Peripheral biochemical marker for organophosphate-induced delayed neurotoxicity.

Neurotoxicesterase (NTE) activity was assayed in platelets of human and mice as well as in the brain of mice in vitro and in vivo. Mipafox, a well known organophosphate, to induce delayed neurotoxicity, at doses of 5, 10 and 15 mg/kg, subcutaneously, was used to examine the relationship between inhibition of brain and platelet NTE activity in mice. It was observed that the platelet NTE activity of mice was less than in humans. The optimum pH for both brain and platelet NTE of mice, and human platelets, was 8. The results indicate that mipafox produces a dose dependent inhibition of brain and platelet NTE activity in vivo and concentration dependent inhibition in vitro. It can be concluded that assay of platelet NTE can be a useful peripheral biochemical marker for organophosphate-induced delayed neurotoxicity.     

Multiple determinants for the high substrate specificity of an angiotensin II-forming chymase from the human heart

Human heart chymase, a chymotrypsin-like serine proteinase that hydrolyzes the Phe8-His9 bond in angiotensin I (Ang I) to yield the octapeptide hormone angiotensin II (Ang II) and His-Leu, is the most specific, efficient Ang II-forming enzyme described. Other mammalian chymases display a much broader substrate specificity. To better define its substrate specificity, we have mapped the extended substrate-binding site of human heart chymase using Ang I analogs. The enzyme has a preference for aromatic amino acids phenylalanine, tyrosine, and tryptophan at the P1 site. At the S2 subsite there is a significant preference for proline over hydrophobic or hydrophilic amino acids. There is no clear preference for hydrophobic or hydrophilic amino acids at the S'1 and S'2 subsites, but an Ang I analog containing a P'1 proline is not hydrolyzed and one with a P'2 proline is hydrolyzed poorly. An increasing reduction in reactivity occurs when the P position amino acids in Ang I are deleted sequentially from the N terminus. An increase or decrease in the length of the His-Leu leaving group also produces a marked decrease in reactivity. No single determinant in Ang I is preeminently required for efficient catalysis, but several factors acting synergistically appear to be important. Thus, we propose that ideal substrates for human heart chymase should contain the structure nXaa-Pro-[Phe, Tyr, or Trp]-Yaa-Yaa, where n greater than or equal to 6; Xaa = any amino acid; Yaa = any amino acid except proline. This structure exists in Ang I and neurotensin, both of which are good substrates for human heart chymase. These findings indicate that the selection of the scissile bond by the extended substrate-binding site of human heart chymase is more restricted than that in other chymases.     

Bacterial SCP from methanol in Kuwait: Product revovery and composition

The rationale behind KISR's involvement in SCP is explained and emphasis towards the use of aerobic thermophilic/thermotolerant methanol-utilizing bacterial cultures is stressed. An attempt is made to correlate different pilot plant harvesting procedures for biomass recovery with product quality. The latter is expressed as crude and true protein content, moisture and ash content, mineral composition of the ash, and amino acids profiles. The most suitable harvesting procedures were found to be those using a mild heat-acid or acid-heat treatment. Both of these procedures gave high recovery efficiencies and suitable product nutrition.     

Preparatory methods for DNA hydrolysis, cytochemistry, immunocytochemistry and ploidy analysis. Their application to automated and routine diagnostic cytopathology

A review is presented of some methods used to prepare cytologic specimens for analytical and/or automated studies, with the steps of the procedures detailed in appendices. The preparation of the cell monolayers required for optimal automated cell image analysis and classification, e.g., by the Cytoscan 110, is discussed, as is the preparation of poly-L-lysine-coated slides used in the production of monolayered specimens. These monolayers, which can be prepared from a variety of specimens, are also useful for cytochemical and immunocytochemical studies and DNA ploidy analysis. For DNA analysis, a modified gallocyanin chrome alum staining procedure is described as a stoichiometric alternative to the time-consuming Feulgen reaction. The hydrolysis technique required by the latter method is also detailed. The freeze-fracturing technique for the enhancement of monoclonal antibody immunocytochemical staining of detectable antigens is described, along with an indirect immunoalkaline phosphatase staining method. The use of enzyme cytochemical reactions for glucose 6 phosphate dehydrogenase and lysosomal naphthylamidase is also presented.     

Regulation of angiotensin II receptors in cultured rat ovarian granulosa cells by follicle-stimulating hormone and angiotensin II

The regulation of ovarian granulosa cell angiotensin II (Ang-II) receptor formation and progesterone secretion by follicle-stimulating hormone (FSH) and Ang-II was studied in cultured cells prepared from hypophysectomized, diethylstilbestrol-treated immature rats. Ang-II receptors (estimated by the specific cell binding of the Ang-II receptor antagonist 125I-[Sar1,Ile8]Ang-II) were present on freshly prepared granulosa cells and increased by over 2-fold (to 2150 binding sites/cell; KD = 0.5 nM) when cultured in serum-free medium for 48 h. FSH prevented the normal increase in Ang-II receptor expression. Maximal FSH-dependent decrease in Ang-II receptors and increase in progesterone secretion occurred at 100 ng/ml FSH. The inhibitory effect of FSH on granulosa cell Ang-II receptor content was partially mimicked by the cAMP analogue 8-bromo-cAMP, since 8-bromo-cAMP suppressed (by 96%) Ang-II receptor content to a greater extent than FSH (by 60%). Granulosa cell Ang-II receptor content was not modified by progesterone or 17 beta-estradiol, but was decreased by testosterone (by 35%). Ang-II also produced a decrease in granulosa cell Ang-II receptor content, but did not modify progesterone secretion or aromatase activity. The effect of Ang-II on granulosa cell Ang-II receptor content was mimicked by the Ca2+ ionophore A23187, but not by the phorbol ester 12-O-tetradecanoylphorbol 13-acetate, suggesting that an elevation of cytosolic Ca2+ may be important for the homologous down-regulation of the Ang-II receptor. These data show homologous and heterologous down-regulation of granulosa cell Ang-II receptors. If these regulatory mechanisms exist in the FSH-sensitive healthy follicle, our findings suggest that in the process of maturation, healthy and dominant follicles may become decoupled from angiotensinergic influences.     

Frequency of acrocentric association in abnormal sexual development in males and females

A significant difference (P less than 0.05) was observed in a chi 2 comparison of DD, GG and DG-DI associations between male hypogonads and females with primary amenorrhea. This difference increased still further (P less than 0.01) when only DD and GG associations were compared between males and females with abnormal sexual development (ASD). Similarly, when normal males and females were compared for DI, TRI, TETRA, DD vs GG and DG vs GG acrocentric chromosome associations, a significant difference (P less than 0.05) was again observed. The sex difference was also apparent in TRI and TETRA acrocentric associations both in abnormal and normal sexual development males and females. These results suggested that probably sex difference (may be hormonal) influences the number and/or type of acrocentric chromosomes involved in association between males and females with ASD and also between normal males and females.     

Phytotoxic Activity of Tenuazonic Acid Isolated from Alternaria alternata (Fr.) Keissler Causing Leaf Blight of Datura innoxia Mill. and its Effect on Host Metabolism

Tenuazonic acid isolated from Alternaria alternata (Fr.) Keissler causing leaf blight of Datura innoxia Mill. showed significant phytotoxic activity when tested on monocot and dicotyledonous plants. The toxin induced chlorosis and necrosis on leaves of D. in noxia, D. stramonium, D. metel , belladonna, cowpea, wheat, rye, cabbage, cauliflower and maize at 200μg/ml and wilting of seedlings of D. innoxia at 100μg/ml concentration. It also caused complete inhibition of root and shoot elongation of germinating seeds of D. innoxia , wheat, rye, green gram and lettuce at 100μg/ml concentration. It was a nonspecific phytoxin and appeared to have significant role during pathogenesis.
Tenuazonic acid did not cause any significant change in the rate of respiration and in sugar, carbohydrate, total phenol and nitrogen contents of D. innoxia leaves. But its treatment on host plant induced 64 % reduction of chlorophyll content in leaves after 72 hours and 40 % reduction in protein content after 24 hours.     

Combinatorial expression patterns of individual TLE proteins during cell determination and differentiation suggest non-redundant functions for mammalian homologs of Drosophila Groucho

The Drosophila protein Groucho is involved in the regulation of cell-determination events during insect neurogenesis and segmentation. A group of mammalian proteins, referred to as transducin-like Enhancer of split (TLE) 1 through 4, share with Groucho identical structures and molecular properties. The aim was to determine whether individual TLE proteins participate in the regulation of cell determination in mammals like their Drosophila counterpart. It is here reported that TLE family members are expressed in combinatorial ways during the in vitro differentiation of mouse P19 embryonic carcinoma cells (a model for neural determination) and rat CFK2 cells (a model for chondrocytic determination). TLE1 is up-regulated and TLE2 and TLE4 are down-regulated to different extents during early stages of differentiation. In contrast, later stages correlate with up-regulation of TLE2 and TLE4, and decreased expression of TLE1. Individual TLE proteins are also expressed in combinatorial as well as complementary patterns during the development of the cerebral cortex and spinal cord of mouse embryos. In particular, TLE1 is robustly expressed in both neural progenitor cells and postmitotic neurons of the outer layers of the cortical plate, whereas TLE4 expression marks preferentially postmitotic neurons of the inner layers. Taken together, these results strongly suggest non-redundant roles for individual TLE proteins during both cell-determination and cell-differentiation events.     

Pteropyrum naufelum (Polygonaceae), a new species from Iraq

Pteropyrum naufelum , a new species from Iraq, is described and illustrated. It is based on specimens collected in 1962 near the Iraqi boarder.