Crosstalk among WEE1 Kinase, AKT, and GSK3 in Nav1.2 Channelosome Regulation.

The signaling complex around voltage-gated sodium (Nav) channels includes accessory proteins and kinases crucial for regulating neuronal firing. Previous studies showed that one such kinase, WEE1-critical to the cell cycle-selectively modulates Nav1.2 channel activity through the accessory protein fibroblast growth factor 14 (FGF14). Here, we tested whether WEE1 exhibits crosstalk with the AKT/GSK3 kinase pathway for coordinated regulation of FGF14/Nav1.2 channel complex assembly and function. Using the in-cell split luciferase complementation assay (LCA), we found that the WEE1 inhibitor II and GSK3 inhibitor XIII reduce the FGF14/Nav1.2 complex formation, while the AKT inhibitor triciribine increases it. However, combining WEE1 inhibitor II with either one of the other two inhibitors abolished its effect on the FGF14/Nav1.2 complex formation. Whole-cell voltage-clamp recordings of sodium currents (INa) in HEK293 cells co-expressing Nav1.2 channels and FGF14-GFP showed that WEE1 inhibitor II significantly suppresses peak INa density, both alone and in the presence of triciribine or GSK3 inhibitor XIII, despite the latter inhibitor's opposite effects on INa. Additionally, WEE1 inhibitor II slowed the tau of fast inactivation and caused depolarizing shifts in the voltage dependence of activation and inactivation. These phenotypes either prevailed or were additive when combined with triciribine but were outcompeted when both WEE1 inhibitor II and GSK3 inhibitor XIII were present. Concerted regulation by WEE1 inhibitor II, triciribine, and GSK3 inhibitor XIII was also observed in long-term inactivation and use dependency of Nav1.2 currents. Overall, these findings suggest a complex role for WEE1 kinase-in concert with the AKT/GSK3 pathway-in regulating the Nav1.2 channelosome.

Funding Pattern Comparison Pre- and Post-Ayushman Bharat National Health Scheme (AB-PMJAY) in Active Females Undergoing TKA for Advanced Knee Arthritis with Clinical, QoL Analysis and AB-PMJAY Technical Data.

Introduction: Patients visiting for Knee Arthroplasty have often been treated at neighborhood clinics and bonesetters. India floated world's largest publicly funded national health insurance program -AB-PMJAY covering Knee Arthroplasty. AB-PMJAY's data for Arthroplasty has not been published. Methods: A Prospective study from Jan 2016- Jan 2023 on females undergoing TKA. Age, DEXA Score, Walking Ability, KSS, 10MWT, SF36 Scores, Funding Pattern before and after AB-PMJAY, Time to Approval, Time to discharge, Time to Query reply and Rejection Rates were documented. Results: 790 patients (91.86%) received treatment previously. 650 (78.54%) patients lived with family & 32 patients lost to follow-up. 37.67% & 62.32% patients had Grade. 3 & 4 osteoarthritis respectively. Commonest comorbidity was Vitamin D deficiency followed by Menopause and Hypertension. Post-treatment Vitamin D deficiency reduced from 68.59% to 2.17% at 3 months. Post-surgery, Functional Scores improved significantly at 1st and 6th month. VAS dropped significantly from 7.8 to 3.6 at 1st month follow up. At 6 months, 81.52% patients were independent outdoor walkers compared to 9.42% (baseline). Average hospital stay-54 hours and Return to work- 42 days. Central health scheme beneficiaries rose significantly from 12% (pre-PMJAY) to 42% (post-PMJAY). With AB-PMJAY, no rejection for surgical procedures, Time to surgery approval was 16 hours (mean) and Time to Discharge was 8 hours (mean). Time to reply was 6 hours (mean). Conclusion: Evident rise in patients getting TKA done through AB-PMJAY. TKA has become an affordable and achievable target for financially deprived patients, post regulation of knee implant prices & AB-PMJAY.

Mechanistic Investigation of Thiazole-Based Pyruvate Kinase M2 Inhibitor Causing Tumor Regression in Triple-Negative Breast Cancer.

Triple-negative breast cancer (TNBC) is a deadly breast cancer with a poor prognosis. Pyruvate kinase M2 (PKM2), a key rate-limiting enzyme in glycolysis, is abnormally highly expressed in TNBC. Overexpressed PKM2 amplifies glucose uptake, enhances lactate production, and suppresses autophagy, thereby expediting the progression of oncogenic processes. A high mortality rate demands novel chemotherapeutic regimens at once. Herein, we report the rational development of an imidazopyridine-based thiazole derivative 7d as an anticancer agent inhibiting PKM2. Nanomolar range PKM2 inhibitors with favorable drug-like properties emerged through enzyme assays. Experiments on two-dimensional (2D)/three-dimensional (3D) cell cultures, lactate release assay, surface plasmon resonance (SPR), and quantitative real-time polymerase chain reaction (qRT-PCR) validated 7d preclinically. In vivo, 7d outperformed lapatinib in tumor regression. This investigation introduces a lead-based approach characterized by its clear-cut chemistry and robust efficacy in designing an exceptionally potent inhibitor targeting PKM2, with a focus on combating TNBC.

Encyclopaedic Review of Glipizide Pre-clinical and Clinical Status.

Glipizide is an oral glucose-lowering medication that is beneficial for the treatment of type 2 diabetes. This study compiles exhaustively all accessible information on glipizide, from preclinical to clinical studies. Glipizide may be used in concert with TRAIL to treat cancer cells; in vitro studies have shown that it suppresses angiogenesis and vasculogenesis while shielding cells from glycation-induced damage. Anticonvulsant effects and modifications in the pharmacokinetics of other medications, such as Divalproex Sodium, were seen in glipizide in vivo experiments. Propranolol amplifies glipizide's hypoglycemic effect briefly in normal animals but consistently enhances it in diabetic ones. In the treatment of cancer and neurodegenerative poly(Q) illnesses, glipizide has demonstrated to offer potential therapeutic advantages. It is ineffective in preventing DENA-induced liver cancer and may cause DNA damage over time. The way glipizide interacts with genetic variants may increase the risk of hypoglycemia. Combining Syzygium cumini and ARBE to glipizide may enhance glycemic and lipid control in type 2 diabetes. Individuals with coronary artery disease who take glipizide or glyburide have an increased risk of death. The risk of muscular responses and acute pancreatitis is minimal when glipizide and dulaglutide are combined. In conclusion, glipizide has shown promising therapeutic efficacy across a variety of disorders.

Dendrimers: Patents for Alzheimer's Disease.

Cells and nervous system connections that are crucial for movement, coordination, strength, sensation, and thought are gradually damaged in neurodegenerative illnesses. Amyloid beta (Aß)- accumulating macromolecules in the brain are the primary cause of the disease's chronic symptoms, according to analysis carried out during the last 20 years. Plaques and clumps of amyloid- build up in the brain, obstructing neuronal signals and destroying neural connections. Tau, a protein that results in the formation of "neurofibrillary tangles" in the brain, another hallmark of neuronal death, has been the focus of a lot of research. Dendrimers Delivery (DDs) is one of the most promising advancements in nanotechnology for biomedical applications, particularly drug delivery. Some of the main categories of dendrimers employed in the successful management of neurodegenerative illnesses are polyamidoamine dendrimers (PAMAM) dendrimers, polypropylenimine dendrimers (PPI), Poly-l-lysine dendrimers (PLL), and carbosilane dendrimers. The tight blood-brain barrier (BBB), which limits the entry of medications or therapeutic agents, makes it difficult to treat central nervous system disorders. Dendrimers have attracted the attention of scientists more than other non-invasive methods of drug delivery across the BBB and improve the uptake of medicines in the brain's target tissues. The major benefits of dendrimers include their adaptability, biocompatibility, ability to load pharmaceuticals into the core and surface, and nanosize. This review has updated the status of the patent and clinical trials literature pertaining to dendrimer use in AD.

Therapy-Acquired Clonal Mutations in Thiopurine Drug-Response Genes Drive Majority of Early Relapses in Pediatric B-Cell Precursor Acute Lymphoblastic Leukemia.

METHODS: Forty pediatric (0-12 years) B-ALL DNA samples (20 paired Diagnosis-Relapse) and an additional six B-ALL DNA samples (without relapse at 3 years post treatment), as the non-relapse arm, were retrieved from the biobank for advanced genomic analysis. Deep sequencing (1050-5000X; mean 1600X) was performed using a custom NGS panel of 74 genes incorporating unique molecular barcodes. RESULTS: A total 47 major clones (>25% VAF) and 188 minor clones were noted in 40 cases after bioinformatic data filtering. Of the forty-seven major clones, eight (17%) were diagnosis-specific, seventeen (36%) were relapse-specific and 11 (23%) were shared. In the control arm, no pathogenic major clone was noted in any of the six samples. The most common clonal evolution pattern observed was therapy-acquired (TA), with 9/20 (45%), followed by M-M, with 5/20 (25%), m-M, with 4/20 (20%) and unclassified (UNC) 2/20 (10%). The TA clonal pattern was predominant in early relapses 7/12 (58%), with 71% (5/7) having major clonal mutations in the NT5C2 or PMS2 gene related to thiopurine-dose response. In addition, 60% (3/5) of these cases were preceded by an initial hit in the epigenetic regulator, KMT2D. Mutations in common relapse-enriched genes comprised 33% of the very early relapses, 50% of the early and 40% of the late relapses. Overall, 14/46 (30%) of the samples showed the hypermutation phenotype, of which the majority (50%) had a TA pattern of relapse. CONCLUSIONS: Our study highlights the high frequency of early relapses driven by TA clones, demonstrating the need to identify their early rise during chemotherapy by digital PCR.

Association of COVID-19 with Comorbidities: An Update.

Coronavirus disease (COVID-19) is caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) which was identified in Wuhan, China in December 2019 and jeopardized human lives. It spreads at an unprecedented rate worldwide, with serious and still-unfolding health conditions and economic ramifications. Based on the clinical investigations, the severity of COVID-19 appears to be highly variable, ranging from mild to severe infections including the death of an infected individual. To add to this, patients with comorbid conditions such as age or concomitant illnesses are significant predictors of the disease's severity and progression. SARS-CoV-2 enters inside the host cells through ACE2 (angiotensin converting enzyme2) receptor expression; therefore, comorbidities associated with higher ACE2 expression may enhance the virus entry and the severity of COVID-19 infection. It has already been recognized that age-related comorbidities such as Parkinson's disease, cancer, diabetes, and cardiovascular diseases may lead to life-threatening illnesses in COVID-19-infected patients. COVID-19 infection results in the excessive release of cytokines, called "cytokine storm", which causes the worsening of comorbid disease conditions. Different mechanisms of COVID-19 infections leading to intensive care unit (ICU) admissions or deaths have been hypothesized. This review provides insights into the relationship between various comorbidities and COVID-19 infection. We further discuss the potential pathophysiological correlation between COVID-19 disease and comorbidities with the medical interventions for comorbid patients. Toward the end, different therapeutic options have been discussed for COVID-19-infected comorbid patients.

Cytology as the guiding light: Prompt cytomorphologic diagnosis of cystic metastases from a jejunal neuroendocrine tumor masquerading as hepatic abscesses.

Hepatic cystic lesions are commonly seen in radiology and can occur due to infective or neoplastic causes. It is imperative to determine the precise nature of these hepatic cysts owing to significant therapeutic and prognostic differences. Hepatic metastases from gastrointestinal neuroendocrine tumors (NETs) mostly present as solid hypervascular lesions. Cystic presentation of the metastatic NETs is extremely rare. A 64-year-old woman presented with an abdominal lump of 3 months duration and high-grade fever from the last 2 days. An ultrasound abdomen revealed multiple hypoechoic liver lesions suggestive of liver abscesses. An ultrasound-guided fine-needle aspiration (FNA) from the cystic liver lesions revealed a NET confirmed by immunocytochemistry. The Ki-67 index was 5%; hence, a final diagnosis of metastatic NET, grade 2, was given with advice to work up for localizing the primary tumor. Subsequently, a contrast-enhanced computerized tomography of the abdomen revealed a polypoidal lesion in the proximal jejunum, which showed intense expression on the whole-body DOTANOC scan (SUVmax-76.5), indicating it to be the likely primary tumor along with somatostatin receptor-expressing multiple lymph nodes and hypodense liver lesions. Thus, a final diagnosis of a jejunal NET with abdominal lymph node and liver metastases was made. The present report highlights the importance of FNA as a reliable modality in diagnosing metastatic NETs presenting as unusual cystic hepatic metastases masquerading as liver abscesses. Additionally, the cell block helps confirm the cytologic diagnosis and enables upfront tumor grading, thereby helping in prognostication and therapeutic decision-making.

Self-regulated cobalt zinc ferrite system as a potential nanoplatform for the synergistic effect of hyperthermia-chemo agent for cancer therapy.

Magnetic hyperthermia (MH) has been studied for almost seventy-five years, but its efficacy in clinical applications is still fiercely contested. Despite this, few magnetic nanosystems are approved for clinical usage due to their strong affinity as drug carriers. The most important condition for hyperthermia applications for successful cancer therapy is magnetic nanoparticles with a controlled heating pattern (42-46 °C) for a prolonged timeframe. In the current study, cobalt-zinc nanoferrites (MNPs) having a Curie temperature of 46 â„ƒ with a tunable heating profile was loaded with Doxorubicin (DOX) through a surface conjugation technique (DOX-Cs-MNPs), and characterized by using multiple techniques. The magnetic hyterises (M-H) curves revealed the occurrence of superparamagnetism in the MNPs with extremely low coercivity; further, the DOX-loaded nanoparticles exhibited enhanced saturation magnetization. More importantly, the MNPs showed that they could maintain a therapeutic temperature for an indefinite amount of time. High drug loading affinity (86 %) was observed on MNPs with pH and temperature-controlled release. Under in vitro conditions, the biocompatible DOX-Cs-MNPs caused substantial apoptosis in MCF-7 cells (72 %) with overall cell death of < 95 %. The distinctive MNPs thus have the potential to be used in clinical applications.

The Diagnostic Accuracy of Serum and Urine Human Epididymis Protein 4 (HE4) in Ovarian Cancer in 15,394 Subjects: An Updated Meta-Analysis.

Background We aim to determine the diagnostic accuracy of both serum and urinary human epididymis protein 4 (HE4) in the diagnosis of ovarian cancer. Methods Electronic databases and search engines such as PubMed, Cochrane Library, and Google Scholar were searched systematically by two independent reviewers to retrieve articles published from inception to June 11, 2022. The diagnostic accuracy of serum and urinary HE4 was computed using the random-effects model in terms of pooled sensitivity, pooled specificity, and diagnostic odds ratio (DOR) with 95% confidence interval (CI). To explain any source of possible heterogeneity, meta-regression and subgroup analyses were performed. Risk of bias assessment was conducted using the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2) tools recommended by the Cochrane Library. Result and conclusion This meta-analysis included a total of 38 studies of serum HE4 involving 14,745 subjects and five studies for urinary HE4 involving 649 subjects. We observed acceptable pooled sensitivity, specificity, summary receiver operating characteristics (SROC), and diagnostic odds ratio (DOR) at 0.79 (95% CI: 0.75-0.82), 0.92 (95% CI: 0.87-0.95), 0.88 (95% CI: 0.85-0.91), and 43 (95% CI: 25-72), respectively, for serum HE4 for discriminating ovarian cancer. For urine HE4, the pooled sensitivity, specificity, SROC, and DOR were 0.80 (95% CI: 0.64-0.90), 0.93 (95% CI: 0.83-0.98), 0.94 (95% CI: 0.91-0.95), and 55 (95% CI: 15-198), respectively. Therefore, HE4 is a promising biomarker with a high degree of specificity and acceptable sensitivity for the diagnosis of ovarian cancer. Registration number This meta-analysis was performed after the registration of the protocol in the PROSPERO database with registration number CRD42022324947.

Selective gas detection of titania nanoparticles via impedance spectroscopy and capacitive measurement.

The present paper demonstrated the impedance analysis of Au/TiO2nanoparticles/Si-Al capacitive sensor for selective detection of volatile organic compounds (VOCs) at different frequency regimes. TiO2nanoparticles (NP) were synthesized through the solution process and characterized by field-emission scanning electron microscopy , x-ray diffraction analysis, photoluminescence spectroscopy, and atomic force microscopy. The gas sensitivity of Au/TiO2-NP/Si-Al was investigated, with the effect of temperature modulation (25 °C-250 °C) and dielectric variation in the vicinity of nanoparticles. Impedance spectroscopy of TiO2-NP was carried out to obtain resonant peaks over the frequency ranging from 0.05 to 225 kHz and fitted with a complex nonlinear least-squares method. The optimum sensor response of 136%, 63%, 152%, and 174% was found at resonant frequencies of 0.38 kHz, 0.22 kHz, 0.15 kHz, and 0.1 kHz for the exposure of 2-propanol, acetone, ethanol, and methanol, respectively. The fastest response time and recovery time were found to be 32/21 s, 31.2/8 s, 32.5/9 s, and 40/26 s for acetone, 2-propanol, ethanol, and methanol, respectively. Selective detection of different VOCs at various resonant frequencies has correlated with the dielectric variation of the NPs and their associated void region under gas exposure.

Planning for resilience in screening operations using discrete event simulation modeling: example of HPV testing in Peru.

BACKGROUND: The World Health Organization (WHO) has called for the elimination of cervical cancer. Unfortunately, the implementation of cost-effective prevention and control strategies has faced significant barriers, such as insufficient guidance on best practices for resource and operations planning. Therefore, we demonstrate the value of discrete event simulation (DES) in implementation science research and practice, particularly to support the programmatic and operational planning for sustainable and resilient delivery of healthcare interventions. Our specific example shows how DES models can inform planning for scale-up and resilient operations of a new HPV-based screen and treat program in Iquitos, an Amazonian city of Peru. METHODS: Using data from a time and motion study and cervical cancer screening registry from Iquitos, Peru, we developed a DES model to conduct virtual experimentation with "what-if" scenarios that compare different workflow and processing strategies under resource constraints and disruptions to the screening system. RESULTS: Our simulations show how much the screening system's capacity can be increased at current resource levels, how much variability in service times can be tolerated, and the extent of resilience to disruptions such as curtailed resources. The simulations also identify the resources that would be required to scale up for larger target populations or increased resilience to disruptions, illustrating the key tradeoff between resilience and efficiency. Thus, our results demonstrate how DES models can inform specific resourcing decisions but can also highlight important tradeoffs and suggest general "rules" for resource and operational planning. CONCLUSIONS: Multilevel planning and implementation challenges are not unique to sustainable adoption of cervical cancer screening programs but represent common barriers to the successful scale-up of many preventative health interventions worldwide. DES represents a broadly applicable tool to address complex implementation challenges identified at the national, regional, and local levels across settings and health interventions-how to make effective and efficient operational and resourcing decisions to support program adaptation to local constraints and demands so that they are resilient to changing demands and more likely to be maintained with fidelity over time.

Transmembrane stem cell factor protein therapeutics enhance revascularization in ischemia without mast cell activation.

Stem cell factor (SCF) is a cytokine that regulates hematopoiesis and other biological processes. While clinical treatments using SCF would be highly beneficial, these have been limited by toxicity related to mast cell activation. Transmembrane SCF (tmSCF) has differential activity from soluble SCF and has not been explored as a therapeutic agent. We created novel therapeutics using tmSCF embedded in proteoliposomes or lipid nanodiscs. Mouse models of anaphylaxis and ischemia revealed the tmSCF-based therapies did not activate mast cells and improved the revascularization in the ischemic hind limb. Proteoliposomal tmSCF preferentially acted on endothelial cells to induce angiogenesis while tmSCF nanodiscs had greater activity in inducing stem cell mobilization and recruitment to the site of injury. The type of lipid nanocarrier used altered the relative cellular uptake pathways and signaling in a cell type dependent manner. Overall, we found that tmSCF-based therapies can provide therapeutic benefits without off target effects.

Spontaneous Esophageal Perforation in an Infant: A Rare Case.

Spontaneous perforation of the esophagus is an emergency that requires early diagnosis and management. It may be fatal and delay in treatment can cause an increase in morbidity and mortality. Despite of being very rare in infants, we have to be watchful whenever we encounter signs and symptoms related to it. Only 7 cases of spontaneous esophageal perforation in infants have been report in the literature to the best of our knowledge. Here we are reporting a rare case of spontaneous esophageal rupture in an infant.

Effective Downregulation of BCR-ABL Tumorigenicity by RNA Targeted CRISPR-Cas13a.

AIM: To induce BCR-ABL gene silencing using CRISPR Cas13a. BACKGROUND: CML is a clonal myeloproliferative disorder of pluripotent stem cells driven by a reciprocal translocation between chromosomes 9 and 22 forming a BCR-ABL fusion gene. Tyrosine- kinase inhibitor drugs like imatinib are the mainstay of treatment and cases resistant to these drugs have a poor prognosis in the absence of a compatible stem-cell donor. However with rapid advancements in gene-editing technologies most studies are now focusing on developing a translational model targeting single-gene disorders with a prospective permanent cure. OBJECTIVE: To explore the potential application of the RNA targeting CRISPR-Cas13a system for effective knockdown of BCR-ABL fusion transcript in a CML cell line K562. METHODS: CRISPR Cas13a crRNA was designed specific to the chimeric BCR-ABL gene and the system was transfected as a two-plasmid system into a CML cell line K562. The effects were enumerated by evaluating the expression levels of downstream genes dependent on the expression of the BCR-ABL gene. Also next-generation sequencing was used to ascertain the effects of CRISPR on the gene. RESULTS: The CRISPR system was successfully able to lower the expression of downstream genes [pCRKL and pCRK] dependent on the activated BCR-ABL kinase signal by up-to 4.3 folds. The viability of the CRISPR-treated cells was also significantly lowered by 373.83-fold [p-value= 0.000891196]. The time-dependent kinetics also highlighted the significant in-vitro suppressive activity to last up to 8 weeks [p-value: 0.025]. As per the cDNA sequencing data from the Oxford MinION next-generation sequencer the CRISPR treated cells show 62.37% suspected cleaved reads. CONCLUSION: These preliminary results highlight an excellent potential application of RNA targeting CRISPRs in Haematological neoplasms like CML and should pave the way for further research in this direction.

Low-dose ketamine as an adjuvant for pain control in a cancer patient: a case report.

Cancer patients often suffer from pain related problems such as under-treatment of pain, ineffective and persistent opioid administration as well as adverse opioid use outcomes. There is a growing need for non-opioid analgesic alternatives for patients undergoing treatment for obstinate pain. Ketamine is a fast-acting N-methyl-D-aspartate (NMDA) receptor antagonist that has been emerging as an effective medication for pain alleviation. While protocols have been established for the use of Low-Dose Ketamine (LDK) for post-operative pain, there is growing evidence for using LDK as a clinical alternative to opioids in a palliative care setting. This case study involves monitoring the efficacy of LDK treatment in combination with opioid analgesics in a cancer patient in a hospital setting. This is a very selected case of a patient with Metastatic Prostate Cancer (Gleason 9 Adenocarcinoma) where LDK was shown to be efficacious at reducing pain when opioids and standard pain medications were not satisfactory. While the study involved using a relatively novel pharmacological protocol and close patient monitoring, the patient reported a sustained reduction in pain level based on the Numerical Rating Scale for months after the termination of LDK infusions. Moreover, the treatment also resulted in a reduction of total opioid usage after the addition of LDK. Although additional research is needed to ascertain optimal dosing schedules and route of Ketamine, given these promising findings, Ketamine may be a useful option for improving the treatment of refractory pain in patients with cancer and a good tool in palliative medicine for treating neoplastic pain.

Epigenetic analysis reveals significant differential expression of miR-378C and miR-128-2-5p in a cohort of relapsed pediatric B-acute lymphoblastic leukemia cases.

INTRODUCTION AND OBJECTIVE: Epigenetic changes play a major role in mediating chemoresistance and relapse in pediatric ALL, and hence in current pilot study, we tried to identify DNA methylation, miRNA expression, and copy number variations (CNVs) in a cohort of relapse pediatric B-ALL cases. METHODOLOGY: DNA methylation, miRNA expression, and CNV analysis were performed in a total of 14, 16, and 18 cases as diagnosis-relapse samples. Briefly, DNA methylation was performed using Infinium HumanMethylation850 chip and data analyzed using RnBeads. miRNA was sequenced on illumina NextSeq500 platform for 20M 75bp SE reads and analyzed by DESeq2. CNVs were assessed by MLPA assay using the ALL P-335 probemix kit and analyzed by coffalyzer.net. RESULTS: On methylation analysis, oncogenes MYCN, MYB, and EGFR and tumor suppressor genes MDM4 & BCL11B were found differentially expressed as compared to controls (p-0.03). In addition, protooncogenes-AXL, HCK, MED12, and ETS2-were hypomethylated/overexpressed in 4 or more cases (P < .05). miRNA analysis revealed significant differential expression of miR-128-2-5p and miR-378C (p-4.4e-15 and p-6.4E-12) in relapse samples. CNV analysis revealed that frequency of good and intermediate/poor risk CNV profile at diagnosis was nearly equal (40% vs 60%). However, CDKN2A/2B and IKZF1 gene CNVs if present in initial diagnostic clone usually persisted in relapse clone. DISCUSSION AND CONCLUSION: Our pilot study highlights two miRNAs (miR-128-2-5p and miR-378C) as possible candidate biomarkers of relapsed B-ALL. However, these miRNAs and hypomethylated protooncogene signature noted in our data needs validation in a larger series of B-ALL.

Driving Force for the Complexation of Charged Polypeptides.

The phase separation of oppositely charged polyelectrolytes in solution is of current interest. In this work, we study the driving force for polyelectrolyte complexation using molecular dynamics simulations. We calculate the potential of mean force between poly(lysine) and poly(glutamate) oligomers using three different force fields, an atomistic force field and two coarse-grained force fields. There is qualitative agreement between all force fields, i.e., the sign and magnitude of the free energy and the nature of the driving force are similar, which suggests that the molecular nature of water does not play a significant role. For fully charged peptides, we find that the driving force for association is entropic in all cases when small ions either neutralize the poly ions, or are in excess. The removal of all counterions switches the driving force, making complexation energetic. This suggests that the entropy of complexation is dominated by the counterions. When only 6 residues of a 11-mer are charged, however, the driving force is energetic in the abscence of excess salt. The simulations shed insight into the mechanism of complex coacervation and the importance of realistic models for the polyions.

High-throughput screening against protein:protein interaction interfaces reveals anti-cancer therapeutics as potent modulators of the voltage-gated Na+ channel complex.

Multiple voltage-gated Na+ (Nav) channelopathies can be ascribed to subtle changes in the Nav macromolecular complex. Fibroblast growth factor 14 (FGF14) is a functionally relevant component of the Nav1.6 channel complex, a causative link to spinocerebellar ataxia 27 (SCA27) and an emerging risk factor for neuropsychiatric disorders. Yet, how this protein:channel complex is regulated in the cell is still poorly understood. To search for key cellular pathways upstream of the FGF14:Nav1.6 complex, we have developed, miniaturized and optimized an in-cell assay in 384-well plates by stably reconstituting the FGF14:Nav1.6 complex using the split-luciferase complementation assay. We then conducted a high-throughput screening (HTS) of 267 FDA-approved compounds targeting known mediators of cellular signaling. Of the 65 hits initially detected, 24 were excluded based on counter-screening and cellular toxicity. Based on target analysis, potency and dose-response relationships, 5 compounds were subsequently repurchased for validation and confirmed as hits. Among those, the tyrosine kinase inhibitor lestaurtinib was highest ranked, exhibiting submicromolar inhibition of FGF14:Nav1.6 assembly. While providing evidence for a robust in-cell HTS platform that can be adapted to search for any channelopathy-associated regulatory proteins, these results lay the potential groundwork for repurposing cancer drugs for neuropsychopharmacology.