Andrea Geraci

Gender differences in clinical progression of HIV-1-infected individuals during long-term highly active antiretroviral therapy

Objective: To assess gender differences in the long-term clinical, virological and immunological outcomes during highly active antiretroviral therapy (HAART). Methods: This longitudinal observational multicentre study followed 2460 HIV-infected patients who had begun a protease inhibitor-based regimen for a median period of 43 months. Outcome measures were virological suppression (< 500 copies/ml), confirmed virological rebound after suppression, and death or new AIDS-defining illness (ADI). Results: At baseline, 690 female patients (28.0%) had significantly lower age, higher prevalence of heterosexual contact and lower prevalence of intravenous drug use as risk factors for HIV infection compared with males. Furthermore, females had a lower number of AIDS-defining illnesses, higher CD4 cell counts and lower viral loads. No gender differences were reported in terms of proportion of patients achieving viral suppression or exhibiting rebound after achieving viral suppression. Female patients experienced reduced clinical progression during follow-up compared with males (P = 0.008) by Kaplan–Meier analysis; however this difference was not significant in an adjusted analysis. In a multivariate model, the interaction between gender and risk factor for HIV or viral load showed that female drug users and female patients with a baseline HIV RNA viral load of 104–105 copies/ml had a favourable clinical outcome compared with males (P = 0.035 and P = 0.015, respectively). Conclusion: No differences were found between genders in terms of virological and immunological outcomes during long-term HAART. Nevertheless, a lower risk of clinical progression was reported among female patients with intermediate baseline viral load than in males.

Mutagenesis of hepatitis C virus E1 protein affects its membrane-permeabilizing activity

The E1 glycoprotein of hepatitis C virus is a transmembrane glycoprotein with a C-terminal anchor domain. When expressed in Escherichia coli, E1 induces a change in membrane permeability that is toxic to the bacterial cell. The C-terminal hydrophobic region (aa 331-383) of E1 is mainly responsible for membrane association and for inducing changes in membrane permeability. These observed changes are similar to those produced in E. coli by influenza virus M2, human immunodeficiency virus gp41 and poliovirus 3AB proteins, whose hydrophobic domains are thought to cause pore formation in biological membranes. To further characterize the activity of E1 at a molecular level, the membrane-permeabilizing ability of a second internal hydrophobic region (aa 262-291) was examined by expressing different deletion mutants of E1 in an E. coli system that is widely used for analysing membrane-active proteins from other animal viruses. Moreover, highly conserved amino acids in the C-terminal hydrophobic region were mutated to identify residues that are critical for inducing changes in membrane permeability. Analysis of cell growth curves of recombinant cultures and membrane-permeability assays revealed that synthesis of this fragment increased the flux of small compounds through the membrane and caused progressive cell lysis, suggesting that this domain has membrane-active properties. Furthermore, analysis of C-terminal mutants indicated that the conserved amino acids Arg(339), Trp(368) and Lys(370) play a critical role in protein function, as both cell lysis and changes in membrane permeability induced by the wild-type clone could be blocked by substitutions in these positions.

The potential role of garlic (Allium sativum) against the multi-drug resistant tuberculosis pandemic: a review

Worldly data show the increasing incidence of Mycobacterium tuberculosis (MTB) and particularly of multi-drug resistant tuberculosis (MDR-TB). In developing countries, TB control programmes are overwhelmed by the complexity of treating MDR-TB infected people, as current tools and therapies are inadequate. MDR-TB could become the main form of TB. Risk factors that make South Africa into one of the main epicentres are analysed. A review of the studies carried out about antitubercular properties of Allium sativum both in vitro and in vivo is provided. The researches about the garlic extracts effectiveness against clinical isolates of MDR-TB are of scientific importance. Allium sativum offers a hope for developing alternative drugs. The involvement of traditional healers (TH) in the TB health management could facilitate the administration of garlic extracts to the infected patients.

CD4-Independent Infection of Two CD4−/CCR5−/CXCR4+ Pre-T-Cell Lines by Human and Simian Immunodeficiency Viruses

ABSTRACT The infection of CD4-negative cells by variants of tissue culture-adapted human immunodeficiency virus type 1 (HIV-1) or HIV-2 strains has been shown to be mediated by the CXCR4 coreceptor. Here we show that two in vitro-established CD4−/CCR5−/CXCR4+ human pre-T-cell lines (A3 and A5) can be productively infected by wild-type laboratory-adapted T-cell-tropic HIV-1 and HIV-2 strains in a CD4-independent, CXCR4-dependent fashion. Despite the absence of CCR5 expression, A3 and A5 cells were susceptible to infection by the simian immunodeficiency viruses SIVmac239 and SIVmac316. Thus, at least in A3 and A5 cells, one or more of the chemokine receptors can efficiently support the entry of HIV and SIV isolates in the absence of CD4. These findings suggest that to infect cells of different compartments, HIV and SIV could have evolved in vivo to bypass CD4 and to interact directly with an alternative receptor.

Plants and parts of plants used in food supplements: an approach to their safety assessment

In Italy most herbal products are sold as food supplements and are subject only to food law. A list of about 1200 plants authorised for use in food supplements has been compiled by the Italian Ministry of Health. In order to review and possibly improve the Ministrys list an ad hoc working group of Istituto Superiore di Sanità was requested to provide a technical and scientific opinion on plant safety. The listed plants were evaluated on the basis of their use in food, therapeutic activity, human toxicity and in no-alimentary fields. Toxicity was also assessed and plant limitations to use in food supplements were defined.

Clinical researches on the efficacy of spa therapy in fibromyalgia: a systematic review

BACKGROUND Fibromyalgia is characterized by chronic widespread pain, tenderness at muscle and tendon insertions point when digital pressure is applied, sleep disorders, chronic fatigue, depressive episodes, anxiety, and other functional somatic syndromes. OBJECTIVE The aim of this study was to determine whether balneotherapy with mineral waters and mineral-water containing mud is effective in the management of fibromyalgia. METHODS We conducted a systematic review of the literature regarding spa therapy in the treatment of the fibromyalgia. We searched many databases for articles published between 2000 and 2012 and we selected 7 studies among 65 articles retrieved. A total of 142 patients received balneotherapy and 129 were controls. CONCLUSION Study data confirms that spa therapy could improve the symptoms of fibromyalgia including pain, depression and minor symptoms.

Expression and membrane association of hepatitis C virus envelope 1 protein.

The expression of hepatitis C virus (HCV) E1 protein is toxic for Escherichia coli cells. For this reason, we have cloned the E1 gene in the pET3a vector and analyzed the inducible expression of the protein in two strains of E. coli characterised by a different level of reduction of basal synthesis. The results indicated that synthesis of E1 was supported only by the BL21(DE3)pLysS strain which provides a tightest control of protein expression before the induction. The BL21(DE3)pLysS cells were then used for the expression of E1 gene, varying at its carboxy terminus in order to retain (E1, aa 192–383) or delete (E1t, aa 192–340) a C-terminal hydrophobic region that may be involved in membrane association. Following cell fractionation, E1 protein was found associated with the membrane fraction. By contrast, the truncated mutant E1t, was identified in the soluble phase suggesting a direct role for the C-terminal domain in E1 membrane association.

Longitudinal characterization of CD4, CD8 T-cell subsets and of haematological parameters in healthy newborns of cynomolgus monkeys.

A longitudinal characterization of immune cell subpopulations (lymphocytes, CD4+ and CD8+ cells), of routine haematological parameters and of immunoglobulin serum levels was carried out in newborn Macaca fascicularis starting from 1 week up to 1 year of life. In neonates, the percentage of CD4+ lymphocytes is almost double, while the percentage of CD8+ cells is lower than that found in adult monkeys (> 5-years old). An inverted trend in the percentage of the two T-lymphocyte subpopulations was observed during the weeks following birth, with a progressive increase of circulating CD8+, paralleled by a decrease of CD4+ cell number. Consequently, the CD4/CD8 ratio slowly decreases, even if, at 12 months of life, it is still higher than that found in adult animals. Several differences were also noted between young and adult monkeys with regard to the total number of circulating CD4+ and CD8+ cells. Haematological parameters did not show consistent differences with respect to adult values. The plasma IgG level is high at birth, then decreases until 6 months of life, while the IgM and IgA values are very low during the first weeks of life but increase in the following period. Our data showed that variations of immunological (CD4+, CD8+ cells) patterns and of some haematological parameters in M. fascicularis are dependent on age. These variations should be therefore considered whenever young animals are used in experimental protocols.