Psychosine-induced alterations in peroxisomes of twitcher mouse liver

Krabbe disease is a neuroinflammatory disorder in which galactosylsphingosine (psychosine) accumulates in nervous tissue. To gain insight into whether the psychosine-induced effects in nervous tissue extend to peripheral organs, we investigated the expression of cytokines and their effects on peroxisomal structure/functions in twitcher mouse liver (animal model of Krabbe disease). Immunofluorescence analysis demonstrated TNF-α and IL-6 expression, which was confirmed by mRNAs quantitation. Despite the presence of TNF-α, lipidomic analysis did not indicate a significant decrease in sphingomyelin or an increase in ceramide fractions. Ultrastructural analysis of catalase-dependent staining of liver sections showed reduced reactivity without significant changes in peroxisomal contents. This observation was confirmed by assaying catalase activity and quantitation of its mRNA, both of which were found significantly decreased in twitcher mouse liver. Western blot analysis demonstrated a generalized reduction of peroxisomal matrix and membrane proteins. These observations indicate that twitcher mouse pathobiology extends to the liver, where psychosine-induced TNF-α and IL-6 compromise peroxisomal structure and functions.     

Chromium-tolerant bacteria isolated from industrial effluents and their use in detoxication of hexavalent chromium

Three bacterial strains were isolated from effluents of leather (CMBL Cr13, CMBL Cr14) and steel (CMBL Cr15) industries for their possible use in chromium(VI) detoxication of industrial waste. CMBL Cr13 was found to tolerate chromium(VI) up to a concentration of 45 g/L in the medium, while CMBL Cr14 and CMBL Cr15 could tolerate up to 40 g/L. These bacteria were also checked for resistance to other metals. They resisted a lead concentration of 1 g/L and cadmium concentration of 550 mg/L in the medium. They showed optimum growth at pH 7.3–7.5 at a temperature of 35–37°C. Cr^VI-reducing ability of the three strains ranged from 70 to 80% after 3 d of incubation. The possible use of these bacteria in environmental cleanup is discussed.     

Reactive centre loop mutants of α-1-antitrypsin reveal position-specific effects on intermediate formation along the polymerization pathway

The common severe Z mutation (E342K) of α₁-antitrypsin forms intracellular polymers that are associated with liver cirrhosis. The native fold of this protein is well-established and models have been proposed from crystallographic and biophysical data for the stable inter-molecular configuration that terminates the polymerization pathway. Despite these molecular ‘snapshots’, the details of the transition between monomer and polymer remain only partially understood. We surveyed the RCL (reactive centre loop) of α₁-antitrypsin to identify sites important for progression, through intermediate states, to polymer. Mutations at P₁₄P₁₂ and P₄, but not P₁₀P₈ or P₂P_1′, resulted in a decrease in detectable polymer in a cell model that recapitulates the intracellular polymerization of the Z variant, consistent with polymerization from a near-native conformation. We have developed a FRET (Förster resonance energy transfer)-based assay to monitor polymerization in small sample volumes. An in vitro assessment revealed the position-specific effects on the unimolecular and multimolecular phases of polymerization: the P₁₄P₁₂ region self-inserts early during activation, while the interaction between P₆P₄ and β-sheet A presents a kinetic barrier late in the polymerization pathway. Correspondingly, mutations at P₆P₄, but not P₁₄P₁₂, yield an increase in the overall apparent activation energy of association from ~360 to 550 kJ mol⁻¹.     

Methoprene modulates the effect of diet on male melon fly,Bactrocera cucurbitae,performance at mating aggregations

The effect of access to dietary protein (P) (hydrolyzed yeast) and/or treatment with a juvenile hormone analogue, methoprene (M), (in addition to sugar and water) on male aggregation (lekking) behaviour and mating success was studied in a laboratory strain of the melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae). Six‐day‐old males were treated with (1) protein and methoprene (M+P+), (2) only protein (M?P+), or (3) only methoprene (M+P?), and compared with 14‐day‐old sexually mature untreated males (M?P?). The lekking behaviour of the four groups of males when competing for virgin sexually mature females (14 –16 days old) was observed in field cages. The following parameters were measured at male aggregations: lek initiation, lek participation, males calling, male–male interaction, female acceptance index, and mating success. For all these parameters, the M+P+ males significantly outperformed the other males. Moreover, for all parameters, there was a similar trend with M+P+ > M?P+ > M?P? > M+P?. More M+P+ males called and initiated and participated in lek activities than all other types of male, which resulted in higher mating success. They had also fewer unsuccessful copulation attempts than their counterparts. Whereas treatment with methoprene alone had a negative effect in young males with only access to sugar, access to dietary protein alone significantly improved young male sexual performance; moreover, the provision of methoprene together with protein had a synergistic effect, improving further male performance at leks. The results are of great relevance for enhancing the application of the sterile insect technique (SIT) against this pest species. The fact that access to dietary protein and treatment of sterile males with methoprene improves mating success means that SIT cost‐effectiveness is increased, as more released males survive to sexual maturity.     

Prospect of phytoaccumulation of arsenic by Brassica juncea (L.) in Bangladesh

The phytoaccummulation of arsenic by Brassica juncea (L.) was investigated for varying concentrations selected within the range that is evident in Bangladeshi soil. B. juncea (Rai and BARI-11) was grown in the hydroponic media under greenhouse condition with different concentrations (0.5, 1.0, 15, 30, 50 and 100 ppm) of sodium arsenite. Inductively coupled plasma atomic emission spectroscopy (ICP-AES) was used to analyze the data. Mapping of potential area of phytoaccumulation of arsenic by B. juncea was done using Geographic information system (GIS). Arsenic was detected at lower concentrations (0.5 and 1.0 ppm) only at root system of the plant. For higher concentrations (15, 30, and 50 ppm) arsenic was detected both in the root and shoot systems. The results suggested that at 15 and 50 ppm uptake was higher compared to 30 ppm. For 100 ppm of arsenic no plant growth was observed. In Bangladesh, where concentration of arsenic is at lower level and present only at rooting zone, B. juncea may be used for phytoaccumulation of arsenic keeping usual agronomic practices. However, for higher concentrations, B. juncea can be regarded as a good accumulator of arsenic where uptake of arsenic was up to 1% of total biomass of the plant.     

Adaptation of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> to Thermal Stress Yields a Thermotolerant Variant Which Also Exhibits Improved Survival at pH 2

Loss in probiotic viability upon exposure to stressful storage and transport conditions has plagued the probiotic market worldwide. Lactobacillus acidophilus is an important probiotic that is added to various functional foods. It is known to be fairly labile and susceptible to temperature variations that it encounters during processing and storage which increases production cost. It has been repeatedly demonstrated that pre-exposure to sub-lethal doses of stress, particularly, temperature and pH, leads to improved survival of various probiotics when they subsequently encounter the same stress of a much greater magnitude. Attempts to adapt L. acidophilus to temperatures as high as 65 °C to arrive at a thermotolerant variant have not been reported previously. To improve viability at elevated temperatures, we gradually adapted the L. acidophilus NCFM strain to survival at 65 °C for 40 min. Following adaptation, the variant showed a 2-log greater survival compared to wild-type at 65 °C. Interestingly, this thermotolerant variant also demonstrated a 2-log greater stability compared to wild-type at pH 2.0. The improved pH and temperature stress tolerance exhibited by this variant remained unaltered even when the strain was lyophilized. Moreover, the thermotolerant variant demonstrated improved stability compared to wild-type when stored for up to a week at 37 and 42 °C. Probiotic properties of the variant such as adherence to epithelial cells and antibacterial activity remained unaltered. This strain can potentially help address the issue of significant loss in viable cell counts of L. acidophilus which is typically encountered during probiotic manufacture and storage.     

Improving vanadium stress tolerance of watermelon by grafting onto bottle gourd and pumpkin rootstock

Vanadium (V) is a transition metal found in the Earth crust. V adversely affects plant growth and development. Besides several other management practices, grafting of scion cultivars onto appropriate rootstock provides a suitable solution. Grafting is an important agro-technical procedure utilized to enhance the capacity of plants to tolerate biotic and abiotic stresses. In this study, watermelon was grafted onto bottle gourd and pumpkin rootstock, and self-grafted watermelon plants were utilized as a control. V was applied at the rate of 50 mg/L under hydroponic conditions. The result showed that V application substantially reduces the growth of watermelon plants, however, grafting of watermelon onto bottle gourd and pumpkin rootstock improves V stress tolerance of watermelon by reducing the V concentration in leaf tissues, improving the relative chlorophyll content (SPAD index) and photosynthetic assimilation, up-regulating the expression of SOD (Cla008698, Cla0012125, Cla009820 and Cla001158), glutathione S-transferase (Cla013224) and glutathione peroxidase (Cla021039) genes in the leaves, and enhancing the activities of antioxidant enzymes (SOD, CAT). The scanning electron microscopy (SEM) of the root tips showed that minimal damage of roots was observed for pumpkin roots compared with the roots of watermelon and bottle gourd under V stress conditions. So far as we know, these results are the first evidence that grafting mitigates V stress in plants.     

Modern pollen assemblages from Reasi (Jammu and Kashmir), India: a tool for interpreting fossil pollen records

Twenty five surface samples/moss cushions were collected for palynological analysis from open areas of Reasi District, Jammu and Kashmir (India). These samples were used to investigate the relationships between extant vegetation and modern pollen spectra, which serve as modern analogue for the reliable ecological interpretation of fossil pollen records. The present vegetation in the region comprises tropical dry deciduous forests and subtropical pine forests with scattered stands of oak. The pollen analysis reveals that Pinus sp. (average 69% in the pollen assemblages), amongst the conifers, dominates the pollen rain, which can be attributed to its high pollen productivity and exceptional pollen dispersal efficiency. Cedrus sp. and Podocarpus sp. pollen contribute with an average of 16 and 5% to the total pollen rain. Other conifers such as Picea sp., Abies sp., Juniperus sp. and Tsuga sp., as well as broad-leaved taxa such as Quercus sp., Alnus sp., Betula sp., Carpinus sp., Corylus sp., Juglans sp., Ulmus sp., Salix sp., Elaeocarpus sp., Mallotus sp. and Aesculus sp., have lower averages of 1 to 4.5% in the total pollen rain which could be either due to their poor pollen dispersal efficiency or to the poor preservation in the samples. Tubuliflorae (average 25%), Poaceae (average 6.26%), Cerealia and other crop plants (average 7.68%) are other prominent taxa in the pollen rain. The nearly complete absence of members of tropical dry deciduous forests in the pollen spectra likely is due to the fact that most species in this vegetation type are not wind pollinated.