Microfluidics-based EGFR mutation detection and its implication in the resource-limited clinical setting.

Management of lung cancer today obligates a mutational analysis of the epidermal growth factor receptor (EGFR) gene particularly when Tyrosine Kinase Inhibitor (TKI) therapy is being considered as part of prognostic stratification. This study evaluates the performance of automated microfluidics-based EGFR mutation detection and its significance in clinical diagnostic settings. Formalin-fixed, paraffin-embedded (FFPE) samples from NSCLC patients (n = 174) were included in a two-phase study. Phase I: Validation of the platform by comparing the results with conventional real-time PCR and next-generation sequencing (NGS) platform. Phase II: EGFR mutation detection on microfluidics-based platform as part of routine diagnostics workup. The microfluidics-based platform demonstrates 96.5% and 89.2% concordance with conventional real-time PCR and NGS, respectively. The system efficiently detects mutations across the EGFR gene with 88.23% sensitivity and 100% specificity. Out of 144 samples analysed in phase II, the platform generated valid results in 94% with mutation detected in 41% of samples. This microfluidics-based platform can detect as low as 5% mutant allele fractions from the FFPE samples. Therefore the microfluidics-based platform is a rapid, complete walkaway, with minimum tissue requirement (two sections of 5 µ thickness) and technical skill requirement. The method can detect clinically actionable EGFR mutations efficiently and can be considered a reliable diagnostic platform in resource-limited settings. From receiving samples to reporting the results this platform provides accurate data without much manual intervention. The study helped to devise an algorithm that emphasizes effective screening of the NSCLC cases for EGFR mutations with varying tumour content. Thus it helps in triaging the cases judiciously before proceeding with multigene testing.

A Potpourri of Trichoscopic Findings: Unique Presentation of Hyperthyroidism-Induced Hair Changes.

Hyperthyroidism is known to cause thin, brittle hair, and premature graying. Hair shaft abnormalities in hyperthyroidism have not been described in the literature. A visible and microscopic change in the hair shaft due to cosmetic procedures and environmental factors is known as hair weathering. Herein, we report a case of hair weathering in an adult female with hyperthyroidism with unique dermoscopic features.

Evaluation of the Efficacy and Safety of Apremilast in the Management of Lichen Planus.

Introduction: Lichen planus is a chronic disease with often disappointing and less than optimal treatment options. Apremilast modulates inflammatory signalling pathways which play a central role in the pathogenesis of lichen planus, thus making it useful in the management of such patients. Materials and Methods: The present study was an investigator-initiated, single-centre, non-randomized, open-label, pilot study of the efficacy and safety of Apremilast in the treatment of lichen planus. All the patients were prescribed Apremilast 30mg, twice daily, for 12 weeks. Patients were evaluated for improvement in their lesions, based on the physician's global assessment (PGA), subject global assessment (SGA), and target area lesion symptom score (TALSS). Results: A total of 34 patients were included in the study; 26 patients completed the study duration and were considered for the final analysis. After 12 weeks, 34.61% (n = 9/26) patients showed 2 or more grade improvement in their disease as per PGA. About 42.30% (n = 11/26) patients achieved more than 50% improvement in their lesions based on the subject global assessment of their disease. There was a significant improvement in TALSS during the study period (p < 0.0001). Only 23.07% (n = 6/26) patients developed one or more adverse events because of Apremilast with headache being the commonest side effect. Conclusion: The results obtained in our study justify that Apremilast is efficacious and safe in the management of patients with lichen planus. Based on these results, Apremilast can be considered as a promising alternative treatment option in patients with lichen planus.

Comparative Assessment of DNA Extraction Techniques From Formalin-Fixed, Paraffin-Embedded Tumor Specimens and Their Impact on Downstream Analysis.

OBJECTIVES: Good-quality nucleic acid extraction from formalin-fixed, paraffin-embedded (FFPE) specimens remains a challenge in molecular-oncopathology practice. This study evaluates the efficacy of an in-house developed FFPE extraction buffer compared with other commercially available kits. METHODS: Eighty FFPE specimens processed in different surgical pathology laboratories formed the study sample. DNA extraction was performed using three commercial kits and the in-house developed FFPE extraction buffer. DNA yield was quantified by a NanoDrop spectrophotometer and Qubit Fluorometer, and its purity was measured by the 260/280-nm ratio. A fragment analyzer system was used for accurate sizing of DNA fragments of FFPE DNA. The downstream effects of all extraction methods were evaluated by polymerase chain reaction (PCR) and Sanger sequencing. RESULTS: In comparison with the commercial kits, the in-house buffer yielded higher DNA quantity and quality number (P < .0001). In addition, DNA integrity and fragment size were preserved in a significantly greater number of samples isolated with the in-house buffer (P < .05). The target PCR amplification rate with the in-house buffer extracted samples was also significantly higher, with 98% of the samples showing interpretable sequencing results. CONCLUSIONS: The in-house developed FFPE extraction buffer performed superior to other methods in terms of suitability for downstream applications, time, cost-efficiency, and ease of performance.

Impact of COVID-19 on quality checks of solid tumor molecular diagnostic testing-A surveillance by EQAS provider in India.

BACKGROUND: Molecular tests in solid tumours for targeted therapies call for the need to ensure precision testing. To accomplish this participation in the External Quality Assessment Program (EQAS) is required. This evaluates the consistency of diagnostic testing procedures and offers guidance for improving quality. Outbreak of COVID-19 pandemic led to worldwide lockdown and disruption of healthcare services including participation in EQAS.The present study describes the extended scope of EQAS offered byMPQAP (Molecular Pathology Quality Assurance Program), the first proficiency test provider for solid tumor diagnostics in India. The study surveys the preparedness of molecular testing laboratories in routine diagnostics and participation for quality assessment scheme. METHODS: A documented guideline for measures and precautions to be carried by testing laboratories in performing routine diagnostic tests during the lockdown period were charted and distributed to all MPQAP participant centres. A survey was conducted for MPQAP participants to check whether laboratories were involved in COVID-19 testing and to evaluate the impact of lockdown on the operations of diagnostics procedures. From the acquired response of the survey, 2 cycles out of initially proposed 11 cycles were executed with transformed approach using digital tools and image interpretation modules. FINDINGS: Out of 25 solid tumour testing laboratories registered as participants, 15 consented to participate in survey. The summary of survey conveyed the impact of COVID-19onroutine operations of diagnostics tests such as shortcomings in inventory and human resource management. Thirteen participants showed active willingness and consented to participate in EQAS test scheme. INTERPRETATIONS: The survey findings and assessment of EQAS cycles endorsed the quality testing procedures carried by participating laboratories throughout the lockdown. It highlighted the utility of EQAS participation during pandemic along with emphasis on safety measures for continual improvement in quality of diagnostic services.

An Open-Label, Randomized, Prospective, Comparative, Three-Arm Clinical Trial to Evaluate the Safety and Effectiveness of Apremilast with Three Different Titration Methods in Patients with Chronic Plaque Psoriasis in India.

Purpose: To minimize adverse effects (AEs), apremilast is recommended to titrate at the initiation of therapy. But still, many patients experience AEs, resulting in discontinuation of therapy. As a result, many dermatologists have adapted to further titrate apremilast in different ways. The present study was planned to evaluate the safety and effectiveness of apremilast in different dose titration methods as initiation therapy in the treatment of plaque psoriasis. Patients and Methods: In this open-label, randomized, prospective, comparative, three-arm, single center study, 128 plaque psoriasis patients were included. Patients were randomized into three groups. Group I received standard titration for the first 6 days; Group II received all tablets in a starter pack as once a day (OD) total for 13 days; and Group III received two starter packs as 8 tablets each of apremilast 10 mg and 20 mg as OD and 10 tablets of 30 mg as OD, in total for 26 days. All groups received apremilast 30 mg as twice a day after initial titration. The total duration of apremilast therapy in all groups was 16 weeks. Results: In safety assessment, AEs were reported in 50%, 41.3% and 25% in Groups I, II and III, respectively (p <0.05) with nausea being the most common AE. In Group I, 10.53% of patients discontinued apremilast whereas 6.52% and 2.27% discontinued in Groups II and III respectively. Maximum number of AEs were seen in Group I in first week only (74.19%) compared with other groups. At week 16, on the Psoriasis Area and Severity Index, PASI 75 was achieved in 31.43%, 42.4% and 33.3% of patients in Groups I, II and III, respectively with no statistical difference between any groups. Conclusion: It can be concluded that slower titration is a useful strategy for minimizing AEs while at the same time maintaining effectiveness of apremilast.

Estimation of Kidney Size From Foot Length in Newborns: A Cross-Sectional Study.

Background Kidney size determination and sonographic follow-up are important in clinical diagnosis and treatment in children. Various anthropometric measurements are correlated with gestational age and birth weight and are used to identify high-risk babies in need of early interventions. Although foot length has emerged as a simple and reliable anthropometric measurement, it is not correlated with kidney size, except in the fetal period. This study was undertaken to find a correlation between foot length and kidney dimensions and estimate kidney size by finding regression equations in newborns. Methods We conducted a cross-sectional study and 216 newborns were enrolled at a tertiary care hospital. Foot length was measured by digital Vernier calipers and kidney dimensions were measured by ultrasonography. The Pearson correlation coefficient and simple linear regression tests were used to determine the relationship between foot length and kidney dimensions. Results Foot lengths and kidney dimensions were comparable in males and females as well as on the right and left sides, except for kidney length, which was found to be longer in males. Both right and left foot lengths showed highly significant (p<0.001) but small, positive correlations with corresponding side kidney length, breadth, and area, with R-values ranging from 0.2874 to 0.3668. However, the correlation between birth weight and foot length was significant, positive, and moderate (r=0.6962 and 0.6923 for right and left foot lengths, respectively). The regression equation for estimation of kidney size from foot length was obtained but the variance explained was small (e.g. R2=0.1325 for right kidney length). Out of 216 babies in our study, 10 babies had a renal anomaly. Conclusions We found a significant but small, positive correlation between foot length and kidney dimensions. Only 13.25% of the variance in kidney length was associated with foot length. Birth weight also had a significant and positive but small correlation with kidney dimensions. However, the correlation of birth weight with foot length was moderate, and a 57.14% variance in foot length was associated with birth weight. Multivariate regression analysis with more anthropometric parameters and gestational age may help in finding a better estimation of kidney size.

A comparative study of propofol, thiopentone sodium, and ketofol as induction agents for electro convulsive therapy.

Background and Aims: Thiopentone and propofol are most commonly used induction agents for electro convulsive therapy (ECT). Recently, ketofol, an admixture of propofol and ketamine, is being tried in ECT. We aimed to compare propofol, thiopentone, and ketofol as induction agents during ECT regarding their effects on ECT-induced hemodynamic changes, seizure duration and recovery parameters. Material and Methods: This prospective randomized double blind study was conducted in 30 patients between 18 and 65 years with ASA status I and II scheduled for ECT. All patients received all study agents for first three sessions of ECT. The observations were compiled as Group K (Inj. Ketofol i.e., Inj. propofol 0.5mg/kg % Inj. ketamine 0.5mg/kg), Group P (Inj. propofol 1mg/kg), and Group T (Inj. thiopentone 3mg/kg). Heart rate (HR) and blood pressure (systolic, diastolic, and mean) was recorded at pre op, 0, 5, 10, and 20 min after ECT. The seizure duration, time to spontaneous eye opening, andobeying verbal commands and agitation score were recorded. Results: Statistically significant difference was seen in HR at 10 min after delivery of shock; in systolic BP at 2 min after shock; in diastolic BP after administration of study drug and immediately after shock and in mean arterial pressure at post induction, 0and2 min after shock with group T showing higher values compared to group K and P (p < 0.05). At all other times HR andBP was comparable in all the three groups. Seizure duration was more in group T than Groups P and K although the difference was statistically insignificant. Time to spontaneous eye opening and obey verbal commands was comparable in all groups. Mean agitation score was highest in group T than Groups P& Kwith Group P showing least value (p = 0.003). Conclusion: Propofol and ketofol showed superior hemodynamic stability than thiopentone but comparable seizure duration and recovery parameters. Thus, propofol and ketofol can be effectively used as induction agents for ECT although propofol is associated with lesser agitation than ketofol.

Contrary to Myth, Older Adults Multitask With Media and Technologies, But Studying Their Multitasking Behaviors Can Be Challenging.

BACKGROUND AND OBJECTIVES: The study's objective was to explore older adults' (aged 65 or older) descriptions of behavior related to multitasking with traditional and newer media/information and communication technologies (ICTs) and perceived benefits of such behavior, along with older adults' preference for research methods used to study their multitasking behaviors. Employing common media-use measures that heavily rely on self-reporting in populations of older adults is challenging, especially given that patterns of media/ICT use are becoming increasingly complex. Cumulatively, people spend more time using media than they are aware of because of the tendency to use some forms of media simultaneously. As cognitive ability deteriorates with age, self-reported recollection of complex patterns of media/ICT use, such as multitasking, among older adults increases the threat to data accuracy.Research Design and Methods: Twenty-eight community-dwelling older adults in a Midwestern U.S. state participated in in-depth interviews (average length was 40 minutes) to discuss their use of traditional and newer media/technologies in combination with other activities and outline methods researchers should use to study such behaviors. RESULTS: Participants reported they engaged in multitasking behaviors similar to those of younger generations, with the difference in the higher extent of using traditional media and ICTs. They talked about multitasking with radio and television for "background noise" as being a rewarding experience. They perceived the effects of multitasking to be detrimental to attention and performance and attributed this media-use habit to individual psychological and demographic differences. They preferred ethnographic observation and keeping a paper-and-pencil diary as research methods to study multitasking among their peers. Data-logging methods were less popular because they raised privacy concerns among interviewees. DISCUSSION AND IMPLICATIONS: Different types of traditional and newer media and technologies could be used differently in situations that require older adults to focus, relax, or be efficient. The findings suggest that future researchers strive for a compromise between data access and data accuracy when they select a research method to study media use among older adults.

Liquid Drug Dosage Measurement Errors with Different Dosing Devices.

An observational study was carried out to determine the magnitude of dosing errors made by parents, the most-preferred drug delivery device and the association of age, gender, education of the caregiver and number of children with the proportion of accurate doses. After enrolment, parents of children aged 6-60 mo were instructed to measure 5 ml of syrup paracetamol using any of the devices (stainless steel spoon, disposable plastic syringe, dosing cup with etched markings) displayed. The quantum of measured dose was confirmed using a calibrated glass cylinder. Error was defined as over 10% variation around the prescribed dose. Of 386 participants, 72 (18.65%) committed error, with 58 (15.02%) and 14 (3.62%) committing mild and moderate errors, respectively. Measuring cup (270, 69.95%) was the commonest device chosen. Use of syringe was associated with greater accurate measurements (P < 0.05) with only 3 (3.57%) committing error compared to 18 (56.25%) and 51 (18.88%) committing error with spoon and cup, respectively. On multivariate analysis, device was the only factor significantly associated with accuracy in measurements.

A Swelling over Sternum in a Child: Reminder of an Uncommon Diagnosis.

Lack of awareness about isolated tubercular osteomyelitis of the sternum resulted in a delay in diagnosing the condition in an eleven-year-old girl who presented with a gradually increasing swelling over the sternum. Radiological, histological, and microbiological investigations helped diagnose the condition and the child responded well to antitubercular therapy (ATT) and surgical debridement. The report provides a brief description about the various management options available.

Air-lift reactor system for the treatment of waste-gas-containing monochlorobenzene.

An air-lift bioreactor (ALR) system, applied for the treatment of waste-gas-containing monochlorobenzene (MCB) was seeded with pure culture of Acinetobacter calcoaceticus, isolated from soil as a starter seed. It was found that MCB was biologically converted to chloride as chloride was mineralized in the ALR. After the built up of the biomass in the ALR, the reactor parameters which have major influence on the removal efficiency and elimination capacity were studied using response surface methodology. The data generated by running the reactor for 150 days at varying conditions were fed to the model with a target to obtain the removal efficiency above 95% and the elimination capacity greater than 60%. The data analysis indicated that inlet loading was the major parameter affecting the elimination capacity and removal efficiency of >95%. The reactor when operated at optimized conditions resulted in enhanced performance of the reactor.