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Biomedical subjects

S Basu

Publications and source records attributed to S Basu.

At least 379 records · Page 21Linked to original sources
PubMed1989

Stimulation of DNA chain initiation by a protein factor (NPF-1) from rat liver of different ages.

DNA Polymerase-alpha/primase complexes have been isolated from human neuroblastoma IMR-32, embryonic chicken brains (ECB) and rat prostate tumor PA-3 cells. In the presence of (NH4)2SO4 the major part (90%) of primase activity is released from the Pol-alpha/primase complex. A novel hydrophobic interaction column was used for purification of the primase from PA-3 cells. A nuclear protein factor (NPF-1) that stimulates DNA pol-alpha/primase activity has been purified from rat liver of various ages (3-6 months). The nuclear protein factor which only stimulates the primase activity is under investigation. The monoclonal antibodies (SJK 132-20 and 237-71) were used to detect DNA pol-alpha polypeptides from 11- to 19-day-old embryonic chicken brains.

Aging↗
PubMed1989

Preliminary studies on excretory components of a virulent strain (F-2) of Pseudomonas solanacearum.

The excretory material (EM) was isolated from the culture medium of a virulent strain (F-2) of Pseudomonas solanacearum. Electrophoretic studies depicted the heterogeneous nature of EM and the presence of released lipopolysaccharide in it. Both the exopolysaccharide and the released LPS contained D-galactose as the major sugar constituent together with D-glucose, N-acetyl-D-galactosamine and N-acetyl-D-glucosamine. In addition, L-rhamnose was present as a constituent sugar of the released LPS.

Bacterial Proteins↗
PubMed1988

Pyridoxal 5'-phosphate mediated inactivation of Escherichia coli DNA polymerase I: identification of lysine-635 as an essential residue for the processive mode of DNA synthesis.

Inactivation of Escherichia coli DNA polymerase I by pyridoxal 5'-phosphate treatment results from its reactivity at multiple lysine residues. One of these residues, lysine-758, has been shown to be located at the substrate binding site in DNA polymerase I [Basu, A., & Modak, M. J. (1987) Biochemistry 26, 1704-1709]. We now demonstrate that lysine-635 is another important target of pyridoxylation; modification of this site results in decreased rates of DNA synthesis. Addition of template-primer with or without substrate deoxynucleoside triphosphate protects lysine-635 from pyridoxylation. Analysis of the initiation versus elongation phase of DNA synthesis by lysine-635-modified enzyme revealed that elongation of the DNA chain is severely affected by the lysine-635 modification. We therefore conclude that this lysine residue plays an important role in the processive mode of DNA synthesis by E. coli DNA polymerase I.

Binding Sites↗
PubMed1988

Chemical-modification studies of a unique sialic acid-binding lectin from the snail Achatina fulica. Involvement of tryptophan and histidine residues in biological activity.

A unique sialic acid-binding lectin, achatininH (ATNH) was purified in single step from the haemolymph of the snail Achatina fulica by affinity chromatography on sheep submaxillary-gland mucin coupled to Sepharose 4B. The homogeneity was checked by alkaline gel electrophoresis, immunodiffusion and immunoelectrophoresis. Amino acid analysis showed that the lectin has a fairly high content of acidic amino acid residues (22% of the total). About 1.3% of the residues are half-cystine. The glycoprotein contains 21% carbohydrate. The unusually high content of xylose (6%) and fucose (2.7%) in this snail lectin is quite interesting. The protein was subjected to various chemical modifications in order to detect the amino acid residues and carbohydrate residues present in its binding sites. Modification of tyrosine and arginine residues did not affect the binding activity of ATNH; however, modification of tryptophan and histidine residues led to a complete loss of its biological activity. A marked decrease in the fluorescence emission was found as the tryptophan residues of ATNH were modified. The c.d. data showed the presence of an identical type of conformation in the native and modified agglutinin. The modification of lysine and carboxy residues partially diminished the biological activity. The activity was completely lost after a beta-elimination reaction, indicating that the sugars are O-glycosidically linked to the glycoprotein's protein moiety. This result confirms that the carbohydrate moiety also plays an important role in the agglutination property of this lectin.

Amino Acids↗
PubMed1988

DNA binding domain of Escherichia coli DNA polymerase I: identification of arginine-841 as an essential residue.

To identify the DNA binding site(s) in Escherichia coli DNA polymerase I (pol I) (Klenow fragment), we have used an active-site-directed reagent, phenylglyoxal (PG), which specifically reacts with arginine residues. Preincubation of DNA pol I with PG resulted in the loss of polymerase, 3'-5'-exonuclease, and DNA binding functions. Furthermore, the presence of DNA but not deoxynucleoside triphosphates protected the enzyme from inactivation. Labeling studies with [7-14C]PG indicated that two arginine residues were modified per mole of enzyme. In order to locate the site of PG modification, we digested the PG-treated enzyme with trypsin and V-8 protease. The resulting peptides from each digest were then resolved on reverse-phase hydrophobic columns. An appearance of a new peptide peak was observed in both tryptic and V-8 protease digests. Since inclusion of template-primer during PG modification of enzyme blocks the appearance of these peaks, these peptides were concluded to represent the template-primer binding domain of pol I. Indeed, the extent of inactivation of enzyme by PG treatment correlated very well with the quantitative increase in the new tryptic peptide peak. Amino acid composition analysis of both tryptic peptide and V-8 peptide revealed that the two peptides were derived from the same general region; tryptic peptide spanned between residues 837 and 857 while V-8 peptide spanned between residues 841 and 870 in the primary sequence of pol I. Sequence analysis of tryptic peptide further identified arginine-841 as the site of PG modification, which implicates this residue in the DNA binding function of pol I.

Arginine↗
PubMed1988

Effect of blood bonding on bursting strength of laser-assisted microvascular anastomoses.

This experiment evaluated the influence of blood on the weld strength of laser-assisted microvascular anastomoses (LAMA). Rat femoral arteries were anastomosed end-to-end by either direct laser welding (group I) or by a blood-bonded technique (group II) whereby fresh blood was applied to the vessel edges before laser exposure. Bursting strength was measured at 0, 1, and 24 hours and at 3 and 7 days by infusing methylene blue into the vessel while pressure was monitored. The results showed significantly increased bursting strength in group II compared with group I at 0 hour and 7 days (P less than 0.05). There was a significant increase in bursting strength in group I from 0 hour to 1 hour (P less than 0.05). It is concluded that blood-bonding enhances the early bursting strength of LAMAs and may facilitate arterial wall healing.

Anastomosis, Surgical↗
PubMed1988

Enhancement of CO2 laser microvascular anastomoses by fibrin glue.

Laser-assisted microvascular anastomoses (LAMA) are characterized by low early bursting strength and high aneurysm rates. The effects of fibrin glue on bursting strength (BS), patency, and aneurysm rate of LAMAs were compared to standard suture and laser anastomosis. Rat femoral arteries (0.9-1.1 mm) were anastomosed end-to-end by three methods: (1) conventionally with 8 to 10 interrupted 10-O nylon sutures; (2) 3 stay sutures and CO2 laser (spot size, 0.275 mm; pulse, 0.2 sec, 80 mW); and (3) cryoprecipitated fibrinogen, 35 mg/cc, crystallized thrombin, CaCl, 20 mg/cc, aprotinin (2000 kIU/cc) applied to weld site in conjunction with laser weld as in (2) above. Patency, aneurysms, and histology were evaluated at 3 weeks, and BS (mm Hg) was measured in six additional vessels at 1 and 24 hr. There was no statistically significant difference in patency rates. Both the suture and fibrin glue groups had significantly higher 1 and 24 hr bursting strengths (P less than 0.05) and significantly lower aneurysm rates (P less than 0.001) than standard laser. There was no significant difference in bursting strength between suture and fibrin glue groups. Histology in the fibrin glue group showed medial damage similar to the LAMA and calcification of aneurysmal vessels. Fibrin glue enhancement of LAMAs produces equal patency, higher early bursting strength, and fewer aneurysms at 3 weeks compared to conventional laser.

Anastomosis, Surgical↗
PubMed1988

Solubilized glycosyltransferases and biosynthesis in vitro of glycolipids.

The assembly of most of the ceramide-linked glycolipids (GSLs) in eukaryotic cells occurs in Golgi bodies. At least 18 different glycolipid:glycosyltransferases (GSL:GLTs) have been characterized, 10 of which have been solubilized. These GLTs can be classified into 2 distinct groups: 1) GLTs dedicated to either Dol-P-P-sugar(s) or ceramide-linked sugar(s); and 2) GLTs with dual loyalties (i.e., they compete with glycolipid- and glycoprotein-bound oligosaccharides). Studies with solubilized and purified GalNAcT-1 and GalNAcT-2 from embryonic chicken brains prove that GalNAcT-1 (UDP-GalNAc:GM3 beta 1-4GalNAcT) is specific for GSL, whereas GalNAcT-2 (UDP-GalNAc:Gb3 beta 1-3GalNAcT) can transfer to an oligosaccharide containing the alpha-linked terminal galactose. Similarly, GalT-3 (UDP-Gal:GM2 beta 1-3GalT) is more specific for ganglio-oligosaccharide and GalT-4 (UDP-Gal:Lc3 beta 1-4GalT) can transfer galactose to N-acetylglucosamine linked to p-nitrophenol, glycolipid or glycoprotein. Both GalT-3 and GalT-4 have been separated and purified from embryonic chicken brains. Studies with solubilized SAT-4 and SAT-3, from bovine spleen and embryonic chicken brains, respectively, suggest the existence of 2 different gene-expressed alpha 2-3SATs. The newly discovered FucT-3 (GDP-Fuc:NeuGc-iLc6-alpha 1-3FucT) from human colon carcinoma (Colo-205) has also been solubilized and separated from other GSL:GLTs. Using a new activity gel-Western blot combined technique, the molecular mass of this FucT-3 was determined to be 105 kDa.

Animals↗
PubMed1988

Modification of prostaglandin F-2 alpha synthesis and release in the ewe during the initial establishment of pregnancy.

Pregnant (N = 10) and non-pregnant (N = 10) ewes were bled every 2 h from Days 12 to 17 after oestrus (oestrus = Day 0). Plasma concentrations of progesterone, 15-keto-13,14-dihydro-PGF-2 alpha and 11-ketotetranor-PGF metabolites were determined in all samples. The number of PGF-2 alpha pulses in non-pregnant ewes was 8.2 +/- 0.4 (mean +/- s.e.m.) with an interpulse interval of 10.7 +/- 0.7 h. Two or 3 pulses of low frequency (interpulse interval = 13.4 +/- 1.6 h) occurred in most non-pregnant ewes before the onset of luteolysis; the interpulse interval then decreased to 7.9 +/- 0.4 h for the 6.0 +/- 0.3 pulses temporally associated with luteolysis. In contrast, the number of PGF-2 alpha pulses in pregnant ewes was lower (2.5 +/- 0.7, 0-8) and the interpulse intervals longer (18.9 +/- 6.1 h). Most pulses occurred on Days 14 and 15 in the pregnant and non-pregnant ewes. The mean concentrations of both PGF-2 alpha metabolites in non-pregnant ewes were highest on Day 15 while basal levels of both metabolites remained constant at all times. In pregnant ewes, the mean concentrations of both metabolites were highest on Day 14; basal concentrations of both metabolites were also highest on Day 14. The mean concentrations of 15-keto-13,14-dihydro-PGF-2 alpha were higher in pregnant than in non-pregnant ewes on Days 13 and 14 (P less than 0.05) and higher in non-pregnant than pregnant ewes on Day 15 (P less than 0.05). The basal concentrations of the 15-keto metabolite were higher in pregnant than non-pregnant ewes at Days 13, 14, 15, 16 and 17 (P less than 0.05). Both the mean and the basal concentrations of 11-ketotetranor-PGF metabolites were higher in pregnant than in non-pregnant ewes on Day 14 (P less than 0.05). It is concluded that uterine production of PGF-2 alpha peaks at Days 14-15 after oestrus in pregnant and non-pregnant ewes. Patterns of release differ, however, in that non-pregnant ewes have a pulsatile PGF-2 alpha pattern superimposed on a constant baseline, while pregnant ewes have an increasing basal secretory pattern which is more nearly continuous, i.e. not pulsatile in form. Modification of pulsatile PGF-2 alpha synthesis and release is therefore a key aspect of prolongation of luteal function at the beginning of pregnancy in the ewe.

Animals↗
PubMed1988

In vitro bursting strength studies of laser-welded tissue and comparison with conventional anastomosis.

We compared the mechanical integrity of microvascular anastomoses created with a carbon dioxide (CO2) laser with conventional suture anastomoses. Seventy rat femoral artery segments (35 lased and 35 sutured) were harvested at 1, 24, and 72 hours, and 1, 3, 6, and 12 weeks postoperatively. These segments were subjected to increasingly higher in vitro intraluminal hydrostatic pressures (bursting pressure). Conventionally sutured anastomoses exhibited significantly increased ability (p less than 0.05) to withstand greater bursting pressures than the laser-welded tissue up to 3 weeks postoperatively. After the third postoperative week, the laser anastomoses demonstrated increased bursting pressures compared with the conventional anastomoses. At the end of the 12-week period both groups demonstrated an ability to withstand supraphysiologic pressures in excess of 2000 mm Hg.

Anastomosis, Surgical↗
PubMed1988

Somatosensory evoked potentials and spinal cord perfusion pressure are significant predictors of postoperative neurologic dysfunction.

Paraplegia after thoracoabdominal aneurysm repair can occur in 3% to 40% of patients. This study investigated the efficacy of cerebrospinal fluid (CSF) drainage to protect the spinal cord during aortic cross-clamping (AXC) and the interrelationship between drainage, spinal cord perfusion pressure (SCPP), and changes in somatosensory evoked potentials (SEP) in a canine model of spinal cord ischemia. SCPP was defined as the mean distal aortic pressure minus the CSF pressure. In the experimental group, CSF was drained before AXC. SEP changes were quantitated as time to latency increase of 10% (L-10) and time to complete SEP loss. Drainage of CSF had no significant effect on the distal aortic pressure but significantly increased SCPP from 9.4 to 21.8 mm Hg and decreased the incidence of postoperative neurologic injury. Ischemic SEP changes were highly significant predictors of postoperative neurologic injury, occurring more than two times earlier in the paralyzed and paraparetic animals. Dogs without neurologic injury had significantly higher SCPP, delayed L-10 time, and delayed SEP loss.

Animals↗
PubMed1987

An unique specificity of a sialic acid binding lectin AchatininH, from the hemolymph of Achatina fulica snail.

A sialic acid-binding lectin, AchatininH, from the hemolymph of Achatina fulica snail is found to be highly specific for 9-0-acetyl sialic acid. The binding specificity of AchatininH distinguishes it from other known sialic-acid specific lectins which usually show a broader range of specificity for sialic acid. It is even better than crab lectin which shows specificity for both 4- and 9-0-acetylated derivatives of sialic acid. This limited specificity of AchatininH appear to account for the fact that it agglutinates only rabbit, rat and guinea pig erythrocytes which contain 9-0-acetylated sialic acid but not horse (mainly contain 4-0-acetylated sialic acid), human, monkey, sheep, goat and chicken erythrocytes which contain either N-acetyl or N-glycolyl neuraminic acid but no 0-acetylated derivatives. This finding was further supported by the potent inhibition of hemagglutination by free 9-0-acetylated neuraminic acid and by several glyco shingolipids of human origin having 0-acetylated sialic acid.

ABO Blood-Group System↗