Optical Studies of the coexistence curve of the n-heptane+nitrobenzene mixture near its consolute point measured by an optical method

We have measured the coexistence curve of the binary liquid mixture n-heptane+nitrobenzene near its consolute point using an optical method. In particular, the critical exponent beta describing the coexistence curve was measured for this system. Previous experimental values of beta for n-heptane+nitrobenzene were higher than the typical theoretically calculated value, an unusual, although not unique, occurrence. In an effort to study this discrepancy, we have used an improved experimental apparatus for our measurements. We have taken special care to minimize temperature gradients and maximize the temperature stability of our thermal control system. We have also exploited features of a known optical method to analyze, thoroughly, sources of systematic errors. We measured an apparent value of beta as 0.367+/- 0.006 and by a careful study of the known sources of error we find that they are not able to remove the discrepancy between the measured and the theoretical values of beta. We also measured the critical temperature of the system at Tc=291.80+/- 0.02 K (18.65 C).Comment: 21 pages, 8 figures, submitted to Physical Review B. Shrunk Experimental section, expanded Equilibration and Conclusion sections, eliminated a figure, added reference; 19 pages, 7 figures, resubmitted to PRB. Replaced Fig. 3, added separate simple text file with coexistence curve data (OPvsT.txt); resubmitted to PR

Lorentz-Lorenz Coefficient, Critical Point Constants, and Coexistence Curve of 1,1-Difluoroethylene

We report measurements of the Lorentz-Lorenz coefficient density dependence, the critical temperature, and the critical density, of the fluid 1,1-difluoroethylene. Lorentz-Lorenz coefficient data were obtained by measuring refractive index and density of the same fluid sample independently of one another. Accurate determination of the Lorentz-Lorenz coefficient is necessary for transformation of refractive index data into density data from optics-based experiments on critical phenomena of fluid systems done with different apparatus, with which independent measurement of the refractive indes and density is not possible. Measurements were made along the coexistence curve of the fluid and span the density range 0.01 to 0.80 g/cc. The Lorentz-Lorenz coefficient results show a stronger density dependence along the coexistence curve than previously observed in other fluids, with a monotonic decrease from a density of about 0.2 g/cc onwards, and an overall variation of about 2.5% in the density range studied. No anomaly in the Lorentz-Lorenz coefficient was observed near the critical density. The critical temperature is measured at Tc=(302.964+-0.002) K (29.814 C) and the measured critical density is (0.4195+-0.0018)g/cc.Comment: 14 pages, 6 figures, MikTeX 2.4, submitted to Physical Review

Mechanisms underlying activity of antiretroviral drugs in HIV-1-infected macrophages: New therapeutic strategies

Monocyte-derived macrophages (M/M) are considered the second cellular target of HIV-1 and a crucial virus reservoir. M/M are widely distributed in all tissues and organs, including the CNS, where they represent the most common HIV-infected cells. Differently from activated CD4+ T lymphocytes, M/M are resistant to the cytopathic effect of HIV and survive HIV infection for a long lime. Moreover, HIV-1 replication in M/M is a key pathogenetic event during the course of HIV-1 infection. Overall findings strongly support the clinical relevance of anti-HIV drugs in M/M. Nucleoside RT inhibitors (NRTIs) are more active against HIV in M/M than in CD4+ T lymphocytes. Their activity is further boosted by the presence of an additional monophosphate group (i.e., a phosphonate group, as in the case of Tenofovir), thus overcoming the bottleneck of the low phosphorylation ability of M/M. In contrast, the antiviral activity of non-NRTIs (not affecting the DNA chain elongation) in M/M is similar to that in CD4+ T lymphocytes. Protease inhibitors are the only clinically approved drugs acting at a late stage of the HIV lifecycle. They are able to interfere with HIV replication in HIV-1 chronically infected M/M, even if at concentrations greater than those observed in HIV-1 chronically infected CD4+ T lymphocytes. Finally, several new drugs have been shown to interfere efficiently with HIV replication in M/M, including entry inhibitors. A better understanding of the activity of the anti-HIV drugs in M/M may represent a key element for the design of effective anti-HIV chemotherapy. © Society for Leukocyte Biology

A multi-targeted drug candidate with dual anti-HIV and anti-HSV activity

Human immunodeficiency virus (HIV) infection is often accompanied by infection with other pathogens, in particular herpes simplex virus type 2 (HSV-2). The resulting coinfection is involved in a vicious circle of mutual facilitations. Therefore, an important task is to develop a compound that is highly potent against both viruses to suppress their transmission and replication. Here, we report on the discovery of such a compound, designated PMEO-DAPym. We compared its properties with those of the structurally related and clinically used acyclic nucleoside phosphonates (ANPs) tenofovir and adefovir. We demonstrated the potent anti-HIV and -HSV activity of this drug in a diverse set of clinically relevant in vitro, ex vivo, and in vivo systems including (i) CD4⁺ T-lymphocyte (CEM) cell cultures, (ii) embryonic lung (HEL) cell cultures, (iii) organotypic epithelial raft cultures of primary human keratinocytes (PHKs), (iv) primary human monocyte/macrophage (M/M) cell cultures, (v) human ex vivo lymphoid tissue, and (vi) athymic nude mice. Upon conversion to its diphosphate metabolite, PMEO-DAPym markedly inhibits both HIV-1 reverse transcriptase (RT) and HSV DNA polymerase. However, in striking contrast to tenofovir and adefovir, it also acts as an efficient immunomodulator, inducing β-chemokines in PBMC cultures, in particular the CCR5 agonists MIP-1β, MIP-1α and RANTES but not the CXCR4 agonist SDF-1, without the need to be intracellularly metabolized. Such specific β-chemokine upregulation required new mRNA synthesis. The upregulation of β-chemokines was shown to be associated with a pronounced downmodulation of the HIV-1 coreceptor CCR5 which may result in prevention of HIV entry. PMEO-DAPym belongs conceptually to a new class of efficient multitargeted antivirals for concomitant dual-viral (HSV/HIV) infection therapy through inhibition of virus-specific pathways (i.e. the viral polymerases) and HIV transmission prevention through interference with host pathways (i.e. CCR5 receptor down regulation)

[18F]FDG and [18F]FLT uptake in human breast cancer cells in relation to the effects of chemotherapy: an in vitro study

Increased 2′-deoxy-2′-[18F]fluoro-D-glucose (FDG) uptake is the most commonly used marker for positron emission tomography in oncology. However, a proliferation tracer such as 3′-deoxy-3′-[18F]fluorothymidine (FLT) might be more specific for cancer. 3′-deoxy-3′-[18F]fluorothymidine uptake is dependent on thymidine kinase 1 (TK) activity, but the effects of chemotherapeutic agents are unknown. The aim of this study was to characterise FDG and FLT uptake mechanisms in vitro before and after exposure to chemotherapeutic agents. The effects of 5-fluorouracil (5-FU), doxorubicin and paclitaxel on FDG and FLT uptake were measured in MDA MB231 human breast cancer cells in relation to cell cycle distribution, expression and enzyme activity of TK-1. At IC50 concentrations, 5-FU resulted in accumulation in the G1 phase, but doxorubicin and paclitaxel induced a G2/M accumulation. Compared with untreated cells, 5-FU and doxorubicin increased TK-1 levels by >300. At 72 h, 5-FU decreased FDG uptake by 50% and FLT uptake by 54%, whereas doxorubicin increased FDG and FLT uptake by 71 and 173%, respectively. Paclitaxel increased FDG uptake with >100% after 48 h, whereas FLT uptake hardly changed. In conclusion, various chemotherapeutic agents, commonly used in the treatment of breast cancer, have different effects on the time course of uptake of both FDG and FLT in vitro. This might have implications for interpretation of clinical findings

Evaluación de la degradación de residuos de insecticidas organofosforados en granos de maíz (Zea mays L.) almacenados

The aims of this work were to monitor the  dissipation dynamics of the organophosphate insecticides applied to corn grains stored in  supervised experiments, establish the dissipation dynamics and determine the minimum period post application necessary to comply with the  established commercial standards. The  insecticides dichlorvos (DDVP), hlorpyrifosmethyl+deltamethrin (CPM) and pirimiphos-methyl (PMM) were applied to maize grains, which were then stored for 120 days. The samples were processed and extracted using the QuEChERS technique and  the pesticide residues were identified and  quantified by gas chromatography-mass spectrometry. Deltamethrin residues were not  analized. After 90 days of storage, more than 80 % of the initial residue had been degraded, and after 120 days the residues were below the detection limit (LoD). DDVP, CPM and PMM residues  followed a first order degradation dynamics [Ct = C0×e(-kt)], with k values of 0.384, 0.025 and 0.020 and estimated half-lives of 1.8, 27.7 and 34.7 days, respectively. These results show that at recommended dosages, the insecticides CPM and PMM are safe for use on stored maize grains;  DDVP results provide additional evidence further reasserting its prohibition, which dates back to 2018. Los objetivos del trabajo fueron monitorear la degradación de insecticidas organofosforados aplicados a granos de maíz almacenados en ensayos experimentales, establecer las dinámicas de degradación y determinar el tiempo mínimo de espera post aplicación necesario para cumplir con estándares comerciales. Se aplicaron los insecticidas diclorvós (DDVP), clorpirifós-metil+deltametrina (CPM) y pirimifós-metil (PMM)  a granos de maíz almacenados durante 120 días. Las muestras se procesaron y extrajeron mediante la técnica QuEChERS y los residuos de plaguicidas  se identificaron y cuantificaron mediante  cromatografía de gases-espectrometría de masas.Los residuos de deltametrina no fueron determinados. Luego de 90 días de almacenamiento, más del 80 % del contenido inicial de residuos se había degradado y a los 120  días se encontraban por debajo del límite de detección (LoD). Los residuos de DDVP CPM y PMM siguieron una dinámica de degradación de primer orden [Ct=C0×e(-kt)], con valores de k de 0,384, 0,025 y 0,020 y vidas medias estimadas de 1;8; 27,7 y 34,7 días respectivamente. Estos resultados permiten concluir que a la dosis recomendada, los insecticidas CPM y PMM se pueden usar de manera segura en granos de maíz  almacenados, mientras que los resultados de DDVP aportan información adicional que ratifica su prohibición, que data del 2018

Synthesis, biophysical characterization and anti-HIV activity of d(TG3AG) Quadruplexes bearing hydrophobic tails at the 5'-end

Novel conjugated G-quadruplex-forming d(TG3AG) oligonucleotides, linked to hydrophobic groups through phosphodiester bonds at 50-end, have been synthesized as potential anti-HIV aptamers, via a fully automated, online phosphoramidite-based solid-phase strategy. Conjugated quadruplexes showed pronounced anti-HIV activity with some preference for HIV-1, with inhibitory activity invariably in the low micromolar range. The CD and DSC monitored thermal denaturation studies on the resulting quadruplexes, indicated the insertion of lipophilic residue at the 50-end, conferring always improved stability to the quadruplex complex (20 < DTm < 40 C). The data suggest no direct functional relationship between the thermal stability and anti-HIV activity of the folded conjugated G-quartets. It would appear that the nature of the residue at 50 end of the d(TG3AG) quadruplexes plays an important role in the thermodynamic stabilization but a minor influence on the anti-HIV activity. Moreover, a detailed CD and DSC analyses indicate a monophasic behaviour for sequences I and V, while for ODNs (II–IV) clearly show that these quadruplex structures deviate from simple two-state melting, supporting the hypothesis that intermediate states along the dissociation pathway may exis

Different evolution of genotypic resistance profiles to emtricitabine versus lamivudine in tenofovir-containing regimens.

BACKGROUND: To investigate genotypic resistance profiles to emtricitabine + tenofovir (FTC + TDF) in-vivo and in-vitro, and compare them with lamivudine + tenofovir (3TC + TDF). METHODS: Three hundred fifty-two HIV-1 B-subtype pol sequences from 42 FTC + TDF-treated patients, 40 3TC + TDF-treated patients, and 270 patients treated with 3TC plus another nucleoside reverse transcriptase inhibitor (but not TDF). All patients never received FTC, 3TC, and TDF in their previous therapeutic regimen. 3TC/FTC ± TDF resistance was investigated using in vitro selection experiments and docking simulations. RESULTS: The M184V mutation is less prevalent in FTC + TDF-treated patients than in 3TC + TDF-treated, and 3TC-treated/TDF-naive patients (14.3% versus 40.0%, P = 0.01 and 55.6%, P < 0.001). Multivariable analysis shows that factors correlated with a lower probability of M184V emergence at failure were the use of FTC compared with 3TC [odds ratio (OR): 0.32 (95% confidence interval (CI): 0.10 to 0.99), P = 0.04], the use of boosted protease inhibitor, and the use of TDF [OR: 0.20 (95% CI: 0.11 to 0.37), P < 0.001, and OR: 0.47 (95%CI: 0.22 to 1.01), P = 0.05, respectively]. In vitro selection experiments and docking analysis show that other reverse transcriptase (RT) mutations, even localized in RT connection domain, can be selected by 3TC + TDF or FTC + TDF in M184V absence and can affect RT affinity for 3TC/FTC and/or TDF. CONCLUSIONS: Our study shows lower rates of M184V development in FTC + TDF regimens versus 3TC + TDF and suggests a potential role of boosted protease inhibitors and TDF in delaying the M184V emergence. Novel RT mutational patterns, more complex than currently known, can contribute to 3TC, FTC, and TDF resistance

Surfactant protein D inhibits HIV-1 infection of target cells via interference with gp120-CD4 interaction and modulates pro-inflammatory cytokine production

© 2014 Pandit et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Surfactant Protein SP-D, a member of the collectin family, is a pattern recognition protein, secreted by mucosal epithelial cells and has an important role in innate immunity against various pathogens. In this study, we confirm that native human SP-D and a recombinant fragment of human SP-D (rhSP-D) bind to gp120 of HIV-1 and significantly inhibit viral replication in vitro in a calcium and dose-dependent manner. We show, for the first time, that SP-D and rhSP-D act as potent inhibitors of HIV-1 entry in to target cells and block the interaction between CD4 and gp120 in a dose-dependent manner. The rhSP-D-mediated inhibition of viral replication was examined using three clinical isolates of HIV-1 and three target cells: Jurkat T cells, U937 monocytic cells and PBMCs. HIV-1 induced cytokine storm in the three target cells was significantly suppressed by rhSP-D. Phosphorylation of key kinases p38, Erk1/2 and AKT, which contribute to HIV-1 induced immune activation, was significantly reduced in vitro in the presence of rhSP-D. Notably, anti-HIV-1 activity of rhSP-D was retained in the presence of biological fluids such as cervico-vaginal lavage and seminal plasma. Our study illustrates the multi-faceted role of human SPD against HIV-1 and potential of rhSP-D for immunotherapy to inhibit viral entry and immune activation in acute HIV infection. © 2014 Pandit et al.The work (Project no. 2011-16850) was supported by Medical Innovation Fund of Indian Council of Medical Research, New Delhi, India (www.icmr.nic.in/)