Integration of LC/MS, NMR and Molecular Docking for Profiling of Bioactive Diterpenes from Euphorbia mauritanica L. with in Vitro Anti-SARS-CoV-2 Activity.

In spite of tremendous efforts exerted in the management of COVID-19, the absence of specific treatments and the prevalence of delayed and long-term complications termed post-COVID syndrome still urged all concerned researchers to develop a potent inhibitor of SARS-Cov-2. The hydromethanolic extracts of different parts of E. mauritanica were in vitro screened for anti-SARS-Cov-2 activity. Then, using an integrated strategy of LC/MS/MS, molecular networking and NMR, the chemical profile of the active extract was determined. To determine the optimum target for these compounds, docking experiments of the active extract's identified compounds were conducted at several viral targets. The leaves extract showed the best inhibitory effect with IC50 8.231±0.04 µg/ml. The jatrophane diterpenes were provisionally annotated as the primary metabolites of the bioactive leaves extract based on multiplex of LC/MS/MS, molecular network, and NMR. In silico studies revealed the potentiality of the compounds in the most active extract to 3CLpro, where compound 20 showed the best binding affinity. Further attention should be paid to the isolation of various jatrophane diterpenes from Euphorbia and evaluating their effects on SARS-Cov-2 and its molecular targets.

Characterization of undescribed melanoma inhibitors from Euphorbia mauritanica L. cultivated in Egypt targeting BRAFV600E and MEK 1 kinases via in-silico study and ADME prediction.

Three undescribed diterpenes including two ent-abietanes, euphomauritanol A, and euphomauritanol B, and one jatrophane, euphomauritanophane A, in addition to eight previously described metabolites were isolated from the MeOH-CH2Cl2 (1:1) extract of the Euphorbia mauritanica. The chemical structures of isolates were established based on the spectroscopic means including FT-IR, HRMS, 1D and 2D NMR. The absolute stereochemistry of the undescribed diterpenes was deduced by experimental and calculated TDDFT-electronic circular dichroism (ECD). The anti-proliferative effects of the isolated diterpenes were evaluated against B16-BL6, Hep G2, and Caco-2. The euphomauritanol A, euphomauritanol B, and euphomauritanophane A significantly inhibited the growth of murine melanoma B16-BL6 cell lines with IC50 10.28, 20.22, and 38.81 µM, respectively with no responses against the other cells. These activities were rationalized by molecular docking of the active compounds in BRAFV600E and MEK1 active sites. Moreover, the in-silico pharmacokinetics predictions by Swiss ADME revealed that the active compounds possessed favorable oral bioavailability and drug-likeness properties.

A laboratory study to test the responses of human dental pulp stem cells to extracts from three dental pulp capping biomaterials.

AIM: This laboratory study aimed to investigate the effects of three endodontic biomaterials; MTA-HP, iRoot-BP-Plus and ACTIVA on the proliferation, adhesion and osteogenic differentiation of human Dental Pulp Stem Cells (hDPSCs). METHODOLOGY: The hDPSCs were isolated from the dental pulps of 21 patients scheduled for surgical extraction of their impacted third molars. The MTT assay was used for assessing cellular proliferation. Ninety-six-well plates were used and the experiment was repeated four times under the same condition and the assay was done in triplicate. Four groups were assigned in which the hDPSCs were cultured in complete media only and considered as negative control. Whilst in the 2nd , 3rd and 4th groups, the cells were treated with CM supplemented with 1.5 µl MTA-HP (CM-MTA, iRoot-BP-Plus (CM-BP), and ACTIVA(CM-AC) extracts, respectively. Attachment adhesion and growth morphology of hDPSCs were observed using SEM and the osteogenic differentiation assay was evaluated by Alizarin red stain test (ARS). The data of proliferation and osteogenic differentiation were analysed using two-way ANOVA followed by Tukey's post hoc multiple comparison test. A p-value < 0.05 was considered significant to analyse the differences amongst the means of groups. RESULTS: Both CM-MTA and CM-BP groups were associated with a significant increase in hDPSC proliferation in comparison with CM-AC and CM groups (p = 0.001). hDPSCs exhibited a greater cellular attachment to iRoot-BP-Plus surfaces followed by MTA-HP, whilst less attachment was observed in the ACTIVA group. Moreover, at day 7 there was a significant difference in formation of mineralizing nodules; CM-BP, CM-MTA and CM-AC groups respectively (p = 0.001). Whilst there was no significance of difference between CM-AC and CM groups (p > 0.05). CONCLUSIONS: In a laboratory setting, ACTIVA, MTA-HP and iRoot-BP-Plus promoted hDPSCs proliferation, mineralization and attachment, which may explain their in-situ success as endodontic biomaterials.

Novel coumarin-6-sulfonamides as apoptotic anti-proliferative agents: synthesis, in vitro biological evaluation, and QSAR studies.

Herein, we report the synthesis of different novel sets of coumarin-6-sulfonamide derivatives bearing different functionalities (4a, b, 8a-d, 11a-d, 13a, b, and 15a-c), and in vitro evaluation of their growth inhibitory activity towards the proliferation of three cancer cell lines; HepG2 (hepatocellular carcinoma), MCF-7 (breast cancer), and Caco-2 (colon cancer). HepG2 cells were the most sensitive cells to the influence of the target coumarins. Compounds 13a and 15a emerged as the most active members against HepG2 cells (IC50 = 3.48 ± 0.28 and 5.03 ± 0.39 µM, respectively). Compounds 13a and 15a were able to induce apoptosis in HepG2 cells, as assured by the upregulation of the Bax and downregulation of the Bcl-2, besides boosting caspase-3 levels. Besides, compound 13a induced a significant increase in the percentage of cells at Pre-G1 by 6.4-folds, with concurrent significant arrest in the G2-M phase by 5.4-folds compared to control. Also, 13a displayed significant increase in the percentage of annexin V-FITC positive apoptotic cells from 1.75-13.76%. Moreover, QSAR models were established to explore the structural requirements controlling the anti-proliferative activities.

Baseline evaluation for natural radioactivity level and radiological hazardous parameters associated with processing of high grade monazite.

Egyptian monazite is a promising resource and investment attractive for production of valuable metals of industrial or nuclear interest such as rare earth elements (REEs), thorium (Th) and uranium (U). The study was focused to establish a baseline framework in viewpoint of radiation protection for the workers in production of REEs from high-grade monazite treated by sodium hydroxide (NaOH) solutions. Radiological hazard indices (cancer, gonadal and other risks) were evaluated, due to emissions (α-, ß- and γ-radiations) of radium-isotopes (228Ra, 226Ra, 223Ra) and lead (210Pb). The values of the estimated radiological hazard indices were higher than the permissible safe limits, worldwide average and varied with those reported in other countries. It was found that more than 70% of radioactivity and radiological hazardous indices resulted from emissions of 228Ra, while the rest was attributed to 226Ra, 223Ra and 210Pb. Therefore, processing of the Egyptian monazite can cause a significant radiological impact on workers through external exposure from γ-radiations and/or internal exposure through inhalation or ingestion airborne contaminated by the radionuclides. Thus, the results recommended that protection rules could be considered to prevent the radiation hazards associated with the production of the REEs from the high grade monazite attacked by caustic method.

Catalase improves motility, vitality and DNA integrity of cryopreserved human spermatozoa.

Cryopreservation of human spermatozoa offers a pre-therapeutic possibility of preserving progenity in patients with testicular tumours. We aimed to investigate effects of cryopreservation and addition of catalase on sperm motility, vitality and DNA integrity in fresh and swim-up spermatozoa. Semen samples were collected from 50 fertile men. Each sample was divided into two parts. First part was subdivided into two equal aliquots: both cryopreserved with and without catalase. The second part was subdivided into two equal aliquots: both processed by swim up and then cryopreserved with or without catalase. Semen analyses, sperm vitality and sperm DNA integrity were performed. Sperm concentration showed significant decrease while percentage of progressive motility, sperm vitality and % of DNA damage showed significant increase in processed and cryopreserved processed samples when compared with fresh and cryopreserved fresh samples. There was no significant difference in sperm concentration while there was significant increase in % of progressive motility and sperm vitality and % of DNA damage showed significant decrease in samples with catalase when compared with samples without catalase (either fresh or processed). Catalase supplementation (fresh and processed) during cryopreservation results in better post-thawing percentage of progressive motility and percentage of sperm vitality and improved DNA integrity.

High performance liquid chromatography, thin layer chromatography and spectrophotometric studies on the removal of biogenic amines from some Egyptian foods using organic, inorganic and natural compounds.

This work has been carried out to investigate the conditions which lead to removal of the biogenic amines through the model system. Also, the main goal of this research work is trying to remove biogenic amines; histamine and tyramine, from some Egyptian foods such as tomato, strawberry, banana and mango to prevent their allergy effect. Histamine and tyramine have been affected by pyrogallol, catechol, starch, ascorbic and chlorogenic acids at different levels with different conditions. Some natural additives like glucose, spices, milk, vanillin, starch, orange juice, ascorbic and citric acids, showed an effective effect on disappearance of histamine and tyramine. By studying the effect of some additives on biogenic amines, it was found that tomato showed a decrease in histamine and tyramine concentrations by adding spices. Strawberry and banana showed a clear decrease in histamine and tyramine concentrations by treating them with ascorbic acid. Treating mango by milk led to increase of histamine level while milk with chocolate increases both histamine and tyramine concentrations.

Alterations of mononuclear inflammatory cells, CD4/CD8+ T cells, interleukin 1beta, and tumour necrosis factor alpha in the bronchoalveolar lavage fluid, peripheral blood, and skin of patients with systemic sclerosis.

BACKGROUND: Systemic sclerosis (SSc) is a multisystem disease with underlying immune mechanisms. AIMS: To investigate the clinicopathological characteristics of the lesions; immunological alterations in the bronchoalveolar lavage fluid (BALF), peripheral blood, and skin; and correlations between the clinicopathological characteristics and immunological alterations in SSc. MATERIALS/METHODS: Skin biopsies, BALF, and peripheral blood samples were obtained from 19 patients (18 women, one man) with SSc and six age and sex matched healthy controls (HCs). Mononuclear inflammatory cells (MICs), CD4/CD8 cells, tumour necrosis factor alpha (TNFalpha), and interleukin 1beta (IL1-1beta) concentrations were examined in all samples using histological methods, enzyme linked immunosorbent assay, and immunoperoxidase staining. RESULTS: The mean (SD) age of the patients with SSc was 34.8 (2.6) years. Proteinuria, positive rheumatoid factor, and C reactive protein were seen in 15.8%, 26.3%, and 26.3% of patients, respectively. Compared with HCs, there were significantly higher: total MICs (macrophages, lymphocytes), neutrophils, and eosinophils in BALF, blood, and skin (all p<0.05); cytokine concentrations in BALF (TNFalpha, p<0.001; IL-1, p<0.01) and peripheral blood (p<0.01 and p<0.05); and CD8/CD4+ T cells in peripheral blood (p<0.05). Compared with HCs, lesional skin had significantly higher histiocyte cell counts (p<0.05), lower lymphocyte counts (p<0.05), and higher CD4/CD8 ratios (p<0.001). There were significant correlations between cytokine concentrations and CD8+ T cells and forced vital capacity (p<0.001 and p<0.01, respectively). CONCLUSIONS: MICs, CD4/CD8+ cells, and cytokines are altered in SSc. These alterations correlated with the underlying disease process and therefore may have pathogenic, modulatory, and potential prognostic roles in SSc.

A benzoquinone and flavonoids from Cyperus alopecuroides.

A benzoquinone, named alopecuquinone, was isolated from the ethanol extract of the inflorescences of Cyperus alopecuroides. Its structure was primarily elucidated by spectroscopic analysis including 1H, 13C NMR, APT, HMQC, 1H-1H COSY and CIMS. The known flavonoids, vicenin 2, orientin, diosmetin, quercetin 3,3'-dimethyl ether and its 3,4'-dimethyl ether, were also isolated and characterized. The ethanol extract of the plant material showed moderate estrogenic activity using a strain of Saccharomyces cerevisiae.

Inhibitory effects of thymoquinone against 20-methylcholanthrene-induced fibrosarcoma tumorigenesis.

The potential antitumor effect of thymoquinone (TQ), the main constituent of the volatile oil of Nigella sativa seed, on fibrosarcoma induced by 20-methylcholanthrene (MC) in male Swiss albino mice was investigated in vivo and in vitro. Administration of TQ (0.01% in drinking water) I week before and after MC treatment significantly inhibited the tumor incidence and tumor burden by 43% and 34%, respectively, compared with the results in the group receiving MC alone. Moreover, TQ delayed the onset of MC-induced fibrosarcoma tumors that appeared at 12 weeks and produced less MC-induced mortality. Lipid peroxide accumulation, decreased glutathione (GSH) content, and decreased activities of glutathione S-transferase (GST) and quinone reductase (QR) were observed in the liver of MC-induced tumor-bearing mice. TQ alone showed a significant induction in the enzyme activities of hepatic GST and QR. Mice treated with TQ along with MC showed reduction in hepatic lipid peroxides and increased GSH content and increased enzyme activities of GST and QR as compared to results of the control group. The in vitro studies showed that TQ inhibited the survival of fibrosarcoma cells with IC50 of 15 microM. Conversely, TQ inhibited the incorporation of [3H] thymidine in fibrosarcoma cells with IC50 of microM. Our data indicate the potential of TQ as a powerful chemopreventive agent against MC-induced fibrosarcoma tumors. The possible modes of action of TQ may be through its antioxidant activity and interference with the DNA synthesis coupled with enhancement of detoxification processes.

Cartilage proteoglycan aggrecan as a predictor of joint damage in juvenile rheumatoid arthritis.

Aggrecan was measured in the sera of 31 children with juvenile rheumatoid arthritis and in the synovial fluid of 10 of them. Patients were evaluated at baseline and 3 months later. Radiographs were repeated also after 1 year. As comparison, 15 apparently healthy children with no disease and 10 children with arthritis due to other collagen vascular diseases were studied. Baseline serum aggrecan was significantly higher in juvenile rheumatoid arthritis patients compared to controls and other patients. On re-evaluation, a significant drop in serum aggrecan from baseline values coincided with a significant drop in clinical and laboratory indices of active inflammation. Serum aggrecan can help to assess the extent of cartilage destruction and is useful as a prognostic tool to predict joint damage in patients with juvenile rheumatoid arthritis.

Cartilage proteoglycan aggrecan as a predictor of joint damage in juvenile rheumatoid arthritis

Aggrecan was measured in the sera of 31 children with juvenile rheumatoid arthritis and in the synovial fluid of 10 of them. Patients were evaluated at baseline and 3 months later. Radiographs were repeated also after 1 year. As comparison, 15 apparently healthy children with no disease and 10 children with arthritis due to other collagen vascular diseases were studied. Baseline serum aggrecan was significantly higher in juvenile rheumatoid arthritis patients compared to controls and other patients. On re-evaluation, a significant drop in serum aggrecan from baseline values coincided with a significant drop in clinical and laboratory indices of active inflammation. Serum aggrecan can help to assess the extent of cartilage destruction and is useful as a prognostic tool to predict joint damage in patients with juvenile rheumatoid arthritis

Trauma patients 75 years and older: long-term follow-up results justify aggressive management.

BACKGROUND: Long-term survival rate and functional status after trauma for one of the fastest growing segments of the population, patients 75 years and older, is poorly documented. METHODS: Trauma patients 75 years and older who were discharged from our Level I trauma center between June 1988 and July 1992 (n = 279) were contacted by mail or phone. Public death records were used to identify patients who had died. A stepwise logistic regression analysis was performed to determine predictors of poor outcome (death within 6 months). Main outcome measures included mortality and self-assessed functional status. RESULTS: A minimum 4-year follow-up was obtained for 81% of the 279 study patients. The mean follow-up period was 5.4 +/- 1.1 years. Mean age at time of injury was 81 +/- 5 years (range, 75-101 years); mean Injury Severity Score was 9.4 +/- 7.7. At follow-up, 132 patients (47%) had died, 93 patients (33%) were contacted, and 54 patients (19%) could not be located. Twelve percent of patients survived less than 6 months after discharge. Poor survival was predicted by preexisting diseases (dementia, p = 0.001; hypertension, p = 0.02; and chronic obstructive pulmonary disease, p = 0.05) and not by age or severity of injury. The mean age of patients still living was 85 +/- 3.9 years (range, 79-99 years), and 77 of 93 patients were living in an independent setting (33 alone, 44 with spouse or family); of these, 57% reported no difficulties in performing 12 of 14 activities of daily living. CONCLUSION: Despite higher than expected mortality after discharge, aggressive management of trauma patients 75 years and older is justified by the favorable long-term outcome.

Induction of tumor necrosis factor in severe acute pancreatitis and its subsequent reduction after hepatic passage.

BACKGROUND: Tumor necrosis factor (TNF) is rapidly gaining recognition as one of the early, critical mediators of several inflammatory conditions, most notably endotoxic shock. The purposes of this study were to determine whether TNF levels are raised in severe acute pancreatitis, thus pointing to its role as a potential mediator of the inflammatory process, and to determine the possible sites of production and uptake. METHODS: TNF levels were measured during a 2-hour period in a rat model of acute pancreatitis by using an antegrade infusion of artificial bile. TNF levels were measured with a bioassay. RESULTS: TNF levels increased proportionately with time and serum amylase level, reaching a mean value of 2700 pg/ml at 2 hours compared with sham operated rats (mean, 125 pg/ml) (p < 0.001). TNF levels in nonoperated controls were undetectable. These measurements were found to be independent of endotoxin production. In addition, selective sampling from the portal vein, hepatic vein, and femoral artery showed hepatic degradation of TNF (p < 0.005), indicating that the liver may play an important role in protecting the host from multiple organ failure. CONCLUSIONS: Our results showed that TNF levels are elevated in acute pancreatitis and may suggest a role for this cytokine in the pathogenesis of the disease.