Proteomic Analysis of HepG2 Cells Reveals FAT10 and BAG2 Signaling Pathways Affected by a Protease Inhibitor from Tinospora cordifolia (Willd.) Hook. f. and Thoms Stem. Extract Among the Different Plant and Microbial Samples Analyzed.

Objectives: Dysregulation of proteolysis underlies diseases like cancer. Protease inhibitors (PIs) regulate many biological functions and hence have potential anticancer properties. With this background, the current study aimed to identify the PI from natural sources such as plants and microbes against trypsin (a protease), which was assayed against casein, using an ultraviolet spectrophotometer-based methodology. Materials and Methods: PI extracted from a few plants and microbial samples were screened for their PI activity against trypsin. The PI from the most promising source in our study, Tinospora cordifolia (Willd.) Hook. f. and Thoms. stem, was partially purified using ammonium sulfate precipitation followed by dialysis. The PI activity of the partially purified inhibitor was analyzed against chymotrypsin and collagenase enzymes, and the cytotoxic effect of the PI was checked on HepG2 (liver carcinoma) cells by MTT- [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide]- assay. Liquid Chromatograography Mass Spectrometry -based proteomic studies were performed on HepG2 cells to understand the signaling pathways affected by the PIs in the liver cancer cell line. Results: Among the samples tested the PIs from T. cordifolia stem extract had the highest inhibitory activity (72.0%) against trypsin along with cytotoxicity to HepG2 cells. After partial purification by 80.0% ammonium sulfate precipitation, PI had increased inhibitory activity (83.0%) against trypsin and enhanced cytotoxicity (47.0%) to HepG2 cells. Proteomic analysis of the PI-treated HepG2 cells revealed that BAG2 and FAT10 signaling pathways were affected, which may have caused the inhibition of cancer cell proliferation. Conclusion: PI from T. cordifolia stem has promising anticancer potential and hence can be used for further purification and characterization studies toward cancer drug development.

A Comparative Study of Elective Sublay Versus Onlay Repair for Non-complex, Small, and Medium-Size Incisional Hernia: Post-operative Complications in a Tertiary Hospital in Ranchi, India.

BACKGROUND: The most difficult hernia surgery is the repair of the ventral hernia, which is caused by aberrant organ or tissue protrusions through the abdominal wall. Factors like obesity, smoking, and chronic medical conditions contribute to their formation. Surgical strategies have evolved from anatomical repair to mesh hernioplasty, with mesh placement playing a significant role in outcomes. The ideal anatomical location for mesh placement remains debated due to varying results. So, the objective of the study is to compare early postoperative complications, surgical site infection, and incidence of recurrence between sublay and onlay mesh placement repair of incisional hernias of <10 cm in diameter, at a tertiary hospital in Ranchi. METHODS:  This retrospective comparative study was conducted over a period of January 2022 to January 2024 at the Rajendra Institute of Medical Science, Ranchi, India. During the study period, 96 patients were operated on, and their demographic details, along with their position of mesh placement and postoperative complications (seroma formation, wound infection, postoperative hospital stays, and recurrence), were retrieved from the hospital data. Comparisons between onlay and sublay groups in terms of post-operative complications were made. RESULTS: Within the study period, a total of 96 patients were operated on for incisional hernia. In this study, 36 (37.5%) were male and 60 (62.5%) were female, with a male-to-female ratio of 0.6:1. Out of the total number of patients, 56 (58.4%) had a past history of emergency surgery. It was observed that there was a higher incidence of seroma formation in the onlay group compared to the sublay with a statistical significance p-value of 0.027. The incidence of wound infection was found to be statistically significant (p-value = 0.035) between the onlay and sublay groups. In a period of six-month follow-up, three patients of the total study population had an incidence of recurrent incisional hernia, of which two from the onlay group and one from the sublay group were present, and there was no statistical significance (p-value > 0.5). CONCLUSIONS: Based on our retrospective analysis, we can say that there is a lower incidence of postoperative complications and recurrence in sublay repair, along with a shorter postoperative hospital stay, making it a preferred method of repair over onlay.

Effectiveness of Mannheim's Peritonitis Index in Patients With Peritonitis Secondary to Hollow Viscus Perforation in a Tertiary Care Hospital in Jharkhand, India.

Introduction Peritonitis refers to the inflammation of the peritoneum and peritoneal cavity. Causes of peritonitis can be bacterial (gastrointestinal or non-gastrointestinal), chemical, traumatic, or ischemic. Peritonitis can be localized or diffuse, acute or chronic. Peritonitis can be primary, secondary, or tertiary, according to the pathogenesis. Peritonitis developed secondary to hollow viscus perforation is a life-threatening condition and a common cause of emergency surgery in India. The Mannheim peritonitis index (MPI) is a simple scoring system that can accurately predict the outcome of peritonitis. This study aimed to evaluate the effectiveness of MPI in predicting mortality risk or prognosis in patients with peritonitis due to hollow viscus perforation. Materials and methods This observational cross-sectional study at the Department of General Surgery, Rajendra Institute of Medical Sciences, Ranchi, involved 111 patients with peritonitis due to hollow viscus perforation from December 2021 to March 2022. Detailed history, clinical examination, relevant blood tests, and radiological investigations established a diagnosis of perforation peritonitis, followed by a score assessment. Data were analyzed using SPSS software (IBM Corp., Armonk, NY, USA). Results Patients >50 years had higher mortality (i.e., 18/43) than patients <50 years (i.e., 13/68). Overall mortality was 31, which included one in low risk, 12 in intermediate risk, and 18 in the high-risk group. Mortality was lowest in the low-risk group (i.e., 1/30), highest in the high-risk group (i.e., 18/40), and 12/41 in the intermediate-risk group; the p-value was <0.05, which was highly significant. Mortality was higher in patients presenting after 24 hours, having organ failure, and non-colonic sepsis. Conclusion The MPI scoring system is simple, easy to calculate, cost-effective, precise, and effective in assessing mortality and morbidity risk in patients with peritonitis due to hollow viscus perforation. It can also guide further management strategies.

Multi-cancer risk stratification based on national health data: a retrospective modelling and validation study.

BACKGROUND: Health care is experiencing a drive towards digitisation, and many countries are implementing national health data resources. Although a range of cancer risk models exists, the utility on a population level for risk stratification across cancer types has not been fully explored. We aimed to close this gap by evaluating pan-cancer risk models built on electronic health records across the Danish population with validation in the UK Biobank. METHODS: In this retrospective modelling and validation study, data for model development and internal validation were derived from the following Danish health registries: the Central Person Registry, the Danish National Patient Registry, the death registry, the cancer registry, and full-text medical records from secondary care records in the capital region. The development data included adults aged 16-86 years without previous malignant cancers in the time period from Jan 1, 1995, to Dec 31, 2014. The internal validation period was from Jan 1, 2015, to April 10, 2018, and the data included all adults without a previous indication of cancer aged 16-75 years on Dec 31, 2014. The external validation cohort from the UK Biobank included all adults without a previous indication of cancer aged 50-75 years. We used time-dependent Bayesian Cox hazard models built on the combined medical history of Danish individuals. A set of 1392 covariates from available clinical disease trajectories, text-mined basic health factors, and family histories were used to train predictive models of 20 major cancer types. The models were validated on cancer incidence between 2015 and 2018 across Denmark and on individuals in the UK Biobank. The primary outcomes were discrimination and calibration performance. FINDINGS: From the Danish registries, we included 6 732 553 individuals covering 60 million hospital visits, 90 million diagnoses, and a total of 193 million life-years between Jan 1, 1978, and April 10, 2018. Danish registry data covering the period from Jan 1, 2015, to April 10, 2018, were used to internally validate risk models, containing a total of 4 248 491 individuals who remained at risk of a primary malignant cancer diagnosis and 67 401 cancer cases recorded. For the external validation, we evaluated the same time period in the UK Biobank covering 377 004 individuals with 11 486 cancer cases. The predictive performance of the models on Danish data showed good discrimination (concordance index 0·81 [SD 0·08], ranging from 0·66 [95% CI 0·65-0·67] for cervix uteri cancer to 0·91 [0·90-0·92] for liver cancer). Performance was similar on the UK Biobank in a direct transfer when controlling for shifts in the age distribution (concordance index 0·66 [SD 0·08], ranging from 0·55 [95% CI 0·44-0·66] for cervix uteri cancer to 0·78 [0·77-0·79] for lung cancer). Cancer risks were associated, in addition to heritable components, with a broad range of preceding diagnoses and health factors. The best overall performance was seen for cancers of the digestive system (oesophageal, stomach, colorectal, liver, and pancreatic) but also thyroid, kidney, and uterine cancers. INTERPRETATION: Data available in national electronic health databases can be used to approximate cancer risk factors and enable risk predictions in most cancer types. Model predictions generalise between the Danish and UK health-care systems. With the emergence of multi-cancer early detection tests, electronic health record-based risk models could supplement screening efforts. FUNDING: Novo Nordisk Foundation and the Danish Innovation Foundation.

Ultrasonication-assisted synthesis of transition metal carbide of MXene: an efficient and promising material for photocatalytic organic dyes degradation of rhodamine B and methylene blue in wastewater.

Water pollutants of non-biodegradable toxic aromatic dye including Methylene blue (MB) and Rhodamine (RhB) are extremely carcinogenic thiazines used in various industries such as leather industry, paper industry, and the dyeing industry. The presence of dyes in wastewater causes severe threats to human health that are responsible for various harmful chronic or acute diseases and also shows an adverse impact on the environment as it reduces transparency and is harmful to water microorganisms. To overcome severe issues, many traditional techniques have been used to remove toxic pollutants, but these methods are insufficient to remove chemically stable dyes that remain in the treated wastewater. However, the photocatalytic degradation process is an efficient approach to degrade the dye up to the maximum extent with improved efficiency. Therefore, in this work, a new class of two-dimensional (2D) transition metal carbide of Titanium Carbide (Ti3C2Tx) MXene material was used for the organic dyes degradation such as MB and RhB using a photocatalytic process. A layered structure of hexagonal lattice symmetry of Ti3C2Tx MXene was successfully synthesized from the Titanium Aluminum Carbide of Ti3AlC2 bulk phase using an exfoliation process. Further, the XRD spectrum confirms the transformation of bulk MAX phase having (002) plane at 9.2° to Ti3C2Tx MXene of (002) plane at 8.88° confirms the successful removal of Al layer from MAX phase. A smooth, transparent, thin sheet-like morphology of Ti3C2Tx nanosheet size were found to be in the range of 70 to 150 nm evaluated from TEM images. Also, no holes or damages in the thin sheets were found after the treatment with strong hydrofluoric acid confirms the formation Ti3C2Tx layered sheets. The synthesized Ti3C2Tx MXene possesses excellent photocatalytic activity for the degradation of dyes MB, RhB, and mixtures of MB and RhB dyes. MB dye degraded with a degradation percentage efficiency of 99.32% in 30 min, while RhB dye was degraded upto 98.9% in 30 min. Also, experiments were conducted for degradation of mixture of MB and RhB dyes by UV light, and the degradation percentage efficiency were found to be 98.9% and 99.75% for mixture of MB and RhB dye in 45 min, respectively. Moreover, reaction rate constant (k) was determined for each dye of MB, RhB, and mixtures of MB and RhB and was found to be 0.0215 min-1 and 0.0058 min-1, and for mixtures, it was 0.0020 min-1 and 0.009 min-1, respectively.

Combination of EZH2 and ATM inhibition in BAP1-deficient mesothelioma.

BACKGROUND: More than half of mesothelioma tumours show alterations in the tumour suppressor gene BAP1. BAP1-deficient mesothelioma is shown to be sensitive to EZH2 inhibition in preclinical settings but only showed modest efficacy in clinical trial. Adding a second inhibitor could potentially elevate EZH2i treatment efficacy while preventing acquired resistance at the same time. METHODS: A focused drug synergy screen consisting of 20 drugs was performed by combining EZH2 inhibition with a panel of anti-cancer compounds in mesothelioma cell lines. The compounds used are under preclinical investigation or already used in the clinic. The synergistic potential of the combinations was assessed by using the Bliss model. To validate our findings, in vivo xenograft experiments were performed. RESULTS: Combining EZH2i with ATMi was found to have synergistic potential against BAP1-deficient mesothelioma in our drug screen, which was validated in clonogenicity assays. Tumour growth inhibition potential was significantly increased in BAP1-deficient xenografts. In addition, we observe lower ATM levels upon depletion of BAP1 and hypothesise that this might be mediated by E2F1. CONCLUSIONS: We demonstrated the efficacy of the combination of ATM and EZH2 inhibition against BAP1-deficient mesothelioma in preclinical models, indicating the potential of this combination as a novel treatment modality using BAP1 as a biomarker.

Immunometabolic Regulation of Bacterial Infection, Biofilms, and Antibiotic Susceptibility.

BACKGROUND: Upon infection, mucosal tissues activate a brisk inflammatory response to clear the pathogen, i.e., resistance to disease. Resistance to disease is orchestrated by tissue-resident macrophages, which undergo profound metabolic reprogramming after sensing the pathogen. These metabolically activated macrophages release many inflammatory factors, which promote their bactericidal function. However, in immunocompetent individuals, pathogens like Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella evade this type of immunity, generating communities that thrive for the long term. SUMMARY: These organisms develop features that render them less susceptible to eradication, such as biofilms and increased tolerance to antibiotics. Furthermore, after antibiotic therapy withdrawal, "persister" cells rapidly upsurge, triggering inflammatory relapses that worsen host health. How these pathogens persisted in inflamed tissues replete with activated macrophages remains poorly understood. KEY MESSAGES: In this review, we discuss recent findings indicating that the ability of P. aeruginosa, S. aureus, and Salmonella to evolve biofilms and antibiotic tolerance is promoted by the similar metabolic routes that regulate macrophage metabolic reprogramming.

Methylglyoxal metabolism is altered during defence response in pigeonpea (Cajanus cajan (L.) Millsp.) against the spotted pod borer (Maruca vitrata).

Pigeonpea (Cajanus cajan ) production can be affected by the spotted pod borer (Maruca vitrata ). Here, we identified biochemical changes in plant parts of pigeonpea after M. vitrata infestation. Two pigeonpea genotypes (AL 1747, moderately resistant; and MN 1, susceptible) were compared for glyoxalase and non-glyoxalase enzyme systems responsible for methylglyoxal (MG) detoxification, γ-glutamylcysteine synthetase (γ-GCS), glutathione-S-transferase (GST) and glutathione content in leaves, flowers and pods under control and insect-infested conditions. MN 1 had major damage due to M. vitrata infestation compared to AL 1747. Lower accumulation of MG in AL 1747 was due to higher activities of enzymes of GSH-dependent (glyoxylase I, glyoxylase II), GSH-independent (glyoxalase III) pathway, and enzyme of non-glyoxalase pathway (methylglyoxal reductase, MGR), which convert MG to lactate. Decreased glyoxylase enzymes and MGR activities in MN 1 resulted in higher accumulation of MG. Higher lactate dehydrogenase (LDH) activity in AL 1747 indicates utilisation of MG detoxification pathway. Higher glutathione content in AL 1747 genotype might be responsible for efficient working of MG detoxification pathway under insect infestation. Higher activity of γ-GCS in AL 1747 maintains the glutathione pool, necessary for the functioning of glyoxylase pathway to carry out the detoxification of MG. Higher activities of GST and GPX in AL 1747 might be responsible for detoxification of toxic products that accumulates following insect infestation, and elevated activities of glyoxylase and non-glyoxylase enzyme systems in AL 1747 after infestation might be responsible for reducing reactive cabanoyl stress. Our investigation will help the future development of resistant cultivars.

Combined Inhibition of EZH2 and FGFR is Synergistic in BAP1-deficient Malignant Mesothelioma.

Malignant mesothelioma is a highly aggressive tumor with a survival of only 4-18 months after diagnosis. Treatment options for this disease are limited. Immune checkpoint blockade using ipilimumab and nivolumab has recently been approved as a frontline therapy, but this led to only a small improvement in overall patient survival. As more than half of patients with mesothelioma have alterations in the gene encoding for BAP1 this could be a potential marker for targeted therapies. In this study, we investigated the synergistic potential of combining EZH2 inhibition together with FGFR inhibition for treatment of BAP1-deficient malignancies. The efficacy of the combination was evaluated using human and murine preclinical models of mesothelioma and uveal melanoma in vitro. The efficacy of the combination was further validated in vivo by using BAP1-deficient mesothelioma xenografts and autochthonous mouse models. In vitro data showed sensitivity to the combined inhibition in BAP1-deficient mesothelioma and uveal melanoma tumor cell lines but not for BAP1-proficient subtypes. In vivo data showed susceptibility to the combination of BAP1-deficient xenografts and demonstrated an increase of survival in autochthonous models of mesothelioma. These results highlight the potential of this novel drug combination for the treatment of mesothelioma using BAP1 as a biomarker. Given these encouraging preclinical results, it will be important to clinically explore dual EZH2/FGFR inhibition in patients with BAP1-deficient malignant mesothelioma and justify further exploration in other BAP1 loss-associated tumors. SIGNIFICANCE: Despite the recent approval of immunotherapy, malignant mesothelioma has limited treatment options and poor prognosis. Here, we observe that EZH2 inhibitors dramatically enhance the efficacy of FGFR inhibition, sensitising BAP1-mutant mesothelioma and uveal melanoma cells. The striking synergy of EZH2 and FGFR inhibition supports clinical investigations for BAP1-mutant tumors.

Clinical and Demographic Characteristics of Patients with Coexistent Hypertension, Type 2 Diabetes Mellitus, and Dyslipidemia: A Retrospective Study from India.

BACKGROUND: Coexisting hypertension, type 2 diabetes mellitus (T2DM), and dyslipidemia (triple disease) can lead to greater risk of cardiovascular morbidity and mortality. The present study sought to comprehend the prevalence, demographic traits, clinical traits, and treatment patterns in Indian patients with these coexisting conditions. METHODS: An electronic medical record (EMR)-based, retrospective, multicenter, cross-sectional study was conducted, and data were collected for patients who were diagnosed with coexistent hypertension, T2DM, and dyslipidemia. Baseline patient variables evaluated were the percentage of patients with triple comorbidity, demographic characteristics, diagnostic laboratory parameters, and treatment pattern details. RESULTS: Data from 4793 centers (clinics) were included, with a total of 6,722,173 patients. Of these, 427,835 (6.36%) patients were found to have coexistent hypertension, T2DM, and dyslipidemia. Most of the patients belonged to the 40-64 year age group (62.10%) and were males (57.00%), while 27.40% patients had a body mass index (BMI) within normal limits, 43.30% patients were pre-obese, and 20.90% patients were class 1 obese. Further, 3402 patients (0.80%) had a recorded history of smoking. Mean glycated hemoglobin (HbA1c) for the patients included in the study was 8.35 ± 1.96 g%. Mean systolic blood pressure (SBP) was 138.81 ± 19.59 mm Hg, while mean diastolic blood pressure (DBP) was 82.17 ± 10.35 mm Hg; 27.60% cases had SBP < 130 mm Hg, while 28.37% cases had DBP < 80 mm Hg. The mean low-density lipoprotein (LDL), total cholesterol, and high-density lipoprotein (HDL) in mg/dl were 98.38 ± 40.39, 174.75 ± 46.73, and 44.5 ± 10.05, respectively. Of the enrolled cases, 55.64% had serum LDL below 100 mg/dl, 72.03% cases had serum cholesterol below 200 mg/dl, and 44.15% males and 71.77% females had serum HDL below the normal prescribed range. The most common monotherapy used for managing hypertension was angiotensin receptor blockers (ARB) (24.80%), followed by beta-blockers (24.30%). The most common combinations administered for management of hypertension were antihypertensives with diuretics (14.30%), followed by ARB plus calcium channel blockers (CCB) (13.30%). For dyslipidemia, the majority of patients (56.60%) received lipid-lowering medication in combination with drugs for other comorbidities. The most common antidiabetic agents prescribed were biguanides (74.60%). CONCLUSIONS: Coexistence of triple disease is not uncommon in the Indian population, with middle-aged patients diagnosed as pre-obese and obese being affected more commonly and receiving treatment for the same. The present study highlights that, though there are medications against the three chronic conditions, the rate of uncontrolled cases of hypertension, T2DM, and dyslipidemia remains high. Coexistence of triple disease increases the risk of cardiovascular and renal complications, which need to be closely monitored and effectively treated.

Incidental Identification of Osteoid Osteoma of Skull Bone on 68 Ga-PSMA PET/CT.

ABSTRACT: An osteoid osteoma (OO) is a benign bone neoplasm, characterized by significant nocturnal pain that usually responds to nonsteroidal anti-inflammatory drugs. It occurs most commonly in the lower extremities and vertebrae. Here, we present a case of carcinoma prostate, who was referred to our department for 68 Ga-PSMA PET/CT scan, and we incidentally found out PSMA-avid OO involving frontal bone of skull, which is a rare finding. To the best of our knowledge, this is the second case in which high PSMA uptake is found in the OO, suggesting a possible PSMA expression related to osteoblastic activity.

A Clinicopathological Study of OSMF to Evaluate the Response After Treatment with Intralesional Steroid Versus Intralesional Steroid Plus Hyaluronidase.

Oral submucous fibrosis (OSMF) is a chronic debilitating and a well-recognized potentially malignant condition of the oral cavity, sometimes involving oropharynx associated with trismus and burning sensation. Apart from medical management and counselling, local injection of hyaluronidase mixed with triamcinolone acetonide has been used at our centre for the last 20 years with satisfactory clinical results and without any significant side effects. The problem with the treatment was that the doses and duration of treatment has not been standardized. Therefore, in this study, authors aim to evaluate and compare the efficacy of Triamcinolone alone versus Triamcinolone acetonide plus Hyaluronidase at weekly interval and improvement in Clinical and Histopathological staging of disease after 6 weeks of treatment. This study was conducted in Department of Otorhinolaryngology & Head Neck Surgery, with a total sample of 80 participants divided into two Groups, group A received Inj. Triamcinolone acetonide and group B received Inj. Triamcinolone Acetonide and Hyaluronidase 1500 IU at weekly interval. Pre-treatment and post-treatment clinical and histopathological profile of the patients were recorded and analyzed using SPSS 16 software. According to pre-treatment status, the proportion of clinical grades I, II and III were found in proportion 12.5%, 18.8% and 15.0% respectively. No significant difference was found in proportion of various grades between the groups (p = 0.388). At post treatment, the grading was reduced with changed proportion of grades I, II and III cases as 33.8%, 41.3% and 7.5% respectively. There was no significant difference in proportion of various grades between the groups (p = 0.681). Further, the intragroup comparison showed significant improvement Pre to post in group A (p = 0.002), Group B (p < 0.001) and overall, as well (p < 0.001). The inj. Triamcinolone acetonide and Inj. Hyaluronidase showed a better improvement on post treatment histopathological grading although the difference between the two groups was not significant statistically.

Peroneal Artery Pseudoaneurysm after Surgery for Non-union of Tibia: A Case Report with Review of Literature.

Introduction: Arterial pseudoaneurysm is a hematoma that is formed after damage to the arterial wall. We report a rare case of peroneal artery pseudoaneurysm after open reduction and internal fixation with interlocking nailing and partial fibulectomy for non-union for the right tibia in a 31-year-old male. The patient presented with a bleeding sinus over the leg swelling, and it was managed with an exploration of the pseudoaneurysm and ligation of the peroneal artery. Case Report: A 30-year-old male patient presented with a non-union tibia on the right side and had undergone plating of the tibia at another institute for a fracture of both bone legs approximately 18 months ago. The revision surgery was performed in which a previously inserted implant was removed and an interlocking nail was inserted, along with a partial fibulectomy. The post-operative period was uneventful. At 8 weeks after the second surgery, the patient came with a complaint of swelling at the outer aspect of the right leg. Computed tomography and angiography confirmed a peroneal artery pseudoaneurysm of 3.2 × 2.8 × 3.8 cm. Pseudoaneurysm was explored, and the artery was overrun with a Figure-8 stitches using a monofilamentous, and non-absorbable suture. Conclusion: This case report highlights the occurrence of pseudoaneurysm after an orthoapedic procedure such as a partial fibulectomy. A high level of clinical suspicion, proper imaging, and early endovascular or surgical intervention is recommended to prevent complications.

G9a and Sirtuin6 epigenetically modulate host cholesterol accumulation to facilitate mycobacterial survival.

Cholesterol derived from the host milieu forms a critical factor for mycobacterial pathogenesis. However, the molecular circuitry co-opted by Mycobacterium tuberculosis (Mtb) to accumulate cholesterol in host cells remains obscure. Here, we report that the coordinated action of WNT-responsive histone modifiers G9a (H3K9 methyltransferase) and SIRT6 (H3K9 deacetylase) orchestrate cholesterol build-up in in vitro and in vivo mouse models of Mtb infection. Mechanistically, G9a, along with SREBP2, drives the expression of cholesterol biosynthesis and uptake genes; while SIRT6 along with G9a represses the genes involved in cholesterol efflux. The accumulated cholesterol in Mtb infected macrophages promotes the expression of antioxidant genes leading to reduced oxidative stress, thereby supporting Mtb survival. In corroboration, loss-of-function of G9a in vitro and pharmacological inhibition in vivo; or utilization of BMDMs derived from Sirt6-/- mice or in vivo infection in haplo-insufficient Sirt6-/+ mice; hampered host cholesterol accumulation and restricted Mtb burden. These findings shed light on the novel roles of G9a and SIRT6 during Mtb infection and highlight the previously unknown contribution of host cholesterol in potentiating anti-oxidative responses for aiding Mtb survival.

A Cross-Sectional Survey on the Use of Tobacco, Attempts on Cessation, and Locus of Control Among College Students.

Background Socio-cultural considerations (such as drug availability) and psychological traits play a significant role in predicting whether a person will use drugs in the future and dependency on the drugs. Second-, third-, and fourth-hand smoking and E-cigarettes are influencing factors for the use of tobacco in college students. This study conducted research to ascertain whether there is a potential relationship between tobacco consumption and various factors, including internal and external control sites and demographics. Materials and methods Participants in the study were found by walk-up distribution at multiple campus-wide smoking places, department announcements, and on-campus advertisements. Social media and participant references were also used in this study as recruitment tools. In addition, the locus of control questionnaire also identifies if the participating individual had extrinsic or intrinsic reinforcing routines. The classification of the participating individuals into respective internal and external locus of control was in accordance with their response survey after which a statistical analysis was done. Results This study found an association between smoking on campus and reported attempts to quit. Additionally, there is a strong association (r(85) = 0.31, p < 0.01) between reported tobacco use status and cigarette use on campus. Participants' gender and smoking status also had r(85) = 0.39, p-value < 0.01 correlation. The bulk of respondents indicated that they were seniors and off-campus living concluding for 36% (n = 34) and 60% (n = 51) of the total. Twenty-seven percent (n = 24) of the respondents were first-year college students and the rest 33% (n = 29) said their parents had no college education at all or incomplete college education. Conclusion Whenever there is a strong perception of organizational support for anti-tobacco policies, and improving compliance, there is a drastic increase in cigarette cessation and a drop in tobacco usage among those who still smoke. Perceived organizational support is strongly and positively connected with cessation among the organization's members.

The Value of the Sequential Organ Failure Assessment (SOFA) Score and Serum Lactate Level in Sepsis and Its Use in Predicting Mortality.

Background and objective Sepsis is a major health burden that leads to significant morbidity and mortality. Early diagnosis and severity prediction using various scoring systems can reduce the mortality rate, particularly in developing nations. There are two aims of this study. One is to evaluate the prognostic accuracy of the Sequential Organ Failure Assessment (SOFA) score and serum lactate levels in patients with sepsis to predict mortality. The other aim is to evaluate the relationship between the SOFA score and lactate so that we may be able to use lactate as a surrogate predictor of organ dysfunction and mortality in sepsis. Methods An observational prognostic accuracy study was conducted in the Department of General Surgery, Intensive Care Unit (ICU), Rajendra Institute of Medical Sciences (RIMS), Ranchi, Jharkhand, India, between 1 July 2021 and 1 October 2022. We selected 128 patients, calculated their SOFA and lactate levels, and divided them into survivors and non-survivors according to their outcomes after seven days of assessment. The SOFA score and serum lactate levels were assessed as predictors of mortality, and their correlation was studied. Results We observed a significant decreasing trend in the value of the mean SOFA, maximum SOFA, mean lactate, and maximum lactate among survivors, whereas an increasing trend for the same was observed in non-survivors. The receiver operating characteristic (ROC) analysis showed the best diagnostic accuracy of the mean lactate (area under the curve {AUC}=0.996, 95% confidence interval {CI}=0.964-1.00, p≤0.0001). The maximum lactate (AUC=0.987, 95% CI=0.949-0.999, p≤0.0001) and mean SOFA scores (AUC=0.986, 95% CI=0.948-0.999, p≤0.0001) were good at predicting the mortality in sepsis. A slightly lower diagnostic accuracy was found for the maximum SOFA score (AUC=0.969, 95% CI=0.923-0.992, p≤0.0001). There was a strong correlation between the mean lactate and the mean SOFA with a correlation coefficient of 0.883 and p=0.0001. A good correlation was found between maximum lactate and maximum SOFA too (correlation coefficient=0.873, p≤0.0001). Conclusion This study highlights the different predictors of mortality in the patients with sepsis. The maximum lactate was the most accurate in predicting mortality in sepsis. It also demonstrates how serum lactate, due to its strong correlation with the SOFA score, can be used in its place to predict mortality in sepsis and organ dysfunction.

Excess iron aggravates the severity of COVID-19 infection.

Coronavirus disease-19 (COVID-19) can induce severe inflammation of the lungs and respiratory system. Severe COVID-19 is frequently associated with hyper inflammation and hyper-ferritinemia. High iron levels are known to trigger pro-inflammatory effects. Cumulative iron loading negatively impacts on a patients innate immune effector functions and increases the risk for infection related complications. Prognosis of severe acute respiratory SARS-CoV-2 patients may be impacted by iron excess. Iron is an essential co-factor for numerous essential cellular enzymes and vital cellular operations. Viruses hijack cells in order to replicate, and efficient replication requires an iron-replete host. Utilizing iron loaded cells in culture we evaluated their susceptibility to infection by pseudovirus expressing the SARS-CoV-2 spike protein and resultant cellular inflammatory response. We observed that, high levels of iron enhanced host cell ACE2 receptor expression contributing to higher infectivity of pseudovirus. In vitro Cellular iron overload also synergistically enhanced the levels of; reactive oxygen species, reactive nitrogen species, pro-inflammatory cytokines (IL-1ß, IL-6, IL-8 & TNF-α) and chemokine (CXCL-1&CCL-4) production in response to inflammatory stimulation of cells with spike protein. These results were confirmed using an in vivo mouse model. In future, limiting iron levels may be a promising adjuvant strategy in treating viral infection.

Intranasal cerium oxide nanoparticles improves locomotor activity and reduces oxidative stress and neuroinflammation in haloperidol-induced parkinsonism in rats.

Introduction: Cerium oxide nanoparticles (CONPs) have been investigated for their therapeutic potential in Parkinson's disease (PD) due to their potent and regenerative antioxidant activity. In the present study, CONPs were used to ameliorate the oxidative stress caused by free radicals in haloperidol-induced PD in rats following intranasal administration. Method: The antioxidant potential of the CONPs was evaluated in vitro using ferric reducing antioxidant power (FRAP) assay. The penetration and local toxicity of the CONPs was evaluated ex-vivo using goat nasal mucosa. The acute local toxicity of intranasal CONPs was also studied in rat. Gamma scintigraphy was used to assess the targeted brain delivery of CONPs. Acute toxicity studies were performed in rats to demonstrate safety of intranasal CONPs. Further, open field test, pole test, biochemical estimations and brain histopathology was performed to evaluate efficacy of intranasal CONPs in haloperidol-induced PD rat model. Results: The FRAP assay revealed highest antioxidant activity of prepared CONPs at a concentration of 25 µg/mL. Confocal microscopy showed deep and homogenous distribution of CONPs in the goat nasal mucus layers. No signs of irritation or injury were seen in goat nasal membrane when treated with optimized CONPs. Scintigraphy studies in rats showed targeted brain delivery of intranasal CONPs and acute toxicity study demonstrated safety. The results of open field and pole test showed highly significant (p < 0.001) improvement in locomotor activity of rats treated with intranasal CONPs compared to untreated rats. Further, brain histopathology of treatment group rats showed reduced neurodegeneration with presence of more live cells. The amount of thiobarbituric acid reactive substances (TBARS) was reduced significantly, whereas the levels of catalase (CAT), superoxide dismutase (SOD), and GSH were increased significantly, while amounts of interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-α) showed significant reduction after intranasal administration of CONPs. Also, the intranasal CONPs, significantly high (p < 0.001) dopamine concentration (13.93 ± 0.85 ng/mg protein) as compared to haloperidol-induced control rats (5.76 ± 0.70 ng/mg protein). Conclusion: The overall results concluded that the intranasal CONPs could be safe and effective therapeutics for the management of PD.

Advanced multifunctional nano-lipid carrier loaded gel for targeted delivery of 5-flurouracil and cannabidiol against non-melanoma skin cancer.

Non-melanoma skin cancer is one of the most common malignancies reported around the globe. Current treatment therapies fail to meet the desired therapeutic efficacy due to high degree of drug resistance. Thus, there is prominent demand in advancing the current conventional therapy to achieve desired therapeutic efficacy. To break the bottleneck, nanoparticles have been used as next generation vehicles that facilitate the efficient interaction with the cancer cells. Here, we developed combined therapy of 5-fluorouracil (5-FU) and cannabidiol (CBD)-loaded nanostructured lipid carrier gel (FU-CBD-NLCs gel). The current investigation has been designed to evaluate the safety and efficacy of developed 5-Flurouracil and cannabidiol loaded combinatorial lipid-based nanocarrier (FU-CBD NLCs) gel for the effective treatment of skin cancer. Initially, confocal microscopy study results showed excellent uptake and deposition at epidermal and the dermal layer. Irritation studies performed by IR camera and HET cam shows FU-CBD NLCs was much more tolerated and less irritant compared to conventional treatment. Furthermore, gamma scintigraphy evaluation shows the skin retention behavior of the formulation. Later, in-ovo tumor remission studies were performed, and it was found that prepared FU-CBD NLCs was able to reduce tumor volume significantly compared to conventional formulation. Thus, obtained results disclosed that permeation and disposition of 5-FU and CBD into different layers of the skin FU-CBD NLCs gel could be more potential carrier than conventional gel. Furthermore, prepared formulation showed greater tumor remission, better survival rate, reduction in tumor number, area, and volume with improved biochemical profile. Thus, prepared gel could serve as a promising formulation approach for the skin cancer treatment.

Molecular characterization of the unusual peptide CORO1C-47aa encoded by the circular RNA and docking simulations with its binding partner.

Circular RNAs, which have covalently closed ends, are in the class of non-coding RNAs. Recent studies reveal that they are associated with various biochemical pathways. One such involvement of circular RNAs is in the onset of different types of cancers. Though the circular RNAs are known as non-coding RNAs, some of them are found to possess the capacities to code for proteins. One such circular RNA is hsa-circ-0000437 which is known to code for a short peptide referred to as CORO1C-47aa. The peptide has anti-angiogenic activity and is associated with the prevention of endometrial cancer. The peptide binds to the PAS-B domain of the Aryl hydrocarbon Receptor Nuclear Translocator (ARNT). However, till date only the amino acid sequence of the peptide is known and no structural details of the peptide are available. Therefore, in this work, our aim was to predict how the peptide would fold and what could be its possible ligand binding sites. We used computational tools to determine the structure of the peptide refined further by molecular dynamics simulations. We then performed molecular docking simulations of the peptide with its known binding partner ARNT to gain an insight into the modes of binding as the process is associated with endometrial cancer. The possible ligand binding sites along-with the natures of the possible other different ligands of the peptide were analyzed further. From this structure function analysis study, we tried to elucidate the plausible mechanism of the involvements of the peptide in the onset of endometrial cancer. This is the first report on the structural characterization of the peptide and its modes of interactions with the partner protein ARNT. This study may therefore be useful in determining the structures of new drug candidates for the treatment of endometrial cancer.