Decoding the genetic influence of CT60 non-coding polymorphism in CTLA-4 gene and sCTLA-4 biomarker with rheumatoid arthritis in the Indian population.

BACKGROUND: Cytoplasmic T Lymphocyte Antigen - 4 (CTLA-4) gene encodes an immunoregulatory receptor expressed on surface of activated T-cells to mediate peripheral tolerance against self-antigen. It suppresses auto-reactive T-cell proliferation either by inactivation or apoptosis of T-cells. The CTLA-4 mRNA undergoes alternative splicing to synthesize a native soluble form of CTLA-4 (sCTLA-4) protein, which lacks exon 3 that encodes for transmembrane region. As a result, sCTLA-4 circulates as a soluble serum protein and acts as an immunoregulator molecule to maintain homeostasis in the blood. MATERIALS AND RESULTS: Techniques coupled with quantitative Polymerase Chain Reaction (qPCR) and High-Resolution Melting Analysis (HRMA) were used to screen CTLA-4 3'Untranslated Region (UTR) CT60 (A/G) rs3087243 Single Nucleotide Polymorphism (SNP) and their association with Rheumatoid Arthritis (RA) in the Indian population. In addition, we also evaluated the concentration of sCTLA-4 serum protein in RA patients carrying rs3087243 SNP with different genotypes (A/A, G/A, and G/G). Statistical analysis of Odds Ratio (OR), Confidence Interval (C.I), and Relative Risk (RR) have shown that frequency of CTLA-4 rs3087243 SNP G/G genotype was significantly associated with RA in the Indian population (OR 1.7140; CI = 1.0765 to 2.7290; RR = 1.5434; p = 0.0232). The sCTLA-4 concentration was also significantly lower in RA patients carrying rs3087243 SNP G/G genotype than control group (p < 0.001). CONCLUSION: Co-inheritance of CTLA-4 signal peptide and 3'UTR SNPs may activate RAPP pathway. Downregulation of CTLA-4 and sCTLA-4 serum protein by rs3087243 SNP can increase the hyperactivation of T-cells, which causes RA.

Knockout of ecdysis triggering hormone receptor (ETHr) gene adversely affects the nymphal molting and adult reproduction in Bemisia tabaci.

Bemisia tabaci (Gennadius) is one of the most dangerous polyphagous pests in the world causing damage to various crops by sucking sap during the nymphal and adult stages. Chemical management of whiteflies is challenging because of the emergence of pesticide resistance. RNA interference has been well established in whitefly to study the functions of various genes. G-protein coupled receptors (GPCRs) are important targets for development of new generation insecticides. In this study, Ecdysis triggering hormone receptor (ETHr) gene expression was recorded in different stages of whitefly and its function has been studied through RNAi. The expression of ETHr is highest in third-instar nymphs followed by other nymphal instars, pupae and newly emerged adults. Silencing of ETHr resulted in significantly higher adult mortality (68.88%), reduced fecundity (4.46 eggs /female), reduced longevity of male and female (1.05 and 1.40 days, respectively) when adults were fed with dsETHr @ 1.0 µg/µl. Silencing of ETHr in nymphs lead to significantly higher mortality (81.35%) as compared to control. This study confirms that ETHr gene is essential for growth and development of whitefly nymphs and adults. Hence, it can be future target for developing dsRNA based insecticides for management of whitefly.

A method for noninvasive beat-by-beat visualization of His bundle signals.

BACKGROUND: Invasive recording of His bundle signals (HBS) in electrophysiological study (EPS) is important in determining HV interval, the time taken to activate the ventricles from the His bundle. Noninvasive surface measurements of HBS are attempted by averaging typically 100-200 cardiac cycles of ECG time series in body surface potential mapping (BSPM) and in magnetocardiography (MCG) which records weak cardiac magnetic fields by highly sensitive detectors. However, noninvasive beat-by-beat extraction of HBS is challenged by ramp-like atrial signals and noise in PR segment of the cardiac cycle. METHODS: By making use of a signal-averaged trace showing prominent HBS as a guide trace, we developed a method combining interval-dependent wavelet thresholding (IDWT) and signal space projection (SSP) technique to eliminate artifacts from single beats. The method was applied on MCG recorded on 21 subjects with known HV intervals based on EPS and noninvasive signal-averaging, including five subjects with BSPM recorded subsequently. The method was also applied on stress-MCG of a subject featuring autonomic dynamics. RESULTS: HBS could be extracted from 19 out of 21 subjects by signal-averaging whose timing differed from EPS between -8 and 11 ms as tested by 2 observers. HBS in single beats were seen as aligned patterns in inter-beat contours and were appreciable in stress-MCG and conspicuous than BSPM. The performance of the method was evaluated on simulated and measured MCG to be adequate if the signal-to-noise ratio was at least 20 dB. CONCLUSIONS: These results suggest the use of this method for noninvasive assessments on HBS.

Bacterial diversity of a floating vegetation (Phumdi) of Loktak Lake and its extracellular enzymes and bacterial antagonistic property.

134 bacterial strains were isolated from phumdis of Loktak Lake. Through 16S rRNA sequencing, Bacillus sp. (23, 17.1%), Staphylococcus sp. (14, 10.4%), Pseudomonas sp. (11, 8.2%) and Acinetobacter sp. (8, 5.9%) were identified as the predominant bacterial taxa of Loktak Lake. B. pumulis (12, 8.9%), S. arlettae (4, 2.9%), P. knackmussii (6, 4.4%) are the leading species of Bacillus, Staphylococcus and Pseudomonas, respectively. Similarly, A. seifertii (2, 1.4%) and A. calcoaceticus (2, 1.4%) are the common species of Acinetobacter. 75 (55.9%) bacterial strains showed the ability to hydrolyze one or more extracellular enzymes tested. Among the extracellular enzymes produced by the bacterial isolates, the presence of elastase activity cannot be underestimated, since the enzyme is involved in the process of bacterial lung infection. Phosphate solubilizing activity could be seen in 11.1% of the bacterial isolates. 27 (20.1%) of the strains shown to have antagonistic activity against one or more tested pathogens. An isolate, MRC 52 showed antagonistic activity against eleven different pathogens including carbapenem resistant E. coli which was further subjected to extraction and identification of the biomolecule exerting the antimicrobial property. Based on GC-MS analysis, the bioactive compound was identified as phenyl ethyl alcohol.

Reliability of Frankfort Horizontal Plane with True Horizontal Plane in Cephalometric Measurements.

INTRODUCTION: The purpose of the study is to determine reliability of true horizontal (TH) plane with Frankfort horizontal (FH) plane in cephalometric measurements. MATERIALS AND METHODS: One hundred lateral cephalograms were collected and traced using FACAD software, all cephalometric measurements which included FH plane as reference planes were used in this study. Cephalometric tracing was done with FH plane as reference plane and another set of measurements with TH plane as reference plane was done. Statistical tests were done using independent sample t-test (p <0.05). RESULTS: Results of an independent t-test revealed a statistically insignificant difference that was observed for all the parameters assessed (p <0.05). Both angular and linear measurements showed an insignificant difference between FH and TH. CONCLUSION: Within the limitations of the study, the results show that the FH plane is closest and as reliable as the TH plane. The variation of FH was least for all the parameters assessed.

Full blood count values in adolescents and its comparison by gender and ethnicity in Seremban district, Malaysia.

BACKGROUND: Adolescence is when an individual undergoes development and growth. Many studies suggest variations in the number and size of blood cells during this period in various individuals. The full blood count (FBC) is often the starting point of medical investigations, which help diagnose a wide range of illnesses, infections, and diseases. This study aimed to report the mean FBC values and compare them by gender and ethnicity, using blood results from the thalassemia screening programme in Seremban District, Malaysia. MATERIALS AND METHODS: This cross-sectional study used secondary data from the thalassemia screening programme on Form 4 students aged 15-16 years from January 2018 to October 2018 by the Seremban District Health Office, Malaysia. These students participated in the thalassemia screening programme in which their blood samples were taken for FBC analysis. The data were extracted for this study. RESULTS: There were statistically significant gender-based differences for total white blood cell (WBC) count, neutrophils, lymphocytes, mixed WBC, and platelets. It was also observed that ethnic-specific differences were statistically significant for RBC count, platelets, platelet distribution width and mean platelet volume. CONCLUSION: This study was able to report the mean FBC values among Malaysian adolescents with respect to their gender and ethnicity, of which there is a lack of published data.

An intuitionistic fuzzy decision support system for COVID-19 lockdown relaxation protocols in India.

In January 2020, the World Health Organization (WHO) identified a world-threatening virus, SARS-CoV-2. To diminish the virus spread rate, India implemented a six-month-long lockdown. During this period, the Indian government lifted certain restrictions. Therefore, this study investigates the efficacy of India's lockdown relaxation protocols using fuzzy decision-making. The decision-making trial and evaluation laboratory (DEMATEL) is one of the fuzzy MCDM methods. When it is associated with intuitionistic fuzzy circumstances, it is known as the intuitionistic fuzzy DEMATEL (IF-DEMATEL) method. Moreover, converting intuitionistic fuzzy into a crisp score (CIFCS) algorithm is an aggregation technique utilized for the intuitionistic fuzzy set. By using IF-DEMATEL and CIFCS, the most efficient lockdown relaxation protocols for COVID-19 are determined. It also provides the cause and effect relationship of the lockdown relaxation protocols. Additionally, the comparative study is carried out through various DEMATEL methods to see the effectiveness of the result. The findings would be helpful to the government's decision-making process in the fight against the pandemic.

Efficacy of short-synthetic antifungal peptides on pathogenic Aspergillus flavus.

The efficacies of three short synthetic antifungal peptides were tested for their inhibitory action on pathogenic fungi, Aspergillus flavus. The sequences of the short synthetic peptides are PPD1- FRLHF, 66-10-FRLKFH, 77-3- FRLKFHF, respectively. These test peptides inhibited fungal growth and showed a membranolytic activity. The fungal biomass and ergosterol levels were significantly low in peptides treated samples. Further, the fungal cell wall component chitin was also found to be lower in peptides treated samples. Scanning electron microscopic images also showed highly wrinkled fungal mycelia. Significant membrane permeabilisation as well as potassium ion leakage was also observed in fungal samples treated with peptides. To assess the membrane damage, the uptake of Sytox green dye was employed. At tested concentration, peptides induced fungal membrane damage as evidenced by the green fluorescence. Further, at tested concentration, these peptides induced an oxidative stress in A.flavus as evidenced by an increase in the ROS production, malondialdehyde levels, increase in the antioxidant enzymes - superoxide dismutase, catalase with concomitant decrease in the reduced glutathione content. Additionally, a growth dependent reduction in aflatoxin levels were also observed in peptides treated samples. Docking studies on the interaction of the peptides with a trans-membrane protein calcium ATPase of A. flavus showed that all the peptides were able to bind to the protein with high z rank score. The activity of the calcium ATPase was significantly decreased in peptides treated fungal samples, thereby validating the docking results. Among all the tested peptides, 77-3 peptide exhibited the maximal membrane damage property.

Polyphenols as an Effective Therapeutic Intervention Against Cognitive Decline During Normal and Pathological Brain Aging.

Research in animals and humans has indicated that polyphenols can delay the age-related decline in learning, memory and neurodegenerative diseases. Among the polyphenols, berry phenolics have extensive beneficial effects because of their antioxidant and anti-inflammatory properties. Long-term consumption of grapes results in accumulation of polyphenols in the brain, which modulates cell-signalling pathways and neutralises the redox imbalance in the aging brain. Here we review the in vivo and in vitro evidence for considering grape-derived polyphenolics, the flavonoids- catechins, epicatechin, anthocyanidin, and quercetin, and non-flavonoids-gallic acid and resveratrol, as effective dietary sources to facilitate cognition in adults and lessen the decline in the old and pathogenic states, Alzheimer's and Parkinson's disease. Furthermore, a combined intervention of polyphenols along with regular physical exercise provides cognitive benefits for the aging brain and holds promising venues for preclinical and clinical studies in formulating neuro-nutraceuticals as functional foods for a healthy brain.

Nitrogen-doped carbon quantum dots conjugated isoreticular metal-organic framework-3 particles based luminescent probe for selective sensing of trinitrotoluene explosive.

Amine group-containing isoreticular metal-organic framework (IRMOF-3) particles are utilized for the first time as a trinitrotoluene (TNT) sensing material. IRMOF-3 particles are synthesized using zinc nitrate as a metal precursor and 2-amino-1,4-benzenedicarboxylic acid as a linker. The nitrogen-doped carbon quantum dots (NCQDs) are synthesized from citric acid and ethylenediamine as carbon and nitrogen precursor, respectively. The NCQDs are conjugated with IRMOF-3 particles as IRMOF-3/NCQDs. The TEM micrograph revealed the average size of IRMOF-3 particles to be 363.66 nm. The photoluminescence emission intensity of IRMOF-3 particles at λem 430 nm is highly increased in the presence of NCQDs (λex 330 nm). Both the as-synthesized IRMOF-3 and IRMOF-3/NCQD particles are explored for TNT detection to compare the effect of NCQDs on the IRMOF-3 particle surface. Lower limit of detection (7.5 × 10-8 M) and higher Stern-Volmer constant (4.46 × 106 M-1) are achieved by IRMOF-3/NCQD particles. The association constant also increased from 5.3 × 104 to 2.78 × 106 M-1 after the conjugation of IRMOF-3 particles with NCQDs. Moreover, enhanced selectivity for TNT over trinitrophenol is achieved using the IRMOF-3/NCQD particles. Graphical Abstract.

Antibiotic resistance and virulence genes in nisin-resistant Enterococcus faecalis isolated from raw buffalo milk modulate the innate functions of rat macrophages.

AIMS: To elucidate the antibiotic resistance and virulence genes of nisin-resistant Enterococcus faecalis isolated from raw buffalo milk and to study the effect of nisin-sensitive and -resistant E. faecalis on the innate immunity of rats. METHODS AND RESULTS: Slanetz-Bartley agar plates containing nisin were used to isolate nisin-resistant E. faecalis. The virulence factors were ascertained using quantitative real-time polymerase chain reaction. Cell viability, phagocytosis, intracellular survival and enzyme assays were performed to investigate the interaction of E. faecalis with rat macrophages. Nisin-resistant E. faecalis was less prone to phagocytosis and survived longer inside the macrophages, due to reduced production of reactive oxygen species and nitric oxide. The viability and activation of macrophages was also reduced in the presence of resistant E. faecalis, as observed by enhanced lactate dehydrogenase production and reduced ß-galactosidase. CONCLUSIONS: Nisin-resistant E. faecalis and its virulence factors were reported in raw buffalo milk. This study shows that nisin-resistant variants exhibited cross resistance to antibiotics and suppressed the innate immune responses of rats by directly affecting macrophage activity. SIGNIFICANCE AND IMPACT OF THE STUDY: This study elucidated the contamination of raw buffalo milk by nisin-resistant E. faecalis, which may pose food safety risk.

Susceptibility profile, resistance mechanisms & efficacy ratios of fosfomycin, nitrofurantoin & colistin for carbapenem-resistant Enterobacteriaceae causing urinary tract infections.

Background & objectives: The escalation in carbapenem resistance among Enterobacteriaceae has resulted in a lack of effective therapeutic alternatives. Older antimicrobials, fosfomycin, nitrofurantoin and colistin for urinary tract infections (UTIs) caused by carbapenem-resistant Enterobacteriaceae (CRE) may be effective treatment options. The objectives of this study were to evaluate the utility of fosfomycin, nitrofurantoin and colistin in treating UTI caused by CRE and molecular characterization of the plasmid-mediated carbapenem resistance mechanisms. Methods: Consecutive, non-duplicate isolates of CR Escherichia coli and Klebsiella spp. from urine cultures were included (n=150). Minimum inhibitory concentrations (MIC) were determined by E-test (fosfomycin and nitrofurantoin) and broth microdilution (colistin). Efficacy ratios were derived by dividing susceptibility breakpoints by observed MIC values of the drugs for the isolates. Isolates were screened for genes coding for carbapenemases using multiplex PCR. Fosfomycin, nitrofurantoin and colistin-resistant isolates were screened for plasmid-borne resistance genes fos A3, oqx AB and mcr-1, respectively using PCR. Results: Among E. coli, 98.9, 56 and 95 per cent isolates were susceptible to fosfomycin, nitrofurantoin and colistin, respectively, while 94 and 85 per cent of Klebsiella spp. were susceptible to fosfomycin and colistin, respectively. The efficacy ratios indicated fosfomycin as the drug of choice for UTI caused by CR E. coli and Klebsiella spp., followed by colistin. The blaNDM gene was most common, followed by blaOXA48-like. Plasmid-borne genes encoding resistance to fosfomycin, nitrofurantoin and colistin were absent. Interpretation & conclusions: With increasing resistance against the current treatment options, older drugs may emerge as effective options. Molecular screening of resistant isolates is essential to prevent the spread of plasmid-borne resistance against these drugs.

Development of Cryopreservation Protocol for Aquilaria malaccensis Lam., a Recalcitrant Seeded Tropical Tree Species.

BACKGROUND: Cryopreservation is the only possible process for long-term germplasm storage of species with recalcitrant behavior. Aquilaria malaccensis is a recalcitrant seeded tropical tree that produces a distinctive fragrance which has high value in the commercial market. OBJECTIVES: In the present study, we aimed to develop possible long-term storage techniques for A. malaccensis germplasm. MATERIALS AND METHODS: We used zygotic embryos and in vitro derived nodal buds as explants. The experiments were performed based on dehydration and encapsulation-dehydration methods. RESULTS: When the dehydration technique was applied, survival (43%) and regeneration (23%) was found to be higher for zygotic embryos than for in vitro derived nodal buds (13% & 10% respectively). In addition to moisture content within the tissue during cryogen exposure, dehydration duration has an important contributory role in post cryo-survival and regeneration. A slight increase in survival (47%) and regeneration (30%) were observed in zygotic embryos with a modification to rehydration after re-warming, while such a change did not improve success with in vitro derived nodal buds. In contrast, the encapsulation-dehydration technique was found to be more effective for in vitro derived nodal buds than zygotic embryos. All the encapsulated nodal buds that survived (27%) regenerated into plantlets while encapsulated zygotic embryos failed to regenerate into plantlets. CONCLUSION: This is the first report on the cryopreservation of A. malaccensis and the developed protocol conveys a comprehensive idea of its reliability for the long-term storage of this desiccation sensitive (recalcitrant) seeded tree species.

The Kuala Pilah cluster cataract study: Accessible eye care reduces cataract blindness.

OBJECTIVE: The Ministry of Health introduced the cluster hospital project in Kuala Pilah district in 2016 to allow sharing of resources between the hospitals in the same vicinity. The aim of this study is to compare the demographic profile, prevalence of cataract blindness and low vision among patients who presented for cataract surgery before and after the programme. METHODOLOGY: This is a retrospective cohort study of patients who underwent cataract surgery in Kuala Pilah Cluster Hospitals between 2010 and 2017. A total of 2539 records of patients were reviewed. Patients were assigned into two groups: Group 1 (2010-2012)- before the programme (2010-2012) and Group 2 (2015-2017) after the introduction of the programme. RESULTS: There was a significant increase in number of cataract cases in the district hospital after the cluster initiative. The mean age of patients undergoing cataract surgery was similar in both groups. The common comorbidities were hypertension (Group 1=57.3%; Group 2=70.8%) and diabetes mellitus (Group 1=40.6%; Group 2=51.1%). In 2010-2012, most of the patients were one eye blind (34.4%), whereas in 2015-2017 majority of patients presented with vision better than 6/18 (43.5%). The proportion of patients with cataract blindness reduced from 6% in 2010-2012 to 4.3% in 2015-2017 (p<0.01). CONCLUSION: There is a significant decrease in percentage of patients with cataract blindness and low vision after the introduction of Kuala Pilah Cluster Hospital Program. We believe that that cluster hospital system is effective in improving accessibility to eye care and therefore increases the cataract detection rate.

Phenazine-1-carboxylic acid and soil moisture influence biofilm development and turnover of rhizobacterial biomass on wheat root surfaces.

Phenazine-1-carboxylic acid (PCA) is produced by rhizobacteria in dryland but not in irrigated wheat fields of the Pacific Northwest, USA. PCA promotes biofilm development in bacterial cultures and bacterial colonization of wheat rhizospheres. However, its impact upon biofilm development has not been demonstrated in the rhizosphere, where biofilms influence terrestrial carbon and nitrogen cycles with ramifications for crop and soil health. Furthermore, the relationships between soil moisture and the rates of PCA biosynthesis and degradation have not been established. In this study, expression of PCA biosynthesis genes was upregulated relative to background transcription, and persistence of PCA was slightly decreased in dryland relative to irrigated wheat rhizospheres. Biofilms in dryland rhizospheres inoculated with the PCA-producing (PCA+ ) strain Pseudomonas synxantha 2-79RN10 were more robust than those in rhizospheres inoculated with an isogenic PCA-deficient (PCA- ) mutant strain. This trend was reversed in irrigated rhizospheres. In dryland PCA+ rhizospheres, the turnover of 15 N-labelled rhizobacterial biomass was slower than in the PCA- and irrigated PCA+ treatments, and incorporation of bacterial 15 N into root cell walls was observed in multiple treatments. These results indicate that PCA promotes biofilm development in dryland rhizospheres, and likely influences crop nutrition and soil health in dryland wheat fields.

Identification and functional characterization of antimicrobial peptide from the marine crab Dromia dehaani.

Many naturally occurring biologically active compounds are derived from marine organisms. Brachyuran crab Dromia dehaani is also known as sponge crab. Hence, this study was undertaken to isolate and characterize the antimicrobial peptide from the marine crab hemolymph. Therefore the objectives of this research work was to high light the functions of biologically active peptide Dromidin. Dromidin was identified and confirmed as a new antimicrobial peptide in the hemolymph of the crab D. dehaani. Antimicrobial peptide was characterized and confirmed by using various analytical techniques. ESI-MS analysis of the active antimicrobial hemolymph fraction resulted in the value 513.0 Da. In addition, current findings strongly suggest that this peptide is functionally important against the pathogens.

Simultaneous quantitative monitoring of four indicator contaminants of emerging concern (CEC) in different water sources of Central India using SPE/LC-(ESI)MS-MS.

Environmental occurrence of CECs poses a great threat to both aquatic life and human health. The aim of this study was to optimize and validate SPE/LC-(ESI)MS-MS method for simultaneous quantitative monitoring of two sub-classes of CECs (pharmaceuticals and hormones) and to estimate the concentrations of select CECs in environmental water samples. For all the tested analytes, recoveries in laboratory reagent water were greater than 81%. Average percent (relative standard deviation) RSD of the analytes in recovery, repeatability, and reproducibility experiments were ≤ 10%. Determination coefficients (r2) of primidone, diclofenac, testosterone, and progesterone were estimated to be 0.9979, 0.9972, 0.9968, and 0.9962, respectively. Limits of detection (LOD) for primidone, diclofenac, testosterone, and progesterone were 4.63 ng/L, 5.36 ng/L, 0.55 ng/L, and 0.88 ng/L, respectively. Limits of quantification (LOQ) for primidone, diclofenac, testosterone, and progesterone were 14.72 ng/L, 17.06 ng/L, 1.766 ng/L, and 2.813 ng/L, respectively. Average recoveries in environmental water and wastewater samples were greater than 74% and RSD were ≤ 7%. Trace levels (68.33-125.70 ng/L) of primidone were detected in four environmental water samples, whereas diclofenac was not detected in any of the tested sample. Trace levels of progesterone were observed in two environmental samples (16.64 -203.73 ng/L), whereas testosterone was detected in STP inlet sample (178.16 ng/L).

Targeted Biopsy to Detect Gleason Score Upgrading during Active Surveillance for Men with Low versus Intermediate Risk Prostate Cancer.

PURPOSE: We sought to determine the rate of upgrading to Gleason score 4 + 3 or greater using targeted biopsy for diagnosis and monitoring in men undergoing active surveillance of prostate cancer. MATERIALS AND METHODS: Study subjects comprised all 259 men, including 196 with Gleason score 3 + 3 and 63 with Gleason score 3 + 4, who were diagnosed by magnetic resonance imaging/ultrasound fusion guided biopsy from 2009 to 2015 and underwent subsequent fusion biopsy for as long as 4 years of active surveillance. The primary end point was the discovery of Gleason score 4 + 3 or greater prostate cancer. Followup biopsies included targeting of positive sites, which were tracked in an Artemis™ device. Kaplan-Meier curves were generated to determine upgrading rates, stratified by initial Gleason score and prostate specific antigen density. RESULTS: Based on a Cox proportional hazard model, men with Gleason score 3 + 4 were 4.65 times more likely to have upgrading than men with an initial Gleason score of 3 + 3 at 3 years (p <0.01). By the third surveillance year 63% of men with Gleason score 3 + 4 had been upgraded compared with 18.0% who started with Gleason score 3 + 3 (p <0.01). Of all 33 upgrades 32 (97%) occurred at a magnetic resonance imaging visible or a tracked site of tumor, rather than at a previously negative systematic site. Independent predictors of upgrading were Gleason score 3 + 4, prostate specific antigen density 0.15 ng/ml/cm3 or greater and a grade 5 lesion on magnetic resonance imaging. The incidence rate ratio of upgrading (Gleason score 3 + 4 vs 3 + 3) was 4.25 per year of patient followup (p <0.01). CONCLUSIONS: During active surveillance of prostate cancer, targeting of tracked tumor foci by magnetic resonance imaging/ultrasound fusion biopsy allows for heightened detection of Gleason score 4 + 3 or greater cancers. Baseline variables directly related to important upgrading that warrant increased vigilance include Gleason score 3 + 4, prostate specific antigen density 0.15 ng/ml/cm3 or greater and grade 5 lesions on magnetic resonance imaging.

Application of cell-based assays for toxicity characterization of complex wastewater matrices: Possible applications in wastewater recycle and reuse.

Exposure to pre-concentrated inlet or outlet STP wastewater extracts at different concentrations (0.001% to 1%) induced dose-dependent toxicity in MCF-7 cells, whereas drinking water extracts did not induce cytotoxicity in cells treated. GC-MS analysis revealed the occurrence of xenobiotic compounds (Benzene, Phthalate, etc.) in inlet/outlet wastewater extracts. Cells exposed to inlet/outlet extract showed elevated levels of reactive oxygen species (ROS: inlet: 186.58%, p<0.05, outlet, 147.8%, p<0.01) and loss of mitochondrial membrane potential (Δψm: inlet, 74.91%, p<0.01; outlet, 86.70%, p<0.05) compared to the control. These concentrations induced DNA damage (Tail length: inlet: 34.4%, p<0.05, outlet, 26.7%, p<0.05) in treated cells compared to the control (Tail length: 7.5%). Cell cycle analysis displayed drastic reduction in the G1 phase in treated cells (inlet, G1:45.0%; outlet, G1:58.3%) compared to the control (G1:67.3%). Treated cells showed 45.18% and 28.0% apoptosis compared to the control (1.2%). Drinking water extracts did not show any significant alterations with respect to ROS, Δψm, DNA damage, cell cycle and apoptosis compared to the control. Genes involved in cell cycle and apoptosis were found to be differentially expressed in cells exposed to inlet/outlet extracts. Herein, we propose cell-based toxicity assays to evaluate the efficacies of wastewater treatment and recycling processes.

SEM Analysis of Residual Dentin Surface in Primary Teeth Using Different Chemomechanical Caries Removal Agents.

BACKGROUND: The purpose of this in vitro study was to analyze the residual dentinal surfaces following caries removal using two chemomechanical methods (Papacarie Duo and Carie Care), by scanning electron microscopy (SEM). STUDY DESIGN: Twenty extracted primary molars with active occlusal carious lesions were randomly assigned two groups depending on the CMCR agent used for the caries excavation - Group 1 - with Papacarie Duo and Group - 2 with Carie Care. After the caries excavation, the specimens were subjected to SEM analysis. RESULTS: Though both the agents showed the minimal smear layer with the patent dentinal tubules, Carie care showed patent dentinal tubules with a clearly exposed peritubular and intertubular collagen network. CONCLUSION: Carie Care treated surface exhibited better surface morphology of residual dentin.