Mediated uptake of folate by a high-affinity binding protein in sublines of L1210 cells adapted to nanomolar concentrations of folate

An L1210 cell line (JT-1), which can grow in medium supplemented with 1 nM folate, has been isolated. These cells exhibit a slower growth rate than folate-replete parental cells and have a lower ability to transport folate or methotrexate via the reduced folate transport system. Measurements at nanomolar concentrations of folate revealed that the adapted cells have acquired a high-affinity folate-binding protein. Binding to this component at 37 degrees C was rapid and reached a maximum value after 30 min which corresponded in amount to 0.23 +/- 0.3 pmol/mg protein, and excess unlabeled folate added 30 min subsequent to the [3H]folate led to a rapid release of the bound substrate. Radioactivity bound to or released from the cells after 30 min at 37 degrees C remained as unmetabolized folic acid. Binding was also rapid at 0 degrees C but uptake at the plateau was only one-half the value obtained at 37 degrees C. Half-maximal saturation of the binding component (KD) occurred at a folate concentration of 0.065 nM at pH 7.4, while the affinity for folate decreased 30-fold when the pH was reduced to 6.2 (KD = 2.0 nM). 5-Methyltetrahydrofolate was also bound by this component (Ki = 13 nM at pH 7.4) but with a much lower affinity than for folate, while progressively weaker interactions were observed with 5-formyltetrahydrofolate (Ki = 45 nM) and methotrexate (Ki = 325 nM). When the same adaptation procedure was performed with limiting amounts of 5-formyltetrahydrofolate, two additional cell lines, JT-2 and JT-3, were isolated which expressed elevated levels of the folate-binding protein. The binding activity of the latter cells was 0.46 and 1.4 pmol/mg protein, respectively. When the level of binding protein was compared in cells grown at different concentrations of folate, an increase in medium folate from 1 to 500 nM caused a sevenfold reduction in binding activity in the JT-3 cell line, while these same growth conditions had no effect on binding by the other cells. These results indicate that L1210 cells adapted to low concentrations of folate or 5-formyltetrahydrofolate contain elevated levels of a high-affinity binding protein and that this protein is able to mediate the intracellular accumulation of folate compounds. L1210 cells thus appear to have two potential uptake routes for folate compounds, the previously characterized anion-exchange system and a second route mediated by a high-affinity binding protein.(ABSTRACT TRUNCATED AT 400 WORDS)     

Synthesis of functional human immunodeficiency virus tat protein in baculovirus as determined by a cell-cell fusion assay.

The human immunodeficiency virus tat protein is a strong trans-activator of the expression of mRNAs originating from the viral long terminal repeat. We have expressed the first 72 amino acids (coding exon 1) of this protein in eucaryotic Spodoptera frugiperda SF9 cells by using a baculovirus vector, Autographa californica nuclear polyhedrosis virus. We show that the baculovirus vector stably produced the 72-amino-acid form of the tat protein but was unable to stably synthesize a larger 101-amino-acid full-length version of the same polypeptide. The 72-amino-acid tat protein, when introduced into mammalian fibroblasts by using a cell-cell fusion technique, functionally trans-activated the expression of the human immunodeficiency virus long terminal repeat.     

Influence of the food plants on the degree of parasitism of larvae ofHeliothis armigera byCotesia kazak

Influence of the food plants ofHeliothis armigera (Hb.) on the degree of parasitism by exotic parasiteCotesia kazak Telenga (Hymenoptera: Braconidae) was studied in cages in the laboratory on 7 food plants such as cotton (Gossypium hirsutum L.), tomato (Lycopersicon esculentum Mill), okra [Abelmoschus esculentus (L.) Moench], Dolichos (dolichos lablab L.), pigeonpea [Cajanus cajan (L.) Millsp.], Cowpea (Vigna unquiculata (L.) and chickpea (Cicer arietium L.). To determine the preference of the parasite 2 test methods were employed. In single plant choice test cotton was most preferred. Next in order of preference were tomato and okra. Dolichos, pigeonpea, cowpea and chickpea were least preferred. In multiple choice test, however, cotton and okra were preferred followed by tomato. Parasites were seen visiting these plants very frequently and high parasitism was recorded on these plants. Chick pea, pigeon pea, cowpea and Dolichos were the least preferred food plants. There appears to be some difference in fecundity as affected by some food plants. Exposure on okra, cotton and tomato resulted in higher cocoon production as compared to pigeonpea, Dolichos, cowpea and chickpea. There was, however, no difference in sex-ratio and longevity of the progeny as affected by food plants. This exotic parasite should be released first in crops such as cotton, okra and tomato on whichH. armigera is a very serious pest in India and elsewhere. Contribution No. 140/86 of the Indian institute of Horticultural Research, Bangalore-560 089     

Efficacy of medicinal plant (Andrographis peniculata) extract on aflatoxin production and growth of Aspergillus flavus

The efficacies of four different concentrations (3, 5, 8 and 10 mg/ml) of an aqueous extract of the Andrographis peniculata were tested on growth and aflatoxin production by Aspergillus flavus in liquid SMKY medium. The maximum inhibition of aflatoxin production and growth of A. flavus were marked at 10 mg/ml (i.e. 78.6% aft. B₁ and 75.1% growth). Growth and aflatoxin production were co-related processes.     

Enhanced degradation of gamma-irradiated lignocelluloses by a new xylanolytic Flavobacterium sp. isolated from soil

An extracellular chitosanase produced by Rhodotorula gracilis CFR-1 that catalyses a limited degradation of chitosan with no detectable generation of glucosamine or reducing groups was identified. Ultracentrifugation, polyacrylamide gel electrophoresis and gel permeation studies suggest that chitosan of average molecular mass 36000 Da was reduced by the enzymic catalysis to nearly one-fourth this size without further hydrolysis of the products. The enzyme, produced constitutively by this yeast, was partially purified and some of its properties were studied.     

Embryology ofMalaxis saprophyta,with comments on the systematic position ofMalaxis (Orchidaceae)

InMalaxis saprophyta, anther wall development corresponds to the Monocotyledonous type. The uninucleate tapetum is of secretory type and the endothecium develops U- and V-shaped thickenings on the inner tangential and radial walls. Cytokinesis is simultaneous; tetrahedral, isobilateral and T-shaped tetrads are formed which are compactly aggregated in pollinia. At anthesis the microspore tetrads are 2-celled. The ovule is anatropous, bitegmic and both integuments are dermal in origin. A single hypodermal cell develops directly into a megaspore mother cell. Embryo sac development is predominantly monosporic and less often bisporic. Irrespective of the type of development, the mature embryo sac is 6-nucleate. Although double fertilization occurs, the primary endosperm nucleus degenerates. Embryogeny is of the Onagrad type. The mature embryo lacks differentiation into cotyledon, plumule and radicle. The reticulate seed coat is formed entirely by the outer layer of outer integument. There are three sterile and three fertile valves in the ovary. Although initially parenchymatous, the entire three sterile valves in the ovary and the upper half of the three fertile valves become sclerified after fertilization. The embryological characters support the disputed systematic position ofMalaxis within subtribeMalaxidinae ofEpidendreae.     

Evaluation of the leaf juice of some higher plants for their toxicity against soil borne pathogens

Out of the leaf juices of eighteen plant species screened, only Eupatorium cannabinum exhibited complete toxicity against Pythium debaryanum, Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii. Shade drying of the leaves had no adverse effect, while oven drying produced an adverse effect on the fungitoxicity of the leaves of E. cannabinum. The crude leaf juice of E. cannabinum successfully inhibited damping-off (Fusarium oxysporum) infection of Pisum sativum seedlings.     

Facial clefts in Saudi Arabia: an epidemiologic analysis in 179 patients

One hundred and seventy-nine consecutive cases of facial clefts that were treated at the King Khalid University Hospital, in Riyadh, Saudi Arabia, were analyzed for an epidemiologic study. Isolated cleft lip was present in 38 percent, cleft of lip and palate in 37.4 percent, and cleft of the posterior palate in only 22.4 percent. There was a male preponderance in all types. In cases of cleft lip with or without cleft palate, the more commonly affected side was the left, followed by bilateral cases. Associated malformations were present in 13.4 percent. A positive family history was found in 26.8 percent of cases. A significant number of patients (7.8 percent) were first seen at more than 10 years of age. The incidence of facial clefts at this hospital was 0.3 per 1000 live births, computed over a period of 6 years. This incidence is significantly lower than that reported from European and Far Eastern countries.     

Shift in product formation from acetate to ethanol using metabolic inhibitors inFusarium oxysporum

Summary Fusarium oxysporum 841 produces a mixture of ethanol and acetic acid from glucose, xylose or Avicel (microcrystalline cellulose) substrates. Some metabolic inhibitors viz. sodium azide, dinitrophenol and polyethylene glycol were used for shifting product formation from acetic acid to ethanol. Using these inhibitors 1.5- to 2- fold increase in ethanol production was achieved with significant repression (by 80 to 90%) of acetic acid. Almost theoretical yields of ethanol were obtained.