A hospital-based study of prostate biopsy results in Indian males.

Introduction: The prostate is a gland belonging to the male reproductive system. Aging results in the dysfunction of the prostate that may present as inflammation, enlargement, and cancer. Additionally, the diseases of the prostate including cancers are slow in progression, and therefore, it is difficult to diagnose them early. Hence, it is increasingly important for physicians to recommend histopathological examination of the prostate gland to identify, manage, and treat prostate cancers. This study was conducted to assess prostate diseases among biopsy specimen collected from patients with signs of prostate diseases. Materials and Methods: This prospective study was conducted in the Department of Pathology, Deccan College of Medical Sciences, Owaisi Hospital, Hyderabad, between June 2012 and September 2014. All gross specimens (n = 300) of the prostate such as the needle biopsies of the prostate, transurethral resection of the prostate (TURP) chips, and excised specimens of the prostate were included in the study. Histopathological examinations of the biopsies were performed for nuclear size, chromatin material, nucleoli, membrane thickness, irregularity, cytoplasmic granularity, staining, and cell border conspicuity. The biopsies were also assessed for lobule formation, secretions, polymorphonuclear leukocytes, lymphocytes, macrophages, connective tissue stromal cells, their arrangements, and acellular connective tissue material. Results: Of 300 total prostatic biopsies performed, 56 (18.66%) were identified as inflammatory lesions of the prostate (prostatitis), 98 (32.66%) revealed benign prostatic lesions (benign prostatic hyperplasia (BPH)), 112 (37.33%) were identified as BPH with premalignant lesions, and 34 (11.33%) were revealed as malignant tumors of the prostate. Chronic prostatitis (67.85%) was the common inflammatory lesion. The majority (91.42%) revealed epithelial lesions compared to stromal lesions (08.58%). BPH was predominantly (28.00%) noticed among patients in the age group of 61-70 years. Prostatic intraepithelial neoplasia (PIN) was observed majorly (53.35%) in the age group of 61-70 years. Most of the prostatic cancers were identified as adenocarcinomas. However, three variants were also categorized as small cell carcinoma, signet ring cell carcinoma, and transitional cell carcinomas. Conclusions: The results reveal that prostatic adenocarcinomas are predominant among the study population. Additionally, prostatic diseases including cancer are commonly noticed among people belonging to the age group of 61-70 years. More than one-third of patients showed BPH with premalignant lesions, and a majority of the study population showed evidence of chronic prostatitis.

An improved class of estimators for estimation of population distribution functions under stratified random sampling.

The main objective of the current study is to suggest an enhanced family of log ratio-exponential type estimators for population distribution function (DF) using auxiliary information under stratified random sampling. Putting different choices in our suggested generalized class of estimators, we found some Specific estimators. The bias and MSE expressions of the estimators have been approximated up to the first order. By using the actual and simulated data sets, we measured the performance of estimators. Based on the results, the suggested estimators for DF show better performance as compared to the preliminary estimators considered here. The suggested estimators have a advanced efficiency than the other estimators examined with the estimators F‾ˆlogPR(st)2, and F‾ˆlogPR(st)4 for both the actual and simulated data sets. The magnitude of the improvement in efficiency is noteworthy, indicating the superiority of the proposed estimators in terms of MSE.

Mutation analysis of BCR-ABL1 kinase domain in chronic myeloid leukemia patients with tyrosine kinase inhibitors resistance: a Malaysian cohort study.

OBJECTIVE: Mutational analysis of BCR::ABL1 kinase domain (KD) is a crucial component of clinical decision algorithms for chronic myeloid leukemia (CML) patients with failure or warning responses to tyrosine kinase inhibitor (TKI) therapy. This study aimed to detect BCR::ABL1 KD mutations in CML patients with treatment resistance and assess the concordance between NGS (next generation sequencing) and Sanger sequencing (SS) in detecting these mutations. RESULTS: In total, 12 different BCR::ABL1 KD mutations were identified by SS in 22.6% (19/84) of patients who were resistant to TKI treatment. Interestingly, NGS analysis of the same patient group revealed an additional four different BCR::ABL1 KD mutations in 27.4% (23/84) of patients. These mutations are M244V, A344V, E355A, and E459K with variant read frequency below 15%. No mutation was detected in 18 patients with optimal response to TKI therapy. Resistance to TKIs is associated with the acquisition of additional mutations in BCR::ABL1 KD after treatment with TKIs. Additionally, the use of NGS is advised for accurately determining the mutation status of BCR::ABL1 KD, particularly in cases where the allele frequency is low, and for identifying mutations across multiple exons simultaneously. Therefore, the utilization of NGS as a diagnostic platform for this test is very promising to guide therapeutic decision-making.

A Serious Game ("Fight With Virus") for Preventing COVID-19 Health Rumors: Development and Experimental Study.

BACKGROUND: Health rumors arbitrarily spread in mainstream social media on the internet. Health rumors emerged in China during the outbreak of COVID-19 in early 2020. Many midelders/elders (age over 40 years) who lived in Wuhan believed these rumors. OBJECTIVE: This study focused on designing a serious game as an experimental program to prevent and control health rumors. The focus of the study was explicitly on the context of the social networking service for midelders/elders. METHODS: This research involved 2 major parts: adopting the Transmission Control Protocol model for games and then, based on the model, designing a game named "Fight With Virus" as an experimental platform and developing a cognitive questionnaire with a 5-point Likert scale. The relevant variables for this experimental study were defined, and 10 hypotheses were proposed and tested with an empirical study. In total, 200 participants were selected for the experiments. By collecting relevant data in the experiments, we conducted statistical observations and comparative analysis to test whether the experimental hypotheses could be proved. RESULTS: We noted that compared to traditional media, serious games are more capable of inspiring interest in research participants toward their understanding of the knowledge and learning of health commonsense. In judging and recognizing the COVID-19 health rumor, the test group that used game education had a stronger ability regarding identification of the rumor and a higher accuracy rate of identification. Results showed that the more educated midelders/elders are, the more effective they are at using serious games. CONCLUSIONS: Compared to traditional media, serious games can effectively improve midelders'/elders' cognitive abilities while they face a health rumor. The gameplay effect is related to the individual's age and educational background, while income and gender have no impact.

Development and Validation of the Observation Checklist Assessing the Hygiene and Sanitation of the Food Preparation Areas in Kota Bharu Kelantan Preschool.

INTRODUCTION: The current inspection checklist for the assessment of food preparation areas in preschools in Malaysia has not been revised since 2012. The checklist's content needs to be improved to ensure relevant parameters are covered during preschool inspections. OBJECTIVE: The objective of this study was to develop and validate an observation checklist for assessing the hygiene and sanitation of food preparation areas in preschools. METHODOLOGY: The study was conducted in Kota Bharu Kelantan from March 2021 to February 2022. The development of the observation checklist was conducted in four stages: (1) the construction of domains and items from the existing literature, (2) content validation by six experts (using the item-level content validity index (I-CVI) and the scale-level content validity index (S-CVI), (3) face validation by 10 experts (using the item-level face validity index (I-FVI) and the scale-level face validity index (S-FVI)), and (4) reliability analysis (using the intercorrelation coefficient (ICC)). Four assessors performed the reliability analysis at two preschools. RESULTS: The initial draft of the checklist contained three domains and 57 items: building and facility (10 subdomains and 38 items), process control (four subdomains and 12 items), and food handlers (one subdomain and seven items). The I-CVI scores for building and facility, process control, and food handlers were 0.97, 1.00, and 1.00, respectively, indicating good relevancy of items. The S-CVI value was 1.0 for all domains, showing good relevance of the items. The I-FVI above 0.8 and S-FVI values above 0.9 for all domains imply that the participants easily understood the checklist. The ICC for each domain was 0.847 (95% CI 0.716-0.902) for the building facility and 1.0 for process control and food handler, and the ICC for the three domains combined was 0.848 (95% CI 0.772-0.904). The final validated checklist consists of three domains with 57 items. CONCLUSION: The newly developed observation checklist is a valid and reliable tool for assessing the hygiene and sanitation of preschool food preparation areas.

Resurgence of Dengue Virus Serotype 4 in Malaysia: A Comprehensive Clinicodemographic and Genomic Analysis.

Dengue virus serotype 4 (DENV-4) has been the rarest circulating serotype in Malaysia, resulting in it being an understudied area. A recent observation from institutional surveillance data indicated a rapid increase in DENV-4-infected cases. The present study aimed to investigate the resurgence of DENV-4 in relation to the demographic, clinical and genomic profiles of 75 retrospective dengue samples. First, the demographic and clinical profiles obtained between 2017 and July 2022 were statistically assessed. Samples with good quality were subjected to full genome sequencing on the Illumina Next Seq 500 platform and the genome data were analysed for the presence of mutations. The effect of the mutations of interest was studied via an in silico computational approach using SWISS-MODEL and AlphaFold2 programs. The predominance of DENV-4 was discovered from 2021 to 2022, with a prevalence of 64.3% (n = 9/14) and 89.2% (n = 33/37), respectively. Two clades with a genetic divergence of 2.8% were observed within the dominant genotype IIa. The majority of DENV-4-infected patients presented with gastrointestinal symptoms, such as vomiting (46.7%), persistent diarrhoea (30.7%) and abdominal pain (13.3%). Two mutations, His50Tyr and Pro144Ser, located at the wing domain of the NS1 protein were discovered to be unique to the recently sequenced DENV-4.

Genotype, phenotype and treatment outcomes of 17 Malaysian patients with infantile-onset Pompe disease and the identification of 3 novel GAA variants.

BACKGROUND: Pompe disease is a rare glycogen storage disorder caused by deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA), leading to glycogen deposition in multiple tissues. Infantile-onset Pompe disease (IOPD) patients present within the first year of life with profound hypotonia and hypertrophic cardiomyopathy. Treatment with enzyme replacement therapy (ERT) has significantly improved survival for this otherwise lethal disorder. This study aims to describe the clinical and molecular spectrum of Malaysian IOPD patients, and to analyze their long term treatment outcomes. METHODS: Seventeen patients diagnosed with IOPD between 2000 and 2020 were included in this retrospective cohort study. Clinical and biochemical data were collated and analyzed using descriptive statistics. GAA enzyme levels were performed on dried blood spots. Molecular analysis of the GAA gene was performed by polymerase chain reaction and Sanger sequencing. Structural modelling was used to predict the effect of the novel mutations on enzyme structure. RESULTS: Our cohort had a median age of presentation of 3 months and median age of diagnosis of 6 months. Presenting features were hypertrophic cardiomyopathy (100%), respiratory insufficiency (94%), hypotonia (88%), failure to thrive (82%), feeding difficulties (76%), and hepatomegaly (76%). Fourteen different mutations in the GAA gene were identified, with three novel mutations, c.1552-14_1552-1del, exons 2-3 deletion and exons 6-10 deletion. The most common mutation identified was c.1935C > A p.(D645E), with an allele frequency of 33%. Sixteen patients received ERT at the median age of 7 months. Overall survival was 29%. Mean age of death was 17.5 months. Our longest surviving patient has atypical IOPD and is currently 20 years old. CONCLUSIONS: This is the first study to analyze the genotype and phenotype of Malaysian IOPD patients, and has identified the c.1935C > A p.(D645E) as the most common mutation. The three novel mutations reported in this study expands the mutation spectrum for IOPD. Our low survival rate underscores the importance of early diagnosis and treatment in achieving better treatment outcomes.

Evaluation of the efficacy of topical Terminalia chebula Retz. with vinegar in the treatment of tinea corporis: a non-inferiority randomized controlled trial.

OBJECTIVES: Unani physicians have suggested a wide range of anti-dermatophytic remedies, although the scientific evidence is scarce. Thus, the efficacy and safety of Terminalia chebula Retz. fruit powder mixed with vinegar was compared with terbinafine hydrochloride 1% cream in the treatment of tinea corporis in order to establish the non-inferiority of test drugs. METHODS: The primary outcome measures were change in the presence or absence of hyphae on KOH mount test, change in pruritus severity assessed on 100 mm VAS and change in physician's global assessment. Secondary outcome measure was change in the dermatology life quality index (DLQI). Hemograms, serum creatinine, serum bilirubin, and random blood sugar levels were measured at the baseline and after treatment to ensure the safety of the interventions. RESULTS: A per-protocol analysis was done on 40 participants (21 in the test group and 19 in the control group). The observed differences in the primary and secondary outcomes between the test and control groups were greater than the non-inferiority margin, signifying that the test drugs were not inferior. CONCLUSIONS: It may be inferred that the trial drug Terminalia chebula Retz. fruit powder mixed with vinegar is not inferior to terbinafine hydrochloride cream in the treatment of tinea corporis.

A bifunctionalised Pb-based MOF for iodine capture and dye removal.

A 2-dimensional Pb(II) metal-organic framework, [Pb(bdc)0.5(py-Phen)NO3]n (SM-3), was synthesized under solvothermal conditions using a mixed ligand approach. SM-3 was assembled using dinuclear SBUs [Pb2(COO)2]2-, an oxygen donor H2bdc = 1,4-benzene dicarboxylic acid, and nitrogen donor py-Phen = pyrazino[2,3-f][1,10]-phenanthroline linkers. SM-3 was characterized by elemental analysis, FT-IR, powder-X-ray diffraction, thermal gravimetric analysis, SEM, EDS, TEM, and single-crystal X-ray diffraction techniques. Crystallographic studies confirmed that SM-3 displays a 2D layered structure with unique anagostic (Pb⋯H) interactions. Interestingly, the presence of abundant π-electron-rich rings embellished with free -N donor sites in the framework makes SM-3 an excellent adsorbent that exhibits adsorption performance for iodine and dyes. The experimental results show that SM-3 reversibly adsorbs radioactive iodine in the solution and vapor phases and exhibits selective adsorption performance for hazardous cationic dyes, namely, methylene blue (MB) and rhodamine-B (Rh-B), from aqueous solution. Moreover, the possible mechanism of iodine and dye adsorption was also discussed in detail. Thus, this work is remarkable for coordination chemists to engineer layered MOFs for adsorption purposes and expands their potential characteristics by converting them into 2D MOF nanosheets to further enhance the adsorption of hazardous pollutants for environment protection.

Application of Multilocus Sequence Typing for the Characterization of Leptospira Strains in Malaysia.

Leptospirosis is a common zoonotic disease in tropical and subtropical countries. It is considered an emerging disease in Malaysia and is a notifiable disease. This study was conducted to characterize Malaysian isolates from human, animal and environmental samples via MLST and rrs2 sequencing in an attempt to develop a Malaysian genotypic database. An existing polymerase chain reaction (PCR)-based MLST scheme was performed to facilitate subsequent sequencing. Out of 46 extracted DNA, 36 had complete MLST profiles whereby all six genes were amplified and sequenced. Most of the pathogenic Leptospira genotypes with full MLST profiles were L. interrogans serogroup Bataviae (n = 17), followed by L. borgpetersenii serogroup Javanica (n = 9), L. interrogans serogroup Sejroe (n = 2), L. interrogans serogroup Australis (n = 2), L. kirschneri (n = 2), L. interrogans serogroup Grippotyphosa (n = 1) and L. interrogans serogroup Pyrogenes (n = 3). Two samples (R3_SER/17 and R4_SER/17) were not closely related with any of the reference strains. For the samples with incomplete MLST profiles, leptospiral speciation was conducted through rrs2 analysis, in which four samples were identified as L. borgpetersenii, five samples were closely related to L. kmetyi and one sample was known as L. yasudae. This study shows that molecular approaches that combine both MLST and rrs2 sequencing have great potential in the comprehensive characterization of pathogenic Leptospira because they can be performed directly from cultured and clinical samples.

A non-inferiority randomized controlled clinical trial comparing Unani formulations and PUVAsol in non-segmental vitiligo.

OBJECTIVES: Greco-Arab medicine is an ancient system of medicine with greater treasure on therapeutics of vitiligo. The trial Unani formulations have not been scientifically explored for their safety and efficacy, but have been repeatedly prescribed by the great Unani physicians in the management of Baras (vitiligo). Hence, these interventions were selected for the trial. METHODS: In this randomized, controlled, open-label clinical trial, 82 participants with non-segmental vitiligo aged 18-40 years were block randomized to either receive Unani interventions or control for 16 weeks. Out of 82 participants, 42 were randomized to the Unani group and 40 were randomized to the control group. The primary outcome measure was change in vitiligo area scoring index (VASI), which was assessed on weeks 4, 8, 12 and 16. The secondary outcome measures included the patient's global assessment on VAS and investigator's global assessment based on photographic evaluation at baseline and after the treatment. Safety parameters included hemogram, LFTs, RFTs, CXR, ECG, urine, and stool examinations, which were evaluated at baseline and after the treatment. RESULTS: The per-protocol analysis was done on 30 participants in each group and the response in Unani group was not inferior to those receiving control group. The mean ± SD of vitiligo area scoring index (VASI) decreased from 4.09 ± 2.87 and 5.50 ± 5.73 at baseline to 3.13 ± 2.20 and 4.29 ± 4.95 at the end of the trial in both the Unani and control groups respectively. CONCLUSIONS: The study inferred that both the interventions are equally effective and well-tolerated in patients with non-segmental vitiligo.

The VEGAS Platform Is Unsuitable for Mammalian Directed Evolution.

Directed evolution uses cycles of gene diversification and selection to generate proteins with novel properties. While traditionally directed evolution is performed in prokaryotic systems, recently a mammalian directed evolution system (viral evolution of genetically actuating sequences, or "VEGAS") has been described. Here we report that the VEGAS system has major limitations that preclude its use for directed evolution. The deconstructed Sindbis virus (SINV) genome that comprises the VEGAS system could no longer promote Sindbis structural gene (SSG)-dependent viral replication. Moreover, viral particles generated using the VEGAS system rapidly lost the target directed evolution transgene, and instead, "cheater" particles, primarily containing RNA encoding SINV structural components, arose. By sequencing, we found that this contamination came from RNA provided during initial SINV packaging, not RNA derived from the VEGAS system. Of note, both the structural RNA and target transgenes used in the VEGAS system contain viral packaging sequences. The impact of SINV "cheater" particles could be potentially overcome in the context of a robust VEGAS circuit, but since SSG complementation is also defective in the VEGAS system, selection for authentic evolution products is not currently possible. Similar results have been obtained in independent laboratories. Taken together, these results show that the VEGAS system does not work as described and, without significant redesign, cannot be used for mammalian directed evolution campaigns.

Molecular, Biochemical, and Clinical Characterization of Thirteen Patients with Glycogen Storage Disease 1a in Malaysia.

Background: Glycogen storage disease type 1a (GSD1a) is a rare autosomal recessive metabolic disorder characterized by hypoglycaemia, growth retardation, lactic acidosis, hepatomegaly, hyperlipidemia, and nephromegaly. GSD1a is caused by a mutation in the G6PC gene encoding glucose-6-phosphatase (G6Pase); an enzyme that catalyses the hydrolysis of glucose-6-phosphate (G6P) to phosphate and glucose. Objective: To elaborate on the clinical findings, biochemical data, molecular genetic analysis, and short-term prognosis of 13 GSD1a patients in Malaysia. Methods: The information about 13 clinically classified GSD1a patients was retrospectively studied. The G6PC mutation analysis was performed by PCR-DNA sequencing. Results: Patients were presented with hepatomegaly (92%), hypoglycaemia (38%), poor weight gain (23%), and short stature (15%). Mutation analysis revealed nine heterozygous mutations; eight previously reported mutations (c.155 A > T, c.209 G > A, c.226 A > T, c.248 G > A, c.648 G > T, c.706 T > A, c.1022 T > A, c.262delG) and a novel mutation (c.325 T > C). The most common mutation found in Malaysian patients was c.648 G > T in ten patients (77%) of mostly Malay ethnicity, followed by c.248 G > A in 4 patients of Chinese ethnicity (30%). A novel missense mutation (c.325 T > C) was predicted to be disease-causing by various in silico software. Conclusions: The establishment of G6PC molecular genetic testing will enable the detection of presymptomatic patients, assisting in genetic counselling while avoiding the invasive methods of liver biopsy.

Recent Techniques in Determining the Effects of Climate Change on Depressive Patients: A Systematic Review.

Climate change is amongst the most serious issues nowadays. Climate change has become a concern for the scientific community as it could affect human health. Researchers have found that climate change potentially impacts human mental health, especially among depressive patients. However, the relationship is still unclear and needs further investigation. The purpose of this systematic review is to systematically evaluate the evidence of the association between climate change effects on depressive patients, investigate the effects of environmental exposure related to climate change on mental health outcomes for depressive patients, analyze the current technique used to determine the relationship, and provide the guidance for future research. Articles were identified by searching specified keywords in six electronic databases (Google Scholar, PubMed, Scopus, Springer, ScienceDirect, and IEEE Digital Library) from 2012 until 2021. Initially, 1823 articles were assessed based on inclusion criteria. After being analyzed, only 15 studies fit the eligibility criteria. The result from included studies showed that there appears to be strong evidence of the association of environmental exposure related to climate change in depressive patients. Temperature and air pollution are consistently associated with increased hospital admission of depressive patients; age and gender became the most frequently considered vulnerability factors. However, the current evidence is limited, and the output finding between each study is still varied and does not achieve a reasonable and mature conclusion regarding the relationship between the variables. Therefore, more evidence is needed in this domain study. Some variables might have complex patterns, and hard to identify the relationship. Thus, technique used to analyze the relationship should be strengthened to identify the relevant relationship.

Improvements of Flexural Properties and Thermal Performance in Thin Geopolymer Based on Fly Ash and Ladle Furnace Slag Using Borax Decahydrates.

This paper elucidates the influence of borax decahydrate addition on the flexural and thermal properties of 10 mm thin fly ash/ladle furnace slag (FAS) geopolymers. The borax decahydrate (2, 4, 6, and 8 wt.%) was incorporated to produce FAB geopolymers. Heat treatment was applied with temperature ranges of 300 °C, 600 °C, 900 °C, 1000 °C and 1100 °C. Unexposed FAB geopolymers experienced a drop in strength due to a looser matrix with higher porosity. However, borax decahydrate inclusion significantly enhanced the flexural performance of thin geopolymers after heating. FAB2 and FAB8 geopolymers reported higher flexural strength of 26.5 MPa and 47.8 MPa, respectively, at 1000 °C as compared to FAS geopolymers (24.1 MPa at 1100 °C). The molten B2O3 provided an adhesive medium to assemble the aluminosilicates, improving the interparticle connectivity which led to a drastic strength increment. Moreover, the borax addition reduced the glass transition temperature, forming more refractory crystalline phases at lower temperatures. This induced a significant strength increment in FAB geopolymers with a factor of 3.6 for FAB8 at 900 °C, and 4.0 factor for FAB2 at 1000 °C, respectively. Comparatively, FAS geopolymers only achieved 3.1 factor in strength increment at 1100 °C. This proved that borax decahydrate could be utilized in the high strength development of thin geopolymers.

The efficacy of topical Marham-e-Akbar in chronic atopic dermatitis - an open-label interventional study.

OBJECTIVES: Chronic atopic dermatitis (AD) is an inflammatory skin condition marked by intense pruritus, dry skin, and severe impact on the life quality of the patients. Conventionally, it is managed by using emollients, calcineurin inhibitors, and topical corticosteroids. In Unani medicine, eminent scholars advocated many drug formulations including topical Marham-e-Akbar for effective healing of AD but scientific evidence is scarce. Hence, this study was designed. METHODS: This was a single-arm clinical trial conducted on 30 participants aged 18-65 years suffering from chronic AD after obtaining written informed consent. The trial intervention was Marham-e-Akbar consisting of Murdar Sang (Plumbi oxidum); Sindur (red lead); olive oil (Olea europaea oil); Kath (Acacia catechu extract); Safeda Kashgari (Zinc oxide); Sirka (vinegar); and Phitkiri (alum) to be applied twice daily for 42 days. The objective parameters were SCORAD and DLQI, while the subjective parameters included itching, scaling, and erythema assessed on a customized VAS scale and 4-point Likert scale. RESULTS: The pre-post analysis inferred statistically significant attenuation in subjective parameters (itching, scaling, and erythema) and objective scales (SCORAD) and (DLQI) with p<0.001. CONCLUSIONS: The study findings deduced that Marham-e-Akbar is effective in the amelioration of chronic atopic dermatitis and quality of life of the patients as well.

CRISPR/Cas-Mediated Knock-in of Genetically Encoded Fluorescent Biosensors into the AAVS1 Locus of Human-Induced Pluripotent Stem Cells.

Genetically encoded fluorescent biosensors (GEFBs) enable researchers to visualize and quantify cellular processes in live cells. Induced pluripotent stem cells (iPSCs) can be genetically engineered to express GEFBs via integration into the Adeno-Associated Virus Integration Site 1 (AAVS1) safe harbor locus. This can be achieved using CRISPR/Cas ribonucleoprotein targeting to cause a double-strand break at the AAVS1 locus, which subsequently undergoes homology-directed repair (HDR) in the presence of a donor plasmid containing the GEFB sequence. We describe an optimized protocol for CRISPR/Cas-mediated knock-in of GEFBs into the AAVS1 locus of human iPSCs that allows puromycin selection and which exhibits negligible off-target editing. The resulting iPSC lines can be differentiated into cells of different lineages while retaining expression of the GEFB, enabling live-cell interrogation of cell pathway activities across a diversity of disease models.

Effect of structural variation on enzymatic activity in tetranuclear (Cu4) clusters with defective cubane core.

The stimulus to the modeling of enzyme functioning sites comes from their potential to give insight into the natural enzyme's mechanistic pathways, ascertain the role of that different metal ion in the active site and construct better catalysts motivated by nature. The presence of metal ion leads to the activation of molecular oxygen in the metalloenzymes. The metalloenzymes such as the catechol oxidase (CO) enzyme that oxidizes the catechol to corresponding quinones which eventually protect damage tissues from plant and pathogen. Thus, the design and characterization of catalysts used as selectively and efficiently oxidation reactions have grown to be unique challenges for modern inorganic chemists. In this work, two novel tetranuclear complexes (1 and 2) have been synthesized in excellent yield. The complexes were characterized using various spectroscopic techniques such as FTIR, UV-Visible and PXRD pattern. The structure of 1 and 2 was elucidated by SC-XRD (single crystal X-ray diffraction) analysis. The magnetic study reveals the presence of the antiferromagnetic nature of 1 and 2. Both 1 and 2 shows a very good catecholase-like activity by oxidizing the catechol to analogous quinone in methanolic solution. Thus, a structure-activity relationship can further help us design other substituted tetranuclear complexes with enhanced catecholase like activity.Communicated by Ramaswamy H. Sarma.

Elevated-Temperature Performance, Combustibility and Fire Propagation Index of Fly Ash-Metakaolin Blend Geopolymers with Addition of Monoaluminium Phosphate (MAP) and Aluminum Dihydrogen Triphosphate (ATP).

Thermal performance, combustibility, and fire propagation of fly ash-metakaolin (FA-MK) blended geopolymer with the addition of aluminum triphosphate, ATP (Al(H2PO4)3), and monoaluminium phosphate, MAP (AlPO4) were evaluated in this paper. To prepare the geopolymer mix, fly ash and metakaolin with a ratio of 1:1 were added with ATP and MAP in a range of 0-3% by weight. The fire/heat resistance was evaluated by comparing the residual compressive strengths after the elevated temperature exposure. Besides, combustibility and fire propagation tests were conducted to examine the thermal performance and the applicability of the geopolymers as passive fire protection. Experimental results revealed that the blended geopolymers with 1 wt.% of ATP and MAP exhibited higher compressive strength and denser geopolymer matrix than control geopolymers. The effect of ATP and MAP addition was more obvious in unheated geopolymer and little improvement was observed for geopolymer subjected to elevated temperature. ATP and MAP at 3 wt.% did not help in enhancing the elevated-temperature performance of blended geopolymers. Even so, all blended geopolymers, regardless of the addition of ATP and MAP, were regarded as the noncombustible materials with negligible (0-0.1) fire propagation index.

Capturing potential impact of challenge-based gamification on gamified quizzing in the classroom.

Challenges in education have continuously been addressed by integrating gamification, but a gap remains for game design principles that support user engagement. This paper outlines results obtained from integrating challenge-based gamification into an elementary school classroom to examine the emergence of student engagement and learning-related behavior. The approach was applied to logical puzzle quizzes where different gamification adjustments were captured and examined using physics' analogy (called the motion in mind concept). The structural experiment, with a mixed methods design, was designed around the notion of time pressure and the difficulty of gamifying the quizzing experience. This model was constructed to validate and expand the quantitative findings (motion in mind model) by including qualitative explorations (thematic analysis). The results revealed the potential synthesis of motion in mind and flow theory, and its relationships to engagement and learning were identified as a new conceptual scheme.