Resumo em Inglês:

Abstract Background Development of drug-resistance mutations is the main cause of failure in antiretroviral therapy. In Brazil, there is scarce information on resistance pattern for patients failing antiretroviral therapy. Objectives To define the HIV mutational profile associated with drug resistance in Brazilian patients from 5 large cities, after first, second or further failures to antiretroviral therapy. Methods We reviewed genotyping results of 1520 patients failing therapy in five Brazilian cities. Frequency of mutations, mean number of active drugs, viral susceptibility to each antiretrovirals drug, and regional differences were assessed. Results Mean time of antiretrovirals use was 22.7 ± 41.1 months. Mean pre-genotyping viral load was 4.2 ± 0.8 log (2.1 ± 2.0 after switching antiretrovirals). Mean number of remaining active drugs was 9.4, 9.0, and 7.9 after 1st, 2nd, and 3rd failure, respectively. We detected regional variations in drug susceptibility: while BA and RS showed the highest (∼40%) resistance level to ATV/r, FPV/r and LPV/r, in the remaining cities it was around half of this rate. We detected 90% efavirenz/nevirapine resistance in SP, only 45% in RS, and levels between 25% and 30% in the other cities. Regarding NRTI, we found a similar pattern, with RJ presenting the highest, and CE the lowest susceptibility rates for all NRTI. Zidovudine resistance was detected in only 3% of patients in RJ, against 45–65% in the other cities. RJ and RS showed 3% resistance to tenofovir, while in CE it reached 55%. DRV/r (89–97%) and etravirine (61–85%) were the most active drugs, but again, with a wide variation across cities. Conclusions The resistance mutational profile of Brazilian patients failing antiretroviral therapy is quite variable, depending on the city where patients were tested. This variation likely reflects distinctive choice of antiretrovirals drugs to initiate therapy, adherence to specific drugs, or circulating HIV-1 strains. Overall, etravirine and DRV/r remain as the most active drugs.

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Abstract Introduction Worldwide about 30% of HIV-infected patients are coinfected with HCV or HBV. The HIV/HCV coinfection is more common in individuals who have a history of drug addiction. The aims of this study were to assess the HCV and HBV prevalence in HIV-infected patients and analyze their characteristics. Methods We considered the new HIV diagnoses notified by the regional surveillance system of Tuscany from 2009 to 2013. Descriptive analyses were conducted on the socio-demographic characteristics, routes of transmission, and reason to perform the test. In coinfected patients we assessed the risk for being late presenter (LP) or the risk of having AIDS. Results In 5 years of surveillance a total of 1354 new HIV diagnoses were notified: 1188 (87.7%) were HIV alone, 106 (7.8%) HIV/HCV, 56 (4.1%) HIV/HBV, and 4 (0.33%) HIV/HCV/HBV. The main risk factor was injection drug use in 52.8% of HCV/HIV cases, while in HIV/HBV patients the main risk factor was sexual exposure. HIV/HBV coinfected patients showed worse clinical and immunological features than HIV and HIV/HCV patients: 78.6% had CD4 count less than 350 mm−3 (vs. 54.6% and 62.1%, respectively) and 39.4% had AIDS (vs 20.7% and 7.6%). The risk for being LP triples for HIV/HBV (OR 2.98; 95% IC: 1.56–5.70) than patients with HIV alone. Conclusions We have observed less advanced disease in HIV and HCV-HIV patients compared with HBV–HIV coinfected patients. Moreover, our results show a higher prevalence of HIV/HCV among drug addicts and in the age-group 35–59, corresponding to those born in years considered most at risk for addiction. This study also confirms the finding of a less advanced HIV disease in HIV/HCV coinfected patients.

Resumo em Inglês:

Abstract Background Several in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans. Methods A total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae. Results All used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains. Conclusions The in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.

Resumo em Inglês:

Abstract A natural and biocompatible fibrin microsphere is one of the most promising dual delivery vehicle as compared to other traditionally designed delivery modalities. It represents sustained delivery of encapsulated drug and is easily biodegradable in the blood circulation. In the present study, we evaluated the systemic augmentation of the antifungal activity of amphotericin B loaded in fibrin microsphere (AMB-fibrin microsphere) against cryptococcosis in Swiss albino mice. Mice infected with Cryptococcus neoformans were treated with 0.5 mg/kg AMB-fibrin microsphere that was given alternately for 7 days. The antifungal potential of AMB-fibrin microsphere was assessed on the basis of reduction of cfu count in the systemic circulation and various vital organs of infected mice. The formulation was found to be highly effective in reducing intracellular pathogen from the experimental animals where fibrin microsphere significantly controlled the release of amphotericin B for longer time duration. The AMB-fibrin microsphere chemotherapy was significantly more effective than free amphotericin B in reducing the fungal burden and showed better survival efficacy (p < 0.05). The current study demonstrating the use of novel amphotericin B loaded fibrin microsphere not only imparts protection to the encapsulated amphotericin B but also offers an effective strategy to decrease the drug associated toxicities.

Resumo em Inglês:

Abstract Background The Roma ethnic group is the largest and most marginalized minority in Europe, believed to be vulnerable to sexually transmitted infections. Aim The purpose of the study was to investigate frequency and characteristics of gonorrhea and syphilis among the Roma population in Belgrade. Methods Data from the City Institute for Skin and Venereal Diseases to which all gonorrhea and syphilis cases are referred were analyzed. Results During the period of 2010–2014 sexually transmitted infections were more frequent among Roma than in rest of Belgrade population. Average percentages of Roma among all reported subjects with syphilis and those with gonorrhea were 9.6% and 13.5%, respectively, while the percentage of Roma in the total Belgrade population was about 1.6%. Roma with syphilis and gonorrhea were more frequently men (75%), most frequently aged 20–29 years (43.4%), never married (64.5%), with elementary school or less (59.2%), unemployed (80.3%), and heterosexual (89.5%). Among Roma 10.5% were sex workers and 68.4% did not know the source of their infection. Significant differences between Roma cases and other cases in Belgrade in all characteristics observed were in agreement with differences between Roma population and the total population of Serbia. Conclusion The present study confirmed the vulnerability of the Roma population to sexually transmitted infections.

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Abstract Introduction Invasive mold disease is an important complication of patients with hematologic malignancies, and is associated with high mortality. A diagnostic-driven approach has been an alternative to the classical empiric antifungal therapy. In the present study we tested an algorithm that incorporated risk stratification using the D-index, serial serum galactomannan and computed tomographic-scan to guide the decision to start antifungal therapy in neutropenic patients. Patients and methods Between May 2010 and August 2012, patients with acute leukemia in induction remission were prospectively monitored from day 1 of chemotherapy until discharge or death with the D-index and galactomannan. Patients were stratified in low, intermediate and high risk according to the D-index and an extensive workup for invasive mold disease was performed in case of positive galactomannan (≥0.5), persistent fever, or the appearance of clinical manifestations suggestive of invasive mold disease. Results Among 29 patients, 6 (21%), 11 (38%), and 12 (41%) were classified as high, intermediate, and low risk, respectively. Workup for invasive mold disease was undertaken in 67%, 73% and 58% (p = 0.77) of patients in each risk category, respectively, and antifungal therapy was given to 67%, 54.5%, and 17% (p = 0.07). Proven or probable invasive mold disease was diagnosed in 67%, 45.5%, and in none (p = 0.007) of high, intermediate, and low risk patients, respectively. All patients survived. Conclusion A risk stratification using D-index was a useful instrument to be incorporated in invasive mold disease diagnostic approach, resulting in a more comprehensive antifungal treatment strategy, and to guide an earlier start of treatment in afebrile patients under very high risk.

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Abstract JC virus (JCV) is a member of the Polyomaviridae family and is associated to a severe disease known as progressive multifocal leukoencephalopathy, PML, which is gradually increasing in incidence as an opportunistic infection among AIDS patients. The present study aimed to investigate the occurrence of JCV among HIV-1 carriers including their types and molecular subtypes and the possible association with disease. Urine samples from 66 HIV-1 infected subjects were investigated for the presence of the virus by amplifying VP1 (215 bp) and IG (610 bp) regions using the polymerase chain reaction. JCV was detected in 32% of the samples. The results confirmed the occurrence of type B (subtype Af2); in addition, another polyomavirus, BKV, was also detected in 1.5% of samples of the HIV-1 infected subjects. Apparently, there was no significant difference between mono- (HIV-1 only) and co-infected (HIV-1/JCV) subjects regarding their TCD4+/TCD8+ lymphocyte counts or HIV-1 plasma viral load. Self admitted seizures, hearing and visual loses were not significantly different between the two groups.

Resumo em Inglês:

Abstract Aims We sought to characterize the antibiotic susceptibility of strains of Stenotrophomonas maltophilia isolated from clinical samples, and the role of Stenotrophomonas maltophilia biofilm in antibiotic resistance. Methods Fifty-one clinical Stenotrophomonas maltophilia isolates were obtained from patients with nosocomial infection in the surgical wards and ICUs of six general hospitals in Tianjin, China. In vitro models of Stenotrophomonas maltophilia biofilms were established and confirmed by scanning electron microscopy and fluorescence microscopy with silver staining. The minimal inhibitory concentrations and biofilm inhibitory concentrations of commonly used antibiotics were determined. Results 47 of 51 strains were resistant to three or more antibiotics. 42 of 51 strains formed Stenotrophomonas maltophilia biofilms in vitro. Stenotrophomonas maltophilia biofilm formation greatly reduced sensitivity to most tested antibiotics, but not to levofloxacin. However, in the presence of erythromycin scanning electron microscopy revealed that levofloxacin inhibited Stenotrophomonas maltophilia biofilm formation. Factorial ANOVA revealed that erythromycin enhanced susceptibility to levofloxacin, cefoperazone/sulbactam, and piperacillin (p < 0.05), and an ΔE model revealed that levofloxacin and erythromycin acted synergistically in biofilms, suggesting specific use of combined macrolide therapy may represent an effective treatment for Stenotrophomonas maltophilia infection. Conclusions Antibiotics could act synergistically to combat the protection conferred to clinical isolates of Stenotrophomonas maltophilia by biofilms. Macrolide antibiotics may be effective where used in combination.

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Abstract Objectives This retrospective study was aimed to explore the epidemiological and clinical profiles of Mycoplasma pneumoniae infection in neonates. Methods From 2011 to 2014, 1322 hospitalized neonates with lower respiratory tract infections were screened for Mycoplasma pneumoniae by detection of Mycoplasma pneumoniae antibodies using Serion ELISA classic Mycoplasma pneumoniae kits. Results Mycoplasma pneumoniae was identified in 89 (6.7%) patients. The age ranged from 1 day to 28 days with a median of 22 days. The male to female ratio was 1.15:1. Mycoplasma pneumoniae infection peaked in spring (from March through May) and winter (from December through February). Compared with non-Mycoplasma pneumoniae infected neonates, those with Mycoplasma pneumoniae infection were older, presented fever more frequently, and had less tachypnea. Conclusions Mycoplasma pneumoniae could be an important etiologic agent for respiratory tract infection in neonates. In neonates Mycoplasma pneumoniae infection was usually associated with older age, presence of fever, and less tachypnea. Mycoplasma pneumoniae infection in neonates tends to be a mild process.

Resumo em Inglês:

Abstract Background In the current study we aimed to find out the impact of cytokine-inducible Src homology 2 domain protein (CISH) gene polymorphisms on the risk of pulmonary tuberculosis (PTB) in a sample of Iranian population. Materials and methods Polymorphisms of CISH rs2239751, rs414171, and rs6768300 were determined in 200 PTB patients and 200 healthy subjects using T-ARMS-PCR or PCR-RFLP method. Results The results showed that rs414171 A>T genotypes significantly decreased the risk of PTB (OR = 0.16, 95% CI = 0.10–0.27, p < 0.0001, AT vs AA; OR = 0.31, 95% CI = 0.14–0.68, p < 0.0001, TT vs AA; OR = 0.19, 95% CI = 0.12–0.29, p < 0.0001, AT+TT vs AA; OR = 0.29, 95%CI = 0.20–0.42, p < 0.0001, T vs A). For rs6768300, the findings indicated that this variant decreased the risk of PTB (OR = 0.52, 95% CI = 0.33–0.82, p = 0.005, CG vs GG; OR = 0.57, 95% CI = 0.38–0.87, p = 0.012, C vs G). No significant association was observed between CISH rs2239751 polymorphism and risk/protection of PTB. Conclusion Our findings indicated that CISH rs414171 and rs6768300 variants might be associated with protection from PTB.

Resumo em Inglês:

Abstract Infection caused by carbapenem-resistant Klebsiella pneumoniae has become a major healthcare threat and KPC-2 enzyme is a dominant factor mediating carbapenems resistance in K. pneumoniae. This study was designed to determine the genetic environment of bla KPC-2, which prevailed in clinical K. pneumoniae isolates recovered in Huashan Hospital, Shanghai, China. Forty-two clinical isolates were included in this study by bla KPC-2 screening. After multilocus sequence typing and plasmid analyses of PCR-based replicon typing (PBRT), junction PCR, mapping PCR and crossing PCR assays, primer walking, and amplicon sequencing were used to analyze the genetic environment of the bla KPC-2 gene. ST423, ST65, ST977, and ST11 were all detected in KPC-2-producing K. pneumoniae. Two types of bla KPC-2-bearing genetic structure were found: Tn1721-bla KPC-2-Tn3 and Tn1721-bla KPC-2-ΔTn3-IS26; and were carried in IncX and IncFII plasmids, respectively. In conclusion, the genetic environment of the bla KPC-2 gene was diverse and Tn1721-bla KPC-2-ΔTn3-IS26 was dominant in clinical K. pneumoniae isolates in Huashan Hospital. This study sheds some light on the genetic environment and should foster further studies about the mechanism of the bla KPC-2 dissemination.

Resumo em Inglês:

Abstract Introduction Fusarium spp are ubiquitous fungi recognized as opportunistic agents of human infections, and can produce severe infections in burn patients. The literature on Fusarium spp infections in pediatric burn patients is scarce. Objectives To describe the clinical and epidemiological features as well as outcome of Fusarium spp infections in pediatric burn patients. Patients and methods Retrospective, descriptive study of Fusarium spp infections in a specialized intensive care burn unit. Results In 15 patients Fusarium spp infections were diagnosed. Median age was 48 months. Direct fire injury was observed in ten patients. The median affected burn surface area was 45%. Twelve patients had a full thickness burn. Fourteen patients had a Garces Index ≥3. Fungal infection developed at a median of 11 days after burn injury. Fungi were isolated from burn wound in 14 patients and from the bone in one patient. Amphotericin B was the drug of choice for treatment followed by voriconazole. Median time of treatment completion was 23 days. One patient (7%) died of fungal infection-related causes. Conclusion In our series Fusarium spp was an uncommon pathogen in severely burnt patients. The burn wound was the most common site of infection and mortality was low.

Resumo em Inglês:

Abstract Syphilis is an infectious disease caused by the bacterium Treponema pallidum. Syphilis has three clinical stages and may present various oral manifestations, mainly at the secondary stage. The disease mimics other more common oral mucosa lesions, going undiagnosed and with no proper treatment. Despite the advancements in medicine toward prevention, diagnosis, and treatment syphilis remains a public health problem worldwide. In this sense, dental surgeons should be able to identify the most common manifestations of the disease in the oral cavity, pointing to the role of this professional in prevention and diagnosis. This study describes a case series of seven patients with secondary syphilis presenting different oral manifestations.