2001

Characterisation of novel point mutations in the survival motor neuron gene SMN, in three patients with SMA.

We report two novel mutations in three cases of spinal muscular atrophy (SMA), including two distant cousins who followed an unexpectedly severe course. Diagnosis was confirmed by reduced SMN protein and full-length SMN mRNA levels. Sequencing of the non-deleted SMN1 gene revealed a single G insertion at the end of exon 1 in the two cousins and a novel G275S exon 6 missense mutation in the milder case.

Child↗
PubMed2001

Woman-centred care.

Changes over the past decade have emphasised the individual service user and their relationship with the health service. Within the maternity services this has been interpreted as woman-centred care a result of key initiatives; the Winterton Report (House of Commons, 1992. Maternity Services. Second Report of the Health Committee (Winterton Report), Vol. 1. HMSO, London) and Changing Childbirth (DoH, 1993a, Changing childbirth: reports of the expert maternity group parts 1 & 2. HMSO, London). Changing Childbirth outlined key principles of the maternity services and the need for the woman (and her partner, if she wishes) to be the focus of care. The key principles are choice, continuity and control. High quality care depends on the recognition of individuals as having unique needs which continues to be reflected within contemporary policy documents (DoH, 1997, The new NHS: modern and dependable. HMSO, London). This paper presents findings related to the provision of woman-centred care from a national research and development study. The study design incorporated (i): a national survey which was undertaken with midwives, midwife supervisors and doctors; and (ii): in-depth case studies in which information was obtained through interviews with midwives, midwife supervisors, educators, managers, doctors and mothers. Midwives, at all levels, are involved in changing maternity service provision and adapting to new systems of care which aim to increase continuity of care and carer for the woman. The researchers sought to understand how woman-centred care was interpreted and experienced in practice. The findings have been used to identify the continuing educational needs of midwives, and to develop an open learning educational package to meet identified need. The curriculum was designed to enhance the move towards the provision of a more integrated woman-centred service.

Adult↗
PubMed2001

Craniofacial approaches to tumors of the anterior skull base.

An approach that combines extracranial and intracranial components-termed a craniofacial approach-allows en bloc extirpation of paranasal malignancies that abut or penetrate the skull base. When combined with radiotherapy, cure rates for such tumors rose from near zero in the 1950s to 39% to 86% by 2000, with the higher rates reflective of esthesioneuroblastomas, well-differentiated adenocarcinomas, vasoformative tumors, and meningiomas. Transfacial tumor access can involve a transnasal (endoscopic or with magnification loupes) exposure, a midfacial degloving, or a lateral rhinotomy, depending on tumor location and size. Adjunctive exposures for tumors penetrating the nasopharynx, pterygomaxillary fossa, or sphenoid include the lateral facial split and the mandibular swing. The standard transcranial accesses are a sub-basal variation of frontal craniotomy, which encompasses en bloc the superior orbital rims, the nasion, and the lower frontal bones; or, a frontotemporal craniotomy with mobilization of the lateral orbital rim and the zygoma.

Humans↗
PubMed2001

Jugulotympanic paragangliomas.

Jugulotympanic paragangliomas are the most common tumors of the middle ear and temporal bone. Although these larger tumors can prove to be formidable, the advent of microscopic and skull base surgical techniques has greatly enhanced the ability to treat and manage these tumors.

Glomus Jugulare Tumor↗
PubMed2001

The role of central and peripheral Cannabinoid1 receptors in the antihyperalgesic activity of cannabinoids in a model of neuropathic pain.

We have examined the effects of cannabinoid agonists on hyperalgesia in a model of neuropathic pain in the rat and investigated the possible sites of action. The antihyperalgesic activity of the cannabinoids was compared with their ability to elicit behavioural effects characteristic of central cannabinoid activity. WIN55,212-2 (0.3-10 mg kg(-1)), CP-55,940 (0.03-1 mg kg(-1)) and HU-210 (0.001-0.03 mg kg(-1)) were all active in a 'tetrad' of tests consisting of tail-flick, catalepsy, rotarod and hypothermia following subcutaneous administration, with a rank order of potency in each of HU-210 > CP-55,940 > WIN55,212-2. The effects of WIN55,212-2 in each assay were blocked by the Cannabinoid1 (CB1) antagonist SR141716A. In the partial sciatic ligation model of neuropathic pain WIN55,212-2, CP-55,940 and HU-210 produced complete reversal of mechanical hyperalgesia within 3 h of subcutaneous administration with D50 values of 0.52, 0.08 and 0.005 mg kg(-1), respectively. In this model WIN55,212-2 was also effective against thermal hyperalgesia and mechanical allodynia. WIN55,212-2 produced pronounced reversal of mechanical hyperalgesia following intrathecal administration that was blocked by the CB1 antagonist SR141716A. Following intraplantar administration into the ipsilateral hindpaw, WIN55,212-2 produced up to 70% reversal of mechanical hyperalgesia, although activity was also observed at high doses following injection into the contralateral paw. The antihyperalgesic effect of WIN55,212-2 injected into the ipsilateral paw was blocked by subcutaneously administered SR141716A, but was not affected by intrathecally administered SR141716A. These data show that cannabinoids are highly potent and efficacious antihyperalgesic agents in a model of neuropathic pain. This activity is likely to be mediated via an action in both the CNS and in the periphery.

Analgesics↗
PubMed2001

Coordination of Ca2+ signalling by NAADP.

Nicotinic acid adenine dinucleotide phosphate (NAADP) mobilizes intracellular Ca2+ stores in several cell types. Ample evidence suggests that NAADP activates intracellular Ca2+ channels distinct from those that are sensitive to inositol trisphosphate and ryanodine/cyclic ADP-ribose. Recent studies in intact cells have demonstrated functional coupling ('channel chatter') between Ca2+ release pathways mediated by NAADP, inositol trisphosphate and cyclic ADP-ribose. Thus, NAADP is probably an important determinant in shaping cytosolic Ca2+ signals.

Calcium Signaling↗
PubMed2001

An in vivo binding assay to determine central alpha(1)-adrenoceptor occupancy using [(3)H]prazosin.

An alpha(1) adrenoceptor (alpha(1)-AdR) assay using [(3)H]prazosin binding in mouse brain is described which allows in vivo determination of central alpha(1)-AdR occupancy for ligands with alpha(1)-AdR affinity. Binding of [3H]prazosin in rat and mouse brain membranes in vitro was used to characterise the pharmacological profile of alpha(1)-AdRs in order to determine any potential species variations. Saturation and displacement studies yielded comparable affinity and pharmacological profile for [(3)H]prazosin binding in mouse and rat brain homogenates. These studies confirmed the absence of species variation for ligands in central alpha(1)-AdR pharmacology which is in good agreement with previous studies in rat brain. Subsequently, in vivo binding of [(3)H]prazosin in mouse whole brain was used to measure the occupancy of a number of AdR ligands. Timecourse studies revealed that a [3H]prazosin (5 mu Ci/mouse) pretreatment time of at least 20 min following intravenous (i.v.) administration was required for optimal specific binding. Ligands were administered systemically 40 min prior to i.v. administration of radiolabel. The alpha(1)-adrenoceptor ligands prazosin (ED(50)=0.15 mg/kg i.p.), benoxathian (0.52 mg/kg i.p.) and phentolamine (51 mg/kg i.p.) were all able to block in vivo [(3)H]prazosin binding from mouse brain. In addition, receptor occupancy values for a number of compounds including haloperidol (ED(50)=0.83 mg/kg s.c.), clozapine (2.2 mg/kg s.c.) and MDL-100907 [R(+)-alpha-(2,3-dimethoxyphenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidine-methanol], (10 mg/kg s.c.)], which possess high to moderate affinity at alpha(1)-adrenoceptors, were also determined. These results suggest that in the mouse, [(3)H]prazosin binding can be used to measure in vivo receptor occupancy of ligands with affinity at central alpha(1)-adrenoceptors.

Adrenergic alpha-Antagonists↗