Nickel-doped magnetic carbon aerogel derived from xanthan gum: a competent catalyst for the degradation of single and binary dye-based water pollutants.

Toxic organic dyes (colorants) are one of the main causes of water pollution that releases destructive effluents in the environment. To overcome this issue, a fundamental need to produce a novel, efficient catalyst for the degradation and mineralization of dye mixtures has arisen. The objective of this research is to develop an eminent Ni-doped magnetic carbon aerogel (Ni-MCA) catalyst using graft co-polymerization method having xanthan gum as backbone doped with Ni-magnetic nanoparticles (Ni-MNPs), that do not show agglomeration and easy to separate. The examination revealed that Ni-MCA provided exceptional magnetic characteristics (Ms = 52.75 emu/g) and potent catalytic activity for the degradation of mono- as well as binary-dye solutions of Congo red (CR) and methyl green (MG) dyes. The formation was verified by various characterization techniques such as FTIR, VSM, XRD, XPS, SEM, TEM, and EDX mapping. Interestingly, Ni-MCA shows faster result on anionic dye CR up to 97% with degradation rate of 5.647 × 10-1 min-1, and MG dye shows degradation of 95.7% with the degradation rate of 2.169 × 10-1 min-1, while dye mixture is showing 90% degradation with rate of 2.159 × 10-1 min-1.

Corrigendum: A single-institution experience with 177Lu RPT workflow improvements and qualifying the SPECT/CT imaging for dosimetry.

[This corrects the article DOI: 10.3389/fonc.2024.1331266.].

A single-institution experience with 177Lu RPT workflow improvements and qualifying the SPECT/CT imaging for dosimetry.

Background and purpose: Implementing any radiopharmaceutical therapy (RPT) program requires a comprehensive review of system readiness, appropriate workflows, and training to ensure safe and efficient treatment delivery. A quantitative assessment of the dose delivered to targets and organs at risk (OAR) using RPT is possible by correlating the absorbed doses with the delivered radioactivity. Integrating dosimetry into an established RPT program demands a thorough analysis of the necessary components and system fine-tuning. This study aims to report an optimized workflow for molecular radiation therapy using 177Lu with a primary focus on integrating patient-specific dosimetry into an established radiopharmaceutical program in a radiation oncology setting. Materials and methods: We comprehensively reviewed using the Plan-Do-Check-Act (PDCA) cycle, including efficacy and accuracy of delivery and all aspects of radiation safety of the RPT program. The GE Discovery SPECT/CT 670DR™ system was calibrated per MIM protocol for dose calculation on MIM SurePlan™ MRT software. Jaszcak Phantom with 15-20 mCi of 177Lu DOTATATE with 2.5 µM EDTA solution was used, with the main energy window defined as 208 keV ±10% (187.6 to 229.2 keV); the upper scatter energy window was set to 240 keV ±5% (228 to 252 keV), while the lower scatter energy window was 177.8 keV ±5% (168.9 to 186.7 keV). Volumetric quality control tests and adjustments were performed to ensure the correct alignment of the table, NM, and CT gantry on SPECT/CT. A comprehensive end-to-end (E2E) test was performed to ensure workflow, functionality, and quantitative dose accuracy. Results: Workflow improvements and checklists are presented after systematically analyzing over 400 administrations of 177Lu-based RPT. Injected activity to each sphere in the NEMA Phantom scan was quantified, and the MIM Sureplan MRT reconstruction images calculated activities within ±12% of the injected activity. Image alignment tests on the SPECT/CT showed a discrepancy of more than the maximum tolerance of 2.2 mm on any individual axis. As a result of servicing the machine and updating the VQC and COR corrections, the hybrid imaging system was adjusted to achieve an accuracy of <1 mm in all directions. Conclusion: Workflows and checklists, after analysis of system readiness and adequate training for staff and patients, are presented. Hardware and software components for patient-specific dosimetry are presented with a focus on hybrid image registration and correcting any errors that affect dosimetric quantification calculation. Moreover, this manuscript briefly overviews the necessary quality assurance requirements for converting diagnostic images into dosimetry measurement tools and integrating dosimetry for RPT based on 177Lu.

Surgically targeted radiation therapy (STaRT) for recurrent brain metastases: Initial clinical experience.

PURPOSE: This study evaluates the outcomes of recurrent brain metastasis treated with resection and brachytherapy using a novel Cesium-131 carrier, termed surgically targeted radiation therapy (STaRT), and compares them to the first course of external beam radiotherapy (EBRT). METHODS: Consecutive patients who underwent STaRT between August 2020 and June 2022 were included. All patients underwent maximal safe resection with pathologic confirmation of viable disease prior to STaRT to 60 Gy to a 5-mm depth from the surface of the resection cavity. Complications were assessed using CTCAE version 5.0. RESULTS: Ten patients with 12 recurrent brain metastases after EBRT (median 15.5 months, range: 4.9-44.7) met the inclusion criteria. The median BED10Gy90% and 95% were 132.2 Gy (113.9-265.1 Gy) and 116.0 Gy (96.8-250.6 Gy), respectively. The median maximum point dose BED10Gy for the target was 1076.0 Gy (range: 120.7-1478.3 Gy). The 6-month and 1-year local control rates were 66.7% and 33.3% for the prior EBRT course; these rates were 100% and 100% for STaRT, respectively (p < 0.001). At a median follow-up of 14.5 months, there was one instance of grade two radiation necrosis. Surgery-attributed complications were observed in two patients including pseudomeningocele and minor headache. CONCLUSIONS: STaRT with Cs-131 presents an alternative approach for operable recurrent brain metastases and was associated with superior local control than the first course of EBRT in this series. Our initial clinical experience shows that STaRT is associated with a high local control rate, modest surgical complication rate, and low radiation necrosis risk in the reirradiation setting.

Etiology of Meningoencephalitis in children aged less than 5 years.

BACKGROUND: The incidence of meningoencephalitis (ME) in India is poorly understood, and the exact etiological diagnosis is often not possible. This study was planned to elucidate the bacterial and viral etiological diagnosis of ME in children less than 5 years of age. MATERIAL AND METHODS: The present study was conducted in Virus Research and Diagnostic Laboratory (VRDL), Department of Microbiology, King George's Medical University, Lucknow, from July 2020 to June 2022. Serum, cerebrospinal fluid (CSF), and nose/throat swabs were collected from all the enrolled cases of meningoencephalitis in children below 5 years of age and tested for various etiological agents by ELISA and/or real-time PCR. RESULTS: Of 130 enrolled cases, 50 (38.5%) cases tested positive for one or more etiological agents. Etiological agents of ME detected were Japanese encephalitis virus (JEV) (8.46%), adenovirus (6.92%), influenza virus (5.38%), dengue virus (3.85%), Parvo B-19 virus (3.08%), Orientia tsutsugamushi (3.08%), Herpes Simplex Virus-1 (HSV-1) (1.54%), measles virus (1.54%), and Varicella-Zoster Virus (VZV) (1.54%). Rubella virus, Chikungunya virus (CHKV), Mumps virus, Enteroviruses, Parecho virus, John Cunningham virus (JC), BK virus, Nipah virus, Kyasanur Forest Disease virus (KFD), Chandipura virus, Herpes Simplex Virus (HSV-2), SARS CoV-2, N. Meningitides, and H. Influenzae were tested but not detected in any of the cases. CONCLUSION: We identified the etiological agents in 50/130 (38.5%) suspected ME cases in children less than 5 years of age, using molecular and ELISA-based diagnostic methods. The four most common pathogens detected were JEV, adenovirus, influenza virus, and dengue virus.

Biopolymeric nanoparticles based effective delivery of bioactive compounds toward the sustainable development of anticancerous therapeutics.

Nowadays, effective cancer therapy is a global concern, and recent advances in nanomedicine are crucial. Cancer is one of the major fatal diseases and a leading cause of death globally. Nanotechnology provides rapidly evolving delivery systems in science for treating diseases in a site-specific manner using natural bioactive compounds, which are gaining widespread attention. Nanotechnology combined with bioactives is a very appealing and relatively new area in cancer treatment. Natural bioactive compounds have the potential to be employed as a chemotherapeutic agent in the treatment of cancer, in addition to their nutritional benefits. Alginate, pullulan, cellulose, polylactic acid, chitosan, and other biopolymers have been effectively used in the delivery of therapeutics to a specific site. Because of their biodegradability, biopolymeric nanoparticles (BNPs) have received a lot of attention in the development of new anticancer drug delivery systems. Biopolymer-based nanoparticle systems can be made in a variety of ways. These systems have developed as a cost-effective and environmentally friendly solution to boost treatment efficacy. Effective drug delivery systems with improved availability, increased selectivity, and lower toxicity are needed. Recent research findings and current knowledge on the use of BNPs in the administration of bioactive chemicals in cancer therapy are summarized in this review.

Relationship between tobacco use, alcohol consumption and non-communicable diseases among women in India: evidence from National Family Health Survey-2015-16.

BACKGROUND: Based on an increased prevalence of diabetes, asthma and hypertension among women in reproductive age, understanding the risk factors of non-communicable diseases (NCDs) is crucial to inform policy and program interventions to address the problem. In this study, we empirically assessed the associations of behavioural factors such as alcohol consumption and tobacco use and a variety of socioeconomic characteristics with prevalence of NCDs in adult women. METHODS: The data were derived from the National Family Health Survey conducted in 2015-16. The effective sample size for the present paper was 699,686 women aged 15-49 years in India. Descriptive statistics along with bivariate analysis were conducted to find the preliminary results. Additionally, multivariable logistic regression analysis was conducted to find the relationship between NCDs and behavioural factors such as alcohol consumption and tobacco use. Moreover, population attributable risk was estimated in the present study. RESULTS: It was revealed that 15.9% of women had any of the NCDs. A proportion of 0.8% of women smoked tobacco whereas 5.5% of women consumed smokeless tobacco. Also, a proportion of 1.2% of women consumed alcohol in the current study. The odds of having NCDs among women who smoked tobacco, consumed smokeless tobacco and consume alcohol were 16, 8 and 20% significantly higher than the odds of having NCDs among women who did not smoke tobacco, consume smokeless tobacco and consume alcohol respectively. The population attributable risk of having NCDs was 1.8% (p < 0.001) for women who smoked, 0.8% (p < 0.001) for women who consumed smokeless tobacco and 2.2% (p < 0.001) for women who consumed alcohol. Besides, the odds of having NCDs among overweight and obese women were 2.25 and 3.60 times greater than the odds of having NCDs among women who were underweight. CONCLUSION: The findings revealed that smoking and using smokeless tobacco and alcohol consumption were risk factors of NCDs in women. The findings also alarm the focus of maternal and child health programs on NCDs' risk factors like maternal obesity, due to their adverse health consequences on their children too. Also, the coexistence of higher levels of tobacco use and alcohol consumption requires different strategies to address the vulnerability of women towards NCDs, including screening and early detection of NCDs especially among those who smoke or chew tobacco and consume alcohol.

Probing the metabolism of γ-glutamyl peptides in cyanobacteria via metabolite profiling and 13 C labeling.

Cyanobacteria are attractive model organisms for the study of photosynthesis and diurnal metabolism and as hosts for photoautotrophic production of chemicals. Exposure to bright light or environmental pollutants and a diurnal lifestyle of these prokaryotes may result in significant oxidative stress. Glutathione is a widely studied γ-glutamyl peptide that plays a key role in managing oxidative stress and detoxification of xenobiotics in cyanobacteria. The functional role and biosynthesis pathways of this tripeptide have been studied in detail in various phyla, including cyanobacteria. However, other γ-glutamyl peptides remain largely unexplored. We use an integrated approach to identify a number of γ-glutamyl peptides based on signature mass fragments and mass shifts in them in 13 C and 15 N enriched metabolite extracts. The newly identified compounds include γ-glutamyl dipeptides and derivatives of glutathione. Carbon backbones of the former turn over much faster than that of glutathione, suggesting that they follow a distinct biosynthesis pathway. Further, transients of isotopic 13 C enrichment show positional labeling in these peptides, which allows us to delineate the alternative biosynthesis pathways. Importantly, the amino acid of γ-glutamyl dipeptides shows much faster turnover compared to the glutamate moiety. The significant accumulation of γ-glutamyl dipeptides under slow-growth conditions combined with the results from dynamic 13 C labeling suggests that these compounds may act as reservoirs of amino acids in cyanobacteria.

Population attributable risk for multimorbidity among adult women in India: Do smoking tobacco, chewing tobacco and consuming alcohol make a difference?

BACKGROUND: The present study aims to estimate the prevalence and correlates of multimorbidity among women aged 15-49 years in India. Additionally, the population attributable risk for multi-morbidity in reference to those women who smoke tobacco, chew tobacco, and consume alcohol is estimated. METHODS: The data was derived from the National Family Health Survey which was conducted in 2015-16. The effective sample size for the present paper 699,686 women aged 15-49 years in India. Descriptive statistics along with bivariate analysis were used to do the preliminary analysis. Additionally, binary logistic regression analysis was used to fulfil the objectives. RESULTS: About 1.6% of women had multimorbidity in India. The prevalence of multimorbidity was high among women from southern region of India. Women who smoke tobacco, chew tobacco and consume alcohol had 87% [AOR: 1.87CI: 1.65, 2.10], 18% [AOR: 1.18; CI: 1.10, 1.26] and 18% [AOR: 1.18; CI: 1.04, 1.33] significantly higher likelihood to suffer from multi-morbidity than their counterparts respectively. Population Attributable Risk for women who smoke tobacco was 1.2% (p<0.001), chew tobacco was 0.2% (p<0.001) and it was 0.2% (p<0.001) among women who consumed alcohol. CONCLUSION: The findings indicate the important role of lifestyle and behavioural factors such as smoking and chewing tobacco and consuming alcohol in the prevalence of multimorbidity among adult Indian women. The subgroups identified as at increased risk in the present study can be targeted while making policies and health decisions and appropriate comorbidity management can be implemented.

Processing and Thermal Conductivity of Bulk Nanocrystalline Aluminum Nitride.

Producing bulk AlN with grain sizes in the nano regime and measuring its thermal conductivity is an important milestone in the development of materials for high energy optical applications. We present the synthesis and subsequent densification of nano-AlN powder to produce bulk nanocrystalline AlN. The nanopowder is synthesized by converting transition alumina (δ-Al2O3) with <40 nm grain size to AlN using a carbon free reduction/nitridation process. We consolidated the nano-AlN powder using current activated pressure assisted densification (CAPAD) and achieved a relative density of 98% at 1300 °C with average grain size, d¯~125 nm. By contrast, high quality commercially available AlN powder yields densities ~75% under the same CAPAD conditions. We used the 3-ω method to measure the thermal conductivity, κ of two nanocrystalline samples, 91% dense, d¯ = 110 nm and 99% dense, d¯ = 220 nm, respectively. The dense sample with 220 nm grains has a measured κ = 43 W/(m·K) at room temperature, which is relatively high for a nanocrystalline ceramic, but still low compared to single crystal and large grain sized polycrystalline AlN which can exceed 300 W/(m·K). The reduction in κ in both samples is understood as a combination of grain boundary scattering and porosity effects. We believe that these are finest d¯ reported in bulk dense AlN and is the first report of thermal conductivity for AlN with ≤220 nm grain size. The obtained κ values are higher than the vast majority of conventional optical materials, demonstrating the advantage of AlN for high-energy optical applications.

Amino acid derived biopolymers: Recent advances and biomedical applications.

Over the past few years, amino acids (AA) have emerged as promising biomaterials for the synthesis of functional polymers. Owing to the diversity of functional groups in amino acids, various polymerization methods may be used to make a wide range of well-defined functional amino-acid/peptide-based optically active polymers with varying polymer lengths, compositions, and designs. When incorporated with chirality and self-assembly, they offer a wide range of applications and are particularly appealing in the field of drug delivery, tissue engineering, and biosensing. There are several classes of these polymers that include polyamides (PA), polyesters (PE), poly(ester-amide)s (PEA)s, polyurethanes (PU)s, poly(depsipeptide)s (PDP)s, etc. They offer the ability to control functionality, conjugation, crosslinking, stimuli responsiveness, and tuneable mechanical/thermal properties. In this review, we present the recent advancements in the synthesis strategies for obtaining these amino acid-derived bio-macromolecules, their self-assembly properties, and the wealth of prevalent applications.

Case report of visual biofeedback-driven, magnetic resonance-guided single-fraction SABR in breath hold for early stage non-small-cell lung cancer.

Stereotactic ablative body radiation therapy (SABR) is a well-established alternative to surgery for early stage non-small-cell lung cancer (NSCLC). While SABR is typically delivered in 3 to 5 fractions, randomized trials have shown single-fraction SABR to be a reasonable alternative. We present the case of a 66-year-old male with history of cholangiocarcinoma who was subsequently diagnosed with peripheral early stage NSCLC and treated in mid-inspiration breath hold (BH) to 34 Gy in 1 fraction on a magnetic resonance (MR)-guided linear accelerator, with treatment delivery completed in 17 minutes. Visual biofeedback was utilized to maximize patient compliance with appropriate depth of inspiration BH and improve overall treatment delivery time efficiency. The benefits of single- vs multifraction SABR and unique advantages of MR guidance that are particularly well-suited for single-fraction SABR are reviewed.

Ablative 5-Fraction Stereotactic Magnetic Resonance-Guided Radiation Therapy With On-Table Adaptive Replanning and Elective Nodal Irradiation for Inoperable Pancreas Cancer.

PURPOSE: Radiation therapy dose escalation using stereotactic body radiation therapy may significantly improve both local control (LC) and overall survival (OS) for patients with inoperable pancreas cancer. However, ablative dose cannot be routinely offered because of the risk of causing severe injury to adjacent normal organs. Stereotactic magnetic resonance (MR)-guided adaptive radiation therapy (SMART) represents a novel technique that may achieve safe delivery of ablative dose and improve long-term outcomes. METHODS AND MATERIALS: We performed a single institution retrospective analysis of 35 consecutive pancreatic cancer patients treated with SMART in mid-inspiration breath hold on an MR-linear accelerator. Most had locally advanced disease (80%) and received induction chemotherapy (91.4%) for a median 3.9 months before stereotactic body radiation therapy. All were prescribed 5 fractions delivered in consecutive days to a median total dose of 50 Gy (BED10 100 Gy10), typically with a 120% to 130% hotspot. Elective nodal irradiation was delivered to 20 (57.1%) patients. No patient had fiducial markers placed and all were treated with continuous intrafraction MR visualization and automatic beam triggering. RESULTS: With median follow-up of 10.3 months from SMART, acute (2.9%) and late (2.9%) grade 3 toxicities were uncommon. One-year LC, distant metastasis-free survival, progression-free survival, cause-specific survival, and OS were 87.8%, 63.1%, 52.4%, 77.6%, and 58.9%, respectively. CONCLUSIONS: To our knowledge, this is the first report of 5-fraction pancreas SMART delivered on an MR-linear accelerator. We observed minimal severe treatment-related toxicity and encouraging early LC. Prospective confirmation of feasibility and long-term clinical outcomes of dose intensified SMART is warranted.

Comparison of Postoperative Pain Management after Third Molar Extraction with Two Anti-Inflammatory Drugs.

Background: The present study was aimed to investigate the pre-emptive analgesia attained with oral celecoxib compared with oral acetaminophen post mandibular third molar surgery. Methodology: A double-blinded, randomized, placebo-controlled clinical trial was performed in order to examine patients having a mandibular third molar for extraction under local anesthesia. The patients were randomized for receiving a preoperative oral dosage of celecoxib or acetaminophen as the predictor parameter. The primary outcome parameter was postoperative pain assessed through a visual analog scale (VAS) at different time points. The secondary outcome parameter was the quantity of postoperative analgesics taken in both groups. Statistical analyses included descriptive statistics, the t test, and the Pearson c2 test. Significance was set at P < .05. Results: 60 patients were divided randomly into either celecoxib receiving group or acetaminophen receiving group. The postoperative pain scores in the celecoxib group receiving were significantly lower than those in the acetaminophen receiving group at 4, 6, 8, and 12 hours (P = .078, P = .0012, P = .0211, and P = .011, respectively). The number of patients who didn't require any analgesics in the celecoxib receiving group was less than that in the acetaminophen receiving group (P =.0186). The average amount of rescue analgesic medication in the celecoxib group (0.6 0.8 dose) was significantly lower than that in the acetaminophen group (1.3 1.0 doses) (P = .002). The Kaplan-Meier curve (KM curve) was indicative of long-term survival of the patient's receiving celecoxib compared to those who did not receive any rescue analgesic medication (P = .0055). Conclusions: Celecoxib, as the study reflects, has a significant pre-emptive analgesic effect, thus helpful in reducing the usage of postoperative analgesics after removal of the third molar.

Polycyclic aromatic hydrocarbons (PAHs) and esophageal carcinoma in Handan-Xingtai district, North China: a preliminary study based on cancer risk assessment.

Extremely high risk of esophageal carcinoma (EC) occurs in Handan-Xingtai district of North China. In spite of various preventive measures and epidemiological investigations that have been conducted for years, incidence and mortality of EC are still in the highest level of China. The etiology of EC remains unclear in the region. Previous studies of our research group proposed that polycyclic aromatic hydrocarbons (PAHs) that derived from numerous coal gangue dumps and atmospheric particulates were major contaminants in these regions. In consideration of mutagenic, teratogenic, and carcinogenic characteristics of PAHs, the authors hypothesized that severe exposure to PAHs could preform as a causative factor for EC. Therefore, four data sets documented in our previous studies were employed in this paper. To quantitatively evaluate the carcinogenic risk imposed by sixteen priority PAHs, incremental lifetime cancer risks (ILCRs) via three exposure pathways were calculated. The results showed that total ILCRs for adult group ranged from 2.08E-05 to 8.63E-02, with an average of 2.00E-02. Total ILCRs for childhood group ranged from 1.09E-05 to 4.48E-02, with an average of 1.04E-02. Total ILCR value of 94% samples exceeded 10-4, indicating a particularly high carcinogenic risk to local residents. Furthermore, ingestion and dermal contact conducted as principal pathways of exposing to PAHs for each population group, rather than inhalation. It can be speculated that severely exposing to PAHs may be a pathogenesis of EC in Handan-Xingtai district. The rigorous supervise and governance are imperative to avoid severe exposure to PAHs that derived from coal gangue dumps.

Thermodynamic interference with bile acid demicelleization reduces systemic entry and injury during cholestasis.

Bile acids (BA), with their large hydrophobic steroid nucleus and polar groups are amphipathic molecules. In bile, these exist as micelles above their critical micellar concentration (CMC). In blood at low concentrations, these exist as monomers, initiating cellular signals. This micellar to monomer transition may involve complex thermodynamic interactions between bile salts alone or with phospholipids, i.e. mixed micelles and the aqueous environment. We therefore went on to test if therapeutically relevant changes in temperature could influence micellar behavior of bile salts, and in turn whether this affected the biological responses in cells, and in vivo. Sodium taurocholate (STC) belongs to a major class of bile salts. STC has a CMC in the 5-8 mM range and its infusion into the pancreatic duct is commonly used to study pancreatitis. We thus studied micellar breakdown of STC using isothermal titration calorimetry (ITC), dynamic light scattering and cryogenic transmission electron microscopy. Under conditions relevant to the in vivo environment (pH 7.4, Na 0.15 M), ITC showed STC to have a U shaped reduction in micellar breakdown between 37 °C and 15 °C with a nadir at 25 °C approaching ≈90% inhibition. This temperature dependence paralleled pancreatic acinar injury induced by monomeric STC. Mixed micelles of STC and 1-palmitoyl, 2-oleyl phosphatidylcholine, a phospholipid present in high proportions in bile, behaved similarly, with ≈75% reduction in micellar breakdown at 25 °C compared to 37 °C. In vivo pancreatic cooling to 25 °C reduced the increase in circulating BAs after infusion of 120 mM (5%) STC into the pancreatic duct, and duct ligation. Lower BA levels were associated with improved cardiac function, reduced myocardial damage, shock, lung injury and improved survival independent of pancreatic injury. Thus micellar breakdown of bile salts is essential for their entry into the systemic circulation, and thermodynamic interference with this may reduce their systemic entry and consequent injury during cholestasis, such as from biliary pancreatitis.

Genetic Heterogeneity of BRAF Fusion Kinases in Melanoma Affects Drug Responses.

BRAF fusions are detected in numerous neoplasms, but their clinical management remains unresolved. We identified six melanoma lines harboring BRAF fusions representative of the clinical cases reported in the literature. Their unexpected heterogeneous responses to RAF and MEK inhibitors could be categorized upon specific features of the fusion kinases. Higher expression level correlated with resistance, and fusion partners containing a dimerization domain promoted paradoxical activation of the mitogen-activated protein kinase (MAPK) pathway and hyperproliferation in response to first- and second-generation RAF inhibitors. By contrast, next-generation αC-IN/DFG-OUT RAF inhibitors blunted paradoxical activation across all lines and had their therapeutic efficacy further increased in vitro and in vivo by combination with MEK inhibitors, opening perspectives in the clinical management of tumors harboring BRAF fusions.

The E3 ubiquitin ligase RNF40 suppresses apoptosis in colorectal cancer cells.

BACKGROUND: Colorectal cancer (CRC) is the fourth leading cause of cancer-related deaths worldwide, and deciphering underlying molecular mechanism is essential. The loss of monoubiquitinated histone H2B (H2Bub1) was correlated with poor prognosis of CRC patients and, accordingly, H2Bub1 was suggested as a tumor-suppressive mark. Surprisingly, our previous work revealed that the H2B ubiquitin ligase RING finger protein 40 (RNF40) might exert tumor-promoting functions. Here, we investigated the effect of RNF40 loss on tumorigenic features of CRC cells and their survival in vitro. METHODS: We evaluated the effects of RNF40 depletion in several human CRC cell lines in vitro. To evaluate cell cycle progression, cells were stained with propidium iodide and analyzed by flow cytometry. In addition, to assess apoptosis rates, caspase 3/7 activity was assessed in a Celigo® S-based measurement and, additionally, an Annexin V assay was performed. Genomic occupancy of H2Bub1, H3K79me3, and H3K27ac was determined by chromatin immunoprecipitation. Transcriptome-wide effects of RNF40 loss were evaluated based on mRNA-seq results, qRT-PCR, and Western blot. To rescue apoptosis-related effects, cells were treated with Z-VAD-FMK. RESULTS: Human CRC cell lines displayed decreased cell numbers in vitro after RNF40 depletion. While the differences in confluence were not mediated by changes in cell cycle progression, we discovered highly increased apoptosis rates after RNF40 knockdown due to elevated caspase 3/7 activity. This effect can be explained by reduced mRNA levels of anti-apoptotic and upregulation of pro-apoptotic BCL2 family members. Moreover, the direct occupancy of the RNF40-mediated H2B monoubiquitination was observed in the transcribed region of anti-apoptotic genes. Caspase inhibition by Z-VAD-FMK treatment rescued apoptosis in RNF40-depleted cells. However, knockdown cells still displayed decreased tumorigenic features despite the absence of apoptosis. CONCLUSIONS: Our findings reveal that RNF40 is essential for maintaining tumorigenic features of CRC cells in vitro by controlling the expression of genes encoding central apoptotic regulators.

Multimodal Transgastric Local Pancreatic Hypothermia Reduces Severity of Acute Pancreatitis in Rats and Increases Survival.

BACKGROUND & AIMS: Acute pancreatitis (AP) of different etiologies is associated with the activation of different signaling pathways in pancreatic cells, posing challenges to the development of targeted therapies. We investigated whether local pancreatic hypothermia, without systemic hypothermia, could lessen the severity of AP induced by different methods in rats. METHODS: A urethane balloon with 2 polyurethane tubes was placed inside the stomach of rats. AP was induced in Wistar rats by the administration of cerulein or glyceryl tri-linoleate (GTL). Then, cold water was infused into the balloon to cool the pancreas. Pancreatic temperatures were selected based on those found to decrease acinar cell injury. An un-perfused balloon was used as a control. Pancreatic and rectal temperatures were monitored, and an infrared lamp or heating pad was used to avoid generalized hypothermia. We collected blood, pancreas, kidney, and lung tissues and analyzed them by histology, immunofluorescence, immunoblot, cytokine and chemokine magnetic bead, and DNA damage assays. The effect of hypothermia on signaling pathways initiated by cerulein and GTL was studied in acinar cells. RESULTS: Rats with pancreatic cooling developed less severe GTL-induced AP compared with rats that received the control balloon. In acinar cells, cooling decreased the lipolysis induced by GTL, increased the micellar form of its fatty acid, lowered the increase in cytosolic calcium, prevented the loss of mitochondrial membrane potential (by 70%-80%), and resulted in a 40%-50% decrease in the uptake of a fatty acid tracer. In rats with AP, cooling decreased pancreatic necrosis by 48%, decreased serum levels of cytokines and markers of cell damage, and decreased markers of lung and renal damage. Pancreatic cooling increased the proportions of rats surviving 6 hours after induction of AP (to 90%, from <10% of rats that received the control balloon). In rats with cerulein-induced AP, pancreatic cooling decreased pancreatic markers of apoptosis and inflammation. CONCLUSIONS: In rats with AP, transgastric local pancreatic hypothermia decreases pancreatic necrosis, apoptosis, inflammation, and markers of pancreatitis severity and increases survival.

Myc and the Tip60 chromatin remodeling complex control neuroblast maintenance and polarity in Drosophila.

Stem cells establish cortical polarity and divide asymmetrically to simultaneously maintain themselves and generate differentiating offspring cells. Several chromatin modifiers have been identified as stemness factors in mammalian pluripotent stem cells, but whether these factors control stem cell polarity and asymmetric division has not been investigated so far. We addressed this question in Drosophila neural stem cells called neuroblasts. We identified the Tip60 chromatin remodeling complex and its interaction partner Myc as regulators of genes required for neuroblast maintenance. Knockdown of Tip60 complex members results in loss of cortical polarity, symmetric neuroblast division, and premature differentiation through nuclear entry of the transcription factor Prospero. We found that aPKC is the key target gene of Myc and the Tip60 complex subunit Domino in regulating neuroblast polarity. Our transcriptome analysis further showed that Domino regulates the expression of mitotic spindle genes previously identified as direct Myc targets. Our findings reveal an evolutionarily conserved functional link between Myc, the Tip60 complex, and the molecular network controlling cell polarity and asymmetric cell division.