The determination of the antibodies and the somatic O-antigen of Salmonella group B by using the complement-bound lysis of liposomes]

A simple and sensitive method for the determination of group B Salmonella O-antigen and specific antibodies to group B Salmonella by means of the complement-dependent lysis of liposomes sensitized with S. typhimurium lipopolysaccharide (LPS) is proposed. The factors affecting the sensitivity of the method during the determination of antibodies and free antigen have been studied. The method permits the determination of soluble LPS antigen in concentrations of 0.5-200 micrograms/ml.     

Molecular evidence of the survival of subterranean amphipods (Arthropoda) during Ice Age underneath glaciers in Iceland

Two endemic groundwater arthropod crustacean species, Crangonyx islandicus and Crymostygius thingvallensis, were recently discovered on the mid‐Atlantic volcanic island of Iceland. The extent of morphological differences from closest relatives, endemism, along with the geographic isolation of Iceland and its complete coverage by glaciers 21 000 years ago, suggests that these two species have survived glaciation periods in sub‐glacial refugia. Here we provide strong support for this hypothesis by an analysis of mitochondrial genetic variation within Crangonyx islandicus. Our results show that the species is divided into several distinct monophyletic groups that are found along the volcanic zone in Iceland, which have been separated by 0.5 to around 5 million years. The genetic divergence between groups reflects geographic distances between sampling sites, indicating that divergence occurred after the colonization of Iceland. The genetic patterns, as well as the dependency of genetic variation on distances from the tectonic plate boundary and altitude, points to recent expansion from several refugia within Iceland. This presents the first genetic evidence of multicellular organisms as complex as crustacean amphipods which have survived glaciations beneath an ice sheet. This survival may be explained by geothermal heat linked to volcanic activities, which may have maintained favourable habitats in fissures along the tectonic plate boundary in Iceland during glaciations.     

Change in the dynamics of a spin probe in complement-dependent lysis of a liposome

The mobility of phospholipid chains in membranes of liposomes consisting of egg lecitin, cholesterol, dicetylphosphate, sensitized by the lipopolysaccharide antigen F. tularensis by the action of a homologous antiserum and a rabbit complement preparation was studied using 5- and 16-doxylstearate spin probes. It was shown that, during the immune lysis of liposome membranes, changes in the dynamics of spin probes occur, which correlate with the formation of transmembrane channels and exit of the fluorescent marker from the interior of liposomes. It was found that the ratio of the intensities I1/I2 of two low-field extrema in the ESR spectrum is most sensitive to changes in the liposome membrane that are induced by immune components.     

APOE ε4 moderates abnormal CSF-abeta-42 levels,while neurocognitive impairment is associated with abnormal CSF tau levels in HIV+ individuals – a cross-sectional observational study

Background

Cerebrospinal fluid (CSF) biomarkers Aβ1-42, t-tau and p-tau have a characteristic pattern in Alzheimer’s Disease (AD). Their roles in HIV-associated neurocognitive disorder (HAND) remains unclear.

Methods

Adults with chronic treated HIV disease were recruited (n = 43, aged 56.7 ± 7.9; 32% aged 60+; median HIV duration 20 years, >95% plasma and CSF HIV RNA <50 cp/mL, on cART for a median 24 months). All underwent standard neuropsychological testing (61% had HAND), APOE genotyping (30.9% carried APOE ε4 and 7.1% were ε4 homozygotes) and a lumbar puncture. Concentrations of Aβ1-42, t-tau and p-tau were assessed in the CSF using commercial ELISAs. Current neurocognitive status was defined using the continuous Global Deficit Score, which grades impairment in clinically relevant categories. History of HAND was recorded. Univariate correlations informed multivariate models, which were corrected for nadir CD4-T cell counts and HIV duration.

Results

Carriage of APOE ε4 predicted markedly lower levels of CSF Aβ1-42 in univariate (r = -.50; p = .001) and multivariate analyses (R² = .25; p < .0003). Greater levels of neurocognitive impairment were associated with higher CSF levels of p-tau in univariate analyses (r = .32; p = .03) and multivariate analyses (R² = .10; p = .03). AD risk prediction cut-offs incorporating all three CSF biomarkers suggested that 12.5% of participants had a high risk for AD. Having a CSF-AD like profile was more frequent in those with current (p = .05) and past HIV-associated dementia (p = .03).

Conclusions

Similarly to larger studies, APOE ε4 genotype was not directly associated with HAND, but moderated CSF levels of Aβ1-42 in a minority of participants. In the majority of participants, increased CSF p-tau levels were associated with current neurocognitive impairment. Combined CSF biomarker risk for AD in the current HIV+ sample is more than 10 times greater than in the Australian population of the same age. Larger prospective studies are warranted.

     

Comprehensive comparative analysis of kinesins in photosynthetic eukaryotes

Background

Kinesins, a superfamily of molecular motors, use microtubules as tracks and transport diverse cellular cargoes. All kinesins contain a highly conserved ~350 amino acid motor domain. Previous analysis of the completed genome sequence of one flowering plant (Arabidopsis) has resulted in identification of 61 kinesins. The recent completion of genome sequencing of several photosynthetic and non-photosynthetic eukaryotes that belong to divergent lineages offers a unique opportunity to conduct a comprehensive comparative analysis of kinesins in plant and non-plant systems and infer their evolutionary relationships.

Results

We used the kinesin motor domain to identify kinesins in the completed genome sequences of 19 species, including 13 newly sequenced genomes. Among the newly analyzed genomes, six represent photosynthetic eukaryotes. A total of 529 kinesins was used to perform comprehensive analysis of kinesins and to construct gene trees using the Bayesian and parsimony approaches. The previously recognized 14 families of kinesins are resolved as distinct lineages in our inferred gene tree. At least three of the 14 kinesin families are not represented in flowering plants. Chlamydomonas, a green alga that is part of the lineage that includes land plants, has at least nine of the 14 known kinesin families. Seven of ten families present in flowering plants are represented in Chlamydomonas, indicating that these families were retained in both the flowering-plant and green algae lineages.

Conclusion

The increase in the number of kinesins in flowering plants is due to vast expansion of the Kinesin-14 and Kinesin-7 families. The Kinesin-14 family, which typically contains a C-terminal motor, has many plant kinesins that have the motor domain at the N terminus, in the middle, or the C terminus. Several domains in kinesins are present exclusively either in plant or animal lineages. Addition of novel domains to kinesins in lineage-specific groups contributed to the functional diversification of kinesins. Results from our gene-tree analyses indicate that there was tremendous lineage-specific duplication and diversification of kinesins in eukaryotes. Since the functions of only a few plant kinesins are reported in the literature, this comprehensive comparative analysis will be useful in designing functional studies with photosynthetic eukaryotes.     

Energization of membrane vesicles from the cells of glycolyzing bacterium Streptococcus faecalis]

Membrane fractions were isolated from Streptococcus faecalis cells of a glycolyzing microorganism, devoid of the respiratory chain, using the methods of osmotic shock of the protoplasts, ultrasonic treatment of the cells and ultrasonic treatment of the protoplasts. All fractions possessed the ATPase activity, the highest activity being observed in the fraction isolated by ultrasonication of the protoplasts. All preparations were estimated with respect to the presence of vesicles, formed by the "inside-out" and "inside-in" membranes, using ATPase as a marker of the membrane orientation. In the membrane fractions obtained by ultrasonication of the protoplasts, the "inside-out" vesicles were prevalent. ATP-dependent energization of the membranes, sensitive to the action of dicyclohexylcarbodiimide and tetrachlorotrifluoromethyl benzimidazole, was demonstrated by measuring the transport of the lipophylic anion of phenyldicarbaundecaborane and aniline naphthalene sulfonate fluorescence.     

Osmotic activity and ionic permeability of membrane vesicles from Streptococcus faecalis and Micrococcus lysodeikticus cells]

Membrane fractions containing osmotically active vesicles with sufficiently low membrane permeability for K+, Na+ and Cl- ions typical for the intact cell membrane were isolated from the cells of the glycolyzing bacterium Streptococcus faecalis. In their osmotic properties and ionic permeability the membrane fractions of S. faecalis were found similar to those of the respiring bacterium Micrococcus lysodeikticus, which are capable of the energy-dependent potassium transport. It may be thus assumed that the S. faecalis fractions obtained may be used to study ionic transport. The removal of proton-dependent ATPase of the S. faecalis membrane preparations did not affect the permeability of membranes for K+ ions which is indicative of different mechanisms of proton and potassium translocation.     

<Emphasis Type="Italic">Aconitum</Emphasis> biotechnology: recent trends and emerging perspectives

The genus Aconitum (consists more than 250 species) is one of the most important clades of highly valued medicinal plants. Aconitum species are very essential in the traditional device of medication and feature excessive business demand in the herbal marketplace. Some of biologically energetic compounds, e.g., aconitine, indaconitine, pseudoacontine, and so on, had been recognized, and new formulations primarily based on those compounds are being produced as rapid rate. This has led to extensive and rather unregulated exploitation of the species in the wild making the genus a threatened group. Conventional breeding and propagation methods have contributed significantly, but these could not meet up with the ever increasing demands of herbal drug industry globally. Biotechnological interventions, therefore, emerge as an alternative approach in terms of higher production and conservation as well. In recent years, several reports have been published on in vitro propagation of various important Aconitum species. However, advanced biotechnological approaches, such as synthetic seed production and hairy root cultures, are still lacking with only a few reports available. The current review presents an updated overview and critical assessment of secondary data concerning the past and recent biotechnological approaches and interventions in genus Aconitum. This review also attempts to provide a detailed account of work explored so far in micropropagation and emphasizes over the areas not attempted yet, which will act as a baseline data as well as valuable information for different stakeholders and researchers working on various aspects of Aconitum biotechnology.     

Dynamics of accumulation of extracellular lipoteichoic acid in Bacillus cereus

The dynamic of accumulation of extracellular lipoteichoic acid (LTA) was studied depending on the growth stage of Bacillus cereus st. 96. A maximum amount of extracellular LTA was detected in the middle of the exponential growth. The quantity of the biopolymer present in the culture medium at the beginning of the stationary growth under conditions of catabolite repression of sporulation and without repression was found to be different. Experimentally increased concentrations of LTA inhibited B. cereus sporulation. Besides, dormant spores of B. cereus st. 96 were found to contain LTA.