Human primary auditory cortex: cytoarchitectonic subdivisions and mapping into a spatial reference system

The transverse temporal gyrus of Heschl contains the human auditory cortex. Several schematic maps of the cytoarchitectonic correlate of this functional entity are available, but they present partly conflicting data (number and position of borders of the primary auditory areas) and they do not enable reliable comparisons with functional imaging data in a common spatial reference system. In order to provide a 3-D data set of the precise position and extent of the human primary auditory cortex, its putative subdivisions, and its topographical intersubject variability, we performed a quantitative cytoarchitectonic analysis of 10 brains using a recently established technique for observer-independent definition of areal borders. Three areas, Te1.1, Te1.0, and Te1.2, with a well-developed layer IV, which represent the primary auditory cortex (Brodmann area 41), can be identified along the mediolateral axis of the Heschl gyrus. The cell density was significantly higher in Te1.1 compared to Te1.2 in the left but not in the right hemisphere. The cytoarchitectonically defined areal borders of the primary auditory cortex do not consistently match macroanatomic landmarks like gyral and sulcal borders. The three primary auditory areas of each postmortem brain were mapped to a spatial reference system which is based on a brain registered by in vivo magnetic resonance imaging. The integration of a sample of postmortem brains in a spatial reference system allows one to estimate the spatial variability of each cytoarchitectonically defined region with respect to this reference system. In future, the transfer of in vivo structural and functional data into the same spatial reference system will enable accurate comparisons of cytoarchitectonic maps of the primary auditory cortex with activation centers as established with functional imaging procedures.     

Screening for Blunt Cerebrovascular Injuries: The Essential Role of Computed Tomography Angiography

The implementation of aggressive diagnostics refuted the thesis that blunt cerebrovascular injuries (BCVI) are rare events. Given the estimates from recent studies, the prevalence may be as high as 1 per 100 among blunt multiple trauma patients. The morbidity and mortality of unrecognized and untreated BCVI is exceptionally high and warrants distinct efforts to detect these injuries during the primary trauma survey. The primary goal is to detect BCVI before neurological symptoms occur, and to introduce anticoagulation or antiplatelet therapy as appropriate. Index injuries such as cervical spine fractures increase the prior probability of disease, but are not helpful in ruling BCVI out. Computed tomography angiography (CTA) may represent the screening tool of choice, although there is still limited evidence about its accuracy. Pooled data from six studies (1368 patients) published between 2002 and 2006 suggest a sensitivity of 79% and a specificity of 97% in the trauma setting. In the two largest investigations, no false negative results were observed. Further research is needed to determine the efficacy of CTA for disclosing BCVI, and to evaluate the potential benefits to patients.     

Age-related changes in the response of rat adipocytes to insulin: evidence for a critical role for inositol phosphoglycans and cAMP

Adipose tissue plays a pivotal role in ageing and longevity; many studies, both human and animal, have focussed on the effects of food limitation. Here we present a new model based on striking differences between two ‘normal’ inbred strains of albino Wistar rats the Charles River (CR) and Harlan Olac (HO) that have marked differences in age-related accumulation of fat and insulin-stimulated rates of glucose incorporation into lipid in the epididymal fat pads (EFP). The incorporation [U-¹⁴C]glucose into lipid by adipocytes showed that the CR group had a twofold higher basal rate of lipogenesis and a greater response to insulin in vitro, exceptionally, adipocytes from CR group maintained the high response to insulin to late adulthood while retaining the lower EFP weight/100 g body weight. Inositol phosphoglycan A-type (IPG-A), a putative insulin second messenger, was 3.5-fold higher and cAMP significantly lower per EFP in the CR versus HO groups. Plasma insulin levels were similar and plasma leptin higher in CR versus HO groups. The anomaly of a higher rate of lipogenesis and response to insulin and lower EFP weight in the CR group is interpreted as the resultant effect of a faster turnover of lipid and stimulating effect of leptin in raising fatty acid oxidation by muscle, potentially key to the lower accumulation of visceral fat. The metabolic profile of the CR strain provides a template that could be central to therapies that may lead to the lowering of both adipose and non-adipocyte lipid accumulation in humans in ageing.     

Differential oxidation of two thiophene-containing regioisomers to reactive metabolites by cytochrome P450 2C9

The uricosuric diuretic agent tienilic acid (TA) is a thiophene-containing compound that is metabolized by P450 2C9 to 5-OH-TA. A reactive metabolite of TA also forms a covalent adduct to P450 2C9 that inactivates the enzyme and initiates immune-mediated hepatic injury in humans, purportedly through a thiophene-S-oxide intermediate. The 3-thenoyl regioisomer of TA, tienilic acid isomer (TAI), is chemically very similar and is reported to be oxidized by P450 2C9 to a thiophene-S-oxide, yet it is not a mechanism-based inactivator (MBI) of P450 2C9 and is reported to be an intrinsic hepatotoxin in rats. The goal of the work presented in this article was to identify the reactive metabolites of TA and TAI by the characterization of products derived from P450 2C9-mediated oxidation. In addition, in silico approaches were used to better understand both the mechanisms of oxidation of TA and TAI and/or the structural rearrangements of oxidized thiophene compounds. Incubation of TA with P450 2C9 and NADPH yielded the well-characterized 5-OH-TA metabolite as the major product. However, contrary to previous reports, it was found that TAI was oxidized to two different types of reactive intermediates that ultimately lead to two types of products, a pair of hydroxythiophene/thiolactone tautomers and an S-oxide dimer. Both TA and TAI incorporated 1?O from 1?O? into their respective hydroxythiophene/thiolactone metabolites indicating that these products are derived from an arene oxide pathway. Intrinsic reaction coordinate calculations of the rearrangement reactions of the model compound 2-acetylthiophene-S-oxide showed that a 1,5-oxygen migration mechanism is energetically unfavorable and does not yield the 5-OH product but instead yields a six-membered oxathiine ring. Therefore, arene oxide formation and subsequent NIH-shift rearrangement remains the favored mechanism for formation of 5-OH-TA. This also implicates the arene oxide as the initiating factor in TA induced liver injury via covalent modification of P450 2C9. Finally, in silico modeling of P450 2C9 active site ligand interactions with TA using the catalytically active iron-oxo species revealed significant differences in the orientations of TA and TAI in the active site, which correlated well with experimental results showing that TA was oxidized only to a ring carbon hydroxylated product, whereas TAI formed both ring carbon hydroxylated products and an S-oxide.     

CD40‐activated B cells can be generated in high number and purity in cancer patients: analysis of immunogenicity and homing potential

Cellular adjuvants such as dendritic cells (DC) are in the focus of tumour immunotherapy. In DC‐vaccine trials, induction of tumour antigen‐specific immunity is observed frequently and well‐documented clinical responses have been reported. However, the overall response rate is less than 3%, therefore alternative strategies are being investigated. CD40‐activated B cells (CD40‐B) have been characterized previously as an interesting alternative because they present antigen efficiently and can be expanded by several logs from small amounts of peripheral blood. To determine the central technical challenges of cell‐based vaccines we performed a single‐patient analysis of 502 patients from DC‐based tumour vaccine trials and identified at least three factors contributing to their limited efficiency: (1) lack of cell numbers; (2) lack of documented purity thus high contamination of bystander cells; and (3) lack of quality control and thus heterogeneous or unknown expression of important surface molecules such as major histocompatibility complex (MHC) and chemokine receptors. Based on these findings we re‐evaluated the CD40‐B approach in cancer patients. Here, we show that proliferation of B cells from cancer patients is equivalent to that observed in healthy donors. Purity is always > 90% after 2 weeks and remains stable for several weeks. They have comparable antigen‐presenting capability determined phenotypically and by allogeneic mixed lymphocyte reaction. Expression of CCR7 and CD62L was detected in all samples and B cells migrated towards the relevant homing chemokines. Taken together, CD40‐B cells from cancer patients can be expanded in virtually unlimited numbers at high purity and full function concerning antigen‐presentation and migratory properties.     

Behandlung nicht mit zystischer Fibrose assoziierter Bronchiektasen (Non-CF-Bronchiektasen)

Bronchiectasis has become more rarely because of the development of antibiotic therapy and vaccination. At present the great majority of bronchiectasis is more likely caused by congenital disorders than by infective reasons. Therapeutic strategies based on the experiences from cystic fibrosis and chronic obstructive pulmonary disease are not always conferrable to patients suffering from bronchiectasis. There are not enough controlled studies to give evidence-based recommendations in the treatment of bronchiectasis, which are not associated with cystic fibrosis. Goals in the treatment are improvement of the mucociliar clearance, the therapy of infections and treatment of inflammation. Currently several agents are under examination. To improve the prognosis and therapy options it would be reasonable to build up a national register for patients with bronchiectasis.     

GBT021601 improves red blood cell health and the pathophysiology of sickle cell disease in a murine model

The pathophysiologic mechanism of sickle cell disease (SCD) involves polymerization of deoxygenated haemoglobin S (HbS), leading to red blood cell (RBC) sickling, decreased RBC deformability, microvascular obstruction, haemolysis, anaemia and downstream clinical complications. Pharmacological increase in the concentration of oxygenated HbS in RBCs has been shown to be a novel approach to inhibit HbS polymerization and reduce RBC sickling and haemolysis. We report that GBT021601, a small molecule that increases HbS-oxygen affinity, inhibits HbS polymerization and prevents RBC sickling in blood from patients with SCD. Moreover, in a murine model of SCD (SS mice), GBT021601 reduces RBC sickling, improves RBC deformability, prolongs RBC half-life and restores haemoglobin levels to the normal range, while improving oxygen delivery and increasing tolerance to severe hypoxia. Notably, oral dosing of GBT021601 in animals results in higher levels of Hb occupancy than voxelotor and suggests the feasibility of once-daily dosing in humans. In summary, GBT021601 improves RBC health and normalizes haemoglobin in SS mice, suggesting that it may be useful for the treatment of SCD. These data are being used as a foundation for clinical research and development of GBT021601.     

Optimization of contrast agent volume for helical CT in the diagnostic assessment of patients with severe and multiple injuries.

PURPOSE: The aim of this study was to determine the optimal amount of contrast agent for helical CT of the trunk during primary radiologic evaluation of polytraumatized patients. METHOD: Eighty-three patients with severe and multiple injuries (mean age 36.2 years) underwent standardized helical CT examination with administration of a single contrast agent bolus of iohexol (Accupaque 300) at volumes of 120, 150, and 180 ml. Image quality was estimated by two blinded radiologists using a visual analogue scale. Analysis further included density measurements in regions of interest (ROIs) placed in the ascending, descending, and abdominal aorta as well as in the liver and spleen. RESULTS: The qualitative rating of the contrast agent effect after administration of 150 and 180 ml was significantly better compared with 120 ml [odds ratio (OR) 12.05, 95% confidence interval (CI) 3.50-41.52 and OR 12.14, 95% CI 3.36-43.85, respectively]. A dose increase from 120 to 150 ml resulted in a significantly better enhancement of the abdominal aorta (p = 0.006). The highest dose of 180 ml was not associated with a diagnostic gain in the other ROIs. CONCLUSION: We recommend administration of 150 ml of iohexol as the optimal amount of contrast material for single phase bolus administration in emergency helical CT examination of the chest and abdomen.