Familial testotoxicosis.
Explore the source record for details and available documents.
Child, Preschool↗
Biomedical subjects
P Banerjee
Publications and source records attributed to P Banerjee.
Emerging facets of diastolic heart failure.
One hundred patients with congestive cardiac failure (52 males and 48 females) with age ranging from 16 to 56 yrs (mean age 42 +/- 6) were studied to determine the relative prevalence of systolic and diastolic failures, their clinical profiles and common aetiologies. Age matched 25 control subjects were also studied to established a normal range of echocardiographic values for LV diastolic function. Thirty eight patients (38%) were found to have pure diastolic heart failure and another 5 (5%) and 57 (57%) were detected to have mixed and systolic failures respectively. An attempt to correlate the clinical assessment of diastolic failure with echo doppler study showed the sensitivity and specificity of the clinical criteria for diagnosis of diastolic heart failure to be 100% and 91.94% respectively. Of the 38 cases of diastolic failure detected 39.5% had hypertension, 31.6% ischaemic heart disease and 13.16% hypertrophic cardiomyopathy.
Adolescent↗
Clinical evaluation of a herbal antidiabetic product.
Sixty-seven diabetic patients and 12 normal subjects were selected for a clinical study with an indigenous herbal product. The study consisted of 2 phases. In phase 1 study out of 25 diabetics (both insulin dependent and non-insulin dependent) only those in the age group of 41-50 years ie, 11 cases showed lowering of mean high blood sugar level in all samples from 1/2 an-hour to 2 hours with the test drug containing guar gum, methi, tundika and mesha shringi. But in phase 2 study there was lowering of blood sugar level with the test drug and with 2 of its constituents ie, guar gum and methi when used separately in 42 non-insulin dependent diabetics. While there was some blood sugar level lowering effect with guar gum and methi when used separately in 12 normal subjects in phase 2 study, but that was not the same observed with the test drug. The results of this study indicate the efficacy of the product as an adjuvant.
Adult↗
Differential solubilization of membrane lipids by detergents: coenrichment of the sheep brain serotonin 5-HT1A receptor with phospholipids containing predominantly saturated fatty acids.
Most membrane receptors lose binding activity during purification and we studied the correlation between this event and differential solubilization of membrane lipids by detergents. Both 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate [Chaps; high critical micellar concentration (cmc) approximately 0.5%] and n-dodecyl beta-D-maltoside (DDM; low cmc approximately 0.009%) solubilized the 8-[3H]hydroxy-2-(di-n-propylamino)tetralin ([3H]8-OH-DPAT)-binding serotonin 5-HT1A receptor (5-HT1A-R) optimally at 2% (w/v) detergent concentration despite the widely differing cmc of the two detergents. In contrast, n-octyl-beta-D-glucopyranoside (octyl glucoside; high cmc approximately 0.7%), Thesit (low cmc approximately 0.005%), and Triton X-100 (low cmc approximately 0.013%) solubilized virtually no [3H]8-OH-DPAT-binding activity. The total mass of solubilized lipids was always low at 0.5% detergent concentration and attained a plateau between 1 and 2.5% for all detergents except octyl glucoside. The mass of octyl glucoside-solubilized lipids showed an increasing trend even at 3.0% detergent concentration. The total amount of solubilized lipid is unrelated to the amount of 5-HT1A-R solubilized but the species of lipid is important. Thus Chaps and DDM, with diverse structures and cmc, both preferentially solubilized phospholipids enriched in saturated fatty acids (67 and 72%, respectively). In contrast, octyl glucoside, Triton X-100, and Thesit showed no preference in solubilizing phospholipids. Octyl glucoside, which solubilized significantly higher proportions of saturated fatty-acid-containing phosphatidylethanolamine (slightly higher saturated fatty acids in total phospholipids), also produced more (twofold higher) solubilized 5-HT1A sites than Triton X-100 and Thesit. This suggests an optimum involvement of saturated fatty acid side chains in forming tightly packed vesicles which stabilize the 5-HT1A-R more than the vesicles of higher fluidity formed by phospholipids containing higher proportions of cis-double-bonded unsaturated fatty acids. Indeed, delipidation of the 1.5% Chaps-solubilized receptor preparations by Sephacryl S-200 column chromatography eliminated essentially all [3H]8-OH-DPAT-binding activity. Therefore, for efficient recovery during solubilization and reconstitution of a prototypic heptahelical receptor (5-HT1A), it is essential to stabilize the receptor protein through association with saturated phospholipids.
8-Hydroxy-2-(di-n-propylamino)tetralin↗
Regulation of an epitope-tagged recombinant Rsk-1 S6 kinase by phorbol ester and erk/MAP kinase.
Phorbol ester tumor promoters (TPA) activate the endogenous erk/MAP kinases and Rsk S6 kinases but not the p70S6 kinase in COS cells. DNA sequences encoding the rat Rsk-1 S6 kinase (homologous to Xenopus rsk alpha), modified by insertion of a peptide epitope at the polypeptide aminoterminus, were expressed transiently in COS cells. TPA stimulates the 40S and peptide kinase activity of the recombinant epitope-tagged Rsk-1, as well as the extent of Rsk-1 autophosphorylation in vitro (32P-Ser >> 32P-Thr). Indications that the conformation of the recombinant Rsk-1 polypeptide is substantially changed after activation by TPA in situ include a retarded mobility of the Rsk-1 polypeptide on SDS-PAGE and the appearance of new 32P-peptides during autophosphorylation in vitro. All these features of the TPA-activated Rsk-1 S6 kinase are abolished by dephosphorylation of the kinase in vitro with Ser/Thr phosphatase-2A. TPA increases 32P incorporation into recombinant Rsk-1 by 2-3-fold (32P-Ser >> 32P-Thr). Peptide mapping exhibits a single major 32P-peptide in Rsk-1 isolated from unstimulated cells and 10-12 additional 32P peptides after TPA treatment in situ. Phosphorylation of basal or phosphatase-2A-treated recombinant Rsk-1 in vitro with erk2/MAP kinase increases Rsk-1 40S kinase, peptide kinase, and autophosphorylating activity, retards migration of Rsk-1 polypeptides on SDS-PAGE, and generates new sites of Rsk-1 autophosphorylation in vitro. By contrast, TPA-activated Rsk-1 is not altered in these properties by autophosphorylation in vitro. By contrast, TPA-activated Rsk-1 is not altered in these properties by phosphorylation in vitro with erk2/MAP kinase. Activation of Rsk-1 in situ with TPA diminishes by over 90% the extent of Rsk-1 phosphorylation achieved in vitro by erk2/MAP kinase, as compared to the parallel phosphorylation of a phosphatase-2A-treated Rsk-1; basal Rsk-1 is intermediate. Peptide maps of phosphatase-2A-treated Rsk-1 after phosphorylation in vitro with erk2/MAP kinase exhibit 32P-peptides that comigrate with nearly all of the 32P-peptides present in TPA-activated-32P Rsk-1 labeled in situ, plus several 32P-peptides characteristic of Rsk-1 autophosphorylation in vitro.(ABSTRACT TRUNCATED AT 400 WORDS)
Amino Acid Sequence↗
Heterologous expression of the serotonin 5-HT1A receptor in neural and non-neural cell lines.
Stable expression of neuronal receptors in cell lines of neural origin is important for studies of neurotransmitter mediated signal transduction. We have achieved this for the first time in three cell lines which are derived from various tissues of neural origin (hippocampus, HN2; chinese hamster brain explant, NCB-20; rat dorsal root ganglion, F-11). Following electroporation assisted transfer of a construct containing the hippocampal serotonin 5-HT1A receptor (5-HT1AR) DNA, one neural cell line, NG-108-15 (murine neuroblastoma x C6 glioma), failed to express the transfected activity, while three others as well as the non-neural CHO (chinese hamster ovary) cells expressed high levels of the receptor. Upon normalization to coexpressed human beta-hexosaminidase B activity, it was found that the human 5-HT1AR, which is normally concentrated in the hippocampus and at a lesser density in the brain, was expressed at the highest level (15.7 x 10(4) receptors/cell) in the HN2 followed by the NCB-20 (8.3 x 10(4) receptors/cell), F-11 (4.4 x 10(4) receptors/cell) and lastly the non-neuronal CHO (4.2 x 10(4) receptors/cell) cells. Ten-twelve days after passage, a striking increase in expression of the receptor was observed only in the cell lines of neural origin. By contrast, there was no appreciable increase in expression of the transfected 5-HT1AR in the non-neural CHO cells over time. This late increase in expression was eliminated in cells which had been maintained in low glucose (1 g/L) for the first two days after passage, thus establishing a vital role of glucose in expression of the transfected 5-HT1AR in cell lines of neural origin. In all cases the 5-HT1AR was negatively coupled to adenylate cyclase, as evidenced by an agonist mediated decrease in prostaglandin E1 stimulated cyclic AMP levels.
8-Hydroxy-2-(di-n-propylamino)tetralin↗
Detection of structural and trace metal changes in scalp hair of radiographers.
Human scalp hair samples were collected from 24 medical radiographers and from 24 nonradiographers (control group) of matching age groups belonging to both sexes. Changes in zinc, copper, and iron content, as well as structural morphology of hair, were studied by atomic absorption spectrometry and scanning electron microscopy, respectively. Such studies revealed increases in zinc, copper, and iron concentrations along with varied types of cuticular damage in the radiographers' hair compared to that of nonradiographers. Present observations indicate the possibility of radiation hazards for radiation workers.
Adult↗
Role of lipids in receptor mediated signal transduction.
A new strategy has been successfully applied to reconstitute the brain specific serotonin 5-HT1A receptor-G protein-adenylate cyclase complex. A mild method of tissue preparation gave a stable, membrane-bound form of the receptor (SBP) which retained its natural lipid content. Treatment of SBP with serotonin (1 microM) and 3-[(3-cholamidopropyl) dimethyl ammonio]-1-propanesulphonate (CHAPS) (2%) solubilized the ligand-receptor-G protein-ligand complex along with the associated phospholipids and cholesterol. Dialysis of this extract (SBDS) against buffer containing 25% ethylene glycol produced a stable, reconstituted and active preparation (SBDSE) of vesicles which upon centrifugal separation followed by gentle resuspension retained 95-100% [3H] 8-OH-DPAT binding activity as well as 60% [3H] GppNHp binding and adenylate cyclase activities of SBDSE. The reconstituted receptor preparation compared well with the membrane-bound form in displaying a similar value for KD (2.1 nM) and a single affinity state for [3H] 8-OH-DPAT binding (Bmax = 118 fmol/mg). However, in sharp contrast to the membrane-bound receptor which was negatively coupled to adenylate cyclase, agonist treatment of the solubilized and reconstituted receptor resulted in an increase in adenylate cyclase. This change in receptor-adenylate cyclase coupling following reshuffling of membrane lipids during solubilization and reconstitution suggested that membrane lipids could have a profound effect on receptor-effector coupling. To study the effect of membrane lipid composition on receptor-mediated signal transduction in a stabler and more natural system, neural cells derived from different parts of the brain (hippocampus, HN2; CNS, NCB-20; dorsal root ganglion, F-11) and a non-neural cell line (CHO), all with differing membrane lipid compositions, were selected. Since no known cell line contains the serotonin 5-HT1A receptor (5-HT1A-R), stable transfection of the selected cell lines with a DNA construct encoding the human 5-HT1A-R was carried out and this resulted in a late increase of [3H] 8-OH-DPAT binding in the stationary phase only in the cell lines of neural origin. In the non-neural cell line (CHO), which also displayed marked difference in membrane lipids, the receptor was positively coupled to the phospholipase C-IP3-[Ca2+]i cascade. Even though GPLC was present in the NCB-20 and F-11 cells as evidenced by a bradykinin receptor-mediated increase in inositol phosphates in these cells 8-OH-DPAT treatment resulted in no change in phospholipase C in any of the cell lines of neural origin.(ABSTRACT TRUNCATED AT 400 WORDS)
8-Hydroxy-2-(di-n-propylamino)tetralin↗
Nephrotic syndrome in non-cirrhotic portal fibrosis: a report of three cases.
We describe three patients with nephrotic syndrome and non-cirrhotic portal fibrosis. In two of them, the liver disorder preceded the nephrotic syndrome while in the third patient the two conditions presented together. All three patients achieved remission of nephrotic syndrome with prednisolone and cyclophosphamide.
Adolescent↗
Enrichment of saturated fatty acid containing phospholipids in sheep brain serotonin receptor preparations: use of microwave irradiation for rapid transesterification of phospholipids.
During enrichment of the 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT)-binding serotonin 5-HT1A receptors from sheep brain gray matter (membrane isolation, detergent solubilization and reconstitution into vesicles) a consistent and striking increase in the composition of saturated fatty acids was observed in phospholipids which were coisolated with the receptors. A rapid procedure has been developed for the methylation of free and phospholipid linked fatty acids which were thus analyzed by gas chromatography-mass spectrometry (GC/MS). Esterification of free fatty acids and transesterification of phospholipid linked fatty acids were achieved with 14% boron trifluoride in methanol (BF3-CH3OH) in 20 s and 50 s, respectively, under low power microwave irradiation (60 W) with a post-reaction cooling of less than 5 min. This is in contrast to the conventional method of heating in a boiling water bath for 10-15 min with BF3-CH3OH which is inevitably preceded by time-consuming and inconvenient clamping of vials and followed by cooling for 10 min before the vials can be safely opened. Analysis of fatty acid profiles in phosphatidylethanolamine (PE) and phosphatidylcholine (PC) from egg yolk, phosphatidylinositol (PI) from bovine liver and phosphatidylserine (PS) from bovine brain by both techniques showed comparable results. During detergent solubilization of sheep brain gray matter, the overall proportion of saturated fatty acids in PE (major lipid), PI, PC (major lipid) and PS increased from 50-60% in sheep brain phospholipids to 70-75% in 1.5% CHAPS solubilized, reconstituted and biologically active serotonin 5-HT1A preparations. In sharp contrast, the proportions of saturated fatty acids in 1.5% Triton X-100 solubilized PE (48.1%) (major lipid), PI (63.6%), PC (60.6%) (major lipid) and PS (62.2%) were not significantly different from those in the original sheep brain membranes. Strikingly, this was coupled with the occurrence of very low levels of 5-HT1A receptor activity in the Triton X-100 solubilized preparations. The abundance of 5-HT1A sites in the enriched vesicles obtained only from the CHAPS-solubilized preparations was further confirmed by specific radiolabeling of a 58-kDa polypeptide by the 5-HT1A specific ligand p-aminophenylethyl-m-trifluoromethylphenylpiparazine (PAPP) which was coupled to a 125I-labeled, photoreactive, heterobifunctional cross-linker, sulfosuccinimidyl-2-(p-azidosalicylamido)ethyl-1,3'-dithiopropiona te (SASD). Thus CHAPS-solubilized 5-HT1A receptor preparations are depleted in the more rigid lipids such as sphingolipids and cholesterol, (Banerjee et al. (1990) Biochim. Biophys. Acta 1044, 305-314), but are enriched in vesicle-stabilizing, phospholipid-linked saturated fatty acids which in turn probably stabilize the heptahelical, membrane bound 5-HT1A receptor.
8-Hydroxy-2-(di-n-propylamino)tetralin↗
Raf-1 activates MAP kinase-kinase.
The normal cellular homologue of the acutely transforming oncogene v-raf is c-raf-1, which encodes a serine/threonine protein kinase that is activated by many extracellular stimuli. The physiological substrates of the protein c-Raf-1 are unknown. The mitogen-activated protein (MAP) kinases Erk1 and 2 are also activated by mitogens through phosphorylation of Erk tyrosine and threonine residues catalysed by a protein kinase of relative molecular mass 50,000, MAP kinase-kinase (MAPK-K). Here we report that MAPK-K as well as Erk1 and 2 are constitutively active in v-raf-transformed cells. MAPK-K partially purified from v-raf-transformed cells or from mitogen-treated cells can be deactivated by phosphatase 2A. c-Raf-1 purified after mitogen stimulation can reactivate the phosphatase 2A-inactivated MAPK-K over 30-fold in vitro. c-Raf-1 reactivation of MAPK-K coincides with the selective phosphorylation at serine/threonine residues of a polypeptide with M(r) 50,000 which coelutes precisely on cation-exchange chromatography with the MAPK-K activatable by c-Raf-1. These results indicate that c-Raf-1 is an immediate upstream activator of MAPK-K in vivo. To our knowledge, MAPK-K is the first physiological substrate of the c-raf-1 protooncogene product to be identified.
3T3 Cells↗
Evidence for lipase abnormality: high levels of free and triacylglycerol forms of unsaturated fatty acids in neuronal ceroid-lipofuscinosis tissue.
Total lipid obtained from normal and different forms of neuronal ceroid-lipofuscinoses (NCL) tissues was analyzed by high performance thin layer chromatography (HPTLC). We observed a large (greater than 6-fold) increase in a lipid band corresponding to triolein for NCL dog pancreas and spleen and juvenile human NCL brain and infantile NCL spleen. The accumulation was less pronounced for the brain samples but apart from increased dolichol-monophosphate levels, other lipids appeared normal. Normal dog, goat, or human spleen contained virtually no triacylglycerol, and of the pathological controls, beta-mannosidosis goat spleen showed no triacylglycerol band at all. A sample of human spleen from a patient with lymphoma-associated splenomegaly displayed a strong triacylglycerol band, but gas chromatography-mass spectrometry (GC/MS) of the bands showed an equal increase in both saturated and unsaturated fatty acid containing triacylglycerols in the splenomegaly sample, in keeping with the notion of non-specific fat deposition in damaged tissue. In contrast, in all the NCL samples (spleen, pancreas, and brain) a prominent increase in the proportion of unsaturated fatty acids was observed in both free fatty acid and/or triacylglycerol bands following GC/MS. The NCL-English setter dog pancreas showed a major presence of oleic acid (18:1) (twofold increase) as compared to normal, while dog and infantile human NCL spleen samples and juvenile Batten brain (human) displayed a robust increase in linoleic acid (18:2) and sometimes in oleic acid and arachidonic acid (20:4) (for infantile human NCL spleen). For the infantile human NCL spleen sample an increase in linoleic acid in both free fatty acid (3.2-fold) and triacylglycerol (10-fold) was observed.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Molecular basis of an adult form of beta-hexosaminidase B deficiency with motor neuron disease.
A patient (KL) with progressive motor neuron disease associated with partial Hex A (alpha beta) and no Hex B (beta beta) activity, synthesized beta-chains which only associated with alpha-chains. To identify the molecular basis of this inability of beta-chains to self associate, RNA from cultured fibroblasts was reverse transcribed, the cDNA encoding the beta-chain amplified by polymerase chain reaction, subcloned, and sequenced to reveal two types of single missense mutation. The first mutation, (Type I) 619A----G, was paternally inherited and converted a 207IIe----Val in a highly conserved region believed to be associated with catalytic activity and activator protein binding. Biochemical evidence for impaired activator protein binding was obtained by purifying Hex A from KL urine and demonstrating a greater than 50% reduction of in vitro GM2 hydrolysis compared to normal urinary Hex A. In other cDNA species (Type II), a maternally inherited 1367A----C mutation converted 456Tyr----Ser in another highly conserved region of the beta-chain and we propose that this mutation leads to the inability of the beta-chains to self associate and thus reach maturity. These same cDNA species contained a second 362A----G mutation which converted 121Lys----Arg, but is apparently a polymorphism since it also occurs in some normal subjects. We propose that the patient is a compound heterozygote in which a combination of no self-association of the mutant beta-chains and impaired activator protein binding to alpha-beta (mutant) (Hex A) required for GM2 hydrolysis result in total beta-Hex B deficiency and slow accumulation of GM2 ganglioside, primarily in motor neurons.
Adult↗
Properties of high spin states in 53Cr and 53Mn.
Explore the source record for details and available documents.
Journal Article↗
Projectile breakup by nuclear and Coulomb fields and application to astrophysically relevant radiative-capture processes.
Explore the source record for details and available documents.
Journal Article↗
Cloning and expression of two human p70 S6 kinase polypeptides differing only at their amino termini.
Two classes of human cDNA encoding the insulin/mitogen-activated p70 S6 kinase have been isolated; the two classes differ only in the 5' region, such that the longer polypeptide (p70 S6 kinase alpha I; calculated Mr 58,946) consists of 525 amino acids, of which the last 502 residues are identical in sequence to the entire polypeptides encoded by the second cDNA (p70 S6 kinase alpha II; calculated Mr 56,153). Both p70 S6 kinase polypeptides predicted by these cDNAs are present in p70 S6 kinase purified from rat liver, and each is thus expressed in vivo. Moreover, both polypeptides are expressed from a single mRNA transcribed from the (longer) p70 S6 kinase alpha I cDNA through the utilization of different translational start sites. Although the two p70 S6 kinase polypeptides differ by only 23 amino acid residues, the slightly longer alpha I polypeptide exhibits anomalously slow mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), migrating at an apparent Mr of 90,000 probably because of the presence of six consecutive Arg residues immediately following the initiator methionine. Transient expression of p70 alpha I and alpha II S6 kinase cDNA in COS cells results in a 2.5- to 4-fold increase in overall S6 kinase activity. Upon immunoblotting, the recombinant p70 polypeptides appear as a closely spaced ladder of four to five bands between 65 and 70 kDa (alpha II) and 85 and 90 kDa (alpha I). Transfection with the alpha II cDNA yields only the smaller set of bands, while transfection with the alpha I cDNA generates both sets of bands. Mutation of Met-24 in the alpha I cDNA to Leu or Thr suppresses synthesis of the alpha II polypeptides. Only the p70 alpha I and alpha II polypeptides of slowest mobility on SDS-PAGE comigrate with the 70- and 90-kDa proteins observed in purified rat liver S6 kinase. Moreover, it is the recombinant p70 polypeptides of slowest mobility that coelute with S6 kinase activity on anion-exchange chromatography. The slower mobility and higher enzymatic activity of these p70 proteins is due to Ser/Thr phosphorylation, inasmuch as treatment with phosphatase 2A inactivates kinase activity and increases the mobility of the bands on SDS-PAGE in an okadaic acid-sensitive manner. Thus, the recombinant p70 S6 kinase undergoes multiple phosphorylation and partial activation in COS cells. Acquisition of S6 protein kinase catalytic function, however, is apparently restricted to the most extensively phosphorylated recombinant polypeptides.
Amino Acid Sequence↗
Medulloblastoma: long-term follow-up of patients treated with electron irradiation of the spinal field.
Thirty-two patients with posterior fossa medulloblastoma underwent treatment with electron irradiation to the spinal field. The 5- and 10-year actuarial survival rates were 57% and 50%, respectively. Late complications observed in the 15 patients followed up for more than 5 years were short stature (six patients), decreased sitting-standing height ratio (four patients), scoliosis (two patients), poor school performance (seven patients), xerostomia (one patient), esophageal stricture (one patient), pituitary dysfunction (four patients), primary hypothyroidism (one patient), bilateral eighth-nerve deafness (one patient), and carcinoma of the thyroid (one patient). Complications following treatment with electrons to a spinal field are compared with reported complications following treatment with photons to the spinal field. Although short-term reactions were minimal, the authors found no difference in late complications. More sophisticated treatment planning may show such a long-term benefit in the future.
Adolescent↗