Comparison of immunofluorescence with enzyme immunoassay for detection of Q fever

To evaluate enzyme immunoassay (EIA) as an alternative to indirect immunofluorescence assay (IFA) to screen for Q fever in humans, 157 serum samples from patients suspected of having the disease were tested for immunoglobulin G antibodies toCoxiella burnetii. The agreement between the tests and the sensitivity of EIA were excellent (96.8% and 98.4%, respectively) when an IFAtiter of > 1/160 was considered positive. All serum samples with a titer of > 1/320 in the IFA were also positive by the EIA. The EIA seems to be an acceptable alternative to IFA for screening for Q fever.     

Generation of a Novel In Vitro Model to Study Endothelial Dysfunction from Atherothrombotic Specimens

Cardiovascular Drugs and Therapy - Endothelial dysfunction is central to the pathogenesis of acute coronary syndrome. The study of diseased endothelium is very challenging due to inherent...     

Evaluación de la utilización de una consulta de diagnóstico rápido de cáncer de pulmón. Tiempos de demora diagnóstica y terapéutica

Objective

To analyze the results obtained in a lung cancer screening program since its inception five years ago regarding correct referrals, diagnostic and therapeutic delay times and days of hospitalization. To compare the diagnostic-therapeutic delays and hospital stays with those obtained in patients evaluated with the standard system.

Patients and methods

Included for study were all those patients evaluated in our Lung Cancer Screening Program (LCSP) in the last five years. For the cases with LC, we recorded the dates the patients were referred to a specialist, the first consultation, diagnostic tests, stage, start of treatment and days of hospitalization. We compared these same data with lung cancer patients who did not partake in the LCSP and were diagnosed between October 2008 and October 2010.

Results

We evaluated 179 patients remitted to the LCSP, which represented 26.7% of the consultations; 166 (92.7%) of the referrals were correct, out of which 44.5% were LC. In 75.6% of these, the entire study was completed in the outpatient setting, and more than 85% of the cases met the current recommendations related with diagnostic-therapeutic delays. When these results were compared with the non-LCSP group (n = 151), differences were found in the data for hospitalizations: there was a lower percentage of hospitalizations (P < .0001) and shorter hospital stays (P < .0001) in the LCSP group. There were no differences between the two groups for diagnostic or therapeutic delays.

Conclusion

In our setting, lung cancer screening programs allow for cancer studies to be carried out in the outpatient consultations in a large percentage of cases, and within the time periods recommended by current guidelines. In spite of this fact, we have detected that these programs are underused.     

Systems Medicine: from molecular features and models to the clinic in COPD

Background and hypothesis

Chronic Obstructive Pulmonary Disease (COPD) patients are characterized by heterogeneous clinical manifestations and patterns of disease progression. Two major factors that can be used to identify COPD subtypes are muscle dysfunction/wasting and co-morbidity patterns. We hypothesized that COPD heterogeneity is in part the result of complex interactions between several genes and pathways. We explored the possibility of using a Systems Medicine approach to identify such pathways, as well as to generate predictive computational models that may be used in clinic practice.

Objective and method

Our overarching goal is to generate clinically applicable predictive models that characterize COPD heterogeneity through a Systems Medicine approach. To this end we have developed a general framework, consisting of three steps/objectives: (1) feature identification, (2) model generation and statistical validation, and (3) application and validation of the predictive models in the clinical scenario. We used muscle dysfunction and co-morbidity as test cases for this framework.

Results

In the study of muscle wasting we identified relevant features (genes) by a network analysis and generated predictive models that integrate mechanistic and probabilistic models. This allowed us to characterize muscle wasting as a general de-regulation of pathway interactions. In the co-morbidity analysis we identified relevant features (genes/pathways) by the integration of gene-disease and disease-disease associations. We further present a detailed characterization of co-morbidities in COPD patients that was implemented into a predictive model. In both use cases we were able to achieve predictive modeling but we also identified several key challenges, the most pressing being the validation and implementation into actual clinical practice.

Conclusions

The results confirm the potential of the Systems Medicine approach to study complex diseases and generate clinically relevant predictive models. Our study also highlights important obstacles and bottlenecks for such approaches (e.g. data availability and normalization of frameworks among others) and suggests specific proposals to overcome them.

     

Primary retinal vasculitis

CASE REPORT: We present the case of a 22-year-old Caucasian female who consulted because of a brief episode of blurred vision in her right eye. Funduscopic findings in right eye included marked venous tortuousness and roundish paramacular haemorrhages. A slight venous tortuousness in left eye. Fluorescein angiography showed a segmentary retinal vasculitis affecting medium calibre vessels of the posterior pole. Complementary examinations ruled out any systemic involvement. After therapy with oral corticoesteroids the case evolves to remission. DISCUSSION: The diagnosis of retinal vasculitis is achieved by the performance of ophthalmoscopy and fluorescein angiography. Prior studies have ruled out the need for complementary examinations to find any systemic involvement in patients.     

Metabolic sensitivity of pancreatic tumour cell apoptosis to glycogen phosphorylase inhibitor treatment

Inhibitors of glycogen breakdown regulate glucose homeostasis by limiting glucose production in diabetes. Here we demonstrate that restrained glycogen breakdown also inhibits cancer cell proliferation and induces apoptosis through limiting glucose oxidation, as well as nucleic acid and de novo fatty acid synthesis. Increasing doses (50-100 microM) of the glycogen phosphorylase inhibitor CP-320626 inhibited [1,2-(13)C(2)]glucose stable isotope substrate re-distribution among glycolysis, pentose and de novo fatty acid synthesis in MIA pancreatic adenocarcinoma cells. Limited oxidative pentose-phosphate synthesis, glucose contribution to acetyl CoA and de novo fatty acid synthesis closely correlated with decreased cell proliferation. The stable isotope-based dynamic metabolic profile of MIA cells indicated a significant dose-dependent decrease in macromolecule synthesis, which was detected at lower drug doses and before the appearance of apoptosis markers. Normal fibroblasts (CRL-1501) did not show morphological or metabolic signs of apoptosis likely due to their slow rate of growth and metabolic activity. This indicates that limiting carbon re-cycling and rapid substrate mobilisation from glycogen may be an effective and selective target site for new drug development in rapidly dividing cancer cells. In conclusion, pancreatic cancer cell growth arrest and death are closely associated with a characteristic decrease in glycogen breakdown and glucose carbon re-distribution towards RNA/DNA and fatty acids during CP-320626 treatment.     

GCV modulates the antitumoural efficacy of a replicative adenovirus expressing the Tat8-TK as a late gene in a pancreatic tumour model

Replication-competent adenoviruses carrying the herpes simplex thymidine kinase (TK) gene have shown contradictory evidence with regard to their antitumoural efficacy in combination with ganciclovir (GCV) treatment. We generated a replication-competent adenovirus carrying Tat8-TK, a modified form of the TK gene, under the control of the adenoviral major late promoter (AdRGDTat8-TK-L). Pancreatic cancer cell lines with different sensitivity to the TK/GCV system were infected with AdRGDTat8-TK-L, both in the presence and absence of GCV, and tested for treatment efficacy. We observed that, although the presence of GCV reduced viral replication in all infected cell lines, in three out of four GCV significantly enhanced the efficacy of the virotherapy. Interestingly, the cytotoxicity of the AdRGD-Tat8-TK-L/GCV was found more potent than that of a first generation AdTK/GCV system. In tumour xenografts from BxPC-3 and NP-18 pancreatic cells, both AdRGDTat8-TK-L and AdRGDTat8-TK-L/GCV treatment showed antitumoural activity. In BxPC-3 tumours scheduling of virus and prodrug was a key factor to determine the outcome of the therapy. Importantly, the addition of GCV enhanced the antitumoural effect of AdRGDTat8-TK-L only when applied in two rounds of virus+GCV. Interestingly, in spite of interfering with viral replication in vitro, GCV treatment of NP-18 tumours did not compromise the antitumoural efficacy of the AdRGDTat8-TK-L adenovirus. Thus, our results show that the combination therapy of a replicative adenovirus and the Tat8-TK/GCV suicide system can prove beneficial, when the appropriate regimen of virus and GCV is applied.