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Biomedical subjects

K Roy

Publications and source records attributed to K Roy.

At least 73 records · Page 4Linked to original sources
PubMed1999

Oral gene delivery with chitosan--DNA nanoparticles generates immunologic protection in a murine model of peanut allergy.

Food allergy is a common and often fatal disease with no effective treatment. We describe here a new immunoprophylactic strategy using oral allergen-gene immunization to modulate peanut antigen-induced murine anaphylactic responses. Oral administration of DNA nanoparticles synthesized by complexing plasmid DNA with chitosan, a natural biocompatible polysaccharide, resulted in transduced gene expression in the intestinal epithelium. Mice receiving nanoparticles containing a dominant peanut allergen gene (pCMVArah2) produced secretory IgA and serum IgG2a. Compared with non-immunized mice or mice treated with 'naked' DNA, mice immunized with nanoparticles showed a substantial reduction in allergen-induced anaphylaxis associated with reduced levels of IgE, plasma histamine and vascular leakage. These results demonstrate that oral allergen-gene immunization with chitosan-DNA nanoparticles is effective in modulating murine anaphylactic responses, and indicate its prophylactic utility in treating food allergy.

2S Albumins, Plant↗
PubMed1999

A 75-kDa Na+,K+-ATPase competitive inhibitor protein isolated from rat brain cytosol binds to a site different from the ouabain-binding site.

A Na+,K+-ATPase inhibitor protein has been purified to homogeneity from rat brain cytosol by ammonium sulphate precipitation, DEAE anion-exchange chromatography and hydroxyapatite adsorption column chromatography. The purified protein migrates as a single polypeptide band of 75 kDa on 7.5% SDS/PAGE. Amino acid composition data shows the presence of a high number of acidic amino acids in the molecule in relation to the pI value of 4.6. The inhibitor binds Na+,K+-ATPase reversibly and blocks ATP binding sites at micromolar concentrations with an I50 of approximately 700 nm. As a result, formation of the phosphorylated intermediate of Na+,K+-ATPase is hindered in the presence of the inhibitor. It does not affect p-nitrophenylphosphatase activity. Tryptophan fluorescence studies and CD analysis suggest conformational changes of Na+,K+-ATPase on binding to the inhibitor.

Amino Acids↗
PubMed1999

Sub-grouping non-melancholic major depression using both clinical and aetiological features.

OBJECTIVE: In previous papers we have considered the extent to which two contrasting analytic approaches, examining reported clinical symptom variables alone and aetiological variables alone, assist definition of subgroups of non-melancholic major depression. Here, we address the same objective but combine both sets of variables, and contrast the combined solution with each of the contributing ones. METHOD: We study a sample of 185 subjects with a putative non-melancholic major depressive disorder, with analyses involving 13 aetiological and 38 symptom variables. RESULTS: A four-class subgrouping was derived by use of a cluster analytic technique, with 'neurotic depression', non-anxious 'depressed', 'situational' and 'residual' groups. The largest group comprised 'neurotic depression' subjects, with characteristics compatible with a spectrum disorder encompassing both clinical features as well as an underlying temperament and personality style marked by anxiety. CONCLUSIONS: Comparative advantages and properties of the three differing analytic approaches to defining 'meaningful' non-melancholic major depressive subgroupings are considered. As a 'neurotic depressive' class has been consistently identified across those three approaches, but with quite varying numbers of subjects circumscribed, it is clearly a 'fuzzy' entity which may benefit from a dimensional approach to its measurement. As many of the non-melancholic groupings appear secondary to a substantive predisposing factor such as anxiety or disordered personality functioning, the clinical importance and treatment utility in identifying and circumscribing such classes are clearly supported.

Adult↗
PubMed1999

Are the newer antidepressant drugs as effective as established physical treatments? Results from an Australasian clinical panel review.

OBJECTIVE: The aim of this study was to determine, in a clinical panel sample, the extent to which patients with depression (and melancholic and non-melancholic subtypes) judged the effectiveness of previously received antidepressant treatments, particularly the comparative effectiveness of the older and newer antidepressant drugs. METHOD: Twenty-seven Australasian psychiatrists assessed 341 non-psychotic depressed patients and rated the extent to which previous antidepressant treatments had been effective. Patients were assigned to 'melancholic' and residual 'non-melancholic' categories by two processes (DSM-IV decision rules, and a cluster analysis-derived allocation) and treatment effectiveness examined within each category. RESULTS: Electroconvulsive therapy (both bilateral and unilateral) was judged as highly effective by both melancholic and non-melancholic patients. Antipsychotic medication similarly rated highly (but was judged as more effective by the non-melancholic than melancholic patients). The tricyclics and irreversible monoamine oxidase inhibitors (MAOIs) were rated as more effective by the whole sample than several newer antidepressant classes (including the selective serotonin re-uptake inhibitors [SSRIs], venlafaxine, mianserin and moclobemide), whether effectiveness was examined dimensionally or categorically. Comparison of the overall tricyclic and SSRI classes indicated that any superior tricyclic effectiveness was specific to the melancholic subjects. CONCLUSIONS: Despite methodological limitations intrinsic to such clinical panel data, the judged greater effectiveness of the older antidepressants (tricyclics and irreversible MAOIs) for melancholic depression is of importance. If valid, such data are of intrinsic clinical relevance but also have the potential to inform us about the neurobiological determinants of 'melancholia' and pharmacological actions which contribute to its effective treatment.

Antidepressive Agents↗
PubMed1999

Subtyping depression: testing algorithms and identification of a tiered model.

We seek to distinguish psychotic, melancholic, and nonmelancholic depression by clinical features and to test varying algorithm models to determine optimal criteria sets. We report a study of 269 depressed inpatients and outpatients. A latent class analysis (LCA) of 16 clinical features allowed for specificity or overrepresentation of features to be examined across the three classes. Varying algorithm models for distinguishing melancholic and nonmelancholic depression, involving endogeneity symptoms and observer-rated psychomotor disturbance (PMD) were compared. Psychotic depression was readily distinguished by the specific presence of psychotic features, and PMD was most severe in this class. Melancholic depression was most clearly distinguished from the residual nonmelancholic class by the presence of PMD. Although some endogeneity symptoms were overrepresented in the melancholic class, their specificity was unimpressive. An algorithm involving PMD components alone was highly efficient in discriminating LCA classes and, more importantly, superior to DSM-IV decision rules when examined against a range of clinical validators of melancholia. Subtyping appears assisted by a hierarchical model, based on a small set of features. The move from nonmelancholic to melancholic depression appears defined by a tier of observably rated PMD, whereas the move from melancholic to psychotic depression is determined by a tier of psychotic features and contributed to by significantly higher levels of PMD.

Adult↗
PubMed1999

Hepatitis C virus and oral disease: a critical review.

Hepatitis C virus (HCV) infection is widespread with an estimated 3% of the world population being infected. Acute infection is usually mild but chronicity develops in as many as 70% of patients, of whom at least 20% will eventually develop cirrhosis. A further 1-4% of cirrhotic individuals will develop hepatocellular carcinoma. Infection with HCV may have effects on various organs other than the liver. HCV has been causally associated with a remarkable array of extrahepatic manifestations, some of which remain unproven. This review discusses the evidence implicating HCV in the aetiology of two important oral conditions, namely Sjögren's syndrome and lichen planus.

Hepacivirus↗
PubMed1999

Delayed cell death, giant cell formation and chromosome instability induced by X-irradiation in human embryo cells.

We studied X-ray-induced delayed cell death, delayed giant cell formation and delayed chromosome aberrations in normal human embryo cells to explore the relationship between initial radiation damage and delayed effect appeared at 14 to 55 population doubling numbers (PDNs) after X-irradiation. The delayed effect was induced in the progeny of X-ray survivors in a dose-dependent manner and recovered with increasing PDNs after X-irradiation. Delayed plating for 24 h post-irradiation reduced both acute and delayed lethal damage, suggesting that potentially lethal damage repair (PLDR) can be effective for relieving the delayed cell death. The chromosome analysis revealed that most of the dicentrics (more than 90%) observed in the progeny of X-ray survivors were not accompanied with fragments, in contrast with those observed in the first mitosis after X-irradiation. The present results indicate that the potentiality of genetic instability is determined during the repair process of initial radiation damage and suggest that the mechanism for formation of delayed chromosome aberrations by radiation might be different from that of direct radiation-induced chromosome aberrations.

Cell Death↗
PubMed1999

An exploration of links between early parenting experiences and personality disorder type and disordered personality functioning.

Reports of early parenting were assessed using two measures, the Parental Bonding Index (PBI) and the Measure of Parenting Style (MOPS), in a sample of 265 patients with DSM-defined major depressive disorder. Psychiatrists then rated the extent to which sample members evidenced the personality "styles" underpinning 15 separate personality disorders, returning personality vignette scores. The extent of disordered functioning was also assessed across "parameters" and "domains" by psychiatrists, referrers, and family members, using a range of measures. Those with higher scores on vignettes measuring borderline, anxious, depressive, and self-defeating personality style rated parents as uncaring, overcontrolling, and abusive. When vignettes were consolidated into scores akin to the DSM clusters, the most consistent links between perceived dysfunctional parenting were with the Cluster C (anxious), and Cluster B (dramatic) styles and were nonsignificant for Cluster A (eccentric) style. Meeting criteria for an increasing number of personality disorder clusters was associated with increasing levels of adverse parenting. Multiple regression analyses indicated that disordered functioning (as assessed by the three independent rater groups) was most distinctly associated with paternal indifference and maternal overcontrol.

Adult↗
PubMed1999

Analysis of two cosmid clones from chromosome 4 of Drosophila melanogaster reveals two new genes amid an unusual arrangement of repeated sequences.

Chromosome 4 from Drosophila melanogaster has several unusual features that distinguish it from the other chromosomes. These include a diffuse appearance in salivary gland polytene chromosomes, an absence of recombination, and the variegated expression of P-element transgenes. As part of a larger project to understand these properties, we are assembling a physical map of this chromosome. Here we report the sequence of two cosmids representing approximately 5% of the polytenized region. Both cosmid clones contain numerous repeated DNA sequences, as identified by cross hybridization with labeled genomic DNA, BLAST searches, and dot matrix analysis, which are positioned between and within the transcribed sequences. The repetitive sequences include three copies of the mobile element Hoppel, one copy of the mobile element HB, and 18 DINE repeats. DINE is a novel, short repeated sequence dispersed throughout both cosmid sequences. One cosmid includes the previously described cubitus interruptus (ci) gene and two new genes: that a gene with a predicted amino acid sequence similar to ribosomal protein S3a which is consistent with the Minute(4)101 locus thought to be in the region, and a novel member of the protein family that includes plexin and met-hepatocyte growth factor receptor. The other cosmid contains only the two short 5'-most exons from the zinc-finger-homolog-2 (zfh-2) gene. This is the first extensive sequence analysis of noncoding DNA from chromosome 4. The distribution of the various repeats suggests its organization is similar to the beta-heterochromatic regions near the base of the major chromosome arms. Such a pattern may account for the diffuse banding of the polytene chromosome 4 and the variegation of many P-element transgenes on the chromosome.

Amino Acid Sequence↗
PubMed1999

Assessment of the need for a training course in rational use of drugs: a cross-sectional pilot study.

Rational use of drugs means need-based use of them keeping in mind the pathological status, therapeutic indices, drugs interactions and adverse drug reactions. Lacunae in the existing undergraduate curriculum cause irrational use of drugs. The present study was undertaken among 2,200 fresh medical graduates from all over the country with a list of questionnaire distributed among them and analysing the answers. The aim was to ascertain the adequacy of present undergraduate curriculum on pharmacology in equipping the doctors on rational use of drugs and to assess the need and feasibility of Refresher's course. More than half (55%) replied prescriptions were not truly rational. Ninety-eight per cent opened Refresher's course is beneficial in rational prescribing. This cross-sectional survey could provide a glimpse of existing undergraduate pharmacology curriculum and its impact on rational prescribing practice. The Refresher's course in the early internship period involving the clinical departments and department of pharmacology is suggested.

Cross-Sectional Studies↗
PubMed1998

Cloning of mouse gamma-glutamyl hydrolase in the form of two cDNA variants with different 5' ends and encoding alternate leader peptide sequences.

Mouse-liver gamma-glutamyl hydrolase (GH) is a lysosomal endopeptidase with an acid pH optimum that is activated by sulfhydryl compounds and preferentially hydrolyzes the most proximal gamma-glutamyl linkage of longer chain polyglutamates of folates and their analogues. We describe the cloning of this mouse lysosomal cDNA enzyme from liver GH mRNA in the form of two cDNA variants (1.295 and 1.268 kb in length) differing 14-fold (Variant I versus Variant II) in relative frequency that exhibited 5'-end heterogeneity and encoded alternate leader peptides. The 5' UTR in these variants also differs in length by 27 nucleotides. Otherwise, the ORF and 3' UTR in each case are the same. These cDNAs encode a protein in which the deduced amino acid sequence shares 78.9 and 69. 1% identity to rat and human GH sequences, respectively. Amino acid sequence comparisons among the three species identified three conserved Asn sites and two conserved Cys residues that may be sites of glycosylation and sulfhydryl compound activation, respectively. Variant I GH mRNA was more abundant than Variant II GH mRNA in all mouse tissues examined. Variant I GH mRNA levels were extremely high in salivary gland, moderately high in kidney, liver, lung, stomach and uterus, low in small intestine, brain and fetal liver and relatively rare in thymus, spleen and skeletal muscle. Abundance of GH mRNA among tumors varied from low to high, with no discernible correlation with their tissue of origin.

Amino Acid Sequence↗
PubMed1998

Structural analysis of the human RFC-1 gene encoding a folate transporter reveals multiple promoters and alternatively spliced transcripts with 5' end heterogeneity.

The organization and structure of the human RFC-1 gene encoding a folate transporter were determined. The RFC-1 gene spans 22.5kb and was found to be distributed in eight exons, including five primary exons and three alternatives of exon 1. Most splice junctions conform to consensus sequences for such junctions. The human RFC-1 gene differs from the mouse and hamster genes both in terms of the total number of exons and in regard to alternatives of exon 1 which encode 5' end heterogeneity. Previously described cDNA variants (GenBank/EMBL accession no. U19720) are now shown to incorporate one of two alternatives (exons 1a and 1b) to exon 1 and exons 2-6 as a result of RNA splicing. Another variant also described may not be full length in that it incorporates a probable alternative (exon 1c) to exon 1 along with exon 2 and a truncated exon 3. A relatively GC- rich region of the genome 5' of the alternatives to exon 1 appears to be distinctly promoter like and incorporates a number of putative cis-acting elements, including multiple SP1 sites, involved in the regulation of transcription. Primer extension analysis of this upstream region in two human cell types revealed a similar pattern of multiple transcription start sites (tsp) proximal to the 5' end of exon 1. However, there was a greater number of potential tsp within the region immediately upstream of exon 1b than within the regions upstream of exons 1a and 1c. The existence of true alternatives to exon 1 in this gene incorporating different 5' ends indicates that its transcription is under the control of multiple promoters. The identity of two such promoters was obtained by functional deletion analysis, showing that expression of a luciferase reporter gene was directed separately by discrete stretches of nucleotide sequence proximal to exon 1a (promoter 1) or exon 1b (promoter 2) in transient transfection experiments. Promoter 1 appeared to have a three-fold lower basal activity than promoter 2, but was enhanced up to nine-fold in fusion constructs containing an SV40 enhancer element. Also, promoter 2 partly consists of a highly GC-rich direct repeat element containing at least three putative SP-1 and 3 putative MZF1 sites. Finally, the activity of these promoters relative to each other was consistent with the results of primer extension analysis showing a greater multiple and usage of tsp within promoter 2 (exon 1b) than within promoter 1 (exons 1a and 1c), suggesting that the variant incorporating exon 1b was the most abundant.

Alternative Splicing↗
PubMed1998

DNA-polycation nanospheres as non-viral gene delivery vehicles.

Nanospheres synthesized by salt-induced complex coacervation of cDNA and polycations such as gelatin and chitosan were evaluated as gene delivery vehicles. DNA-nanospheres in the size range of 200-750 nm could transfect a variety of cell lines. Although the transfection efficiency of the nanospheres was typically lower than that of lipofectamine and calcium phosphate controls in cell culture, the beta-gal expression in muscle of BALB/c mice was higher and more sustained than that achieved by naked DNA and lipofectamine complexes. This gene delivery system has several attractive features: (1) ligands can be conjugated to the nanosphere for targeting or stimulating receptor-mediated endocytosis; (2) lysosomolytic agents can be incorporated to reduce degradation of the DNA in the endosomal and lysosomal compartments; (3) other bioactive agents or multiple plasmids can be co-encapsulated; (4) bioavailability of the DNA can be improved because of protection from serum nuclease degradation by the polymeric matrix; (5) the nanosphere can be lyophilized for storage without loss of bioactivity.

Animals↗
PubMed1998

Marine latitudinal diversity gradients: tests of causal hypotheses.

Latitudinal diversity gradients are first-order expressions of diversity patterns both on land and in the oceans, although the current hypotheses that seek to explain them are based chiefly on terrestrial data. We have assembled a database of the geographic ranges of 3,916 species of marine prosobranch gastropods living on the shelves of the western Atlantic and eastern Pacific Oceans, from the tropics to the Arctic Ocean. Western Atlantic and eastern Pacific diversities are similar, and the diversity gradients are strikingly similar despite many important physical and historical differences between the oceans. This shared diversity pattern cannot be explained by: (i) latitudinal differences in species range-length (Rapoport's rule); (ii) species-area effects; or (iii) recent geologic histories. One parameter that does correlate significantly with diversity in both oceans is solar energy input, as represented by average sea surface temperature. If this correlation is causal, sea surface temperature is probably linked to diversity through some aspect of productivity. In this case, diversity is an evolutionary outcome of trophodynamic processes inherent in ecosystems, and not just a byproduct of physical geographies.

Animals↗
PubMed1998

A single amino acid difference within the folate transporter encoded by the murine RFC-1 gene selectively alters its interaction with folate analogues. Implications for intrinsic antifolate resistance and directional orientation of the transporter within the plasma membrane of tumor cells.

The apparent Km, but not Vmax, for influx of methotrexate (MTX) mediated through the plasma membrane of S180 cells by the one-carbon, reduced folate transporter as well as the KD for binding to the transporter were 4-fold higher than in L1210 cells correlating with the greater intrinsic resistance of the former to this folate analogue. In contrast, no difference was observed between each cell type with regard to efflux of [3H]MTX mediated by this same transporter in ATP-depleted cells. The difference in influx Km in the case of this 10-methyl substituted N1O analogue of folic acid was not seen with more effective permeants, such as the unsubstituted N1O aminopterin or C1O analogues. Thus, values for influx Km for aminopterin, which were 1-1.2 microM in each cell type, increased as a result of substitution at N1O (MTX) 3-fold in L1210 cells but 12-fold in S180 cells. Nucleotide sequencing of reverse transcriptase-polymerase chain reaction-generated cDNA and of polymerase chain reaction-generated genomic DNA identified a single nucleotide difference between each cell type at +890 within exon 3 of the RFC-1 gene. This was in the form of a G (L1210 cells) to A (S180 cells) transition. Codon 297, the site of this transition, encodes either Ser or Asn in L1210 or S180 cells, respectively, which is located between the seventh and eight membrane-spanning helices. This amino acid difference had no effect on the electrophoretic mobility or amount of the transporter in each cell type that was shown by Western blotting with anti-RFC-1 peptide antibodies to migrate as 46 kDa in each case. Proof that this nucleotide difference alone accounted for the alteration in influx between each cell type was obtained by S180 RFC-1 cDNA versus L1210 RFC-1 cDNA transfection of an L1210 cell variant with undetectable MTX influx and RFC-1 gene expression. In this case, the higher Km for MTX influx associated with S180 cells was duplicated only in the S180 RFC-1 transfectants. These results appear to document the first example of a nucleotide alteration within the RFC-1 gene, which influences the interaction of MTX with the encoded plasma membrane transporter. An analysis of topology, in addition to other considerations, suggests that the site of the amino acid difference found in the transporter from L1210 and S180 cells occurs within or near the binding site on the external plasma membrane surface.

Amino Acid Sequence↗
PubMed1998

Chromosomal localization of the murine RFC-1 gene encoding a folate transporter and its amplification in an antifolate resistant variant overproducing the transporter.

A variant of the L1210 cell (L1210/R83) selected in the presence of the lipophilic antifolate, metoprine, and a concentration of the natural diastereoisomer of 5-formyltetrahydrofolate, lL5CHO-folateH4, suboptimum for growth exhibited a 35-fold increase compared to parental L1210 cells in one-carbon, reduced folate transport. This was evidenced by the increase in Vmax for [3H]MTX (methotrexate) influx and a commensurate increase in the amount of the 46 kilodalton (kDa) transport protein and reduced folate carrier (RFC-1) mRNA. The variant is resistant to lipophilic antifolates, but shows collateral sensitivity to classical folate analogues. Karyotype analysis of L1210/R83 cells revealed the presence of several new chromosome abnormalities. One of these was a large, submetacentric marker chromosome comprising a normal #10 and a longer, abnormally banded arm of uncertain origin which exhibited an interstitial, palely staining, HSR-like segment. The results of Southern and Northern blotting showed that the RFC-1 gene copy number and RNA transcript level were markedly increased (30-35 fold) in L1210/R83 cells. Fluorescence in situ hybridization (FISH) analysis revealed that the HSR-like segment in these cells was the site of amplified RFC-1 genes. Independent revertant subclones, obtained following growth in the absence of selection pressure, showed four- to 12-fold decreases in [3H]MTX influx Vmax and in amount of NHS (N-hydroxysuccinimide)-[3H]MTX affinity labeled one-carbon, reduced folate transporter compared to L1210/R83 cells. RFC-1 gene copy number also decreased, and the mean length of the HSR in these revertants declined 1.6- to 5-fold. Based upon genomic nucleotide sequencing, the RFC-1 gene in the normal mouse genome was localized to chromosome 10 in close association with the alpha 1 (Col18a1) collagen gene at 10B3(locus 41cM). The close association of these genes was confirmed by other data showing that the alpha 1 collagen gene was co-amplified in L1210/R83 cells. These results document the amplification at the site of a putative HSR in an L1210 cell variant of the RFC-1 gene regulating expression of the one-carbon, reduced folate transporter.

Animals↗
PubMed1998

"Acting out" and "acting in" as behavioral responses to stress: a qualitative and quantitative study.

In a sample of 270 depressed patients, we describe some behaviors in response to stress. One third acknowledged "acting out" behaviors--angry, destructive acts and "out of control" behaviors. Four fifths acknowledged "acting in" behaviors--most commonly withdrawal. As a percentage of subjects acknowledged both response styles (with alternate expression influenced by situation), we developed a dimensional self-report measure within a subsample of 177 who attended a follow-up reassessment. Assignment to four groups with contrasting expressions of "acting out" and "acting in" scores demonstrated differences in age, diagnostic status, age of onset of depression, and self-injurious behaviors.

Acting Out↗