Identification of a tumor-specific allo-HLA-restricted γδTCR.

γδT cells are key players in cancer immune surveillance because of their ability to recognize malignant transformed cells, which makes them promising therapeutic tools in the treatment of cancer. However, the biological mechanisms of how γδT-cell receptors (TCRs) interact with their ligands are poorly understood. Within this context, we describe the novel allo-HLA-restricted and CD8α-dependent Vγ5Vδ1TCR. In contrast to the previous assumption of the general allo-HLA reactivity of a minor fraction of γδTCRs, we show that classic anti-HLA-directed, γδTCR-mediated reactivity can selectively act on hematological and solid tumor cells, while not harming healthy tissues in vitro and in vivo. We identified the molecular interface with proximity to the peptide-binding groove of HLA-A*24:02 as the essential determinant for recognition and describe the critical role of CD8 as a coreceptor. We conclude that alloreactive γδT-cell repertoires provide therapeutic opportunities, either within the context of haplotransplantation or as individual γδTCRs for genetic engineering of tumor-reactive T cells.

Optimal nutrition and the ever-changing dietary landscape: a conference report.

The field of nutrition has evolved rapidly over the past century. Nutrition scientists and policy makers in the developed world have shifted the focus of their efforts from dealing with diseases of overt nutrient deficiency to a new paradigm aimed at coping with conditions of excess-calories, sedentary lifestyles and stress. Advances in nutrition science, technology and manufacturing have largely eradicated nutrient deficiency diseases, while simultaneously facing the growing challenges of obesity, non-communicable diseases and aging. Nutrition research has gone through a necessary evolution, starting with a reductionist approach, driven by an ambition to understand the mechanisms responsible for the effects of individual nutrients at the cellular and molecular levels. This approach has appropriately expanded in recent years to become more holistic with the aim of understanding the role of nutrition in the broader context of dietary patterns. Ultimately, this approach will culminate in a full understanding of the dietary landscape-a web of interactions between nutritional, dietary, social, behavioral and environmental factors-and how it impacts health maintenance and promotion.

The relationship between liver stiffness measurement and outcome in patients with chronic hepatitis C and cirrhosis: a retrospective longitudinal hospital study.

BACKGROUND: There is a relationship between liver stiffness measurement (LSM) and outcome of HCV patients. AIM: To evaluate the performance of LSM to predict outcome of HCV patients at risk of liver-related complication. METHODS: We established a retrospective longitudinal cohort of 341 HCV patients with unequivocal cirrhosis. All underwent LSM and were followed from September 2006 to July 2015. Outcome measure was a composite end-point of end-stage liver disease (ESLD) and/or hepatocellular carcinoma (HCC). Cox models and areas under receiver operating characteristic (AUROC) curves were used to evaluate independent risk factors of outcome. RESULTS: Overall, LSM was below the 12.5 kPa threshold in 129 (37.8%) patients, including three-fourth and one-third of patients with or without a sustained virological response respectively. Liver disease progressed in 136 (39.9%) patients after a median observational period of 23.5 months. Older age, male gender, alcohol use disorders, metabolic syndrome and LSM were independent risk factors of liver disease progression. Age, alcohol use disorders and LSM were independently associated with ESLD. Age, gender and metabolic syndrome, but not LSM, were associated with HCC. The AUROC curves for disease progression, ESLD and HCC were 0.67, 0.70 and 0.58 respectively. Patients with a liver stiffness >12.5 kPa were at the highest risk of liver disease progression; below 12.5 kPa, liver stiffness was not discriminant. CONCLUSION: Liver stiffness measurement is not a surrogate of disease progression of HCV patients with cirrhosis. HCV patients with cirrhosis should undergo the recommended follow-up, regardless of liver stiffness measurement.

Loss of DJ-1 impairs antioxidant response by altered glutamine and serine metabolism.

The oncogene DJ-1 has been originally identified as a suppressor of PTEN. Further on, loss-of-function mutations have been described as a causative factor in Parkinson's disease (PD). DJ-1 has an important function in cellular antioxidant responses, but its role in central metabolism of neurons is still elusive. We applied stable isotope assisted metabolic profiling to investigate the effect of a functional loss of DJ-1 and show that DJ-1 deficient neuronal cells exhibit decreased glutamine influx and reduced serine biosynthesis. By providing precursors for GSH synthesis, these two metabolic pathways are important contributors to cellular antioxidant response. Down-regulation of these pathways, as a result of loss of DJ-1 leads to an impaired antioxidant response. Furthermore, DJ-1 deficient mouse microglia showed a weak but constitutive pro-inflammatory activation. The combined effects of altered central metabolism and constitutive activation of glia cells raise the susceptibility of dopaminergic neurons towards degeneration in patients harboring mutated DJ-1. Our work reveals metabolic alterations leading to increased cellular instability and identifies potential new intervention points that can further be studied in the light of novel translational medicine approaches.

Non-traumatic subdural hematoma secondary to septic brain embolism: A rare cause of unexpected death in a drug addict suffering from undiagnosed bacterial endocarditis.

Acute subdural hematomas are mostly due to blunt traumatization of the head. In rare instances, subdural bleeding occurs without evidence of a previous trauma following spontaneous hemorrhage, e.g. from a ruptured aneurysm or an intracerebral hematoma perforating the brain surface and the arachnoid. The paper presents the morphological, microbiological and toxicological findings in a 38-year-old drug addict who was found by his partner in a dazed state. When brought to a hospital, he underwent trepanation to empty a right-sided subdural hematoma, but he died already 4h after admission. Autopsy revealed previously undiagnosed infective endocarditis of the aortic valve as well as multiple infarctions of brain, spleen and kidneys obviously caused by septic emboli. The subdural hematoma originated from a subcortical brain hemorrhage which had perforated into the subdural space. Microbiological investigation of the polypous vegetations adhering to the aortic valve revealed colonization by Streptococcus mitis and Klebsiella oxytoca. According to the toxicological analysis, no psychotropic substances had contributed to the lethal outcome. The case reported underlines that all deaths of drug addicts should be subjected to complete forensic autopsy, as apart from intoxications also natural and traumatic causes of death have to be taken into consideration.

Immuno-histochemical study of ovine cystic echinococcosis (Echinococcus granulosus) shows predominant T cell infiltration in established cysts.

Ovine hydatidosis (OH; Echinococcus granulosus) is endemic in several European countries surrounding the Mediterranean basin. There have been a limited number of studies aimed at evaluating the local immune response to established tissue cysts in the ovine host. In the present study, immunohistochemical analysis of lymphocyte populations surrounding established cysts showed a predominance of CD3+ T cells compared to CD79+ B cells. A percentage of infiltrating lymphocytes were also FoxP3+, suggesting that established ovine cysts may be protected from immune aggression through the suppressive action of T regulatory cells. The present study contributes to the understanding of local immune responses to ovine echinococcosis.

Effect of maternal tobacco smoking or exposure to second-hand smoke on the levels of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine of mother and the first urine of newborn.

Tobacco smoking during pregnancy is associated with a variety of negative consequences not only for the mother, but also for the developing fetus. Many studies have shown that carcinogens contained in tobacco smoke permeate across the placenta, and are found in fetus. The aim of the study was to determine the prenatal exposure to tobacco-specific carcinogenic N-nitrosamines on the basis of measurements of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine of smoking and second-hand smoke (SHS) exposed women and in the first urine of their newborns. A questionnaire documenting demographics and socio-economical data, smoking habits and exposure to SHS was completed by 121 delivering women near or at term. Maternal concentrations of cotinine and NNAL were measured in urine of the mother and the first urine of her newborn infant by liquid chromatography tandem mass spectrometry (LC/MS/MS). The mean concentration of cotinine was 439.2 ng/mg creatinine and NNAL concentration in urine of smoking women was 74.0 pg/mg creatinine, and for her newborn 78.6 pg/mg creatinine. Among mothers exposed to SHS, cotinine and NNAL mean concentration were 23.1 ng/mg creatinine, and 26.4 pg/mg creatinine. In newborns of SHS exposed mothers during pregnancy the mean concentration of NNAL was 34.1 pg/mg creatinine, respectively. Active tobacco smoking as well as passive exposure to smoking during pregnancy is an important source of tobacco specific N-nitrosamines to the fetuses as evidenced by increased concentrations of this carcinogen. Determination of NNAL in maternal urine samples can be a useful biomarker of prenatal exposure of newborn to carcinogenic nitrosamines.

[Curriculum and expert courses on pacemaker and ICD therapy]. / Curriculum und Sachkundekurse Herzschrittmacher- und ICD-Therapie.

The curricula "Practice of Pacemaker Therapy" and "Practice of ICD Therapy" have been developed from practical experience with the first educational courses which are necessary to fulfill the German requirements of the medical products law which restricts the application of medical products to persons with the necessary education, knowledge and experience. The corresponding courses of competence under the auspices of the German Cardiac Society derive from this legal prerequisite. Competence refers to technical knowledge in cardiac implantable electrical devices (CIEDs) and understanding of possible dysfunctions as well as substantial knowledge on arrhythmia. The two curricula form the theoretical basis for the application of CIEDs. These courses represent an offer to cardiologists and all other physicians who wish to acquire and document competency in this field. A legal obligation to participate in these competency courses does not currently exist in Germany as long as evidence can be provided that this competency has been achieved by other means. Both curricula have proven to be comprehensive and practically highly useful and have been presented by highly committed specialists with expertise in this topic at a high level. Since 2005 some 2,000 physicians have been trained in courses on pacemaker therapy and more than 1,000 physicians in courses on ICD therapy with an ongoing high level demand to be expected in the future.

Disseminated histoplasmosis in a cat in Europe.

A cat was presented with a history of vomiting, decreased appetite and weight loss. Abnormal findings were poor body condition, pale mucous membranes, dehydration and a palpable abdominal mass. Abdominal ultrasound showed lymph node enlargement, a mass of uncertain origin, thickening of the muscularis layer of the small bowel, focal thickening of the ileum with loss of layering and free peritoneal fluid. Cytology revealed a piogranulomatous infiltrate and numerous macrophages containing oval or round yeast-like cells 2 to 5 microm diameter with a central, spherical, lightly basophilic body surrounded by a clear halo, compatible with Histoplasma capsulatum, within the cytoplasm. Post-mortem examination revealed cavity effusions, granulomatous nodules in lungs, intestine and omentum, thickened intestinal walls and intestinal perforation. Staining with Grocott and immunohistochemistry (IHC) revealed numerous organisms within the granulomatous reaction. H. capsulatum has a worldwide distribution in temperate and subtropical climates. To the author's knowledge, this is the first report of feline histoplasmosis in Europe.

Canine bladderworm (Capillaria plica) infection associated with glomerular amyloidosis.

Capillaria plica (Trichuroidea: Capillariidae), commonly known as bladderworm, is a nematode rarely associated with clinical disease that resides in the lower urinary tract of wild and domestic canids. In the present paper a case of canine urinary capillariosis associated with glomerular amyloidosis is described. The dog, an 8-year-old, male, hunting Jagd terrier had a history of weight loss and diarrhoea and was referred to the University of Parma Teaching Veterinary Hospital (UPTVH). Clinical and laboratory tests shown here suggest that C. plica may be a contributing factor to glomerular amyloidosis.

MEK inhibitors potentiate dexamethasone lethality in acute lymphoblastic leukemia cells through the pro-apoptotic molecule BIM.

Glucocorticoids (GCs) are common components of many chemotherapeutic regimens for lymphoid malignancies. GC-induced apoptosis involves an intrinsic mitochondria-dependent pathway. We and others have shown that BIM (BCL-2 interacting mediator of cell death), a BH3-only pro-apoptotic protein, is up-regulated by dexamethasone (Dex) treatment in acute lymphoblastic leukemia (ALL) cells and plays an essential role in Dex-induced apoptosis. Furthermore, BIM is inactivated by extracellular signal-regulated kinase (ERK)-mediated phosphorylation. We therefore hypothesized co-treatment with Dex and MEK/ERK inhibitors would promote apoptosis in ALL cells through BIM up-regulation and activation. We show here that MEK inhibitors (PD184352 and PD98059) synergistically enhance Dex lethality in a variety of ALL cells and in two primary ALL specimens. Co-treatment with Dex and PD184352 results in BIM accumulation, pro-apoptotic BAX/BAK activation, and cytochrome c release from mitochondria. Down-regulation of BIM by short-hairpin RNA (shRNA) in ALL cells suppressed BAX/BAK activation, cytochrome c release, and cell death by Dex/PD184352 co-treatment. BIM accumulated by this treatment sequesters anti-apoptotic BCL-XLMCL-1, resulting in the release of BAK from these anti-apoptotic molecules. This study provides a rational foundation for future attempts to improve the activity of GCs with clinically relevant pharmacologic MEK inhibitors in the treatment of ALL and possibly other hematologic malignancies.

[Training requirements for transvenous implantation of pacemakers and cardioverter-defibrillators]. / Empfehlungen zur strukturierung der herzschrittmacher- und defibrillatortherapie.

Therapy with implantable pacemakers, cardioverter defibrillators (ICD), and devices for cardiac resynchronization (CRT) is performed by various medical and surgical specialists. With the change from implantation by thoracotomy to the transvenous approach, an increasing number of devices are implanted by cardiologists. The purpose of this paper is to establish training requirements for transvenous device therapy, implantation and follow-up examinations, regardless of the implanting person, an internist, cardiologist, general surgeon, or cardiothoracic surgeon. Epicardial lead placement should be performed only by surgeons. Two levels of training topics are defined, level 1 for pacemakers and level 2 for ICD and CRT devices. Surgery that involves the implantation of foreign material should demand the highest standards of operating rooms design and environment. Catheter laboratories used for implantations should meet operating room standards. Complications need to be documented carefully for quality control.

Past obstetric history and risk of malignant breast neoplasms.

Many studies indicate hormonal disorders as a crucial reason for breast pathology. They are also probably responsible for the development of benign neoplasms and play a role in the origin and development of breast carcinoma. Although the mammary gland is under the influence of many steroid and peptide hormones such as thyroid hormones, prolactin, growth hormone, glucocorticosteroids, it is estrogen that plays an important role in the development of breast cancer. The purpose of the study was to analyze the obstetrical past of patients and the potential influence on the risk of developing malignant breast neoplasms. The participants in the study were healthy women with no changes in mammary glands (control group) and women with diagnosed malignant or benign breast neoplasms (study group). The total number of participants was 555 females aged 35-70 years. The study was carried out in the Greater Poland and Lubuskie province between 2005 and 2006. Hormonal disorders in childhood and puberty symptoms of early menarche play a crucial role in increasing the risk of malignant breast neoplasms. In women who experienced one or more miscarriages the risk of malignant breast neoplasms is significantly increased. On the basis of our study we calculated the odds ratio (OR) of malignant breast neoplasms among women who during lactation experienced problems needing medical intervention (OR = 2.25; 95% CI, 1.20-4.19) in comparison to women who experienced no problems).

Wolbachia in Dirofilaria repens, an agent causing human subcutaneous dirofilariasis.

Human subcutaneous dirofilariasis is an increasingly reported zoonosis caused by several filarial species, in particular by Dirofilaria (Nochtiella) repens. Like many filarial worms, D. repens harbors the bacterial endosymbiont Wolbachia that has been implicated in the inflammatory features of filarial infection. Immunohistochemical staining against the Wolbachia surface protein (WSP) was carried out on 14 skin nodules and showed numerous bacteria within the intact worms and occasional positive staining within the surrounding inflammatory infiltrate. Serum samples from 11 of these patients resulted positive for total immunoglobulin G titers against WSP as examined in enzyme-linked immunosorbent assay. This is the first description of Wolbachia distribution in D. repens and the first report of specific immune response to Wolbachia in patients with subcutaneous dirofilariasis.

[Evaluation of continuing medical education (CME) in print media]. / Evaluation der Fortbildungin Printmedien.

BACKGROUND AND OBJECTIVE: In order to achieve points for the CME certificate of the German Medical Council ("Arztekammer") the reader of a CME article in a medical journal has to fill in an evaluation form that includes knowledge assessment. This article summarizes the data of the first ten CME presentations in the "Deutsche Medizinische Wochenschrift". METHODS: 4481 completed data sets were evaluated by chi-square and binomial tests using SPSS version 10.0. RESULTS: 85% of the participants were specialists, mainly in internal medicine or surgery: those working in private practice or a hospital took part. The great majority had received their licence one to 30 years ago. The topics dealt with diseases which were not uniformly often seen in clinical practice. The individually perceived changes in the strategy of diagnosis and therapy, induced by the CME article, was mainly dependent on qualification and specialization of the reader as well as on the frequency, with which the reader had been treating the respective disorder. The articles were appreciated by specialists as well as by non-specialists. Knowledge assessment was largely made on the basis of the article alone and was successfully passed by nearly 90% of the participants. CONCLUSION: For the first time this article provides detailed data on CME activities in a nation-wide available German medical journal and thus forms the basis for discussing further the definition of quality criteria for CME articles in medical journals.

Partial genetic characterization of West Nile virus strains, New York State, 2000.

We analyzed nucleotide sequences from the envelope gene of 11 West Nile (WN) virus strains collected in New York State during the 2000 transmission season to determine whether they differed genetically from each other and from the initial strain isolated in 1999. The complete envelope genes of these strains were amplified by reverse transcription-polymerase chain reaction. The resulting sequences were aligned, the genetic distances were computed, and a phylogenetic tree was constructed. Ten (0.7%) of 1,503 positions in the envelope gene were polymorphic in one or more sequences. The genetic distances were 0.003 or less. WN virus strains circulating in 2000 were homogeneous with respect to one another and to a strain isolated in 1999.

Liver penetration by a duodenal ulcer in a young woman.

Liver penetration is a rare but serious complication of peptic ulcer disease. We report a case of a 33-year-old woman who took large doses of nonsteroidal antiinflammatory drugs and developed a giant duodenal ulcer that penetrated into her liver. The diagnosis was based on histologic examination of endoscopic biopsies. She was initially treated with a proton pump inhibitor, but, within 5 weeks, she developed a symptomatic postbulbar stricture that required surgical correction. We also review 11 other reported cases of endoscopically and histologically diagnosed peptic ulcer penetration into the liver.

Primary cardiac rhabdomyosarcoma in a cat.

A seven-year-old domestic shorthair (DSH) cat was presented with anorexia and dyspnea. Pleural-pericardial effusion was detected with thoracic radiographs and echocardiography. Echocardiography demonstrated a large, soft-tissue mass in the right ventricular wall, protruding both into the pericardial space and into the right ventricle. Postmortem examination findings included a large mass in the right ventricular wall and multiple smaller masses on the external surface of the left ventricle and on the internal surface of the pericardium. Results of the histopathological and immunohistochemical examinations of the masses were consistent with rhabdomyosarcoma. This is the first reported case of primary cardiac rhabdomyosarcoma in the cat.

Interactions between 2-fluoroadenine 9-beta-D-arabinofuranoside and the kinase inhibitor UCN-01 in human leukemia and lymphoma cells.

Interactions between the purine analogue 2-fluoroadenine 9-beta-D-arabinofuranoside (F-ara-A) and the kinase inhibitor UCN-01 have been examined in human leukemia cells (U937 and HL-60) with respect to induction of mitochondrial damage, caspase activation, apoptosis, and loss of clonogenic survival. Simultaneous or subsequent exposure of F-ara-A-treated cells (2 microM) to UCN-01 (100 nM) resulted in a marked potentiation of apoptosis, manifested by loss of mitochondrial membrane potential (delta psi(m)), cleavage/activation of procaspase-9 and procaspase-3, DNA fragmentation, and degradation of poly-ADP(ribosyl) polymerase. Coadministration of UCN-01 with F-ara-A was also associated with diminished phosphorylation of the cdc25 phosphatase. In contrast, exposure of cells to the sequence UCN-01, followed by F-ara-A, resulted in only a modest increase in apoptotic cells. The ability of UCN-01 to potentiate F-ara-A-mediated lethality was not mimicked by the selective PKC inhibitor bisindolylmaleimide, nor did treatment of cells with UCN-01 enhance formation of F-ara-ATP or increase incorporation of [3H]F-ara-A into DNA. Enhanced apoptosis in cells exposed sequentially or simultaneously to F-ara-A and UCN-01 was accompanied by a substantial reduction in colony formation (e.g., to 0.01% of control values). Cotreatment with UCN-01 also increased F-ara-A-mediated apoptosis and loss of delta psi(m) in U937 cells ectopically expressing Bcl-2, although not to the same extent as that observed in empty-vector controls. Finally, simultaneous exposure (24 h) of malignant B lymphocytes from the pleural effusion of a patient with indolent non-Hodgkin's lymphoma to F-ara-A and UCN-01 ex vivo resulted in a striking increase in apoptosis, as determined by terminal deoxynucleotidyltransferase-mediated nick end labeling assay. These findings indicate that UCN-01 increases F-ara-A-induced mitochondrial damage and apoptosis in human leukemia cells in a sequence-dependent manner, and that these events occur in at least some primary human lymphoma cells.