Increased therapeutic potential of an experimental anti-mitotic inhibitor SB715992 by genistein in PC-3 human prostate cancer cell line

BACKGROUND: Kinesin spindle proteins (KSP) are motor proteins that play an essential role in mitotic spindle formation. HsEg5, a KSP, is responsible for the formation of the bipolar spindle, which is critical for proper cell division during mitosis. The function of HsEg5 provides a novel target for the manipulation of the cell cycle and the induction of apoptosis. SB715992, an experimental KSP inhibitor, has been shown to perturb bipolar spindle formation, thus making it an excellent candidate for anti-cancer agent. Our major objective was a) to investigate the cell growth inhibitory effects of SB715992 on PC-3 human prostate cancer cell line, b) to investigate whether the growth inhibitory effects of SB715992 could be enhanced when combined with genistein, a naturally occurring isoflavone and, c) to determine gene expression profile to establish molecular mechanism of action of SB715992. METHODS: PC-3 cells were treated with varying concentration of SB715992, 30 μM of genistein, and SB715992 plus 30 μM of genistein. After treatments, PC-3 cells were assayed for cell proliferation, induction of apoptosis, and alteration in gene and protein expression using cell inhibition assay, apoptosis assay, microarray analysis, real-time RT-PCR, and Western Blot analysis. RESULTS: SB715992 inhibited cell proliferation and induced apoptosis in PC-3 cells. SB715992 was found to regulate the expression of genes related to the control of cell proliferation, cell cycle, cell signaling pathways, and apoptosis. In addition, our results showed that combination treatment with SB715992 and genistein caused significantly greater cell growth inhibition and induction of apoptosis compared to the effects of either agent alone. CONCLUSION: Our results clearly show that SB715992 is a potent anti-tumor agent whose therapeutic effects could be enhanced by genistein. Hence, we believe that SB715992 could be a novel agent for the treatment of prostate cancer with greater success when combined with a non-toxic natural agent like genistein

Study of the wetting of coke by different pitches

Prebaked anodes are made of dry aggregates (coke, recycled butts and rejected green/baked anodes) and the binder pitch. During the mixing process, the wettability of coke by pitch influences the anode paste behavior. Coke particles (-125 +100 m) were prepared from the same source using two different procedures, and the wettability of cokes by five pitches with different properties was studied using a sessile-drop system at 170ºC. The contact angle results show that the wettability of coke by all pitches follows a similar trend for both cases, but the actual contact angles are different. The cokes and the pitch-coke interfaces were also investigated by the optical and scanning-electron microscopy (SEM) techniques. The image analysis results indicated that the coke particle size distribution depends on the coke preparation, which seems to influence its wettability

Switching to Dolutegravir/Lamivudine Two-Drug Regimen: Durability and Virologic Outcomes by Age, Sex, and Race in Routine US Clinical Care

Gerald Pierone Jr,1 Laurence Brunet,2 Jennifer S Fusco,2 Cassidy E Henegar,3 Supriya Sarkar,3 Jean Van Wyk,4 Vani Vannappagari,3 Michael B Wohlfeiler,5 Gregory P Fusco2 1Department of Adult Primary Care, Whole Family Health Center, Vero Beach, FL, USA; 2Department of Epidemiology, Epividian, Raleigh, NC, USA; 3Epidemiology and Real World Evidence, ViiV Healthcare, Research Triangle Park, NC, USA; 4Global Medical, ViiV Healthcare, Brentford, UK; 5Department of Medicine, AIDS Healthcare Foundation, Miami, FL, USACorrespondence: Laurence Brunet, Epividian, 150 Fayetteville Street, Suite 2300, Raleigh, NC, 27601, USA, Tel +1-919-827-0010, Email [email protected]: Two-drug regimens (2DR) may address drug–drug interactions and toxicity concerns. Dolutegravir/lamivudine (DTG/3TC) 2DR was approved in the US for both treatment-naïve and treatment-experienced individuals with a viral load < 50 copies/mL. This study describes real-world DTG/3TC 2DR treatment outcomes among treatment-experienced individuals, stratified by age, sex, and race.Methods: From the OPERA® cohort, people with HIV with a viral load < 50 copies/mL who switched from a commonly used three-drug regimen to DTG/3TC 2DR as per the label between April 8, 2019 and April 30, 2021 were included. Incidence rates (Poisson regression) for loss of virologic control (first viral load ≥ 50 copies/mL), confirmed virologic failure (2 viral loads ≥ 200 copies/mL or discontinuation after 1 viral load ≥ 200 copies/mL), and DTG/3TC 2DR discontinuation were estimated overall and stratified by age, sex, and race.Results: The 787 individuals included were followed for a median of 13.6 months (IQR: 8.2, 22.3). Confirmed virologic failure occurred in ≤ 5 individuals. Loss of virologic control occurred at a rate of 14.0 per 100 person-years (95% CI: 11.7, 16.8). DTG/3TC 2DR discontinuation occurred at a rate of 17.5 per 100 person-years (95% CI: 15.0, 20.3); 4% discontinued for treatment-related reasons (viremia, adverse diagnosis, side effect, lab abnormality). For all outcomes, incidence rates were comparable across strata of age, sex, and race.Conclusion: This descriptive study demonstrates that DTG/3TC 2DR is an effective and well-tolerated treatment option for people with HIV with a viral load < 50 copies/mL at switch, regardless of their age, sex, or race.Keywords: antiretroviral therapy, cohort, electronic health records, suppressed, viral loa

Assessment of the embryotoxic potential of contaminated sediments using fish embryotoxicity tests for the river Buriganga, Dhaka, Bangladesh

Sediment samples from six different locations of Buriganga River following exposure to Zebrafish (Danio rerio) eggs and larvae displayed prominent effects on both whole sediments and sediment organic extracts. The acute and sublethal effects during 96 h exposure period included (i) a significant (P<0.05) increase in morality and abnormalities in zebrafish eggs and embryos; (ii) a significant (P<0.05) reduction in hatching success and heart rate; (iii) increased frequency of helical tail and lordosis after 96 h exposure to sediment extracts; (iv) developmental delay and yolk sac edema after exposed to whole sediments at 96h exposure period. Chemical analysis showed the increaseds concentrations of heavy metals (Zn, Cr, Cu, Pb, and Cd) in downstream (S1, S2, and S3) compared to upstream (S4, S5, and S6), where some ions such as Cd and Cr exceeded the Environmental Protection Agency’s Threshold Effect Level (EPA TEL). The current study delineates the contamination of extremely toxic compounds in the sediment of Buriganga River, which may initiate toxic effects on the early life stages of fish. Therefore, integrating zebrafish embryo toxicity tests may be crucial for evaluating the sediment quality of polluted rivers

Sphingosine 1-phosphate modulates antigen capture by murine langerhans cells via the S1P2 receptor subtype

Dendritic cells (DCs) play a pivotal role in the development of cutaneous contact hypersensitivity (CHS) and atopic dermatitis as they capture and process antigen and present it to T lymphocytes in the lymphoid organs. Recently, it has been indicated that a topical application of the sphingolipid sphingosine 1-phosphate (S1P) prevents the inflammatory response in CHS, but the molecular mechanism is not fully elucidated. Here we indicate that treatment of mice with S1P is connected with an impaired antigen uptake by Langerhans cells (LCs), the initial step of CHS. Most of the known actions of S1P are mediated by a family of five specific G protein-coupled receptors. Our results indicate that S1P inhibits macropinocytosis of the murine LC line XS52 via S1P2 receptor stimulation followed by a reduced phosphatidylinositol 3-kinase (PI3K) activity. As down-regulation of S1P2 not only diminished S1P-mediated action but also enhanced the basal activity of LCs on antigen capture, an autocrine action of S1P has been assumed. Actually, S1P is continuously produced by LCs and secreted via the ATP binding cassette transporter ABCC1 to the extracellular environment. Consequently, inhibition of ABCC1, which decreased extracellular S1P levels, markedly increased the antigen uptake by LCs. Moreover, stimulation of sphingosine kinase activity, the crucial enzyme for S1P formation, is connected not only with enhanced S1P levels but also with diminished antigen capture. These results indicate that S1P is essential in LC homeostasis and influences skin immunity. This is of importance as previous reports suggested an alteration of S1P levels in atopic skin lesions

Quantum corrections and black hole spectroscopy

In the work \cite{BRM,RBE}, black hole spectroscopy has been successfully reproduced in the tunneling picture. As a result, the derived entropy spectrum of black hole in different gravity (including Einstein's gravity, Einstein-Gauss-Bonnet gravity and Ho\v{r}ava-Lifshitz gravity) are all evenly spaced, sharing the same forms as $S_n=n$, where physical process is only confined in the semiclassical framework. However, the real physical picture should go beyond the semiclassical approximation. In this case, the physical quantities would undergo higher-order quantum corrections, whose effect on different gravity shares in different forms. Motivated by these facts, in this paper we aim to observe how quantum corrections affect black hole spectroscopy in different gravity. The result shows that, in the presence of higher-order quantum corrections, black hole spectroscopy in different gravity still shares the same form as $S_n=n$, further confirming the entropy quantum is universal in the sense that it is not only independent of black hole parameters, but also independent of higher-order quantum corrections. This is a desiring result for the forthcoming quantum gravity theory.Comment: 14 pages, no figure, use JHEP3.cls. to be published in JHE

Corner contributions to holographic entanglement entropy

The entanglement entropy of three-dimensional conformal field theories contains a universal contribution coming from corners in the entangling surface. We study these contributions in a holographic framework and, in particular, we consider the effects of higher curvature interactions in the bulk gravity theory. We find that for all of our holographic models, the corner contribution is only modified by an overall factor but the functional dependence on the opening angle is not modified by the new gravitational interactions. We also compare the dependence of the corner term on the new gravitational couplings to that for a number of other physical quantities, and we show that the ratio of the corner contribution over the central charge appearing in the two-point function of the stress tensor is a universal function for all of the holographic theories studied here. Comparing this holographic result to the analogous functions for free CFT's, we find fairly good agreement across the full range of the opening angle. However, there is a precise match in the limit where the entangling surface becomes smooth, i.e., the angle approaches $\pi$, and we conjecture the corresponding ratio is a universal constant for all three-dimensional conformal field theories. In this paper, we expand on the holographic calculations in our previous letter arXiv:1505.04804, where this conjecture was first introduced.Comment: 62 pages, 6 figures, 1 table; v2: minor modifications to match published version, typos fixe

Validation of Plasmodium falciparum dUTPase as the target of 5'-tritylated deoxyuridine analogues with anti-malarial activity

BACKGROUND: Malaria remains as a major global problem, being one of the infectious diseases that engender highest mortality across the world. Due to the appearance of resistance and the lack of an effective vaccine, the search of novel anti-malarials is required. Deoxyuridine 5'-triphosphate nucleotido-hydrolase (dUTPase) is responsible for the hydrolysis of dUTP to dUMP within the parasite and has been proposed as an essential step in pyrimidine metabolism by providing dUMP for thymidylate biosynthesis. In this work, efforts to validate dUTPase as a drug target in Plasmodium falciparum are reported. METHODS: To investigate the role of PfdUTPase in cell survival different strategies to generate knockout mutants were used. For validation of PfdUTPase as the intracellular target of four inhibitors of the enzyme, mutants overexpressing PfdUTPase and HsdUTPase were created and the IC50 for each cell line with each compound was determined. The effect of these compounds on dUTP and dTTP levels from P. falciparum was measured using a DNA polymerase assay. Detailed localization studies by indirect immunofluorescence microscopy and live cell imaging were also performed using a cell line overexpressing a Pfdut-GFP fusion protein. RESULTS:Different attempts of disruption of the dut gene of P. falciparum were unsuccessful while a 3' replacement construct could recombine correctly in the locus suggesting that the enzyme is essential. The four 5'-tritylated deoxyuridine analogues described are potent inhibitors of the P. falciparum dUTPase and exhibit antiplasmodial activity. Overexpression of the Plasmodium and human enzymes conferred resistance against selective compounds, providing chemical validation of the target and confirming that indeed dUTPase inhibition is involved in anti-malarial activity. In addition, incubation with these inhibitors was associated with a depletion of the dTTP pool corroborating the central role of dUTPase in dTTP synthesis. PfdUTPase is mainly localized in the cytosol. CONCLUSION: These results strongly confirm the pivotal and essential role of dUTPase in pyrimidine biosynthesis of P. falciparum intraerythrocytic stages