RESUMO
The aims of this study were to evaluate the epidemiology of nosocomial candidemia in a large teaching hospital in Brescia, Italy, and the in vitro antifungal susceptibility of isolates. We analyzed 196 isolates causing fungemia in patients admitted in our hospital, between January 2009 and December 2015. Strains were identified by VITEK 2 and MALDI-TOF MS. MICs were determined by Sensititre Yeast OneTM. The resistance was defined by using the revised CLSI breakpoints/epidemiological cutoff values to assign susceptibility or wild type to systemic antifungal agents. Most infections were caused by Candida albicans (60%), Candida parapsilosis (15%), Candida glabrata (12%) and Candida tropicalis (6%). The susceptibility rate for fluconazole was 96.5%. Non-Candida species isolates exhibited full susceptibilities to echinocandins according to CLSI breakpoints. Amphotericin B demonstrated excellent activity against all Candida species. Local epidemiological and antifungal susceptibility studies are necessary in order to improve empirical treatment guidelines.
Assuntos
Antifúngicos/farmacologia , Candida/classificação , Candida/efeitos dos fármacos , Candidíase Invasiva/epidemiologia , Candidíase Invasiva/microbiologia , Farmacorresistência Fúngica , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anfotericina B/farmacologia , Candida/isolamento & purificação , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Equinocandinas/farmacologia , Feminino , Fluconazol/farmacologia , Hospitais de Ensino , Humanos , Itália/epidemiologia , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Técnicas de Tipagem Micológica , Estudos Retrospectivos , Adulto JovemRESUMO
Of 901 group B streptococcus strains analyzed, 13 (1.4%) were resistant to levofloxacin (MICs of >32 µg/ml for seven isolates, 2 µg/ml for four isolates, and 1.5 µg/ml for four isolates). Mutations in the quinolone resistance-determining regions (QRDRs) of gyrase and topoisomerase IV were identified. A double mutation involving the Ser-81 change to Leu for gyrA and the Ser-79 change to Phe or to Tyr for parC was linked to a high level of fluoroquinolone resistance. In addition, two other mutational positions in parC were observed, resulting in an Asp-83-to-Tyr substitution and an Asp-83-to-Asn substitution. Different mutations were also observed in gyrB, with unknown significance. Most levofloxacin-resistant GBS strains were of serotype Ib and belonged to sequence type 19 (ST19) and clonal complex 19 (CC-19). Most of them exhibited the epsilon gene.
Assuntos
Antibacterianos/farmacologia , Levofloxacino/farmacologia , Streptococcus agalactiae/efeitos dos fármacos , DNA Girase/genética , DNA Topoisomerase IV/genética , DNA Topoisomerases Tipo I/genética , Farmacorresistência Bacteriana/genética , Itália , Testes de Sensibilidade Microbiana , Mutação , Streptococcus agalactiae/genéticaRESUMO
BACKGROUND AND AIM: There is uncertainty regarding the prevention of migraine crises by changing the lifestyle of patients. The aim of this randomized, crossover intervention trial was to evaluate the effects of a low lipid intake on the incidence and severity of migraine crises, in comparison to a diet with moderate lipid intake. METHODS AND RESULTS: After a 2-month run-in when patients received preventive medication but were left on their habitual diet, a low-lipid or a normal-lipid diet was randomly prescribed for 3 months and thereafter diets were crossed over for the following 3 months. Headache was diagnosed based on the International Classification of Headache Disorders (IHCD) III criteria. The number and severity of attacks were assessed using a self-reported calendar. Adherence to the diet was assessed by a food frequency questionnaire. An analysis was performed on the 83 episodic or chronic migraineurs (63 female and 20 male), in the age range of 18-57 years, who completed both intervention periods. Obese subjects had a significantly higher number of attacks than those overweight or with normal body weight (24.7 ± 8, 16.3 ± 12, and 15.6 ± 11, respectively, p < 0.03) with a significant relationship between the body mass index (BMI) and the number of monthly attacks (r = 0.238, p < 0.03). The number (2.9 ± 3.7 vs. 6.8 ± 7.5, p < 0.001) and severity (1.2 + 0.9 vs. 1.7 ± 0.9, p < 0.01) of attacks significantly decreased during both intervention periods, with a significant difference in favour of the low-lipid diet. CONCLUSIONS: In this group of patients, the low-lipid diet significantly affected the number and severity of migraine attacks in comparison to a normal-lipid diet. ClinicalTrials.gov Identifier: NCT 01917474.
Assuntos
Gorduras na Dieta/administração & dosagem , Transtornos de Enxaqueca/dietoterapia , Adolescente , Adulto , Pressão Sanguínea , Índice de Massa Corporal , Peso Corporal , Estudos Cross-Over , Dieta com Restrição de Gorduras , Ingestão de Energia , Ácidos Graxos/administração & dosagem , Ácidos Graxos Monoinsaturados/administração & dosagem , Comportamento Alimentar , Feminino , Humanos , Estilo de Vida , Masculino , Pessoa de Meia-Idade , Estado Nutricional , Obesidade/dietoterapia , Azeite de Oliva/administração & dosagem , Adulto JovemRESUMO
PURPOSE: We have developed a sequencing assay for determining the usage of the genotypic HIV-1 co-receptor using peripheral blood mononuclear cell (PBMC) DNA in virologically suppressed HIV-1 infected patients. Our specific aims were to (1) evaluate the efficiency of V3 sequences in B versus non-B subtypes, (2) compare the efficiency of V3 sequences and tropism prediction using whole blood and PBMCs for DNA extraction, (3) compare the efficiency of V3 sequences and tropism prediction using a single versus a triplicate round of amplification. RESULTS: The overall rate of successful V3 sequences ranged from 100 % in samples with >3,000 copies HIV-1 DNA/10(6) PBMCs to 60 % in samples with <100 copies total HIV-1 DNA /10(6) PBMCs. Analysis of 143 paired PBMCs and whole-blood samples showed successful V3 sequences rates of 77.6 % for PBMCs and 83.9 % for whole blood. These rates are in agreement with the tropism prediction obtained using the geno2pheno co-receptor algorithm, namely, 92.1 % with a false-positive rate (FPR) of 10 or 20 % and of 96.5 % with an FPR of 5.75 %. The agreement between tropism prediction values using single versus triplicate amplification was 98.2 % (56/57) of patients using an FPR of 20 % and 92.9 % (53/57) using an FPR of 10 or 5.75 %. For 63.0 % (36/57) of patients, the FPR obtained via the single amplification procedure was superimposable to all three FPRs obtained by triplicate amplification. CONCLUSIONS: Our results show the feasibility and consistency of genotypic testing on HIV-1 DNA tropism, supporting its possible use for selecting patients with suppressed plasma HIV-1 RNA as candidates for CCR5-antagonist treatment. The high agreement between tropism prediction by single and triple amplification does not support the use of triplicate amplification in clinical practice.
Assuntos
Técnicas de Genotipagem/métodos , Infecções por HIV/virologia , HIV-1/genética , HIV-1/fisiologia , Técnicas de Diagnóstico Molecular/métodos , Receptores de HIV/metabolismo , Tropismo Viral , Adulto , DNA Viral/química , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Infecções por HIV/diagnóstico , HIV-1/classificação , HIV-1/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Provírus/classificação , Provírus/genética , Provírus/isolamento & purificação , Análise de Sequência de DNA , Internalização do VírusRESUMO
The purpose of this investigation was to analyse Streptococcus agalactiae (group B Streptococcus, GBS) isolates collected in Italy from vaginal and urine samples in respect to their clonality, distribution of virulence factors and antimicrobial resistance determinants. Three hundred and eighty-eight GBS were recovered from clinical samples. They were analysed for antibiotic resistance profiling. Erythromycin-resistant strains were further characterised by multilocus sequence typing (MLST), serotyping and the detection of alp genes of the alpha-like protein (Alp) family. GBS isolates represented 40 different sequence types (STs), grouped in five clonal complexes (CCs) and belonged to seven serotypes. Most serotype V strains (81%) possessed alp2-3; serotype Ia carried mainly epsilon, while the serotype III mainly rib. All isolates were susceptible to penicillin, whereas resistance to erythromycin was detected in 15% of isolates. Most erythromycin-resistant GBS strains were of serotype V (56.8%) and belonged to the CC-1 group (50%). Macrolide resistance phenotypes were the cMLS(B) (46.5%) and the M phenotypes (46.5%) due to the presence of ermB and mefA/E genes, respectively. These results provide data which establish a baseline for monitoring erythromycin resistance in this region and also provide an insight into the correlation among clonal types, serotypes, surface protein and resistance genes. The increased prevalence of strains that displayed the M phenotype strengthens the importance of the epidemiological surveillance of macrolide resistance in GBS, which may also represent an important reservoir of resistance genes for other species.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Eritromicina/farmacologia , Infecções Estreptocócicas/epidemiologia , Infecções Estreptocócicas/microbiologia , Streptococcus agalactiae/classificação , Streptococcus agalactiae/efeitos dos fármacos , Adulto , Idoso , Análise por Conglomerados , Feminino , Genes Bacterianos , Genótipo , Humanos , Itália/epidemiologia , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Epidemiologia Molecular , Tipagem Molecular , Fenótipo , Gravidez , Prevalência , Sorotipagem , Streptococcus agalactiae/genética , Streptococcus agalactiae/fisiologia , Urina/microbiologia , Vagina/microbiologia , Fatores de Virulência/genéticaRESUMO
Mycosis fungoides (MF) is a rare form of cutaneous T cell lymphoma suspected of having a viral etiology. As in adult T cell leukemia, the virus involved may be human T lymphotropic virus type 1 (HTLV-1). We cultured the peripheral blood mononuclear cells (PBMC) of 29 patients with MF HTLV-1 seronegative by enzyme-linked immunosorbent assay and Western blot. The presence of reverse transcriptase (RT) and p24 antigen was investigated in the concentrate supernatant of the culture. The DNA of all studied patients was submitted to polymerase chain reaction and Southern blot analysis using primers and probes recognizing the tax region of HTLV-1/2 and the pol region of HTLV-1. 10 of 29 patients were found positive to HTLV-1, whereas they were always negative to RT and p24. The same results were confirmed in double blind after 6 mo. Our findings suggest HTLV-1 may be involved in the etiology of MF, at least in certain cases.
Assuntos
DNA Viral/análise , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Leucócitos Mononucleares/virologia , Micose Fungoide/virologia , Neoplasias Cutâneas/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , Células Cultivadas , Genes pX , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/isolamento & purificação , Vírus Linfotrópico T Tipo 1 Humano/genética , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Micose Fungoide/sangue , Reação em Cadeia da Polimerase , Neoplasias Cutâneas/sangueAssuntos
Listeria monocytogenes/isolamento & purificação , Listeriose/epidemiologia , Listeriose/patologia , Aborto Espontâneo/diagnóstico , Aborto Espontâneo/microbiologia , Aborto Espontâneo/patologia , Adulto , Idoso , Bacteriemia/diagnóstico , Bacteriemia/microbiologia , Bacteriemia/patologia , Análise por Conglomerados , Infecção Hospitalar/diagnóstico , Infecção Hospitalar/microbiologia , Infecção Hospitalar/patologia , Feminino , Genótipo , Humanos , Itália/epidemiologia , Listeria monocytogenes/classificação , Listeria monocytogenes/genética , Listeriose/microbiologia , Masculino , Pessoa de Meia-Idade , Tipagem Molecular , Peritonite/diagnóstico , Peritonite/microbiologia , Peritonite/patologia , Reação em Cadeia da Polimerase , GravidezRESUMO
This study assessed changes in prevalence and distribution of HIV-1 non-subtype B viruses in Italian and immigrant patients over two decades in a province in Italy. All HIV-positive patients who underwent genotypic resistance testing were selected. Prevalence of non-subtype B viruses in 3-year periods was calculated. All sequences of non-subtype B and those provided by REGA as unassigned were analysed for phylogenetic relationships. In total, 250/1563 (16%) individuals were infected with a non-subtype B virus. Prevalence increased over time, reaching a peak (31.5%) in 2004-2006. In Italian patients, the most frequent subtypes were B (92.5%) and F1 (4%). F1 subtype was also prevalent in patients from South America (13.6%); in patients of African origin, CRF02_AG (54.9%) and G (12.3%) were the most frequent. HIV-1 non-subtype B infections in Italians were mostly found in patients who acquired HIV sexually. A phylogenetic relationship between F subtypes in Italian and representative HIV-1 sequences from Brazil was found. C subtypes in Italians were phylogenetically related to subtypes circulating in Brazil. Inter-subtype recombinants were also found in the latest years. The HIV-1 epidemic in Brescia province evolved to the point where about 1/3 patients recently diagnosed harboured non-B HIV subtypes. The distribution of HIV-1 non-B subtypes in Italian patients resembled that in South American patients and phylogenetic relatedness between some Italian and South American HIV-1 strains was found. The possible epidemiological link between these two populations would have been missed by looking only at risk factors for HIV acquisition declared by patients. The evidence of inter-subtype recombinants points to significant genetic assortment. Overall our results support phylogenetic analysis as a tool for epidemiological investigation in order to guide targeted prevention strategies.
Assuntos
Infecções por HIV/epidemiologia , Infecções por HIV/virologia , HIV-1/classificação , Adulto , Distribuição de Qui-Quadrado , Feminino , Genótipo , Infecções por HIV/etnologia , Infecções por HIV/genética , HIV-1/genética , Humanos , Itália/epidemiologia , Modelos Logísticos , Masculino , Epidemiologia Molecular , Filogenia , Prevalência , Análise de Sequência de DNARESUMO
Infection with human papillomavirus (HPV) is a necessary step in the progression to cervical cancer. Many methods for HPV testing are currently available, mostly developed to detect pools of HPV types. Hybrid Capture 2 (HC2) is one of the most widely used. A new PCR-based assay, the Roche AMPLICOR HPV test, has been recently developed. Both assays recognize a group of 13 HR HPV types contemporaneously. This study evaluated the performance of both methods for detecting high-grade cervical lesions as a part of management for abnormal PAP smears. The study population was composed of 213 women, all referred to colposcopy and histologic diagnosis following an abnormal PAP test. Biopsy-confirmed high-grade cervical intraepithelial neoplasia was used as a gold standard. Overall agreement was 84.9% with a kappa value of 0.6. When comparing the ability to detect moderate cervical intraepithelial neoplasia (CIN2+) and high-grade cervical intraepithelial neoplasia (CIN3+/cancer), AMPLICOR proved slightly more sensitive than HC2, a finding that is important when HPV testing is used in a triage of borderline smear results. Genotyping of discordant results showed a prevalence of LR-HPV types in HC2 positive/AMPLICOR negative samples, and a similar prevalence of HR- and LR-HPV types in AMPLICOR positive/HC2 negative samples. In conclusion, the study shows that the AMPLICOR assay is more sensitive than HC2, which makes it a valid alternative for routine clinical use.
Assuntos
Colo do Útero/virologia , Infecções por Papillomavirus/diagnóstico , Reação em Cadeia da Polimerase/métodos , Displasia do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Adulto , Idoso , Colposcopia , Feminino , Humanos , Pessoa de Meia-Idade , Teste de Papanicolaou , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/virologia , Sensibilidade e Especificidade , Neoplasias do Colo do Útero/virologia , Esfregaço Vaginal , Displasia do Colo do Útero/virologiaRESUMO
Group B streptococcus (GBS) is the most common cause of neonatal and obstetric sepsis and an increasingly important cause of septicemia in elderly subjects and immunocompromised patients. Our aim was to evaluate whether different genotypes of GBS may induce a different production of pro-inflammatory and anti-inflammatory cytokines. We used multilocus sequence typing to identify 71 clones isolated from asymptomatic healthy carriers and symptomatic individuals. All these clinical isolates were used to infect purified human monocytes. TNF-alpha, IL-6, IL-8 and IL-10 secretion was measured. Fifteen allelic sequence types (STs) were identified. The MLST (multilocus sequence typing) analysis grouped the bacteria into four different lineages (clonal cluster) and two of these were closely involved in the infection of symptomatic subjects: CC17 and CC19. Furthermore, CC17 and CC19 stimulated TNF-alpha, IL-6 and IL-8 production significantly more than the other lineages, while CC17 induced a decreased IL-10 production. These results suggest the existence of differences in immune response to infection with particular genotypes of GBS.
Assuntos
Citocinas/metabolismo , DNA Bacteriano , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/microbiologia , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/microbiologia , Streptococcus agalactiae/genética , Idoso , Idoso de 80 Anos ou mais , Citocinas/genética , Feminino , Genótipo , Humanos , Recém-Nascido , Monócitos/imunologia , Monócitos/metabolismo , Monócitos/microbiologia , Gravidez , Complicações Infecciosas na Gravidez/genética , Sepse/microbiologia , Análise de Sequência de DNA , Sorotipagem , Especificidade da Espécie , Infecções Estreptocócicas/genética , Streptococcus agalactiae/classificação , Streptococcus agalactiae/isolamento & purificação , Streptococcus agalactiae/patogenicidade , Virulência/genética , Virulência/imunologiaRESUMO
The prevalence of single and multiple HPV infections was assessed over a cohort of 213 women with cytological abnormalities and its association with cervical neoplasia established. Roche linear array HPV genotyping test was used to identify HPV genotypes. The most prevalent HPV genotypes in cervical cancer samples were HPV16 (61.2%), HPV52 (16.1%), HPV18 (12.9%) and HPV 31 (9.6%). Multiple HR and LR HPV infections, comprising between two and 5+ HPV types, were identified in 49.7% of samples, with a significantly lower number in severe dysplasia and cervical cancer samples (p<0.05). These results seem to indicate that detection of multiple HPV infection with HR-HPV types is not significantly better as a predictor of cervical cancer than single HR-HPV infection, though further longitudinal studies are needed to better clarify the relevance of these infections to the progression of cervical neoplasia.
Assuntos
Infecções por Papillomavirus/epidemiologia , Displasia do Colo do Útero/virologia , Neoplasias do Colo do Útero/virologia , Adulto , Idoso , DNA Viral/análise , Feminino , Humanos , Itália/epidemiologia , Pessoa de Meia-Idade , Papillomaviridae/classificação , Infecções por Papillomavirus/virologia , Prevalência , Esfregaço VaginalRESUMO
MAb IGMB-15, an anti-hIFN-gamma MAb, neutralizes the antiproliferative activity of hIFN-gamma without affecting that of hIFN-alpha or hIFN-beta. The neutralizing capacity of MAb IGMB-15 is wide: it has been assessed on cell lines whose origin and sensitivity to hIFN-gamma differ. The binding of hIFN-gamma to its receptor and its subsequent internalization into the target cell were not influenced by the antibody. MAb IGMB-15 has been found to interact with hIFN-gamma in solution but not when the lymphokine was associated with its cell surface receptor, showing that the growth of certain cell lines can be inhibited at the cell membrane level. This finding is consistent with the existence of an accessory factor responsible for the antiproliferative activity of hIFN-gamma.
Assuntos
Anticorpos Monoclonais/imunologia , Interferon gama/imunologia , Antígenos CD/imunologia , Sítios de Ligação de Anticorpos , Divisão Celular , Reagentes de Ligações Cruzadas , Humanos , Testes de Neutralização , Receptores de Interferon/imunologia , Proteínas Recombinantes , Células Tumorais Cultivadas , Receptor de Interferon gamaRESUMO
In vitro spontaneous apoptosis (SA) of lymphocytes was studied in HIV infection to evaluate possible clinical and prognostic correlations, in a transsectional study of 101 individuals in different clinical categories and in a prospective longitudinal study of 18 asymptomatic individuals (mean follow-up, 17.2 months). The rate of SA was higher in HIV+ patients than in healthy controls (p < 0.001) and was higher in patients with AIDS than in the other HIV+ individuals (p < 0.001). It was inversely correlated with the peripheral blood CD4+ (R -0.61; p < 0.001) and CD8+ (R -0.46; p < 0.001) cell numbers. In a group of long-term survivors (LTS), it was significantly lower than in a control group of asymptomatic HIV+ patients with a similar number of circulating CD4+ lymphocytes but a shorter follow-up (p < 0.02). In the five asymptomatic HIV-infected individuals who showed a clinical progression, peaks of SA rates above the normal range before the clinical event were much more frequent than in those who remained asymptomatic (p < 0.0001), even though they were fairly homogeneous as far as CD4+ cell count and viral load were concerned. The median levels of SA rates were moreover correlated with the rate of total T cell loss (R -0.46; p 0.053). This study suggests that evaluation of the SA levels may provide a predictive factor for clinical and immunological progression of HIV-related immunodeficiencies and strengthen the hypothesis for the role of this phenomenon in the pathogenesis of this progression.
Assuntos
Apoptose , Infecções por HIV/patologia , Adulto , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos/citologia , Estudos Transversais , Feminino , Humanos , Estudos Longitudinais , Depleção Linfocítica , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Sobreviventes , Carga ViralRESUMO
OBJECTIVE: To assess predictive factors of long-term immune restoration in patients who started protease inhibitor (PI)-based HAART and experienced virological rebound after initial complete success. METHOD: A retrospective longitudinal analysis of all HIV-infected patients who started their first PI-based HAART and reached viral load below 500 copies/mL was carried out in a large academic center in Italy. Patients were classified either as complete virologic responder (CR) or rebounders (REB) when confirmed plasma viremia was detected thereafter. Immunological outcome was the area under the curve (AUC) of the absolute CD4+ cell count change since the 8th month after treatment initiation (CD4+ T-cell AUC). Association between baseline characteristics, virological outcome, and CD4+ T-cell AUC was assessed by univariate and multivariate analysis. RESULTS: There were 374 patients who were included in the study. Mean follow-up was 30.2 months. There were 226/374 patients (60.4%) who remained CR while 148/374 (39.6%) presented at least one rebound (REB). Among REB patients, complete viral suppression was regained in 15/42 (35.7%) and 50/106 (47.1%) patients who underwent therapy changes or not, respectively. When multiple linear regression was carried out, previous nucleoside reverse transcriptase inhibitor (NRTI) experience and baseline CD4+ cell count below 350 cells/muL did not impair long-term immune restoration. The occurrence of rebound, its duration (> 18 months), and its magnitude (peak of viral load > 10,000 copies/mL) were independent negative prognostic factors. CONCLUSION: The occurrence of viral rebound is independently associated with significantly impaired long-term immunological restoration. The magnitude of viral rebound (< 10,000 copies/mL) and its duration (< 18 months) may be useful to identify those rebounding patients who may still profit from maintaining the current failing therapy if a more aggressive approach may be expected to be deleterious for tolerability reasons or lack of options.
Assuntos
Terapia Antirretroviral de Alta Atividade , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Inibidores da Protease de HIV/administração & dosagem , Adolescente , Adulto , Idoso , Área Sob a Curva , Contagem de Linfócito CD4 , Esquema de Medicação , Feminino , Infecções por HIV/virologia , HIV-1/genética , HIV-1/imunologia , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , RNA Viral/análise , Estudos Retrospectivos , Resultado do Tratamento , Carga ViralRESUMO
In order to determine whether there is an association between the presence of Epstein-Barr virus (EBV) and mycosis fungoides (MF) disease progression, PCR was performed to detect the EBV status of 20 MF patients; six EBV-positive patients were found. EBV variants may differ in their biological properties, such as their ability to transform cells; therefore, the ability of these variants to immortalize B cells in vitro was analysed. Six continuously growing cell lines were obtained from prolonged cultures of unstimulated peripheral blood mononuclear cells that were taken from the six EBV-positive patients with MF. In order to characterize the EBV strains, EBNA-2 and LMP-1/LMP-2 gene polymorphisms in the six cell lines were also analysed. All patients were followed up for 10 years and it was noticed that EBV-positive patients had a poor prognosis with rapid disease progression and high mortality rates, compared to EBV-negative patients. EBV may therefore constitute a co-factor that accelerates the progression of disease.
Assuntos
Linfócitos B/virologia , Herpesvirus Humano 4/genética , Micose Fungoide/virologia , Polimorfismo Genético , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , Linhagem Celular Transformada , Transformação Celular Viral , Células Cultivadas , Cisteína Endopeptidases/química , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Antígenos Nucleares do Vírus Epstein-Barr/genética , Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Feminino , Herpesvirus Humano 4/classificação , Herpesvirus Humano 4/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Micose Fungoide/fisiopatologia , Reação em Cadeia da Polimerase , Proteínas da Matriz Viral/química , Proteínas da Matriz Viral/genética , Proteínas da Matriz Viral/metabolismo , Proteínas ViraisRESUMO
The aim of this study was to assess the prevalence of genetic changes in either the HIV reverse transcriptase (RT) or protease (Pro) genes in a cohort of patients naïve for anti-retroviral therapy. Of 61 patients, 43 (70.5%) were infected with HIV strains harbouring at least one resistance-related mutation, with 41 (67.2%) harbouring newly recognised treatment-related mutations. Among the 61 patients, the prevalence of specific mutations in the RT gene was as follows: 39A, 1.6%; 43E, 1.6%; and 228H, 1.6%. The prevalence of specific mutations in the Pro gene was as follows: 11I, 1.6%; 13V, 26.2%; 35D, 19.6%; 45R, 1.6%; 58E, 1.6%; 62V, 31%; 72V, 11.4%; 72M, 6.5%; 72T, 3.2%; 75I, 1.6%; and 89M, 13%. A higher prevalence of newly recognised mutations was found in strains from patients infected through sexual practices (30/36 = 83.4% vs. 11/25 = 44%; p 0.0023; OR 10.91; 95% CI 3.14-40.39). These findings support the use of resistance testing in patients naïve for anti-retroviral therapy, and suggest that the possible impact of newly recognised treatment-related mutations on clinical outcome requires further investigation.
Assuntos
Farmacorresistência Viral/genética , Infecções por HIV/epidemiologia , Protease de HIV/genética , Transcriptase Reversa do HIV/genética , HIV-1/efeitos dos fármacos , Mutação , Adulto , Fármacos Anti-HIV/farmacologia , Fármacos Anti-HIV/uso terapêutico , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/virologia , Inibidores da Protease de HIV/farmacologia , Inibidores da Protease de HIV/uso terapêutico , HIV-1/enzimologia , HIV-1/genética , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Inibidores da Transcriptase Reversa/farmacologia , Inibidores da Transcriptase Reversa/uso terapêuticoRESUMO
BACKGROUND: Effectiveness of combination therapy with standard interferon alpha doses and ribavirin is far from being demonstrated in patients with hepatitis C non responders to interferon alpha monotherapy. Recent kinetic studies revealed that these doses may be suboptimal. AIMS: To find the criteria for optimisation of the interferon dose, to be used in combination with ribavirin in patients with hepatitis C non responders to interferon alpha monotherapy. PATIENTS: Sixty-three patients enrolled in a pilot controlled trial were treated for 6 months with ribavirin ([1000-1200 mg daily) and were randomised to concurrently receive interferon alpha 2b for 6 months at: 3 Million Units thrice weekly [group A (21 patients)], 5 MU thrice weekly [group B (21 patients)] and 5 million units daily [group C (21 patients)]. RESULTS: A sustained virological response was observed in: 1 patient from group A (5%), 2 patients from group B (9%) and 8 patients from group C (38%; p=0.02 vs group A; p=0.03 vs group B). Side-effects were not significantly different between the 3 groups. Multivariate analysis showed that infection by hepatitis C virus genotypes 2 or 3 and interferon alpha dosage of 5 million units daily were independent predictors of sustained response. CONCLUSIONS: These results suggest that higher interferon doses administered daily in combination with ribavirin could be more effective in those patients with hepatitis C who had not responded to interferon alone.
Assuntos
Antivirais/administração & dosagem , Hepatite C Crônica/tratamento farmacológico , Interferon-alfa/administração & dosagem , Ribavirina/administração & dosagem , Adolescente , Adulto , Análise de Variância , Biópsia por Agulha , Relação Dose-Resposta a Droga , Esquema de Medicação , Quimioterapia Combinada , Feminino , Seguimentos , Hepatite C Crônica/patologia , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Probabilidade , Índice de Gravidade de Doença , Estatísticas não Paramétricas , Resultado do TratamentoRESUMO
We describe a 65-year-old woman born and living in Northern Italy affected by non tropical spastic tetraparesis and her asymptomatic husband presenting HTLV-1 sequences in their lymphocytes detected by polymerase chain reaction (PCR). We discuss the significance of the case and the mechanism involved in HTLV-1 infection and the relationship with neurological disorders, stressing that this case is the first reported in Italy.
Assuntos
Infecções por HTLV-I/microbiologia , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Linfócitos/microbiologia , Paralisia/microbiologia , Southern Blotting , Western Blotting , Células Cultivadas , Sondas de DNA , DNA Viral/sangue , DNA Viral/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Feminino , Genes env , Genes pX , Genes pol , Anticorpos Anti-HTLV-I/sangue , Antígenos HTLV-I/sangue , Infecções por HTLV-I/diagnóstico , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Paralisia/diagnóstico , Reação em Cadeia da PolimeraseRESUMO
Cells infected with human immunodeficiency virus type 1 (HIV-1) develop viral antigens which can be detected by immunofluorescence. We developed a flow cytometric immunofluorescence assay (FIFA) which provides a quantitative analysis of HIV-1 p24 using a monoclonal antibody (mAb) as a specific reagent. The reduction of HIV p24 antigen expression in viral infected cells was then used to determine HIV neutralizing antibody titers in human sera. Results obtained by FIFA for detecting neutralizing antibodies against HIV-1, when compared with results obtained by indirect immunofluorescence assay (IFA), showed an overall index of agreement of 94.1%. The correlation between the neutralizing antibody titers obtained with each method was found to be highly significant (ro = 0.8; p < 0.01). The simple methodology and the adaptability of this FIFA to various assay conditions make it suitable for routine purposes and for assessing the efficacy of vaccination strategies.
Assuntos
Citometria de Fluxo/métodos , Imunofluorescência , Anticorpos Anti-HIV/sangue , Proteína do Núcleo p24 do HIV/análise , Infecções por HIV/imunologia , HIV-1/imunologia , Anticorpos Monoclonais , Linhagem Celular , Separação Celular , Humanos , Testes de Neutralização , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Mycoplasma have been suggested as co-factors in the pathogenesis of acquired immune deficiency syndrome (AIDS). The prevalence of urethral infection by Mycoplasma genitalium was determined by polymerase chain reaction (PCR) with urethral swabs from 35 HIV-infected patients at different stages of the disease (all of them were heterosexual men). M genitalium was detected in 2 out of 19 non-AIDS (stage A and B) patients and in a similar proportion (1 out of 14; 7.1%) of samples from healthy individuals. A dramatic increase in the frequency of M. genitalium detection was observed in samples of AIDS (stage C) patients. In fact, 9 out of 16 (56.2%) specimens tested positive by PCR. We found no association in AIDS patients between M. genitalium infection and CD4 count, Human Immunodeficiency Virus (HIV) p24 antigenemia or opportunistic infection.