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Phakomatoses, or neurocutaneous syndromes, are a heterogeneous group of rare genetic disorders that predominantly affect structures arising from the embryonic ectoderm, namely the skin, eye globe, retina, tooth enamel, and central nervous system. Other organs are also involved in some syndromes, mainly cardiovascular, pulmonary, renal, and musculoskeletal systems. Currently, more than sixty distinct entities belonging to this category have been described in the literature. Common phakomatoses include conditions like Neurofibromatosis and Tuberous sclerosis. Several review papers have focused on various aspects of these common conditions, including clinical presentation, genetic and molecular basis, and neuroimaging features. In this review, we focus on rare neurocutaneous syndromes: Melanophakomatoses (i.e., Neurocutaneous Melanosis, and Incontinentia Pigmenti), Vascular Phakomatoses (i.e., Ataxia Telangiectasia and PHACE Syndrome), and other conditions such as Cowden Syndrome, Basal Nevus Syndrome, Schwannomatosis, Progressive Facial Hemiatrophy, Gomez-Lopez-Hernandez Syndrome, Wyburn-Mason Syndrome, CHILD Syndrome, and Proteus Syndrome. We also review the neuroradiologic manifestations of these conditions as a guide for neurologists and neuroradiologists in their daily practice.

Temporal lobe epilepsy (TLE) and depression are common comorbid disorders whose underlying shared neural network has yet to be determined. Although animal studies demonstrate a role for the dorsal bed nucleus of the stria terminalis (dBNST) in both seizures and depression, and human clinical studies demonstrate a therapeutic effect of stimulating this region on treatment-resistant depression, the role of the dBNST in depressed and nondepressed TLE patients is still unclear. Here, we tested the hypothesis that this structure is morphologically abnormal in these epilepsy patients, with an increased abnormality in TLE patients with comorbid depression.In this case-controlled study, 3-T structural magnetic resonance imaging scans were obtained from TLE patients with no depression (TLEonly), TLE patients with depression (TLEdep), and healthy control (HC) subjects. TLE subjects were recruited from the Yale University Comprehensive Epilepsy Center, diagnosed with the International League Against Epilepsy 2014 Diagnostic Guidelines, and confirmed by video-electroencephalography. Diagnosis of major depressive disorder was confirmed by a trained neuropsychologist through a Mini International Neuropsychiatric Interview based on the Diagnostic and Statistical Manual of Mental Disorders, 4th edition. The dBNST was delineated manually by reliable raters using Bioimage Suite software.The number of patients and subjects included 35 TLEonly patients, 20 TLEdep patients, and 102 HC subjects. Both TLEonly and TLEdep patients had higher dBNST volumes compared to HC subjects, unilaterally in the left hemisphere in the TLEonly patients (p = .003) and bilaterally in the TLEdep patients (p .0001). Furthermore, the TLEdep patients had a higher dBNST volume than the TLEonly patients in the right hemisphere (p = .02).Here, we demonstrate an abnormality of the dBNST in TLE patients, both without depression (left enlargement) and with depression (bilateral enlargement). Our results demonstrate this region to underlie TLE both with and without depression, implicating it as a target in treating the comorbidity between these two disorders.

Tyrosine kinase inhibitors (TKIs) such as imatinib, nilotinib, dasatinib, and ponatinib have significantly improved the life expectancy of Philadelphia chromosome-positive (Ph+) acute lymphocytic leukemia (ALL) patients; however, resistance to TKIs remains a major clinical challenge. Point mutations in the tyrosine kinase domain (TKD) of BCR-ABL1 have emerged as the predominant cause of acquired resistance. In approximately 30% of patients, the mechanism of resistance to TKIs remains elusive. This study aimed to investigate mechanisms of nonmutational resistance in Ph+ ALL. Here we report the development of a nonmutational resistance cell line SupB15-RT; conferring resistance to approved ABL kinase inhibitors (AKIs) and allosteric inhibitors GNF-2, ABL001, and crizotinib, except for dasatinib (IC90 50nM), a multitarget kinase inhibitor. We found that the AKT/mTOR pathway is activated in these cells and their proliferation inhibited by Torin-1 with an IC50 of 24.7 nM. These observations were confirmed using 3 different ALL patient-derived long term cultures (PDLTCs): (1) HP (BCR-ABL1 negative), (2) PH (BCR-ABL1 positive and responsive to TKIs) and (3) BV (BCR-ABL1 positive and nonmutational resistant to TKIs). Furthermore, Torin-1 and NVP-BEZ235 induced apoptosis in PH and BV cells but not in HP cells. Our experiments provide evidence of the involvement of AKT/mTOR pathway in the evolution of nonmutational resistance in Ph+ ALL which will assist in developing novel targeted therapy for Ph+ ALL patients with BCR-ABL1 independent nonmutational resistance.

Background: In humans, the β-like globin genes are encoded from a single locus comprising five globin genes (ε-, Gγ-, Aγ-, δ-, and β-globin in sequence) and their expression is under developmental control. The γ-globin genes (Gγ- and Aγ-) are expressed during fetal life and replaced by adult β-globin after birth. Mutations in the β-globin gene cause β-hemoglobinopathies such as sickle cell disease (SCD) and β-thalassemia. The clinical severity of SCD and β-thalassemia can be mitigated by elevated fetal hemoglobin (HbF) levels, which have been found in individuals with the benign hereditary persistence of fetal hemoglobin (HPFH) syndrome. Thus, reactivating the expression of γ-globin genes is an attractive treatment strategy for β-hemoglobinopathies. Reactivation of γ-globin expression by disrupting the binding of the BCL11A transcriptional repressor complex to the γ-globin gene promoter, provides a novel approach for inducing fetal hemoglobin. Methodology: The BCL11A binds a 5′-TGACCA-3′ element (spanning nucleotides −118 to −113) of globin genes using its three C-terminal Zinc fingers (Znf). The binding efficiency of Znf4, Znf5, and Znf6 was predicted by using in silico tools (SIFT, SNAP, PolyPhen-2, PANTHER, I-Mutant, PROVEAN, SNPs&GO, mCSM, and PhD-SNP), molecular dynamic simulation and homology modelling. K562 cells were electroporated with CRISPR-Cas9 targeting the BCL11A Znf4. Results: The binding energy scores illustrate that deleting Znf5 and Znf6 decreased the binding affinity by ~3.7-fold, whereas deleting Znf4 decreased the affinity by > 70-fold. Using CRISPR-Cas9 genome-editing strategy, we deleted Znf4 (260bp genomic region) within the BCL11A in K562 cell lines. Znf4 deletion resulted in a readily detectable γ-globin increase with a preferential increase in G-gamma. Conclusion: Altogether, our findings highlight the valuable insights for improving gene editing therapy strategies by either deleting Znf4 of BCL11A or the TTGACCA motif to disrupt the interaction between BCL11A and other interacting partners and the γ-globin gene promoter. Complete failure in the protein-protein interactions with functional partners and to the γ-globin gene promoter revealed that Znf4 is a suitable target for disrupting BCL11A-mediated hemoglobin switching. Citation Format: Irfan Hussain, Kainaat Mumtaz, Hammad Hassan, Afsar Ali Mian. Amelioration of hemoglobinopathies by targeted deletion of zinc finger domain within BCL11A gene using CRISPR-Cas9 technology. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4746.

Background: Exosomes are extracellular vesicles having less than 200 nm size, secreted extracellularly by almost all types of cells. They regulate intercellular communication by delivering RNAi, proteins, viruses, and soluble factors as inherited payloads. In recent years, the application of exosomes to deliver small molecules have been widely investigated in tissue regeneration, therapeutics, as well as diagnostics. However, the isolation of exosomes has remained challenging mainly due to limitations of specialized instruments and the least cost-effective reagents. In this study, we developed a novel method to isolate exosomes from cultured cells. Experimental: The supernatant from cultured cells was collected and subjected to lyophilization after low-speed centrifugation. The characterization and quantification of exosomes were performed using Bradford assay. Scanning Electron Microscopy (SEM) and Nanoparticles Tracking Analysis (NTA) were carried out for shape and size distribution. Western blot was performed for the evaluation of the exosome specific surface markers (CD63, CD9, CD81, calnexin, HSP70, and TSG101). The as-isolated exosomes were loaded with commercial dyes (Cy5, Eosin) for the evaluation of their drug delivery properties. Furthermore, their drug loading and release were confirmed by loading tyrosine kinase inhibitors (TKIs) as a payload. Finally, the anti-leukemic drugs (dasatinib and ponatinib) were loaded on the isolated exosomes for targeting leukemia cell line i.e., K562 and BCR-ABL expressing Ba/F3 cells in in vitro model, using MTT assay for cell proliferation and Annexin-V/PI for apoptosis detection. Results: Here we found that the isolated exosomes have round shape with an average size of ~130 nm. They expressed known exosome markers including CD63, CD9, CD81, calnexin, HSP70, and TSG101. These exosomes could efficiently target and deliver dyes (Eosin and Cy5) and TKIs (dasatinib and ponatinib) to K562 and Ba/F3-cells expressing BCR-ABL. The TKIs-loaded exosomes inhibited the growth of leukemic cells and induced apoptosis in a dose dependent manner. Conclusion: We claim the development of a new method for active exosome isolation from cell culture. These exosomes could qualify for all the essential characterization, drug loading, and release ability by harnessing proliferation and inducing apoptosis in Ph+ leukemia cells. Citation Format: Fawad Ur Rehman, Rida e Maria Qazi, Zahra Sajid, Afsar Ali Mian. Exosomes isolation via lyophilization for Philadelphia-positive leukemia therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 833.

Background: Asciminib an allosteric tyrosine kinase inhibitor (TKI) was recently approved by the FDA for managing Philadelphia positive chronic myeloid leukemia (CML) patients in chronic phase who had failed to respond to previous TKIs. In this study, we used human erythroleukemic K562 cell line to investigate the role of asciminib as an inducer of erythroid differentiation. Additionally, imatinib, which has previously been reported as a potential inducer of erythroid differentiation, was also tested on these cells, both independently as well as in combination, in order to investigate the degree of erythroid differentiation in a K562 cell line. Methodology: In this study, we investigated the effect of asciminib (5-20nM) with and without imatinib (50-200nM) in differentiating the erythroleukemic cell line K562 towards erythroid lineage. K562 cells were cultured over a 12-day period in basic (IMDM) media and an EPO-based differentiation media to select optimal medium conditions. In addition, an in-vitro drug cytotoxicity experiment (XTT) was used to determine the preliminary toxicity of asciminib with IMDM media and EPO-based differentiation media CD235a and CD71 surface marker expressions were analyzed by flow cytometry at all time-points to determine the degree of differentiation achieved by each of the conditions Moreover, to analyze and identify hemoglobin producing cells ensuing differentiation, benzidine staining was performed at indicated time points. Results: Our results suggest that asciminib, imatinib or a combination of both drugs in EPO-based differentiation media results in efficient erythroid differentiation with high levels of GPA expression (approx. 90%) and significant upregulation of globin genes (p value = 0.0001). These results were further confirmed by benzidine staining, where drug addition and dose escalation significantly increased benzidine-positive cells. Our results also demonstrated that asciminib was able to achieve maximum differentiation at a much earlier time point as compared to imatinib. Moreover, asciminib, when combined with another TKI such as imatinib, can also aid in lowering the drug concentrations being used, hence resulting in lessening the toxicity of the drug when used alone required at high doses. Conclusion: The results from this study highlighted the novel role of asciminib in erythroid differentiation using an in-vitro K562 model that can give high-levels of surface markers and globin expression. Potential clinical application of asciminib in hematological diseases that are associated specifically with a failure in the expression of globin genes should be evaluated. Keywords:K562, erythroid differentiation, asciminib, imatinib, glycophorin A, transferrin receptor, hemoglobin Citation Format: Hammad Hassan, Sarah Yousuf, Syed Muhammad Ahmed, Karim Ruknuddin, Hadiqa Raees, Sadia Habib, Fizza Iftikhar, Afsar Ali Mian, Elnasir Lalani. Asciminib activates erythroid differentiation in K562 cell line. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4040.

Coronaviruses are a family of viruses that can be transmitted from one person to another Earlier strains have only been mild viruses, but the current form, known as coronavirus disease 2019 (COVID-19), has become a deadly infection The outbreak originated in Wuhan, China, and has since spread worldwide The symptoms of COVID-19 include a dry cough, sore throat, fever, and nasal congestion Antimicrobial drugs, pathogen-host interaction, and 2 weeks of isolation have been recommended for the treatment of the infection Safe operating procedures, such as the use of face masks, hand sanitizer, handwashing with soap, and social distancing, are also suggested Moreover, travel bans for cities, states, and countries have been put in place, along with lockdowns to control the outbreak Travel restrictions, mask use, sanitizer or soap use, and avoidance of touching the face and nose have produced encouraging results, whereas the effectiveness of antibiotics has not been proved The results of isolation for the recovery of infected people have also been promising Travel bans and lockdowns have caused a slump in economies, and unemployment has risen sharply, resulting in an increase in mental health cases globally To date, vaccines have been developed and are in use in certain countries, but following standard operating procedures remain critical The countries following the guidelines can eradicate this virus New Zealand was the first country to eliminate the virus from their territory

COVID-19 is a pandemic faced by almost every country in the world, this has resulted in health crisis. Due to COVID-19, all the countries around the world have decided to close all educational institutes to prevent this pandemic. Educational institutes have taken every possible measure to minimize the impact of the closure of schools and introduce the concept of an online education system which is not only a massive shock for parents but it also affects the children's learning process and social life. The educational applications (Apps) are very important, because they offer more opportunities for development and growth to society. In this pandemic situation, educational Apps like Zoom, HEC LMS, Google Classroom, and Skype, etc. are the need of the hour when everything goes online. In this paper, the usability features of online educational Apps are thoroughly discussed including the effectiveness and usability for students. Using the results obtained from the survey, this paper observes the student's perspective of usefulness of online educational Apps in student’s learning process of different age groups. It also analyzes the easiness for students to understand, interact and use these Apps.

The Internet-of-Things (IoT) has become an important topic among researchers owing to its potential to change the way we live and use smart devices. In recent years, many research work found in the world are interrelated and convey via the existing web structure which makes a worldwide system called IoT. This study focused on the significant improvement of answers for a wider scope of gadgets and the Internet of Things IoT stages in recent years. In any case, each arrangement gives its very own IoT framework, gadgets, APIs, and information configurations promoting interoperability issues. These issues are the outcome of numerous basic issues, difficulty to create IoT application uncovering cross-stage, and additionally cross-space, trouble in connecting non-interoperable IoT gadgets to various IoT stages, what's more, eventually averts the development of IoT innovation at an enormous scale. To authorize consistent data sharing between various IoT vendors, endeavors by a few academia, industrial, and institutional groups have accelerated to support IoT interoperability. This paper plays out a far-reaching study on the cutting-edge answers for encouraging interoperability between various IoT stages. Likewise, the key difficulties in this theme are introduced.