Activation of integrin alphaIIbbeta3 by modulation of transmembrane helix associations

Transmembrane helices of integrin alpha and beta subunits have been implicated in the regulation of integrin activity. Two mutations, glycine-708 to asparagine-708 (G708N)and methionine-701 to asparagine-701, in the transmembrane helix of the beta3 subunit enabled integrin alphaIIbbeta3 to constitutively bind soluble fibrinogen. Further characterization of the G708N mutant revealed that it induced alphaIIbbeta3 clustering and constitutive phosphorylation of focal adhesion kinase. This mutation also enhanced the tendency of the transmembrane helix to form homotrimers. These results suggest that homomeric associations involving transmembrane domains provide a driving force for integrin activation. They also suggest a structural basis for the coincidence of integrin activation and clustering.     

Host Plant-Based Artificial Diets Enhance Development,Survival and Fecundity of the Edible Long-Horned Grasshopper Ruspolia differens (Orthoptera: Tettigoniidae)

Wild swarms of the long-horned grasshoppers Ruspolia differens (Serville) which are widely harvested for consumption and sale in Africa are seasonal and unsustainable, hence the need for innovative ways of artificially producing the insects. We investigated the development, survival, and reproduction of R. differens in the laboratory on diets mixed with host plants [Digitaria gayana Kunth, Cynodon dactylon (L.) and Megathyrsus maximus Jacq (Poales: Poaceae); Ageratum conyzoides L. (Asterales: Asteraceae)] identified from guts of their wild conspecifics with a view to developing a suitable diet for artificial mass rearing of the edible insect. A standard diet comprising ground black soldier fly, Hermetia illucens L. (Diptera: Startiomyidae) larvae, soybean flour, maize flour, vitamin premix, and ground bones was tested for rearing R. differens as a control against the same ingredients incorporated with individual powders of the different host plants. Whereas R. differens developed more slowly in the diet mixed with D. gayana than in the control diet; its development was faster in the diet mixed with C. dactylon. Mortalities of R. differens in host plant-based diets were 42.5–52.5%, far lower than in the control diet with 71% mortality. The insects raised on the diet mixed with M. maximus laid approximately twice more eggs compared to R. differens fecundities from the rest of the diets. However, inclusion of host plants in the diets had no detectable influence on R. differens adult weight and longevity. These findings support inclusion of specific host plants in artificial diets used for mass rearing of R. differens to enhance its survival, development, and fecundity.     

Monitoring biochemical changes in bacterial spore during thermal and pressure-assisted thermal processing using FT-IR spectroscopy

Pressure-assisted thermal processing (PATP) is being widely investigated for processing low acid foods. However, its microbial safety has not been well established and the mechanism of inactivation of pathogens and spores is not well understood. Fourier transform infrared (FT-IR) spectroscopy was used to study some of the biochemical changes in bacterial spores occurring during PATP and thermal processing (TP). Spore suspensions (approximately 10(9) CFU/mL of water) of Clostridium tyrobutyricum, Bacillus sphaericus, and three strains of Bacillus amyloliquefaciens were treated by PATP (121 degrees C and 700 MPa) for 0, 10, 20, and 30 s and TP (121 degrees C) for 0, 10, 20, and 30 s. Treated and untreated spore suspensions were analyzed using FT-IR in the mid-infrared region (4000-800 cm(-1)). Multivariate classification models based on soft independent modeling of class analogy (SIMCA) were developed using second derivative-transformed spectra. The spores could be differentiated up to the strain level due to differences in their biochemical composition, especially dipicolinic acid (DPA) and secondary structure of proteins. During PATP changes in alpha-helix and beta-sheets of secondary protein were evident in the spectral regions 1655 and 1626 cm(-1), respectively. Infrared absorption bands from DPA (1281, 1378, 1440, and 1568 cm(-1)) decreased significantly during the initial stages of PATP, indicating release of DPA. During TP changes were evident in the bands associated with secondary proteins. DPA bands showed little or no change during TP. A correlation was found between the spore's Ca-DPA content and its resistance to PATP. FT-IR spectroscopy could classify different strains of bacterial spores and determine some of the changes occurring during spore inactivation by PATP and TP. Furthermore, this technique shows great promise for rapid screening PATP-resistant bacterial spores.     

Responses of field grown tomato plants to arbuscular mycorrhizal fungal colonization under varying intensities of drought stress

The effects of root colonization by the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck and Smith on growth, flower and fruit production, and fruit quality were studied in field-grown tomato plants exposed to varying intensities of drought stress. Inoculated (M+) and non-inoculated (M−) tomato seedlings were exposed to varying intensities of drought stress by adjusting irrigation intervals. Mycorrhizal plants had significantly higher uptake of N and P in both roots and shoots regardless of intensities of drought stress. AM inoculation also significantly increased shoot dry matter and the number of flowers and fruits. The fruit yields of M+ plants under severe, moderate, mild drought-stressed conditions were higher than M− plants by 24.7%, 23.1%, 16.2% and 12.3%, respectively. Furthermore, M+ plants produced tomato fruits that contain significantly higher quantities of ascorbic acid and total soluble solids (TSS) than M− plants. Mycorrhizal effects increased with increasing intensity of drought. The overall results suggest that mycorrhizal colonization affects host plant nutritional status, water stratus and growth under field conditions and thereby alters reproductive behaviour, fruit production and quality of fruits under both well-watered and drought-stressed conditions.     

Changes in protein synthesis during drought conditioning in roots of jack pine seedlings (Pinus banksiana Lamb.)

The impact of drought conditioning on the ability of eight-week-old jack pine (Pinus banksiana Lamb.) seedlings to withstand drought was assessed. Two progressive cycles of drought conditioning significantly increased the survival of seedlings subjected to a subsequent prolonged drought. The in vivo accumulation of several root membrane proteins during drought conditioning was correlated with an increase in seedling survival. A group of root proteins, ranging in molecular mass from 43 to 47 kDa, increased accumulation during one cycle of drought conditioning and to a lesser extent during two cycles of drought conditioning. The accumulation of several low molecular mass membrane and soluble proteins also increased during drought conditioning, suggesting that these proteins may play an important role in the enhancement of drought tolerance. In vitro translation studies showed a general increase in the abundance of protein products encoded by mRNAs from drought-conditioned seedlings. Although the majority of the in vitro translation products appeared in both control and drought-conditioned seedlings, one mRNA encoding a 15 kDA translated protein was more prominent during the second cycle of drought conditioning.     

Neurological trypanosomiasis in quinapyramine sulfate-treated horses—a breach of the blood–brain barrier?

Trypanosoma evansi infection typically produces wasting disease, but it can also develop into a neurological or meningoencephalitis form in equids. Trypanosomiasis in horses was treated with quinapyramine sulfate, and all the 14 infected animals were recovered clinically. After clinical recovery, four animals developed a neurological form of the disease at various intervals. Two of these animals treated with diminazene aceturate recovered temporarily. Repeated attempts failed to find the parasite in the blood or the cerebrospinal fluid (CSF), but all of the animals were positive in enzyme-linked immunosorbent assay. The calculation of the antibody index (AI) in the serum and the CSF and polymerase chain reaction (PCR) analysis of the CSF and brain tissue were carried out to confirm the neuro-infection. We found PCR and AI analyses of the CSF to be useful tools in the diagnosis of the neurological form of trypanosomiasis when the organism cannot be found in the blood or CSF. The increased albumin quotient is indicative of barrier leakage due to neuroinflammation. The biochemical changes in the CSF due to nervous system trypanosomiasis include increases in the albumin quotient, total protein, and urea nitrogen. It seems to be the first report on relapse of the nervous form of trypanosomiasis in equids even after quinapyramine treatment in endemic areas.     

Toxicity of diafenthiuron to honey bees in laboratory,semi‐field and field conditions

BACKGROUND: Cardamom, an important spice crop often attacked by many insect pests, is controlled mainly using synthetic insecticides. As honey bees play a vital role in pollination in cardamom, the impact of insecticides on honey bees needs to be explored to assess its safety. RESULTS: Risk assessment based on contact toxicity revealed diafenthiuron to be a non‐selective insecticide to bees with a low selectivity ratio (the ratio between the LD₅₀ for beneficial and pest species). A dose of diafenthiuron that killed 90% of cardamom borer, Conogethes punctiferalis Guenee, was found to kill 100% of Indian bees. Based on the hazard ratio (the ratio between the field‐recommended dose and the LD₅₀ for the beneficial), diafenthiuron was found to be slightly to moderately toxic to bees. Diafenthiuron, even at low concentrations of LC₁ (the concentration that killed 1% of bees), was found to affect the foraging and homing behaviour of Indian bees. Of bees fed with 30 µg mL^?1 of diafenthiuron, 40% were found missing on the third day after exposure. However, diafenthiuron did not affect bee visits to the cardamom fields. CONCLUSION: Diafenthiuron is more highly toxic to Apis cerana indica F. than to C. punctiferalis by contact, using selectivity ratio and probit substitution methods of risk assessment, but the hazard ratio revealed diafenthiuron to be a slightly to moderately toxic chemical. Diafenthiuron was found to affect the foraging and homing behaviour of bees at sublethal concentrations. Thus, sublethal effects are more relevant in risk assessment than lethal and acute effects. Copyright © 2010 Society of Chemical Industry