In-vitro Wound Healing Assays: A Comprehensive Review.

Chronic non-healing wounds offer a tough clinical challenge for plastic and reconstructive surgeons. A systematic comprehension of the underlying mechanism of wound healing is essential for improved patient care. It is also required to know the complex and continuing armamentarium of therapeutic choices for treatment of chronic wounds. Wound assays are excellent way to apprehend the wound healing progression and the efficacy of various products that contribute and promote in wound healing. In the production sequence of wound care products also, human trials are often preceded by more economical in-vitro or preclinical studies. Wound healing assays are to understand the principal molecular mechanisms involved in repair progression and used in the exploration of impending therapeutics and designing treatment protocols intended for better curing. Several prototypes of wound healing have been established and are well-accepted. This review focuses on in vitro assays, as they are pre-requisite preliminary assays that are rapid, economical and ethical substitute to other complex assay models. Literature exploration for data sources were carried out through a meticulous search in indexed literature and various website centered educational research documents. In total 76 studies were earmarked from 286 most relevant scholarly articles. An extensive criterion to describe both in-vitro assays and in-vitro assays in wound healing has been used for probing the comments. Also, personal resources have been used for information gathering. The present manuscript provides a broad overview of various in-vitro assays with descriptive account of injury techniques with assessment of pros and cons of each. It also emphasized on three-dimensional culture assays, which utilizes bioengineered prototypes to exhibit various connections and the mechanisms of diverse sort of cells in the wound healing.

Evidence of a Hardening in the Cosmic Ray Proton Spectrum at around 166 TeV Observed by the GRAPES-3 Experiment.

We present the measurement of the cosmic ray proton spectrum from 50 TeV to 1.3 PeV using 7.81×10^{6} extensive air shower events recorded by the ground-based GRAPES-3 experiment between 1 January 2014 and 26 October 2015 with a live time of 460 day. Our measurements provide an overlap with direct observations by satellite and balloon-based experiments. The electromagnetic and muon components in the shower were measured by a dense array of plastic scintillator detectors and a tracking muon telescope, respectively. The relative composition of the proton primary from the air shower data containing all primary particles was extracted using the multiplicity distribution of muons which is a sensitive observable for mass composition. The observed proton spectrum suggests a spectral hardening at ∼166 TeV and disfavors a single power law description of the spectrum up to the Knee energy (∼3 PeV).

Reliability of the electric pulp tester as an indicator of pulpal anesthesia in primary maxillary posterior teeth to assess the need for supplemental injections: An observational clinical study.

Background: The primary maxillary molars occasionally remain sensitive during operative procedures even post the buccal supraperiosteal injection. This could be due to the widely flared palatal roots receiving accessory innervation from the palatal nerves. Identifying inadequate anesthesia upfront using the electric pulp test (EPT) would give vital information to the clinician on the need of a supplemental palatal injection. Aim: The aim of this study was to assess and evaluate the reliability of the EPT as an indicator of pulpal anesthesia in primary maxillary molars. Methodology: Fifty one primary maxillary molars were subjected to the EPT following a buccal supraperiosteal injection. During the operative procedure, the " Face Legs Activity Cry Consolability" (FLACC) scores were recorded. The outcome of the EPT was correlated with the results of the FLACC score using Pearson's Chi-square test.Results: The EPT results were correlated to the FLACC scores. Five out of the 10 primary maxillary second molars which responded to the EPT scored 0 on the FLACC scale. The remaining 5 teeth scored 1 on the FLACC scale. The P value was 0.056 which was not statistically significant. This infers that the EPT is not a reliable tool to assess the adequacy of pulpal anesthesia in primary maxillary second molars. Conclusion: From the results of the present study, it can be concluded that the EPT is not a reliable tool to be used as an indicator of pulpal anesthesia in primary maxillary molars.

Integrated assessment of drought vulnerability for water resources management of Bina basin in Central India.

Drought is an extreme event and its frequency is expected to increase in future under the imminent threats of climate change. The areas vulnerable to drought are increasing due to increase in the spatial extent and severity of droughts. This necessitates the need for development of an integrated framework for assessment of drought vulnerability, which will be vital for water resources management policies focused towards such vulnerable areas. An integrated drought vulnerability assessment framework has been developed considering the physical indicators that vary spatially, social indicators that vary spatially but their temporal variation may be at longer time-frames, and spatio-temporal drought indicators that vary spatially and temporally during various months during drought years. This framework has been tested for Bina basin located in the drought prone Bundelkhand region of Madhya Pradesh. The drought indicators used in the study include (i) Standardized Precipitation Index (SPI) for evaluating meteorological drought characteristics, (ii) Surface water Drought Index (SDI) for evaluating streamflow drought characteristics, and (iii) Groundwater Drought Index (GDI) for evaluating groundwater drought characteristics. Groundwater levels are being observed at quarterly (3 monthly) time step only. So the relationships between GDI and 3-m SPI, 6-m SPI, and 12-m SPI have been investigated. Based on the best correlation, the 12-m SPI can be used to represent the groundwater drought in Bina basin and has therefore been used to assess the monthly variability in the groundwater drought characteristics. The spatially varying physical indicators including basin reach (elevation band), land use pattern and soil type; the spatio-temporal drought indicators including soil moisture drought, surface water drought and groundwater drought, rainfall departure and number of consecutive dry days; and the spatially varying social indicators including infants and young children, illiterate population, marginal workers and rural population have been used for the development of a Drought Vulnerability Index (DVI). The integrated drought vulnerability assessment framework has been conceptualized on the basis of DVI. Four vulnerability classes have been defined and the study area falls in mild to moderate vulnerable class, based on the analysis carried out for the various drought years in the basin. Appropriate drought management plans and mitigation strategies need to be developed to target these vulnerable areas in Bina basin.

Combined application of ascorbic acid and endophytic N-fixing Azotobacter chroococcum Avi2 modulates photosynthetic efficacy, antioxidants and growth-promotion in rice under moisture deficit stress.

This group has previously reported the role of ascorbic acid (AA) as an antioxidant for survivability and ability to enhancing diazotrophic efficacy in Azotobacter chroococcum Avi2 under hydrogen peroxide (H2O2) stress. However, the present study showed the combined application of AA and Avi2 in drought-susceptible (IR64 and Naveen) and drought-tolerant (Ankit and Satyabhama) rice cultivars to determine their photosynthetic efficacy (chlorophyll fluorescence-imaging), antioxidants, and plant growth-promotion (PGP) under moisture deficit stress (MS, -60 kPa). The results indicated that combined application of AA and Avi2 significantly (p < 0.05) increased the total chlorophyll, relative water content, electrolytic leakage, super oxide dismutase, and catalase activities in all rice cultivars as compared to other MS treatments, whereas stress indicators like proline and H2O2 contents were proportionally increased under MS and their concentration were normalized under combined application of AA and Avi2. Photochemical quenching, non-photochemical quenching, photosynthetic electron transport rate, and the effective quantum efficiency were found to be increased significantly (p < 0.05) in Avi2 + AA as compared to other MS treatments. Moreover, rice roots harbored significantly (p < 0.05) higher copy number of nifH gene in Avi2 + AA treatment followed by Avi2 compared to flooded control and other MS treatments. Combined application of AA and Avi2 also increased the grain yield significantly (p < 0.05) by 7.09 % and 3.92 % in drought-tolerant (Ankit and Satyabhama, respectively) and 31.70 % and 34.19 % in drought-susceptible (IR64 and Naveen, respectively) rice cultivars compared to MS treatment. Overall, the present study indicated that AA along with Avi2 could be an effective formulation to alleviate MS vis à vis enhances PGP traits in rice.

Temporal mechanisms of myogenic specification in human induced pluripotent stem cells.

Understanding the mechanisms of myogenesis in human induced pluripotent stem cells (hiPSCs) is a prerequisite to achieving patient-specific therapy for diseases of skeletal muscle. hiPSCs of different origin show distinctive kinetics and ability to differentiate into myocytes. To address the unique cellular and temporal context of hiPSC differentiation, we perform a longitudinal comparison of the transcriptomic profiles of three hiPSC lines that display differential myogenic specification, one robust and two blunted. We detail temporal differences in mechanisms that lead to robust myogenic specification. We show gene expression signatures of putative cell subpopulations and extracellular matrix components that may support myogenesis. Furthermore, we show that targeted knockdown of ZIC3 at the outset of differentiation leads to improved myogenic specification in blunted hiPSC lines. Our study suggests that ß-catenin transcriptional cofactors mediate cross-talk between multiple cellular processes and exogenous cues to facilitate specification of hiPSCs to mesoderm lineage, leading to robust myogenesis.

Ascorbic acid formulation for survivability and diazotrophic efficacy of Azotobacter chroococcum Avi2 (MCC 3432) under hydrogen peroxide stress and its role in plant-growth promotion in rice (Oryza sativa L.).

Oxidative stress generates reactive oxygen species which causes cell damage of living organisms and are normally detoxified by antioxidants. Indirect reports signify the damages caused by reactive oxygen species and neutralized by antioxidant, but the direct evidence to confirm this hypothesis is still unclear. To validate our hypothesis, an attempt was made in a diazotrophic bacterium (Azotobacter chroococcum Avi2) as a biological system, and hydrogen peroxide (H2O2) and ascorbic acid were used as oxidative stress and antioxidant supplement, respectively. Additionally, rice plant-growth attributes by Avi2 was also assessed under H2O2 and ascorbic acid. Results indicated that higher concentration of H2O2 (2.5 mM-4.5 mM) showed the complete mortality of Avi2, whereas one ppm ascorbic acid neutralized the effect of H2O2. Turbidity, colony forming unit, DNA quantity, nifH gene abundance, indole acetic acid and ammonia productions were significantly (p < 0.5) increased by 11.93%, 17.29%, 19.80%, 74.77%, 71.89%, and 42.53%, respectively in Avi2-treated with 1.5 mM H2O2 plus ascorbic acid compared to 1.5 mM H2O2 alone. Superoxide dismutase was significantly (p < 0.5) increased by 60.85%, whereas catalase and ascorbate peroxidase activities were significantly (p < 0.05) decreased by 64.28% and 68.88% in Avi2-treated with 1.5 mM H2O2 plus ascorbic acid compared to 1.5 mM H2O2 alone. Germination percentage of three rice cultivars (FR13a, Naveen and Sahbhagi dhan) were significantly (p < 0.5) increased by 20%, 13.33%, and 4%, respectively in Avi2-treated with 0.6 mM H2O2 plus ascorbic acid compared with uninoculated control. Overall, this study indicated that ascorbic acid formulation neutralizes the H2O2-oxidative stress and enhances the survivability and plant growth-promoting efficacy of A. chroococcum Avi2 and therefore, it may be used as an effective formulation of bio-inoculants in rice under oxidative stress.

Measurement of the Electrical Properties of a Thundercloud Through Muon Imaging by the GRAPES-3 Experiment.

The GRAPES-3 muon telescope located in Ooty, India records rapid (∼10 min) variations in the muon intensity during major thunderstorms. Out of a total of 184 thunderstorms recorded during the interval of April 2011-December 2014, the one on December 1, 2014 produced a massive potential of 1.3 GV. The electric field measured by four well-separated (up to 6 km) monitors on the ground was used to help estimate some of the properties of this thundercloud, including its altitude and area that were found to be 11.4 km above mean sea level and ≥380 km^{2}, respectively. A charging time of 6 min to reach 1.3 GV implied the delivery of a power of ≥2 GW by this thundercloud that was moving at a speed of ∼60 km h^{-1}. This work possibly provides the first direct evidence for the generation of gigavolt potentials in thunderclouds that could also possibly explain the production of highest-energy (100 MeV) gamma rays in the terrestrial gamma-ray flashes.

Comparison of Patient Comfort During the First Stage of Conventional Versus Modified Two-Stage Inferior Alveolar Nerve Blocks in Pediatric Patients.

Our aim was to compare the comfort of pediatric patients during the first stage of the conventional 2-stage inferior alveolar nerve block (IANB) versus the modified 2-stage IANB. This was a parallel group, single-blinded, randomized controlled trial. Pediatric patients meeting the inclusion criteria were randomly allocated to 1 of the 2 groups receiving IANB, group 1 (31 subjects) utilized a conventional 2-stage technique, and group 2 (33 subjects) utilized a modified 2-stage approach. Patient comfort was assessed indirectly by 2 observers using the Face, Legs, Activity, Cry, Consolability (FLACC) scale during the first stage of the injections. FLACC score data between the 2 groups were analyzed using the chi-square test, and interobserver reliability was measured using kappa statistics. During the first stage of the injections, lower FLACC scores of 0 indicating a relaxed/comfortable patient experience were found in 57.6% of group 2 subjects compared with only 25.8% group 1 subjects, which was a statistically significant difference (p-value .013). Interobserver reliability assessed using kappa coefficients (κ = 0.812) indicated excellent agreement. FLACC scores for the pediatric patients during the first stage of the modified 2-stage IANB technique were significantly lower than during the first stage of the conventional 2-stage IANB technique.

Osteonecrosis of the Jaw among Patients Receiving Antiresorptive Medication: A 4-year Retrospective Study at a Tertiary Cancer Center, Kerala, India.

PURPOSE: Osteonecrosis of the jaw (ONJ) is a rare but complicated side effect of antiresorptive medications. The aim of the study is to evaluate the dental and drug-related factors related to ONJ among patients on these drugs at a tertiary cancer center, India. METHODOLOGY: A retrospective record review of patients who received antiresorptive medication at our center from 2011 to 2014 was done. The demographic factors, type, dosage, and duration of the medication and dental history were collected, and the data were entered an analyzed using Epidata software. RESULTS: A higher incidence of ONJ (8.1%) was noted in our sample (n = 183). Dental intervention after zoledronic acid (ZA) administration showed a statistical significance (P < 0.001). No significance (P value) was noted with respect to sex (0.78), age (0.28), median duration (0.9), and median dosage (0.9) of ZA. CONCLUSION: Oro-dental screening and dental monitoring shall reduce the incidence of ONJ. Within the limitations of our study, no significant relation could be pointed toward the dosage and duration of the drug and development of ONJ.

Molecular Characterization of the Indigenous Stingless Bees (Tetragonula spp. Complex) Using ISSR Marker from Southern Peninsular India.

India is a country bestowed enormously with stingless bees, but genetic information about them is extremely minimal. This study focused to tap the geographic allocation, genetic variability, and differentiation among Tetragonula species complexes from natural and semi-urban habitats. Genetic analyses were assessed among 36 contrasting genotypes utilizing 20 ISSR primers. The dual combination exquisitely and productively amplified 245 DNA fragments at the loci, of which 240 bands were polymorphic (97.95%). Low to moderate level of genetic differentiation was detected from different estimators (Ht 0.29, G' STest 0.16, D est 0.072, F ST 0.14, and Nm 2.68). Hierarchical clustering analysis aided to partition the individual genotypes into its respective five species group formed, aided by substantial bootstrap support values, but differing under morphological identification. It also provided valuable insight into the moderate eco-genetic diversity (H 0.39) prevailing from geographically scattered inhabitants. Potential exploitation of hyper-variable ISSR marker turned out fairly as a promising technique for finding valid polymorphisms and infers relevant variations. This baseline information enhances our understanding of the genetic status of the indigenous species from the country.

SPINS-IND: Pellet injector for fuelling of magnetically confined fusion systems.

Using a Gifford-McMahon cycle cryocooler based refrigeration system, a single barrel hydrogen pellet injection (SPINS-IND) system is indigenously developed at Institute for Plasma Research, India. The injector is based on a pipe gun concept, where a pellet formed in situ in the gun barrel is accelerated to high speed using high pressure light propellant gas. The pellet size is decided by considering the Greenwald density limit and its speed is decided by considering a neutral gas shielding model based scaling law. The pellet shape is cylindrical of dimension (1.6 mm ℓ × 1.8 mm φ). For pellet ejection and acceleration, a fast opening valve of short opening duration is installed at the breech of the barrel. A three-stage differential pumping system is used to restrict the flow of the propellant gas into the plasma vacuum vessel. Diagnostic systems such as light gate and fast imaging camera (240 000 frames/s) are employed to measure the pellet speed and size, respectively. A trigger circuit and a programmable logic controller based integrated control system developed on LabVIEW enables to control the pellet injector remotely. Using helium as a propellant gas, the pellet speed is varied in the range 650 m/s-800 m/s. The reliability of pellet formation and ejection is found to be more than 95%. This paper describes the details of SPINS-IND and its test results.

Flavored e-cigarette use: Characterizing youth, young adult, and adult users.

The purpose of this study is to investigate how the use of flavored e-cigarettes varies between youth (12-17 years old), young adults (18-29 years old), and older adults (30 + years old). Cross-sectional surveys of school-going youth (n = 3907) and young adult college students (n = 5482) in Texas, and young adults and older adults (n = 6051) nationwide were administered in 2014-2015. Proportions and 95% confidence intervals were used to describe the percentage of e-cigarette use at initiation and in the past 30 days that was flavored, among current e-cigarette users. Chi-square tests were applied to examine differences by combustible tobacco product use and demographic factors. Most e-cigarette users said their first and "usual" e-cigarettes were flavored. At initiation, the majority of Texas school-going youth (98%), Texas young adult college students (95%), and young adults (71.2%) nationwide said their first e-cigarettes were flavored to taste like something other than tobacco, compared to 44.1% of older adults nationwide. Fruit and candy flavors predominated for all groups; and, for youth, flavors were an especially salient reason to use e-cigarettes. Among adults, the use of tobacco flavor at initiation was common among dual users (e-cigarettes + combustible tobacco), while other flavors were more common among former cigarette smokers (P = 0.03). Restricting the range of e-cigarette flavors (e.g., eliminating sweet flavors, like fruit and candy) may benefit youth and young adult prevention efforts. However, it is unclear what impact this change would have on adult smoking cessation.

Same-day light-emitting diode fluorescence microscopy for the diagnosis of tuberculosis in Chhattisgarh, India.

SETTING: Three medical college hospitals using light-emitting diode fluorescence microscopy (LED-FM) for diagnosing tuberculosis (TB) in Chhattisgarh, India. OBJECTIVES: To assess and compare the proportion of sputum smear-positive TB patients diagnosed through same-day microscopy (spot-spot) strategy or with the conventional (spot-morning) strategy. METHODS: During November 2012 - March 2013, all consecutively enrolled presumptive TB patients (aged ≥ 18 years) were requested to provide three specimens: two spot specimens collected 1 h apart on the first day and one early morning specimen the next day; these were stained using auramine-O and examined using LED-FM. RESULTS: Of 1716 (93% of total 1845) presumptive TB patients who provided all three specimens, 218 (13%) were smear-positive: 200 (11.7%) by same-day microscopy and 217 (12.7%) by the conventional method (McNemar's χ(2) 13.5, df 1, P = 0.0002). Eighteen (8.3%) cases were missed by the same-day method. CONCLUSION: Although LED-FM is more sensitive to paucibacillary samples, 8% of smear-positive cases were missed using the same-day method. These findings indicate the need to revisit the global applicability of the current World Health Organization recommendation of switching to same-day diagnosis from the conventional policy.

Studies on growth morphology, UV absorbance and luminescence properties of sulphur doped ZnO nanopowders synthesized by the application of ultrasound with varying input power.

Sulphur doped ZnO nanopowders have been prepared by sonochemical method in continuous (CS) and pulsed (PS) modes. Precipitation time was found to vary with input power of applied ultrasound. X-ray diffraction studies show the formation hexagonal wurtzite phase of nano ZnO but variation in size, strain and lattice parameters was observed in the samples synthesized through CS and PS modes with different input power. Surface morphology of the samples changed significantly with input power of the pulse as observed from scanning electron microscopic results. Well defined and c-axis oriented nanorods and multipodes were observed with power setting of 187.5 W whereas in all other cases no definite morphology was observed. UV absorption studies show higher value of absorbance for ZnO:S. Further, the absorption band becomes sharp for the samples prepared with power setting of 225 W. PL spectra show several bands due to different transitions in the region of 400-780 nm which have been explained in the light of excitonic and defect induced transitions in ZnO.

2-(2,4-Dichloro-phen-yl)-N-(1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-4-yl)acetamide.

In the crystal structure of the title compound, C19H17Cl2N3O2, the mol-ecules form dimers of the R2(2)(10) type through N-H⋯O hydrogen bonding. As a result of steric repulsion, the amide group is rotated with respect to both the dichloro-phenyl and 2,3-dihydro-1H-pyrazol-4-yl rings, making dihedral angles of 80.70 (13) and 64.82 (12)°, respectively. The dihedral angle between the dichloro-phenyl and 2,3-dihydro-1H-pyrazol-4-yl rings is 48.45 (5)° while that between the 2,3-dihydro-1H-pyrazol-4-yl and phenyl rings is 56.33 (6)°.

2-(2,6-Dichloro-phen-yl)-N-(1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-4-yl)acetamide.

In the title compound, C19H17Cl2N3O2, the amide group is planar and, through N-H⋯O hydrogen bonding to an adjoining mol-ecule, forms dimers of the R2(2)(10) type. As a result of steric repulsion, the amide group is rotated with respect to both the dichloro-phenyl and 2,3-dihydro-1H-pyrazol-4-yl rings, making dihedral angles of 71.63 (11) and 57.93 (10)°, respectively. The dihedral angle between the dichloro-phenyl and 2,3-dihydro-1H-pyrazol-4-yl rings is 76.60 (10)° while that between the 2,3-dihydro-1H-pyrazol-4-yl and phenyl rings is 49.29 (7)°. The crystal structure also features weak C-H⋯O inter-actions.