HIV-1 Tat increases the adhesion of monocytes and T-cells to the endothelium in vitro and in vivo: implications for AIDS-associated vasculopathy

HIV-1-infected patients exhibit severe damages of the aortic endothelium, develop angioproliferative lesions such as Kaposi's sarcoma (KS), and have an increased risk of cardiovascular diseases and atherosclerosis. An increased adhesion of leukocytes to the endothelium is a common pathogenic parameter of AIDS-associated vascular diseases. Here we show that the HIV-1 Tat protein, a regulatory protein of HIV-1 released by infected cells, and TNF-alpha, a cytokine increased in sera and tissues of HIV-1-infected patients, activate synergistically the adhesion of leukocytes to endothelial cells both in vitro and in vivo. This effect is selectively mediated by HIV-1 Tat, since HIV-1 Nef, another HIV-1 regulatory protein, and the HIV-1 envelope protein gp41, had no effect. In vitro adhesion assays with PBMC and quantitative cell type analysis of adherent cells by FACS demonstrated that HIV-1 Tat selectively activates the adhesion of T-cells and monocytes but not of B-cells. Intravital microscopic studies in mice confirmed the synergistic activity of HIV-1 Tat and TNF-alpha on leukocyte adhesion to the endothelium in vivo. These data indicate that HIV-1 Tat in cooperation with TNF-alpha may contribute to the vascular damage and cardiovascular diseases observed in AIDS patients but also to the prominent extravasation of T-cells and monocytes which is a key process in the formation and progression of KS lesions.     

Association of BDNF with anorexia, bulimia and age of onset of weight loss in six European populations

Several genes with an essential role in the regulation of eating behavior and body weight are considered candidates involved in the etiology of eating disorders (ED), but no relevant susceptibility genes with a major effect on anorexia nervosa (AN) or bulimia nervosa (BN) have been identified. Brain-derived neurotrophic factor (BDNF) has been implicated in the regulation of food intake and body weight in rodents. We previously reported a strong association of the Met66 allele of the Val66Met BDNF variant with restricting AN (ANR) and low minimum body mass index in Spanish patients. Another single nucleotide polymorphism located in the promoter region of the BDNF gene (-270C>T) showed lack of association with any ED phenotype. In order to replicate these findings in a larger sample, we performed a case-control study in 1142 Caucasian patients with ED consecutively recruited in six different centers from five European countries (France, Germany, Italy, Spain and UK) participating in the 'Factors in Healthy Eating' project. We have found that the Met66 variant is strongly associated to all ED subtypes (AN, ANR, binge-eating/purging AN and BN), and that the -270C BDNF variant has an effect on BN and late age at onset of weight loss. These are the first two variants associated with the pathophysiology of ED in different populations and support a role for BDNF in the susceptibility to aberrant eating behaviors.     

Grip force control during gait initiation with a hand-held object

When walking with a hand-held object, grip force is coupled in an anticipatory manner to changes in inertial force resulting from the accelerations and decelerations of gait. However, it is not known how grip and inertial forces are organized at the onset of gait, and if the two forces are coupled in the early phases of gait initiation. Moreover, initiating walking with an object involves the coordination of anticipatory postural (e.g., ground reaction force changes) and grasping adjustments. The aim of this study was to investigate the relationship of ground reaction, grip, and inertial force onsets, and the subsequent development of the coupling of grip and inertial forces during gait initiation with a hand-held object. Ten subjects performed gait initiation with a hand-held object following predictable and unpredictable start signals. We found that ground reaction and grip force onsets were closely linked in time regardless of the predictability of the start signal. In the early period of gait initiation, the grip force started to increase prior to inertial force changes. While the strength of the coupling of grip and inertial forces was moderate in this early phase, it increased to values observed during steady-state gait after the swing foot left the ground. The early grip force increase and the coupling of grip and inertial forces represent an anticipatory control process. This process establishes an appropriate grip-inertial force ratio to ensure object stability during acceleration after foot-off and maintains this increased ratio thereafter. The results suggest that grasping and whole body movements are governed by a common internal representation.     

Mesenchymal stem cell inhibition of T-helper 17 cell- differentiation is triggered by cell-cell contact and mediated by prostaglandin E2 via the EP4 receptor

Mesenchymal stem cells (MSCs) inhibit T‐cell activation and proliferation but their effects on individual T‐cell‐effector pathways and on memory versus naïve T cells remain unclear. MSC influence on the differentiation of naïve and memory CD4⁺ T cells toward the Th17 phenotype was examined. CD4⁺ T cells exposed to Th17‐skewing conditions exhibited reduced CD25 and IL‐17A expression following MSC co‐culture. Inhibition of IL‐17A production persisted upon re‐stimulation in the absence of MSCs. These effects were attenuated when cell–cell contact was prevented. Th17 cultures from highly purified naïve‐ and memory‐phenotype responders were similarly inhibited. Th17 inhibition by MSCs was reversed by indomethacin and a selective COX‐2 inhibitor. Media from MSC/Th17 co‐cultures contained increased prostaglandin E2 (PGE2) levels and potently suppressed Th17 differentiation in fresh cultures. MSC‐mediated Th17 inhibition was reversed by a selective EP4 antagonist and was mimicked by synthetic PGE2 and a selective EP4 agonist. Activation‐induced IL‐17A secretion by naturally occurring, effector‐memory Th17 cells from a urinary obstruction model was also inhibited by MSC co‐culture in a COX‐dependent manner. Overall, MSCs potently inhibit Th17 differentiation from naïve and memory T‐cell precursors and inhibit naturally‐occurring Th17 cells derived from a site of inflammation. Suppression entails cell‐contact‐dependent COX‐2 induction resulting in direct Th17 inhibition by PGE2 via EP4.     

The L76V mutation in HIV-1 protease is potentially associated with hypersusceptibility to protease inhibitors Atazanavir and Saquinavir: is there a clinical advantage?

Background

We aimed to determine the reliability and validity of a hepatitis symptom inventory and to identify predictors of hepatitis C (HCV) treatment initiation in a cohort of HIV-infected patients.

Methods

Prospective clinic based study that enrolled patients referred for HCV therapy consideration. A hepatitis symptom inventory and the Center for Epidemiologic Studies Depression Scale (CES-D) were administered to HIV/HCV individuals. The symptom inventory was factor analyzed and subscale reliability estimated with Cronbach's alpha. Predictive validity was evaluated using generalized estimating equations (GEE). Predictors of HCV treatment were identified using logistic regression.

Results

Between April 2008 to July 2010, 126 HIV/HCV co-infected patients were enrolled in the study. Factor analysis using data from 126 patients yielded a three-factor structure explaining 60% of the variance for the inventory. Factor 1 (neuropsychiatric symptoms) had 14 items, factor 2 (somatic symptoms) had eleven items, and factor 3 (sleep symptoms) had two items, explaining 28%, 22% and 11% of the variance, respectively. The three factor subscales demonstrated high intrinsic consistency reliability. GEE modeling of the 32 patients who initiated HCV therapy showed that patients developed worsening neuropsychiatric and somatic symptoms following HCV therapy with stable sleep symptoms. Bivariate analyses identified the following as predictors of HCV therapy initiation: lower HIV log₁₀ RNA, lower scores for neuropsychiatric, somatic and sleep symptoms, lower CES-D scores and white ethnicity. In stepwise multiple logistic regression analysis, low neuropsychiatric symptom score was the strongest independent predictor of HCV therapy initiation and HIV log₁₀ RNA was inversely associated with a decision to initiate HCV treatment.

Conclusions

A 41-item hepatitis-related symptom inventory was found to have a clinically meaningful 3-factor structure with excellent internal consistency reliability and predictive validity. In adjusted analysis, low neuropsychiatric symptom scores and controlled HIV infection were independent predictors of HCV treatment initiation. The usefulness of the HCV symptom inventory in monitoring HCV treatment should be evaluated prospectively.     

A Novel Bifunctional Trithiocarbonate for Styrene and Methacrylate RAFT Polymerization

The synthesis of butane‐1,4‐bis(phenylacetonitrile trithiocarbonate) and its application as a new chain transfer agent for controlled radical polymerization is reported. It is a bifunctional trithiocarbonate for the polymerization of methacrylates, and styrenic monomers and therefore a novel type of RAFT agent. Linear growth of molar masses with conversion and narrow molecular weight distributions are obtained. ABA triblock copolymers are successfully synthesized in two steps. MALDI‐TOF MS and SEC prove the living character of the polymers and uniform growth of both functionalities. The pre‐equilibrium kinetics are analyzed by SEC to determine transfer coefficients. The P(GMA) blocks are used as precursor for polymer‐analogous reactions.

     

N‐oxyl mediated free radical donor‐acceptor co‐ and terpolymerization of styrene,cyclic maleimide monomers and n‐butyl methacrylate

The N‐oxyl mediated donor‐acceptor copolymerization of styrene as a donor and the maleimides N‐phenylmaleimide (NPI), N‐benzylmaleimide (BMI), and N‐cyclohexylmaleimide (CMI) as acceptor monomers is studied, using 2,2,6,6‐tetramethylpiperidine‐N‐oxyl (TEMPO) as reversible capping agent and benzoyl peroxide (BPO) as initiator. These copolymerizations are characterized by high polymerization rates and very short induction periods. In bulk, a control of the molecular weights and the polydispersity is only possible for the weakest acceptor CMI. For the stronger acceptors BMI and NPI the polymerizations have to be performed in solution (anisol) to realize a linear increase of the molecular weight with conversion and polydispersities below M_w/M_n 1.55. The polymerization rates (NPI > BMI > CMI), the induction periods (CMI > BMI > NPI) and the total number of polymer chains (NPI > BMI > CMI) of the TEMPO mediated free radical donor‐acceptor copolymerizations are a function of the acceptor strength of the maleimide monomers. As the main reason for the observed high reactivity, an “acceptor depending self‐initiation”, proceeding via a Diels‐Alder reaction mechanism, is discussed. Additionally, the concept of the “acceptor depending self‐initiation” is used for the rate of acceleration of S/BuMA copolymerizations leading to poly(S‐co‐BuMA‐co‐maleimide) terpolymers with well‐controlled molecular weights and narrow molecular weight distributions.     

Transparent, elastomeric and tough hydrogels from poly(ethylene glycol) and silicate nanoparticles

The structures and mechanical properties of both physically and covalently cross-linked nanocomposite hydrogels made from poly(ethylene glycol) (PEG) and silicate nanoparticles (Laponite RD) are investigated. Injectable nanocomposite precursor solutions can be covalently cross-linked via photopolymerization. The resulting hydrogels are transparent and have interconnected pores, high elongation and toughness. These properties depend on the hydrogel composition, polymer-nanoparticle interactions and degree of cross-linking (both physical and covalent). Covalent cross-linking of polymer chains leads to the formation of an elastic network, whereas physical cross-linking between nanoparticles and polymer chains induces viscoelastic properties. At high deformations covalent bonds may be broken but physical bonds rebuild and to some extent self-heal the overall network structure. Addition of silicate also enhances the bioactivity and adhesiveness of the hydrogel as these materials stick to soft tissue as well as to hard surfaces. In addition, MC3T3-E1 mouse preosteoblast cells readily adhere and spread on nanocomposite hydrogel surfaces. Collectively, the combinations of properties such as elasticity, stiffness, interconnected network, adhesiveness to surfaces and bio-adhesion to cells provide inspiration and opportunities to engineer mechanically strong and elastic tissue matrixes for orthopedic, craniofacial and dental applications.