Human immunodeficiency virus type 1 (HIV-1) protease inhibitors block cell-to-cell HIV-1 endocytosis in dendritic cells

Sexual transmission is now the most frequent means of diffusion of human immunodeficiency virus type 1 (HIV-1). Even if the underlying mechanism is still largely unknown, there is a consensus regarding the key role played by mucosal dendritic cells (DCs) in capturing HIV through contact with infected subepithelial lymphocytes, and their capacity to spread HIV by trans-infection. We found that HIV protease inhibitors (PIs) reduced virion endocytosis strongly in monocyte-derived immature (i) DCs contacting HIV-1-infected cells, and that this phenomenon led to dramatically impaired trans-infection activity. This inhibitory effect was not mediated by the block of viral protease activity, as it was also operative when donor cells were infected with a PI-resistant HIV-1 strain. The block of virus maturation imposed by PIs did not correlate with significant variations in the levels of virus expression in donor cells or of Gag/Env virion incorporation. Also, PIs did not affect the endocytosis activity of DCs. In contrast, we noticed that PI treatment inhibited the formation of cell–cell conjugates whilst reducing the expression of ICAM-1 in target iDCs. Our results contribute to a better delineation of the mechanisms underlying HIV-1 trans-infection activity in DCs, whilst having implications for the development of new anti-HIV microbicide strategies

Em busca de um ethos siciliano

Using Mainguenau’s concept of ethos, this article aims to outline, by analyzing the literary discourse of some Sicilian writers, which was accepted, repeated and passed on by the own inhabitants of the island, the various ethé linked to the Mediterranean island. This starts at the point in which the clarity that a typical ‘identity’ and ‘character’ is nothing more than a speech or, rather, speeches, in which such characteristics, according to the period and author, came to have different names.Servindo-se do conceito de ethos de Mainguenau, o presente artigo procura delinear, mediante a análise do discurso literário de alguns escritores sicilianos, a seguir aceito, reiterado e retransmitido pelos próprios habitantes da ilha, os diversos ethé ligados à ilha mediterrânea, partindo da clareza de que uma ‘identidade’ e um ‘caráter’ típicos nada mais são do que um discurso, ou melhor, discursos, em que tais características, conforme a época e o autor, vieram a ter denominações diferentes

Lower cost of transport coincides with self-selected walking speed in pregnant individuals

Pregnancy has important structural and physiological changes related to the growth of the baby [1]. These changes can alter metabolic and biomechanical parameters of pregnant individuals (PI) [2]. Changes in the body centre of mass (CM), body dimensions, and weight can influence postural aspects as well as gait [3]. Thus, changes may occur in different variables such as cost of transport (C J.kg-1.m-1) [4], ventilatory efficiency evaluated by the ventilatory equivalent for carbon dioxide (VE/VCO2), and walking speed. Aim: to compare C, VE/VCO2, self-selected walking speed (SSWS) on treadmill and on a track, at different walking speeds, of PI and a control group (CG). PI: n=24 (n=8 in 1st trimester (T1), 12-13 wks; n=8 in 2nd trim. (T2), 26-27 wks; n=8 in 3rd trim. (T3), 35-36 wks); CG: n=8 healthy non-PI. VO2 uptake was collected during trials of 5 min at 5 walking speeds, in randomized order (SSWS, ±40%, ±20% of SSWS) on the treadmill [5]. The SSWS was initially assessed on a track, and then tested on the treadmill. ANOVA for repeated measures was used to compare C and VE/VCO2 for walking speed and groups. One-way ANOVA was used to compare SSWS on the track and on the treadmill. Multiple comparisons were possibly made with Tukey’s post-hoc analysis (α=0.05). Values of C were higher in PI compared to CG for all T (p=0.005; differences between trimesters p=0.001). VE/VCO2 also showed higher values for PI (among T p<0.0001) compared to CG (p<0.0001). Differences in the SSWS (T1 0.83±0.14; T2 0.82±0.18; T3 0.85±0.18; CG 1.01±0.14 m.s-1) on the treadmill and on the track were found for all T (p<0.0001), and between PI and CG (p<0.0001). C always showed the characteristics of "U" shape described previously [4], with higher metabolic economy at SSWS. Interestingly, VE/VCO2 showed higher values in T2 compared to T1, T3 and CG. VE/VCO2 is known to be affected by increased chemoreceptor numbers, peripheral ergoreceptor response, dead space ventilation, and muscle mass involved in exercise [2]. The feeling of greater respiratory difficulty with the increase in walking speed and due to the smaller excursion of the diaphragm by the increased abdominal volume. Our results showed that the greatest respiratory comfort (lower VE/VCO2) occurred at the highest speed performed (above SSWS). The choice of SSWS seems to be related to a greater metabolic economy and not to respiratory comfort. After all physiological changes in T1 and T2 of pregnancy [1], T3 resulted to a certain adaptation. Lower metabolic economy is probably due to the physical alterations such as increase in body mass, displacement of the CM which could have altered the pendulum mechanism, generating displacement with less mechanical efficiency.Agencia Nacional de Investigación e Innovació

The impact of national and international guidelines on newborn care in the nurseries of Piedmont and Aosta Valley, Italy

BACKGROUND: Care procedures for preventing neonatal diseases are carried out according to nurseries' traditions and may be not consistent with the evidence based medicine issues. METHODS: A multi-centric survey was conducted in 2 Regions located in NW Italy (Piedmont and Aosta Valley) in order to collect information on some healthy newborn care procedures. During 2001, a questionnaire was sent to the chief pediatrician in charge to the all 33 nurseries of the region asking the methods used during 2000 as prevention of ophthalmia neonatorum, early and late hemorrhagic disease of newborn, umbilical cord care and recommendations of vitamin D administration. Thereafter, during 2004 the same questionnaire was sent to the 34 chief pediatrician of nurseries to evaluate if the procedures were changed during 2003 according to guidelines. The nurseries care for 32,516 newborns in 2000 and 37,414 in 2003. RESULTS: Aminoglycoside eyes drops as prevention of ophthalmia neonatorum were the first choice in both periods (23 out 33 nurseries in 2000 and 24 out 34 in 2003 p > 0.05; the corresponding figures for newborns were18,984 out 32,516 newborns vs. 28,180 out of 37,414 p < 0.05). The umbilical cord care was carried out with alcohol in 12/33 centers (13,248 newborns) and dry gauze in 3/33 centers (2,130 newborns) in 2000, the corresponding figures in 2003 were 6/34 centers (p > 0.05), (6,380 newborns, p < 0.05) and 12/34 centers (p < 0.05), (18,123 newborns, p < 0.05). The percentage of newborns receiving of i.m. vitamin K. at birth increased during the study period (15,923/32,104 in 2000 vs. 19,684/37,414 in 2003, p < 0.01), but not the number of nurseries (16 in 2000 and 17 in 2003 p > 0.05). The numbers of parents of newborns who receive the recommendations of oral vitamin K during the first months life decreased from 2000 (25,516/30,606) to 2003 (29,808/37,414, p < 0.01) as well as for Vitamin D recommendation (14,582/30,616 in 2000 vs. 11,051/37,414 in 2003, p < 0.01). Oral vitamin K during the first months of life was recommended by 25 nurseries in 2000 and 27 in 2003 (p > 0.05), the corresponding figures for Vitamin D were 15 and 14 (p > 0.05). CONCLUSION: In the present study a large variability of procedures among the nurseries was observed. During the study periods, guidelines and evidence based medicine issues have only partially modified the neonatal care procedures In Piedmont and Aosta Valley nurseries. These observations suggest to implement local forum/consensus conference to standardized procedures as much as possible

Endogenous CCL2 neutralization restricts HIV-1 replication in primary human macrophages by inhibiting viral DNA accumulation

Macrophages are key targets of HIV-1 infection. We have previously described that the expressionof CC chemokine ligand 2 (CCL2) increases during monocyte differentiation to macrophages and it is furtherup-modulated by HIV-1 exposure. Moreover, CCL2 acts as an autocrine factor that promotes viral replication ininfected macrophages. In this study, we dissected the molecular mechanisms by which CCL2 neutralization inhibitsHIV-1 replication in monocyte-derived macrophages (MDM), and the potential involvement of the innate restrictionfactors protein sterile alpha motif (SAM) histidine/aspartic acid (HD) domain containing 1 (SAMHD1) and apolipoproteinB mRNA-editing, enzyme-catalytic, polypeptide-like 3 (APOBEC3) family members.Results:CCL2 neutralization potently reduced the number of p24 Gag+cells during the course of either productive orsingle cycle infection with HIV-1. In contrast, CCL2 blocking did not modify entry of HIV-1 based Virus Like Particles, thusdemonstrating that the restriction involves post-entry steps of the viral life cycle. Notably, the accumulation of viralDNA, both total, integrated and 2-LTR circles, was strongly impaired by neutralization of CCL2. Looking for correlates ofHIV-1 DNA accumulation inhibition, we found that the antiviral effect of CCL2 neutralization was independent of themodulation of SAMHD1 expression or function. Conversely, a strong and selective induction of APOBEC3A expression,to levels comparable to those of freshly isolated monocytes, was associated with the inhibition of HIV-1 replicationmediated by CCL2 blocking. Interestingly, the CCL2 neutralization mediated increase of APOBEC3A expression was typeI IFN independent. Moreover, the transcriptome analysis of the effect of CCL2 blocking on global gene expressionrevealed that the neutralization of this chemokine resulted in the upmodulation of additional genes involved in thedefence response to viruses.Conclusions:Neutralization of endogenous CCL2 determines a profound restriction of HIV-1 replication in primaryMDM affecting post-entry steps of the viral life cycle with a mechanism independent of SAMHD1. In addition, CCL2blocking is associated with induction of APOBEC3A expression, thus unravelling a novel mechanism which mightcontribute to regulate the expression of innate intracellular viral antagonistsin vivo. Thus, our study may potentially leadto the development of new therapeutic strategies for enhancing innate cellular defences against HIV-1 and protecting macrophages from infection

Nonintegrating Lentiviral Vector-Based Vaccine Efficiently Induces Functional and Persistent CD8+ T Cell Responses in Mice

CD8+ T cells are an essential component of an effective host immune response to tumors and viral infections. Genetic immunization is particularly suitable for inducing CTL responses, because the encoded proteins enter the MHC class I processing pathway through either transgene expression or cross-presentation. In order to compare the efficiency and persistence of immune response induced by genetic vaccines, BALB/c mice were immunized either twice intramuscularly with DNA plasmid expressing a codon-optimized HIV-1 gp120 Envelope sequence together with murine GM-CSF sequence or with a single immunization using an integrase defective lentiviral vector (IDLV) expressing the same proteins. Results strongly indicated that the schedule based on IDLV vaccine was more efficient in inducing specific immune response, as evaluated three months after the last immunization by IFNγ ELISPOT in both splenocytes and bone marrow- (BM-) derived cells, chromium release assay in splenocytes, and antibody detection in sera. In addition, IDLV immunization induced high frequency of polyfunctional CD8+ T cells able to simultaneously produce IFNγ, TNFα, and IL2

Successful Immunization with a Single Injection of Non-integrating Lentiviral Vector

We evaluated the ability of an integrase (IN)-defective self-inactivating lentiviral vector (sinLV) for the delivery of human immunodeficiency virus-1 (HIV-1) envelope sequences in mice to elicit specific immune responses. BALB/c mice were immunized with a single intramuscular injection of the IN-defective sinLV expressing the codon optimized HIV-1 JR-FL gp120 sequence, and results were compared with those for the IN-competent counterpart. The IN-defective sinLV elicited specific and long-lasting immune responses, as evaluated up to 90 days from the immunization by enzyme-linked immunosorbent spot (ELISPOT) and intracellular staining (ICS) for interferon- γ (IFN- γ ) assays in both splenocytes and bone marrow (BM) cells, chromium release assay in splenocytes, and antibody detection in sera, without integration of the vector into the host genome. These data provide evidence that a single administration of an IN-defective sinLV elicits a significant immune response in the absence of vector integration and may be a safe and useful strategy for vaccine development