Induction of apoptosis by adenovirus E4orf4 protein is specific to transformed cells and requires an interaction with protein phosphatase 2A

We previously have shown that adenovirus type 5 E4orf4 protein associates with protein phosphatase 2A (PP2A) and induces apoptosis in transformed cells in a p53-independent manner. Here we show that the interaction between E4orf4 and PP2A is required for induction of apoptosis by the viral protein. This conclusion is supported by a mutation analysis of E4orf4 protein, showing a correlation between the ability to bind PP2A and to induce apoptosis, and by the observation that transfection of an antisense construct of the PP2A-B55 subunit reduces expression of the PP2A-B55 subunit and inhibits induction of apoptosis by E4orf4, but not by p53. The mutant analysis also indicates that even a low level of interaction with PP2A is sufficient to initiate the E4orf4 apoptotic pathway. In addition, E4orf4 inhibits cellular transformation by various oncogenes, and this function is coupled to its ability to induce apoptosis. Furthermore, expression of oncogenes in primary cell cultures sensitizes these cells to induction of apoptosis by E4orf4. Our results suggest that E4orf4 is a potentially useful tool for cancer gene therapy.     

Intermodal matching of emotional expressions in young children with autism

This study examined the ability of young children with autism spectrum disorders (ASD) to detect affective correspondences between facial and vocal expressions of emotion using an intermodal matching paradigm. Four-year-old children with ASD (n = 18) and their age-matched normally developing peers (n = 18) were presented pairs of videotaped facial expressions accompanied by a single soundtrack matching the affect of one of the two facial expressions. In one block of trials, the emotions were portrayed by their mothers; in another block of trials, the same emotion pairs were portrayed by an unfamiliar woman. Findings showed that ASD children were able to detect the affective correspondence between facial and vocal expressions of emotion portrayed by their mothers, but not a stranger. Furthermore, in a control condition using inanimate objects and their sounds, ASD children also showed a preference for sound-matched displays. These results suggest that children with ASD do not have a general inability to detect intermodal correspondences between visual and vocal events, however, their ability to detect affective correspondences between facial and vocal expressions of emotions may be limited to familiar displays.     

AN EXPERIMENTAL STUDY OF PAROTITIS (MUMPS)

Cats injected into the parotid gland and testicle with a bacterial sterile filtrate of the salivary secretion of children in the active stage of parotitis or mumps can be made to develop a pathological condition having several points of resemblance to the condition present in mumps in human beings. After an incubation stage of from five to eight days definite changes have been noted in the temperature, blood leukocytes, and inoculated organs. The temperature rise begins within twenty-four hours of the inoculations and reaches a maximum in from seven to fourteen days. The febrile rise fluctuates between 1° and 2.5° C. The white blood cells begin to increase on the second day following the inoculation. The first change is a polymorphonuclear leukocytosis which precedes the glandular swellings. This initial rise is followed by a decline, after which the lymphocytes increase. The increase is confined to the small lymphocytes, which increase to from 7 to 10 per cent of their initial number. The inoculated glands become swollen and tender. The swelling and tenderness become apparent from the fifth to the ninth days and persist for a variable period. The parotid changes are less constant or less obvious than are the testicular. The latter are constant and endure from eight to twelve days. The rise of temperature and the leukocytosis precede the glandular swelling, but all the changes reach the maximum at about the same time, after which they decline gradually. What may be regarded as normal conditions are reestablished in four weeks or less. The intraparotid and intratesticular injections of extracts of normal parotid gland and testicles may cause a mild rise of temperature and leukocytosis of brief duration, but swelling and tenderness are absent. The white cells increased are the polymorphonuclears and not the lymphocytes. The intraparotid and intratesticular injections of filtrates of normal saliva may cause a mild rise of temperature of very brief duration, but leukocytosis, swelling, and tenderness do not appear. The histological changes in the parotid gland when present consist chiefly of edema of the interlobular connective tissue with mononuclear interstitial infiltration about the ducts and elsewhere. In cases of long duration the ducts may be dilated. But in some instances the swollen gland while showing congestion and edema in gross showed inconspicuous changes under the microscope. The histological changes in the testicle are of two kinds: inconstant changes of cellular invasion between the tubules and swelling or even multiplication of the interstitial cells, constant ones consisting of degeneration of the epithelium and interference with spermatogenesis, a condition to which we have applied the term "spermatorrhexis." The pathological conditions set up by the filtrate derived from the salivary secretion of cases of acute parotitis are intensified by successive transfers through a small series of cats of the extract and emulsion of the parotid gland and testicle previously inoculated. The pathological changes are also prevented or reduced when the extract or emulsion is previously incubated with a quantity of blood serum obtained from a cat which has survived inoculation. Normal serum, on the other hand, has no such inhibiting effect. The deduction from these experiments is to the effect that the salivary secretion in parotitis or mumps contains a filterable substance capable of setting up a series of definite pathological conditions when inoculated into the testicle and parotid glands of cats. Whether this active material is a microorganism and if so whether it is the specific microbic cause of parotitis or mumps remains to be ascertained.     

Understanding hyperlipidemia and atherosclerosis: lessons from genetically modified apoe and ldlr mice.

Hyperlipidemia is the most important risk factor for atherosclerosis, which is the major cause of cardiovascular disease. The etiology of hyperlipidemia and atherosclerosis is complex and governed by multiple interacting genes. However, mutations in two genes have been shown to be directly involved, i.e., the low-density lipoprotein receptor (LDLR) and apolipoprotein E (ApoE). Genetically modified mouse models have been instrumental in elucidating the underlying molecular mechanisms in lipid metabolism. In this review, we focus on the use of two of the most widely used mouse models, ApoE- and LDLR-deficient mice. After almost a decade of applications, it is clear that each model has unique strengths and drawbacks when carrying out studies of the role of additional genes and environmental factors such as nutrition and lipid-lowering drugs. Importantly, we elaborate on mice expressing mutant forms of APOE, including the APOE3Leiden ( APOE3L ) and the APOE2 knock-in ( APOE 2k) mouse models. These models have outstanding potential, as they are highly responsive to dietary factors and pharmacological interventions.     

Kaposi's sarcoma-associated herpesvirus: clinical, diagnostic, and epidemiological aspects

Kaposi's sarcoma-associated herpesvirus (KSHI) is one of the few viruses proven to be associated with tumorigenesis in humans. Its causal association with all clinical and epidemiological variants of Kaposi's sarcoma (KS) is well established. KSHV is also involved in the pathogenesis of primary effusion lymphoma (PEL) and a subset of multicentric Castleman's disease (MCD). Possible associations of KSHV with other clinical settings have been extensively examined. The findings from several of these studies are contradictory and are yet to be resolved. Concentrated effort over the last decade, since the initial discovery of KSHV, led to the development of several experimental systems that resulted in a better comprehension of the biological characteristics of KSHV and set the stage for the understanding of mechainisms by which diseases are induced by the virus. The development of molecular, histological, and serological tools for KSHV diagnosis allowed researchers to track the transmission and to study the epidemiology of KSHV. These assays have been applied, in particular in ambiguous cases, in order to confirm clinically and pathologically based diagnoses. Here, we review the advances in the clinical, experimental, diagnostic, and epidemiological research of KSHV.     

Establishing a high level of knowledge regarding bioterrorist threats in emergency department physicians: methodology and the results of a national bio-preparedness project

INTRODUCTION: Medical systems worldwide are facing the new threat of morbidity associated with the deliberate dispersal of microbiological agents by terrorists. Rapid diagnosis and containment of this type of unannounced attack is based on the knowledge and capabilities of medical staff. In 2004, the knowledge of emergency department physicians of anthrax was tested. The average test score was 58%. Consequently, a national project on bioterrorism preparedness was developed. The aim of this article is to present the project in which medical knowledge was enhanced regarding a variety of bioterrorist threats, including cutaneous and pulmonary anthrax, botulinum, and smallpox. METHODS: In 2005, military physicians and experts on bioterrorism conducted special seminars and lectures for the staff of the hospital emergency department and internal medicine wards. Later, emergency department senior physicians were drilled using one of the scenarios. RESULTS: Twenty-nine lectures and 29 drills were performed in 2005. The average drill score was 81.7%.The average score of physicians who attended the lecture was 86%, while those who did not attend the lectures averaged 78.3% (NS). CONCLUSIONS: Emergency department physicians were found to be highly knowledgeable in nearly all medical and logistical aspects of the response to different bioterrorist threats. Intensive and versatile preparedness modalities, such as lectures, drills, and posters, given to a carefully selected group of clinicians, can increase their knowledge, and hopefully improve their response to a bioterrorist attack.     

RBL cells as models for in vitro studies of mast cells and basophils

Since their establishment in 1981, RBL-2H3 cells have been widely used as a mast cell (MC) model. Their ability to be easily grown in culture in large amounts, their responsiveness to FcεRI-mediated triggers and the fact that they can be genetically manipulated, have provided advantages over primary MCs, in particular for molecular studies relying on genetic screening. Furthermore, the ability to generate clones that stably express proteins of interest, for example, a human receptor, have marked the RBL cells as an attractive MC model for drug screening. Indeed, 3 RBL reporter cell lines (RS-ATL8, NFAT-DsRed, and NPY-mRFP) have been generated providing useful models for drug and allergen screening. Similarly, RBL cells stably expressing the human MrgprX2 receptor provide a unique paradigm for analyzing ligand interactions and signaling pathways of the unique human receptor. Finally, transient co-transfections of RBL cells allow functional genomic analyses of MC secretion by combining library screening with simultaneous expression of a reporter for exocytosis. RBL cells thus comprise powerful tools for the study of intracellular membrane trafficking and exocytosis and the detection of allergens, vaccine safety studies and diagnosis of allergic sensitization. Their recent uses as an investigative tool are reviewed here.     

Combined Haploidentical and Umbilical Cord Blood Allogeneic Stem Cell Transplantation for High-Risk Lymphoma and Chronic Lymphoblastic Leukemia

Limited studies have reported on outcomes for lymphoid malignancy patients receiving alternative donor allogeneic stem cell transplants. We have previously described combining CD34-selected haploidentical grafts with umbilical cord blood (haplo-cord) to accelerate neutrophil and platelet engraftment. Here, we examine the outcome of patients with lymphoid malignancies undergoing haplo-cord transplantation at the University of Chicago and Weill Cornell Medical College. We analyzed 42 lymphoma and chronic lymphoblastic leukemia (CLL) patients who underwent haplo-cord allogeneic stem cell transplantation. Patients underwent transplant for Hodgkin lymphoma (n?=?9, 21%), CLL (n?=?5, 12%) and non-Hodgkin lymphomas (n?=?28, 67%), including 13 T cell lymphomas. Twenty-four patients (52%) had 3 or more lines of therapies. Six (14%) and 1 (2%) patients had prior autologous and allogeneic stem cell transplant, respectively. At the time of transplant 12 patients (29%) were in complete remission, 18 had chemotherapy-sensitive disease, and 12 patients had chemotherapy-resistant disease. Seven (17%), 11 (26%), and 24 (57%) patients had low, intermediate, and high disease risk index before transplant. Comorbidity index was evenly distributed among 3 groups, with 13 (31%), 14 (33%), and 15 (36%) patients scoring 0, 1 to 2, and ≥3. Median age for the cohort was 49 years (range, 23 to 71). All patients received fludarabine/melphalan/antithymocyte globulin conditioning regimen and post-transplant graft-versus-host disease (GVHD) prophylaxis with tacrolimus and mycophenolate mofetil. The median time to neutrophil engraftment was 11 days (range, 9 to 60) and to platelet engraftment 19.5 days (range, 11 to 88). Cumulative incidence of nonrelapse mortality was 11.6% at 100 days and 19 % at one year. Cumulative incidence of relapse was 9.3% at 100 days and 19% at one year. With a median follow-up of survivors of 42 months, the 3-year rates of GVHD relapse free survival, progression-free survival, and overall survival were 53%, 62%, and 65%, respectively, for these patients. Only 8% of the survivors had chronic GVHD. In conclusion, haplo-cord transplantation offers a transplant alternative for patients with recurrent or refractory lymphoid malignancies who lack matching donors. Both neutrophil and platelet count recovery is rapid, nonrelapse mortality is limited, excellent disease control can be achieved, and the incidence of chronic GVHD is limited. Thus, haplo-cord achieves high rates of engraftment and encouraging results.     

Clinical and Epidemiological Significance of Carbapenem Resistance in Acinetobacter baumannii Infections

Carbapenems are considered the treatment of choice for Acinetobacter baumannii infections. Many facilities implement preventive measures toward only carbapenem-resistant A. baumannii (CRAB). However, the independent role of the carbapenem resistance determinant on patient outcomes remains controversial. In a 6-year analysis of adults with A. baumannii bloodstream infection (BSI), the outcomes of 149 CRAB isolates were compared to those of 91 patients with carbapenem-susceptible A. baumannii. In bivariable analyses, CRAB BSIs were significantly associated with worse outcomes and with a delay in the initiation of appropriate antimicrobial therapy (DAAT). However, in multivariable analyses, carbapenem resistance status was no longer associated with poor outcomes, while DAAT remained an independent predictor. The epidemiological significance of A. baumannii should not be determined by its resistance to carbapenems.