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Biomedical subjects

D R Patel

Publications and source records attributed to D R Patel.

At least 37 records · Page 2Linked to original sources

Evidence for a role of presynaptic AMPA receptors in the control of neuronal glutamate release in the rat forebrain.

The role of presynaptic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in controlling the neuronal release of excitatory amino acids has been investigated. Stimulation of presynaptic AMPA receptors by the endogenous agonist L-glutamate, or by (R,S)-AMPA, dose-dependently enhanced the Ca(2+)-dependent, tetrodotoxin-insensitive, electrically-stimulated release of [3H]D-aspartate from rat forebrain slices. This AMPA receptor-mediated response showed marked stereoselectivity with the activity residing solely in the (S)-isomer. (R)-AMPA was inactive in this respect. AMPA-evoked responses were significantly enhanced in the presence of the AMPA receptor desensitization inhibitor, cyclothiazide (10 microM). Moreover, responses to both AMPA and glutamate were inhibited by competitive (NBQX) and non-competitive (GYKI 52466) AMPA receptor-selective antagonists in a dose-dependent manner. These results provide strong support for the existence of presynaptic AMPA receptors acting to enhance the synaptic release of excitatory amino acids in the mammalian forebrain. Such a positive feedback system may play an important functional role in physiological (e.g., long-term potentiation) and/or pathological (e.g., epileptogenesis) processes in the mammalian central nervous system. AMPA-type autoreceptors may provide new targets for drug action.

Animals↗

The rebellious adolescent. Evaluation and management of oppositional and conduct disorders.

A wide variety of management options are available to the primary care physician who is presented with a rebellious adolescent. After a careful assessment, the clinician and other health care professionals can choose a diverse combination of interventions: individual therapy, family therapy, youth-centered programs, community-centered programs, psychopharmacology, and others. Rebellious adolescents need access to comprehensive medical and mental health care, academic education (including sexuality education), and full employment opportunities. Primary care physicians can play a vital and sometimes pivotal role coordinating services and helping parents, school personnel, and therapists work with these youth. Even when dealing with the very difficult and resistant group of youth with CD and ODD, optimism for improvement should always be maintained by the clinician.

Adolescent↗

Molecular cloning and expression of a human secretin receptor.

Secretin is a 27-amino acid neuroendocrine peptide that stimulates fluid and electrolyte secretion in the gastrointestinal tract, activates tyrosine hydroxylase activity in the central nervous system, and affects cardiac and renal function. Specific receptors for secretin have been previously characterized on neuroblastoma cells, pancreatic acini, gastric glands, and liver cholangiocytes. We report here the isolation of a 1616-base pair cDNA from human lung tissue that encodes a 440-amino acid, 50-kDa, G protein-coupled human secretin receptor (HSR), with homology of 80% with the rat secretin receptor and 37% with the human type I vasoactive intestinal peptide receptor. Northern blot analysis of human tissue mRNA revealed that the relative intensity for expression of a 2.1-kilobase HSR transcript was pancreas > kidney > small intestine > lung > liver, with trace levels in brain, heart, and ovary. Stable transfectants of HSR in human embryonic kidney 293 cells, termed 293S12, expressed 10(5) binding sites/cell for 125I-secretin, with an apparent Kd of 3.2 nM. Vasoactive intestinal peptide, pituitary adenylyl cyclase-activating peptide-38, and glucagon were less potent (by 3 orders of magnitude) than secretin in competitively inhibiting 125I-secretin binding to 293S12 cells. Secretin evoked concurrent dose-dependent increases in intracellular cAMP and calcium levels in 293S12 cells and stimulated a 4-fold increase in phosphatidylinositol hydrolysis. Thus, the HSR expressed by stable transfectants can couple to two distinct intracellular signaling pathways.

Amino Acid Sequence↗

Human vasoactive intestinal peptide1 receptors expressed by stable transfectants couple to two distinct signaling pathways.

Vasoactive intestinal peptide (VIP) is a potent neuropeptide mediator of central and peripheral nervous system function. A human VIP1 receptor (HVR) cDNA clone was previously obtained from HT29 intestinal epithelial cells and lung tissue. Stably-transfected human embryonic kidney 293 cells and chinese hamster ovary (CHO) cells expressing about 10(6) HVRs per cell that bind [125I]VIP with a Kd of 0.2-0.8 nM, and specifically recognized by anti-HVR antibodies, were established and characterized. VIP induced increases in intracellular cAMP levels ([cAMP]i) dose-dependently with an EC50 of 0.2 nM in 293 and CHO stable transfectants and concurrently evoked dose-dependent increases in intracellular calcium concentrations ([Ca2+]i), as determined by fluorescence-dye spectroscopy. Untransfected 293 and CHO cells showed minimal binding or intracellular effects of VIP; however, native VIP1 receptors of HT29 cells also increased [cAMP]i and [Ca2+]i in dose-dependent responses to VIP. Thus recombinant and native human VIP1 receptors both couple to two distinct signal transduction pathways within a single cell type.

Animals↗

Specific recognition of the human neuroendocrine receptor for vasoactive intestinal peptide by anti-peptide antibodies.

Rabbit polyclonal IgG antibodies were generated to three distinct synthetic peptide substituents of the human neuroendocrine-type 7 transmembrane-domain receptor for vasoactive intestinal peptide (VIP), including a portion of the amino-terminus, first extracellular loop, and carboxyl-terminus. Immunofluorescent staining of both human K293 cell transfectants, expressing recombinant VIP receptors, and HT-29 human intestinal epithelial cells, bearing native VIP receptors, was observed with each of the antibodies and was eliminated specifically after absorption of antibodies with the respective peptide immunogen. Each of the antibodies recognized the same approximately 70-kDa membrane proteins, extracted from both K293 cell transfectants and HT-29 cells, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis blots. Neither IgG nor Fab preparations of the antibodies inhibited VIP binding to cellular receptors at a concentration of 1 microgram/ml, that yielded optimal immunofluorescence, or at 5-300 micrograms/ml. In contrast, 5-200 micrograms/ml of anti-peptide antibodies as IgG, but not Fab, significantly inhibited the increase in concentration of cyclic AMP in HT-29 cells elicited by 1 nM VIP, without affecting the greater increase evoked by 100 nM VIP or alone altering the level of cyclic AMP. Antibodies to several peptide substituents thus bind specifically to VIP receptors in immunoblots and permeabilized cells, and may affect the cellular functions of VIP receptors with sufficient selectivity to reduce transduction of signals, without altering the binding of VIP.

Amino Acid Sequence↗

Stem cell factor is a neurotrophic factor for neural crest-derived chick sensory neurons.

We have found that stem cell factor (SCF) selectively enhances the survival of cultured embryonic chick dorsal root ganglia (DRG) neurons. Neurons grown in the presence of SCF expressed both neurofilament 150 kDa subunit and calcitonin-gene related peptide. SCF does not, however, enhance the survival of parasympathetic, placode-derived sensory or sympathetic neurons in culture. Combining SCF with brain-derived neurotrophic factor or neurotrophin-3, but not with NGF, maintains more neurons than either factor alone, suggesting that these factors have partially overlapping activities. SCF preferentially rescues small neurons from the DRG. Labeling studies with bromodeoxyuridine indicate that the neurons sustained by SCF are not differentiating from a dividing progenitor.

Animals↗

Cloning and functional expression of a human neuroendocrine vasoactive intestinal peptide receptor.

Vasoactive intestinal peptide (VIP) is a potent mediator of gastrointestinal, nervous, pulmonary, vascular and immune functions. A cDNA was obtained from human HT29 intestinal epithelial cells and found to encode a 457 amino acid, 52 kDa VIP receptor. Transfection of the cDNA into COS-7 cells and 293 cells resulted in expression of specific saturable binding of VIP with a Kd of 0.8 nM, and induction of increases in intracellular cAMP by VIP with an EC50 of 1 nM. The human VIP receptor is homologous to other G protein-coupled receptors of the secretin-parathyroid hormone receptor family. A 2.8 kb transcript was detected in human lung, HT29 cells and Raji B-lymphoblasts with weaker expression in human brain, heart, kidney, liver and placenta.

Amino Acid Sequence↗

The role of oriT in tra-dependent enhanced recombination between mini-F-lac-oriT and lambda plac5.

Recombination between F42lac and lambda plac5 is typically 20- to 50-fold more efficient than recombination between chromosomal lac and lambda plac5. This enhancement of recombination is recBCD-dependent and requires the expression of genes from the tra regulon of the F factor. Also required is oriT, the origin of F factor conjugational transfer, which must be located in-cis to the cellular copy of lac. In this study we show that enhanced recombination is not supported by an oriT point mutant that reduces oriT function in conjugation. We also present evidence that the activation of oriT for recombination enhancement involves the same strand-specific nick that is required for conjugal DNA transfer. Although it is thought that the role of oriT in recombination enhancement is related to the facilitated entry of RecBCD enzyme into the DNA duplex, we were unable to detect any double-strand breakage at oriT.

Base Sequence↗

Primary prostatic involvement in non-Hodgkin lymphoma.

We report 3 cases of primary extranodal lymphoma of the prostate, an unusual extranodal presentation rarely diagnosed antemortem. Symptoms of prostatism associated with an enlarged hard prostate with pyuria and hematuria in younger patients should suggest the diagnosis. Urine cytologic examination should aid in the diagnosis of this condition.

Adult↗

Thermodynamic proton-ligand and metal-ligand stability constants of some drugs.

The thermodynamic proton-ligand (pKa) and metal-ligand stability constants of clioquinol, clofibrate, nitrofurazone, and tetracycline with Cu2+, Zn2+, Mn2+, Mg2+, and Ca2+ have been determined at 35 degrees C in 50% ethanol-water media. An empirical pH correction for mixed-aqueous media has been applied. The metal-ligand stability constants were determined by following the Bjerrum Calvin titration technique as applied by Agrawal to mixed-aqueous solvents. The effect of the basicity of the ligand and the order of stability constants is discussed. The stability constants of the divalent metals follow the order: Cu2+ greater than Zn2+ greater than Mn2+ greater than Mg2+ greater than Ca2+ with all the drugs.

Buffers↗

T cell subsets in idiopathic glomerulonephritis.

Determination of T lymphocyte subsets using monoclonal antibodies revealed a deficiency of suppressor cells and an elevated helper:suppressor T cell ratio in 7 patients with untreated idiopathic immune complex glomerulonephritis and nephrotic syndrome. Possible pathogenetic mechanisms of immunoregulatory cell imbalance and their implications specifically in reference to idiopathic glomerulonephritis are discussed.

Adult↗