CCR2 and CCR5 genes polymorphisms in women with cervical lesions from Pernambuco, Northeast Region of Brazil: a case-control study

Polymorphisms in chemokine receptors play an important role in the progression of cervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examined the association of CCR2-64I (rs1799864) andCCR5-Δ32 (rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) in a Brazilian population. The genotyping of 139 women with cervical lesions and 151 women without cervical lesions for the CCR2-64I and CCR5-Δ32 polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism. The individuals carrying heterozygous or homozygous genotypes (GA+AA) for CCR2-64I polymorphisms seem to be at lower risk for cervical lesion [odds ratio (OR) = 0.37, p = 0.0008)]. The same was observed for the A allele (OR = 0.39, p = 0.0002), while no association was detected (p > 0.05) with CCR5-Δ32 polymorphism. Regarding the human papillomavirus (HPV) type, patients carrying the CCR2-64Ipolymorphism were protected against infection by HPV type 16 (OR = 0.35, p = 0.0184). In summary, our study showed a protective effect ofCCR2-64I rs1799864 polymorphism against the development of cervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.

The role of polymerase chain reaction of high-risk human papilloma virus in the screening of high-grade squamous intraepithelial lesions in the anal mucosa of human immunodeficiency virus-positive males having sex with males.

OBJECTIVES To evaluate the advantages of cytology and PCR of high-risk human papilloma virus (PCR HR-HPV) infection in biopsy-derived diagnosis of high-grade squamous intraepithelial lesions (HSIL = AIN2/AIN3) in HIV-positive men having sex with men (MSM). METHODS This is a single-centered study conducted between May 2010 and May 2014 in patients (n = 201, mean age 37 years) recruited from our outpatient clinic. Samples of anal canal mucosa were taken into liquid medium for PCR HPV analysis and for cytology. Anoscopy was performed for histology evaluation. RESULTS Anoscopy showed 33.8% were normal, 47.8% low-grade squamous intraepithelial lesions (LSIL), and 18.4% HSIL; 80.2% had HR-HPV. PCR of HR-HPV had greater sensitivity than did cytology (88.8% vs. 75.7%) in HSIL screening, with similar positive (PPV) and negative predictive value (NPV) of 20.3 vs. 22.9 and 89.7 vs. 88.1, respectively. Combining both tests increased the sensitivity and NPV of HSIL diagnosis to 100%. Correlation of cytology vs. histology was, generally, very low and PCR of HR-HPV vs. histology was non-existent (<0.2) or low (<0.4). Area under the receiver operating characteristics (AUROC) curve analysis of cytology and PCR HR-HPV for the diagnosis of HSIL was poor (<0.6). Multivariate regression analysis showed protective factors against HSIL were: viral suppression (OR: 0.312; 95%CI: 0.099-0.984), and/or syphilis infection (OR: 0.193; 95%CI: 0.045-0.827). HSIL risk was associated with HPV-68 genotype (OR: 20.1; 95%CI: 2.04-197.82). CONCLUSIONS When cytology and PCR HR-HPV findings are normal, the diagnosis of pre-malignant HSIL can be reliably ruled-out in HIV-positive patients. HPV suppression with treatment protects against the appearance of HSIL.

The microbiological signature of human cutaneous leishmaniasis lesions exhibits restricted bacterial diversity compared to healthy skin

Localised cutaneous leishmaniasis (LCL) is the most common form of cutaneous leishmaniasis characterised by single or multiple painless chronic ulcers, which commonly presents with secondary bacterial infection. Previous culture-based studies have found staphylococci, streptococci, and opportunistic pathogenic bacteria in LCL lesions, but there have been no comparisons to normal skin. In addition, this approach has strong bias for determining bacterial composition. The present study tested the hypothesis that bacterial communities in LCL lesions differ from those found on healthy skin (HS). Using a high throughput amplicon sequencing approach, which allows for better populational evaluation due to greater depth coverage and the Quantitative Insights Into Microbial Ecology pipeline, we compared the microbiological signature of LCL lesions with that of contralateral HS from the same individuals.Streptococcus, Staphylococcus,Fusobacterium and other strict or facultative anaerobic bacteria composed the LCL microbiome. Aerobic and facultative anaerobic bacteria found in HS, including environmental bacteria, were significantly decreased in LCL lesions (p < 0.01). This paper presents the first comprehensive microbiome identification from LCL lesions with next generation sequence methodology and shows a marked reduction of bacterial diversity in the lesions.

Cynomolgus monkeys (Macaca fascicularis) experimentally infected with B19V and hepatitis A virus: no evidence of the co-infection as a cause of acute liver failure

This study was conducted to analyse the course and the outcome of the liver disease in the co-infected animals in order to evaluate a possible synergic effect of human parvovirus B19 (B19V) and hepatitis A virus (HAV) co-infection. Nine adult cynomolgus monkeys were inoculated with serum obtained from a fatal case of B19V infection and/or a faecal suspension of acute HAV. The presence of specific antibodies to HAV and B19V, liver enzyme levels, viraemia, haematological changes, and necroinflammatory liver lesions were used for monitoring the infections. Seroconversion was confirmed in all infected groups. A similar pattern of B19V infection to human disease was observed, which was characterised by high and persistent viraemia in association with reticulocytopenia and mild to moderate anaemia during the period of investigation (59 days). Additionally, the intranuclear inclusion bodies were observed in pro-erythroblast cell from an infected cynomolgus and B19V Ag in hepatocytes. The erythroid hypoplasia and decrease in lymphocyte counts were more evident in the co-infected group. The present results demonstrated, for the first time, the susceptibility of cynomolgus to B19V infection, but it did not show a worsening of liver histopathology in the co-infected group.

Assessment of melanocytic skin lesions with a high-definition laser Doppler imaging system.

BACKGROUND: Early detection is a major goal in the management of malignant melanoma. Besides clinical assessment many noninvasive technologies such as dermoscopy, digital dermoscopy and in vivo laser scanner microscopy are used as additional methods. Herein we tested a system to assess lesional perfusion as a tool for early melanoma detection.¦METHODS: Laser Doppler flow (FluxExplorer) and mole analyser (MA) score (FotoFinder) were applied to histologically verified melanocytic nevi (33) and malignant melanomas (12).¦RESULTS: Mean perfusion and MA scores were significantly increased in melanoma compared to nevi. However, applying an empirically determined threshold of 16% perfusion increase only 42% of the melanomas fulfilled the criterion of malignancy, whereas with the mole analyzer score 82% of the melanomas fulfilled the criterion of malignancy.¦CONCLUSION: Laser Doppler imaging is a highly sensitive technology to assess skin and skin tumor perfusion in vivo. Although mean perfusion is higher in melanomas compared to nevi the high numbers of false negative results hamper the use of this technology for early melanoma detection.

Dopamine reverses reward insensitivity in apathy following globus pallidus lesions.

Apathy is a complex, behavioural disorder associated with reduced spontaneous initiation of actions. Although present in mild forms in some healthy people, it is a pathological state in conditions such as Alzheimer's and Parkinson's disease where it can have profoundly devastating effects. Understanding the mechanisms underlying apathy is therefore of urgent concern but this has proven difficult because widespread brain changes in neurodegenerative diseases make interpretation difficult and there is no good animal model. Here we present a very rare case with profound apathy following bilateral, focal lesions of the basal ganglia, with globus pallidus regions that connect with orbitofrontal (OFC) and ventromedial prefrontal cortex (VMPFC) particularly affected. Using two measures of oculomotor decision-making we show that apathy in this individual was associated with reward insensitivity. However, reward sensitivity could be established partially with levodopa and more effectively with a dopamine receptor agonist. Concomitantly, there was an improvement in the patient's clinical state, with reduced apathy, greater motivation and increased social interactions. These findings provide a model system to study a key neuropsychiatric disorder. They demonstrate that reward insensitivity associated with basal ganglia dysfunction might be an important component of apathy that can be reversed by dopaminergic modulation.

Novel immunotherapeutic strategies against human papillomavirus type 16 (HPV 16) induced lesions and cervical cancer

RESUME Le cancer du col de l'utérus, deuxième cause de mort par cancer chez la femme, a pu être associé à une infection par plusieurs types de virus du Papillome Humain (HPV), et en particulier HPV 16. Les vaccins prophylactiques sont efficaces à prévenir le cancer du col utérin alors que les lésions de haut grade sont généralement traitées par ablation chirurgicale et par d'éventuels traitements additionnels. Les risques de récurrence liés aux ablations et le taux de mortalité (50%) lié au cancer, démontrent le besoin de développer des stratégies alternatives afin de cibler les lésions précancéreuses. A ce jour, les vaccins thérapeutiques ont démontré peu de résultats cliniques, contrastant avec les régressions de tumeurs ectopiques observées après vaccination dans des modèles murins avec tumeurs associées à HPV. L'induction de réponses immunitaires protectrices dans la muqueuse génitale semble être cruciale pour l'efficacité des vaccins thérapeutiques HPV et évaluer leur efficacité dans un modèle murin avec tumeurs-HPV génitales représente un pré-requis important avant de procéder à des études cliniques. Par conséquent, nous avons établi un modèle murin orthotopique où des tumeurs se développent dans (a muqueuse génitale après une instillation intra-vaginale (i.vag) de cellules tumorales exprimant les oncogènes E6/E7 d'HPV 16 et transduites par un vecteur lentiviral codant la luciferase afin de suivre le développement de ces tumeurs in vivo par imagerie. La caractérisation histologique a démontré que les tumeurs grandissaient dans l'épithélium vaginal et en accord avec leur localisation, des cellules Τ CD8 spécifiques à E7 induites par la tumeur n'étaient détectées que dans la muqueuse génitale et les ganglions drainants. Une infiltration de cellules Τ régulatrices a aussi été mise en évidence, empêchant la régression spontanée de ces tumeurs. Par conséquent, ce modèle devrait être plus adéquat pour tester des stratégies thérapeutiques, étant donné qu'il partage certaines similarités immunologiques avec les lésions génitales naturelles causées par HPV. Etant donné que les oncogènes E6 et E7 d'HPV sont nécessaires à la maintenance du phénotype cancéreux des cellules cervicales, elles représentent des antigènes cibles pour la vaccination thérapeutique. Nous avons démontré que des souris immunisées par voie sous-cutanée (s.c.) avec une dose d'un vaccin à base de polypeptide E7 d'HPV 16 et d'adjuvants, présentaient de nombreuses cellules Τ CD8 sécrétant de l'IFN-γ spécifiquement à E7 dans leurs organes lymphatiques mais également dans la muqueuse génitale. De plus, le manque de corrélation entre les réponses spécifiques mesurées dans la périphérie et dans la muqueuse génitale souligne la nécessité et l'importance de déterminer les réponses immunitaires localement là où les lésions dues à HPV se développent. Si une vaccination par voie muqueuse est plus propice à traiter/régresser des infections génitales/tumeurs que le voie parentérale est un sujet débattu. Nos données montrent que seule la voie s.c. était capable de régresser la quasi totalité des tumeurs génitales chez la souris bien que des réponses CD8 spécifiques à E7 similaires étaient mesurées dans la muqueuse génitale après des vaccinations intra-nasale et i.vag. Afin d'augmenter la réponse spécifique au vaccin dans la muqueuse génitale, des immunostimulants ont été administrés par voie i.vag après vaccination. Nous avons démontré qu'une application i.vag d'agonistes des Toll like receptors après une vaccination s.c. induisait de manière significative une augmentation des cellules Τ CD8 sécrétant de l'IFN-γ spécifiquement à E7 dans la muqueuse génitale. Plus précisément et concernant les CpG et Poly l:C, l'effet était probablement associé à une attraction locale des cellules Τ CD8 et deuxièmement dépendait respectivement des voies de signalisation TLR9 et TLR3/Mda5. Finalement, cette stratégie combinatoire a permis de régresser des grosses tumeurs génitales chez la souris, suggérant qu'une telle immunothérapie pourrait adéquatement traiter des lésions dues à HPV chez les femmes. SUMMARY Cervical cancer is the second leading cause of cancer mortality in women worldwide and results from an infection with a subset of Human Papillomavirus (HPV), HPV 16 representing the most prevalent type. The available prophylactic vaccines are an effective strategy to prevent cervical cancer while already established high grade lesions usually require surgical ablation of lesion with possible additional treatments. Recurrence risks linked to conventional ablations and the high mortality (50%) related to cervical cancer demonstrate the need for alternative strategies like immunotherapies to target pre¬cancerous lesions. Until now, therapeutic vaccines only showed limited clinical results, which strongly contrast with the regression of ectopic tumors observed in the available murine HPV tumor models after vaccination. Induction of protective immune responses in the genital mucosa (GM) may be crucial for efficacy of HPV therapeutic vaccines and evaluating their efficacy in a murine model with genital HPV- tumors represents an important prerequisite for clinical trials. Thus, we have here established an orthotopic mouse model where tumors in the GM develop after an intravaginal (i.vag) instillation of HPV 16 E6/E7 oncogenes-expressing tumor cells transduced with a luciferase encoding lentivirus vector for in vivo imaging of tumor growth. Histological characterization showed that tumor grew within the vaginal epithelium and according to their mucosal location tumor- induced E7-specific CD8 Τ cells were restricted to the GM and genital draining lymph nodes together with high Τ regulatory cells infiltrates preventing spontaneous regression. Consequently, sharing several immunological similarities with natural genital HPV lesions, this novel genital tumor model may be more adequate to test therapeutic strategies. As E6 and/or E7 HPV oncogenes expression is required for the maintenance of the cancerous phenotype of cervical cells, they represent target antigens for therapeutic vaccination. We reported that mice subcutaneously (s.c.) immunized once with an adjuvanted HPV 16 E7 polypeptide vaccine harbored high E7-specific IFN-γ secreting CD8 Τ cells in their lymphoid organs and more importantly in the GM. In addition, the lack of correlation between specific responses measured in the periphery with those measured in the GM highlighted the necessity and relevance to determine the immune responses locally where HPV 16-induced lesions develop. Whether a mucosal route of immunization is better to treat/regress genital infections/tumors than parenteral immunization is still debated. Our data shows that although similar E7-specific IFN-γ secreting CD8 Τ cells responses were measured in the GM upon mucosal routes of E7 vaccine delivery (nasal and vaginal immunizations), only the s.c immunization was able to regress at least all genital tumors in mice. To further increase the vaccine-specific responses in the GM, immunostimulatory agents were i.vag administrated after vaccination. We demonstrated that a single i.vag application of toll like receptor (TLR) agonists after a s.c. E7 vaccination induced a significant increase of E7-specific IFN-γ secreting CD8 Τ cells in the GM. More precisely, regarding CpG and Poly l:C, the effect is most probably associated with a local attraction of total CD8 Τ cells and secondly depends on TLR9 and TLR3/Mda5 signaling pathways, respectively. Finally, this combinatorial strategy induced tumor regression in mice harboring large genital tumors, suggesting that such an immunotherapy could be adequate to treat HPV-induced lesions in women.

Diagnostic Accuracy of (18)F-FDG-PET and PET/CT in the Differential Diagnosis between Malignant and Benign Pleural Lesions: A Systematic Review and Meta-Analysis.

RATIONALE AND OBJECTIVES: To systematically review and meta-analyze published data about the diagnostic accuracy of fluorine-18-fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET) and PET/computed tomography (CT) in the differential diagnosis between malignant and benign pleural lesions. METHODS AND MATERIALS: A comprehensive literature search of studies published through June 2013 regarding the diagnostic performance of (18)F-FDG-PET and PET/CT in the differential diagnosis of pleural lesions was carried out. All retrieved studies were reviewed and qualitatively analyzed. Pooled sensitivity, specificity, positive and negative likelihood ratio (LR+ and LR-) and diagnostic odds ratio (DOR) of (18)F-FDG-PET or PET/CT in the differential diagnosis of pleural lesions on a per-patient-based analysis were calculated. The area under the summary receiver operating characteristic curve (AUC) was calculated to measure the accuracy of these methods. Subanalyses considering device used (PET or PET/CT) were performed. RESULTS: Sixteen studies including 745 patients were included in the systematic review. The meta-analysis of 11 selected studies provided the following results: sensitivity 95% (95% confidence interval [95%CI]: 92-97%), specificity 82% (95%CI: 76-88%), LR+ 5.3 (95%CI: 2.4-11.8), LR- 0.09 (95%CI: 0.05-0.14), DOR 74 (95%CI: 34-161). The AUC was 0.95. No significant improvement of the diagnostic accuracy considering PET/CT studies only was found. CONCLUSIONS: (18)F-FDG-PET and PET/CT demonstrated to be accurate diagnostic imaging methods in the differential diagnosis between malignant and benign pleural lesions; nevertheless, possible sources of false-negative and false-positive results should be kept in mind.

Selective accumulation of differentiated FOXP3(+) CD4 (+) T cells in metastatic tumor lesions from melanoma patients compared to peripheral blood

Precise identification of regulatory T cells is crucial in the understanding of their role in human cancers. Here, we analyzed the frequency and phenotype of regulatory T cells (Tregs), in both healthy donors and melanoma patients, based on the expression of the transcription factor FOXP3, which, to date, is the most reliable marker for Tregs, at least in mice. We observed that FOXP3 expression is not confined to human CD25(+/high) CD4(+) T cells, and that these cells are not homogenously FOXP3(+). The circulating relative levels of FOXP3(+) CD4(+) T cells may fluctuate close to 2-fold over a short period of observation and are significantly higher in women than in men. Further, we showed that FOXP3(+) CD4(+) T cells are over-represented in peripheral blood of melanoma patients, as compared to healthy donors, and that they are even more enriched in tumor-infiltrated lymph nodes and at tumor sites, but not in normal lymph nodes. Interestingly, in melanoma patients, a significantly higher proportion of functional, antigen-experienced FOXP3(+) CD4(+) T was observed at tumor sites, compared to peripheral blood. Together, our data suggest that local accumulation and differentiation of Tregs is, at least in part, tumor-driven, and illustrate a reliable combination of markers for their monitoring in various clinical settings.

Fissures du labrum glénoïdal postérieur et lésions associées de l'épaule: une étude en arthroscanner [Fissures of the posterior labrum and associated lesions: CT arthrogram evaluation]

PURPOSE: To describe osseous, chondral and tendinous lesions associated with fissures of the posterior labrum. To better understand the pathological processes leading to fissure of the posterior labrum. Materials and methods. Retrospective study of 43 CT arthrograms performed in 43 patients that showed a fissure of the posterior labrum. The following associated lesions were noted: osseous and chondral remodeling of the humeral head and/or glenoid and articular surface fissures of the rotator cuff. Based on type of associated lesions, patients were separated into one of four pathological subgroups: posterior instability, posterosuperior or internal impingement, anterior instability and isolated fissure of the posterior labrum. RESULTS: Sixteen patients (37.2%) of patients showed posterior instability, 12 (27.9%) showed lesions of internal impingement, and 11 (25.6%) showed lesions of anterior instability. Only 4 patients (9.3%) had an isolated fissure of the posterior labrum. CONCLUSION: Posterior instability, internal impingement and anterior instability are the main pathologies leading to fissure of the posterior labrum, which seldom occurs in isolation. Evaluation of these associated lesions allows understanding of the underlying pathological processes leading to fissure of the posterior labrum.

Interobserver and intraobserver variability of the apparent diffusion coefficient in treated malignant hepatic lesions on a 3.0T machine: measurements in the whole lesion versus in the area with the most restricted diffusion.

PURPOSE: To assess the inter/intraobserver variability of apparent diffusion coefficient (ADC) measurements in treated hepatic lesions and to compare ADC measurements in the whole lesion and in the area with the most restricted diffusion (MRDA). MATERIALS AND METHODS: Twenty-five patients with treated malignant liver lesions were examined on a 3.0T machine. After agreeing on the best ADC image, two readers independently measured the ADC values in the whole lesion and in the MRDA. These measurements were repeated 1 month later. The Bland-Altman method, Spearman correlation coefficients, and the Wilcoxon signed-rank test were used to evaluate the measurements. RESULTS: Interobserver variability for ADC measurements in the whole lesion and in the MRDA was 0.17 x 10(-3) mm(2)/s [-0.17, +0.17] and 0.43 x 10(-3) mm(2)/s [-0.45, +0.41], respectively. Intraobserver limits of agreement could be as low as [-0.10, +0.12] 10(-3) mm(2)/s and [-0.20, +0.33] 10(-3) mm(2)/s for measurements in the whole lesion and in the MRDA, respectively. CONCLUSION: A limited variability in ADC measurements does exist, and it should be considered when interpreting ADC values of hepatic malignancies. This is especially true for the measurements of the minimal ADC.

Immunoscintigraphic localization of inflammatory lesions: concept, radiolabelling and in vitro testing of a granulocyte specific antibody.

Current nuclear medicine techniques for the localization of inflammatory processes are based on injection of 111In labelled autologous granulocytes which need to be isolated and radiolabelled in vitro before reinjection. A new technique is presented here that obviates the need for cell isolation by the direct intravenous injection of a granulocyte specific 123I labelled monoclonal antibody. In this publication the basic parameters of the antibody granulocyte interaction are described. Antibody binding does not inhibit vital functions of the granulocytes, such as chemotaxis and superoxide generation. Scatchard analysis of binding data reveals an apparent affinity of the antibody for granulocytes of 6.8 X 10(9) l/mol and approximately 7.1 X 10(4) binding sites per cell. Due to the high specificity of the antibody, the only expected interference is from CEA producing tumors.