Despite the growth of molecular diagnosis from the era of Hippocrates, the emergence of COVID-19 is still remarkable. The previously used molecular techniques were not rapid enough to screen a vast population at home, in offices, and in hospitals. Additionally, these techniques were only available in advanced clinical laboratories.The pandemic outbreak enhanced the urgency of researchers and research and development companies to invent more rapid, robust, and portable devices and instruments to screen a vast community in a cost-effective and short time. There has been noteworthy progress in molecular diagnosing tools before and after the pandemic. This review focuses on the advancements in molecular diagnostic techniques before and after the emergence of COVID-19 and how the pandemic accelerated the implantation of molecular diagnostic techniques in most clinical laboratories towardbecoming routine tests.

Introduction: The receptor activator NF-κB ligand (RANKL) and Osteoprotegrin (OPG) single nucleotide polymorphisms (SNPs) have been associated with the risk of breast cancer to bone metastasis. This study was designed to investigate the association of RANKL and OPG gene polymorphisms with breast to bone metastasis in Pashtun population of Khyber Pakhtunkhwa, Pakistan. Materials and methods: A total of 215 participants were enrolled containing 106 breast cancer patients, 58 breast to bone metastasis and 51 age and gender matched healthy controls. RANKL (rs9533156) and OPG (rs2073618, rs3102735) polymorphisms were genotyped in genomic DNA, using Tetra-ARMS PCR protocol. The results were analyzed among the three groups and P-value less then 0.05 were considered statistically significant. Results: Our results displayed significant association of OPG (rs3102735) risk allele and corresponding genotypes in breast cancer vs healthy controls, bone metastasis vs healthy controls and breast cancer vs breast to bone metastasis as a disease risk. However, there was no association observed for OPG (rs2073618) risk allele and corresponding genotypes with the diseases risk. Similarly, RANKL (rs9533156) risk allele and corresponding genotypes in breast cancer vs healthy controls, bone metastasis vs healthy controls and breast cancer vs breast to bone metastasis exhibited significant association except for the risk allele carrying genotypes in breast to bone metastasis. Conclusion: OPG (rs3102735) and RANKL (rs9533156) exhibited significant association with breast to bone metastasis while OPG (rs2073618) didn’t show significant association with breast to bone metastasis in Pashtun population of Pakistan. However, this study unlocks more questions to investigate the exact scenario of genetic predisposition of breast to bone metastasis.

Background: Plant-derived products or extracts are widely used in folk/traditional medicine to treat several infections, ailments, or disorders. A well-known medicinal herb, Myrtus communis is an evergreen fragrant plant native to the Mediterranean region that has been used for ages in traditional medicine around the world. Materials and methods: The microplate alamarBlue assay and the well diffusion method were used to evaluate the zone of inhibition and MIC, respectively. The double-disc diffusion method was used to investigate the synergy between antibiotics and the extract. The crystal violet method was used to investigate biofilm development. The SulphoRhodamine-B assay and DNA flow cytometry were used to investigate the proliferation and subsequent distribution of cells among different phases of the cell cycle. The apoptotic and necrotic phases of the cancer cells were examined using flow cytometry in conjunction with Annexin V-FITC/PI labeling. Using the IBM SPSS statistical program, a one-way ANOVA with Tukey’s post hoc test was employed for statistical analysis. Results: The ethanolic leaf extract of M. communis showed a strong growth inhibition effect (zone of inhibition: 20.3 ± 1.1–26.3 ± 2.5 mm, MIC: 4.88–312.5 µg/mL, and MBC: 39.07–1250 μg/mL) against several rapidly growing and slow-growing mycobacterial strains in a dose-dependent manner. Damage to the cell wall of bacterial cells was determined to be the cause of the antimycobacterial action. The extract inhibited biofilm formation (MBIC of 9.7 µg/mL) and eradicated already-formed mature and ultra-mature biofilms of M. smegmatis, with MBEC values of 78 µg/mL and 156 µg/mL, respectively. Additionally, the extract exhibited potent anticancer effects against diverse cancer cell lines of the breast (MCF-7), liver (HepG2), cervix (HeLa), and colon (HCT116) (IC₅₀ for HCT116: 83 ± 2.5, HepG2: 53.3 ± 0.6, MCF-7: 41.5 ± 0.6, and HeLa: 33.3 ± 3.6) by apoptosis after arresting the cells in the G₁ phase of the cell cycle. Conclusions: These results suggest that M. communis leaf extract is a potential source of secondary metabolites that could be further developed as potential anticancer and antimycobacterial agents to treat diverse types of cancers and mycobacterial infections.

Skin cancer is a growing global health concern, marked by high incidence and significant mortality, particularly in aggressive melanoma subtypes. In this study, we employed an integrative network pharmacology and molecular docking approach to evaluate the anticancer potential of Nigella sativa (black seed) against skin cancer. Initially, 13 active compounds were identified from N. sativa based on stringent pharmacokinetic criteria. Target prediction using SwissTargetPrediction, integrated with 9697 skin cancer-associated genes from GeneCards and DisGeNET, revealed 303 overlapping targets implicated in critical oncogenic processes. Subsequent Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses via the DAVID database identified 780 biological processes, 87 cellular components, and 278 molecular functions. Enriched pathways included the positive regulation of the MAPK cascade, EGFR signaling, angiogenesis, and several other pathways central to skin cancer pathogenesis. The compound-target network further underscored the polypharmacological nature of N. sativa, highlighting hub genes. Molecular docking studies were conducted to validate the interactions of select bioactive compounds with key receptors (AR, CDK4, EGFR, MAPK1, and MAPK3). Among the compounds, Gramisterol (CID: 5283640) demonstrated the strongest binding affinities, with energies of − 9.1 kcal/mol for both EGFR and MAPK3, − 9.0 kcal/mol for CDK4, − 8.2 kcal/mol for MAPK1, and − 7.4 kcal/mol for AR. Cycloeucalenol (CID: 101690), Obtusifoliol (CID: 65225) and Lophenol (CID: 160482) also exhibited potent interactions, particularly with EGFR, MAPK1, MAPK3 and CDK4, supporting their potential to disrupt tumor proliferation and survival signaling. Collectively, these findings indicate that N. sativa’s bioactive compounds can modulate multiple cancer-related pathways, offering a promising multi-target strategy for skin cancer therapy. This computational study lays a robust foundation for subsequent in vitro and in vivo validations and paves the way for the development of novel, less toxic therapeutic regimens against skin cancer. Although the computational discoveries offer a solid conceptual foundation, evidence of their clinical significance is still pending. The next crucial stage of this investigation is to conduct extensive in vitro and in vivo verification investigations in order to close this gap. These initiatives will be crucial to converting our research into practical skin cancer treatment plans.

Objective: The present investigation aims on the clinical attributes and haematological parameters between symptomatic (COVID– 19 ICU) and asymptomatic (COVID– 19 homes isolation) patients as predisposing sign for COVID-19 related mortality. Materials and Methods: A retrospective cohort research was conducted of admitted patients to ICU, who were suffering from severe COVID-19 in Aseer Central Hospital, Abha, Kingdom of Saudi Arabia (KSA) from July 2020 until September 2020. The study included individuals with COVID − 19 and ICU admission as symptomatic group and others who are COVID– 19 positives with quarantine as asymptomatic group. Epidemiological, clinical and haematological laboratory data were retrospectively collected, analysed with control subjects. Results: Of the 38 ICU patients studied, the most common symptoms were fever and respiratory distress (100%), cough (86.8%). Majority were of Saudi origin (78.9%). Eighteen (47.4%) COVID-19 ICU patients showed leukocytosis, 6 (15.8%) had severe thrombocytopenia (with most having thrombocytopenia), 18 (47.4%) were anaemic. A significant correlation was observed between the WBC, RBC, Hb, platelets, neutrophil and lymphocyte count between ICU inmates compared with quarantine (p < 0.001) and RBC, Hb, neutrophil and lymphocyte count with control groups (p < 0.001). Conclusion: From the observations it is evident that, the blood tests have potential clinical value in predicting COVID-19 progression. Further, patient characteristics including age, leukocyte count, RBC, platelets and differential leukocyte counts may be significant predictors for monitoring the progression of the critical illness observed in SARS-COV-2 patients. Also, treatment procedures can be re-defined further to reduce COVID– 19 mortalities in more critically ill COVID-19 individuals.