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S K Arora

Publications and source records attributed to S K Arora.

At least 37 records · Page 2Linked to original sources
PubMed2000

fleN, a gene that regulates flagellar number in Pseudomonas aeruginosa.

The single polar flagellum of Pseudomonas aeruginosa plays an important role in the pathogenesis of infection by this organism. However, regulation of the assembly of this organelle has not been delineated. In analyzing the sequence available at the Pseudomonas genome database, an open reading frame (ORF), flanked by flagellar genes flhF and fliA, that coded for a protein (280 amino acids) with an ATP-binding motif at its N terminus was found. The ORF was inactivated by inserting a gentamicin cassette in P. aeruginosa PAK and PAO1. The resulting mutants were nonmotile on motility agar plates, but under a light microscope they exhibited random movement and tumbling behavior. Electron microscopic studies of the wild-type and mutant strains revealed that the mutants were multiflagellate, with three to six polar flagella per bacterium as rather than one as in the wild type, indicating that this ORF was involved in regulating the number of flagella and chemotactic motility in P. aeruginosa. The ORF was named fleN. An intact copy of fleN on a plasmid complemented the mutant by restoring motility and monoflagellate status. The beta-galactosidase activities of eight flagellar operon or gene promoters in the wild-type and fleN mutant strains revealed a direct correlation between six promoters that were upregulated in the fleN mutant (fliLMNOPQ, flgBCDE, fliEFG, fliDS orf126, fleSR, and fliC) and positive regulation by FleQ, an NtrC-like transcriptional regulator for flagellar genes. Based on these results, we propose a model where FleN influences FleQ activity (directly or indirectly) in regulating flagellar number in P. aeruginosa.

Amino Acid Sequence↗
PubMed1999

Synthesis and antitumor activity of goniofufurone analogues.

Synthesis and antitumor activity of goniofufurone analogues 15, 16, 17, 33, and 46 is reported. Key step in the synthesis is Pd (II) mediated oxidative cyclisation of vinyl-(hydroxy) furans 18, 19 to the corresponding lactols 32, 43. Cytotoxicities of 15, 16, 17, 33, and 46 tested against six human cancer cell lines are reported. Change of stereochemistry at C-5, C-6 and C-7 position of goniofufurone (1) did not enhance the cytotoxicities significantly.

Antineoplastic Agents↗
PubMed1999

Diagnostic efficacy of polymerase chain reaction in granulomatous uveitis.

SETTING: The granulomatous uveitis, multifocal choroiditis and periphlebitis have been suspected to be of tubercular origin but no definitive reports about detection of etiological agents have been documented in the literature. Conventional bacteriological methods are not generally helpful in diagnosing ocular tuberculosis due to difficulty with potential morbidity associated with obtaining the biopsy material from the eye. Thus, the diagnosis of ocular tuberculosis is most often presumptive. OBJECTIVE: We evaluated the role of polymerase chain reaction (PCR) for detection of Mycobacterium tuberculosis in the aqueous humor samples obtained from eyes with active uveitis. METHODS: Aqueous samples from 53 patients having cellular reaction in the anterior chamber along with any one or more of the following: 1) active vasculitis; 2) anterior vitreous cells; 3) snowball opacities; 4) snow banking in the pars plana; 5) retinochoroiditis were withdrawn by anterior chamber paracentesis and subjected to PCR. Seventeen samples from patients with definite clinical diagnoses other than tuberculosis formed a disease control group. Fifteen aqueous samples obtained from healthy subjects undergoing routine cataract surgery served as healthy controls. PCR was performed using primers capable of amplifying a 150 b.p. segment from a conserved repetitive sequence in the genome of M. tuberculosis. RESULTS: Twenty out of the 53 samples (37.7%) in the study group were positive where as only one sample out of 17 in the disease control group (5.7%) showed a weakly positive band. No sample from the healthy control group showed a positive PCR. CONCLUSION: Our study shows that PCR can be effectively used for the diagnosis of intraocular tuberculosis in the presence of uveitis.

Aqueous Humor↗
PubMed1998

Genetic polymorphism of Leishmania species using kinetoplast DNA restriction fragment length polymorphism and cDNA probe of Leishmania donovani.

Leishmaniasis represents a group of diseases that range from simple cutaneous lesions through metastasizing diffused cutaneous to severe systemic infection depending upon the taxon to which the causative parasite belongs. Therefore, it is important to identify the infecting Leishmania. Methods presently being used, including immunology, biochemistry and molecular biology have one or the other limitations, leaving scope for the search for newer probes. This study reports the characterization of leishmania isolates both by restriction fragment length polymorphism of kinetoplast DNA (kDNA) and genomic DNA. The genomic DNA was probed with a cDNA probe B2a1. Using a kDNA restriction pattern technique, different isolates of Leishmania donovani could be differentiated from the UR6 strain of L. tropica, but it was not possible to differentiate between newer local isolates of L. donovani with most of the restriction enzymes except AluI. However, the B2a1 cDNA probe was able to differentiate these isolates effectively. Both of these techniques could differentiate newer local isolates of L. donovani from the older isolates of L. donovani from India, i.e., DD8, RMRI and SS. The Indian isolates of L. donovani could also be differentiated from isolates of L. donovani from Jeddah and Germany using both techniques. The present study indicates that the cDNA probe B2a1 can be used as an important adjunct to kDNA restriction analysis for the characterization of Leishmania species.

Animals↗
PubMed1998

Heterogeneity in heat shock protein genes in Leishmania isolates.

Leishmaniasis, a group of visceral and cutaneous diseases, is caused by parasites belonging to one genus comprising approximately 13 different species. Many methods including serological, biochemical and molecular biological techniques have been used by various workers to characterize these different species and isolates of Leishmania, yet there is no single generally accepted criterion. We have identified certain cDNA clones from a library generated from the promastigotes of S1 strain of Leishmania donovani and used them as probes for identification of various isolates of L. donovani and Leishmania tropica. Two of the probes used, E2b (2.0 kb) and E1a (1.3 kb), sequence characterized to be hsp70 of Leishmania, were able to distinguish various isolates of L. donovani from different geographical origins as well as strains of L. donovani from those of L. tropica. Thus, by using recombinant hsp70 cDNA probes, the data indicated that there is a considerable degree of heterogeneity in the heat-shock genes of Leishmania.

Animals↗
PubMed1998

The Pseudomonas aeruginosa flagellar cap protein, FliD, is responsible for mucin adhesion.

Mucin-specific adhesion of Pseudomonas aeruginosa plays an important role in the initial colonization of this organism in the airways of cystic fibrosis patients. We report here that the flagellar cap protein, FliD, participates in this adhesion process. A polar chromosomal insertional mutation in the P. aeruginosa fliD gene made this organism nonadhesive to mucin in an in vitro mucin adhesion assay. The adhesive phenotype was restored by providing the fliD gene alone on a multicopy plasmid, suggesting involvement of this gene in mucin adhesion of P. aeruginosa. Further supporting this observation, the in vitro competition experiments demonstrated that purified FliD protein inhibited the mucin adhesion of nonpiliated P. aeruginosa PAK-NP, while the same concentrations of PilA and FlaG proteins of P. aeruginosa were ineffective in this function. The regulation of the fliD gene was studied and was found to be unique in that the transcription of the fliD gene was independent of the flagellar sigma factor sigma28. Consistent with this finding, no sigma28 binding sequence could be identified in the fliD promoter region. The results of the beta-galactosidase assays suggest that the fliD gene in P. aeruginosa is regulated by the newly described transcriptional regulator FleQ and the alternate sigma factor sigma54 (RpoN).

Amino Acid Sequence↗
PubMed1997

A transcriptional activator, FleQ, regulates mucin adhesion and flagellar gene expression in Pseudomonas aeruginosa in a cascade manner.

Previous work has demonstrated that fleR, the gene for a transcriptional activator belonging to the NtrC subfamily of response regulators, is involved in the regulation of mucin adhesion and flagellar expression by Pseudomonas aeruginosa. This report describes the identification and characterization of fleQ, the gene for another transcriptional regulator which also regulates mucin adhesion and motility in this organism. The complete nucleotide sequence of the fleQ gene was determined on both DNA strands, and an open reading frame (ORF) consisting of 1,493 nucleotides was identified. This ORF coded for a gene product of predicted molecular weight, as confirmed by the overexpression of the fleQ gene as a fusion protein under an inducible promoter. The fleQ gene is flanked by a flagellar operon, fliDSorf126, at the 5' end and the fleSR operon on the 3' end. FleQ also had striking homology to a number of proteins belonging to the NtrC subfamily of response regulators, which work in concert with the alternate sigma factor RpoN (sigma54) to activate transcription. However, FleQ lacks the residues corresponding to Asp-54 and Lys-104 of the NtrC protein which are conserved in most of the members belonging to this subfamily of regulators. In addition, unlike some of the other transcriptional activators of this group, FleQ does not appear to have a cognate sensor kinase. A chromosomal insertional mutation in the fleQ gene abolished mucin adhesion and motility of P. aeruginosa PAK and PAK-NP. Both of these functions were regained by providing the complete fleQ gene on a multicopy plasmid. The location of fleQ immediately upstream of the fleSR operon, which is also necessary for the same process, suggested that these regulators may interact in some way. We therefore examined the regulation of the fleSR operon by fleQ and vice versa. Promoter fusion experiments showed that the fleSR operon was regulated by RpoN and FleQ. On the other hand, the fleQ promoter was independent of RpoN and FleR. FleQ, thus, adds another level of regulation to motility and adhesion in P. aeruginosa, above that of fleSR. We therefore propose the existence of a regulatory cascade which consists of at least two transcriptional regulators, FleQ and FleR, in the control of motility and adhesion in P. aeruginosa.

Amino Acid Sequence↗
PubMed1996

Convulsions in tuberculous meningitis.

One-hundred-and-thirty-six children below 12 years of age hospitalized with a diagnosis of tuberculous meningitis (TBM) have been investigated to identify the underlying cause of convulsions. One-hundred-and-one children (74 per cent) presented with seizures before and/or during hospitalization. Generalized tonic and clonic seizures (GTCS) were the commonest (58 per cent) type of seizures followed by focal seizures (FS) (38 per cent) and tonic spasms (TS) (4 per cent). EEG changes were more frequently observed in cases with FS and in those children with GTCS who presented after first week of hospitalization. EEG findings included generalized dysrythmia with paroxysmal slow activity (38 per cent), interhemispheric asymmetry (23 per cent), multiple spike and wave pattern (10 per cent), and focal spike and wave pattern (15 per cent). CT scan findings were more common in those children with GTCS and TS who presented with recurrent seizures and/or seizures manifesting after first week of hospitalization. FS presenting at any stage of the disease were associated with CT scan abnormalities. Abnormalities detected in CT scan of brain included meningeal enhancement (55 per cent), hydrocephalus (32 per cent), tuberculomas (27 per cent), and cerebral infarctions (13 per cent). Clinical presentation and investigations indicate that the probable cause of convulsions could be attributed to cerebral edema (57 per cent), syndrome of inappropriate secretion of antidiuretic hormone (35 per cent), hydrocephalus (32 per cent), tuberculoma (27 per cent), abnormal electric focus (25 per cent), and cerebral infarction (13 per cent).

Anticonvulsants↗
PubMed1996

Cloning and characterization of Pseudomonas aeruginosa fliF, necessary for flagellar assembly and bacterial adherence to mucin.

Pseudomonas aeruginosa adheres to the mucosal surfaces of the lungs. This process appears to be mediated by nonpilus adhesins which bind to mucin. To find this nonpilus adhesin(s), mutagenesis of a nonpiliated mutant of P. aeruginosa with transposon Tn5G, followed by a screen for mucin adhesion, was used to isolate a series of mutants unable to adhere to mucin. All of these mutants were also found to be defective in motility. One such mutant, PAK-RR20, is characterized here. The site of the transposon insertion in PAK-RR20 was localized to a gene which is homologous to the fliF gene of other organisms and was flanked by other motility-related genes, fliE and fliG. Both adhesion and motility defects in PAK-RR20 were complemented by providing the fliF gene in trans. Since complementation could have been due to the presence of an internal promoter in the fliF gene or in the Tn5G transposon, which allowed the transcription of the downstream genes, another chromosomal mutant of the fliF gene was constructed by insertional inactivation with an antibiotic resistance cassette. This mutant was also nonmotile and nonadhesive. However, the two defects in this new mutant could not be complemented by the fliF gene in trans, consistent with the interpretation that there is no internal fliF promoter but possibly a functional promoter in the Tn5G transposon. The complete nucleotide sequences of the fliE and fliF genes and a partial nucleotide sequence of the fliG gene of P. aeruginosa were determined. Control of the promoter upstream of the fliE gene was analyzed by construction of a fliE-lacZ fusion and the introduction of this construct into strains of P. aeruginosa with mutations in several regulatory genes. Beta-Galactosidase expression measurements indicated that the fliE promoter does not utilize RpoF (sigma(28)) or RpoN (sigma(54)) sigma factors. The characterization of this gene as being responsible for the loss of adhesion indicates that basal body structures are probably important for localization of the adhesin.

Amino Acid Sequence↗
PubMed1996

The nucleotide sequence of the Pseudomonas aeruginosa pyrE-crc-rph region and the purification of the crc gene product.

The gene (crc) responsible for catabolite repression control in Pseudomonas aeruginosa has been cloned and sequenced. Flanking the crc gene are genes encoding orotate phosphoribosyl transferase (pyrE) and RNase PH (rph). New crc mutants were constructed by disruption of the wild-type crc gene. The crc gene encodes an open reading frame of 259 amino acids with homology to the apurinic/apyrimidinic endonuclease family of DNA repair enzymes. However, crc mutants do not have a DNA repair phenotype, nor can the crc gene complement Escherichia coli DNA repair-deficient strains. The crc gene product was overexpressed in both P. aeruginosa and in E. coli, and the Crc protein was purified from both. The purified Crc proteins show neither apurinic/apyrimidinic endonuclease nor exonuclease activity. Antibody to the purified Crc protein reacted with proteins of similar size in crude extracts from Pseudomonas putida and Pseudomonas fluorescens, suggesting a common mechanism of catabolite repression in these three species.

Amino Acid Sequence↗
PubMed1996

Recognition of mucin by the adhesin-flagellar system of Pseudomonas aeruginosa.

Pseudomonas aeruginosa colonizes the mucus of patients with chronic lung diseases by a specific mechanism involving an adhesin-receptor system. Several adhesins have been implicated in the adhesion of P. aeruginosa to cells, but the identity of the principal adhesin(s) involved in adhesion to mucin is unknown. Mutagenesis studies have indicated that P. aeruginosa adhesion is under the control of the rpoN gene, which also regulates pilin synthesis, flagellum formation, and other functions. Mutagenesis of certain flagellar genes that are not controlled by RpoN, e.g., flif, also indicates a close relationship between adhesion and flagellar genes and not necessarily an independent effect of rpoN on adhesion. Mutants of certain early flagellar genes lead to the loss of both adhesion and motility, whereas mutants of certain late genes, e.g., fliC, the gene for flagellin, lose motility but retain adhesion. Recent studies indicate that both motility and adhesion are regulated by a two-component regulatory system called fleS-R, which in turn is controlled by another regulator in a cascade that involves rpoN. A fleR mutant possessing pili adheres poorly to mucins, definitively showing that pili do not play a major role in adhesion to mucin. It is unclear whether the adhesin is a flagellar protein or another protein that uses the flagellar export apparatus for localization or both. Finding the gene under control of rpoN may provide answers to these questions.

Adhesins, Bacterial↗
PubMed1995

The effect of Cd2+ on lipid components of sunflower (Helianthus annuus L.) seeds.

The effect of seven levels of Cd2+ viz. 0, 10, 20, 30, 40, 60 and 80 ppm (micrograms/g soil) supplied as cadmium chloride, on lipid components of sunflower seeds was studied in a pot experiment. The total and neutral lipids decreased while polar lipids (phospho and glycolipids) increased with the increasing levels of Cd2+. Oleic acid and linoleic acids were the major fatty acids in the total, polar and neutral lipid fractions. Oleic acid generally increased while linoleic acid decreased in all the lipid fractions with Cd2+ levels. The neutral lipid fraction had higher levels of palmitic, palmitoleic, stearic and oleic acid as compared to the total and polar lipid fractions. The plant dry weight and seed yield decreased whereas cadmium concentration in seeds increased consistently with increasing Cd2+ levels.

Cadmium↗
PubMed1995

Lack of serological specificity of recombinant heat shock protein of Leishmania donovani.

In order to identify a specific recombinant antigen of Leishmania donovani with potential use for diagnosis, a cDNA library was constructed in lambda ZAP II expression vector. On screening the cDNA library using pooled sera from Indian patients with kala azar, 20 antibody reactive clones were identified. These were subcloned into pBluescript phagemid by an in vivo excision procedure. The molecular weights of the expressed recombinant proteins varied from 15 to 70 kDa and the cDNA insert sizes varied from 0.5 kb to the largest size of approximately 2.0 kb which was designated as the E2b clone. The nucleotide sequencing revealed that 50% of the clones had sequence homology to the heat shock protein gene of L. donovani. The serological studies conducted with the kala azar positive sera and sera from healthy laboratory workers using the recombinant protein from the E2b clone and having sequence homology to Ldhsp 70, indicated that although all the kala azar sera was positive, 12 of 20 healthy individuals also showed antibodies against the recombinant hsp70, indicating that this antigen is not suitable for serological diagnosis of kala azar.

Animals↗
PubMed1995

Comparative evaluation of anti-heat shock protein antibodies in SLE and healthy controls.

Presence of anti-heat shock protein (anti-hsp) antibodies have earlier been demonstrated in children as well as adult patients with systemic lupus erythmatosus (SLE). Since hsp share homology across several species, it is presumed that anti-E. coli antibodies could in fact be autoantibodies. Surface expression of hsps has also been shown to play a role in the generation of antibodies. In the present study antibodies to a recombinant hsp70 of Leishmania dnonvani origin have been investigated in patients with SLE and data compared with the incidence in control sera. One clone from the cDNA library of Leishmania donovani confirmed previously to be expressing hsp70, cloned in pBluescript and antibodies to the expressed protein were checked by ELISA and western blot assays. The results indicate the presence of anti-hsp70 antibodies in 22/26 (85%) SLE patients and 12/20 (60%) control sera. Our observations reconfirm the presence of antibodies to hsp in normal controls and therefore they may not bear any direct relation to the pathogenesis of SLE.

Adult↗
PubMed1994

Changes in the anti-nutritional factors of developing seeds and pod walls of fenugreek (Trigonella foenum graecum L.).

Anti-nutritional factors of developing seeds and pod walls of fenugreek were evaluated which affect their nutritional value. Sucrose, raffinose and stachyose decreased with maturity of seeds in all the cultivars. Total, non-reducing sugars increased and reducing sugars decreased at maturity in all the cultivars. The reducing sugars decreased with maturity of pod walls. The flatus producing sugars were maximum in PEB pod walls. Saponin content increased towards maturity of seeds but decreased at maturity of seeds and decreased at maturity in pod walls of all cultivars. Phytate increased with seed maturity up to 95 days after anthesis, while phytate decreased in pod walls of all the cultivars with maturity. Total phenol, catechol and flavonol decreased with advancement of pod wall development. Total phenol decreased except HM 46 a maturity of seeds.

Catechols↗
PubMed1994

Acetylcholinesterase and Na+,K(+)-ATPase activities in different regions of rat brain during insulin-induced hypoglycemia.

The activities of acetylcholinesterase (acetylcholine acetylhydrolase, EC 3.1.1.7), responsible for hydrolysis of acetylcholine and Na+,K(+)-ATPase (Mg(2+)-dependent ATP phosphohydrolase, EC 3.6.1.3), which plays a crucial role in neurotransmission, were determined in four brain regions after 1, 2, and 3 h of insulin administration. Significant decrease in the acetylcholinesterase and Na+,K(+)-ATPase activities was observed in the soluble and total particulate fractions from cerebral hemispheres, cerebellum, brain stem, and diencephalon + basal ganglia after 1, 2, and 3 h of insulin-induced hypoglycemia. Blood glucose level decreased significantly after 1 h of insulin administration and remained at low level for 2 h thereafter, whereas, the protein content in different subcellular fractions from four brain regions did not show any significant change under this physiological stress.

Acetylcholinesterase↗