Feasibility of whole genome and transcriptome profiling in pediatric and young adult cancers.

The utility of cancer whole genome and transcriptome sequencing (cWGTS) in oncology is increasingly recognized. However, implementation of cWGTS is challenged by the need to deliver results within clinically relevant timeframes, concerns about assay sensitivity, reporting and prioritization of findings. In a prospective research study we develop a workflow that reports comprehensive cWGTS results in 9 days. Comparison of cWGTS to diagnostic panel assays demonstrates the potential of cWGTS to capture all clinically reported mutations with comparable sensitivity in a single workflow. Benchmarking identifies a minimum of 80× as optimal depth for clinical WGS sequencing. Integration of germline, somatic DNA and RNA-seq data enable data-driven variant prioritization and reporting, with oncogenic findings reported in 54% more patients than standard of care. These results establish key technical considerations for the implementation of cWGTS as an integrated test in clinical oncology.

Menopausia: actitudes y conocimientos en un grupo de medicos internos, de la facultad de medicina, de la Universidad de Antioquia / Menopause: attiude an knowledge in a group of final-year medical students of the school of medicine of the University of Antioquia

Se presentan los resultados de una encuesta realizada a los médicos de la Facultad de Medicina de la Universidad de Antioquia, en el segundo semestre de 1997. El proyecto se realizó con el doble propósito de construir conocimiento sobre el tema del climaterio femenino y, en segundo lugar, para aportar elementos pertinentes a la discusión de cambio curricular en que está empeñada la Facultad y la Universidad. Partimos de la hipótesis de que la historia personal del individuo limita las actitudes y condiciona el desarrollo de motivación y destrezas sobre un tema determinado. Se utilizó el censo y las técnicas de encuesta y escala Lickert para indagar sobre variables socioculturales, demográficas y de carácter científico/técnico. Se encontró que el nivel de conocimientos, medido por la nota, fue de 2.85 + 0.83; la actitud frente a la terapia fue baja en el 44 por ciento y alta en el 40 por ciento. Como se había previsto en la hipótesis, la relación entre actitudes y conocimientos fue directa, pero la asociación con variables sociodemográficas no pudo determinarse debido a la homogenidad de la población

Antitumor and Antiviral Activity of Colombian Medicinal Plant Extracts

Extracts of nine species of plants traditionally used in Colombia for the treatment of a variety of diseases were tested in vitro for their potential antitumor (cytotoxicity) and antiherpetic activity. MTT (Tetrazolium blue) and Neutral Red colorimetric assays were used to evaluate the reduction of viability of cell cultures in presence and absence of the extracts. MTT was also used to evaluate the effects of the extracts on the lytic activity of herpes simplex virus type 2 (HSV-2). The 50 percent cytotoxic concentration (CC50) and the 50 percent inhibitory concentration of the viral effect (EC50) for each extract were calculated by linear regression analysis. Extracts from Annona muricata, A. cherimolia and Rollinia membranacea, known for their cytotoxicity were used as positive controls. Likewise, acyclovir and heparin were used as positive controls of antiherpetic activity. Methanolic extract from Annona sp. on HEp-2 cells presented a CC50 value at 72 hr of 49.6x103mg/ml. Neither of the other extracts examined showed a significant cytotoxicity. The aqueous extract from Beta vulgaris, the ethanol extract from Callisia grasilis and the methanol extract Annona sp. showed some antiherpetic activity with acceptable therapeutic indexes (the ratio of CC50 to EC50). These species are good candidates for further activity-monitored fractionation to identify active principles

The production of MLR-blocking factors after lymphocyte immunotherapy for RSA does not predict the outcome of pregnancy.

PROBLEM: The questions of whether production of mixed lymphocyte reaction-blocking factors (MLR-BFs) after immunotherapy with lymphocytes for recurrent spontaneous abortion (RSA) has prognostic value and whether cytotoxic antibodies are also involved were tested. METHOD OF STUDY: A prospective study with 33 patients who had a history of two or more abortions, lacking MLR-BFs, was carried out. The patients received immunizations with lymphocytes and 6 weeks or later were tested for seroconversion of MLR-BFs. Seventeen of these thirty-three patients were evaluated for antipaternal cytotoxic antibodies. The results were correlated with the outcome of the next pregnancy after treatment. RESULTS: Eighty percent of the 33 patients had a live child. Of those patients having success, only 50% produced MLR-BFs. Of those patients having a new loss, five did and two did not produce MLR-BF (P > 0.05). Regarding the 17 patients tested for cytotoxic antibodies, 4 of the 5 patients who tested positive had a new abortion, whereas only 1 of 12 whose tests remained negative did not have gestational success (P < 0.01). CONCLUSION: The presence of MLR-BFs is not a prognostic criterium for the outcome of pregnancy after alloimmunotherapy, and, consequently, it is not a good diagnostic tool for RSA of alloimmune cause.

Clinical features of early-onset Alzheimer disease in a large kindred with an E280A presenilin-1 mutation.

OBJECTIVES: To characterize clinical features of a very large pedigree with early-onset Alzheimer disease (AD) in which all affected individuals carry the identical glutamic acid-to-alanine mutation at codon 280 in the presenilin-1 gene. DESIGN: Clinical histories were obtained by patient and family interviews and through medical or civil records. Using standard diagnostic criteria, a case series of 128 individuals was identified, of which 6 have definitive (autopsy-proven) early-onset AD, 93 have probable early-onset AD, and 29 have possible early-onset AD. SETTING: Community based in Antioquia, Colombia. PATIENTS: A population-based sample in which all members of 5 extended families (nearly 3000 individuals) were surveyed. Criteria for inclusion required obtaining sufficient information to categorize the individual as affected. MAIN OUTCOME MEASURES: Age at onset, neuropsychological profile, neurologic history, and examination. RESULTS: The patients had a mean age at onset of 46.8 years (range, 34-62 years). The average interval until death was 8 years. Headache was noted in affected individuals significantly more frequently than in those not affected. The most frequent presentation was memory loss followed by behavior and personality changes and progressive loss of language ability. In the final stages, gait disturbances, seizures, and myoclonus were frequent. CONCLUSIONS: Other than the early onset, this clinical phenotype is indistinguishable from sporadic AD except that affected individuals frequently complained of headache preceding and during the disease. Despite the uniform genetic basis for the disease, there was significant variability in the age at onset, suggesting an important role for environmental factors or genetic modifiers in determining the age at onset.

Prion plaques: molecular tumors. A hypothesis on the etiopathogenesis of prion diseases.

In spite of remarkable advances in the etiopathogenesis of spongiform encephalopathies in man and animals and the growing acceptance of the prion hypothesis, there is no explanation for the supposed 'autocatalytic' activity of this protein molecule. Our molecular tumor hypothesis proposes that the prion protein is a genotoxin which interacts directly or indirectly but specifically with its homologous cellular gene introducing mutations which lead to aberrant processing and accumulation of the protein. It is also speculated that this hypothesis would shed some light on other diseases not presently classified as prion diseases and in the process of ageing.

Is the immune system necessary for placental reproduction? A hypothesis on the mechanisms of alloimmunotherapy in recurrent spontaneous abortion.

Recurrent spontaneous abortion is being treated with alloimmunotherapy which consists of the inoculation of allogeneic mononuclear cells. However the mechanisms explaining the benefits of this therapy are not clear yet. Taking the immunotrophic hypothesis as the paradigm of the field of reproductive immunology, hereby we present a hypothesis to propose a role for the immune system in reproduction and at the same time to explain how alloimmunotherapy may work. We base our view on several facts: first, immunodeficient mice reproduce--albeit their progeny may not be as robust and numerous as that from normal individuals. Second, maternal lymphocytes cross the placenta and may induce graft versus host disease. Third, graft versus host disease in newborn F1 mice inoculated with paternal lymphocytes can be prevented by inoculation of the mother with the same paternal lymphocytes before and during pregnancy. We propose that: 1) the immune system by itself is not necessary for placental reproduction; 2) the immune system plays its major role in reproduction by counteracting the allogeneic response generated against the fetus; 3) recurrent spontaneous abortion represents a type of graft versus host disease induced by maternal cells infiltrating the fetus; and 4) alloimmunotherapy induces an antiidiotypic response necessary to counteract the graft versus host reaction in the fetus.

Role of splenectomy in prevention of hemorrhagic enteritis and death from hemorrhagic enteritis virus in turkeys.

Hemorrhagic diarrhea, gross hemorrhagic enteritis, and death caused by intravenous virus injection of hemorrhagic enteritis virus (HEV) were prevented in otherwise susceptible turkey poults by surgical splenectomy. The splenectomized poults produced anti-HEV antibodies, which indicated that splenectomy did not completely prevent replication of the virus. These results indicate that the spleen is necessary for the development of the intestinal lesions of this disease. The role of a toxic factor in this disease is discussed.

Hemorrhagic enteritis in turkeys: purification and quantification of the virus.

Two methods for purifying the virus of hemorrhagic enteritis from infected turkey spleens are described. One procedure utilized precipitation with polyethylene glycol, and the other consisted of trichlorotrifluoroethane extraction. Both procedures included sucrose-cesium chloride gradient centrifugation in the final purification step. The buoyant density of the viral fraction was 1.34 g/cm3, typical for adenoviral particles, and the size and morphologic characteristics of the virions observed by transmission electron microscopy suggested that the purified virus belongs to the family Adenoviridae. The biologic activity of the purified virus was titrated by inoculating 10-fold dilutions of the viral suspension into turkey poults. Mortality and hemorrhagic diarrhea proved to be inconsistent parameters of infection, and the degree of splenomegaly was proportional to the virus dose. The body/spleen ratio was the parameter selected for measuring viral activity, and the body/spleen ratio 50% was adopted as the unit for the titration of the virus. By using the same system it was demonstrated that the infectivity of the virus could be neutralized with antiserum produced in turkeys.