On the propensity of phosphatidylglycerols to form interdigitated phases

We have determined the phase behavior of disaturated phosphatidylglycerols (PGs) of chain lengths n(CH2) = 14-18 at high pH and ionic strength using calorimetry, dilatometry, as well as x-ray diffraction. PGs with n(CH2) = 14 and 16 show thermotropic behavior similar to that of phosphatidylcholines (PCs). The area/lipid obtained in the gel phase is smaller than that reported for PCs despite the expected larger effective headgroup size. This can be explained by the tilting of the PG headgroup out of the bilayer plane, and we provide experimental evidence for a headgroup tilt transition. For distearoyl PG, we further find that the "usual" gel phase coexists with an interdigitated phase, which exhibits a transition from an orthorhombic into a hexagonal chain packing. The total amount of the interdigitated phase depends significantly on the temperature but is found to be largely independent of temperature equilibration time and different sample preparation protocols. Thus, the development of the interdigitated phase appears to be kinetically trapped. The formation of interdigitated phases in PGs at much smaller chain lengths than in PCs is of high relevance to interaction studies with antimicrobial peptides, as it provides a mechanism for the discrimination of membranes composed of different lipid species.     

Floral biology,breeding system and pollination ecology of Justicia betonica L. (Acanthaceae): An assessment of its low reproductive success in West Bengal,India

Besides its importance as an ornamental plant, Justicia betonica L. is also used as a medicinal plant for the treatment of several human disorders. However, the population size and abundance of the plant species are very low in Indian states, including West Bengal. The breeding system and pollination ecology of J. betonica are still unclear. Therefore, some reproductive aspects were investigated in order to provide important information for the sustainability of the species. The flowering duration spanned from December to May, with its peak in February–March. Flower opening time was much earlier (5.00–8.00 a.m.) on a hot day (in April–May) than on a cold day (10.00 a.m.–12.00 p.m. in December–January). The time of anther dehiscence coincided with the flower opening time; however, the stigma became receptive later. The plant species is self-compatible but dependent on pollinators for fruit and seed sets. Members of different insect groups like ants, bees, butterflies, flies, moths and wasps visit the flowers. Among them, Allorhynchium metallicum and Halictus acrocephalus are the primary pollinators (based on the “relative pollinator service”). The plant species showed a significant pollination deficit (coefficient of pollination deficit [D] = 0.32) in the open condition. Larvae of the florivorous moth Gatesclarkeana erotias significantly reduced reproductive success by eating their floral parts, ovules and immature seeds. In conclusion, combining the pollination deficit and the feeding activity of phytophagous insects (moth larvae) may reduce the reproductive fitness of J. betonica in the study regions.     

Germline soma communication mediated by gap junction proteins regulates epithelial morphogenesis

Gap junction (GJ) proteins, the primary constituents of GJ channels, are conserved determinants of patterning. Canonically, a GJ channel, made up of two hemi-channels contributed by the neighboring cells, facilitates transport of metabolites/ions. Here we demonstrate the involvement of GJ proteins during cuboidal to squamous epithelial transition displayed by the anterior follicle cells (AFCs) from Drosophila ovaries. Somatically derived AFCs stretch and flatten when the adjacent germline cells start increasing in size. GJ proteins, Innexin2 (Inx2) and Innexin4 (Inx4), functioning in the AFCs and germline respectively, promote the shape transformation by modulating calcium levels in the AFCs. Our observations suggest that alterations in calcium flux potentiate STAT activity to influence actomyosin-based cytoskeleton, possibly resulting in disassembly of adherens junctions. Our data have uncovered sequential molecular events underlying the cuboidal to squamous shape transition and offer unique insight into how GJ proteins expressed in the neighboring cells contribute to morphogenetic processes.     

Effect of Extracellular Factors on Growth and Dimorphism of Rhizopus oryzae with Multiple Enzyme Synthesizing Ability

Rhizopus oryzae PR7 MTCC 9642 was a dimorphic fungus that showed a regular 90 days cycle of filament (mycelium) to pellet (yeast) transformation through a distinct bottom dwelling intermediate state and the pellets never revert back to filamentous form. Apart from the normal cycle, high temperature (37°C and above) and extreme pH also induced the yeast formation. Among the ions tested, calcium and chloride ions were found to restore the filamentous morphology, even in extreme pH and temperature. Cysteine HCl also played noteworthy role in maintaining mycelial growth even at adverse condition. Immobilized spores showed the appearance of intermediate form instead of typical yeast form even at high temperature. The strain could produce a number of extracellular hydrolytic enzymes like cellulolytic, xylanolytic, pectinolytic and amylolytic enzymes. The pellet and mycelial forms were found to be a better producer of cellulase–lignocellulase enzymes and amylolytic enzymes respectively, which might be correlated with their infectivity. Increase in inoculum size, agitation during cultivation, change in carbon and nitrogen source failed to induce mycelial growth in extreme conditions, which might be explained as irreversible change of configuration of protein responsible for mycelial development.     

Recruitment of HRDC domain of WRN and BLM to the sites of DNA damage induced by mitomycin C and methyl methanesulfonate

The HRDC (helicase and RNase D C-terminal) domain at the C-terminal of WRNp (Werner protein) (1150-1229 amino acids) and BLMp (Bloom protein) (1212-1292 amino acids) recognize laser microirradiation-induced DNA dsbs (double-strand breaks). However, their role in the recognition of DNA damage other than dsbs has not been reported. In this work, we show that HRDC domain of both the proteins can be recruited to the DNA damage induced by MMS (methyl methanesulfonate) and MMC (methyl mitomycin C). GFP (green fluorescent protein)-tagged HRDC domain produces distinct foci-like respective wild-types after DNA damage induced by the said agents and co-localize with γ-H2AX. However, in time course experiment, we observed that the foci of HRDC domain exist after 24 h of removal of the damaging agents, while the foci of full-length protein disappear completely. This indicates that the repair events are not completed by the presence of protein corresponding to only the HRDC domain. Consequently, cells overexpressing the HRDC domain fail to survive after DNA damage, as determined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay. Moreover, 24 h after removal of damaging agents, the extent of DNA damage is greater in cells overexpressing HRDC domain compared with corresponding wild-types, as observed by comet assay. Thus, our observations suggest that HRDC domain of both WRN and BLM can also recognize different types of DNA damages, but for the successful repair they fail to respond to subsequent repair events.     

Galectin-1 signaling in leukocytes requires expression of complex-type N-glycans

Dimeric galectin-1 (dGal-1) is a homodimeric lectin with multiple proposed functions. Although dGal-1 binds to diverse glycans, it is unclear whether dGal-1 preferentially binds to specific subsets of glycans on cell surfaces to transmit signals. To explore this question, we selectively inhibited major glycan biosynthetic pathways in human HL60, Molt-4, and Jurkat cells. Inhibition of N-glycan processing blocked surface binding of dGal-1 and prevented dGal-1-induced Ca(2+) mobilization and phosphatidylserine exposure. By contrast, inhibition of O-glycan or glycosphingolipid biosynthesis did not affect dGal-1 binding or dGal-1-induced Ca(2+) mobilization and phosphatidylserine exposure. These results demonstrate that dGal-1 preferentially binds to and signals through glycoproteins containing complex-type N-glycans in at least some leukocyte subsets.     

Physicochemical studies on goat pulmonary surfactant

The large aggregate (LA) fraction of goat pulmonary surfactant (GPS) was isolated and characterized. Goat lung surfactant extract (GLSE) was obtained by chloroform-methanol extraction of the saline suspended LA fraction. Total phospholipid (PL), cholesterol (CHOL), and protein were biochemically estimated. It was composed of approximately 83% (w/w) PL, approximately 0.6% (w/w) CHOL and approximately 16% (w/w) protein. CHOL content was found to be lower while the protein content was found to be higher than other mammalian pulmonary surfactants. Electrospray Ionization Mass Spectrometry (ESIMS) of GLSE confirmed dipalmitoylphosphatidylcholine (DPPC) as the major phospholipid species, with significant amounts of palmitoyl-oleoyl phosphatidylcholine (POPC), palmitoyl-myristoyl phosphatidylcholine (PMPC) and dioleoylphosphatidylcholine (DOPC). Functionality of the solvent spread GLSE film was carried out in a Langmuir surface balance by way of surface pressure (pi)-area (A) measurements. A high value of pi (approximately 65 mN m(-1)) could be attained with a lift-off area of approximately 1.2 nm(2) molecule(-1). A relatively large hysteresis was observed during compression-expansion cycles. Monolayer deposits at different pi, transferred onto freshly cleaved mica by Langmuir-Blodgett (LB) technique, were imaged by atomic force microscopy. DPPC-enriched domains (evident from height analyses) showed dimensions of 2.5 microm and underwent changes in shapes after 30 mN m(-1). Functionality and structure of the surfactant films were proposed to be controlled by the relative abundances of protein and cholesterol.