- [Show abstract] [Hide abstract] ABSTRACT: Pneumocystis jirovecii causes severe interstitial pneumonia (PcP) in immunosuppressed patients. This multicenter study assessed the distribution frequencies of epidemiologically relevant genetic markers of P. jirovecii in different geographic populations from Portugal, the USA, Spain, Cuba and Mozambique, and the relationship between the molecular data and the geographical and clinical information, based on a multifactorial approach. The high-throughput typing strategy for P. jirovecii characterization consisted of DNA pooling using quantitative real-time PCR (qPCR) followed by multiplex-PCR/single base extension (MPCR/SBE). The frequencies of relevant P. jirovecii single nucleotide polymorphisms (SNP) (mt85, SOD110, SOD215, DHFR312, DHPS165 and DHPS171) encoded at four loci were estimated in 10 DNA pooled samples representing a total of 182 individual samples. Putative multilocus genotypes (MLG) of P. jirovecii showed to be clustered due to the geographic differences but also dependent on clinical characteristics of the populations studied. The haplotype DHFR312T/SOD110C/SOD215T was associated with severe AIDS-related PcP and high P. jirovecii burdens. The frequencies of this genetic variant of P. jirovecii were significantly higher in patients with AIDS-related PcP from Portugal and the USA than in the colonized patients from Portugal, Spain, Cuba or Mozambique, highlighting the importance of this haplotype, apparently associated with the severity of the disease. Patients from the USA and Mozambique showedhigher rates of DHPS mutants, which may suggest the circulation of P. jirovecii organisms potentially related with TMP-SMX resistance in those geographical regions. This report assessed the worldwide distribution of P. jirovecii haplotypes and their epidemiological impact in distinct geographic and clinical populations.
- [Show abstract] [Hide abstract] ABSTRACT: Single-dose nevirapine (sd-NVP) has been the main option for prevention of mother-to-child transmission (PMTCT) of HIV-1 in low-resource settings. However, sd-NVP can induce the selection of HIV-1 resistant mutations in mothers and infants. In Mozambique, there are limited data regarding the profile of NVP resistance associated mutations (RAM) in the context of PMTCT. To assess the prevalence and the factors associated with NVP RAM among children born to HIV-1 infected mothers enrolled in the PMTCT programme adopted in Mozambique. One hundred and fifty seven children aged 6 to 48 weeks were sequentially included (July 2011 to March 2012) at four centres in Maputo. Genotyping of RAM was performed in samples with HIV-1 RNA≥ 100 copies/μL (Viroseq). Sequencing was performed with ABI 3100 (Applied Biosystems). Logistic regression modelling was undertaken to identify the factors associated with NVP RAM. Seventy-nine children had their samples genotyped. Their median age was 7.0 (3-12) months and 92.4% received prophylaxis with sd-NVP at birth plus daily NVP. 35.4% of mothers received antiretrovirals (ARVs) for PMTCT. ARV RAM were detected in 43 (54.4%) of the children. 45.6% of these children had at least one NVP RAM. The most common mutations associated with NVP resistance were K103N (n = 16) and Y181C (n = 15). NVP RAM was significantly associated with mother exposure to PMTCT (crude odds ratio [OR] 30.3, 95% CI 4.93-186.34) and with mother's CD4 count < 350 cells/mm3 (crude OR 3.08, 95% CI 1.02-9.32). In the multivariable analysis the mother's exposure to PMTCT was the only variable significantly associated with NVP RAM (adjusted OR 48.65, 95% CI 9.33-253.66). We found a high prevalence of NVP RAM among children who were exposed to the drug regimen for PMTCT in Mozambique. The mothers' exposure to PMTCT significantly increased the risk of NVP RAM.
- [Show abstract] [Hide abstract] ABSTRACT: Chronic kidney disease (CKD) is a major health issue for HIV-positive individuals, associated with increased morbidity and mortality. Development and implementation of a risk score model for CKD would allow comparison of the risks and benefits of adding potentially nephrotoxic antiretrovirals to a treatment regimen and would identify those at greatest risk of CKD. The aims of this study were to develop a simple, externally validated, and widely applicable long-term risk score model for CKD in HIV-positive individuals that can guide decision making in clinical practice.
- [Show abstract] [Hide abstract] ABSTRACT: Cobicistat (COBI) is a pharmacoenhancer with no antiretroviral activity. International, randomized double-blind active-controlled trial to evaluate the efficacy and safety of COBI vs ritonavir (RTV) as a pharmacoenhancer of atazanavir in combination with emtricitabine/tenofovir disoproxil fumarate in HIV treatment-naive patients followed through week 144. At Week 144, virologic suppression was achieved in 72% (COBI) and 74% (RTV) of patients. Adverse events leading to study drug discontinuation occurred in 11% of patients in each group. Median changes in serum creatinine (mg/dL) were +0.13 (COBI) and +0.07 (RTV) and were unchanged from week 48. Once-daily COBI is a safe and effective pharmacoenhancer of the protease inhibitor atazanavir.
- [Show abstract] [Hide abstract] ABSTRACT: The diagnosis of Pneumocystis pneumonia (PcP) relies on microscopic visualization of P. jirovecii organisms or DNA detection in pulmonary specimens. This study aimed to assess the usefulness of (1-3)-β-D-Glucan (BG), Krebs von den Lungen-6 antigen (KL-6), lactate dehydrogenase (LDH) and S-adenosyl Methionine (SAM) as serological biomarkers in the diagnosis of PcP. Serum levels of BG, KL-6, LDH and SAM were investigated in 145 Portuguese patients, 50 patients from The Netherlands, 25 Spanish patients and 40 Portuguese blood donors. Data on clinical presentation, chest imaging and gasometry tests were available. PcP cases were confirmed by microscopy and PCR techniques. A cost-effectiveness analysis was performed. BG showed to be the most reliable serologic biomarker for PcP diagnosis, followed by KL-6, LDH and SAM. The combination test BG/KL-6 demonstrated to be the most accurate serologic approach for PcP diagnosis with 94.3% sensitivity and 89.6% specificity. Although less sensitive/specific than the gold standard classic methods based on bronchoalveolar lavage followed by microscopic or molecular detection of P. jirovecii organisms, the BG/KL-6 test may provide a less onerous procedure for PcP diagnosis, using a minimally invasive and inexpensive specimen (blood), which may be also a major benefit for the patient’s care. The BG/KL-6 test should be interpreted within the clinical context and it may be used as a preliminary screening test in patients with primary suspicion of PcP, or as an alternative diagnostic procedure in patients with respiratory failure, or in children, avoiding associated risk of complications by the use of bronchoscopy.
- [Show abstract] [Hide abstract] ABSTRACT: Objective: To assess the relationship between platelet counts and risk of AIDS and non-AIDS-defining events. Design: Prospective cohort. Methods: EuroSIDA patients with at least one platelet count were followed from baseline (first platelet ≥ 1 January 2005) until last visit or death. Multivariate Poisson regression was used to assess the relationship between current platelet counts and the incidence of non-AIDS-defining (pancreatitis, end-stage liver/renal disease, cancer, cardiovascular disease) and AIDS-defining events. Results: There were 62 898 person-years of follow-up (PYFU) among 12 279 patients, including 1168 non-AIDS-defining events [crude incidence 18.6/1000 PYFU, 95% confidence interval (CI) 17.5–19.6] and 735 AIDS-defining events (crude incidence 11.7/1000 PYFU, 95% CI 10.8–12.5). Patients with thrombocytopenia (platelet count ≤100 × 109/l) had a slightly increased incidence of AIDS-defining events [adjusted incidence rate ratio (aIRR) 1.42, 95% CI 1.07–1.86], when compared to those with platelet counts 101–200 × 109/l, whereas the incidence of non-AIDS-defining events was more than two-fold higher (aIRR 2.66, 95% CI 2.17–3.26). Among non-AIDS-defining events, the adjusted incidence of cancer (aIRR 2.20, 95% CI 1.61–3.01), but not cardiovascular disease (aIRR 0.66, 95% CI 0.32–1.34), was significantly higher in patients with thrombocytopenia. The association between thrombocytopenia and cancer remained unaltered in sensitivity analyses requiring repeated platelet counts to confirm thrombocytopenia and lagging platelets by 1 year prior to clinical events. Conclusion: Patients with thrombocytopenia had increased incidence of AIDS-defining and non-AIDS-defining events, but the association with the latter, in particular cancer, was stronger. Future studies should investigate whether the pathophysiological processes underlying thrombocytopenia are associated with the development of cancer during treated HIV disease.
- [Show abstract] [Hide abstract] ABSTRACT: Rare systemic studies concerning prevalence of intestinal parasites in children have been conducted in the second smallest country in Africa, the Democratic Republic of São Tomé and Príncipe. Fecal specimens from 348 children (214 in-hospital attending the Aires de Menezes Hospital and 134 from Agostinho Neto village) in São Tome Island were studied by parasitological and molecular methods. Of the 134 children from Agostinho Neto, 52.2% presented intestinal parasites. 32.1% and 20.2% of these children had monoparasitism and polyparasitism, respectively. Ascaris lumbricoides (27.6%), G. duodenalis (7.5%), T. trichiura (4.5%) and Entamoeba coli (10.5%) were the more frequent species identified in the children of this village. Giardia duodenalis (7.5%) and E. bieneusi (5.2%) were identified by PCR. Nested-PCR targeting G. duodenalis TPI identified Assemblage A (60%) and Assemblage B (40%). The E. bieneusi ITS-based sequence identified genotypes K (57.1%), KIN1 (28.6%) and KIN3 (14.3%). Among the 214 in-hospital children, 29.4% presented intestinal parasites. In 22.4% and 7.0% of the parasitized children, respectively, one or more species were concurrently detected. By microscopy, A. lumbricoides (10.3%) and Trichiuris trichiura (6.5%) were the most prevalent species among these children, and Cryptosporidium was detected by PCR in 8.9% of children. GP60 locus analysis identified 6.5% of C. hominis (subtypes IaA27R3 [35.7%], IaA23R3 [14.3%], IeA11G3T3 [28.6%] and IeA11G3T3R1 [21.4%]) and 2.3% of C. parvum (subtypes IIaA16G2R1 [20.0%], IIaA15G2R1 [20.0%], IIdA26G1 [40.0%] and IIdA21G1a [20.0%]). G. duodenalis and E. bieneusi were identified in 0.5% and 8.9% of the in-hospital children, respectively. G. duodenalis Assemblage B was characterized. The E. bieneusi genotypes K (52.6%), D (26.4%), A (10.5%) and KIN1 (10.5%) were identified. Although further studies are required to clarify the epidemiology of these infectious diseases in this endemic region the significance of the present results highlights that it is crucial to strength surveillance on intestinal pathogens.
- [Show abstract] [Hide abstract] ABSTRACT: Pneumocystis pneumonia (PcP) is a major HIV-related illness caused by Pneumocystis jirovecii. Definitive diagnosis of PcP requires microscopic detection of P. jirovecii in pulmonary specimens. The objective of this study was to evaluate the usefulness of two serum markers in the diagnosis of PcP. Serum levels of (1-3)-beta-d-glucan (BG) and lactate dehydrogenase (LDH) were investigated in 100 HIV-positive adult patients and 50 healthy blood donors. PcP cases were confirmed using indirect immunofluorescence with monoclonal anti-Pneumocystis antibodies and nested-PCR to amplify the large subunit mitochondrial rRNA gene of P. jirovecii in pulmonary specimens. BG and LDH levels in serum were measured using quantitative microplate-based assays. BG and LDH positive sera were statistically associated with PcP cases (P ≤ 0.001). Sensitivity, specificity, positive/negative predictive values (PPV/NPV), and positive/negative likelihood ratios (PLR/NLR) were 91.3 %, 61.3 %, 85.1 %, 79.2 %, 2.359, and 0.142, respectively, for the BG kit assay, and 91.3 %, 35.5 %, 75.9 %, 64.7 %, 1.415 and 0.245, respectively, for the LDH test. Serologic markers levels combined with the clinical diagnostic criteria for PcP were evaluated for their usefulness in diagnosis of PcP. The most promising cutoff levels for diagnosis of PcP were determined to be 400 pg/ml of BG and 350 U/l of LDH, which combined with clinical data presented 92.8 % sensitivity, 83.9 % specificity, 92.8 % PPV, 83.9 % NPV, 5.764 PLR and 0.086 NLR (P < 0.001). This study confirmed that BG is a reliable indicator for detecting P. jirovecii infection. The combination between BG/LDH levels and clinical data is a promising alternative approach for PcP diagnosis.
- [Show abstract] [Hide abstract] ABSTRACT: To analyze the direct medical costs of HIV/AIDS in Portugal from the perspective of the National Health Service. A retrospective analysis of medical records was conducted for 150 patients from five specialized centers in Portugal in 2008. Data on utilization of medical resources during 12 months and patients' characteristics were collected. A unit cost was applied to each care component using official sources and accounting data from National Health Service hospitals. The average cost of treatment was 14,277 €/patient/year. The main cost-driver was antiretroviral treatment (€ 9,598), followed by hospitalization costs (€ 1,323). Treatment costs increased with the severity of disease from € 11,901 (> 500 CD4 cells/µl) to € 23,351 (CD4 count ≤ 50 cells/ µl). Cost progression was mainly due to the increase in hospitalization costs, while antiretroviral treatment costs remained stable over disease stages. The high burden related to antiretroviral treatment is counterbalanced by relatively low hospitalization costs, which, however, increase with severity of disease. The relatively modest progression of total costs highlights that alternative public health strategies that do not affect transmission of disease may only have a limited impact on expenditure, since treatment costs are largely dominated by constant antiretroviral treatment costs.
- [Show abstract] [Hide abstract] ABSTRACT: Pneumocystis pneumonia (PcP) is a major cause of mortality and morbidity in immunocompromised patients. There are limited alternative therapeutic choices to trimethoprim-sulfamethoxazole (TMP-SMX) which is the standard first line therapy/prophylaxis for PcP. The efficacy of low doses of caspofungin and caspofungin in association with TMP-SMX standard-prophylactic dose was evaluated in an experimental model of Pneumocystis. Susceptibility of Pneumocystis spp. to low doses of caspofungin and caspofungin/TMP-SMX was evaluated in Balb/c immunosuppressed mice, infected intranasally with P. murina. Caspofungin was administered once daily at 0.1 mg/kg, 0.05 mg/kg, and 0.001 mg/kg and TMP-SMX was administered by oral gavage (12.25 mg/62.5 mg/day), for 21 days. Efficacy was calculated based on the reduction in organism burden determined through quantitative fluorescent-based real-time PCR (qPCR). Serum β-1,3-D-glucan was measured as an additional marker of infection. The present data showed that caspofungin demonstrated anti-Pneumomocystis effect. However, the doses administrated were too low to achieve Pneumocystis eradication, which suggests that echinocandin treatment should not be administrated as mono-therapy. After 21 days of treatment, P. murina was not detected in the lungs of mice with either TMP-SMX or caspofungin/TMP-SMX. The results showed that, even at the lowest concentrations tested, the efficacy of caspofungin in association with TMP-SMX was higher than the efficacy of either drug used alone. The administration of caspofungin/TMP-SMX was at least 1.4 times more effective against P. murina infection than TMP-SMX used alone. The most promising result was achieved with the combination of caspofungin 0.05 mg/kg/day with TMP-SMX 12.5 mg-62.5 mg/day, which reduced the parasite burden to undetectable levels immediately at the 14(th) day of treatment, showing a highly marked anti-Pneumomocystis effect. These data suggest that the administration of low doses of caspofungin in combination with low doses of TMP-SMX may provide an improved treatment protocol for Pneumocystis infection clearance.
- [Show abstract] [Hide abstract] ABSTRACT: BACKGROUND: Cobicistat (COBI) is a pharmacoenhancer with no antiretroviral activity in vitro. METHODS: An international, randomized, double-blind, double-dummy, active-controlled trial was conducted to evaluate the efficacy and safety of COBI versus ritonavir (RTV) as a pharmacoenhancer of atazanavir (ATV) in combination with emtricitabine (FTC)/tenofovir disoproxil fumarate (TDF) in treatment-naive patients. The primary end point was a human immunodeficiency virus type 1 (HIV-1) RNA load of <50 copies/mL at week 48 by the Food and Drug Administration snapshot algorithm; the noninferiority margin was 12%. RESULTS: A total of 692 patients were randomly assigned to a treatment arm and received study drug (344 in the COBI group vs 348 in the RTV group). At week 48, virologic success was achieved in 85% of COBI recipients and 87% of RTV recipients (difference, -2.2% [95% confidence interval, -7.4% to 3.0%]); among patients with a baseline HIV-1 RNA load of >100 000 copies/mL, rates were similar (86% vs 86%). Similar percentages of patients in both groups had serious adverse events (10% of COBI recipients vs 7% of RTV recipients) and adverse events leading to discontinuation of treatment with the study drug (7% vs 7%). Median increases in the serum creatinine level were 0.13 and 0.09 mg/dL, respectively, for COBI and RTV recipients. CONCLUSIONS: COBI was noninferior to RTV in combination with ATV plus FTC/TDF at week 48. Both regimens achieved high rates of virologic success. Safety and tolerability profiles of the 2 regimens were comparable. Once-daily COBI is a safe and effective pharmacoenhancer of the protease inhibitor ATV.
- [Show abstract] [Hide abstract] ABSTRACT: Introduction: Three major public health problems, tuberculosis, malaria and HIV/AIDS, are widespread in Angola, often as co-infections in the same individual. In 2009, it was assumed that 44,151 new cases of TB occurred in Angola. Interestingly, interventions such as treatment/prevention of malaria appear to reduce mortality in HIV-infected and possibly TB co-infected patients. However, despite the seriousness of the situation, current data on TB and co-infection rates are scarce. This study aimed to characterize all TB cases seen at the Hospital Sanatório de Luanda, and to determine the co-infection rate with HIV and/or malaria. Methodology: This retrospective study collected demographic, diagnostic and clinical data from all patients admitted during 2007. Results: A total of 4,666 patients were admitted, of whom 1,906 (40.8%) were diagnosed with TB. Overall, 1,111 patients (58.3%) were male and most patients (n=1302, 68.3%) were adults (≥ 14 years). The rate of HIV co-infection was 37.4% (n=712). Malaria was diagnosed during admission and hospital stay in 714 patients (37.5%), with Plasmodium falciparum the predominant species. Overall mortality was 15.2% (n=290). Conclusions: Because Luanda does not have the infrastructure to perform culture-based diagnosis of TB, confirmation of TB is problematic. The HIV-co-infection rate is high, with 37.4% of patients requiring integrated approaches to address this problem. With more than 1/3 of the TB patients co-infected with malaria, even during the hospital stay, the prevention of malaria in TB patients appears to be an effective way to reduce overall mortality.
- [Show abstract] [Hide abstract] ABSTRACT: Background Acute kidney injury (AKI) is common in hospitalized human immunodeficiency virus (HIV)-infected patients and is associated with hospital mortality. We aimed to evaluate the impact of AKI on long-term mortality of hospitalized HIV-infected patients. Methods Retrospective analysis of a cohort of 433 hospitalized HIV-infected patients who were discharged alive from the hospital. AKI was defined according to ‘Risk Injury Failure Loss of kidney function End-stage kidney disease’ creatinine criteria, as an increase of baseline serum creatinine (SCr) X 1.5 or in patients with baseline SCr > 4 mg/dL if there was an acute rise in SCr of at least 0.5 mg/dL. Cumulative mortality curves were determined by the Kaplan-Meier method, and log-rank test was employed to analyze statistically significant differences between curves. Cox regression method was used to determine independent predictors of mortality. Risk factors were assessed with univariate analysis, and variables that were statistically significant (P < 0.05) in the univariate analysis were included in the multivariate analysis. Results Sixty-four patients (14.8%) had AKI. Median follow-up was 37 months. At follow-up 81 patients (18.7%) died. At 1, 2 and 5 years of follow-up, the cumulative probability of death of patients with AKI was 21.2, 25 and 31.3%, respectively, as compared with 10, 13.3 and 16.5% in patients without AKI (log-rank, P = 0.011). In multivariate analysis AKI was associated with increased mortality (adjusted HR 1.7, 95% CI 1.1-3; P = 0.049). Conclusions AKI was independently associated with long-term mortality of hospitalized HIV-infected patients.
- [Show abstract] [Hide abstract] ABSTRACT: Toxoplasmosis is an important zoonosis worldwide. Here we determined the presence of Toxoplasma gondii antibodies in sera and T. gondii DNA in feces of 215 domestic cats from veterinary clinics in the Lisbon area; 44 (20.5%) had anti-T. gondii IgG antibodies by the modified agglutination test (cut-off 1:40) and DNA was detected in 16 (35.6%) of 45 cat feces tested. Risk factor analysis indicated increase of seroprevalence with age of the cats. Sera and tissues of 381 pigs from a slaughter house were also tested for T. gondii infection; 27 (7.1%) of the 381 pigs were seropositive. T. gondii DNA was demonstrated in diaphragms and/or brains of seven (35.0%) of 20 anti-T. gondii seropositive pigs tested by the B1 nested-PCR. Results indicate very high prevalence of T. gondii DNA in the feces (oocysts) of definitive hosts and relatively low, but still worrying, seroprevalence of T. gondii antibodies in pigs destined for human consumption.
- [Show abstract] [Hide abstract] ABSTRACT: Objectives: To characterize the nature and dynamics of the neutralizing antibody (NAb) response and escape in chronically HIV-2 infected patients. Methods: Twenty-eight chronically infected adults were studied over a period of 1-4 years. The neutralizing activity of plasma immunoglobulin G (IgG) antibodies against autologous and heterologous primary isolates was analyzed using a standard assay in TZM-bl cells. Coreceptor usage was determined in ghost cells. The sequence and predicted three-dimensional structure of the C2V3C3 Env region were determined for all isolates. Results: Only 50% of the patients consistently produced IgG NAbs to autologous and contemporaneous virus isolates. In contrast, 96% of the patients produced IgG antibodies that neutralized at least two isolates of a panel of six heterologous R5 isolates. Breadth and potency of the neutralizing antibodies were positively associated with the number of CD4(+) T cells and with the titer and avidity of C2V3C3-specific binding IgG antibodies. X4 isolates were obtained only from late stage disease patients and were fully resistant to neutralization. The V3 loop of X4 viruses was longer, had a higher net charge, and differed markedly in secondary structure compared to R5 viruses. Conclusion: Most HIV-2 patients infected with R5 isolates produce C2V3C3-specific neutralizing antibodies whose potency and breadth decreases as the disease progresses. Resistance to antibody neutralization occurs in late stage disease and is usually associated with X4 viral tropism and major changes in V3 sequence and conformation. Our studies support a model of HIV-2 pathogenesis in which the neutralizing antibodies play a central role and have clear implications for the vaccine field.
- [Show abstract] [Hide abstract] ABSTRACT: Purpose of the study: About a third of HIV-infected patients in Europe present for care late in their disease. In 2010 a consensus was reached for a definition of late presentation, providing a new tool for research and acting. With regard to HIV2 infection this population has never been characterized. Methods: Retrospective analysis of the clinical records of all the HIV2-infected patients who meet the inclusion criteria (those defined by the European Late Presenter Consensus working group: Late presentation of HIV infection: a consensus definition 1) admitted to our ward between January 2006 and December 2011. The patients were characterized according to epidemiological, clinical and immunologic status and outcome. Summary of results: During the period analyzed, 15 HIV2 patients were late presenters. The mean age of the patients was 48 years old (although 53% were older than 50 years); 8 (53%) were men; 11 (73%) were of African origin. Heterosexual transmission was reported in three of the patients, in the remainder the transmission mode was not available. The mean TCD4 cell was 188 (range 27-339), with 8 (53%) with a CD4 count below 200 cells. Twelve (80%) of the patients fulfilled AIDS criteria. There were 3 deaths, corresponding to a mortality rate of 20%. The cause of death was disseminated tuberculosis in two cases and non-Hodgkin's lymphoma in the third case. Conclusions: Most late presenters with HIV2 infection are of African origin, there is an even distribution between genders, their mean age is around 50 years old, more than half had a CD4 cell count below 200 and there was a 20% mortality rate. These patients pose challenges at various levels: their mortality rate is much higher than in the general HIV-infected population and they are diagnosed very late, leading to a disproportionate increase in risk of transmission, morbidity and mortality.
- [Show abstract] [Hide abstract] ABSTRACT: Purpose of the study: This study intends to characterize a Portuguese patient population with chronic HCV and HIV coinfection, followed at our Research Unit, underline the importance of early treatment and incorporate the importance of DDA for retreatment of HCV infection. Methods: Retrospective, observational analysis of medical records of 348 HCV/HIV coinfected patients from 2001 to 2011. Demographic, epidemiological, clinical and laboratory data and virologic response were collected. Summary of results: Review of 348 HCV/HIV coinfected patients, 121 of those (34.7%) under treatment, predominantly male (77.0%) and Caucasians (94.8%) with a median age of 44 yrs old (min 25; max 77 yrs). Intravenous drug use was the main route of HCV infection, in 71.3% of patients, and 8.3% were related with MSM. Frequent morbidities were alcohol abuse (46.8%), illicit drug use (70.1%), methadone (25.6%) and mental disturbances (12.3%) of patients. Regarding HIV infection, six were HIV-2 and 342 HIV-1; 36.1% were stage A and 29.6% were stage C (CDC Atlanta), 94.8% on antiretroviral treatment and only 21.9% of them with more than 350 TCD4 cell count. Genotype 1 was the most prevalent (58.1%-117 genotype 1a, 26 genotype 1b); 1.6% were genotype 2, 22.8% genotype 3 and 17.5% genotype 4. Previous to treatment initiation, HCV ARN was above 600.000 IU/mL in 56.9% patients. Fibrosis was evaluated by fibroelastography in 41.1% and hepatic biopsy in 26.3% of patients; in those, 44.0% had a score above F2 (METAVIR) and ALT was elevated 2 times the limit in 38.0%, with an average value of 94 UI/L. IL 28B testing was performed in only 35 patients at the time, with 45.7% CC and 17.1% CT genotype. Treatment was started in 34.8% of patients, with 1.7 treatments per individual, and regimen was based on peguilated interferon with ribavirin in 93.6% of cases (72.1% with peginterferon alfa 2a). The SVR rate was 51.2%, with 28.9% non responders, 3 relapsers and 9 treatment interruptions due to major toxicities. Conclusions: Our data presents a low HCV treatment initiation, illustrated by 65.2% patients who did not begin any treatment. The majority completed treatment and the SVR rate was similar to literature. Individualized approach is essential to determine the optimal time to initiate HCV treatment, to assess patient adherence and adverse events management, in order to optimize treatment and reserve DDA drugs to experienced patients with worse predictive factors.
- [Show abstract] [Hide abstract] ABSTRACT: Pneumocystis has been identified in various mammalian species, including domestic, wild and zoo animals. This study's main objectives were: (1) to estimate the prevalence of the Pneumocystis carinii f. sp. suis infection in slaughtered pigs in Portugal, (2) assess the prevalence differences within distinct age groups of animals, (3) determine the possible associations between pulmonary lesions and the infection, and (4) genetically characterize the P. carinii f. sp. suis isolates recovered from infected animals using PCR with DNA sequencing. An epidemiological cross-sectional study was conducted using 215 pig lung tissue samples which demonstrated a global prevalence of 7% (14 positive samples). This value was later validated by statistical analysis as being representative of the national population prevalence. Regarding the assessment of relations between the different variables investigated during the study (age, gender, geographical region, type of farming, weight and pulmonary lesion) and the P. carinii f. sp. suis infection, no significant statistical differences were found, and apparently, no predisposing factors could be defined. Nevertheless, infection by Pneumocystis in pigs is ubiquitous and it can be detected in healthy animals. Thus, the colonization of P. carinii f. sp. suis among healthy individuals suggests that asymptomatic carriers can be an effective reservoir for susceptible animals and participate in the transmission of infection. The present data confirmed that porcine Pneumocystis is genetically distinct from Pneumocystis DNA detected in other mammalian hosts.
- [Show abstract] [Hide abstract] ABSTRACT: Pneumocystis jirovecii (Pj) is an opportunistic pathogen responsible for one of the most severe infections in immunocompromised patients, Pneumocystis pneumonia (PcP). Currently, the laboratory diagnosis of PcP is based on cytochemical staining, indirect immunofluorescence with monoclonal antibodies and PCR on induced sputum and bronchoalveolar lavage samples. Studies showed that Pneumocystis species have a need for exogenous S-adenosyl Methionine (SAM) since they are unable to synthetize this compound. Other studies suggest that Krebs von den Lungen-6 (KL-6), a mucin-like glycoprotein expressed on type II alveolar pneumocytes and bronchiolar epithelial cells, may be used as a sensitive indicator of various types of interstitial pneumonitis. The aim of the present work is to evaluate these two potential serological markers (SAM and KL- 6) for PcP diagnosis. Sera and pulmonary specimens were obtained from 65 HIV+ and seven HIV- Portuguese patients for diagnostic purpose (1997-2012). Sera were collected from 25 blood donors (negative control group). PcP cases were confirmed by using indirect immunofluorescence with monoclonal antibodies (IF) and nested-PCR. Of the 72 patients studied, 47 were PcP positive (median: SAM 38 nmol/L; KL-6 1416 U/mL) and 25 were PcP negative (10 were colonized by Pj presenting subclinical infection and 15 were not infected by Pj) (median: SAM 36 nmol/L; KL-6 556 U/mL). The median serologic markers levels detected for the colonized carriers were 21 nmol/L (SAM) and 542 U/mL (KL-6), while in the patients without Pj infection were 60nmol/L (SAM) and 670 U/mL (KL-6). In the blood donors the median SAM serological level was 75 nmol/L, while the median KL-6 level was 459 U/mL. Sera presenting high levels of KL-6 (>570 U/mL) were statistically associated with diagnosed PcP cases (77%, P<0.001). These preliminary results suggest that KL-6 might be a candidate to use in PcP diagnosis, mostly when used in association with other serologic markers. Supported by “Centro de Malária e Outras Doenças Tropicais (CMDT)”.
Hospital de Santa MariaLisboa, Lisbon, Portugal
Copenhagen University Hospital HvidovreHvidovre, Capital Region, Denmark
Instituto de Salud Carlos IIIMadrid, Madrid, Spain
University of LisbonLisboa, Lisbon, Portugal