Stability and dynamics of extradenticle modulates its function.

Extradenticle (EXD) is a partner protein of the HOX transcription factors and plays an important role in the development of Drosophila. It confers increased affinity and specificity of DNA-binding to the HOX proteins. However, the DNA-binding homeodomain of EXD has a significantly weaker affinity to DNA compared to the HOX homeodomains. Here, we show that a glycine residue (G290) in the middle of the EXD DNA-binding helix primarily results in this weaker binding. Glycine destabilizes helices. To probe its role in the stability and function of the protein, G290 was mutated to alanine. The intrinsic stability of the DNA-binding helix increased in the G290A mutant as observed by NMR studies and molecular dynamics (MD) simulation. Also, NMR dynamics and MD simulation show that dynamic motions present in the wild-type protein are quenched in the mutant. This in turn resulted in increased stability of the entire homeodomain (ΔΔGG→A of -2.6 kcal/mol). Increased protein stability resulted in three-fold better DNA-binding affinity of the mutant as compared to the wild-type protein. Molecular mechanics with generalized Born and surface area solvation (MMGBSA) analysis of our MD simulation on DNA-bound models of both wild-type and mutant proteins shows that the contribution to binding is enhanced for most of the interface residues in the mutant compared to the wild-type. Interestingly, the flexible N-terminal arm makes more stable contact with the DNA minor groove in the mutant. We found that the two interaction sites i.e. the DNA-binding helix and the unstructured N-terminal arm influence each other via the bound DNA. These results provide an interesting conundrum: alanine at position 290 enhances both the stability and the DNA-binding affinity of the protein, however, evolution prefers glycine at this position. We have provided several plausible explanations for this apparent conundrum. The function of the EXD as a HOX co-factor requires its ability to discriminate similar DNA sequences, which is most likely comprom.

Experimental methods to study the structure and dynamics of intrinsically disordered regions in proteins.

Eukaryotic proteins often feature long stretches of amino acids that lack a well-defined three-dimensional structure and are referred to as intrinsically disordered proteins (IDPs) or regions (IDRs). Although these proteins challenge conventional structure-function paradigms, they play vital roles in cellular processes. Recent progress in experimental techniques, such as NMR spectroscopy, single molecule FRET, high speed AFM and SAXS, have provided valuable insights into the biophysical basis of IDP function. This review discusses the advancements made in these techniques particularly for the study of disordered regions in proteins. In NMR spectroscopy new strategies such as 13C detection, non-uniform sampling, segmental isotope labeling, and rapid data acquisition methods address the challenges posed by spectral overcrowding and low stability of IDPs. The importance of various NMR parameters, including chemical shifts, hydrogen exchange rates, and relaxation measurements, to reveal transient secondary structures within IDRs and IDPs are presented. Given the high flexibility of IDPs, the review outlines NMR methods for assessing their dynamics at both fast (ps-ns) and slow (µs-ms) timescales. IDPs exert their functions through interactions with other molecules such as proteins, DNA, or RNA. NMR-based titration experiments yield insights into the thermodynamics and kinetics of these interactions. Detailed study of IDPs requires multiple experimental techniques, and thus, several methods are described for studying disordered proteins, highlighting their respective advantages and limitations. The potential for integrating these complementary techniques, each offering unique perspectives, is explored to achieve a comprehensive understanding of IDPs.

Real world outcome of B ALL with t (1; 19) (q23; p13)/TCF3::PBX1 in adolescents and adults treated with intensive regimes.

Significant heterogeneity has been reported in outcome of Acute lymphoblastic leukemia with t(1;19)(q23;p13)/TCF3::PBX1 in adolescents and adults leading to a lack of consensus on precise risk stratification. We evaluated clinical outcome of 17 adult ALL cases (≥15 years) with this genotype treated on intensive regimes.13/17 received COG0232 and 4/17 cases received UK-ALL protocol. All achieved CR (100%) with above treatment. End of induction MRD was evaluated in 14/17 cases of which 11 (78.5%) achieved MRD negativity. Total nine patients relapsed (7 marrows, 2 CNS). Overall survival at 2 years was 53.3%. The 2 year estimated PFS was 42.9%. The 2 years CIR was 54.2%. Adults with this genotype perform poorly despite early favorable response. Incorporation of novel immunotherapies and prompt HSCT should be strongly considered with this genotype. Targeted NGS panels for additional genetic aberrations can further help in risk stratifying and guiding therapy for this genotype.

Social media use among adolescents with eating disorders: a double-edged sword.

Eating disorders are on the rise with a significant impact on mental health. Misuse of social media platforms is likely a significant contributing factor. This trend is especially pronounced among adolescents, who are increasingly using social media platforms for communications, building relationships, learning and entertainment. The unsupervised sharing of social media content can have drastic consequences on the physical and psychological wellbeing of youth, which often takes the form of "thinspiration" or "fitspiration". This probably serves as a trigger for those already struggling with eating disorders. In addition, there is a lack of awareness among youth and adults on ideal knowledge-sharing practices related to an eating disorder. By addressing the unique challenges that social media presents for youth with eating disorders, communities can work towards creating a safer and more supportive online environment on a global scale. At this junction, this article aims to share the current challenges of social media use among adolescents with eating disorders and put forth recommendations for how social media could be used as a tool for positive impact in this population.

Customizing Starch Properties: A Review of Starch Modifications and Their Applications.

Starch has been a convenient, economically important polymer with substantial applications in the food and processing industry. However, native starches present restricted applications, which hinder their industrial usage. Therefore, modification of starch is carried out to augment the positive characteristics and eliminate the limitations of the native starches. Modifications of starch can result in generating novel polymers with numerous functional and value-added properties that suit the needs of the industry. Here, we summarize the possible starch modifications in planta and outside the plant system (physical, chemical, and enzymatic) and their corresponding applications. In addition, this review will highlight the implications of each starch property adjustment.

Immunosuppression Boost With Mycophenolate Mofetil for Mixed Chimerism in Thalassemia Transplants.

Declining mixed chimerism (MC) portending impending graft failure is an undesirable outcome. However, for hemoglobinopathies in a stable state of MC, residual host cells persist without rejection in 30% to 40% of patients after hematopoietic stem cell transplantation (HSCT). Early detection and level of MC have been attributed to be significant in predicting the outcome of MC. Common clinical approach on MC is removal of immunosuppression. We retrospectively evaluated MC in transfusion dependent thalassemia patients who underwent HSCT in our institution between September 2013 and January 2022 to determine the outcome of MC on the basis of our approach of immunosuppression boost in comparison to conventional approach of immunosuppression tapering. Among 90 patients, 22 (24.4 %) had MC at some time point after transplantation with a median follow-up of 496 (67-1492) days. Immunosuppression withdrawal was done in 12 (54.5%) patients, whereas immunosuppression boost was given in 8 (36.3%) patients. In the immunosuppression withdrawal group, 2 (16.6%) patients evolved to complete chimerism, 5(41.6%) patients had persistent MC (PMC), whereas 5 (41.6%) patients had secondary rejection. All these rejections were at median of 186 (89-251) days after transplantation. In the immunosuppression boost group, all patients (n = 8) had PMC with no secondary rejection until median follow-up of 255(97-812) days after transplantation. We acknowledge that we need more experience with our unconventional approach of immunosuppression boost to obtain statistical significance in comparison to the conventional approach of tapering of immunosuppression.

An observational multi-centric COVID-19 sequelae study among health care workers.

Background: India has seen more than 43 million confirmed cases of COVID-19 as of April 2022, with a recovery rate of 98.8%, resulting in a large section of the population including the healthcare workers (HCWs), susceptible to develop post COVID sequelae. This study was carried out to assess the nature and prevalence of medical sequelae following COVID-19 infection, and risk factors, if any. Methods: This was an observational, multicenter cross-sectional study conducted at eight tertiary care centers. The consenting participants were HCWs between 12 and 52 weeks post discharge after COVID-19 infection. Data on demographics, medical history, clinical features of COVID-19 and various symptoms of COVID sequelae was collected through specific questionnaire. Finding: Mean age of the 679 eligible participants was 31.49 ± 9.54 years. The overall prevalence of COVID sequelae was 30.34%, with fatigue (11.5%) being the most common followed by insomnia (8.5%), difficulty in breathing during activity (6%) and pain in joints (5%). The odds of having any sequelae were significantly higher among participants who had moderate to severe COVID-19 (OR 6.51; 95% CI 3.46-12.23) and lower among males (OR 0.55; 95% CI 0.39-0.76). Besides these, other predictors for having sequelae were age (≥45 years), presence of any comorbidity (especially hypertension and asthma), category of HCW (non-doctors vs doctors) and hospitalisation due to COVID-19. Interpretation: Approximately one-third of the participants experienced COVID sequelae. Severity of COVID illness, female gender, advanced age, co-morbidity were significant risk factors for COVID sequelae. Funding: This work is a part of Indian Council for Medical Research (ICMR)- Rational Use of Medicines network. No additional financial support was received from ICMR to carry out the work, for study materials, medical writing, and APC.

Treatment of Ekbom Syndrome With Clozapine and Electroconvulsive Therapy.

Delusion of parasitosis (DP), which is also known as Ekbom syndrome, is a delusional disorder characterised by a false, fixed belief of being infested by insects or mites, despite the lack of supporting medical evidence. This disorder presents most commonly with the "Matchbox sign." DP can present as a primary or secondary delusional disorder. It can be associated with various psychiatric conditions such as schizophrenia spectrum disorders, mood disorders, anxiety or substance abuse. Several organic conditions such as dementia, malignancies, vitamin deficiencies and cerebrovascular accidents can mimic symptoms of DP. Hereby, we present a case of schizophrenia in a young woman associated with DP in our inpatient care and the treatment outcome with the use of clozapine and electroconvulsive therapy (ECT) regimen.

N-acetylcysteine in the Treatment of Internet Gaming Disorder.

Internet gaming disorder (IGD) is the persistent and recurrent use of the internet to engage in video gaming through a single or multiplayer interface that can lead to significant impairment or distress. With technological advancements in the last decade via portable handheld devices, along with their global availability, video games have found a new medium in which they can provide instantaneous access for casual and enthusiastic users alike. Unfortunately, this exponentially increases the possibility of addiction. IGD shares a similar pathophysiological etiology to addiction as drugs or gambling. However, it can be challenging to manage IGD due to the ease of video game access and limited understanding of the newly recognized disorder. This study aims to fill in the knowledge gap concerning the limited research on internet gaming addiction, its consequential effects on human cognitive-behavioral functioning, and pharmacotherapy management as observed in our patient, who developed IGD, starting initially as a casual recreational hobby among peers. This case also highlights the lack of social awareness and seriousness attributed to this disorder. It focuses on using N-acetylcysteine in the management as well as other psychological and psychotropic drugs.

Busulfan and cyclophosphamide-based conditioning regimen still holds the promise of being a safe and efficacious regimen for allogeneic transplantation in patients with transfusion-dependent thalassemia, even in high risk.

Busulfan and cyclophosphamide (BuCy)-based regimen has been used as a standard myeloablative chemotherapy for haematopoietic stem cell transplantation in thalassemia. However, treosulfan-based conditioning regimen has emerged due to concerns of toxicities. We retrospectively analysed the safety and efficacy of fludrabine/Bu/Cy/antithymocyte globulin (ATG) versus treosulfan/thiotepa/fludrabine regimens for Hematopoietic Stem Cell Transplant (HSCT) in transfusion-dependent thalassemia (TDT) conducted at our institute (2013-2021). In 75 patients, 36 (48%) received Flu/Bu/Cy/ATG whereas 39 (52%) received Treo/Thio/Flu. Median age was 6 (1-12) and 9 (1-15) years, respectively. Number of patients with Classes I, II, and III were 14, 10, and 12 in Flu/Bu/Cy/ATG versus 2, 19, and 18 in Treo/Thio/Flu group, respectively. Graft was growth factor mobilized bone marrow in Flu/Bu/Cy/ATG versus peripheral blood stem cell in Treo/Thio/Flu group. Mean stem cell dose was 3.82 (2.2-9.1) versus 5 (1.65-8.01) 106 /kg in Flu/Bu/Cy/ATG versus Treo/Thio/Flu group, respectively. Neutrophils and platelets engrafted at a median of 16 (14-21) and 16 (9-47) days in Flu/Bu/Cy/ATG and 15 (10-20) and 13 (9-41) days in Treo/Thio/Flu group. Median duration of follow-up was 28 (23-32.9) months. Five (6.6%) patients had rejection (all secondary). Venoocclusive disease was observed in 2 (5.7%) versus 4 (10.3%) patients (p = .047), respectively. Flu/Bu/Cy/ATG had 4 (11.4%) patients with acute GVHD versus 15 (38.5%) patients which had significant impact on survival (p = .038). We observed chronic GVHD in 4 (11.4%) and 11 (28.2%) patients, respectively, with significant impact on survival (p = .031). Four (5.1%) patients had TRM in Treo/Thio/Flu group, in contrast to none in Flu/Bu/Cy/ATG group. Mixed chimerism was common in Flu/Bu/Cy/ATG {20 (57.1%)} versus Treo/Thio/Flu group {12 (30.1%)}. Five-year Event Free Survival (EFS) and OS of entire cohort were 87% + 4% and 94% + 3%, respectively. Estimated TFS, EFS, OS of Flu/Bu/Cy/ATG versus Treo/Thio/Flu was 97.1% + 2.9% versus 89.2% + 5.1% (p = .251), 97 + 3% versus 80.7 + 6% (p = .041) and 100% versus 90.4 + 5% (p = .067), respectively. In our experience, Flu/Bu/Cy/ATG regimen is safe and effective even in high-risk TDT. However, one needs to be vigilant for mixed chimerism.

LIKE EARLY STARVATION 1 alters the glucan structures at the starch granule surface and thereby influences the action of both starch-synthesizing and starch-degrading enzymes.

For starch metabolism to take place correctly, various enzymes and proteins acting on the starch granule surface are crucial. Recently, two non-catalytic starch-binding proteins, pivotal for normal starch turnover in Arabidopsis leaves, namely, EARLY STARVATION 1 (ESV1) and its homolog LIKE EARLY STARVATION 1 (LESV), have been identified. Both share nearly 38% sequence homology. As ESV1 has been found to influence glucan phosphorylation via two starch-related dikinases, α-glucan, water dikinase (GWD) and phosphoglucan, water dikinase (PWD), through modulating the surface glucan structures of the starch granules and thus affecting starch degradation, we assess the impact of its homolog LESV on starch metabolism. Thus, the 65-kDa recombinant protein LESV and the 50-kDa ESV1 were analyzed regarding their influence on the action of GWD and PWD on the surface of the starch granules. We included starches from various sources and additionally assessed the effect of these non-enzymatic proteins on other starch-related enzymes, such as starch synthases (SSI and SSIII), starch phosphorylases (PHS1), isoamylase and ß-amylase. The data obtained indicate that starch phosphorylation, hydrolyses and synthesis were affected by LESV and ESV1. Furthermore, incubation with LESV and ESV1 together exerted an additive effect on starch phosphorylation. In addition, a stable alteration of the glucan structures at the starch granule surface following treatment with LESV and ESV1 was observed. Here, we discuss all the observed changes that point to modifications in the glucan structures at the surface of the native starch granules and present a model to explain the existing processes.

Psychiatric morbidity in absentees in industry.

BACKGROUND: Absenteeism refers to a worker's unscheduled absence from the workplace. Around the world, a large share of mental disorders can be attributed to work productivity losses. Despite industries working on improvement in employees' wages, better work environment and improved health care to improve their mental health. Absenteeism continues to be a global burden. AIM: The aim of the study was to explore the factors influencing absenteeism. MATERIALS AND METHODS: A sample size of 100 workers was selected and standardized, and reliable research tools were employed. Factors influencing absenteeism such as the sociodemographic factors, personality traits, psychiatric morbidity, family typology, and interactions were studied using relevant scales, i.e., Eysenck's Personality Inventory and Family Typology by Batti and Channabasavanna. RESULTS: Study found that absentees were more emotionally stable, whereas regulars were found to be emotionally unstable. Factors were found to be associated with absenteeism such as low income, 11-20 years of service, low mean neuroticism score, chronic physical disease, high psychiatric morbidity among the workers, egoistic and anomic family typology. This study reveals that a significantly higher proportion of absentees had a low pay scale. Paradoxically, a high proportion of absenteeism had a longer duration service, indicating that the low pay scales could have resulted from disciplinary actions against absentees, like loss of increments, etc. CONCLUSION: Psychosis, alcoholism, and chronic physical illnesses are among the illnesses which contribute to absenteeism.

Prevalence of suicidal ideation, plan, attempts and its associated factors in selected rural and urban areas of Puducherry, India.

BACKGROUND: The Union Territory of Puducherry has a high rate of suicide in India. This study aims to find the prevalence of suicidal ideation, plan and suicide attempts and the risk factors of suicidal behaviours. METHODS: This community-based cross-sectional study was conducted among adults aged 18 years and above in rural and urban areas of Puducherry, India. Columbia Suicide Severity Rating Scale questionnaire was used. Prevalence ratio was estimated to assess the factors associated. RESULTS: The 12-month and lifetime prevalence of suicidal ideation was reported to be 12.5% (95% CI: 10.4-15.0%) and 20% (95% CI: 17.4-22.9%), respectively. About 0.7% (95% CI: 0.3-1.5%) and 2.1% (95% CI: 1.2-3.3%) of the participants had suicide plan in past 12 months and ever in their lifetime, respectively. Around 1% (95% CI: 0.5-2.0%) and 5.7% (95% CI: 4.3-7.6%) of the participants had attempted suicide in past 12 months and ever in their lifetime, respectively. Family issues, physical abuse, unemployment and place of residence were significantly associated with suicidal ideation in multivariable analysis. CONCLUSION: Prevalence of suicidal ideation and suicide attempt were very high in the study area. Unemployment, physical abuse and family issues were significantly associated with suicidal behaviour.

Should Internet gaming disorder be considered a subtype of generalized problematic internet use? Findings from a study among medical college students.

PURPOSE: The current study aimed to explore whether Internet gaming disorder (IGD) should be considered a subtype of generalised problematic Internet use (PIU) by assessing the degree of overlap between them and comparing their correlates. The sociodemographic profile and presence of depressive symptoms were studied as correlates for IGD and generalised PIU. DESIGN AND METHODS: This was a self-report survey administered online among medical students (n = 306). FINDING: Less than half of the participants with IGD (45.45%; 5 out of 11) also met the criteria for generalised PIU. Only 6.67% (5 out of 75) of subjects with generalised PIU were also classified as having IGD. Male gender and time spent on digital gaming per day were associated with greater risk of IGD; whereas the time spent on Internet per day and more frequent use of the Internet for social networking rather than academic purposes were associated with greater risk of generalised PIU. The regression models built for IGD and generalised PIU were statistically significant (F = 64.10; P < .01 and F = 80.97; P < .01, respectively). The two models explained 66.1% and 57.4% of the variance in IGDS-SF and GPIUS2 scores, respectively. PRACTICE IMPLICATIONS: The IGD and generalised PIU appear to be conceptually distinct entities. The IGD should not be assumed as a subtype of generalised PIU. Thus, health care providers need to focus on IGD beyond and besides generalised PIU. There is a need to specifically enquire about IGD while screening for behavioural addictions involving use of Internet.

Physical fitness and its association with overweight among early adolescents in Puducherry, Southern India.

OBJECTIVES: Physical fitness is an important indicator of health and longevity. This can reduce morbidity and mortality of Non-communicable diseases. We aimed to assess the physical fitness among early adolescents and its association with age, gender and obesity. METHODS: We conducted a cross sectional analytical study in a public school in Puducherry, south India among early adolescents (aged 10-14 years). We assessed the physical fitness components like body composition (body mass index, waist circumference), muscular strength and endurance (hand grip test, curl up test), flexibility (sit and reach test, shoulder stretch test), agility (shuttle run test) and cardio respiratory and endurance (600-yard run test). RESULTS: A total of 263 children were included; 141 (54%) were girls. 30 children (11%) were thin and 57 (22%) were either obese or overweight. Overall, median (IQR) right- and left-hand grip strength was 10 (6-14) and 13 (7-18) unit of kg respectively. Median (IQR) curl-ups performed in a minute were 14 (10-20). About 13% of children failed to perform left shoulder stretch. Children aged 12-14 years had higher median (IQR) right hand grip strength [14.5 (12-22) unit of kg] compared to children aged 10 and 11 years [6 (4-12) unit of kg] and this difference was statistically significant (p<0.001). The performance was similar in all tests between boys and girls except for left hand grip strength; boys had higher grip strength compared to girls (p=0.02). In girls, children who belonged to thinner BMI category performed better than obese children in shuttle run test (p=0.03). CONCLUSION: There was a significant difference in physical fitness based on age except in left and right shoulder stretch test. Since the prevalence of overweight/obesity was high in adolescents, creating awareness and enabling environment related to the physical fitness is important.

Cytological diagnosis of juvenile xanthogranuloma: A rare histiocytic disorder.

Juvenile xanthogranuloma (JXG) is a rare type of non-Langerhan cell histiocytic disorder, which is mostly confined to skin of head and neck. It is a self-limiting benign condition, which does not require surgery. We present a case of 8-month-old girl child with multiple yellowish brown colored papules over scalp, face, and neck. A clinical diagnosis of cutaneous mastocytosis was made. Fine-needle aspiration cytology (FNAC) smears showed foamy macrophages along with mixed inflammatory infiltrate and few touton giant cells. A diagnosis of JXG was rendered which was confirmed on histopathology and immunohistochemistry.Juvenile xanthogranuloma can be diagnosed on FNAC based on its characteristic cytologic features; however, it requires a high index of suspicion by cytopathologist. Cytological diagnosis of JXG can save the patient from unnecessary surgical biopsy or excision.