Identification of Phytochemicals from Arabian Peninsula Medicinal Plants as Strong Binders to SARS-CoV-2 Proteases (3CLPro and PLPro) by Molecular Docking and Dynamic Simulation Studies.

We provide promising computational (in silico) data on phytochemicals (compounds 1-10) from Arabian Peninsula medicinal plants as strong binders, targeting 3-chymotrypsin-like protease (3CLPro) and papain-like proteases (PLPro) of SARS-CoV-2. Compounds 1-10 followed the Lipinski rules of five (RO5) and ADMET analysis, exhibiting drug-like characters. Non-covalent (reversible) docking of compounds 1-10 demonstrated their binding with the catalytic dyad (CYS145 and HIS41) of 3CLPro and catalytic triad (CYS111, HIS272, and ASP286) of PLPro. Moreover, the implementation of the covalent (irreversible) docking protocol revealed that only compounds 7, 8, and 9 possess covalent warheads, which allowed the formation of the covalent bond with the catalytic dyad (CYS145) in 3CLPro and the catalytic triad (CYS111) in PLPro. Root-mean-square deviation (RMSD), root-mean-square fluctuation (RMSF), and radius of gyration (Rg) analysis from molecular dynamic (MD) simulations revealed that complexation between ligands (compounds 7, 8, and 9) and 3CLPro and PLPro was stable, and there was less deviation of ligands. Overall, the in silico data on the inherent properties of the above phytochemicals unravel the fact that they can act as reversible inhibitors for 3CLPro and PLPro. Moreover, compounds 7, 8, and 9 also showed their novel properties to inhibit dual targets by irreversible inhibition, indicating their effectiveness for possibly developing future drugs against SARS-CoV-2. Nonetheless, to confirm the theoretical findings here, the effectiveness of the above compounds as inhibitors of 3CLPro and PLPro warrants future investigations using suitable in vitro and in vivo tests.

Covalent Inhibitors from Saudi Medicinal Plants Target RNA-Dependent RNA Polymerase (RdRp) of SARS-CoV-2.

COVID-19, a disease caused by SARS-CoV-2, has caused a huge loss of human life, and the number of deaths is still continuing. Despite the lack of repurposed drugs and vaccines, the search for potential small molecules to inhibit SARS-CoV-2 is in demand. Hence, we relied on the drug-like characters of ten phytochemicals (compounds 1-10) that were previously isolated and purified by our research team from Saudi medicinal plants. We computationally evaluated the inhibition of RNA-dependent RNA polymerase (RdRp) by compounds 1-10. Non-covalent (reversible) docking of compounds 1-10 with RdRp led to the formation of a hydrogen bond with template primer nucleotides (A and U) and key amino acid residues (ASP623, LYS545, ARG555, ASN691, SER682, and ARG553) in its active pocket. Covalent (irreversible) docking revealed that compounds 7, 8, and 9 exhibited their irreversible nature of binding with CYS813, a crucial amino acid in the palm domain of RdRP. Molecular dynamic (MD) simulation analysis by RMSD, RMSF, and Rg parameters affirmed that RdRP complexes with compounds 7, 8, and 9 were stable and showed less deviation. Our data provide novel information on compounds 7, 8, and 9 that demonstrated their non-nucleoside and irreversible interaction capabilities to inhibit RdRp and shed new scaffolds as antivirals against SARS-CoV-2.

A new species and a key to the genus Leiurus Ehrenberg, 1828 (Scorpiones, Buthidae) from Saudi Arabia.

A new species, Leiurushadb Al-Qahtni, Al-Salem, Alqahtani & Badry, sp. nov., is described and illustrated from the Majami al-Hadb Protected Area in the Riyadh Province of Saudi Arabia. The new species is compared with species of Leiurus distributed in Saudi Arabia, especially L.arabicus Lowe, Yagmur & Kovarík, 2014. The integrated results indicate that the population found in Majami al-Hadb represents a distinct species, which is described herein. Moreover, the molecular analysis is conducted on the mitochondrial gene 16S rRNA to compare L.hadb sp. nov. with samples of L.arabicus and L.haenggii from Saudi Arabia. The analysis revealed a genetic divergence ranging from 6.0 to 12%. The combination of molecular evidence and morphological characteristics provides adequate support for recognizing the Majami al-Hadb population as a distinct species. Additionally, an identification key for the genus Leiurus found in Saudi Arabia is also provided.

Does Omega-3 Fatty Acid Supplementation Have Favorable Effects on the Lipid Profile in Postmenopausal Women? A Systematic Review and Dose-response Meta-analysis of Randomized Controlled Trials.

PURPOSE: Menopause is associated with disturbances in the metabolism of lipids. Moreover, during the postmenopausal period, female subjects are more prone to develop dyslipidemia. Omega-3 fatty acids, which exert cardioprotective, anti-inflammatory, and lipid-lowering actions, are commonly recommended in postmenopausal women. However, their effect on serum lipids in this population remains unclear. This systematic review and meta-analysis of randomized controlled trials (RCTs) was conducted to clarify this research question. METHODS: We systematically searched the Web of Science, Scopus, PubMed/MEDLINE, and EMBASE databases from their inception until January 3, 2022. The DerSimonian and Laird random-effects model was used to combine effect sizes. FINDINGS: Omega-3 fatty acid supplementation resulted in a decrease in triglyceride concentrations (weighted mean difference [WMD], -17.8 mg/dL; 95% CI, -26 to -9.6; P < 0.001), particularly in the RCTs that lasted ≤16 weeks (WMD, -18.6 mg/dL), when the baseline triglyceride concentrations were ≥150 mg/dL (WMD, -22.8 mg/dL), in individuals with a body mass index ≥30 kg/m2 (WMD, -19.3 mg/dL), and when the dose of omega-3 fatty acids was ≥1 g/d (WMD, -21.10 mg/dL). LDL-C (WMD, 4.1 mg/dL; 95% CI, 1.80 to 6.36; P < 0.001) and HDL-C (WMD, 2.1 mg/dL; 95% CI, 0.97 to 3.2; P < 0.001) values increased. Total cholesterol levels (WMD, -0.15 mg/dL; 95% CI, -4 to 3.74; P = 0.94) remained unchanged after administration of omega-3 fatty acids. IMPLICATIONS: In postmenopausal women, supplementation with omega-3 fatty acids resulted in a significant reduction in triglyceride concentrations and a modest elevation in HDL-C and LDL-C levels, whereas this intervention did not affect total cholesterol values.

Vitamin D protects against depression: Evidence from an umbrella meta-analysis on interventional and observational meta-analyses.

Meta-analyses of interventional and observational studies investigating the efficacy and the relationship between vitamin D and depression provided inconsistent results. The current umbrella meta-analysis was conducted to assess the available evidence and provide a conclusive outcome in this regard. The following international databases were systematically searched till March 2022: PubMed, Scopus, Embase, Web of Science, and Google Scholar. Random-effects model was carried out to calculate the pooled point estimates and their respective 95 % confidence intervals (CI). Ten meta-analyses of randomised controlled trials (RCTs) revealed significant reduction in depression symptoms comparing participants on vitmain D supplements to those on placebo (Pooled standardised mean difference: - 0.40; 95 % CI: - 0.60, - 0.21, p < 0.01: I2 = 89.1 %, p < 0.01). Four meta-analyses of cohort studies (with one having two subgroups) revealed that participants with lower levels of serum vitamin D were at increased odds of depression than those with higher levels of serum vitamin D (Pooled odds ratio: 1.60; 95 % CI: 1.08, 2.36, p < 0.01; I2 = 91.3 %, p < 0.01). The present umbrella meta-analysis confirms the potential benefits of vitamin D supplementation and higher serum vitamin D levels in reducing the development and symptoms of depression.

Contract Cheating and Ghostwriting among University Students in Health Specialties.

Contract cheating and ghostwriting are forms of misconduct that are unethical and a serious academic issue, especially among healthcare professionals, as they directly impact patient health. To date, research on this area in the Middle East has been limited. Therefore, we used a validated self-administered questionnaire to investigate the awareness, perceptions, and reasons for these behaviors among 682 students in health specialties at five universities in Riyadh, Saudi Arabia. The majority of the students (60.1%) were unaware of the terms "contract cheating" and "ghostwriting," and 69.5% had not received any prior training on integrity. However, having prior training had a positive effect on awareness levels, and respondents attending private universities were significantly more aware than those attending public universities. The factors that contributed to contract cheating behavior included poor time management, English language difficulties, and a lack of writing skills. These findings emphasize the need for integrity training at the national level to raise awareness.

Isosorbide mononitrate for cervical ripening during labour induction: A systematic review and meta-analysis of 23 randomized controlled trials.

OBJECTIVE: To conduct a systematic review and meta-analysis of all randomized controlled trials (RCTs) that evaluated the efficacy and safety of isosorbide mononitrate (IMN) in promoting cervical ripening during labour induction. METHODS: Six major databases were searched from inception until 22 April 2021. The risk of bias of included studies was assessed. Various endpoints (n = 21) were meta-analysed, and the endpoints were pooled as mean differences (MD) or risk ratios (RR) with 95% confidence intervals (CI). RESULTS: In total, 23 RCTs were included in this review, comprising 26 intervention arms and a total of 4305 patients (2210 and 2095 patients were allocated to the IMN and control groups, respectively). Pertaining to obstetric-related maternal outcomes, the pooled analysis showed that admission to delivery time and rate of caesarean delivery were significantly reduced in the IMN group. Moreover, the mean Bishop score and the mean change in Bishop score were significantly increased in the IMN group. Pertaining to drug-related maternal side effect outcomes, the pooled analysis showed that the rates of headache, palpitations, nausea and flushing were significantly lower in the IMN group. Pertaining to neonatal outcomes, the pooled analysis showed no significant difference between the two groups in terms of the rates of neonatal intensive care unit admission, neonatal death, fetal distress, meconium-stained water, Apgar score < 7 at 1 and 5 min, and mean Apgar score at 1 and 5 min. CONCLUSION: IMN correlated with several obstetric-related maternal outcomes. IMN was not associated with adverse neonatal outcomes, but was associated with substantial drug-related maternal side effects.

Characterization of the Interactions between an Unassociated Cationic Pyrene-Labeled Gemini Surfactant and Anionic Sodium Dodecyl Sulfate.

The gemini surfactant PyO-3-12, made of two dimethylammonium bromides joined by a propyl linker and bearing a dodecyl pendant on one side and a 1-pyrenemethoxyhexyl group on the other side, was employed to probe the interactions between positively charged PyO-3-12 and negatively charged sodium dodecyl sulfate (SDS). PyO-3-12 was selected for its ability to respond to the polarity of its local environment through the fluorescence intensity ratio I1/I3 of the first-to-third fluorescence peaks of the pyrene monomer and the local pyrene concentration [Py]loc through the IE/IM ratio of the pyrene excimer-to-pyrene monomer fluorescence intensity. Furthermore, analysis of the fluorescence decays of aqueous solutions of PyO-3-12 and SDS yielded a measure of the internal dynamics, local concentration, and state (associated vs unassociated) of PyO-3-12 in solution. By following these parameters for aqueous solutions prepared with a constant PyO-3-12 concentration of either 1, 4, or 16 µM and SDS concentrations ranging from 0 to 200 mM, six SDS concentration regimes were identified to describe the interactions between PyO-3-12 and SDS in pure water. Sharp transitions of the parameters describing the fluorescence of pyrene marked the boundaries between the different regimes. Perhaps the most important transition was the one defining the formation of the PyO-3-12/SDS aggregates, which was completed at the equicharge point, implying that they were constituted of 1 meq of PyO-3-12 and 2 meq of SDS. The low I1/I3 ratio obtained for the PyO-3-12/SDS aggregates suggested that they were multilamellar aggregates, which would shield the pyrenyl labels from polar water. The formation of these multilamellar aggregates was confirmed by transmission electron microscopy (TEM), which demonstrated the existence of multilamellar vesicles, whose presence increased with decreasing PyO-3-12 concentration. This study suggests that the combination of pyrene excimer formation and TEM provides an interesting experimental means to probe the assemblies generated from oppositely charged surfactants at surfactant concentrations, which are much lower than their critical micelle concentration.

Cyto-Genotoxic and Transcriptomic Alterations in Human Liver Cells by Tris (2-Ethylhexyl) Phosphate (TEHP): A Putative Hepatocarcinogen.

Tris (2-ethylhexyl) phosphate (TEHP) is an organophosphate flame retardant (OPFRs) which is extensively used as a plasticizer and has been detected in human body fluids. Contemporarily, toxicological studies on TEHP in human cells are very limited and there are few studies on its genotoxicity and cell death mechanism in human liver cells (HepG2). Herein, we find that HepG2 cells exposed to TEHP (100, 200, 400 µM) for 72 h reduced cell survival to 19.68%, 49.83%, 58.91% and 29.08%, 47.7% and 57.90%, measured by MTT and NRU assays. TEHP did not induce cytotoxicity at lower concentrations (5, 10, 25, 50 µM) after 24 h and 48 h of exposure. Flow cytometric analysis of TEHP-treated cells elevated intracellular reactive oxygen species (ROS), nitric oxide (NO), Ca++ influx and esterase levels, leading to mitochondrial dysfunction (ΔΨm). DNA damage analysis by comet assay showed 4.67, 9.35, 13.78-fold greater OTM values in TEHP (100, 200, 400 µM)-treated cells. Cell cycle analysis exhibited 23.1%, 29.6%, and 50.8% of cells in SubG1 apoptotic phase after TEHP (100, 200 and 400 µM) treatment. Immunofluorescence data affirmed the activation of P53, caspase 3 and 9 proteins in TEHP-treated cells. In qPCR array of 84 genes, HepG2 cells treated with TEHP (100 µM, 72 h) upregulated 10 genes and downregulated 4 genes belonging to a human cancer pathway. Our novel data categorically indicate that TEHP is an oxidative stressor and carcinogenic entity, which exaggerates mitochondrial functions to induce cyto- and genotoxicity and cell death, implying its hepatotoxic features.

Tris(2-butoxyethyl) phosphate (TBEP): A flame retardant in solid waste display hepatotoxic and carcinogenic risks for humans.

Recent reports have confirmed that tris(2-butoxyethyl) phosphate (TBEP), an organophosphorous flame retardants (OPFRs), profoundly detected in the dust from solid waste (SW), e-waste dumping sites, landfills, and wastewater treatment facilities. Herein, we evaluated the hepatotoxic and carcinogenic potential of TBEP in human liver cells (HepG2). HepG2 cells exhibited cytotoxicity after 3 days of exposure, especially at greater concentrations (100-400 µM). TBEP induced severe DNA damage and cell cycle disturbances that trigger apoptosis in HepG2. TBEP treated cells showed an elevated level of esterase, nitric oxide (NO), reactive oxygen species (ROS), and influx of Ca2+ in exposed cells. Thereby, causing oxidative stress and proliferation inhibition. TBEP exposed HepG2 cells exhibited dysfunction in mitochondrial membrane potential (ΔΨm). Immunofluorescence analysis demonstrated cytoplasmic and nucleolar localization of DNA damage (P53) and apoptotic (caspase 3 and 9) proteins in HepG2 grown in the presence of TBEP for 3 days. Within the cohort of 84 genes of cancer pathway, 10 genes were upregulated and 3 genes were downregulated. The transcriptomic and toxicological data categorically emphasize that TBEP is hepatotoxic, and act as a putative carcinogenic agent. Thereby, direct or indirect ingestion of TBEP containing dusts by workers involved in handling and disposal of SW, as well as residents living nearby the disposal areas are prone to its adverse health risks.

Organophosphorus Flame Retardant TDCPP Displays Genotoxic and Carcinogenic Risks in Human Liver Cells.

Tris(1,3-Dichloro-2-propyl)phosphate (TDCPP) is an organophosphorus flame retardant (OPFR) widely used in a variety of consumer products (plastics, furniture, paints, foams, and electronics). Scientific evidence has affirmed the toxicological effects of TDCPP in in vitro and in vivo test models; however, its genotoxicity and carcinogenic effects in human cells are still obscure. Herein, we present genotoxic and carcinogenic properties of TDCPP in human liver cells (HepG2). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and neutral red uptake (NRU) assays demonstrated survival reduction in HepG2 cells after 3 days of exposure at higher concentrations (100-400 µM) of TDCPP. Comet assay and flow cytometric cell cycle experiments showed DNA damage and apoptosis in HepG2 cells after 3 days of TDCPP exposure. TDCPP treatment incremented the intracellular reactive oxygen species (ROS), nitric oxide (NO), Ca2+ influx, and esterase level in exposed cells. HepG2 mitochondrial membrane potential (ΔΨm) significantly declined and cytoplasmic localization of P53, caspase 3, and caspase 9 increased after TDCPP exposure. qPCR array quantification of the human cancer pathway revealed the upregulation of 11 genes and downregulation of two genes in TDCPP-exposed HepG2 cells. Overall, this is the first study to explicitly validate the fact that TDCPP bears the genotoxic, hepatotoxic, and carcinogenic potential, which may jeopardize human health.

Synthesis and Characterization of a Pyrene-Labeled Gemini Surfactant Sensitive to the Polarity of Its Environment.

The cationic gemini surfactant PyO-3-12 was designed to include two dimethyl ammonium groups, one dodecyl tail, and 1-pyrenemethyl hexyl ether tail into the structure of the surfactant. The pyrenyl label ensured that the fluorescence of pyrene could be employed to probe the behavior of PyO-3-12 at the molecular level. The introduction of the oxygen atom in the ß-position to pyrene was found to be critical for restoring the sensitivity of the pyrenyl label to the polarity of its environment. The properties of PyO-3-12 were characterized in water by surface tension and a fluorescence methodology that involved the global model-free analysis (MFA) of the pyrene monomer and excimer fluorescence decays to provide quantitative information about the state (unassociated-vs-aggregated) of PyO-3-12. The MFA was combined with a fluorescence quenching study with 2,6-dinitrotoluene to determine the size of the PyO-3-12 micelles. PyO-3-12 was found to behave like a typical gemini surfactant, exhibiting a critical micelle concentration (CMC) of 0.38 (±0.05) mM and an aggregation number (Nagg) equal to 23 (±2). Besides allowing PyO-3-12 to probe the polarity of its environment, the oxygen atom in the ß-position next to pyrene brought some pyrenyl labels closer to the interface between the micellar interior and the aqueous phase, in a process that increased the effective volume of the hydrophobic part of PyO-3-12. This led to an increase in the packing parameter of PyO-3-12 and, consequently, an increase in Nagg compared to the Nagg value of 14 (±0.2) obtained for Py-3-12, a gemini surfactant, whose chemical structure was similar to that of PyO-3-12 but without the oxygen in the ß-position to pyrene. The methodology described in this study to prepare and characterize pyrene-labeled surfactants is general and can be applied to study any pyrene-labeled surfactant and its interactions with oppositely charged macromolecules.

Determination of the Aggregation Number of Pyrene-Labeled Gemini Surfactant Micelles by Pyrene Fluorescence Quenching Measurements.

A cationic gemini surfactant referred to as Py-3-12 and composed of two alkylated diammonium bromide head groups, a propyl spacer, and dodecyl and 1-pyrenehexyl hydrophobic tails was synthesized. Its critical micellar concentration (CMC) was determined to equal 0.15 (±0.02) mM by surface tension and time-resolved fluorescence measurements. The state of the pyrene molecules, whether they were incorporated inside the Py-3-12 micelles or unassociated in the aqueous solution, was determined by applying the global model-free analysis (MFA) to the fluorescence decays acquired with Py-3-12 aqueous solutions. The unassociated Py-3-12 surfactants emitted as pyrene monomers and showed a long fluorescence lifetime. The excited pyrenyl groups located inside Py-3-12 micelles formed an excimer by a rapid encounter with a ground-state pyrene with an average rate constant equal to 0.69 (±0.06) ns-1. After having the photophysical properties of Py-3-12 in aqueous solution characterized, the number (Nagg) of surfactants per micelle was determined by conducting quenching experiments with dinitrotoluene (DNT). Although DNT is fairly hydrophobic, it was found to partition itself between the Py-3-12 micelles and the aqueous phase. Fluorescence quenching experiments performed on the pyrene monomer and excimer generated by the Py-3-12 aqueous solutions yielded the concentration ([Q]b) of DNT bound to the Py-3-12 micelles and the average number ⟨n⟩d of DNT quenching an excimer by diffusive encounters. A combination of steady-state and time-resolved fluorescence quenching experiments on the excimer yielded the number (⟨n⟩s) of DNT molecules that were bound to the micelles and quenched the excimer in a static manner. A plot of the sum ⟨n⟩d + ⟨n⟩s as a function of [Q]b yielded an Nagg value of 14.0 (±0.2) Py-3-12 units per micelle. This study represents the first example in the literature where Nagg is determined for a micelle, where each surfactant molecule is labeled with pyrene.

Tris(2-chloroethyl) Phosphate (TCEP) Elicits Hepatotoxicity by Activating Human Cancer Pathway Genes in HepG2 Cells.

Tris(2-chloroethyl) phosphate (TCEP) is one of the organophosphorus flame retardants (OPFRs) used in consumer commodities and have been detected in human body fluids. Research on TCEP-induced transcriptomic alterations and toxicological consequences in liver cells is still lacking. Herein, human hepatocellular (HepG2) cells were treated with 100, 200, and 400 µM TCEP for 3 days to quantify hepatotoxicity by MTT, NRU, and comet assays. Apoptosis, mitochondrial membrane potential (ΔΨm), oxidative stress, and Ca2+ influx were measured by flow cytometry. A qPCR array was employed for transcriptomic analysis. MTT and NRU data showed 70.92% and 75.57% reduction in cell survival at 400 µM. In addition, 20-fold greater DNA damage was recorded at 400 µM. Cell cycle data showed 65.96% subG1 apoptotic peak in 400 µM treated cells. An elevated level of oxidative stress, esterase, Ca2+ influx, and ΔΨm dysfunction were recorded in TCEP-treated cells. Out of 84 genes, the qPCR array showed upregulation of 17 genes and downregulation of 10 key genes belonging to human cancer pathways. Our study endorses the fact that TCEP possesses hepatotoxic potential at higher concentrations and prolonged exposure. Hence, TCEP may act as a cancer-inducing entity by provoking the gene network of human cancer pathways.

Preoperative Leukocytosis as a Prognostic Marker in Endometrioid-Type Endometrial Cancer: A Single-Center Experience from Saudi Arabia.

INTRODUCTION: Only a few studies (n=5) have focused on the importance of preoperative high white blood cell (WBC) count (leukocytosis) as a prognostic marker in patients with endometrial cancer (EC). Nevertheless, more related studies are needed to solidly corroborate these findings. To the best of our knowledge, no such study has been conducted in the Gulf region and Saudi Arabia in particular. METHODS: A retrospective cross-sectional study was conducted at King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia. The medical records of 130 patients with endometrioid-type EC were reviewed for clinico-pathological factors (that is, age, tumor stage, endometrioid grade, myometrial invasion depth, lymphovascular space involvement and recurrence) and survival outcomes. Survival outcomes included disease-free survival (DFS) and overall survival (OS). Leukocytosis was defined as a WBC count level >10 x 103 cells/uL. Chisquare test was used for univariate analysis of categorical data. Survival analyses of DFS and OS were calculated according to the Kaplan-Meier estimates method and compared by using two-tailed log-rank test. Univariate and multivariate analyses of survival were performed using Cox proportional hazards model. Statistical significance was regarded as a p value <0.05. RESULTS: The mean age was 59 ± 10.5 years (range: 36-99). The overall mean preoperative WBC count was 7.7 ± 2.4 x 103 cells/uL (range: 2.7-17 x 103). The frequency of preoperative leukocytosis was 18.5% (n=24). Patients with preoperative leukocytosis have statistically significant higher rates of advanced FIGO stage III-IV disease (p=0.007) and positive tumor recurrence (p=0.009) than patients with normal preoperative WBC count (chisquare test). Patients with preoperative leukocytosis have a higher statistically significant probability of developing recurrence than patients with preoperative normal WBC count (29.4 vs. 11.8%, p=0.008, log-rank test). Patients with preoperative leukocytosis have statistically significant lower mean DFS (58.3 ± 6.9 vs. 67.9 ± 2.3 months, p=0.015) and 5-year DFS rate (66.7 vs. 86.8%, p=0.015) than patients with normal preoperative WBC counts (log-rank test). However, there were no statistically significant differences between patients with preoperative leukocytosis and normal WBC counts in terms of mean OS (73.8 ± 4.5 vs. 79.3 ± 2.1, p=0.581) and 5-year OS rate (87.5 vs. 91.5%, p=0.581), respectively (log-rank test). Multivariate analyses using Cox proportional hazards model failed to significantly demonstrate preoperative WBC count as an independent prognostic factor of DFS and OS (log-rank test, p>0.05). CONCLUSION: Preoperative leukocytosis is not rare in patients with endometrioid-type EC. Besides, preoperative leukocytosis is correlated with poor tumor FIGO stage, higher cumulative incidence of relapse and poor DFS in the univariate analysis. Our study suggests that preoperative leukocytosis may identify high-risk patients who may require more intensified therapy in terms of aggressive debulking and/or perioperative chemotherapy.

Organophosphorus flame retardant (tricresyl phosphate) trigger apoptosis in HepG2 cells: Transcriptomic evidence on activation of human cancer pathways.

Tricresyl phosphate (TCP) is one of the organophosphorus flame retardants (OPFRs) used as plasticizer in consumer products and mixed as a lubricant in commercial jet engine oil, reportedly induce neurotoxicity and aerodynamic syndrome. No studies have been attempted so far on TCP to induce hepatotoxicity in human cells. This study for the first time confirms the hepatotoxic potential and activation of cancer pathways in TCP treated human hepatocellular cells (HepG2). MTT and NRU data showed 39.3% and 49.85% decline in HepG2 survival when exposed to the highest concentration of TCP (400 µM) for 3 days. Comet assay showed 27.1-fold greater DNA damage in cells treated with TCP (400 µM). Flow cytometric analysis revealed an upsurge in the intracellular reactive oxygen species (ROS) and nitric oxide (NO) production in cells, affirming oxidative stress. TCP (400 µM) exposure resulted in 27% reduction in Rh123 fluorescence, indicating dysfunction of mitochondrial membrane potential (ΔΨm). Cell cycle analysis exhibited 62.53% cells in the subG1 apoptotic phase after TCP (400 µM) treatment, also a massive increase in Ca2+ influx validate the on-set of apoptosis in cells. Immunofluorescence of TCP exposed cells showed activation of p53, caspase3, caspase9 reaffirming the involvement of mitochondrial-dependent intrinsic apoptotic signaling. qPCR array of 84 genes unravel the transcriptomic alterations in HepG2 cells after TCP treatment. mRNA transcripts of ATP5A1, GADD45A, IGFBP5, SOD1, STMN1 genes were prominently upregulated providing candid evidence on TCP mediated activation of human cancer pathways to orchestrate the apoptotic death of HepG2 cells, specifying hepatotoxic potential of TCP.

Quantum dot-based molecular beacon to monitor intracellular microRNAs.

Fluorescence monitoring of endogenous microRNA (miRNA or miR) activity related to neuronal development using nano-sized materials provides crucial information on miRNA expression patterns in a noninvasive manner. In this study, we report a new method to monitor intracellular miRNA124a using quantum dot-based molecular beacon (R9-QD-miR124a beacon). The R9-QD-miR124a beacon was constructed using QDs and two probes, miR124a-targeting oligomer and arginine rich cell-penetrating peptide (R9 peptide). The miR124a-targeting oligomer contains a miR124a binging sequence and a black hole quencher 1 (BHQ1). In the absence of target miR124a, the R9-QD-miR124a beacon forms a partial duplex beacon and remained in quenched state because the BHQ1 quenches the fluorescence signal of the R9-QD-miR124a beacon. The binding of miR124a to the miR124a binding sequence of the miR124a-targeting oligomer triggered the separation of the BHQ1 quencher and subsequent signal-on of a red fluorescence signal. Moreover, enhanced cellular uptake was achieved by conjugation with the R9 peptide, which resulted in increased fluorescent signal of the R9-QD-miR124a beacons in P19 cells during neurogenesis due to the endogenous expression of miR124a.

Gd-Complexes of New Arylpiperazinyl Conjugates of DTPA-Bis(amides): Synthesis, Characterization and Magnetic Relaxation Properties.

Two new DTPA-bis(amide) based ligands conjugated with the arylpiperazinyl moiety were synthesized and subsequently transformed into their corresponding Gd(III) complexes 1 and 2 of the type [Gd(L)H2O]·nH2O. The relaxivity (R1) of these complexes was measured, which turned out to be comparable with that of Omniscan®, a commercially available MRI contrast agent. The cytotoxicity studies of these complexes indicated that they are non-toxic, which reveals their potential and physiological suitability as MRI contrast agents. All the synthesized ligands and complexes were characterized with the aid of analytical and spectroscopic methods, including elemental analysis, 1H-NMR, FT-IR, XPS and fast atom bombardment (FAB) mass spectrometry.