Predictive factors and early biomarkers of response in multiple sclerosis patients treated with natalizumab.

There are an increasing number of treatments available for multiple sclerosis (MS). The early identification of optimal responders to individual treatments is important to achieve individualized therapy. With this aim, we performed a multicenter retrospective longitudinal study including 186 MS patients treated with natalizumab who were followed for 2 years. We analyzed the following variables at recruitment: sex, current age, age at disease onset, disease duration, EDSS, number of T2 and Gd + lesions, IgG and IgM oligoclonal bands, HLA class II (DR, DRB, DQA, DQB, and DRB1*15:01), IgG and IgM antibody titers against human herpesvirus 6 (HHV-6) and the antibody response to Epstein-Barr virus (EBV) through the measurement of the anti-EBNA-1 and anti-VCA IgG titers, in relation to clinical response (no relapses or disability progression), and to NEDA-3 (no evidence of disease activity in terms of clinical response and no changes in MRI scans either) after 2-years follow-up. Baseline EDSS score, baseline EBNA-1 IgG titers and percentage change of HHV6 IgG titers between baseline and 6 month visits were significantly different in clinical responders and in NEDA-3 status (all of them remained significant in the multivariate analysis). We identified three variables for the early identification of natalizumab optimal responders in a rapid and cost-effective approach.

[Epstein Barr virus and invasive mammary carcinomas: EBNA, EBERs and molecular profile in a population of West Algeria]. / Virus d'Epstein-Barr et carcinomes mammaires invasifs : EBNA, EBER et profil moléculaire dans une population de l'ouest algérien.

Breast cancer is the common malignancy that affects women worldwide, but conventional risk factors account for only a small proportion of these cases. A possible viral etiology for breast cancer has been proposed and Epstein-Barr virus (EBV) is a widely studied candidate virus. The objective of this study is to determine the association of EBV infection with infiltrating ductal carcinomas (IDC). This descriptive study was carried out in the laboratory of developmental biology and differentiation, from 2012 to 2014. Of 39 cases, we determined the clinicopathological characteristics of the population. Of the 23 cases of IDC, we implemented the techniques Elisa, immunohistochemistry and in situ hybridization. To determine the serological profile, overexpression of onco-proteins EBNA-1, HER2, the mitotic index Ki67 and detection of the presence of the viral genome. The mean age is 57.40±4, SBR II predominates with 70%, pN+ (27%), RE+ (58%), RP+ (52%), HER2 (81%), Luminal A (34%), Luminal B (14%), HER2 (24%), and triple negative (28%). The serological profile of IgG VCA + in IgG EBNA-1 (87%), EBNA-1 P79 (82%) with a positive relationship between the IgG EBNA-1 and EBNA-1 P79 serology profile (p=0.001), HER2 (p=0.003) and with the molecular profile (p=0.051), EBNA-1 overexpression in (13%). The viral genome (EBER) is found in the tumors 43% representing an inverse relationship with the overexpression of Ki67 and a positive relationship with the overexpression of HER2. In our study we found an association with the presence of the EBV virus and the IDC studied.

The Index of Vulnerability: An anthropological method linking social-ecological systems to mental and physical health outcomes.

Researchers need measures of vulnerability that are grounded in explicit theoretical and conceptual frameworks, that are sensitive to local contexts, and that are easy to collect. This paper presents the Index of Vulnerability (IoV), a quantitative yet anthropologically-informed method connecting social-ecological systems to mental and physical health outcomes. The IoV combines measures of five life domains; food insecurity, water insecurity, access to healthcare, social support, and social status. Scores on this index increase for each life domain where the individual falls into a "high risk" category. Thus, individuals with the highest IoV scores are those who are at risk across multiple life domains. This approach makes the IoV malleable to local contexts, as scholars can choose which measure of each life domain is most appropriate for their study population. An anthropological study conducted among 225 Awajún adults living in the Peruvian Amazon from March to November of 2013 showed that men with higher IoV scores had significantly lower summary fat skinfolds, lower triglyceride levels, and a greater probability of reporting moderate to severe somatic symptoms and poor perceived health. Awajún women with higher IoV scores had significantly elevated perceived stress levels and a greater probability of reporting poor perceived health and moderate to severe somatic and depressive symptoms. Importantly, comparing the IoV to its constituent parts shows that it predicts a wider range of mental and physical health outcomes than any of the life domains alone. The IoV is presented here in relation to the broader political-economic and cultural context of the Awajún, forwarding a critical biocultural approach within anthropology, and demonstrating the IoV's utility for other scholars and practitioners.

Recurrent Natural Killer Cell Lymphoma with Central Nervous System Metastasis Mimicking Cerebellar Infarction.

BACKGROUND: Natural killer cell lymphoma is an uncommon hematologic malignancy, and central nervous system metastasis is rare. The classic magnetic resonance imaging appearance of lymphoma in the brain is T1 hypointense with strong homogeneous gadolinium enhancement, variable T2 signal, and restricted diffusion on diffusion-weighted images. Gadolinium enhancement is an important feature to differentiate lymphoma from infarction. CASE DESCRIPTION: We present the case of a middle-aged man who presented with recurrent natural killer cell lymphoma that metastasized to the cerebellum. Computed tomography and magnetic resonance imaging did not show a contrast-enhancing lesion; imaging features were more suggestive of cerebral infarction. The patient subsequently died, and postmortem examination confirmed natural killer cell lymphoma metastasis to the cerebellum. CONCLUSIONS: Lymphoma can mimic cerebral infarction on computed tomography and magnetic resonance imaging. An imaging appearance of cerebral infarction in a patient with a history of lymphoma should raise suspicions of lymphoma metastasis.

DNA-based hybridization chain reaction for an ultrasensitive cancer marker EBNA-1 electrochemical immunosensor.

An ultrasensitive and selective electrochemical immunosensor was developed for the detection of Epstein Barr virus nuclear antigen 1 (EBNA-1). Firstly, a suspension of graphene sheets (GS) and multi-walled carbon nanotubes (MWCNTs) was prepared with the aid of chitosan (CS) solution and then modified on a glassy carbon electrode (GCE). Gold nanoparticles (AuNPs) were then electrodeposited onto the surface of the GS-MWCNTs film by cyclic voltammetry (CV) to immobilize the captured antibodies. After that, specific sandwich immunoreactions were formed among the captured antibody, EBNA-1, and secondary antibody, DNA-coated carboxyl multi-wall carbon nanotubes (DNA-MWCNTs-Ab2). DNA initiator strands (S0) and secondary antibodies linked to the MWCNTs and double-helix DNA polymers were obtained by hybridization chain reaction (HCR), and here S0 on the MWCNTs propagates a chain reaction of hybridization events between two alternating hairpins to form a nicked double-helix. Finally, electroactive indicator doxorubicin hydrochloride was intercalated into the CG-GC steps between the HCR products and could produce an electrochemical signal, which was monitored by differential pulse voltammetry (DPV). Under optimum conditions, the amperometric signal increased linearly with the target concentrations (0.05-6.4ngmL(-1)), and the immunosensor exhibited a detection limit as low as 0.7pgmL(-1) (S/N=3). The proposed method showed acceptable stability and reproducibility, as well as favorable recovery for EBNA-1 in human serum. The proposed immunosensor provides a novel avenue for signal amplification and potential applications in bioanalysis and clinical diagnostics.

A rare case of undifferentiated nonkeratinizing carcinoma of the lip mucosa.

Undifferentiated nonkeratinizing carcinoma (UNC) is a poorly differentiated squamous cell carcinoma accompanied by a prominent reactive lymphoplasmacytic infiltrate that can occur in many anatomic sites. It shares morphologic features with undifferentiated nonkeratinizing nasopharyngeal carcinoma, in which a strong association with Epstein-Barr virus (EBV) has been noted. Among UNCs arising outside the nasopharynx, the linkage with EBV is variable; in particular, the few cases of UNC of the lip described thus far have been negative for EBV. This report describes a rare case of primary UNC of the lower lip mucosa in a 73-year-old man in whom molecular analysis for EBV showed some amount of viral DNA within the tumor. Surgical excision without adjuvant treatment was performed and the patient was alive without recurrence after 42 months of follow-up. This report presents a rare localization of UNC possibly related to EBV infection and with a good clinical outcome.

Expression of 14-3-3 sigma, cyclin-dependent kinases 2 and 4, p16, and Epstein-Barr nuclear antigen 1 in nasopharyngeal carcinoma.

OBJECTIVE: The protein 14-3-3 sigma plays a role in cell cycle arrest by sequestering cyclin-dependent kinase 1 cyclin B1 complexes, as well as cyclin-dependent kinases 2 and 4, hence its definition as a cyclin-dependent kinase inhibitor. However, the nature of the interaction between these biological markers in nasopharyngeal carcinoma is unknown. This study aimed to investigate whether altered expression of these markers contributes to nasopharyngeal carcinogenesis. METHODS: The study population consisted of 30 nasopharyngeal carcinoma patients and 10 patients without nasopharyngeal carcinoma. The nasopharyngeal carcinoma cell lines TW02, TW04 and Hone-1 were also assessed. We analysed levels of messenger RNA and protein for the p16 gene and the 14-3-3 sigma, Epstein-Barr nuclear antigen 1, and cyclin-dependent kinase 2 and 4 proteins, in nasopharyngeal carcinoma tissue specimens and cell lines and in normal nasopharyngeal tissue. RESULTS: Protein and messenger RNA levels for cyclin-dependent kinase 2 and Epstein-Barr nuclear antigen 1 were significantly higher in nasopharyngeal carcinoma compared with normal tissue, while levels of cyclin-dependent kinase 4 generally were not; results for 14-3-3 sigma varied. Nasopharyngeal carcinoma patients had diminished p16 gene expression, compared with normal tissue. CONCLUSION: Levels of cyclin-dependent kinase 2 and Epstein-Barr nuclear antigen 1 were significantly higher in nasopharyngeal carcinoma than in normal tissue, while p16 gene expression was diminished. These three proteins may contribute to nasopharyngeal carcinogenesis.

[Lymphadenopathy: demarcation to malignant lymphomas]. / Lymphadenopathie: Grenzziehung zu malignen Lymphomen.

Recognition of the differential diagnosis between lymphadenitis and malignant lymphoma requires good knowledge of the basic forms of the disease as well in depth knowledge of the structure of the individual compartments. There are defined forms of lymphadenitis where the differential diagnosis to certain lymphoma entities is known. Other reactive structural alterations show indistinct limits so that a decision is only possible after using additional techniques, such as immunohistochemistry and molecular analyses. Finally, there are marginal areas which can only be clarified by including clinical data.

On optimizing the blocking step of indirect enzyme-linked immunosorbent assay for Epstein-Barr virus serology.

BACKGROUND: Because blocking agent occupies most binding surface of a solid phase, its ability to prevent nonspecific binding determines the signal-to-noise ratio (SNR) and reliability of an enzyme-linked immunosorbent assay (ELISA). METHODS: We demonstrate a stepwise approach to seek a compatible blocking buffer for indirect ELISA, via a case-control study (n=176) of Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC). RESULTS: Regardless of case-control status, we found that synthetic polymer blocking agents, mainly Ficoll and poly(vinyl alcohol) (PVA) were able to provide homogeneous backgrounds among samples, as opposed to commonly used blocking agents, notably nonfat dry milk (NFDM). The SNRs for NPC samples that correspond to blocking using PVA were approximately 3-fold, on average, higher than those blocking using NFDM. Both intra- and inter-assay precisions of PVA-based assays were <14%. CONCLUSION: A blocking agent of choice should have tolerable sample backgrounds from both cases and controls to ensure the reliability of an immunoassay.

Homogeneous assay for detection of active Epstein-Barr nuclear antigen 1 by thrombin activity modulation.

Epstein-Barr virus (EBV) has been associated with several malignancies as Burkitt's lymphoma, nasopharyngeal carcinoma, and Hodgkin's disease. In those diseases, Epstein-Barr nuclear antigen 1 (EBNA-1) is constitutively expressed. Here, we reported an innovative system to detect active EBNA-1 protein in a homogeneous assay. The system is based on the modulation of thrombin activity by a self-complementary single stranded DNA (scssDNA), which was designed and synthesized to mimic the palindromic target sites of EBNA-1 in the EBV genome. This model system showed a limit of detection of 3.75 ng mL(-1) of active EBNA-1 protein with a dynamic detection range from 3.75 to 250 ng mL(-1) with a correlation coefficient of 0.997. This new homogeneous assay for active EBNA-1 protein detection and quantification provides a very useful tool for rapid screening of EBNA-1 blockers in biomedical research.

Effects of long-term use of antiretroviral therapy on the prevalence of oral Epstein-Barr virus.

BACKGROUND: The objectives of this study were to determine (i) the prevalence of oral Epstein-Barr virus (EBV) in HIV-infected subjects compared to non-HIV controls and (ii) the effects of long-term use of antiretroviral therapy (ART) on the prevalence of oral EBV. METHODS: A cross-sectional study was performed in HIV-infected subjects with and without ART, and non-HIV individuals. DNA in saliva samples was extracted and used as a template to detect EBV BamH1W and EBNA1 by quantitative polymerase chain reaction. Student t-test and ANOVA test were performed to determine the prevalence rates among groups. RESULTS: Forty-nine HIV-infected subjects: 37 on ART (age range 23-54 year, mean 37 year), 12 not on ART (age range 20-40 year, mean 31 year), and 20 non-HIV controls (age range 19-53 year, mean 31 year) were enrolled. The numbers of EBV BamH1W in saliva were found to be significantly higher in HIV-infected subjects than non-HIV controls (80% vs. 20%, mean = 12118 vs. 134 copies/10(5) cells, P < 0.001). HIV-infected subjects who were on ART had significantly lower numbers of EBV BamH1W than those who were not (mean = 4102 vs. 138613 copies/10(5) cells, P = 0.011). The numbers were significantly lower in those who received long-term ART compared with short-term (mean = 1401 vs. 11124 copies/10(5) cells, P = 0.034). No significant difference was observed between the groups when using EBNA1 primers. CONCLUSIONS: Prevalence of oral EBV was significantly higher in HIV-infected subjects than non-HIV-controls. The numbers of the virus were significantly decreased by ART. Long-term use of ART did not increase oral EBV.

Label free and amplified detection of cancer marker EBNA-1 by DNA probe based biosensors.

Epstein-Barr virus (EBV) is a human herpes virus that has been associated with several malignancies as Burkitt's lymphoma, nasopharyngeal carcinoma and Hodgkin's disease. All EBV associated malignancies showed a distinct viral gene expression pattern, while Epstein-Barr nuclear antigen 1 (EBNA-1) is constitutively expressed in all such disorders. Here, the development of a biosensor to detect EBNA-1 protein is reported, which was based on a nucleic acid bioreceptor and a quartz crystal microbalance with a dissipation monitoring (QCM-D) transducer. The DNA probe for EBNA-1 detection was designed and synthesized to mimic its palindromic target sites in the EBV genome. This DNA probe was immobilized on the Au-surface of a QCM-D electrode, followed by the blocking of the accessible Au-surface with 6-mercapto-1-hexanol (6-MHO). The system showed a limit of detection of 50 ng/mL in direct detection of EBNA-1, however, the sensitivity was improved by 2 orders of magnitude (0.5 ng/mL) when an amplification cascade, employing antibodies labeled with alkaline phosphatase (AP), was applied to the system.

Localization of Epstein-Barr virus to infiltrating lymphocytes in breast carcinomas and not malignant cells.

The pathogenesis of breast cancer is unknown. In recent years, a number of studies have implicated a role for Epstein-Barr virus (EBV) in a subset of cases. However, these findings are controversial and others have failed to find any link between the virus and this malignancy. We hypothesized that technical differences and the different type and ethnic origin of the cases may be the cause of the disparities reported. Using a highly sensitive EBER-in situ hybridization and immunohistochemistry, we examined 219 samples (158 malignant and 61 non-malignant) from 61 Emirati breast cancer cases to determine if EBV was etiologically associated with Emirati cases and if there was any correlation with other established prognostic factors such as age, histological type, lymph node metastasis, estrogen, progesterone and HER2 expression. We found 47.5% of the cases to be EBV positive, but the virus was localized to occasional infiltrating lymphocytes and not in the malignant cells. EBV lymphocytes were more commonly observed in lymph nodes than in breast tissues, but there was no correlation with malignancy or hormone status. The mean age of our patients was 48years and hormone receptor staining revealed 20% of the cases to be triple negative (ER-/PR-/HER2-). We conclude that although EBV can be detected in breast cancer cases, it is not directly associated with the disease. Thus, a PCR-based approach cannot be used to link this ubiquitous virus to the pathogenesis of breast cancer. Furthermore, we do not find any correlation between the presence of EBV in infiltrating lymphocytes and ER, PR, HER2 expression. We believe our findings will help explain some of the controversies relating to the role of EBV in the pathogenesis of breast cancer.

Geographic variation in the prevalence of Epstein-Barr virus-positive diffuse large B-cell lymphoma of the elderly: a comparative analysis of a Mexican and a German population.

Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma of the elderly was included as a provisional entity in the 2008 WHO lymphoma classification. Most reports of this disease come from Asia and little is known about it in other regions of the world, including Latin America. Therefore, in this study, 305 diffuse large B-cell lymphomas in patients above 50 years were analyzed, 136 from Mexico and 169 from Germany. EBV was detected by Epstein-Barr early RNA (EBER) in situ hybridization. Only cases with EBER+ in the majority of tumor cells were regarded as EBV+ diffuse large B-cell lymphoma. The prevalence of EBV+ diffuse large B-cell lymphoma in Mexican patients was found to be 7% (9 of 136), whereas only 2% (4 of 169) of the German cases were positive. The median age at diagnosis was 66 years in the Mexican cohort, as opposed to 77 years in the German group. The site of presentation was in both groups predominantly nodal in nine cases (70%) and extranodal in four cases (30%). Of the 13 EBV+ cases, 10 (77%) were classified as polymorphic and 3 (23%) as monomorphic type. The polymorphic cases showed a non-germinal center B-cell immunophenotype (CD10- MUM1+). Twelve cases (92%) were LMP1 positive and two (15%) expressed EBNA2. An interesting finding was the high frequency of EBV type B with the LMP1 30 bp deletion found in the Mexican cases (50%). Eight of the 11 evaluable cases were B-cell monoclonal by polymerase chain reaction. In summary, we found a similar prevalence of EBV+ diffuse large B-cell lymphoma of the elderly in a Mexican population compared with what has been reported in Asian countries, and in contrast to the low frequency in Western populations (1-3%). However, compared with the Asian series, the Mexican patients were younger at diagnosis, presented predominantly with nodal disease and rarely expressed EBNA2 protein.

Primary Epstein-Barr virus infection in 2-year-old children: report of 3 cases.

Some children less than four years old have Epstein-Barr virus (EBV)-induced infectious mononucleosis (IM). Because primary EBV infection in infants and young children is usually asymptomatic or subclinical, EBV infection diagnosis may not be easy among young children. To illustrate the clinical characteristics and diagnostic procedures for EBV infection in young children, the authors report herein three cases of primary EBV infection in two-year-old children with an evaluation of their initial clinical symptoms. The results showed that the common initial clinical manifestations are puffy eyelids and hepatosplenomegaly, and that these signs suggest a tentative diagnosis of IM. In conclusion, EBV capsid immunoglobulin (Ig)M antibodies and atypical lymphocytes are useful diagnostic measurements in very young children with symptoms suggestive of IM.

Epidemiological study, immunohistochemistry and in situ hybridization studies of nasopharyngeal carcinomas: a Tunisian report.

Nasopharyngeal carcinomas (NPC) are a significant problem of public health in Tunisia. They are particular because of their characteristic geographic distribution. The aims of this study were, first, to appreciate the presence of Epstein-Barr virus (EBV) genome by immunohistochemistry (IHC) and in situ hybridization (ISH) and to compare their benefits to NPC diagnosis and, secondly, to verify the relation between NPC and factors bound to the food and environment conditions. Biopsies, recruited at the department of pathology of EPS Charles Nicolle at Tunis, were analyzed for EBV genome presence by ISH of EBV-encoded small RNA1 (EBER1). IHC was done with encoded nuclear antigen (EBNA1), latent membrane proteins (LMP1), and antigen BZ1 anti-Z EBV-replication activator (ZEBRA). An epidemiological study based upon the analysis of a detailed questionnaire submitted to patients (all from the north of Tunisia) and 60 witnesses was done. The statistic analysis was realised by SPSS Windows 11.5 Advanced Statistics. All samples were classified as Undifferentiated Carcinoma of Nasopharyngeal type (UCNT). We found a sex ratio of 2 with a bimodal repartition. ISH showed 96.6% positive samples. IHC revealed the EBV in 90% of cases and 66.7%, respectively, with EBNA1 and LMP1. The statistic analysis showed a meaningful relation (P<0.05, OR>3) between NPC and dietary factors (spices and piquant condiment), alcohol and the water quality.

Histological characteristics of 21 Papua New Guinean children with high-grade B-cell lymphoma, which is frequently associated with EBV infection.

The aim of the present study was to confirm the histopathological features of aggressive B-cell lymphoma in Papua New Guinea (PNG)-an EBV endemic region. The immunophenotypic features and expression of EBV-encoded proteins and RNA in B-cell lymphomas were analyzed in 21 PNG children, and compared to the corresponding features of 17 Japanese children with Burkitt lymphoma (BL). Histological diagnosis of the lymphomas from the PNG children was BL in nine patients; atypical Burkitt/Burkitt-like variant of BL (BLL) in three; diffuse large B-cell lymphoma (DLBCL) in four; and B-lymphoblastic lymphoma (B-LBL) in five. The lymphomas from the PNG children had a high positive rate on EBV-RNA in situ hybridization (EBV-ISH; 66.7%). With regard to the histological typing, 10 of 12 patients (83%) with BL/BLL, one of four (25%) with DLBCL, and three of five (60%) with B-LBL were positive for EBV-ISH. The findings of EBV-positive B-LBL were surprising because it is commonly considered that lymphoblastic lymphoma is not associated with EBV. EBV positivity was not detected in the 12 Japanese patients who were available for the EBV-ISH evaluation. It is concluded that it is possible that a proportion of DLBCL and B-LBL besides BL/BLL are associated with EBV in endemic region.

EBV-associated diffuse large B-cell lymphoma arising in the chest wall with surgical mesh implant.

In Japan, most cases of malignant lymphoma arising in the thorax are pyothorax-associated lymphoma, which develops in patients who have undergone artificial pneumothorax, used in the past as surgical therapy for pulmonary tuberculosis. Pyothorax-associated lymphoma consist mostly of diffuse large B-cell lymphoma and have a strong association with EBV. Herein is reported the case of a diffuse large B-cell lymphoma arising in the left thoracic wall after left lung resection for squamous cell carcinoma and chest wall reconstruction with polyethylene terephthalate (PET) surgical mesh. The tumor was found 20 years after the operation and was confined to the chest wall adjacent to the PET mesh. The patient did not have a clinical history of pyothorax after surgery. The lymphoma cells were of the large cell type and were positive for CD20, EBV-encoded small RNA--in situ hybridization, LMP-1 and EBNA-2. The present case demonstrates that EBV-related B-cell lymphoma can occur after surgery other than artificial pneumothorax. In the present case, long-standing chronic inflammation induced by PET mesh may have been associated with the development of lymphoma.