Involving medical students in re-orienting health services: a photovoice study

Introduction: Healthcare reorientation aims for health services focused not exclusively on diseases but also on prevention and health promotion. The implementation depends strongly on professionals' willingness to actively participate in the reorientation. An effective strategy to boost reorientation is to reorient education and role definition of future professionals. This paper examines whether photovoice can be a suitable method to i) increase future health professionals' awareness of users' needs and expectations; and ii) enable a process of critical reflection on role definition and health services organisation. Methods: One hundred and seventy-two medical students participated in the photovoice project. Participants were asked to produce one photo combined with a caption, responding to a pre-identified question: "What is, in your opinion, the main aspect affecting users' satisfaction/dissatisfaction in a healthcare facility?". Participants discussed their photos in group discussions (n = 16) and participated in data analysis sessions (n = 4). Results: Participants' contributions revolved around how services were delivered (e.g., kindness, accessibility, attention to additional needs) rather than the service provided. The students showed their empathic side and proposed smart and inclusive solutions to improve overall users' experience. Conclusions: This study demonstrated the value of using photovoice to reach medical students and to integrate health promotion into their professional identities. The photovoice process, teamwork, and discussions opened a breach into traditional thinking regarding aspects of healthcare services that are taken for granted or are overlooked. Furthermore, participants' proposals often implied a change in the behaviour of professionals - their future selves - towards patients and low-cost improvements of organisational practices

Kinetic analysis of Human T-cell Leukemia Virus type 2 expression in chronically-infected cells and patient PBMCs

Introduction: The elucidation of the viral gene expression profile provides useful information in assessing the function of specific viral genes in the process of infection and cellular transformation. HTLV-2 pattern of mRNAs expression produces three major classes of mRNAs: unspliced genomic mRNA for Gag, protease and Pol proteins; singly spliced mRNAs encoding Env and the accessory proteins p28, p22/p20-1 and -2; and a doubly spliced mRNA for the regulatory proteins Tax, Rex and for the p10/p11 and p? accessory ones (Ref.1 and Fig. 1). To date, very little information has been obtained on the temporal regulation of different HTLV-2 transcripts expression in infected cells. Aim of this study was to investigate the kinetics of gene expression from HTLV-2 infected cell lines and from PBMCs of HTLV-2B infected subjects. The expression profile and kinetics of the different transcripts were analysed by real time RT-PCR using splice-junction-specific primers. Results: This approach was used to first determine the steady-state levels of expression for the different viral transcripts in three different cell lines in log phase of growth . Experiments performed indicated that gag/pol is the most abundant transcript. The expression level of env was comparable in the two T-cell lines, Mo-T and C344, infected by the 2A subtype, and was considerably higher than in the B-cells infected with HTLV-2B subtype, where p10/p11 and p? transcripts were below the limit of detection. We next investigated the kinetics of viral transcripts expression in infected BJAB-Gu cells. As in the previous experiment, the absolute copy number of gag/pol was the highest over the time period analysed . Among the accessory transcripts, p28,p22/p20-2 was the most abundant while other regulatory and accessory genes were lower. The analysis of fold variation, reported in g. 4B, indicated that tax/rex and p28, p22/p20-1 showed a biphasic profile with an early peak at 24 hours and a second one at 72 hours, whereas the transcripts gag/pol, env and p28,p22/p20-2 were expressed later.The kinetics of gene expression also was analysed from ex-vivo PBMCs of HTLV-2B infected subjects. Fig. 5 shows a typical pattern of expression. Also in this case, among the mRNAs species, gag/pol was consistently the most abundant transcript, p28, p22/p20-2 was approximately 15 fold lower than gag/pol, followed by tax/rex and p? that were present at approximately 25 fold lower than the unspliced mRNA coding for gag/pol . Very low levels of expression were found for p28, p22/p20-1, while env and p10/p11 transcripts were below the limit of detection. In Fig. 5B the fold variation analysis showed that the first mRNAs expressed were tax/rex and p28,p22/p20-1 with a peak at 4 hours followed by all the other transcripts that showed a later peak at 24 hours.These results indicate that tax/rex is the earliest transcript expressed, while the other genes, coding for accessory and structural proteins, are expressed in a later phase of the viral cycle. Conclusions: The expression of different HTLV-2 genes follows a distinct timing both in infected cell lines and PBMCs isolated from infected patients. The transcript tax/rex is the first to be expressed, thus indicating that it is necessary at the beginning of the infection cycle to transactivate and regulate viral and cellular transcripts. These results also suggest that the control of viral gene expression is highly regulated both in its kinetics and expression level

Changes in CD4+ cells’ miRNA expression following exposure to HIV-1

Background: MiRNAs inhibit HIV-1 expression by either modulating host innate immunity or by directly interfering with viral mRNAs. Here, we investigated the miRNA profile that discriminates different classes of HIV-1 infected patients from multiple exposed uninfected individuals. Methods: The expression levels of 377 miRNAs were selectively analyzed in CD4+ cells isolated from whole blood of HIV-1 \ue9lite LTNP (\ue9LTNP), naive, and multiply exposed uninfected individuals (MEU). MiRNA extraction was performed by the mirVana miRNA Isolation Kit (Ambion) and their expression was subsequently examined by real-time PCR-based arrays. The expression of miRNAs was also determined in primary culture of CD4+T cells and monocyte-macrophages infected in vitro by R5 strains. Expression of Dicer and Drosha was evaluated by real-time PCR. Results: We only considered miRNAs that were expressed in the 70% of patients of at least one class and varied by at least 1 log10 from healthy controls. Out of 377 miRNAs, 26 were up-regulated, while 88 were down-regulated. Statistical analysis showed that 21 miRNAs significantly differentiated \ue9LTNP from MEU and 23 miRNAs distinguished naive from MEU, while only 1 (miR-155) discriminated \ue9LTNP from naive. By hierarchical clustering of the miRNAs according to patient class, \ue9LTNP clustered with naive whereas all MEU subjects grouped together. The Dicer and Drosha expression in the patient classes correlated with miRNA profile changes. Among miRNAs differentially expressed in patient classes, 32 were detected in in vitro infection model: the most of the up-regulated miRNAs were expressed in monocyte-macrophages, whereas the most of the down-regulated miRNAs were expressed in T lymphocytes. Conclusions: These findings support that miRNA profile could be the result not only of a productive infection, but also of the exposure to HIV products that leave a signature in immune cells. These data provide some intriguing issues relative to the development of HIV vaccines targeting viral proteins

Analysis of viral transcripts of Human T-cell Leukemia Virus type 2 (HTLV-2) in human T- and B- infected cells and PBMCs from infected subjects

Recent studies carried out on HTLV-1-infected cells have provided information on the relative abundance and timing of expression of different viral transcripts. In the case of HTLV-2, information on the profile and temporal regulation of viral gene expression is still incomplete. To address this point, we set up a splice-junction-specific real-time RT-PCR to evaluate the quantitative level of individual HTLV-2 transcripts. Results obtained led to the quantitation of all the HTLV-2 mRNAs described so far, namely gag/pol, env, tax/rex, p28,p22/p20rex-1, p28,p22/p20rex-2 and p10/p11. This analysis was carried out first on chronically infected lines: T-cells infected with the HTLV-2A subtype (Mo-T and C344) or B-cells infected with the HTLV-2B subtype (BJAB-Gu cells). Results showed different levels of expression of env and tax/rex transcripts in T- and B-cells. The expression profile and kinetics of expression of the different transcripts was analyzed in PBMC obtained ex vivo from infected individuals and cultured for several hours. Current experiments are also aimed at testing the possible connections between the pattern of HTLV-2 gene expression and the different phases of the cell cycle

Delayed progression to AIDS in HTLV-I/HIV-1 coinfected intravenous drug users

Delayed progression to AIDS in HTLV-I/HIV-1 coinfected intravenous drug user