Stability of the larvicidal activity of Bacillus thuringiensis subsp. israelensis: amino acid modification and denaturants.

The Bacillus thuringiensis subsp. israelensis mosquito larvicidal toxin is not a sulfhydryl-activated toxin. The protein disulfide bonds were cleaved and blocked without loss of toxicity. In contrast, modification of the lysine side chains eliminated toxicity. Additionally, the toxin was resistant to high concentrations of salt (8 M NaBr), organic solvents (40% methanol), denaturants (4 M urea), and neutral detergents (10% Triton X-100). However, it was inactivated by both positively and negatively charged detergents and by guanidine hydrochloride.     

Calmodulin levels in the yeast and mycelial phases of Ceratocystis ulmi.

The calmodulin content of the yeast and mycelial phases of Ceratocystis ulmi was determined by radioimmunoassay. Calmodulin levels increased at the G1-S boundary of the cell cycle, coinciding with the first visible appearance of buds or germ tubes. However, in both phases the cellular calmodulin levels were equivalent. No differential synthesis was observed.     

Male and female flowers of the dioecious plant sorrel show different patterns of MADS box gene expression.

Male and female flowers of the dioecious plant sorrel (Rumex acetosa) each produce three whorls of developed floral organs: two similar whorls of three perianth segments and either six stamens (in the male) or a gynoecium consisting of a fertile carpel and two sterile carpels (in the female). In the developing male flower, there is no significant proliferation of cells in the center of the flower, in the position normally occupied by the carpels of a hermaphrodite plant. In the female flower, small stamen primordia are formed. To determine whether the organ differences are associated with differences in the expression of organ identity genes, cDNA clones representing the putative homologs of B and C function MADS box genes were isolated and used in an in situ hybridization analysis. The expression of RAD1 and RAD2 (two different DEFICIENS homologs) in males and females was confined to the stamen whorl; the lack of expression in the second, inner perianth whorl correlated with the sepaloid nature of the inner whorl of perianth segments. Expression of RAP1 (a PLENA homolog) occurred in the carpel and stamen whorls in very young flower primordia from both males and females. However, as soon as the inappropriate set of organs ceased to develop, RAP1 expression became undetectable in those organs. The absence of expression of RAP1 may be the cause of the arrest in organ development or may be a consequence.     

Purification and properties of a membrane-bound insulin binding protein, a putative receptor, from Neurospora crassa.

The protein that is responsible for specific, high-affinity binding of insulin to the surface of Neurospora crassa cells has been purified to homogeneity. The insulin binding activity of solubilized plasma membranes resembled that of intact cells with regard to affinity of binding, specificity for mammalian insulins, and amount of insulin bound per cell. Insulin binding activity was purified from Triton X-100 solubilized membranes in two steps: FPLC on a MonoQ HR5/5 column; and affinity chromatography on insulin-agarose. The pure material migrated as a single band of ca. 66 kDa on SDS gels, pI = 7.4 by isoelectric focusing. The protein bound 5.34 pmol of insulin/micrograms, or 35% of that expected for univalent binding. Cross-linking of 125I-insulin to pure protein or to solubilized membranes revealed a single labeled band of 67-70 kDa on SDS gels. In nonreducing native gels, two labeled bands of ca. 55 and 110 kDa were produced after cross-linking, and two bands of similar molecular weight bound iodinated insulin after transfer to nitrocellulose filters. These may correspond to active monomer and dimer forms. The pure protein possessed no protein kinase activity against itself, or against exogenous substrates (histone H2, casein, or the synthetic peptide Glu80-Tyr20), and possessed no detectable phosphorylated amino acids. It is suggested, however, that this 66-kDa protein is the "receptor" that mediates insulin-induced downstream metabolic effects.     

Adenosine deaminase polymorphism among the Semai, Temuan, Semelai, and Jakun groups of West Malaysian Orang Asli

812 West Malaysian Orang Asli belonging to four ethnic groups were surveyed for adenosine deaminase (ADA; EC 3.5.4.4) using starch gel electrophoresis. Only the common ADA1 and ADA2 alleles were found, with the frequencies of the latter being 0.025, 0.103, 0.115 and 0.028 in the Semai, Semelai, Temuan, and Jakun groups, respectively. A new 'breeding genetic distance' was applied to these gene frequencies and the Semelai and Temuan were found to be more closely related to each other, and to have considerably more evolutionary flexibility on this scale of 'micro-evolution' than the other two groups. The Semai and Jakun were more similar to each other on the basis of these ADA gene frequencies.     

Molecular docking analysis of compounds from Lycopersicon esculentum with the insulin receptor to combat type 2 diabetes

It is known that tomato (Lycopersicon esculentum) contains bioactive compounds to combat type-2 diabetes. Therefore, it is of interest to document data from the molecular docking analysis of compounds from Lycopersicon esculentum with the insulin receptors to combat type-2 diabetes. We report the binding features of cinnamic acid, chlorogenic acid, gallic acid & glucoside with insulin receptors for further consideration.     

Arbuscular mycorrhizal and septate endophyte fungal associations in lycophytes and ferns of south India

We studied extent and type of arbuscular mycorrhizal (AM) and septate endophytic (SE) fungal associations in five lycophytes and 50 ferns collected from Eastern and Western Ghats regions. Of the 54 species and one variety (belonging to 31 genera) examined; 54 taxa had AM association and AM fungal structures were absent in Marsilea quadrifolia. This is the first report of AM and SE fungal status for 26 species each. Of the 55 taxa examined, AM morphology has been evaluated for the first time in 51 species. The hydrophytic fern Salvinia molesta was mycorrhizal and non-mycorrhizal at different sites. All the epiphytic and saxicolous species examined were mycorrhizal. The percentage of AM colonization ranged from 22.23 (Christella parasitica) to 82.20 (Adiantum lunulatum) in ferns and 53.46 (Selaginella bryopteris) to 84.34 (Selaginella sp.) in lycophytes. Epiphytic life-forms had the maximum average AM colonization levels, whereas aquatic life-forms had the minimum colonization levels. The percentage root length colonized by septate fungi ranged between 0.59 in Ophioglossum reticulatum and 16.36 in Pteris pellucida. The root length with AM and SE fungal structures as well as their total colonization significantly varied among the taxa examined. Most of the lycophytes and ferns had intermediate-type of AM morphology with a few exhibiting Paris-type. AM fungal spore numbers ranged from 1.0 (Angiopteris evecta, Pteridium aquilinum) to (Nephrolepis exaltata) 9.3 spores per 25 g soil and varied significantly among taxa. AM fungal spore morphotypes belonging to Claroideoglomus, Funneliformis, Glomus and Rhizophagus were recorded.     

Multiple-Herbicide Resistance Is Widespread in Roadside Palmer Amaranth Populations

Herbicide-resistant Palmer amaranth is a widespread issue in row-crop production in the Midsouthern US. Palmer amaranth is commonly found on roadside habitats in this region, but little is known on the degree of herbicide resistance in these populations. Herbicide resistance in roadside Palmer amaranth populations can represent the spread of an adaptive trait across a selective landscape. A large-scale survey was carried out in the Mississippi Delta region of eastern Arkansas to document the level of resistance in roadside Palmer amaranth populations to pyrithiobac and glyphosate, two important herbicides with broad history of use in the region. A total of 215 Palmer amaranth populations collected across 500 random survey sites were used in the evaluations. About 89 and 73% of the surveyed populations showed >90% survival to pyrithiobac and glyphosate, respectively. Further, only 3% of the populations were completely susceptible to glyphosate, while none of the populations was completely controlled by pyrithiobac. Among the 215 populations evaluated, 209 populations showed multiple resistance to both pyrithiobac and glyphosate at varying degrees. Dose-response assays confirmed the presence of high levels of herbicide resistance in the five selected populations (≥ 25-fold compared to a susceptible standard). Results demonstrate the prevalence of multiple-herbicide resistance in roadside Palmer amaranth populations in this region. Growers should be vigilant of Palmer amaranth infestation in roadsides adjacent to their fields and implement appropriate control measures to prevent likely spread of herbicide resistance into their fields.     

Maintenance of NF-kappaB activation in T-lymphocytes and a naive T-cell population in autoimmune-prone (NZB/NZW)F(1) mice by feeding a food-restricted diet enriched with n-3 fatty acids

We have previously shown that feeding a fish oil (FO) supplemented diet in combination with 40% food restriction (FO/FR) has a greater impact on extending life span in lupus-prone (NZB x NZW)F1 mice than either FO ad libitum (FO/AL) or corn oil food restricted (CO/FR) alone. Lupus disease is associated with increased Th-2 (i.e., IL-6 and IL-10) cytokine production and reduced IL-2 production and NF-kappaB activation. We hypothesized that the mechanism of action by which FO/FR increases life span may involve alterations in T-lymphocyte signaling and subsequent cytokine production. To test this hypothesis, we isolated and then stimulated splenic T-lymphocytes ex vivo with anti-CD3 and -CD28 monoclonal antibodies. We report here that CO/FR and FO/FR and to a lesser extent FO/AL offset disease-associated losses in Th-1 cytokine production, CD69 expression, and NF-kappaB activation in splenic T-lymphocytes activated ex vivo. Similarly, CO/FR and FO/FR prevented the disease-dependent rise in Th-2 cytokine production ex vivo and CD69 expression in vivo. In essence, the T-lymphocyte phenotype in the old CO/FR and FO/FR groups was identical to that in the young disease-free mice. Taken together, the data suggest that both CO/FR and FO/FR increase life span, in part, by maintaining a youthful immune phenotype in autoimmune-prone mice. However, FO/FR appears to represent a more potent dietary strategy in delaying disease-associated immune dysregulation than CO/FR.