Knockdown of CSNK2β suppresses MDA-MB231 cell growth, induces apoptosis, inhibits migration and invasion

Breast cancer is the most common cancer among women worldwide. Among different types of breast cancer known, treatment of triple-negative breast cancer is a major challenge because of its aggressiveness and poor prognosis; thus, identification of specific drivers is required for targeted therapies of breast cancer malignancy. Protein Casein Kinase (CSNK) is a serine/threonine kinase that exists as a tetrameric complex consisting of two catalytic (α and /or α') and two regulatory β subunits. CSNK2β can also function independently without catalytic subunits and exist as a distinct population in cells. This study aims to elucidate the role of Casein Kinase 2β (CSNK2β) gene in cell proliferation, cell cycle, migration and apoptosis of triple-negative breast cancer MDA-MB-231 cells. The silencing of CSNK2β in MDA-MB-231 cells resulted in decreased cell viability and colony formation. Cell cycle analysis showed a significant arrest of cells in G2M phase. Hoechst and CM-H2DCFDA staining showed nuclear condensation and augmented intracellular reactive oxygen species (ROS) production. Furthermore, silencing of CSNK2β in MDA-MB-231 cells modulated the apoptotic machinery- BAX, Bcl-xL, and caspase 3; autophagy machinery-Beclin-1 and LC3-1; and inhibited the vital markers (p-ERK, c-Myc, NF-κB, E2F1, PCNA, p38-α) associated with cell proliferation and DNA replication pathways. In addition, knockdown of CSNK2β also affected the migration potential of MDA-MB-231, as observed in the wound healing and transwell migration assays. Altogether, the study suggests that CSNK2β silencing may offer future therapeutic target in triple-negative breast cancer

Aging-Associated miR-217 Aggravates Atherosclerosis and Promotes Cardiovascular Dysfunction.

microRNAs are master regulators of gene expression with essential roles in virtually all biological processes. miR-217 has been associated with aging and cellular senescence, but its role in vascular disease is not understood. Approach and Results: We have used an inducible endothelium-specific knock-in mouse model to address the role of miR-217 in vascular function and atherosclerosis. miR-217 reduced NO production and promoted endothelial dysfunction, increased blood pressure, and exacerbated atherosclerosis in proatherogenic apoE-/- mice. Moreover, increased endothelial miR-217 expression led to the development of coronary artery disease and altered left ventricular heart function, inducing diastolic and systolic dysfunction. Conversely, inhibition of endogenous vascular miR-217 in apoE-/- mice improved vascular contractility and diminished atherosclerosis. Transcriptome analysis revealed that miR-217 regulates an endothelial signaling hub and downregulates a network of eNOS (endothelial NO synthase) activators, including VEGF (vascular endothelial growth factor) and apelin receptor pathways, resulting in diminished eNOS expression. Further analysis revealed that human plasma miR-217 is a biomarker of vascular aging and cardiovascular risk. Our results highlight the therapeutic potential of miR-217 inhibitors in aging-related cardiovascular disease.V.G. de Yébenes was supported by Ramón y Cajal grant RYC-2009-04503 and AECC foundation grant INVES18013GARC and by the Universidad Complutense de Madrid. S.M. Mur and A.R. Ramiro are supported by Centro Nacional de Investigaciones Cardiovasculares (CNIC) funding. A.R. Ramiro was supported by the Spanish Ministerio de Ciencia e Innovación (PID2019-107551RB-I00), the Spanish Ministerio de Economía, Industria y Competitividad (SAF2013-42767-R and SAF2016-75511-R), and the European Research Council StG BCLYM. M. Salaices was supported by the Ministerio de Ciencia e Innovación (SAF2016-80305P) and with J. Miguel Redondo by Instituto de Salud Carlos III (CIBER de Enfermedades Cardiovasculares, CB16/11/00286 and CB16/11/00264) and Comunidad de Madrid (B2017/BMD-3676). V.G. de Yébenes was supported by Ministerio de Ciencia e Innovación (PID2019-107551RB-I00). Further support was provided by the European Social Fund and the European Regional Development Fund “A Way to Build Europe.” The CNIC is supported by Ministerio de Ciencia, Innovacion y Universidades, and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (SEV-2015-0505).S

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P < 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Targeting Mek1/2-Erk1/2 signaling pathway in pancreatic cancer

El cáncer de páncreas es una de las enfermedades más letales, siendo la cuarta causa de muerte por cáncer en países occidentales. Con una supervivencia global a los 5 años de alrededor del 5%, el tratamiento estándar actual consiste en quimioterapia, y la amplia mayoría de terapias dirigidas que se han probado hasta ahora no han tenido éxito. Por ello, es necesario el desarrollo de nuevas terapias contra componentes clave de esta enfermedad, así como que combatan los mecanismos de resistencia. Alrededor del 90% de los cánceres de páncreas contienen mutaciones en K-Ras, clave para el desarrollo de esta enfermedad. K-Ras es una GTPasa que transduce señales iniciadas en receptores tirosina quinasa (RTKs) y controla proliferación celular y supervivencia, entre otras funciones. A pesar del desarrollo de inhibidores específicos de K-Ras durante las últimas décadas, ninguno ha conseguido demostrar eficacia en la clínica. Como consecuencia, los esfuerzos se han dirigido a inhibir a las proteínas reguladas por K-Ras, como las Raf o las Mek. Sin embargo, a pesar de que los inhibidores de B-Raf y Mek1/2 han demostrado eficacia clínica en melanoma, en cáncer de páncreas no se ha observado tal eficacia. En esta tesis, probamos la eficacia de la inhibición de Mek1/2 en modelos de cáncer de páncreas, en combinación con quimioterapia, utilizando líneas celulares y xenoinjertos derivados de pacientes con cáncer páncreas (PC-PDXs). Sin embargo, demostramos que este efecto se ve limitado por la aparición de clones resistentes, que resultan como consecuencia del alto grado de heterogeneidad de estos tumores. En este trabajo también se muestra que, cuando las células de cáncer de páncreas adquieren resistencia a la inhibición de Mek1/2, aumentan la expresión del factor de transcripción Slug. Este factor de transcripción induce resistencia, con un mecanismo de acción que consiste en desacoplar la división celular del control de la ruta de señalización Raf-Mek-Erk. Además, la expresión de Slug correlaciona con resistencia en células de cáncer de páncreas y melanoma. Asimismo, también mostramos que Slug controla la capacidad metastásica de las células de resistentes y es un indicador de mal pronóstico en pacientes con cáncer de páncreas y melanoma.Pancreatic cancer is one of most lethal human diseases, being the fourth leading cause of cancer-related deaths in western countries. With a 5-year overall survival of around 5%, the development of novel therapies is needed. The current standard treatment for this disease consists of chemotherapy, and many targeted therapies have failed to improve patients’ survival so far. For this reason, the development of novel therapies targeting key components of this cancer as well as the study of resistance becomes necessary. 90% of pancreatic cancers are mutated in the GTPase K-Ras, required for the development of this disease. K-Ras is a GTPase that transduces signals from tyrosine kinase receptors (RTKs) and controls cell proliferation and survival. Despite the initial approaches for pharmacologically inhibiting K-Ras started more than twenty years ago, none of the inhibitors has succeed in the clinics so far. As K-Ras is still considered ‘undruggable’, the efforts have been focused on targeting its downstream effectors, as the Raf and Mek proteins. Despite B-Raf and Mek1/2 inhibitors have improved patients’ survival in melanoma, they have failed to succeed in pancreatic cancer patients. In this work, we assess the efficacy of Mek1/2 inhibition against pancreatic cancer. Using cell lines and pancreatic cancer-derived xenografts (PC-PDXs), we gauged the efficacy of the Mek1/2 inhibitor MEK162 in addition to the standard chemotherapy and we show that it impairs growth in vitro and in vivo. However, we also demonstrate that the effectiveness of Mek1/2 inhibition is limited by the emergence of resistant clonal populations, that result from the high degree of tumor heterogeneity and they compromise the effectiveness of this treatment. In this thesis, we also demonstrate that, when pancreatic cancer cells acquire resistance to Mek1/2 inhibition they increase the expression of the EMT zinc finger transcription factor Slug. This transcription factor regulates resistance to Mek1/2 inhibitors in pancreatic cancer cells by uncoupling the regulation of cell division by the Mek1/2-Erk1/2 pathway, and it correlates with resistance in a panel of pancreatic cancer and melanoma cell lines. Likewise, Slug is responsible for the enhanced metastatic ability of resistant cells, and it consistently correlates with poor survival in pancreatic cancer and melanoma patients

The Frequency of Common Complications of Veress Needle in Laparoscopic Cholecystectomy

METHODOLOGYThe study was conducted in department of surgery, Lady Reading Hospital Peshawar within six months and it was descriptive cross sectional study. In this study a total of 177 patients were observed. The age ranges were 21 to 65 years and male to female ratio was 1:2. The most common complication was abdominal wall hemorrhage in 22% patients while 18% patients had omental injury. The abdominal wall emphysema was observed in 10% patients.OBJECTIVETo determine the frequency of common complications of Veress Needle used for creating pneumoperitoneum in laparoscopic cholecystectomyCONCLUSIONOur study showed that veress needle technique is safe, easy and cost effective for primary access to create pneumoperitonium

Impact of Education on Poverty Reduction on Poverty in Afghanistan

This research aimed to explore the pivotal role of education in alleviating poverty in Afghanistan. Employing a quantitative research approach, the study utilized random sampling with a sample size of 302 individuals, employing Morgan's method. The data was collected through the employment of the Likert spectrum questionnaire, which demonstrated a commendable accuracy rate of 91% as determined by Cronbach's alpha test.to ensure rigorous analysis and precise inference, various statistical techniques were employed, including stepwise regression analysis, factor analysis, internal consistency, Kaiser-Meyer-Olkin, and Bartlett's test. The research findings revealed several significant and positive impacts of education on poverty reduction. These include the creation of job opportunities through knowledge, rapid technological adaptations, increased employment rates, enhanced access to financial resources, augmented household income, and improved access to production factors through knowledge acquisition. Moreover, the research findings firmly reject the null hypothesis and instead lend support to the alternative hypothesis, underscoring the substantial influence of education on poverty reduction. Consequently, it is highly recommended to prioritize education as a key strategy in combating poverty in Afghanistan. By doing so, the nation can empower individuals, foster economic growth, and create a brighter future for its citizens

Optimizing ChatGPT as a Writing Aid for EFL Learners: Balancing Assistance and Skill Development in Writing Proficiency

This study explores the strategic incorporation of advanced language generation models, like ChatGPT, in the context of EFL education to enhance writing proficiency. The research delves into the intricate relationship between the use of ChatGPT as a writing aid and the development of autonomous writing skills in EFL learners. The primary aim is to optimize ChatGPT's utilization, finding a delicate balance between leveraging its support for improving grammar, vocabulary, and composition, while also promoting learners' self-sufficiency in crafting logically structured and coherent written discourse. By conducting a thorough analysis of various pedagogical approaches, this research seeks to offer practical insights to educators regarding effective strategies for incorporating ChatGPT to improve the writing abilities of EFL learners, thus enriching the broader landscape of language education. The empirical findings of this study are intended to inform the pedagogical community and advance discussions on the effective assimilation of AI-powered writing aids in language education paradigms, ultimately guiding their optimal use in instructional settings

Targeting Mek1/2-Erk1/2 signaling pathway in pancreatic cancer

El cáncer de páncreas es una de las enfermedades más letales, siendo la cuarta causa de muerte por cáncer en países occidentales. Con una supervivencia global a los 5 años de alrededor del 5%, el tratamiento estándar actual consiste en quimioterapia, y la amplia mayoría de terapias dirigidas que se han probado hasta ahora no han tenido éxito. Por ello, es necesario el desarrollo de nuevas terapias contra componentes clave de esta enfermedad, así como que combatan los mecanismos de resistencia. Alrededor del 90% de los cánceres de páncreas contienen mutaciones en K-Ras, clave para el desarrollo de esta enfermedad. K-Ras es una GTPasa que transduce señales iniciadas en receptores tirosina quinasa (RTKs) y controla proliferación celular y supervivencia, entre otras funciones. A pesar del desarrollo de inhibidores específicos de K-Ras durante las últimas décadas, ninguno ha conseguido demostrar eficacia en la clínica. Como consecuencia, los esfuerzos se han dirigido a inhibir a las proteínas reguladas por K-Ras, como las Raf o las Mek. Sin embargo, a pesar de que los inhibidores de B-Raf y Mek1/2 han demostrado eficacia clínica en melanoma, en cáncer de páncreas no se ha observado tal eficacia. En esta tesis, probamos la eficacia de la inhibición de Mek1/2 en modelos de cáncer de páncreas, en combinación con quimioterapia, utilizando líneas celulares y xenoinjertos derivados de pacientes con cáncer páncreas (PC-PDXs). Sin embargo, demostramos que este efecto se ve limitado por la aparición de clones resistentes, que resultan como consecuencia del alto grado de heterogeneidad de estos tumores. En este trabajo también se muestra que, cuando las células de cáncer de páncreas adquieren resistencia a la inhibición de Mek1/2, aumentan la expresión del factor de transcripción Slug. Este factor de transcripción induce resistencia, con un mecanismo de acción que consiste en desacoplar la división celular del control de la ruta de señalización Raf-Mek-Erk. Además, la expresión de Slug correlaciona con resistencia en células de cáncer de páncreas y melanoma. Asimismo, también mostramos que Slug controla la capacidad metastásica de las células de resistentes y es un indicador de mal pronóstico en pacientes con cáncer de páncreas y melanoma.Pancreatic cancer is one of most lethal human diseases, being the fourth leading cause of cancer-related deaths in western countries. With a 5-year overall survival of around 5%, the development of novel therapies is needed. The current standard treatment for this disease consists of chemotherapy, and many targeted therapies have failed to improve patients' survival so far. For this reason, the development of novel therapies targeting key components of this cancer as well as the study of resistance becomes necessary. 90% of pancreatic cancers are mutated in the GTPase K-Ras, required for the development of this disease. K-Ras is a GTPase that transduces signals from tyrosine kinase receptors (RTKs) and controls cell proliferation and survival. Despite the initial approaches for pharmacologically inhibiting K-Ras started more than twenty years ago, none of the inhibitors has succeed in the clinics so far. As K-Ras is still considered 'undruggable', the efforts have been focused on targeting its downstream effectors, as the Raf and Mek proteins. Despite B-Raf and Mek1/2 inhibitors have improved patients' survival in melanoma, they have failed to succeed in pancreatic cancer patients. In this work, we assess the efficacy of Mek1/2 inhibition against pancreatic cancer. Using cell lines and pancreatic cancer-derived xenografts (PC-PDXs), we gauged the efficacy of the Mek1/2 inhibitor MEK162 in addition to the standard chemotherapy and we show that it impairs growth in vitro and in vivo. However, we also demonstrate that the effectiveness of Mek1/2 inhibition is limited by the emergence of resistant clonal populations, that result from the high degree of tumor heterogeneity and they compromise the effectiveness of this treatment. In this thesis, we also demonstrate that, when pancreatic cancer cells acquire resistance to Mek1/2 inhibition they increase the expression of the EMT zinc finger transcription factor Slug. This transcription factor regulates resistance to Mek1/2 inhibitors in pancreatic cancer cells by uncoupling the regulation of cell division by the Mek1/2-Erk1/2 pathway, and it correlates with resistance in a panel of pancreatic cancer and melanoma cell lines. Likewise, Slug is responsible for the enhanced metastatic ability of resistant cells, and it consistently correlates with poor survival in pancreatic cancer and melanoma patients

Single Layer Extra-Mucosal Interrupted Anastomoses; Revalidated

Objective:To evaluate the safety regarding anastomotic failure of single layer interrupted extra mucosal intestinal anastomosis in comparison with double layer intestinal anastomosisMethodology:This prospective comparative study was conducted in surgical A unit of Lady reading Hospital Peshawar from 1st June 2007 to 1st February 2008 (8 months).Patients were divided into two groups, each comprising 60 patients. First 60 consecutive patients were included in Group A, for single layer extra mucosal anastomosis while Group B included last 60 consecutive patients for double layer inverting anastomosis (continuous inner and interrupted outer Lambert sutures). All the cases were admitted through OPD and emergency. The safety of two techniques of anastomosis was analyzed by comparing the outcome in terms of complications.Results:In this study, anastomosis leakage occurred only in 4 (3.33%) patients, one (1.67%) in group A and three (5%) in group B with a P-Value 0.138. Mean age of patient in group A was 36.15 years (+/- 6.0 years) and in group B was 33.25 years (+/- 5.5 years).Conclusion:Single layer extra-mucosal anastomosis has least anastomotic leakage and other complication like wound infection, septicemia, and collection and burst abdomen than in patients with double layer investing anastomosis