Acute successful management of a chainsaw injury to the roof of the left chest.

This case report chronicles the experience of prompt resuscitation, beginning with the patient, and immediate surgery following the fatal malfunction of a chainsaw. The injuries were atypical for chainsaw injuries and involved total transection of the left subclavian artery and vein, total transection of the left brachial plexus and laceration to the apex of the left lung, among other injuries. A coordinated effort allowed for successful repair of the life- and limb-threatening injuries so that the patient could return to his young family in time for his 40th birthday.

Transcriptomic Signature of the Simulated Microgravity Response in Caenorhabditis elegans and Comparison to Spaceflight Experiments.

Given the growing interest in human exploration of space, it is crucial to identify the effects of space conditions on biological processes. Here, we analyze the transcriptomic response of Caenorhabditis elegans to simulated microgravity and observe the maintained transcriptomic response after returning to ground conditions for four, eight, and twelve days. We show that 75% of the simulated microgravity-induced changes on gene expression persist after returning to ground conditions for four days while most of these changes are reverted after twelve days. Our results from integrative RNA-seq and mass spectrometry analyses suggest that simulated microgravity affects longevity-regulating insulin/IGF-1 and sphingolipid signaling pathways. Finally, we identified 118 genes that are commonly differentially expressed in simulated microgravity- and space-exposed worms. Overall, this work provides insight into the effect of microgravity on biological systems during and after exposure.

Bioconjugation of Meldrum's acid activated furan: A detergent compatible assay for protein quantitation.

A simple yet efficient assay for the quantitation of proteins ranging from plasma proteins to purified proteins from whole cell lysate, based on the bioconjugation reaction between protein and Meldrum's acid Activated Furan (MAF) is described. This easy to use, sensitive method is based on the conjugation of amine functionalities present on the protein with MAF to form the corresponding Donor Acceptor Stenhouse Adducts (DASAs) with characteristic absorption in the visible region. The reaction is rapid as well as reproducible and shows a proportionate increase in color change over a broad range of protein concentration. The assay was found to be sensitive up to 0.125 mg/mL concentration of the protein and was compatible with most of the commonly employed detergents and isolation protocols which makes it ideal for the estimation of protein samples containing detergents. Another striking feature of this protocol is its tolerance towards other major interference contributors such as chelating agents, reducing agents, carbohydrates and protease inhibitors.

Drug infused Al2O3-bioactive glass coatings toward the cure of orthopedic infection.

A unique implant coated substrate with dual-drug-eluting system exhibiting antibacterial, anti-inflammatory, and bone regenerative capacity has been fabricated using spray pyrolysis deposition (SPD) method. Bioglass (BG) and BG-alumina (BG-Al) composites coatings with different concentrations of Al incorporated on BG network over the Cp-Ti substrate were fabricated using SPD technique. Phase purity of BG and BG-Al composites were analyzed by XRD in which Na2Ca2Si3O9 and ß-Na2Ca4(PO4)2SiO4) and Na7.15(Al7.2Si8.8O32) phases were formed. Surface morphology of the coated substrates was analyzed by SEM. Uniformity of the coatings were evaluated by surface profilometer and the uniform distribution the nanoparticles were confirmed with Elemental mapping. Systematically, each apatite layer formation on coated substrate was confirmed by immersing the samples for 1, 3, and 7 days in simulated body fluid and the needle-like structure was characterized using SEM. Cumulative release of Tetracycline hydrochloride (Tet) antibiotic and Dexamethasone (Dex) anti-inflammatory drug-loaded BG-Al and BG-Al composite-coated substrate were studied for 24 h. Antibacterial activity of the coated substrates were evaluated by time-dependent growth inhibition and minimal inhibitory concentration (MIC) assays in which BG-Al and BG-Al composite loaded with Tet showed considerable growth inhibition against S. aureus. Osteoblast-like cells (MG-63) exhibited profound proliferation with no cytotoxic effects which was due to release of Dex drug-coated substrates. Thus, surface modification of Cp-Ti substrate with BG, BG-Al composites coatings loaded with Tet and Dex drug can be considered for post-operative orthopedic implant infection application.

Burkholderia cepacia Bacteremia Complicated by Intracranial Abscesses and Immune Reconstitution Inflammatory Syndrome in a Renal Transplantation Recipient.

Burkholderia cepacia is almost always a colonizing organism rather than an infecting organism, but it may be pathogenic in immunocompromised individuals when isolated from body fluids that are ordinarily sterile. When recovered from blood culture it may present infection, pseuedo infection, or actual infection from contaminated intravenous fluids. We present a case of a renal transplant recipient patient who developed B. cepacia bacteremia following central venous cannulation. The subsequent clinical course was of worsening quadriparesis, which on neuroimaging revealed multiple brain and spinal abscesses. Following two weeks of intravenous antibiotics, his clinical features further worsened and the size of lesions further increased, which was suggestive of immune reconstitution inflammatory syndrome. With an increased steroid dose and continuation of the same anti-biotics, there was a regression of the lesions and significant clinical improvement.

In vitro anticancer activity of ethanol extract of Adhatoda vasica Nees on human ovarian cancer cell lines.

BACKGROUND: Ovarian cancer causes more deaths than any other cancer of the female reproductive system because there is no effective screening and most women are diagnosed at advanced stages. The probability of survival at 5 years is less than 30%, and the limitation is that it will not respond to chemotherapy protocol and surgery as well. Moreover, some evidence have shown potential anticancer properties of flavonoids, protective chemicals in plant foods, such as being an antioxidant, antiestrogenic, antiproliferative, and antiinflammatory. In this study, the anticancer activity of crude ethanol extracts of leaves from Adhatoda vasica was investigated. RESULTS: By the application of a cell-based assay, the LC 50 value of the A. vasica which showed anticancer effect was used for further studies. The cell line treated with LD 50 value of A. vasica extracts was observed for 0 h, 24 h, and 48 h to reveal the inhibition of the metastatic property in treated PA1 cells. The mRNA isolated from the teratocarcinoma PA1 cells treated with the A. vasica extract was further converted to cDNA and was amplified for the analysis of the p53 gene, p21 gene, and GAPDH gene expression. The expression in treated cells and the untreated control indicated the activity of the A. vasica extract against the ovarian cancer. CONCLUSION: The present study suggested the antiproliferative and antimetastatic effects of medicinal plant A. vasica on PA1 cells.

Bioactive, degradable and multi-functional three-dimensional membranous scaffolds of bioglass and alginate composites for tissue regenerative applications.

With a worldwide increase in the aged populace and associated geriatric diseases, there is an enormous need for the regeneration of degenerated organ systems. For this purpose, bioactive glass particulate (nBG) integrated alginate (Alg) composite membrane scaffolds were fabricated by a sol-gel assisted freeze-drying method and validated for their multifunctional utility in regenerative medicine. The presence of the combeite highly crystalline structure of nBG and Alg amorphous broad peaks were confirmed. Repetitive peaks from acids along with stretching confirmed the chemical interactions of the composites. Swelling ability, porosity, and in vitro degradation and biomineralization were analysed for up to 7 days. The results indicated that reduced swelling and degradation enhanced apatite formation. Hemocompatibility and the hemostatic properties on scaffolds were also systematically investigated. Additionally, significant cyto-compatibility and proliferation were noted in a culture with KB3-1. Further 3-D co-cultures with HDF cells and KB3-1 cells exhibited spheroid formation on Alg, nBG/Alg and nBG-Zr/Alg with profound dynamism required to establish organoids of interest. Thus, the results indicate that these 3D hydrogel membranes could offer infinite possibilities in the field of regenerative medicine, notably as an extracellular matrix (ECM) supporting the regeneration of bone, intra-vascularization, and neo-tissue formation, such as cartilage and ligaments.

Impact of copper on in-vitro biomineralization, drug release efficacy and antimicrobial properties of bioactive glasses.

This study highlights the incorporation of copper in the bioactive glasses (BAG) network that greatly influences the morphological, structural and biological properties. By increasing the copper incorporation in BAG, increment in cell volume was obtained from XRD patterns, and concomitantly, dominant phosphate bands and latent silica bands were observed by FT-IR and Raman spectroscopic results. The Cu addition also affected particle appearance to vary from spherical to cluster-like cubes in 1.5% and 2.5% copper-doped BAG. Due to the mesoporous network 1.5% and 2.5% copper-doped BAG showed enhanced release of anti-inflammatory drugs such as Acetaminophen (ACE) and Ibuprofen (IBU) in which, the drug release profiles showed best fit with kinetic models of First order, Korsmeyar-Peppas and Higuchi. Copper doping influences the lattice of BAG, as a result morphology and porosity varied, which regulates the ionic dissolution, hence, prompting bioactivity was perceived from 1.5% and 2.5% copper-doped bioactive glasses (Cu-BGs). Moreover, 2.5% Cu-BG and 1.5% Cu-BG showed highest rate of ROS detection, as well as improved antimicrobial activity. This study established that up to certain proportion of copper incorporation in BAG network, potentially enhances the biomineralization and turns the morphology towards minimal size with mesoporous nature. Due to the abundance in oral microbial exposure, copper amplifies the superior antimicrobial properties, and Cu-BGs act as a drug carrier to load ACE and IBU, which potentially up-regulate the healing properties in dental application.

Reference-Free Traction Force Microscopy Platform Fabricated via Two-Photon Laser Scanning Lithography Enables Facile Measurement of Cell-Generated Forces.

Cells sense and respond to the physical nature of their microenvironment by mechanically probing their surroundings via cytoskeletal contractions. The material response to these stresses can be measured via traction force microscopy (TFM). Traditional TFM platforms present several limitations including variable spatial resolution, difficulty in attaining the full three-dimensional (3D) deformation/stress profile, and the requirement to remove or relax the cells being measured to determine the zero-stress state. To overcome these limitations, we developed a two-photon, photochemical coupling approach to fabricate a new TFM platform that provides high-resolution control over the 3D placement of fluorescent fiducial markers for facile measurement of cell-generated shear and normal components of traction forces. The highly controlled placement of the 3D marker array provides a built-in, zero stress state eliminating the need to perturb the cells being measured while also providing increased throughput. Using this platform, we discovered that the magnitude of cell-generated shear and normal force components are linked both spatially and temporally. The facile nature and increased throughput of measuring cell-generated forces afforded by this new platform will be useful to the mechanotransduction community and others.

Insights Into the Mineralogy and Surface Chemistry of Extracellular Biogenic S0 Globules Produced by Chlorobaculum tepidum.

Elemental sulfur (S0) is produced and degraded by phylogenetically diverse groups of microorganisms. For Chlorobaculum tepidum, an anoxygenic phototroph, sulfide is oxidized to produce extracellular S0 globules, which can be further oxidized to sulfate. While some sulfur-oxidizing bacteria (e.g., Allochromatium vinosum) are also capable of growth on commercial S0 as an electron donor, C. tepidum is not. Even colloidal sulfur sols, which appear indistinguishable from biogenic globules, do not support the growth of C. tepidum. Here, we investigate the properties that make biogenic S0 globules distinct from abiotic forms of S0. We found that S0 globules produced by C. tepidum and abiotic S0 sols are quite similar in terms of mineralogy and material properties, but the two are distinguished primarily by the properties of their surfaces. C. tepidum's globules are enveloped by a layer of organics (protein and polysaccharides), which results in a surface that is fundamentally different from that of abiotic S0 sols. The organic coating on the globules appears to slow the aging and crystallization of amorphous sulfur, perhaps providing an extended window of time for microbes in the environment to access the more labile forms of sulfur as needed. Overall, our results suggest that the surface of biogenic S0 globules may be key to cell-sulfur interactions and the reactivity of biogenic S0 in the environment.

Single Nucleotide Polymorphism-Based Noninvasive Prenatal Testing: Experience in India.

INTRODUCTION: Noninvasive prenatal testing (NIPT) has revolutionized prenatal screening for chromosomal aneuploidies in some countries. Its implementation has been sporadic in developing countries. Given the genetic variation of the people in different countries, we evaluated the performance of the SNP-based NIPT in India . MATERIALS AND METHODS: The Panorama™ NIPT was performed in 516 pregnancies, which had tested intermediate-to-high risk on conventional first and second trimester screening. Results were confirmed either by invasive diagnostic testing or by clinical evaluation after birth. RESULTS: Of 511 samples analyzed, results were obtained in 499 (97.7%). Of these, 480 (98.2%) were low risk and 19 were high risk. A sensitivity of 100% was obtained for detection of trisomies 21, 18, 13 and sex chromosomal abnormalities. The specificity ranged from 99.3 to 100% for abnormalities tested. Taken together, the positive predictive value for trisomies 21, 18, 13 and monosomy X was 85.7%. The average fetal fraction was 8.2%, which is lower than the average observed elsewhere. CONCLUSION: This is the first report of detailed experience with NIPT in India and demonstrates comparable performance in all aspects of testing to the results elsewhere.

Effects of liquid cultivation on gene expression and phenotype of C. elegans.

BACKGROUND: Liquid cultures have been commonly used in space, toxicology, and pharmacology studies of Caenorhabditis elegans. However, the knowledge about transcriptomic alterations caused by liquid cultivation remains limited. Moreover, the impact of different genotypes in rapid adaptive responses to environmental changes (e.g., liquid cultivation) is often overlooked. Here, we report the transcriptomic and phenotypic responses of laboratory N2 and the wild-isolate AB1 strains after culturing P0 worms on agar plates, F1 in liquid cultures, and F2 back on agar plates. RESULTS: Significant variations were found in the gene expressions between the N2 and AB1 strains in response to liquid cultivation. The results demonstrated that 8-34% of the environmental change-induced transcriptional responses are transmitted to the subsequent generation. By categorizing the gene expressions for genotype, environment, and genotype-environment interactions, we identified that the genotype has a substantial impact on the adaptive responses. Functional analysis of the transcriptome showed correlation with phenotypical changes. For example, the N2 strain exhibited alterations in both phenotype and gene expressions for germline and cuticle in axenic liquid cultivation. We found transcript evidence to approximately 21% of the computationally predicted genes in C. elegans by exposing the worms to environmental changes. CONCLUSIONS: The presented study reveals substantial differences between N2 and AB1 strains for transcriptomic and phenotypical responses to rapid environmental changes. Our data can provide standard controls for future studies for the liquid cultivation of C. elegans and enable the discovery of condition-specific genes.

Adverse Immunologically Mediated Oral Mucosal Reactions to Systemic Medication: Lichenoid Tissue Reaction/Interface Dermatitis-Stomatitis, Autoimmune Vesiculobullous Disease, and IgE-Dependent and Immune Complex Reactions.

Drug-induced hypersensitivity immune reactions are exaggerated immunoinflammatory responses to allergenic components of the medications that occur in genetically susceptible subjects. The type of hypersensitivity immune response generated, whether antibody mediated or T cell mediated, or an immune complex reaction is determined by multiple factors, including the molecular characteristics of the allergen, the route of administration of the medication, the manner of presentation of the allergen by antigen-presenting cells to naïve T cells, the repertoire of the T cell receptors, and the cytokine profile within the microenvironment. This review deals with the clinical and histopathological aspects of adverse immunologically mediated oral mucosal reactions to systemic medication. We elaborate on diseases showing features of lichenoid tissue reaction/interface dermatitis-stomatitis, autoimmune vesiculobullous oral lesions, and immunoglobulin E- (IgE-) and immune complex-mediated oral reactions to drugs.

Microstructured Elastomer-PEG Hydrogels via Kinetic Capture of Aqueous Liquid-Liquid Phase Separation.

Heterogeneous hydrogels with desired matrix complexity are studied for a variety of biomimetic materials. Despite the range of such microstructured materials described, few methods permit independent control over microstructure and microscale mechanics by precisely controlled, single-step processing methods. Here, a phototriggered crosslinking methodology that traps microstructures in liquid-liquid phase-separated solutions of a highly elastomeric resilin-like polypeptide (RLP) and poly(ethylene glycol) (PEG) is reported. RLP-rich domains of various diameters can be trapped in a PEG continuous phase, with the kinetics of domain maturation dependent on the degree of acrylation. The chemical composition of both hydrogel phases over time is assessed via in situ hyperspectral coherent Raman microscopy, with equilibrium concentrations consistent with the compositions derived from NMR-measured coexistence curves. Atomic force microscopy reveals that the local mechanical properties of the two phases evolve over time, even as the bulk modulus of the material remains constant, showing that the strategy permits control of mechanical properties on micrometer length scales, of relevance in generating mechanically robust materials for a range of applications. As one example, the successful encapsulation, localization, and survival of primary cells are demonstrated and suggest the potential application of phase-separated RLP-PEG hydrogels in regenerative medicine applications.

Distribution and Diversity of Rhodopsin-Producing Microbes in the Chesapeake Bay.

Although sunlight is an abundant source of energy in surface environments, less than 0.5% of the available photons are captured by (bacterio)chlorophyll-dependent photosynthesis in plants and bacteria. Metagenomic data indicate that 30 to 60% of the bacterial genomes in some environments encode rhodopsins, retinal-based photosystems found in heterotrophs, suggesting that sunlight may provide energy for more life than previously suspected. However, quantitative data on the number of cells that produce rhodopsins in environmental systems are limited. Here, we use total internal reflection fluorescence microscopy to show that the number of free-living microbes that produce rhodopsins increases along the salinity gradient in the Chesapeake Bay. We correlate this functional data with environmental data to show that rhodopsin abundance is positively correlated with salinity and with indicators of active heterotrophy during the day. Metagenomic and metatranscriptomic data suggest that the microbial rhodopsins in the low-salinity samples are primarily found in Actinobacteria and Bacteroidetes, while those in the high-salinity samples are associated with SAR-11 type AlphaproteobacteriaIMPORTANCE Microbial rhodopsins are common light-activated ion pumps in heterotrophs, and previous work has proposed that heterotrophic microbes use them to conserve energy when organic carbon is limiting. If this hypothesis is correct, rhodopsin-producing cells should be most abundant where nutrients are most limited. Our results indicate that in the Chesapeake Bay, rhodopsin gene abundance is correlated with salinity, and functional rhodopsin production is correlated with nitrate, bacterial production, and chlorophyll a We propose that in this environment, where carbon and nitrogen are likely not limiting, heterotrophs do not need to use rhodopsins to supplement ATP synthesis. Rather, the light-generated proton motive force in nutrient-rich environments could be used to power energy-dependent membrane-associated processes, such as active transport of organic carbon and cofactors, enabling these organisms to more efficiently utilize exudates from primary producers.

Aneurysmal hemorrhage in a pregnant patient with coarctation of aorta: An anesthetic challenge.

A 25years old female patient with pregnancy of 16weeks (G2 P1), diagnosed to have distal anterior cerebral artery aneurysm (DACA) with Hunt & Hess grade I, subarachnoid hemorrhage (SAH) and coexisting atretic type of aortic coarctation posted for aneurysmal clipping under general anesthesia is a challenge to anesthesiologists in perioperative period. Hypertensive surges in a pregnant patient may result in rupture of aneurysms. Mortality in the mothers with CoA has been reported to be in the range of 0 to 9%. Anesthetic management of a pregnancy with CoA and SAH has never been reported.

Vascular Cells in Blood Vessel Wall Development and Disease.

The vessel wall is composed of distinct cellular layers, yet communication among individual cells within and between layers results in a dynamic and versatile structure. The morphogenesis of the normal vascular wall involves a highly regulated process of cell proliferation, migration, and differentiation. The use of modern developmental biological and genetic approaches has markedly enriched our understanding of the molecular and cellular mechanisms underlying these developmental events. Additionally, the application of similar approaches to study diverse vascular diseases has resulted in paradigm-shifting insights into pathogenesis. Further investigations into the biology of vascular cells in development and disease promise to have major ramifications on therapeutic strategies to combat pathologies of the vasculature.

Aqueous Liquid-Liquid Phase Separation of Resilin-Like Polypeptide/Polyethylene Glycol Solutions for the Formation of Microstructured Hydrogels.

Multiple approaches to generate microstructured hydrogels have emerged in order to control microscale properties for applications ranging from mechanical reinforcement to regenerative medicine. Here, we report new heterogeneous hybrid hydrogels comprising emerging resilin-like polypeptides (RLPs); the hydrogels can be engineered with controlled microstructure and distinct micromechanical properties via the liquid-liquid phase separation (LLPS) of aqueous solutions of the RLPs and poly(ethylene glycol) (PEG). The microstructure in the hydrogels was captured by cross-linking a phase-separated RLP and PEG solution via a Mannich-type reaction with the cross-linker tris(hydroxymethyl phosphine) (THP). Phase diagrams of the RLP/PEG system were generated in order to define solution parameters that would yield micron-scale domains in the hydrogels with diameters on the order of 20-90 µm; the production of RLP- and PEG-rich domains with these dimensions was confirmed via confocal microscopy. The hydrogel mechanical properties were assessed via oscillatory rheology and atomic force microscopy (AFM), with the hydrogels exhibiting a moderate bulk shear storage modulus (ca. 600 Pa) and micromechanical properties of the domains (Young's modulus ca. 13 kPa) that were distinct from those of the matrix (ca. 6 kPa). These results demonstrate that tuning the parameters of the aqueous-aqueous phase-separated RLP/PEG solutions provides a simple, straightforward methodology for fabricating microstructured protein-containing hydrogels, without extensive material processing or purification. Given the unusual mechanical properties of the resilins, these methods potentially could be useful for engineering the micromechanical properties and cellular behavior in phase-separated protein-polymer hydrogels.

HIV-Associated Oral Mucosal Melanin Hyperpigmentation: A Clinical Study in a South African Population Sample.

Objective. The aim of the study was to determine the prevalence of HIV-associated oral mucosal melanin hyperpigmentation (HIV-OMH) in a specific population of HIV-seropositive South Africans and to analyse the associations between HIV-OMH clinical features and the demographic and immunological characteristics of the study cohort. Material and Methods. This cross-sectional study included 200 HIV-seropositive Black subjects. The collected data comprised age, gender, CD4+ T cell count, viral load, systemic disease, medications, oral site affected by HIV-OMH, extent (localized or generalized), intensity of the pigmentation (dark or light), and smoking and snuff use. Results. Overall, 18.5% of the study cohort had HIV-OMH. Twenty-two and a half percent had OMH that could not with confidence be attributed to HIV infection, and 59% did not have any OMH. There was a significant but weak association between smoking and the presence of HIV-OMH. Conclusions. The prevalence of HIV-OMH in the study population was 18.5%, the gingiva being the most commonly affected site. It appears that the CD4+ T cell count does not play any role in the biopathology of HIV-OMH.

Extracting Diffusive States of Rho GTPase in Live Cells: Towards In Vivo Biochemistry.

Resolving distinct biochemical interaction states when analyzing the trajectories of diffusing proteins in live cells on an individual basis remains challenging because of the limited statistics provided by the relatively short trajectories available experimentally. Here, we introduce a novel, machine-learning based classification methodology, which we call perturbation expectation-maximization (pEM), that simultaneously analyzes a population of protein trajectories to uncover the system of diffusive behaviors which collectively result from distinct biochemical interactions. We validate the performance of pEM in silico and demonstrate that pEM is capable of uncovering the proper number of underlying diffusive states with an accurate characterization of their diffusion properties. We then apply pEM to experimental protein trajectories of Rho GTPases, an integral regulator of cytoskeletal dynamics and cellular homeostasis, in vivo via single particle tracking photo-activated localization microscopy. Remarkably, pEM uncovers 6 distinct diffusive states conserved across various Rho GTPase family members. The variability across family members in the propensities for each diffusive state reveals non-redundant roles in the activation states of RhoA and RhoC. In a resting cell, our results support a model where RhoA is constantly cycling between activation states, with an imbalance of rates favoring an inactive state. RhoC, on the other hand, remains predominantly inactive.