Translational optical coherence elastography for assessment of systemic sclerosis

Systemic sclerosis (SSc‐scleroderma) is an autoimmune disorder with high mortality rate that results in excessive accumulation of collagen in the skin and internal organs. Currently, the modified Rodnan Skin Score (mRSS) is the gold standard for evaluating the dermal thickening due to SSc. However, mRSS has noticeable inter‐ and intra‐observer variabilities as quantified by the interclass correlation coefficient (ICC: 0.6‐0.75). In this work, optical coherence elastography (OCE) combined with structural optical coherence tomography (OCT) image analysis was used to assess skin thickness in 12 SSc patients and healthy volunteers. Inter‐ (ICC: 0.62‐0.99) and intra‐observer (ICC > 0.90) assessment of OCT/OCE showed excellent reliability. Clinical assessments, including histologically assessed dermal thickness (DT), mRSS, and site‐specific mRSS (SMRSS) were also performed for further validation. The OCE and OCT results from the forearm demonstrated the highest correlation (OCE: 0.78, OCT: 0.65) with SMRSS. Importantly, OCE and OCT had stronger correlations with the histological DT (OCT: r = .78 and OCE: r = .74) than SMRSS (r = .57), indicating the OCT/OCE could outperform semi‐quantitative clinical assessments such as SMRSS. Overall, these results demonstrate that OCT/OCE could be useful for rapid, noninvasive and objective assessments of SSc onset and monitoring skin disease progression and treatment response.     

Radiation dose measurements in a dental orthopantogram unit using indigenously developed optically stimulated luminescence dosimeters

Dental orthopantogram (OPG)/cone beam computed tomography (CBCT) scanners are gaining popularity due to their 3D imaging with multiplanar view that provides clinical benefits over conventional dental radiography systems. Dental OPG/CBCT provides optimal visualization of adjacent overlaying anatomical structures that will be superpositioned in any single projection. The characteristics of indigenously developed optically stimulated luminescence dosimeters, namely, aluminium oxide doped with carbon (Al₂O₃:C), lithium magnesium phosphate doped with terbium and boron (LiMgPO₄:Tb,B) and lithium calcium aluminium fluoride doped with europium and yttrium (LiCaAlF₆:Eu,Y) were evaluated for their use in dental dosimetry. The dose?response of these dosimeters was studied at X‐ray energies 60 kV, 70 kV and 81 kV. Radiation doses were also measured using Gafchromic film for comparison. Radiation dose was measured at eight different locations of a polymethyl methacrylate (PMMA) head phantom including eyes. The optically stimulated luminescence (OSL) sensitivity of LiMgPO₄:Tb,B is about 1.5 times and LiCaAlF₆:Eu, is about 20 times higher than the sensitivity of Al₂O₃:C. It was found that measured radiation doses by the three optically stimulated luminescence dosimeters (OSLDs) and Gafchromic film in the occipital region (back side) of a PMMA phantom, were consistent but variations in dose at other locations were significantly higher. The three OSLDs used in this study were found to be suitable for radiation dose measurement in dental units.     

Grades of undernutrition and socioeconomic status influence cognitive development in school children of Kolkata

Cognitive development of children is influenced by different environmental factors like nutritional and socio‐economic status. The objectives of the present study were to determine the influence of grades of undernutrition and socio‐economic status (SES) on the cognitive development of school children of Kolkata. Five hundred sixty six (566) school children having 5–12 years of age were selected from different schools of Kolkata. The cognitive development was measured by the scores of Raven's colored progressive matrices (RCPM). The chronic and acute nutritional statuses were measured from height‐for‐age (HAZ) and weight‐for‐age (WAZ) Z scores respectively with reference to the values of WHO. SES was determined by updated Kuppuswamy scale. The prevalences of undernutrition in the observed children were 57.95% (according to HAZ) and 52.82% (according to WAZ). The age dependent growth curve of RCPM scores of the observed children remains in between the 10th and 25th centile of British children. The children belonging to superior and intellectual deficit IQ classes were 21.55 and 36.40%, respectively of the total subjects. Most of the subjects belong to lower middle (39.93%) and upper middle (36.40%) class of SES. RCPM scores of school children were gradually decreased with the grades of undernutrition and SES. RCPM scores were significantly correlated with HAZ, WAZ, SES, age, and sex (P < 0.001) and strongly associated with HAZ, SES, age, and sex (P < 0.001, P < 0.05). Present study indicates that cognitive development of school children of Kolkata is influenced by the grade of undernutrition and SES. Am J Phys Anthropol 156:274–285, 2015. © 2014 Wiley Periodicals, Inc.     

Environmental Factors Associated with High Fly Densities and Diarrhea in Vellore,India

Diarrhea causes significant morbidity and mortality in Indian children under 5 years of age. Flies carry enteric pathogens and may mediate foodborne infections. In this study, we characterized fly densities as a determinant of infectious diarrhea in a longitudinal cohort of 160 urban and 80 rural households with 1,274 individuals (27% under 5 years of age) in Vellore, India. Household questionnaires on living conditions were completed at enrollment. Fly abundance was measured during the wet and dry seasons using fly ribbons placed in kitchens. PCRs for enteric bacteria, viruses, and protozoa were performed on 60 fly samples. Forty-three (72%) fly samples were positive for the following pathogens: norovirus (50%), Salmonella spp. (46.7%), rotavirus (6.7%), and Escherichia coli (6.7%). Ninety-one episodes of diarrhea occurred (89% in children under 5 years of age). Stool pathogens isolated in 24 of 77 (31%) samples included E. coli, Shigella spp., Vibrio spp., Giardia, Cryptosporidium, and rotavirus. Multivariate log-linear models were used to explore the relationships between diarrhea and fly densities, controlling for demographics, hygiene, and human-animal interactions. Fly abundance was 6 times higher in rural than urban sites (P < 0.0001). Disposal of garbage close to homes and rural living were significant risk factors for high fly densities. The presence of latrines was protective against high fly densities and diarrhea. The adjusted relative risks of diarrheal episodes and duration of diarrhea, associated with fly density at the 75th percentile, were 1.18 (95% confidence interval [CI], 1.03 to 1.34) and 1.15 (95% CI, 1.02 to 1.29), respectively. Flies harbored enteric pathogens, including norovirus, a poorly documented pathogen on flies.     

Computational prediction of small molecules with predicted binding to FGFR3 and testing biological effects in bone cells

Activating anabolic receptor-mediated signaling is essential for stimulating new bone formation and for promoting bone healing in humans. Fibroblast growth factor receptor (FGFR) 3 is reported to be an important positive regulator of osteogenesis. Presently, recombinant proteins are used to stimulate FGFR3 function but have limitations for therapy due to expense and stability. Therefore, there is a need for identification of novel small molecules binding to FGFR3 that promote biological function. In silico molecular docking and high-throughput virtual screening on zinc database identified seven compounds predicted to bind to an active site within the βCʹ-βE loop, specific to FGFR3. All seven compounds fall within an acceptable range of ADME/T properties. Four compounds showed a 30–65% oral absorption rate. Density functional theory analysis revealed a high HOMO-LUMO gap, reflecting high molecular stability for compounds 14977614 and 13509082. Five compounds exhibited mutagenicity, while the other three compounds presented irritability. Computational mutagenesis predicted that mutating G322 affected compound binding to FGFR3. Molecular dynamics simulation revealed compound 14977614 is stable in binding to FGFR3. Furthermore, compound 14977614, with an oral absorption rate of 60% and high molecular stability, produced significant increases in both proliferation and differentiation of bone marrow stromal cells in vitro. Anti-FGFR3 treatment completely blocked the stimulatory effect of 14977614 on BMSC proliferation. Ex vivo treatment of mouse calvaria in organ culture for seven days with 14977614 increased mineralization and expression levels of bone formation markers. In conclusion, computational analyses identified seven compounds that bind to the FGFR3, and in vitro studies showed that compound 14977614 exerts significant biological effects on osteogenic cells.     

Novel Synthetic Biscoumarins Target Tumor Necrosis Factor-α in Hepatocellular Carcinoma in Vitro and in Vivo

TNF is a pleotropic cytokine known to be involved in the progression of several pro-inflammatory disorders. Many therapeutic agents have been designed to counteract the effect of TNF in rheumatoid arthritis as well as a number of cancers. In the present study we have synthesized and evaluated the anti-cancer activity of novel biscoumarins in vitro and in vivo. Among new compounds, BIHC was found to be the most cytotoxic agent against the HepG2 cell line while exhibiting less toxicity toward normal hepatocytes. Furthermore, BIHC inhibited the proliferation of various hepatocellular carcinoma (HCC) cells in a dose- and time-dependent manner. Subsequently, using in silico target prediction, BIHC was predicted as a TNF blocker. Experimental validation was able to confirm this hypothesis, where BIHC could significantly inhibit the recombinant mouse TNF-α binding to its antibody with an IC₅₀ of 16.5 μm. Furthermore, in silico docking suggested a binding mode of BIHC similar to a ligand known to disrupt the native, trimeric structure of TNF, and also validated with molecular dynamics simulations. Moreover, we have demonstrated the down-regulation of p65 phosphorylation and other NF-κB-regulated gene products upon BIHC treatment, and on the phenotypic level the compound shows inhibition of CXCL12-induced invasion of HepG2 cells. Also, we demonstrate that BIHC inhibits infiltration of macrophages to the peritoneal cavity and suppresses the activity of TNF-α in vivo in mice primed with thioglycollate broth and lipopolysaccharide. We comprehensively validated the TNF-α inhibitory efficacy of BIHC in an inflammatory bowel disease mice model.     

Light Intensity Alters the Extent of Arsenic Toxicity in Helianthus annuus L. Seedlings

The present study is aimed at assessing the extent of arsenic (As) toxicity under three different light intensities—optimum (400 μmole photon m^?2 s^?1), sub-optimum (225 μmole photon m^?2 s^?1), and low (75 μmole photon m^?2 s^?1)—exposed to Helianthus annuus L. var. DRSF-113 seedlings by examining various physiological and biochemical parameters. Irrespective of the light intensities under which H. annuus L. seedlings were grown, there was an As dose (low, i.e., 6 mg kg^?1 soil, As₁; and high, i.e., 12 mg kg^?1 soil, As₂)-dependent decrease in all the growth parameters, viz., fresh mass, shoot length, and root length. Optimum light-grown seedlings exhibited better growth performance than the sub-optimum and low light-grown seedlings; however, low light-grown plants had maximum root and shoot lengths. Accumulation of As in the plant tissues depended upon its concentration used, proximity of the plant tissue, and intensity of the light. Greater intensity of light allowed greater assimilation of photosynthates accompanied by more uptake of nutrients along with As from the medium. The levels of chlorophyll a, b, and carotenoids declined with increasing concentrations of As. Seedlings acquired maximum Chl a and b under optimum light which were more compatible to face As₁ and As₂ doses of As, also evident from the overall status of enzymatic (SOD, POD, CAT, and GST) and non-enzymatic antioxidant (Pro).     

Some findings of FADD knockdown in inhibition of HIV-1 replication in Jurkat cells and PBMCs

Fas-associated protein with death domain (FADD) is a key adaptor molecule transmitting the death signal mediated by death receptors, and it is also required for T cell proliferation. A recent study indicated that FADD is able to affect HIV-1 production, but the mechanism is not known. Using the susceptible Jurkat cell line and peripheral blood mononuclear cells, we studied the effects of FADD on HIV-1 production. TaqMan RT-PCR was used to quantify HIV-1 viral RNA copies, and Western blot analysis was used to detect protein expression. FADD knockdown decreased HIV-1 replication and inactivated caspase-3 activity in the cells and blocked CD4 translocation to the lipid rafts of the plasma membrane. Reduced expression of FADD suppressed TCR signaling through downregulation of TCR, CD3, and Zap-70 in response to HIV-1 infection and blocked the trafficking of TCR, CD3, CD28, and Zap-70 to lipid rafts, leading to reduced activation of NF-κB and NFAT, which are required for HIV-1 replication. FADD knockdown diminished caspase-8 migration to lipid rafts and its expression in response to HIV-1 infection. These results indicate that FADD, as a host pro-apoptotic protein, plays important roles in regulating HIV-1 replication and production in several ways, and apoptotic pathway inhibition is able to decrease HIV-1 replication and production.     

Utility of indels for species-level identification of a biologically complex plant group: a study with intergenic spacer in Citrus

The Consortium of Barcode of Life plant working group proposed to use the defined portion of plastid genes rbcL and matK either singly or in combination as the standard DNA barcode for plants. But DNA barcode based identification of biologically complex plant groups are always a challenging task due to the occurrence of natural hybridization. Here, we examined the use of indels polymorphism in trnH-psbA and trnL-trnF sequences for rapid species identification of citrus. DNA from young leaves of selected citrus species were isolated and matK gene (~800 bp) and trnH-psbA spacer (~450 bp) of Chloroplast DNA was amplified for species level identification. The sequences within the group taxa of Citrus were aligned using the ClustalX program. With few obvious misalignments were corrected manually using the similarity criterion. We identified a 54 bp inverted repeat or palindrome sequence (27–80 regions) and 6 multi residues indel coding regions. Large inverted repeats in cpDNA provided authentication at the higher taxonomic levels. These diagnostics indel marker from trnH-psbA were successful in identifying different species (5 out of 7) within the studied Citrus except Citrus limon and Citrus medica. These two closely related species are distinguished through the 6 bp deletion in trnL-trnF. This study demonstrated that the indel polymorphism based approach easily characterizes the Citrus species and the same may be applied in other complex groups. Likewise other indels occurring intergenic spacer of chloroplast regions may be tested for rapid identification of other secondary citrus species.