Biochemical and biologic characterization of exosomes and microvesicles as facilitators of HIV-1 infection in macrophages

Exosomes and microvesicles (MV) are cell membranous sacs originating from multivesicular bodies and plasma membranes that facilitate long-distance intercellular communications. Their functional biology, however, remains incompletely understood. Macrophage exosomes and MV isolated by immunoaffinity and sucrose cushion centrifugation were characterized by morphologic, biochemical, and molecular assays. Lipidomic, proteomic, and cell biologic approaches uncovered novel processes by which exosomes and MV facilitate HIV-1 infection and dissemination. HIV-1 was "entrapped" in exosome aggregates. Robust HIV-1 replication followed infection with exosome-enhanced fractions isolated from infected cell supernatants. MV- and exosome-facilitated viral infections are affected by a range of cell surface receptors and adhesion proteins. HIV-1 containing exosomes readily completed its life cycle in human monocyte-derived macrophages but not in CD4(-) cells. The data support a significant role for exosomes as facilitators of viral infection.     

Acaricidal activity of Cassia alata against Rhipicephalus (Boophilus) annulatus

Using adult immersion test, the acaricidal activity of ethanolic extracts of leaves of Cassia alata L. was studied against Rhipicephalus (Boophilus) annulatus. The efficacy was assessed by measuring per cent adult mortality, inhibition of fecundity and hatching rate. The ethanolic extract of C. alata produced a concentration dependant increase in the adult tick mortality. The highest mortality (45.8%) and inhibition of fecundity (10.9%) were observed at the highest concentration tested (100 mg/ml). The plant extract did not affect egg hatchability.     

Genetic relationship,population structure analysis and pheno-molecular characterization of rice (Oryza sativa L.) cultivars for bacterial leaf blight resistance and submergence tolerance using trait specific STS markers

Rice is an important source of calorie for the growing world population. Its productivity, however is affected by climatic adversities, pest attacks, diseases of bacterial, viral and fungal origin and many other threats. Developing cultivars that are high yielding and stress resilient seems a better solution to tackle global food security issues. This study investigates the potential resistance of 24 rice cultivars against Xanthomonas oryzae pv. Oryzae (Xoo) infection that causes bacterial leaf blight disease and submergence stress. Bacterial leaf blight (BLB) resistance genes (Xa4, xa5, xa13, Xa21, Xa38) and submergence tolerance (Sub1) gene specific markers were used to determine the allelic status of genotypes. The results displayed presence of Xa4 resistance allele (78.95%), xa5 (15.79%) but xa13 and Sub1 tolerance allele were not found in any genotype. However, a new allele for Xa21 (84.21%) and Xa38 (10.52%) were identified in several genotypes. Phenotypic screening for both stress conditions was done to record the cultivars response. None of the genotypes showed resistance against Xoo, although varieties viz., Tapaswini and Konark showed moderate susceptibility. Likewise, survival percentage of genotypes under submergence stress varied from 0 to 100%. Tolerant checks FR13A (100%) and Swarna Sub1 (97.78%) exhibited high survival rate, whereas among genotypes, Gayatri (57.78%) recorded high survivability even though it lacked Sub1 tolerant its genetic background. A total of six trait specific STS and two SSR markers generated an average of 2.38 allele per locus. Polymorphism information content (PIC) value ranged from 0.08 to 0.42 with an average of 0.20. Structure analysis categorized 24 genotypes into two sub-populations, which was in correspondence with Nei’s genetic distance-based NJ tree and principal co-ordinate analysis (PCoA). Swarna Sub1 could be differentiated clearly from BLB resistant check, IRBB60 and other 22 genotypes without having Sub1 gene. Analysis of molecular variance (AMOVA) revealed more genetic variation within population than among population. Principal component analysis (PCA) showed that 9 morphological traits collectively explained 76.126% of total variation among all the genotypes studied. The information from this study would be useful in future breeding programs for pyramiding trait specific genes into high yielding cultivars that fall behind with respect to stress resilience. Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-00951-1.     

Study of LDH variants amongst the Mundas in Ranchi district,Bihar, India

Summary LDH variants were tested for in 392 bloods samples taken from Christian and non-Christian Munda groups, of Ranchi, Bihar by the filter paper technique. Six cases of LDH Cal-1 variant were detected (1.53%). This result is compared with that obtained in Hindu upper caste and Hindu lower caste of West Bengal, and the Munda population was found to be closer to the lower caste people than the upper caste (t=1.752, 0.05> P>0.025).

---

Zusammenfassung 392 Blutproben von Mundas aus christlichen und nichtchristlichen Gruppen, aus Ranchi (Bihar) wurden mit Hilfe der Filterpapiertechnik gesammelt und auf LDH-Varianten gestestet. 6 Fälle (1,53%) der Variante LDH Cal-1 wurden entedeckt. Dieser Wert wird verglichen mit dem von Hindus höherer und niederer Kasten aus West-Bengalen. Die Mundas ähneln den unteren Kasten mehr als den höheren (t=1,752; 0,05> P>0,025).

     

Antibacterial and antilarval-settlement potential and metabolite profiles of novel sponge-associated marine bacteria

In this study, we screened seven novel sponge-associated marine bacteria for their antibacterial and antilarval-settlement activity in order to find possible new sources of non-toxic or less toxic bioactive antifoulants. The anti-bacterial-growth activity of crude extracts of each bacterium was evaluated by the disk-diffusion assay. Extracts of four potent bacteria with high and broad spectra of antibacterial activity were further separated with solvents of different polarities (hexane and ethyl acetate). To evaluate their indirect inhibitive effect on larval settlement, we tested for their antibiofilm formation activity against two of the test bacteria (Vibrio halioticoli and Loktanella hongkongensis) inductive to Hydroides elegans larval settlement. About 60 and 87% of the extracts inhibited biofilm formation by V. halioticoli and by L. hongkongensis respectively. The extracts were also tested for their direct antilarval-settlement activity against the barnacle Balanus amphitrite and the polychaete H. elegans; 87% of the extracts had a strong inhibitive effect on larval settlement of both species. Extracts of two of the isolates completely inhibited larval settlement of B. amphitrite at 70 μg ml⁻¹ and H. elegans at 60 μg ml⁻¹. The organic extracts of Winogradskyella poriferorum effectively inhibited the larval settlement of both H. elegans and B. amphitrite and the biofilm formation of the two bacterial species. The metabolites present in the active crude extracts were profiled using GC MS, and the most prevalent metabolites present in all extracts were identified. This study successfully identified potential new sources of antifouling compounds.

Hemoglobinopathies, G6PD deficiency, and hereditary elliptocytosis in Bahrain

The native population of Bahrain has a high prevalence of hemoglobinopathies and G6PD deficiency, probably as a result of past malarial endemism. We used the Biorad-Variant hemoglobin testing system for primary screening of hemoglobinopathies in 20,000 individuals. Hemoglobin abnormalities were detected in 7,206 (36.3%) cases.     

Separation methods that are capable of revealing blood-brain barrier permeability

The objective of this review is to emphasize the application of separation science in evaluating the blood-brain barrier (BBB) permeability to drugs and bioactive agents. Several techniques have been utilized to quantitate the BBB permeability. These methods can be classified into two major categories: in vitro or in vivo. The in vivo methods used include brain homogenization, cerebrospinal fluid (CSF) sampling, voltametry, autoradiography, nuclear magnetic resonance (NMR) spectroscopy, positron emission tomography (PET), intracerebral microdialysis, and brain uptake index (BUI) determination. The in vitro methods include tissue culture and immobilized artificial membrane (IAM) technology. Separation methods have always played an important role as adjunct methods to the methods outlined above for the quantitation of BBB permeability and have been utilized the most with brain homogenization, in situ brain perfusion, CSF sampling, intracerebral microdialysis, in vitro tissue culture and IAM chromatography. However, the literature published to date indicates that the separation method has been used the most in conjunction with intracerebral microdialysis and CSF sampling methods. The major advantages of microdialysis sampling in BBB permeability studies is the possibility of online separation and quantitation as well as the need for only a small sample volume for such an analysis. Separation methods are preferred over non-separation methods in BBB permeability evaluation for two main reasons. First, when the selectivity of a determination method is insufficient, interfering substances must be separated from the analyte of interest prior to determination. Secondly, when large number of analytes is to be detected and quantitated by a single analytical procedure, the mixture must be separated to each individual component prior to determination. Chiral separation in particular can be essential to evaluate the stereo-selective permeation and distribution of agents into the brain. In conclusion, the usefulness of separation methods during BBB permeability evaluation is immense and more application of these methods is foreseen in the future.