Proteomic Analysis of the Mycobacterium tuberculosis Outer Membrane for Potential Implications in Uptake of Small Molecules.

Increased resistance to current antimycobacterial agents and a potential bias toward relatively hydrophobic chemical entities highlight an urgent need to understand how current anti-TB drugs enter the tubercle bacilli. While inner membrane proteins are well-studied, how small molecules cross the impenetrable outer membrane remains unknown. Here, we employed mass spectrometry-based proteomics to show that octyl-ß-d-glucopyranoside selectively extracts the outer membrane proteins of Mycobacterium tuberculosis. Differentially expressed proteins between nutrient-replete and nutrient-depleted conditions were enriched to identify proteins involved in nutrient uptake. We demonstrate cell surface localization of seven new proteins using immunofluorescence and show that overexpression of the proteins LpqY and ProX leads to hypersensitivity toward streptomycin, while overexpression of SubI, SpmT, and Rv2041 exhibited higher membrane permeability, assessed through an EtBr accumulation assay. Further, proton NMR metabolomics suggests the role of six outer membrane proteins in glycerol uptake. This study identifies several outer membrane proteins that are involved in the permeation of small hydrophilic molecules and are potential targets for enhancing the uptake and efficacy of anti-TB drugs.

Photoautotrophic black-colored cyanobacterial soil crust biosynthesizes photoprotective compounds and is capable of using blue, green, and red wavelengths of light for its growth.

Several cyanobacteria can adjust their light-harvesting machinery in response to existing light signals in a process called chromatic acclimation (CA) which permits the utilization of available light resources for photosynthesis. CA involves alteration in the pigment composition of a major light-harvesting complex called phycobilisome (PBS) and allows some cyanobacteria to utilize green light (GL) to drive photosynthesis. However, cyanobacteria, in contrast with eukaryotic algae and higher plants, can not utilize blue light (BL) for photosynthesis due to their dependency on PBS. Here, we studied a black-colored soil crust that was composed of a single cyanobacterium identified and named Oscillatoria sp. Malviya-1 after phenotypic and phylogenetic analyses. The black-colored crust can absorb light from almost all parts of photosynthetically active radiation (400-700 nm) and ultraviolet radiation (280-400 nm) due to the presence of photosynthetic pigments and microbial sunscreens such as chlorophyll ɑ, carotenoids, phycoerythrin, phycocyanin, allophycocyanin, mycosporine-like amino acids, and scytonemin. Unlike other cyanobacteria, Oscillatoria sp. Malviya-1 can grow using GL, BL, and red light (RL) in addition to white light (WL) which was accompanied by the different colors of the mat under different light conditions. The presence of CA and sunscreens compounds can maximize the fitness of soil crust under a dynamic light environment, UVR, and desiccation. Detailed study of Oscillatoria sp. Malviya-1 will provide information on the mechanism of CA in cyanobacterial soil crust and its unique ability to use both GL and BL.

Characterization and low-cost preservation of Chromobacterium violaceum strain TRFM-24 isolated from Tripura state, India.

BACKGROUND: Chromobacterium species, through their bioactive molecules, help in combating biotic and abiotic stresses in plants and humans. The present study was aimed to identify, characterize and preserve in natural gums the violet-pigmented bacterial isolate TRFM-24 recovered from the rhizosphere soil of rice collected from Tripura state. RESULTS: Based on morphological, biochemical and 16S rRNA gene sequencing, the isolate TFRM-24 was identified as Chromobacterium violaceum (NAIMCC-B-02276; MCC 4212). The bacterium is saprophytic, free living and Gram negative. The strain was found positive for production of IAA, cellulase, xylanase and protease, and showed tolerance to salt (2.5%) and drought (-1.2 MPa). However, it showed poor biocontrol activity against soil-borne phytopathogens and nutrient-solubilizing abilitiets. C. violaceum strain TRFM-24 did not survive on tryptic soya agar (TSA) beyond 12 days between 4 and 32 °C temperature hence a method of preservation of this bacterium was attempted using different natural gums namely Acacia nilotica (babul), Anogeissus latifolia (dhavda), Boswellia serrata (salai) and Butea monosperma (palash) under different temperature regime (6-32 °C). The bacterium survived in babul gum (gum acacia), dhavda and salai solution at room temperature beyond a year. CONCLUSION: Based on polyphasic approach, a violet-pigmented isolate TRFM-24 was identified as Chromobacterim violaceum which possessed some attributes of plant and human importance. Further, a simple and low-cost preservation method of strain TRFM-24 at room temperature was developed using natural gums such as babul, dhavda and salai gums which may be the first report to our knowledge.

Outcome Analysis of Dual Implant Osteosynthesis for Ipsilateral Proximal and Shaft Femur Fractures: Do We Need Cephalomedullary Nails?

Introduction Most surgeons prefer a single implant for segmental proximal and diaphyseal femur fractures, although results are controversial and still no consensus for proper management is present. This prospective study analyses the functional and radiological outcome of managing 17 patients with ipsilateral shaft and proximal femur fractures by dual implant osteosynthesis at our center. Methods Over a two-year period, we managed 17 patients with a mean age of 35 years, with cancellous cannulated screws or dynamic hip screws for intracapsular femur fractures and improvised proximal femoral nail for extracapsular proximal femur fractures. Distal femoral locking plates or distal femoral nails were used for shaft femur fractures depending upon fracture morphology. The patients had a maximum follow-up of 18 months. Results A total of 80% of patients had a good functional outcome (using the Friedman-Wyman scoring system) while 60% had an excellent Harris Hip Score. The mean time taken for the bone union for proximal femur fractures was 4.75 months and for shaft femur fractures, it was 6 months. Conclusion We had a satisfactory functional and clinical outcome of managing these fractures with two implants, one focusing biomechanically on each fracture. This principle of dual implant osteosynthesis can reliably be used in such difficult fracture patterns and it negates the use of the single cephalomedullary nail for fixating both fractures.

Roadmap to sustainable carbon-neutral energy and environment: can we cross the barrier of biomass productivity?

The total number of inhabitants on the Earth is estimated to cross a record number of 9 × 103 million by 2050 that present a unique challenge to provide energy and clean environment to every individual. The growth in population results in a change of land use, and greenhouse gas emission due to increased industrialization and transportation. Energy consumption affects the quality of the environment by adding carbon dioxide and other pollutants to the atmosphere. This leads to oceanic acidification and other environmental fluctuations due to global climate change. Concurrently, speedy utilization of known conventional fuel reservoirs causes a challenge to a sustainable supply of energy. Therefore, an alternate energy resource is required that can maintain the sustainability of energy and environment. Among different alternatives, energy production from high carbon dioxide capturing photosynthetic aquatic microbes is an emerging technology to clean environment and produce carbon-neutral energy from their hydrocarbon-rich biomass. However, economical challenges due to low biomass production still prevent the commercialization of bioenergy. In this work, we review the impact of fossil fuels burning, which is predominantly used to fulfill global energy demand, on the quality of the environment. We also assess the status of biofuel production and utilization and discuss its potential to clean the environment. The complications associated with biofuel manufacturing using photosynthetic microorganisms are discussed and directed evolution for targeted phenotypes and targeted delivery of nutrients are proposed as potential strategies to increase the biomass production.