CYP2C19*17 association with higher plasma 4-hydroxy tamoxifen in Pakistani (estrogen-positive) breast cancer patients.

Breast cancer (BC) continues to be the most common cancer in the women worldwide. Since estrogen receptor (ER)-positive BC accounts for the majority of newly diagnosed cases, endocrine therapy is advised to utilize either tamoxifen (Tam) or aromatase inhibitors. The use of Tam as a monotherapy or in conjunction with an aromatase inhibitor following two or three years of endocrine therapy has long been recommended. When used adjuvantly, Tam medication reduces BC mortality and relapses, while it extends survival times in metastatic BC when used in conjunction with other treatments. Unfortunately, the efficiency of Tam varies considerably. This study was conducted to explore the influence of genetic polymorphisms in CYP2C19 gene on Tam's pharmacogenetics and pharmacokinetics in estrogen-positive BC patients. Data from healthy, unrelated individuals (n = 410; control group) and ER-positive BC patients (n = 430) receiving 20 mg of Tam per day were recruited. Steady-state plasma concentrations of Tam and its three metabolites were quantified using the high-performance liquid chromatography in the patients. The CYP2C19 polymorphisms were genotyped using an Amplification Refractory Mutation System-Polymerase Chain Reaction (ARMS-PCR) approach. More than 65% of healthy individuals were extensive metabolizers (*1/*1) for CYP2C19, whereas more than 70% of ER-positive BC patients were rapid and ultrarapid metabolizers (*1/17*, *17/17*). The polymorphism CYP2C19*17 is significantly associated with higher 4-hydroxytamoxifen (4-OH-Tam). Patients with the *17/*17 genotype exhibited 1- to 1.5-fold higher 4-OH-Tam, which was also high in patients with the *1/*2 and *2/*2 genotypes.

Microbial production of docosahexaenoic acid (DHA): biosynthetic pathways, physical parameter optimization, and health benefits.

Omega-3 fatty acids, including docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and α-linolenic acid (ALA), are essential polyunsaturated fatty acids with diverse health benefits. The limited conversion of dietary DHA necessitates its consumption as food supplements. Omega-3 fatty acids possess anti-arrhythmic and anti-inflammatory capabilities, contributing to cardiovascular health. Additionally, DHA consumption is linked to improved vision, brain, and memory development. Furthermore, omega-3 fatty acids offer protection against various health conditions, such as celiac disease, Alzheimer's, hypertension, thrombosis, heart diseases, depression, diabetes, and certain cancers. Fish oil from pelagic cold-water fish remains the primary source of omega-3 fatty acids, but the global population burden creates a demand-supply gap. Thus, researchers have explored alternative sources, including microbial systems, for omega-3 production. Microbial sources, particularly oleaginous actinomycetes, microalgae like Nannochloropsis and among microbial systems, Thraustochytrids stand out as they can store up to 50% of their dry weight in lipids. The microbial production of omega-3 fatty acids is a potential solution to meet the global demand, as these microorganisms can utilize various carbon sources, including organic waste. The biosynthesis of omega-3 fatty acids involves both aerobic and anaerobic pathways, with bacterial polyketide and PKS-like PUFA synthase as essential enzymatic complexes. Optimization of physicochemical parameters, such as carbon and nitrogen sources, pH, temperature, and salinity, plays a crucial role in maximizing DHA production in microbial systems. Overall, microbial sources hold significant promise in meeting the global demand for omega-3 fatty acids, offering an efficient and sustainable solution for enhancing human health.

Induction Chemotherapy Response in Childhood Acute Lymphoblastic Leukaemia and its Correlation with Cytogenetic and Molecular Features.

OBJECTIVE: To study the correlation of cytogenetic and molecular abnormalities on induction chemotherapy in childhood acute lymphoblastic leukaemia (ALL). STUDY DESIGN: Analytical study. PLACE AND DURATION OF STUDY: Department of Haematology, Armed Forces Institute of Pathology (AFIP), from March 2021 to August 2021. METHODOLOGY: Patients aged 1-18 years with newly diagnosed acute lymphoblastic leukaemia were inducted. Patients aged less than 1 year and more than 18 years were excluded from the study. The diagnosis was based on morphology, cytochemistry, flow cytometry, and cytogenetic/molecular analysis. Risk stratification was done on the basis of age, TLC, and cytogenetic/molecular defects. The UKALL 2011 protocol was used for treatment with regimen-A for standard risk and regimen-B for high-risk patients. Bone marrow was repeated on day 29 of induction therapy and blast percentage was assessed to establish post-induction remission. Association between cytogenetic / molecular abnormalities and post-induction remission status was analysed using chi-square test. RESULTS: There were total 142 patients with mean age of 6.4 + 3.6 years and a male- to-female ratio of 2.7:1. Immunophenotyping revealed 85.9% cases as B-cell ALL and 14.1% as T-cell ALL. The most frequent cytogenetic and molecular abnormalities were hyperdiploidy (19%), t(9;22)/BCR-ABL1(p190) (10.6%), complex karyotype (5.6%), E2A-PBX1 (8.5%), and TEL-AML1 (4.9%). A total of 127/142 (89.4%) achieved haematological remission after induction therapy with two deaths during induction therapy (1.4%). Post-induction remission rate in patients with favorable cytogenetic/molecular defects was 100% and in children with bad prognostic changes, the rate of remission was 69.2%. Chi-square test showed a significant association between cytogenetic/molecular abnormalities and post-induction remission (p-value <0.001). CONCLUSION: Cytogenetic and molecular abnormalities have a significant association with post-induction remission in children with acute lymphoblastic leukaemia. KEY WORDS: Acute lymphoblastic leukaemia, Cytogenetics, Chemotherapy, Induction, Remission.

Laboratory Evaluation and Pathological Features of Bone Marrow Metastasis in Non-haematological Malignancies.

This study aimed to analyse the diagnostic accuracy of different laboratory parameters that can predict bone marrow metastasis. A cross-sectional analytical study was conducted at the Armed Forces Institute of Pathology (AFIP), Rawalpindi from March 2021 to August 2021. Bone marrow aspirates and biopsy procedures were done on 60 newly diagnosed cases of non-haematological malignancies as part of staging. Laboratory parameters noted for the study included peripheral blood smear findings, serum lactate dehydrogenase (LDH), radiological findings, and bone marrow aspirate/trephine biopsy results. Bone marrow metastasis was seen in 21/60 patients. The most common malignancies with bone marrow involvement were retinoblastoma and neuroblastoma. Laboratory findings showed no significant statistical difference in mean haemoglobin and total leukocyte count between cases and controls. Positive cases had a mean platelet count of 261.7 x 109/L and mean LDH of 750.1 U/L (p <0.05) for both parameters. ROC analysis showed the area under the curve (AUC) for LDH to be 0.969 (highly significant) showing a strong predictive value of LDH. Positive radiological findings were detected in only one case with bone marrow metastasis. The elevated level of serum LDH is not only cost-effective but also has high diagnostic accuracy to predict bone marrow metastasis. Key Words: Bone marrow, Biochemical, Lactate dehydrogenase, Metastasis, Non-haematological malignancies.

Biochemical characterization, cytotoxic, antimutagenic, anticancer and molecular docking studies on Tecomella undulata.

In this study bioassay-guided screening of Tecomella undulate was performed for its cytotoxic, antimutagenic and anticancer potential. The ariel parts were extracted on a polarity basis (methanol, dichloromethane and hexane). The in vivo toxicity was assessed on Caenorhabditis elegans, and its locomotion was affected by Tecomella undulata hexane (TUAH) the most. Ames test for antimutagenicity showed Tecomella undulata methanol (TUAM) exhibited against mutagen 2AA showed inhibition of 71.03% and 26.32% 2AA in TA98 while in in vitro MTT assay on carcinoma cell lines TUAM showed 68.1% cytotoxicity. Moreover, In resazurin assay on fibroblast cells African green monkey kidney VERO and on the panel of carcinoma cell lines, the most effective extract was TUAM on liver HepG-2 with CC50 value 117.37 ± 4.73 µg/ml followed by on lungs A549 with 142.01 ± 5.3. Furthermore, for the bioassay-guided screening, the selectivity index was calculated for TUAM CC50 ratio on HepG-2 and VERO which showed a decent 2.77 score. After column chromatography, the fraction TU-63 should remarkable cytotoxic effect in dose-response manner assay as (Hep-G2) CC50 value 11. 67 ± 1.37 µg/ml followed by (A549) CC50 value 17.23 ± 0.58 µg/ml. For qualitative analysis of anticancer potential LC-ESI-MS/MS the potential phytochemicals were identified. In silico molecular modelling against selected carcinogenic proteins. The results suggest Tecomella undulate the substantial anticancer potential which supports potential natural anticancer therapeutic drug candidate development for combating cancer.

In silico bimolecular characterization of anticancer phytochemicals from Fagonia indica.

The in silico molecular dynamics and structure-based site-specific drug design of indigenous plant biomolecules and selected proteins have remarkable potential for cancer therapy. A set of five proteins included for this research were epidermal growth factor protein (PDB ID; 1M17), crystal structure of mutated EGFR kinase (PDB ID; 2EB3), crystal structure of Bcl-xl (PDB ID; 2YXJ), apoptosis regulator protein MCL-1 BH3 (PDB ID; 3MK8) and apoptosis proteins (PDB ID; 5C3H). The present study on in silico investigation of fifteen indigenous medicinal plants were selected there one hundred thirty four ligands available literature were docked against five proteins involved in carcinogenesis. The highest scoring in silico plant, Fagonia indica was subjected to in vitro cytotoxic effects on HCT116, HepG-2 and HeLa human carcinoma cell lines. Molecular dynamics showed best ligand-protein inhibition interaction between Coumarin-2xyj and Kaempferol-2eb3 with promising binding affinities. Whereas, on HeLa human cervical cancer cell line IC50 was 28.3±0.102/ml. Fagonia indica could be potential source from natural products that have cytotoxic properties against cervical cancer cells by blocking mutant epidermal growth factor tyrosine or peroxisome proliferators activated receptor proteins.

In vitro antimutagenic, cytotoxic and anticancer potential of Fagonia indica phytochemicals.

Cancer is one of the most diagnosed and life threatening disease throughout the world. Nevertheless present day clinical management for cancers are surgery, radiations which are insufficient to contain the disease burden. In the past two decades, more than half of chemotherapeutic drugs developed are either directly or indirectly dependent on medicinal base phytocompounds or their derivative. The present study aims to provide the base for chemotherapeutic phytochemicals. Fagonia indica showed significant antimutagenic potential with reference to control IC50 values were calculated as 146.33±5.2µg/ml, TA100 (AZS) 105.33±4.0µg/ml, TA98 (2AA) 113.6±5.2µg/ml followed and TA98 (AZS) 112.6±4.4 in Ames test. For this reason, the antiproliferation effect of extracts on cancer cell lines was studied through resazurin fluorescence. On HepG-2 cell lines 50% cytotoxic concentration (CC50) of (FIWM) was recorded as 128.3±,2.43µg/ml. On the homo sapiens epithelial cell of lung tissue (A549), the high throughput instrumental analysis of Fagonia indica depicts maximum cytotoxic effect in 30hr. The electrical impedance displays the real-time evidence about qualitative apoptosis expressed. The impedance results were supported as palmitic acid from Fagonia indica virtually that inhibits Cyclin Dependent Kinase 2 (CDKs 2) in silico molecular docking studies. Fagonia indica extract possesses substantial antimutagenic, cytotoxic and anticancer activity which supports the potential of its phytochemicals for drug development.

Myxedema ascites complicated by ischemic colitis.

Myxedema Ascites is a rare finding of primary hypothyroidism, thereby leading to delayed diagnosis. However, prompt treatment with levothyroxine leads to complete resolution of the condition. We present a rare case of myxedema ascites in an elderly female and highlight the importance of early diagnosis and management. We also present ischemic colitis in the same patient, which has not been reported thus far in literature as a complication of myxedema ascites.

Patient Satisfaction and Understanding of Moderate Sedation During Endoscopy.

Millions of endoscopic procedures are performed in the US every year and the use of procedural sedation analgesia (PSA) is increasing with more procedures being performed outside the operating theater and gaining popularity due to reduced costs. Patients having endoscopic procedures usually expect that they would be deeply sedated during the procedure despite verbal counseling during pre-procedure clinic visits and are often dissatisfied with procedural awareness and discomfort. In order to better educate patients, written supplementary reading material was provided to the patients, which stated a clear goal of comfort during the procedure rather than deep sedation. The results showed that the written supplementary material did not improve the patient's understanding or remembrance of being counseled about moderate sedation. We emphasize that there is no substitute for a physician's repetitive verbal counseling.

Outcomes of Upper Gastrointestinal Bleeding Based on Time to Endoscopy: A Retrospective Study.

Introduction Non-variceal upper gastrointestinal bleeding (UGIB) is a major burden on the health care system. The timing of endoscopy has been an ongoing debate and data on the association of early endoscopy with a better or worse clinical outcome are conflicting. In our study, we aimed to identify the benefits versus the risks of performing an urgent endoscopy in regards to the number of endoscopic interventions, length of hospital stay, number of packed red blood cells (PRBCs) transfused, and mortality.  Methodology This is a retrospective record-based study. A total of 806 charts were reviewed and 251 patients with the signs and symptoms of UGIB on presentation were included in the study. Patients with variceal bleeding, lower gastrointestinal bleeding, insignificant bleeds with no drop in H/H, GI bleed not being the presenting complaint on admission, and patients on anticoagulation were excluded. Results Out of the patients who underwent an urgent esophagogastroduodenoscopy (EGD), 26.2% needed a second-look EGD 48 hours after the first EGD when compared to 4% and 2% in the early (12-24 hours) and late (>24 hours) endoscopy groups, respectively. In patients who underwent urgent EGD, 23% had active bleeding and it was statistically significant when compared to the other groups. The active bleeding limited the visualization during the endoscopy, which led to a repeat EGD in the urgent EGD group. If an endoscopic intervention was received, patients having EGD >24 hours received a smaller number of interventions. There was no statistical difference in the Blatchford scores between the three groups, indicating that the groups were similar in morbidity. No difference in mortality, hospital length of stay, or number of blood transfusions received, surgical or interventional radiology-guided interventions was found between the three groups. Conclusion Patients who underwent urgent endoscopy had more procedures, with no difference in mortality, number of units of blood transfused, or length of hospitalization when compared to the early or late endoscopy groups.

Virtual screening of antitumour phytochemical against peroxisome proliferators activated receptor proteins PPARs.

Cancers are caused by the defects in apoptosis process which leads to uncontrolled proliferation, therefore, most attractive drug target discovery strategy is to find ligands which have the ability to activate or regulate the apoptotic machinery. Peroxisome-proliferator-activated receptors (PPARs) are nuclear hormone receptors their over expression is observed in many tumours and contributes to chemotherapy resistance. The goal of this study to scrutinized antitumor phytochemicals from Alysicarpus bupleurifolius, Piper nigrum and Plumeria obtuse and potential energy values render from interactions between active site residues and ligands. The potential phytochemicals with significant binding affinity are ursolic acid, cis-4-decenoic acid and p-coumaric acid respectively most effective compounds in high throughput virtual screening belongs to Plumeria obtuse against PPARs associated with tumour development and progression. This modern drug designing modeling in silico approach, therefore, identifies the potential leads against over expressed tumours.

Pembrolizumab-Induced Mobitz Type 2 Second-Degree Atrioventricular Block.

Pembrolizumab is a monoclonal antibody directed towards programmed cell death protein 1 (PD-1) and is an antineoplastic drug which has a growing variety of oncologic uses. Pembrolizumab is commonly associated with immune-related adverse events (IRAEs) but is infrequently noted to cause cardiotoxicities such as myocarditis, arrhythmias, and heart failure. The following case report illustrates the clinical course of a 67-year-old female patient with stage IV non-small-cell lung cancer who developed Mobitz type 2 second-degree atrioventricular block three weeks after receiving her first infusion of pembrolizumab. Within a few hours of presentation, she progressed to symptomatic complete heart block requiring emergent placement of a temporary transvenous pacemaker. The article further discusses proposed mechanisms to explain IRAEs and management of IRAEs. We conclude by recommending a higher degree of caution and awareness among all physicians when treating patients on immunotherapy and a multidisciplinary approach when considering resumption of immune checkpoint inhibitor therapy.

In-vitro antiviral activity of Solanum nigrum against Hepatitis C Virus.

BACKGROUND: Hepatitis C is a major health problem causes liver cirrhosis, hepatocellular carcinoma and death. The current treatment of standard interferon in combination with ribavirin, has limited benefits due to emergence of resistant mutations during long-term treatment, adverse side effects and high cost. Hence, there is a need for the development of more effective, less toxic antiviral agents. RESULTS: The present study was designed to search anti-HCV plants from different areas of Pakistan. Ten medicinal plants were collected and tested for anti-HCV activity by infecting the liver cells with HCV 3a innoculum. Methanol and chloroform extracts of Solanum nigrum (SN) seeds exhibited 37% and more than 50% inhibition of HCV respectively at non toxic concentration. Moreover, antiviral effect of SN seeds extract was also analyzed against HCV NS3 protease by transfecting HCV NS3 protease plasmid into liver cells. The results demonstrated that chloroform extract of SN decreased the expression or function of HCV NS3 protease in a dose- dependent manner and GAPDH remained constant. CONCLUSION: These results suggest that SN extract contains potential antiviral agents against HCV and combination of SN extract with interferon will be better option to treat chronic HCV.