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Plasma-derived extracellular vesicles (pEVs) are a potential source of diseased biomarker proteins. However, characterizing the pEV proteome is challenging due to its relatively low abundance and difficulties in enrichment. This study presents a streamlined workflow to identify EV proteins from cancer patient plasma using minimal sample input. Starting with 400 µL of plasma, we generated a comprehensive pEV proteome using size exclusion chromatography (SEC) combined with HiRIEF prefractionation-based mass spectrometry (MS). First, we compared the performance of HiRIEF and long gradient MS workflows using control pEVs, quantifying 2076 proteins with HiRIEF. In a proof-of-concept study, we applied SEC-HiRIEF-MS to a small cohort (12) of metastatic lung adenocarcinoma (LUAD) and malignant melanoma (MM) patients. We also analyzed plasma samples from the same patients to study the relationship between plasma and pEV proteomes. We identified and quantified 1583 proteins in cancer pEVs and 1468 proteins in plasma across all samples. While there was substantial overlap, the pEV proteome included several unique EV markers and cancer-related proteins. Differential analysis revealed 30 DEPs in LUAD vs the MM group, highlighting the potential of pEVs as biomarkers. This work demonstrates the utility of a prefractionation-based MS for comprehensive pEV proteomics and EV biomarker discovery. Data are available via ProteomeXchange with the identifiers PXD039338 and PXD038528.
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Artificial intelligence (AI) is increasingly being applied to wastewater treatment to enhance efficiency, improve processes, and optimize resource utilization. This review focuses on objectives, advantages, outputs, and major findings of various AI models in the three key aspects: the prediction of removal efficiency for both organic and inorganic pollutants, real-time monitoring of essential water quality parameters (such as pH, COD, BOD, turbidity, TDS, and conductivity), and fault detection in the processes and equipment integral to wastewater treatment. The prediction accuracy (R2 value) of AI technologies for pollutant removal has been reported to vary between 0.64 and 1.00. A critical aspect explored in this review is the cost-effectiveness of implementing AI systems in wastewater treatment. Numerous countries and municipalities are actively engaging in pilot projects and demonstrations to assess the feasibility and effectiveness of AI applications in wastewater treatment. Notably, the review highlights successful outcomes from these initiatives across diverse geographical contexts, showcasing the adaptability and positive impact of AI in revolutionizing wastewater treatment on a global scale. Further, insights on the ethical considerations and potential future directions for the use of AI in wastewater treatment plants have also been provided.
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Inteligência Artificial , Águas Residuárias , Eliminação de Resíduos Líquidos/métodos , Purificação da Água/métodosRESUMO
This case report explores a rare complication of broad ligament hematoma post-vaginal delivery, emphasizing the importance of prompt intervention in cases of postpartum hemorrhage with atypical presentations. A 22-year-old primigravida, at 39 weeks with hypothyroidism, presented with intermittent abdominal pain and normal fetal movements. After a normal vaginal delivery with a right mediolateral episiotomy, she developed intense perineal pain and hypotension due to a broad ligament hematoma. The surgical intervention included the evacuation of the hematoma, laparotomy, and internal iliac artery ligation. The postoperative care involved treatment for a methicillin-resistant Staphylococcus aureus (MRSA) infection, and the patient received blood transfusions. The follow-up showed complete wound healing and an uneventful postnatal period, with the patient resuming normal activities after three weeks. Comparison with similar cases in the literature highlighted various etiologies and clinical presentations of broad ligament hematoma, ranging from broad ligament pregnancy to uterine perforation. Timely surgical exploration, hematoma evacuation, and arterial ligation were essential in preventing adverse maternal outcomes, underscoring the importance of multidisciplinary collaboration and vigilant postoperative monitoring. The report emphasizes the need for a high index of suspicion and prompt intervention to ensure optimal recovery and minimize complications in cases of broad ligament hematoma following vaginal delivery.
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Trigonella, commonly known as Fenugreek, is among the most promising medicinal herbs consumed worldwide due its protein rich dietary contributions. This study involved induced mutagenesis on two Trigonella species (Trigonella foenum-graecum var. PEB and Trigonella corniculata var. Pusa kasuri) using caffeine and sodium azide as mutagens, resulting in the identification of nine high-yielding mutant lines in the M3 generation. Molecular characterization using SCoT markers revealed a high polymorphism of 28.3% and 46.7% in PEB and Pusa kasuri, respectively, facilitating the investigation of genetic divergence among the control and mutant lines. Similarity correlation analysis indicated a high similarity between mutant A and mutant C (0.97) and between mutant J and mutant O (0.88), while the lowest similarity was observed between mutant B and mutant F (0.74) and between control and mutant L (0.58). Mutant F and Mutant J displayed the highest seed yield and its attributing traits, and seed protein content in PEB and Pusa kasuri, respectively. Physiological parameters, including chlorophyll content (Mutants A and N) and carotenoids (mutant A and J), exhibited improvements. Assessment of stomatal and seed characteristics using scanning electron microscopy may lead to improved physiological processes and distinction at the interspecific level, respectively. Methanolic extracts of the control and the mutant lines of both species were subjected to GC-MS analysis, revealing 24 major phytocompounds known for their pharmacological activities (antioxidant, anti-inflammatory, anticancer, etc.). Statistical methods such as Pearson correlation heatmap and pairwise scatter plot matrix provided insights into the correlations and linear associations among parameters for both PEB and Pusa kasuri. The strong correlation between iron content and seeds per pod in the mutant lines suggests a promising avenue for further research. Continued research and breeding efforts using these mutants can lead to significant advancements in agriculture and medicine, benefiting farmers, consumers, and industries alike.
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Mutação , Sementes , Trigonella , Trigonella/genética , Trigonella/química , Trigonella/metabolismo , Sementes/genética , Sementes/metabolismo , Sementes/química , Mutagênese , Carotenoides/metabolismo , Cafeína/farmacologia , Clorofila/metabolismo , Azida Sódica/farmacologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Background: The accumulation of poorly folded protein in the endoplasmic reticulum (ER) promotes ER stress and contributes to the pathogenesis of hepatocellular carcinoma (HCC). Current therapies have various adverse effects, therefore, laying the need for an alternative approach. Kaempferol (KP), a naturally occurring flavonoid, possesses potent anti-proliferative properties against various cancer cells. Nevertheless, its involvement in HCC remains relatively unexplored, particularly regarding its influence on apoptosis and autophagy pathways. Methods: The effect of KP on cell viability, and motility of Hep3B cells was evaluated by MTT, and scratch assay, respectively. Hoechst staining and FACS analysis were done to check the effect of KP on apoptosis and cell cycle progression. qRTPCR was used to evaluate the expression of several apoptosis and autophagy-related genes. KP was docked with several ER stress-related proteins involved in HCC to gain further insights into molecular mechanisms. The results of docking studies were validated with MD simulation and in vitro studies. Results: Treatment with KP at different time intervals showed dose- and time-dependent growth inhibition of liver cancer cells. KP decreased motility and arrested the cell cycle at the G0/G1 phase in Hep3B cells. Additionally, in the context of HCC, the relationship between KP, apoptosis, and autophagy is significant. It induced apoptosis and autophagy in Hep3B cells by downregulating the expression of Bcl-2 and upregulated Bax and Bid, Caspase-3, Beclin-1, and LC3. KP showed a better binding affinity with Nrf2, PERK, and IRE1α among all selected proteins. Further, it reversed the protective effect of 4-PBA (ER Stress inhibitor) by inducing apoptosis and autophagy in Hep3B cells. Conclusion: The study suggested KP as a potential chemopreventive agent for managing HCC by effectively inducing apoptosis and autophagy in Hep3B cells.
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The auxin-inducible degron (AID) system is a powerful tool to deplete proteins in vivo. Here, we present a protocol for AID-mediated depletion of two proteins (CFI-1/AT-rich interaction domain 3 [ARID3] and Y47D3A.21/density-regulated re-initiation and release factor [DENR]) in C. elegans tissues using different auxins and transport inhibitor response 1 (TIR1)-expressing strains. We describe steps for genetic crossing, sample preparation, fluorescent microscopy, and treatment with either natural (indole-3-acetic acid [IAA]) or synthetic (1-naphthaleneacetic acid, potassium salt [K-NAA]) auxins. We then detail procedures for comparing the degree of CFI-1 depletion in C. elegans neurons upon panneuronal or pansomatic TIR1 expression. For complete details on the use and execution of this protocol, please refer to Li et al.1,2.
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Chronic graft-versus-host-disease (cGVHD) is one of the primary causes of morbidity and mortality for patients who undergo allogeneic hematopoietic stem cell transplantation (allo-HCT). In recent years, advancements in allo-HCT have allowed a broader range of patients to receive transplant, particularly older patients. We sought to assess the impact of cGVHD on outcomes in patients undergoing allo-HCT, for older patients as compared to their counterparts. We performed a retrospective analysis of all patients who underwent allo-HCT 1999-2018. Our results showed that those patients who developed cGVHD by D + 180 had an increased risk and incidence of NRM as compared to those patients without cGVHD. There was no significant difference in outcomes for those patients with cGVHD by age (≥60 years old [yo] and <60 yo). These findings suggest the significant morbidity of cGVHD, regardless of age.
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Obesity has erupted as an epidemic around the world. It has set itself as a fast wave among other prevailing specific clusters of non-communicable diseases. The current study reviews and presents an updated meaningful review of the vast research work performed at schools located in different cities of India. A systematic search was conducted in PubMed, Scopus, Google Scholar and PEDro. Studies representing data on obesity and overweight among children in Indian cities were included in the review. A total of 21 articles with 71,466 participants were included in the review for analysis. Obesity developed in childhood and adolescence is greatly associated with heart disease, stroke and cancer (breast and ovarian in women and prostate in men) in the late stage of life. In India, despite being a country with a faster rate of population becoming overweight and obese in urban areas, in contrast, rural areas are still struggling with malnutrition.
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Xenobiotic biotransformation is an important modulator of anthelmintic drug potency and a potential mechanism of anthelmintic resistance. Both the free-living nematode Caenorhabditis elegans and the ruminant parasite Haemonchus contortus biotransform benzimidazole drugs by glucose conjugation, likely catalysed by UDP-glycosyltransferase (UGT) enzymes. To identify C. elegans genes involved in benzimidazole drug detoxification, we first used a comparative phylogenetic analysis of UGTs from humans, C. elegans and H. contortus, combined with available RNAseq datasets to identify which of the 63 C. elegans ugt genes are most likely to be involved in benzimidazole drug biotransformation. RNA interference knockdown of 15 prioritized C. elegans genes identified those that sensitized animals to the benzimidazole derivative albendazole (ABZ). Genetic mutations subsequently revealed that loss of ugt-9 and ugt-11 had the strongest effects. The "ugt-9 cluster" includes these genes, together with six other closely related ugts. A CRISPR-Cas-9 deletion that removed seven of the eight ugt-9 cluster genes had greater ABZ sensitivity than the single largest-effect mutation. Furthermore, a double mutant of ugt-22 (which is not a member of the ugt-9 cluster) with the ugt-9 cluster deletion further increased ABZ sensitivity. This additivity of mutant phenotypes suggest that ugt genes act in parallel, which could have several, not mutually exclusive, explanations. ugt mutations have different effects with different benzimidazole derivatives, suggesting that enzymes with different specificities could together more efficiently detoxify drugs. Expression patterns of ugt-9, ugt-11 and ugt-22 gfp reporters differ and so likely act in different tissues which may, at least in part, explain their additive effects on drug potency. Overexpression of ugt-9 alone was sufficient to confer partial ABZ resistance, indicating increasing total UGT activity protects animals. In summary, our results suggest that the multiple UGT enzymes have overlapping but not completely redundant functions in benzimidazole drug detoxification and may represent "druggable" targets to improve benzimidazole drug potency.
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An easy, in situ growth approach led to the formation of several composites of metal-organic framewoks and Nb4C3Tx MXenes mixed intimately at the submicron scale. The high affinity of MXene surface for dopamine, enhanced by a nanostructuration induced by MOFs, resulted in superior sensing performances. The system exhibited good linearity over the 1-100 nM range, with an excellent limit of detection of 0.2 nM.
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Dopamina , Estruturas Metalorgânicas , Dopamina/análise , Dopamina/química , Estruturas Metalorgânicas/química , Limite de Detecção , Nióbio/químicaRESUMO
South Asians are a rapidly growing subset of the Asian population in the United States. They comprise people from multiple countries with diverse beliefs, languages, and cultural identities and values. The incidence of breast cancer is rising in South Asian women in the United States, with earlier onset and predilection for HER2-enriched tumors. Despite the rising incidence of breast cancer, participation in screening remains lower than other populations. Health care inequities in South Asian women are multifactorial and may be due to traditional health beliefs and practices, language barriers, cultural differences, and lack of overall awareness. Developing a culturally sensitive environment in breast imaging clinic practice can lead to improved patient care and adherence. Given the scarcity of data specific to the South Asian population in United States, there is a need for health service researchers and practice leaders to obtain more high-quality data to understand the needs of South Asian patient populations.
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BACKGROUND: Breast cancer is the most commonly diagnosed cancer among women globally, with significant impacts on physical, emotional, and functional well-being. Traditional rehabilitation methods may not fully address the multifaceted challenges faced by breast cancer survivors (BCSs), prompting exploration into innovative approaches such as Virtual Reality (VR) technology. OBJECTIVE: The present review aims to assess the effectiveness of VR in alleviating pain, improving Range of Motion (ROM), enhancing muscle strength, and augmenting the overall quality of life in patients undergoing breast cancer rehabilitation. METHODS: A comprehensive review of existing literature was conducted, focusing on studies investigating the use of VR in breast cancer rehabilitation. PubMed, Scopus, PEDro and Google scholar were searched for articles addressing VR interventions targeting pain management, ROM improvement, muscle strength enhancement, and quality of life enhancement in breast cancer patients. RESULTS: Findings yielded total 12 articles matching the selection criteria. VR technology has shown promising results in addressing the multifaceted needs of breast cancer patients. VR also serves as a distraction tool, positively impacting psychological well-being and mitigating negative psychological symptoms associated with the disease. CONCLUSION: VR represents a non-pharmacological approach to pain management and rehabilitation in breast cancer patients. Its ability to engage emotional, cognitive, and attention processes contributes to its effectiveness in enhancing overall quality of life. Further research is warranted to elucidate the long-term benefits and optimal utilization of VR technology in breast cancer rehabilitation programs.
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Plant breeding, aimed at enhancing desired traits, depends on genetic diversity. Mutation breeding is a powerful method of rapidly expanding genetic diversity, facilitating crop improvement, and ensuring food security. In a recent study, researchers evaluated the genetic variability of Trigonella species using different doses of sodium azide (SA) (0.2%, 0.4%, 0.6%, 0.8%, and 1.0%) through morphological, physiological, and cytogenetic studies. Morphological variations were observed in cotyledonary leaves, vegetative leaves, and overall plant growth and habit. Several quantitative parameters, such as plant height, fertile branches per plant, pods per plant (or clusters), seeds per pod, and seed yield, increased when treated with 0.2% and 0.4% SA compared to the control. Furthermore, the total chlorophyll content and carotenoids increased in the sample treated with 0.2% SA over the control but decreased with higher concentrations. Scanning electron microscopy revealed that stomatal aperture and seed dimensions increased at lower concentrations of sodium azide treatment. The study found a positive correlation between the different parameters studied in the Trigonella species, as indicated by high r-values. Based on their findings, it was concluded that the genotype of fenugreek can be improved by using 0.2% and 0.4% concentrations of sodium azide. However, the evaluation of observed variants in successive generations is a critical and necessary process to validate their potential as keystones for crop genetic improvements.
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The fear of an increase in blood sugar can be very traumatic. Being diabetic either type I or type II leads to a disorder called diabetes distress having traits of stress, depression, and anxiety. Among risk factors of diabetes mellitus heavy and trace metal toxicity emerges as new risk factors reported in many studies. In this study we target toxic metals, viz., Ni2+, Zn2+, and Cu2+, involved in the pathogenesis of diabetes and diabetic stress with naphthazarin esters. The compounds C1-C3 isolated from the leaves and roots of Arnebia guttata were tested for their metal-binding ability in an aqueous medium in UV-Visible and nuclear magnetic resonance (NMR) studies. These probes are well-known naphthoquinones present in the Arnebia species. In the UV-Visible titrations of compounds C1-C3 with Na2+, K2+, Zn2+, Ca2+, Cu2+, Mg2+, Co2+, and Ni2+ ions, significant binding was observed with Ni2+, Cu2+, and Zn2+ ions in MeOH/H2O. There occurs a beautiful formation of red-shifted bands between the 520 to 620 nm range with a synergistic increase in absorbance. Also, the disappearance of proton peaks in the 1H NMR spectrum on addition of metal ions confirmed binding. Compounds C1-C3 isolated from A. guttata came out as potent Ni2+, Zn2+, and Cu2+ sensors that are reportedly involved in islet function and induction of diabetes.
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Ésteres , Naftoquinonas , Ésteres/química , Naftoquinonas/química , Diabetes Mellitus/metabolismo , Neurotoxinas/química , Neurotoxinas/metabolismo , Água/química , Estrutura Molecular , Folhas de Planta/químicaRESUMO
Evolutionary psychology is the study of human psychological behavior. During childhood, men and women behave similarly; however, as a child approaches puberty, new physical and behavioral changes emerge. Behavioral psychology focuses on understanding the functioning and thought processes of the human mind. The general population lacks knowledge of basic behavioral differences between men and women, leaving them unaware of their role, limitations, societal responsibilities, resulting in an underestimation of their own natural talents and biology. Thus, people tend to follow societal norms rather than exploring and utilizing their natural talents. The current review was designed and conducted to enforce compression on behavioral psychology in both genders as well as to identify variations in hormonal activity and sexual preferences.
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Classically, Qigong is a Chinese technique that has been practiced in China for the past 3,000 years for healing the inner self. Qigong, wherein "Qi" means body energy and "Gong" denotes cultivation, regulates the energy flow in the body. The Qigong technique comprises a package of deep breathing training, gentle and rhythmic movement, and muscle-strengthening exercises that heal the body to activate one's internal soul energy. It has demonstrated its efficacy by inducing relaxation, building up stamina, strengthening immunity, appreciating muscle conditioning, and minimizing anxiety and depression. Furthermore, it has been beneficial in improving awareness of joint and movement senses. Specifically, Qigong brings healing by regulating energy flow in the whole-body systems. Moreover, it has exhibited a variety of regenerating effects by inducing emotional and mental relaxation. In today's world, Qigong exercises are being used for treating musculoskeletal disorders that are work- and stress-related by nature. Qigong is practiced globally as deep breathing exercises, and meditation is practiced for peace of mind and spirituality, whereas vigorous practice includes martial arts.
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The role of diet in the development of skin disorders is well-established, with nutritional deficiency often identified as a risk factor for skin diseases. Imbalances in the skin can be caused by nutritional deficiencies, excessive intake, insufficient nutrients, and hazardous ingredients. Patients frequently inquire about the impact of dietary patterns on skin health when consulting dermatologists in clinical settings. Simultaneously, the popularity of using nutritional supplements containing vitamins, minerals, and nutraceutical blends has been on the rise. It is crucial for dermatologists, primary care physicians, and other healthcare providers to be acquainted with evidence-based dietary interventions, distinguishing them from those that are more market-driven than truly efficacious. This review explores the modification of diet, encompassing both dietary exclusion and supplementation, as a therapeutic approach for conditions such as psoriasis, atopic dermatitis, bullous disease, vitiligo, and alopecia areata. A comprehensive literature search, utilizing the PubMed/Medline, Google Scholar, and Medscape databases, was conducted to investigate the relationship between each nutrient and various inflammatory skin diseases. The findings emphasize the significance of a well-balanced and thoughtfully planned diet in supplying adequate amounts of proteins, vitamins, and minerals to support optimal skin health. Additionally, this comprehensive review navigates through various dietary recommendations, offering insights into their multifaceted impacts on the immune system, gut microbiome, and skin health. The goal is to pave the way for informed and targeted dietary interventions for individuals dealing with food allergies and associated skin conditions.
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BACKGROUND: Mesenchymal stem/stromal cells (MSCs) can regenerate tissues through engraftment and differentiation but also via paracrine signalling via extracellular vesicles (EVs). Fetal-derived MSCs (fMSCs) have been shown, both in vitro and in animal studies, to be more efficient than adult MSC (aMSCs) in generating bone and muscle but the underlying reason for this difference has not yet been clearly elucidated. In this study, we aimed to systematically investigate the differences between fetal and adult MSCs and MSC-derived EVs at the phenotypic, RNA, and protein levels. METHODS: We carried out a detailed and comparative characterization of culture-expanded fetal liver derived MSCs (fMSCs) and adult bone marrow derived MSCs (aMSCs) phenotypically, and the MSCs and MSC-derived EVs were analysed using transcriptomics and proteomics approaches with RNA Sequencing and Mass Spectrometry. RESULTS: Fetal MSCs were smaller, exhibited increased proliferation and colony-forming capacity, delayed onset of senescence, and demonstrated superior osteoblast differentiation capability compared to their adult counterparts. Gene Ontology analysis revealed that fMSCs displayed upregulated gene sets such as "Positive regulation of stem cell populations", "Maintenance of stemness" and "Muscle cell development/contraction/Myogenesis" in comparison to aMSCs. Conversely, aMSCs displayed upregulated gene sets such as "Complement cascade", "Adipogenesis", "Extracellular matrix glycoproteins" and "Cellular metabolism", and on the protein level, "Epithelial cell differentiation" pathways. Signalling entropy analysis suggested that fMSCs exhibit higher signalling promiscuity and hence, higher potency than aMSCs. Gene ontology comparisons revealed that fetal MSC-derived EVs (fEVs) were enriched for "Collagen fibril organization", "Protein folding", and "Response to transforming growth factor beta" compared to adult MSC-derived EVs (aEVs), whereas no significant difference in protein expression in aEVs compared to fEVs could be detected. CONCLUSIONS: This study provides detailed and systematic insight into the differences between fMSCs and aMSCs, and MSC-derived EVs. The key finding across phenotypic, transcriptomic and proteomic levels is that fMSCs exhibit higher potency than aMSCs, meaning they are in a more undifferentiated state. Additionally, fMSCs and fMSC-derived EVs may possess greater bone forming capacity compared to aMSCs. Therefore, using fMSCs may lead to better treatment efficacy, especially in musculoskeletal diseases.
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Vesículas Extracelulares , Células-Tronco Mesenquimais , Animais , Transcriptoma , Proteômica , Células-Tronco Mesenquimais/metabolismo , Perfilação da Expressão Gênica , Vesículas Extracelulares/metabolismoRESUMO
Objectives: The study aimed to compare the effect of cranial electrical stimulation (CES) and transcranial direct current stimulation (tDCS) in improving cognition among individuals with mild traumatic brain injury. Patients and methods: The pretest-posttest randomized controlled study was conducted between November 2020 and March 2022. Seventy-two patients (64 males, 8 females; mean age: 40.5±9.5 years; range, 18 to 45 years) experiencing cognitive impairment within three months of traumatic brain injury were recruited. Participants were randomly assigned into two groups: Group 1 (CES with cognitive training, n=36) and Group 2 (tDCS with cognitive training, n=36). Participants were blinded in the study. Both groups received 30-min sessions of neuromodulation along with 30 min of cognitive training five days a week for four weeks. The patients were assessed at baseline and at the end of two and four weeks of intervention. The primary outcome measure was the Montreal Cognition Assessment (MoCA), and the secondary outcome measure was the Galveston Orientation Amnesia Test (GOAT). Results: Demographic and baseline characteristics depicted normal distribution for both groups (p>0.05). Within group analyses of both groups demonstrated significant differences for both outcome measures (MoCA: p=0.001; GOAT: p=0.001). Between group analyses of MoCA showed significant improvement with p-value of 0.001 while GOAT exhibited p-value of 0.002 showing significant difference between the two groups. Time group interaction effect and covariance analyses depicted significant improvement with p-value of 0.001 for both outcome measures with excellent effect size >0.80. Conclusion: Cranial electrical stimulation was a more effective noninvasive neuromodulatory device than tDCS in improving cognition among individuals with traumatic brain injury.
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BACKGROUND: Chemoradiation in head and neck carcinoma (HNC) shows significant anatomical resulting in erroneous dose deposition in the target or the organ at risk (OAR). Adaptive radiotherapy (ART) can overcome this. Timing of significant target and OAR changes with dosimetric impact; thus, most suitable time and frequency of ART is unclear. METHODS: This dosimetric study used prospective weekly non-contrast CT scans in 12 HNC patients (78 scans). OARs and TVs were manually contoured after registration with simulation scan. Dose overlay done on each scan without reoptimization. Dosimetric and volumetric variations assessed. RESULTS: Commonest site was oropharynx. Gross Tumor Volume (GTV) reduced from 47.5 ± 19.2 to 17.8 ± 10.7 cc. Nodal GTV reduced from 15.7 ± 18.8 to 4.7 ± 7.1 cc. Parotid showed mean volume loss of 35%. T stage moderately correlated with GTV regression. CONCLUSION: Maximum GTV changes occurred after 3 weeks. Best time to do single fixed interval ART would be by the end of 3 weeks.