Clinical utility of the interaction between lectin and serum prostate specific antigen in prostate cancer.

The present investigation was a lectin-based diagnosis of malignant prostate cancer (PC) by the interaction of phytohemagglutinin (PHA lectin) from Phaseolus vulgaris with the glycan part of serum prostate specific antigen (PSA) of patients with prostatic disorder. This was confirmed by the interaction between PHA and purified PSA obtained from serum by electrophoretic separation and finally by HPLC chromatography. The precipitate of carbohydrate content after binding of PHA with purified PSA of PC was significantly higher than that of benign prostate hyperplasia (BPH) and/or normal serum PSA. The results suggest that there may be a striking difference in glycosylation pattern of PSA between BPH and PC. The cut off value > or = 10 microg/ml of the carbohydrate content of PHA-PSA precipitate indicates strong suspicion for PC irrespective of total serum PSA cut off level > or = 4.0 ng/ml by conventional immunoassay method and this may be taken as a guideline in differentiating PC and BPH. Key words: prostate cancer, BPH, PSA, lectin.

Osmotic adjustment increases water uptake, remobilization of assimilates and maintains photosynthesis in chickpea under drought.

Eight chickpea advanced breeding lines (ABLs) and their parents were evaluated for osmotic adjustment (OA), leaf carbohydrates and gas exchange under dryland field . These (ABLs) were derived from crosses between CTS 60543 x Kaniva and Tyson x Kaniva. Mean leaf water potential (LWP) fell down from -1.00 MPa at pre-stress level to about -2.25 MPa during terminal stress. Relative water content (RWC) showed periodic changes with alternate decrease or increase at certain interval, which also influenced the values of OA (low or high) in number of genotypes e.g. Kaniva, CTS 60543, Tyson and M 75. Significant variation in OA ranging 0.45 to 0.88 MPa was observed at high level of stress at -2.5 MPa. However, none of the genotypes showed stability of OA over the period of stress. Leaf starch declined even at mild stress (LWP, -1.6 MPa) resulting in an increase in hexose sugars and activation state of sucrose-phosphate synthase (SPS) that led to accumulation of sucrose. Both photosynthesis (Pmax) and transpiration decreased concurrently in two chickpea lines M 129 and Tyson with increasing water stress. However, rate of decline in the photosynthesis slowed down even drought was further intensified. The observed periodic changes in OA, RWC and photosynthesis appeared to be associated with drought-induced changes in SPS and carbohydrates which modify water uptake of the leaves.

Production and metabolism of indole acetic acid in roots and root nodules of Phaseolus mungo.

The mature root nodules of Phaseolus mungo (L.), a leguminous pulse, contain higher amount of indole acetic acid (IAA) than non-nodulated roots. The tryptophan pool present in the mature nodule and young roots might serve as a precursor for the IAA production. Presence of IAA metabolising enzymes - IAA oxidase and peroxidase - indicate the metabolism of IAA in the nodules and roots. In culture, the symbiont, isolated from the nodules, produced a high amount of IAA, when tryptophan was supplied in the medium as a precursor. The symbiont preferred l-isomer over the dl- or d-isomer of tryptophan for IAA production. The important physiological implication of the IAA production in the legume-Rhizobium symbiosis is discussed.

Enhancement of lectin-erythrocyte agglutination by gums.

The erythroagglutinating activity of purified Vicia faba lectin was enhanced in the presence of gums; gum guar caused the highest enhancement. Circular dichroism probe demonstrated 40-57% beta-conformation and 4-23% alpha-conformation of the lectin at pH 7.2 depending upon the analytical methods used. The beta-conformations of untreated and modified V. faba lectins were increased in the presence of gums. The mixing of gum guar with lectin and with modified lectin, respectively, led to the highest values of beta-conformational change in the protein molecule, thereby increasing the number of receptor sites of the lectin molecule. The enhancement of the activity of V. faba lectin in the presence of gum guar might be due to the conformational change of the protein molecule.

Purification and partial characterization of an erythroagglutinin from the hemolymph of scorpion, Heterometrus bengalensis.

An erythroagglutinin from the hemolymph of the scorpion, Heterometrus bengalensis, has been purified by gel filtration and ion-exchange chromatography. Its homogeneity has been demonstrated by polyacrylamide gel electrophoresis. The purified agglutinin appears to be a monomeric protein having a possible molecular weight between 146,000 and 148,000. It has no divalent cation requirement for erythroagglutination. The erythroagglutination is not inhibited by saccharides, glycoproteins and mucin. Identical erythroagglutination pattern is obtained with normal as well as neuraminidase treated erythrocytes.

Characterization of a naturally occurring protease inhibitor in the hemolymph of the scorpion, Heterometrus bengalensis.

A protease inhibitor has been purified by ultracentrifugation, affinity chromatography on trypsin-sepharose 4B, and chromatofocusing on PBE-94 from hemolymph of the scorpion Heterometrus bengalensis. Homogeneity of the protease inhibitor was demonstrated by high performance liquid chromatography (HPLC). The protease inhibitor is a monomeric glycoprotein with a molecular weight of 120,000 dalton, which is stable between pH 4 and pH 8. The molecule inhibits serine proteases like trypsin and alpha-chymotrypsin and shows a noncompetitive mode of inhibition towards trypsin, with a Ki value of 6.1 x 10(-6) mM. Amino acid analysis shows a preponderance of aspartic acid, glutamic acid, serine, and glycine. The protease inhibitor is efficient in inhibiting phenoloxidase activity in both the hemolymph and the isolated phenoloxidase. Melanin synthesis by phenoloxidase may be influenced by this protease inhibitor.

Acetylcholinesterase (EC 3.1.1.7), a neurotransmitter enzyme in scorpion hemolymph.

Acetylcholinesterase (AchE: EC 3.1.1.7) was identified and purified from the hemolymph of the scorpion Heterometrus bengalensis. The purity of the enzyme was determined by polyacrylamide gel electrophoresis (PAGE). The molecular weight of the enzyme, determined by sodium dodecyl sulfate-PAGE, was 80,000. The purified AchE hydrolysed acetylthiocholine iodide, but it did not react with butyrylthiocholine iodide. BW284C51, a specific inhibitor of AchE, strongly inhibited the enzyme. The known inhibitor (tetramonoisopropylpyrophosphortetramide) of pseudocholinesterase did not produce any inhibition of the enzyme activity. The purified AchE of scorpion hemolymph was vulnerable to high substrate concentration. The presence of Cu2+ and Ni2+ reduced the enzyme activity, whereas the metal ion, Sn2+, enhanced AchE activity. Ca2+ produced neither inhibition nor activation. (Na+, K+)-ATPase and Mg2+-ATPase activities were greatly enhanced by the purified AchE.

Possible mechanism for the inhibition of lectin-erythrocyte interaction in presence of endogenous lectin receptor.

The presence of hydrophobic sites in the lectin-I molecule was indicated by hydrophobic probes like 1-anilinonapthalene-8-sulfonic acid (ANS), 2-p-toluidinyl napthalene-6-sulfonic acid (TNS). N-phenyl-1-napthylamine (NA) and rose bengal (RB). This was further confirmed by amino acid modifications in the hydrophobic region of the lectin-I molecule. The binding of ANS, TNS, NA and RB to lectin-I was affected in the presence of NaCl. The involvement of hydrophobic interactions in rice-bean lectin-I-endogenous lectin receptor (ELR) complex were indicated by alterations in the circular dichroism and fluorescence emission spectra. The percentage of beta-conformation (55-63%) of lectin-I was decreased by addition of ELR. ELR on reacting with lectin-I reduced the fluorescence emissions of the hydrophobic probes while fluorescence emission of ANS, TNS, NA and RB were greatly enhanced in presence of lectin-I alone. N-aceyl-galactosamine did not change the fluorescence emissions of any of the hydrophobic probes in presence or in absence of lectin-I. This demonstrates that carbohydrate and hydrophobic sites may be different and non-interacting. It is proposed that the ELR in reacting with lectin-I, induced conformational changes in the lectin-I molecule and thereby affected its erythroagglutinating activity with human blood group "A" erythrocytes.

Inhibition of rat liver mitochondrial monoamine oxidase by chloramphenicol and 2-amino-1-p-nitrophenylpropane-1,3-diol.

Inhibition of rat liver mitochondrial monoamine oxidase by chloramphenicol and its hydrolytic product, 2-amino-1-p-nitrophenylpropane-1,3-diol, was studied. The enzyme activity and its inhibition by these two compounds were optimum at pH 7.0 after preincubation for 60 min, the time needed for maximum enzyme--inhibitor complex formation. Enzyme activity could be restored after prolonged dialysis. Monamine oxidase inhibition by chloramphenicol and its hydrolytic product was noncompetitive and reversible. Deamination of various monamines was not to the same degree by these compounds. Of the different antimicrobials studied, only chloramphenicol and its hydrolytic produced has a strong inhibitory effect on monoamine oxidase.

Spondylolisthesis in osteogenesis imperfecta due to pedicle elongation: report of two cases.

STUDY DESIGN: Two cases of spondylolisthesis due to severe elongation of pedicles in osteogenesis imperfecta are reported. OBJECTIVE: To describe an unusual type of spondylolisthesis and its successful management. SUMMARY OF BACKGROUND DATA: Spondylolisthesis is known to occur in osteogenesis imperfecta. Reports in the literature are few. Pedicle elongation resulting in spondylolisthesis has only been reported once. There is no report of a successful treatment of this type of spondylolisthesis resulting from pedicle elongation. METHOD: Two adolescent girls suffering from osteogenesis imperfecta presented with lower back pain and thoracic scoliosis. One of them had high-grade spondylolisthesis of L3-L4, L4-L5, and L5-S1, with a thoracic scoliosis. She was treated with anterior interbody fusion L3 to sacrum without instrumentation. Later, progression of her scoliosis required combined anterior and posterior instrumented fusion T1 to L1. The other girl presented with L5 spondylolisthesis and thoracic scoliosis. She has back pain as well as neurologic symptoms in her legs and is waiting for surgery. RESULTS: At the 3-year follow-up the operated patient is symptom-free from her spine and there has been no progression of either deformity. CONCLUSION: High-grade spondylolisthesis due to elongation of pedicle in osteogenesis imperfecta is uncommon. Anterior interbody fusion of all the involved motion segments has produced good pain relief and arrested deformity progression at the 3-year follow-up.

Indole acetic acid production by a Rhizobium species from root nodules of a leguminous shrub, Cajanus cajan.

A Rhizobium sp. isolated from the root nodules of a leguminous pulse-yielding shrub, Cajanus cajan, was found to produce high amounts (99.7 microg/ml) of indole acetic acid (IAA) during growth in basal medium supplemented with L-tryptophan. The Rhizobium sp. was a fast growing species which reached its stationary phase at 28 h. The IAA production could be increased upto 653.3% over control by supplementing the carbon-free incubation medium with glucose (5 g/l), NiCl2 (10 microg/ml) and glutamic acid (0.5 g/l). The possible role of rhizobial IAA on the rhizobia-legume symbiosis is discussed.

Indoleacetic acid production by Rhizobium sp. from a leguminous tree, Erythrina indica.

Rhizobium sp. isolated from the root nodules of Erythrina indica Lamk., a leguminous tree, produced large amounts of indoleacetic acid (IAA) from tryptophan. L-Tryptophan (2 mg/ mL) was preferred to DL-tryptophan for IAA synthesis. Attempts were made to optimize the cultural requirements for an accelerated IAA production. An enrichment of the carbon-free incubation medium with maltose (1 %), MnSO4 x H2O (5 microg/mL), and Triton X-100 (100 ppm) promoted the synthesized IAA content of the medium by 480 %.

Extracellular polysaccharide production by Azorhizobium caulinodans from stem nodules of leguminous emergent hydrophyte Aeschynomene aspera.

The Azorhizobium caulinodans isolated from the stem nodules of a leguminous emergent hydrophyte, Aeschynomene aspera, produced a large amount of extracellular polysaccharides (EPS) in yeast extract basal medium. Maximum EPS production was at the stationary phase of growth. EPS production was increased by 919% over control when the medium was supplemented with sucrose (1.5%), D-biotin (1 microgram/ml) and casamino acid (0.1%). EPS contained rhamnose and arabinose. Possible role of the azorhizobial EPS production in the stem nodule symbiosis is discussed.

Growth behaviour and bioproduction of indole acetic acid by a Rhizobium sp. isolated from root nodules of a leguminous tree Dalbergia lanceolaria.

The Rhizobium sp. isolated from healthy and mature root nodules of a leguminous tree, Dalbergia lanceolaria Linn. f., preferred mannitol and KNO3 for growth as carbon and nitrogen sources, respectively. The bacterium produced a high amount (22.3 microg/ml) of indole acetic acid (IAA) from L-tryptophan supplemented basal medium. Growth and IAA production started simultaneously. IAA production was maximum at 20 hr when the bacteria reached the stationary phase of growth. Cultural requirements were optimized for maximum growth and IAA production. The IAA production by the Rhizobium sp. was increased by 270.8% over control when the medium was supplemented with mannitol (1%,w/v), SDS (1 microg/ml), L-asparagine (0.02%,w/v) and biotin (1 microg/ml) in addition to L-tryptophan (2.5 mg/ml). The possible role of IAA production in the symbiosis is discussed.

Parasympatholytic activity of psychoactive drugs in rat brain mitochondria.

Chlorpromazine, imipramine and amphetamine at a concentration of 0.66, 1.33 and 13.3 x 10(4) M in vitro inhibited acetyl cholinesterase activity by 16, 23 and 31% respectively in rat brain mitochondria. No change in enzyme activity was induced by these drugs in vivo. There is little cholinergic facilitation through acetylcholinesterase inhibition in the presence of psychoactive drugs.

R-factor in Proteus vulgaris from ulcerative disease of fish, Channa punctatus.

A Proteus vulgaris isolated from external ulcers of the fresh water fish Channa punctatus showed multidrug resistance and heavy metal tolerance. The isolate from the ulcer showed resistance to chloramphenicol (Ch), nalidixic acid (Nx), streptomycin (Str) and tetracycline (Tet) with minimum inhibitory concentration (MIC) values of 750, 150, 75 and 125 microg/ml, respectively. The isolate showed growth in medium containing cadmium (Cd2+), up to a concentration of 2.5 mM indicating its heavy metal tolerance. Resistance to Ch, Str, Tet and Cd2+ of the isolate was lost after plasmid curing. Presence of plasmid DNA in the wild type and its absence in the cured P. vulgaris suggested that the resistance were plasmid mediated.

Serine proteinase from rice bean.

A trypsin like serine-proteinase of M(r) 16,000 Da, optimally active at pH 8.4 on N-benzoyl-arginine ethyl ester (BAEE) was purified from 4-day old germinated seeds of rice bean, Vigna umbellata (Thunb), by ammonium sulphate precipitation, gel filtration, ion-exchange chromatography and by high performance liquid chromatography (HPLC). The purity of the enzyme was checked by polyacrylamide gel electrophoresis (PAGE). The enzyme activity was studied on natural substrates like casein, haemoglobin and vicilin, a rice bean storage protein. The activity of the enzyme was completely inhibited by phenylmethylsulfonyl fluoride, but not by iodoacetamide and HgCl2, suggesting it to be a serine protease. Loss of activity in presence of EDTA was reversed by addition of Ca2+.

Separation and identification of thyroid autoantibodies in patients with thyroid disorders by hydrophobic column.

A method has been developed to separate and identify thyroglobulin autoantibody (TgAb) and thyroid peroxidase autoantibody (TPOAb) in serum obtained from normal and autoimmune thyroid diseases using phenyl Sepharose CL-4B hydrophobic column. The protein peaks obtained from hydrophobic column were identified as TgAb and TPOAb by comparing the elution profile of commercially purified standard thyroid autoantibodies. The similarity of the inhibitory effects of eluted proteins and of standard thyroid autoantibodies on lectin concanavalin A-RBC interaction confirmed the separation of TPO-Ab and TgAb by the hydrophobic, column. The eluted fractions from the hydrophobic column were estimated by the radio immunoassay (RIA) to confirm the presence of both auto-antibodies. This hydrophobic column method offers an advantage of visual inspection of this autoantibodies by graphic representation of peak height along with their estimation in autoimmune thyroid disorders.

Phenotyping chickpeas and pigeonpeas for adaptation to drought.

The chickpea and pigeonpea are protein-rich grain legumes used for human consumption in many countries. Grain yield of these crops is low to moderate in the semi-arid tropics with large variation due to high GxE interaction. In the Indian subcontinent chickpea is grown in the post-rainy winter season on receding soil moisture, and in other countries during the cool and dry post winter or spring seasons. The pigeonpea is sown during rainy season which flowers and matures in post-rainy season. The rainy months are hot and humid with diurnal temperature varying between 25 and 35°C (maximum) and 20 and 25°C (minimum) with an erratic rainfall. The available soil water during post-rainy season is about 200-250 mm which is bare minimum to meet the normal evapotranspiration. Thus occurrence of drought is frequent and at varying degrees. To enhance productivity of these crops cultivars tolerant to drought need to be developed. ICRISAT conserves a large number of accessions of chickpea (>20,000) and pigeonpea (>15,000). However only a small proportion (<1%) has been used in crop improvement programs mainly due to non-availability of reliable information on traits of economic importance. To overcome this, core and mini core collections (10% of core, 1% of entire collection) have been developed. Using the mini core approach, trait-specific donor lines were identified for agronomic, quality, and stress related traits in both crops. Composite collections were developed both in chickpea (3000 accessions) and pigeonpea (1000 accessions), genotyped using SSR markers and genotype based reference sets of 300 accessions selected for each crop. Screening methods for different drought-tolerant traits such as early maturity (drought escape), large and deep root system, high water-use efficiency, smaller leaflets, reduced canopy temperature, carbon isotope discrimination, high leaf chlorophyll content (drought avoidance), and breeding strategies for improving drought tolerance have been discussed.