Biomedical subjects

S Subrahmanyam

Publications and source records attributed to S Subrahmanyam.

17 recordsLinked to original sources

PubMed2001

"Bite-and-Switch" approach using computationally designed molecularly imprinted polymers for sensing of creatinine.

A method for the selective detection of creatinine is reported, which is based on the reaction between polymerised hemithioacetal, formed by allyl mercaptan, o-phthalic aldehyde, and primary amine leading to the formation of fluorescent isoindole complex. This method has been demonstrated previously for the detection of creatine using creatine-imprinted molecularly imprinted polymers (MIPs) Since MIPs created using traditional methods were unable to differentiate between creatine and creatinine, a new approach to the rational design of a molecularly imprinted polymer (MIP) selective for creatinine was developed using computer simulation. A virtual library of functional monomers was assigned and screened against the target molecule, creatinine, using molecular modelling software. The monomers giving the highest binding score were further tested using simulated annealing in order to mimic the complexation of the functional monomers with template in the monomer mixture. The result of this simulation gave an optimised MIP composition. The computationally designed polymer demonstrated superior selectivity in comparison to the polymer prepared using traditional approach, a detection limit of 25 microM and good stability. The "Bite-and-Switch" approach combined with molecular imprinting can be used for the design of assays and sensors, selective for amino containing substances.

Biosensing Techniques↗

PubMed2001

2,4-Toluene diamines--their carcinogenicity, biodegradation, analytical techniques and an approach towards development of biosensors.

2,4-Toluene diamine (TDA), a class A carcinogen, is a major raw material for the production of toluene diisocyanate (TDI), which is one of the precursors for the production of polyurethane foams (PU). This review deals with 2,4-toluene diamine's (TDA) carcinogenicity, analytical techniques, biodegradation and use as a biosensor for biogenic and synthetic amines, emphasizing various carcinogenicity studies by 2,4-TDA on animals and humans. This review reports some publications of the analysis of body fluid samples of workers from a PU producing factory for presence of TDA and TDI, since TDI gets absorbed into the worker's body, getting metabolized into TDA. Biodegradations of 2,4-TDA by various researchers are reported and also our own research experience with biodegradation of 2,4-TDA using Aspergillus nidulans isolated from soil site at a polyurethane foam dumping site have been discussed in this review. Biosensors for various biogenic and synthetic amines are discussed.

Animals↗

PubMed2001

Cyclic voltammetric measurements of growth of Aspergillus terreus.

A detailed study using the cyclic voltammogram was done on the live cells of Aspergillus terreus. The peak current values were obtained for different days of growth and plotted against time. The response of cyclic voltammogram showed the phases of the growth of the fungus. The growth curve obtained matched well with the conventional methodology, which assesses the increase of dry weight of the organisms against time. The electrochemical method is more advantageous because it is easy to assess and consumes less time. Further the electrochemical method clearly shows the decline phase which is generally not very defined in the conventional method of assessment of the growth curve. It was confirmed by further experiments that the metabolites were responsible for the anodic peak and not the biomass. Further work is in progress in order to analyze the metabolite(s) that is/are responsible for the anodic peak.

Aspergillus↗

PubMed2000

Assessment of growth of Fusarium solani by cyclic voltammetry and possible bioanalytical applications.

Fusarium solani, the fungus isolated from polyurethane waste scraps, was studied for its voltammetric response. The peak current values were obtained for different days and were plotted against time. This electrochemical method based on the voltammetric response showed all the four phases of the growth of the fungus. The growth curve obtained matched well with the conventional methodology, which is obtained by assessing the increase of dry weight of the organisms against time. The electrochemical method is more advantageous than the conventional method because the conventional method is very time consuming and difficult to work with. Further, the electrochemical method clearly shows the decline phase, which is generally not very well defined in the conventional method of assessment of the growth curve. It was confirmed by further experiments that the metabolites were responsible for the anodic peak and not the fungal biomass. Identification of the metabolites that are responsible for the anodic peak is presently being studied.

Biodegradation, Environmental↗

PubMed1999

Isolation from genomic DNA of sequences binding specific regulatory proteins by the acceleration of protein electrophoretic mobility upon DNA binding.

We report an efficient and flexible in vitro method for the isolation of genomic DNA sequences that are the binding targets of a given DNA binding protein. This method takes advantage of the fact that binding of a protein to a DNA molecule generally increases the rate of migration of the protein in nondenaturing gel electrophoresis. By the use of a radioactively labeled DNA-binding protein and nonradioactive DNA coupled with PCR amplification from gel slices, we show that specific binding sites can be isolated from Escherichia coli genomic DNA. We have applied this method to isolate a binding site for FadR, a global regulator of fatty acid metabolism in E. coli. We have also isolated a second binding site for BirA, the biotin operon repressor/biotin ligase, from the E. coli genome that has a very low binding efficiency compared with the bio operator region.

Bacterial Proteins↗

PubMed1998

FadR, transcriptional co-ordination of metabolic expediency.

FadR is an Escherichia coli transcriptional regulator that optimizes fatty acid metabolism in response to exogenously added fatty acids. Many bacteria grow well on long-chain fatty acids as sole carbon source, but at the expense of consuming a useful structural material. Exogenous fatty acids are readily incorporated into membrane phospholipids in place of the acyl chains synthesized by the organism, and phospholipids composed of any of a large variety of exogenously derived acyl chains make biologically functional membranes. It would be wasteful for bacteria to degrade fatty acids to acetyl-CoA and then use this acetyl-CoA to synthesize the same (or functionally equivalent) fatty acids for phospholipid synthesis. This line of reasoning suggests that bacteria might shut down endogenous fatty acid synthesis on the addition of long-chain fatty acids to the growth medium. Moreover, this shutdown could be closely coupled to fatty acid degradation, such that a bacterial cell would use a portion of the exogenous fatty acid for phospholipid synthesis while degrading the remainder to acetyl-CoA. To a degree, the bacterium could both have its cake (the acyl chains for phospholipid synthesis) and eat it (to form acetyl-CoA). This scenario turns out to be true in E. coli. The key player in this regulatory gambit is FadR, a transcription factor that acts both as a repressor of the fatty acid degradation and as an activator of fatty acid biosynthesis.

Acetyl Coenzyme A↗

PubMed1998

Overproduction of a functional fatty acid biosynthetic enzyme blocks fatty acid synthesis in Escherichia coli.

beta-Ketoacyl-acyl carrier protein (ACP) synthetase II (KAS II) is one of three Escherichia coli isozymes that catalyze the elongation of growing fatty acid chains by condensation of acyl-ACP with malonyl-ACP. Overexpression of this enzyme has been found to be extremely toxic to E. coli, much more so than overproduction of either of the other KAS isozymes, KAS I or KAS III. The immediate effect of KAS II overproduction is the cessation of phospholipid synthesis, and this inhibition is specifically due to the blockage of fatty acid synthesis. To determine the cause of this inhibition, we examined the intracellular pools of ACP, coenzyme A (CoA), and their acyl thioesters. Although no significant changes were detected in the acyl-ACP pools, the CoA pools were dramatically altered by KAS II overproduction. Malonyl-CoA increased to about 40% of the total cellular CoA pool upon KAS II overproduction from a steady-state level of around 0.5% in the absence of KAS II overproduction. This finding indicated that the conversion of malonyl-CoA to fatty acids had been blocked and could be explained if either the conversion of malonyl-CoA to malonyl-ACP and/or the elongation reactions of fatty acid synthesis had been blocked. Overproduction of malonyl-CoA:ACP transacylase, the enzyme catalyzing the conversion of malonyl-CoA to malonyl-ACP, partially relieved the toxicity of KAS II overproduction, consistent with a model in which high levels of KAS II blocks access of the other KAS isozymes to malonyl-CoA:ACP transacylase.

3-Oxoacyl-(Acyl-Carrier-Protein) Synthase↗

PubMed1997

A comparison of peak expiratory flow rates obtained with the Morgan Spirocheck and a Mini Peak Flow Meter.

To examine whether the normal Indian standards of Peak Expiratory Flow Rate (PEF) established with the peak flow meter were applicable to PEF values obtained with the Morgan Spirocheck, a comparison was made of PEF(S) measured with the Spirocheck with PEF (M) obtained with a Mini Peak Flow Meter. Data were obtained in 92 subjects (53 males between 20 and 84 years of age and 39 females between 21 and 75 years), comprising staff of the institute, patients undergoing treatment for chronic arthritis, and the accompanying relatives. There was a highly significant correlation between the two values. The values were identical in 14 subjects; the differences between the values were within 10% in 54 subjects, and in excess of 10% in only 20 subjects. In the whole group, the mean and standard error were 363.5 +/- 14.7 and 384 +/- 15.2 lit/min for PEF (S) and PEF (M) respectively. The regression equation PEF (M) = 0.961 x PEF (S) + 34.9 will enable estimation of expected value of PEF (M) corresponding to the observed value of PEF (S) when a standard obtained with a Peak Flow Meter in used assess abnormality in a patient.

Adult↗

PubMed1985