In Uruguay, an ethic of care for the dying

The ethics of care for the terminally ill in Uruguay spring from deeply rooted attitudes which invest the physician with sole power to determine the course of treatment, even to prescribe painkilling drugs and to hasten death actively or passively, without collaboration from the patient's family. The paternal function attributed to the physician has not been questioned by any systematic reflection on biomedical ethics. However, conscientious physicians are interested in these issues and are well informed about the concerns of their foreign colleagues.     

Epidermal growth factor receptor regulates normal urothelial regeneration

Members of the epidermal growth factor (EGF) family and their receptors are involved in many cellular processes, including proliferation, migration, and differentiation. We have previously reported that these growth factors are expressed and have specific regulatory functions in an organ-like culture model of normal human urothelial cells. Here, we used this model to investigate the involvement of EGF receptor (EGFR) in human urothelial regeneration. Three 4-mm-diameter damaged areas were made in confluent normal human urothelial cell cultures with a biopsy punch. Regeneration was measured, on fixed stained cultures, with an image analyzer, at 4, 24, and 48 hours after injury. Cell proliferation was assessed by 5-bromo-2-deoxyuridine incorporation. To identify EGF family factors potentially involved in the healing process, we studied the effect of these factors on damaged confluent cultures and the level of expression of mRNAs extracted from these cultures. EGFR inhibition of the proliferation and migration of urothelial cells was tested with (1). a specific tyrosine kinase inhibitor (AG1478) and (2). a blocking anti-EGFR antibody (LA22). Exogenously added amphiregulin, EGF, transforming growth factor-alpha and heparin-binding EGF (HB-EGF) stimulated urothelial regeneration. The damaged areas were repaired by regrowth within 48 hours. Both AG1478 and LA22 inhibited the repair (by 50% and 30%, respectively), as well as proliferation and migration. This regeneration was accompanied by increased HB-EGF mRNA expression in cultures of cells from four of six subjects, but no corresponding change in EGFR protein level was observed. These results indicate that the EGFR signaling pathway is involved in urothelial regeneration. Our data support an autocrine role of HB-EGF in this process and suggest that the EGFR pathway is a potential therapeutic target for modulating urothelial cell proliferation.     

Hemocompatibility of medical connectors with biopassive or bioactive surface coatings

Although medical connectors compose very small parts of the extracorporeal circulation (ECC) system they represent a critical localization where early thromboembolic processes can manifest. In the present study we modified an in vitro closed-loop model with fresh human whole blood for the preclinical evaluation of the hemocompatibility of three types of medical connectors: non-coated (control); with silicone-, and heparin-coating. Each single loop consists of five polycarbonate connectors joined together by five pieces of silicone tubes. Thrombin-antithrombin-III, beta-thromboglobulin (beta-TG), PMN-Elastase, terminal complement complex, CD 11b expression, and surface-absorbed fibrinogen were measured. After 1 and 2 h recirculation, platelet loss, release of beta-TG, and adsorption of fibrinogen were significantly higher (p<0.05) within the non-coated connectors compared to the silicone- and heparin-coated groups. Following this experiment, the connectors were filled again with fresh heparinized whole blood from the same donor to evaluate the influence of prior blood contact. Here, the activation of platelets and coagulation was dependent on the duration of the blood preincubation period. Probably, the coated surfaces possess a reduced, or selective adsorption of plasma proteins, which in turn leads to a faster creation of a blood-friendly secondary superficial membrane, and prevents a further denaturation and hence activation of the adsorbed proteins.     

Cranial sutures and craniometric points detected on MRI

The main goal of the study was to determine on MRI the cranial sutures, the craniometric points and craniometric measurements, and to correlate these results with classical anthropometric measurements. For this purpose, we reviewed 150 cerebral MRI examinations considered as normal (Caucasian population aged 2049 years). For each examination we individualized 11 craniometric landmarks (Glabella, Bregma, Lambda, Opisthocranion, Opisthion, Basion, Inion, Porion, Infra-orbital, Eurion) and three measurements. Measurements were also calculated independently on 498 dry crania (Microscribe 3-DX digitizer). To validate the MRI procedure, we measured four dry crania by MRI and with compass or digital caliper gauges. Cranial sutures always appeared without signal (black), whatever the MRI sequence used, and they are better visualized with a 5 mm slice thickness (compact bone overlapping). Slice dynamic analysis and multiplanar reformatting allowed the detection of all craniometric points, some of these being more difficult to detect than others (Porion, Infra-orbital). The measurements determined by these points were as follows: VertexBasion height=135.66±6.56 mm; EurionEurion width=141.17±5.19 mm; GlabellaOpisthocranion length=181.94±6.40 mm. On the midline T1-weighted sagittal image, all median craniometric landmarks can be individualized and the GlabellaOpisthocranion length, VertexBasion height and parenchyma indices can be calculated. Craniometric points and measurements between these points can be estimated with a standard cerebral MRI examination, with results that are similar to anthropometric data.     

Relating cell and tissue mechanics: Implications and applications

The Differential Adhesion Hypothesis (DAH) posits that differences in adhesion provide the driving force for morphogenetic processes. A manifestation of differential adhesion is tissue liquidity and a measure for it is tissue surface tension. In terms of this property, DAH correctly predicts global developmental tissue patterns. However, it provides little information on how these patterns arise from the movement and shape changes of cells. We provide strong qualitative and quantitative support for tissue liquidity both in true developmental context and in vitro assays. We follow the movement and characteristic shape changes of individual cells in the course of specific tissue rearrangements leading to liquid-like configurations. Finally, we relate the measurable tissue-liquid properties to molecular entities, whose direct determination under realistic three-dimensional culture conditions is not possible. Our findings confirm the usefulness of tissue liquidity and provide the scientific underpinning for a novel tissue engineering technology. Developmental Dynamics 237:2438-2449, 2008. (c) 2008 Wiley-Liss, Inc.     

Intensivtherapie und Prognose des akuten Kreislaufstillstandes

Zusammenfassung Anhand von 115 im Zeitraum von 2 1/2 Jahren beobachteten Episoden von akutem Kreislaufstillstand bei 90 Patienten werden die Kriterien und Behandlungserfolge der Intensivtherapie des akuten Kreislaufstillstandes dargelegt. Während die Basis-Therapie mit externer Herzmassage, künstlicher Beatmung, Natriumbicarbonatinfusionen und elektrischer Defibrillation eindeutig feststeht, bietet die medikamentöse Führung noch immer Probleme. Zehn eigene Verlaufsbeobachtungen zeigen, daß betaadrenergische Sympathomimetica auch bei bestimmten Fällen tachysystolischer Herzrhythmusstörungen günstig wirken. Der Versuch der elektrischen Stimulation des Herzens mit einer extern angelegten epikardialen Schrittmachersonde bei akutem Kreislaufstillstand durch Asystolie oder Bradykardie führte in keinem von 8 Fällen zum Erfolg. Für die Früh- und Spätprognose sind bei adäquater Therapie in erster Linie Art und Schwere des Grundleidens sowie der morphologisch funktionelle Zustand lebenswichtiger Organe bei Einsetzen des plötzlichen Kreislaufstillstandes verantwortlich. Bei Kammerflimmern und Kammerflattern waren deutlich bessere Resultate zu erzielen, als beim akuten Kreislaufstillstand durch Asystolie. Von den 90 Patienten überlebten 12 länger als 4 Wochen (13%), 9 (10%) konnten entlassen werden und sind zum Zeitpunkt der Publikation ohne Nachwirkungen des akuten Kreislaufstillstandes. Es wird auf die Bedeutung einer lückenlosen Beobachtung Schwerstkranker und die Beachtung prämonitorischer Zeichen zur Verhütung des akuten Kreislaufstillstandes hingewiesen.

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Summary Criteria and therapeutic success of intensive therapy of cardiac arrests are evaluated in 115 episodes observed in 90 patients over a period of 2 1/22 years. While basic therapy with external cardiac massage, artificial ventilation, infusion of sodium bicarbonate and electrical defibrillation is generally agreed upon, the choice of further drugs is still problematic. It is shown in 10 cases that under certain conditions even in ventricular fibrillation and ventricular tachycardia beta-receptor stimulating drugs are indicated. Electrical stimulation of the heart by an external epicardial pacemaker was unsuccessful in all of eight cases with complete asystoly or bradycardia. Immediate and long term results of cardiac arrests with adequate treatment primarely depends on the extend of the underlying disease as well as on the condition of all vital organs at the time of onset of the arrest. Ventricular fibrillation and -flutter had a better prognosis than asystoly. Twelve out of the ninety patients lived longer than four weeks after the arrest (13%). Nine of these are still living at the time of publication with an observation time from three months to two years. The chance of improving therapeutic results with intensive care lie mainly in preventing rather than curing complications such as cardiac arrests.

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Professor Dr.H. E. Bock zum 65. Geburtstag.     

Immunization against avian proteins.

A study of sera of pigeon breeders showed a higher ratio of antibodies with an anti-P1 specificity in those who show clinical signs of allergic origin. By absorption of anti-P1 antibodies it was revealed that there exist in the cells, serum and excrement of pigeons, substances with antigenic properties similar to those of human P1 antigen. Pigeon breeders, and particularly those who show clinical signs of allergy, possess also other antibodies which precipitate specific antigens of pigeon serum.     

Linkage of the horizontally acquired ypm and pil genes in Yersinia pseudotuberculosis

The superantigen-encoding ypm gene and the pil gene cluster governing type IV pilus biogenesis have been laterally acquired by Yersinia pseudotuberculosis. PCR assays on 270 unrelated strains from various environmental and animal sources revealed a significant association of ypm and pil in isolates.     

Regulation of main olfactory bulb mitral cell excitability by metabotropic glutamate receptor mGluR1

In the rodent main olfactory bulb (MOB), mitral cells (MCs) express high levels of the group I metabotropic glutamate receptor (mGluR) subtype, mGluR1. The significance of this receptor in modulating MC excitability is unknown. We investigated the physiological role of mGluR1 in regulating MC activity in rat and mouse MOB slices. The selective group I agonist (RS)-3,5-dihydroxyphenylglycine (DHPG), but not group II or III agonists, induced potent, dose-dependent, and reversible depolarization and increased firing of MCs. These effects persisted in the presence of blockers of fast synaptic transmission, indicating that they are due to direct activation of mGluRs on MCs. Voltage-clamp recordings showed that DHPG elicited a voltage-dependent inward current consisting of multiple components sensitive to potassium and calcium channel blockade and intracellular calcium chelation. MC excitatory responses to DHPG were absent in mGluR1 knockout mice but persisted in mGluR5 knockout mice. Broad-spectrum LY341495, MCPG, as well as preferential mGluR1 LY367385 antagonists blocked the excitatory effects of DHPG and also potently modulated MC spontaneous and olfactory nerve-evoked excitability. mGluR antagonists altered spontaneous membrane potential bistability, increasing the duration of the up and down states. mGluR antagonists also substantially attenuated MC responses to sensory input, decreasing the probability and increasing the latency of olfactory nerve-evoked spikes. These findings suggest that endogenous glutamate tonically modulates MC excitability and responsiveness to olfactory nerve input, and hence the operation of the MOB circuitry, via activation of mGluR1.     

Use of tuf sequences for genus-specific PCR detection and phylogenetic analysis of 28 streptococcal species

A 761-bp portion of the tuf gene (encoding the elongation factor Tu) from 28 clinically relevant streptococcal species was obtained by sequencing amplicons generated using broad-range PCR primers. These tuf sequences were used to select Streptococcus-specific PCR primers and to perform phylogenetic analysis. The specificity of the PCR assay was verified using 102 different bacterial species, including the 28 streptococcal species. Genomic DNA purified from all streptococcal species was efficiently detected, whereas there was no amplification with DNA from 72 of the 74 nonstreptococcal bacterial species tested. There was cross-amplification with DNAs from Enterococcus durans and Lactococcus lactis. However, the 15 to 31% nucleotide sequence divergence in the 761-bp tuf portion of these two species compared to any streptococcal tuf sequence provides ample sequence divergence to allow the development of internal probes specific to streptococci. The Streptococcus-specific assay was highly sensitive for all 28 streptococcal species tested (i.e., detection limit of 1 to 10 genome copies per PCR). The tuf sequence data was also used to perform extensive phylogenetic analysis, which was generally in agreement with phylogeny determined on the basis of 16S rRNA gene data. However, the tuf gene provided a better discrimination at the streptococcal species level that should be particularly useful for the identification of very closely related species. In conclusion, tuf appears more suitable than the 16S ribosomal RNA gene for the development of diagnostic assays for the detection and identification of streptococcal species because of its higher level of species-specific genetic divergence.