A highly potent and safe pyrrolopyridine-based allosteric HIV-1 integrase inhibitor targeting host LEDGF/p75-integrase interaction site

Allosteric integrase inhibitors (ALLINIs) are a class of experimental anti-HIV agents that target the noncatalytic sites of the viral integrase (IN) and interfere with the IN-viral RNA interaction during viral maturation. Here, we report a highly potent and safe pyrrolopyridine-based ALLINI, STP0404, displaying picomolar IC₅₀ in human PBMCs with a >24,000 therapeutic index against HIV-1. X-ray structural and biochemical analyses revealed that STP0404 binds to the host LEDGF/p75 protein binding pocket of the IN dimer, which induces aberrant IN oligomerization and blocks the IN-RNA interaction. Consequently, STP0404 inhibits proper localization of HIV-1 RNA genomes in viral particles during viral maturation. Y99H and A128T mutations at the LEDGF/p75 binding pocket render resistance to STP0404. Extensive in vivo pharmacological and toxicity investigations demonstrate that STP0404 harbors outstanding therapeutic and safety properties. Overall, STP0404 is a potent and first-in-class ALLINI that targets LEDGF/p75 binding site and has advanced to a human trial.     

Novel Anti-Apoptotic MicroRNAs 582-5p and 363 Promote Human Glioblastoma Stem Cell Survival via Direct Inhibition of Caspase 3, Caspase 9, and Bim

Glioblastoma is the most common and lethal primary brain tumor. Tumor initiation and recurrence are likely caused by a sub-population of glioblastoma stem cells, which may derive from mutated neural stem and precursor cells. Since CD133 is a stem cell marker for both normal brain and glioblastoma, and to better understand glioblastoma formation and recurrence, we looked for dys-regulated microRNAs in human CD133+ glioblastoma stem cells as opposed to CD133+ neural stem cells isolated from normal human brain. Using FACS sorting of low-passage cell samples followed by microRNA microarray analysis, we found 43 microRNAs that were dys-regulated in common in three separate CD133+ human glioblastomas compared to CD133+ normal neural stem cells. Among these were several microRNAs not previously associated with cancer. We then verified the microRNAs dys-regulated in glioblastoma using quantitative real time PCR and Taqman analysis of the original samples, as well as human GBM stem cell and established cell lines and many human specimens. We show that two candidate oncogenic microRNAs, miR-363 and miR-582-5p, can positively influence glioblastoma survival, as shown by forced expression of the microRNAs and their inhibitors followed by cell number assay, Caspase 3/7 assay, Annexin V apoptosis/fluorescence activated cell sorting, siRNA rescue of microRNA inhibitor treatment, as well as 3′UTR mutagenesis to show luciferase reporter rescue of the most successful targets. miR-582-5p and miR-363 are shown to directly target Caspase 3, Caspase 9, and Bim.     

The investigation of the combined effects of gamma irradiation and environmental manipulation on mortality and sterility of Rhyzopertha dominica (Fabricius) (Coleoptera: Bostrichidae)

With respect to the limitations use of methyl bromide and phosphine, employing ionizing radiation to control stored product pests has attracted great attention. The aim of this study is the investigation of the combined effects of gamma irradiation as viable alternatives to synthetic insecticides and environmental management on mortality and sterility of Rhyzopertha dominica (Fabricius) in a wheat cultivar (Gascogen). The effect of doses range from 30 to 2000?Gy gamma irradiation in combination with manipulation of temperature (15, 21, 27 and 32?°C) and relative humidity (20%, 50%, 65% and 85%) on 5–10?days old adults of R. dominica in Gascogen cultivar of wheat were explored. The experiments were repeated three times and conducted in The Nuclear Agriculture Research School in Karaj and laboratory of shahid steki silo in shahre-kord. Probit analysis revealed that both temperature and relative humidity had combined effects when used with gamma irradiation. The lowest doses of gamma ray required to kill 25% (14.2?Gy) and 50% (610.8?Gy) of the population (LD₂₅ and LD₅₀) were recorded at 21?°C and 85% relative humidity respectively. The low dose for 99% mortality (LD₉₉; 2386.7?Gy) was recorded for beetles maintained at 21?°C and 50% relative humidity. The effect of temperature (15, 21, 27 and 32?°C) on sterility caused by gamma irradiation was also investigated. The results showed that the F₁ generation emerged only when the beetles were treated with doses of 0–100?Gy at 32?°C and 0–70?Gy at 27?°C. These results indicate that temperature and relative humidity play an important role in the susceptibility of the lesser grain beetle to gamma irradiation. The results suggest that controlling the efficiency of gamma radiation through environmental control allows the use of low doses of gamma radiation that have a less harmful effect on human health, non-target organisms and seed agronomic features.     

Potential of Lentibacillus sp. NS12IITR for production of lipids with enriched branched-chain fatty acids for improving biodiesel properties along with hydrocarbon co-production

Extremophiles - Hypersaline environment is inhabited by array of microbes which have the potential to produce industrially important products. This study explored biomass and lipid production...     

TYPES OF LATICIFERS AND CRYSTALS IN JATROPHA AND THEIR TAXONOMIC IMPLICATIONS

Anatomical examination of 37 species of Jatropha (Euphorbiaceae) has revealed the occurrence of two distinct types of laticifers—articulated, and nonarticulated—in addition to idioblasts that are irregularly shaped individual cells. With the notable exception of J. augustii Pax & K. Hoffm. (section Peltatae (Pax) Dehgan & Webster), these idioblasts occur in the two most advanced sections of the subgenus Curcas (Adans.) Pax, namely Loureira (Cav.) Muell. Arg. ex Pax and Mozinna (Ortega) Pax. The presence of two laticifer types and “idioblastic laticifers” in the genus, in association with the morphological reduction series found with evolutionary advancement are, therefore, significant in the delimitation of sections and subsections. Further evidence is presented by the occurrence of “chambered crystalliferous cells” in subgenus Curcas, but not in subgenus Jatropha (=Adenorhopium Griseb.). Reexamination of laticifers in other genera of the Euphorbiaceae is suggested as a possible means of alleviating the long-standing taxonomic dilemma of this large and morphologically diverse family.     

Critical role of Glu175 on stability and folding of bacterial luciferase: stopped-flow fluorescence study

Bacterial luciferase is a heterodimeric enzyme, which catalyzes the light emission reaction, utilizing reduced FMN (FMNH2), a long chain aliphatic aldehyde and O(2), to produce green-blue light. This enzyme can be readily classed as slow or fast decay based on their rate of luminescence decay in a single turnover. Mutation of Glu175 in alpha subunit to Gly converted slow decay Xenorhabdus Luminescence luciferase to fast decay one. The following studies revealed that changing the luciferase flexibility and lake of Glu-flavin interactions are responsible for the unusual kinetic properties of mutant enzyme. Optical and thermodynamics studies have caused a decrease in free energy and anisotropy of mutant enzyme. Moreover, the role of Glu175 in transition state of folding pathway by use of stopped-flow fluorescence technique has been studied which suggesting that Glu175 is not involved in transition state of folding and appears as surface residue of the nucleus or as a member of one of a few alternative folding nuclei. These results suggest that mutation of Glu175 to Gly extended the structure of Xenorhabdus Luminescence luciferase, locally.     

Nasal cytobrush cytology: evaluation of the hyperchromatic supranuclear stria

OBJECTIVE: To evaluate the hyperchromatic supranuclear stria (SNS) in various types of rhinopathies. STUDY DESIGN: The study included 42 patients with rhinopathies and a control group consisting of 28 healthy adults. The rhinopathy group was categorized into 4 subgroups, including allergic rhinitis, infective rhinitis, vasomotor rhinitis and mixed group. Cytologic samples were obtained by cytobrush from the middle one third of the inferior turbinate. RESULTS: The hyperchromatic SNS was present in the majority of ciliated cells in a high percentage in the control group (91.7%), whereas in the pathologic group it was 40%. The difference is significant (p = 0.0000). CONCLUSION: Nasal cytology is a simple, reliable tool for the diagnosis of rhinopathies.     

The use of proteome similarity for the qualitative and quantitative profiling of reperfused myocardium

An LC-MS-based approach is presented for the identification and quantification of proteins from unsequenced organisms. The method relies on the preservation of homology across species and the similarity in detection characteristics of proteomes in general. Species related proteomes share similarity that progresses from the amino acid frequency distribution to the complete amino sequence of matured proteins. Moreover, the comparative analysis between theoretical and experimental proteome distributions can be used as a measure for the correctness of detection and identification obtained through LC-MS-based schemes. Presented are means to the identification and quantification of rabbit myocardium proteins, immediately after inducing cardiac arrest, using a data-independent LC-MS acquisition strategy. The employed method of acquisition affords accurate mass information on both the precursor and associated product ions, whilst preserving and recording the intensities of the ions. The latter facilitates label-free quantification. The experimental ion density observations obtained for the rabbit sub proteome were found to share great similarity with five other mammalian samples, including human heart, human breast tissue, human plasma, rat liver and a mouse cell line. Redundant, species-homologues peptide identifications from other mammalian organisms were used for initial protein identification, which were complemented with peptide identifications of translated gene sequences. The feasibility and accuracy of label-free quantification of the identified peptides and proteins utilizing above mentioned strategy is demonstrated for selected cardiac rabbit proteins.