The ability of heat stress and metabolic preconditioning to protect primary rat cardiac myocytes

Primary rat cardiocytes were subjected to either thermal preconditioning for 30 min at 43°C or 20 min metabolic preconditioning (10 mM deoxyglucose, 20 mM lactate, pH 6.5). Eighteen hours later cells were analysed either for hsp 70i expression or subjected to a subsequent lethal heat stress or simulated ischaemia (10 mM deoxyglucose, 20 mM lactate, 0.75 mM sodium dithionite, 12 mM potassium chloride, pH 6.5) for 2 hours and assessed for survival by trypan blue exclusion.Hsp 70i was induced over 100 fold by thermal preconditioning and 30 fold by metabolic preconditioning (p<0.001, p<0.05), hsp 90 was induced 2.71 fold and 2.24 fold (p<0.001, p<0.001) by thermal and metabolic preconditioning respectively, while hsp 60 was not induced by either treatment. Preconditioned cultures had improved survival against subsequent lethal heat stress or simulated ischaemia: Thermal preconditioning reduced death from 69.22% to 52.46% upon subsequent lethal heat stress and from 49.13% to 36.66% upon subsequent lethal simulated ischaemia. Metabolic preconditioning reduced cell death from 51.29% to 33.8% against subsequent lethal heat stress, and from 69.09% to 55.61% upon subsequent lethal simulated ischaemia. A second marker of cell death, the release of lactate dehydrogenase activity into the culture media, was reduced to 65% and 60% of control values for thermally preconditioned cells subjected to lethal heat or lethal simulated ischaemia respectively. Metabolically preconditioned cells demonstrated lactate dehydrogenase activity of 59% and 51% that of control values, when subjected to lethal heat or lethal simulated ischaemia respectively.Abbreviations hsp heat stress protein - hsp 70i inducible 70 kDa heat stress protein - LDH lactate dehydrogenase - PBS phosphate buffered saline     

Estimation of the malignant potential of gastrointestinal stromal tumors: the value of contrast-enhanced coded phase-inversion harmonics US

Background Recently, contrast agents for ultrasonography (US) such as Levovist have been introduced for routine clinical use. The contrast-enhanced US with Levovist permits evaluation of the intratumoral vascularity of hepatic and pancreatic tumors and is useful for their differential diagnosis. The purpose of the present study was to assess tumor vessels and the parenchymal flow of gastrointestinal stromal tumors (GISTs) by contrast-enhanced coded phase-inversion harmonic US and to evaluate whether vascularity is related to the malignant grade of the GISTs.Methods Thirteen patients with GISTs were included in the present study. Tumors were observed in a real-time fashion of contrast-enhanced coded phase-inversion harmonic US after the injection of Levovist (400mg/ml). The vascular patterns were compared with tumor size, histological diagnosis, KIT mutations, and clinical findings such as metastasis.Results The contrast-enhanced US images of the GISTs were classified into two types according to the blood flow area of the tumors as seen by real-time continuous imaging of the tumor vessels. The image pattern Poor represented vessels flowing only in the peripheral part of the tumor, and Rich represented abundant vessels flowing from the periphery to the central part of the tumor. According to the contrast-enhanced US images, five GISTs were classified as Poor and the others as Rich. Based on the final diagnosis, all tumors with Poor images were determined to be benign GISTs, and the rest tumors except one with Rich images were determined to be malignant GISTs.Conclusions Contrast-enhanced US image is more closely correlated with the final diagnosis than the histological findings.     

The establishment of a polyposis register

Guidelines are presented for the establishment of a regional or national register of patients with familial adenomatous polyposis. The detailed recommendations are based on the work in committees of the Leeds Castle Polyposis Group and the EuroFAP. The aims of national and regional polyposis registers are discussed, and the stages of development of a register are reviewed: Ascertainment of probands, construction of pedigrees, identification of family members at risk, and screening of members at risk. The problem of data confidentiality is discussed.

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Résumé Les consignes sont présentées pour l'établissement d'un registre Régional ou National des patients avec polypose adénomateuse familiale. Les recommandations détaillées sont basées sur les travaux des comités du Leeds Castle Polyposis Group et de EuroFAP. Les buts des registres Régionaux ou Nationaux des polyposes sont discutés et les stades de développement du registre sont passés en revu: l'assurance de probande, la construction de l'arbre généalogique, l'identification des familles des membres à risques et l'étude des membres à risques. Le problème de la confidentialité des faits est discuté.

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This paper is based on a report from committees of the Leeds Castle Polyposis Group and the EuroFAP.     

Ileocolitis: The importance of a more precise definition

Summary We have reviewed our experience with 22 patients with nonspecific inflammatory involvement of the small and large bowel. Patients with this combination of lesions usually have long-standing and intractable illnesses, with many complications, frequently resulting in multiple operations. In long-term follow-up we found a mortality of 23%.A particularly troublesome combination is granulomatous disease of the small bowel accompanying or followed by ulcerative colitis. Some suggested guidelines as to therapy in the management of these difficult cases have been presented.The term ileocolitis should be qualified, whenever possible, by the more specific pathologic terms granulomatous or ulcerative as an aid in planning therapy and in making a prognosis. In the occasional case with both of these characteristics, we subscribe to the term mixed enterocolitis.     

Interstitial pH value in the myocardium as indicator of ischemic stress of cardioplegically arrested hearts

Summary The influence of different cardioplegic methods (Bretschneider, Cardiac Surgery in Hamburg, Kirklin, St. Thomas' Hospital in London) on the progressive myocardial acidosis during global ischemia of up to 24 hours' duration was investigated in arrested, nonperfused canine hearts (n=36). The increasing acidosis of the ischemic heart muscle was continuously measured and registered with the aid of interstitial pH measurement (pH_i) using glass implant electrodes. With various buffered and unbuffered cardioplegic solutions there were relatively wide fluctuations of the pH_i courses in the myocardial extracellular space. The critical interstitial pH values (pH_crit) relative to a defined myocardial ATP concentration (4 mol/g ww) vary between 6.10 (Bretschneider) and 5.6 (Kirklin) (p<0.001). These striking results cannot be attributed solely to different total myocardial lactate concentrations, but variations in a cellular and an extracellular factor must also be taken into account.Measurement of interstitial pH for intraoperative monitoring of myocardial ischemic stress can therefore be used only within limits and with knowledge of the specific protective procedure applied. The method continues an important tool in basic investigations of partial or global ischemia of organs, particularly as regards the spectrum of metabolic, morphological and functional parameters.Dedicated to Prof. Dr. J. Koncz on the occasion of his 65th birthday     

Experimentelle Untersuchungen zur Frage der Seltenheit von Geschwulstmetastasen in der Skeletmuskulatur

Summary Authors injected a suspension of Brown-Pearce cancer cells into the left ventricle of 10 rabbits. The venous blood of the lower extremities of these first rabbits was injected into the ear vein of 20 intact second rabbits. In the surviving 7 first rabbits metastases in the skeletal muscles frequently developed. The majority of second rabbits developed tumors indicating a transmuscular passage of tumour cells emboli in the first rabbits. Transmuscular passage of tumour cell was found not only immediately after the injection of the tumour cells, but also three minutes later.These facts contradict the rather speculative theory according to which the skeletal muscle is an unsuitable soil for the development of secondary tumours.     

Prognostic relevance of histological findings on bone marrow biopsy in myelodysplastic syndromes

Summary Bone marrow biopsy (BMB) has aroused growing interest as a possible aid in the diagnostic and prognostic evaluation of myelodysplastic syndromes (MDS). Previous reports have pointed out that MDS patients with blastic aggregates or severe bone marrow (BM) fibrosis are characterized by a worse clinical outcome. BMBs of 106 MDS patients were retrospectively reviewed, and relationships among the different histological parameters as well as clinicopathological correlations were looked for. Three patterns of BM blastic infiltration (diffuse, cluster, and large) were recognized. Overt leukemic transformation and overall survival were selected as prognostic end points. BM infiltration was diffuse in 18, cluster in 48, and large in 40 cases. RAEB-t patients accounted for about half of the large cases, and none had a diffuse pattern (p<0.01). Nineteen patients showed extensive BM fibrosis; most of them were characterized by cluster blastic infiltration and megakaryocyte hyperplasia. Leukemic transformation occurred in 67% of large cases (p<0.001) and in none of the cluster cases with severe BM fibrosis (p<0.01); however, survival was equally poor in these two groups because of early leukemic transformation (large cases) and BM failure (cluster cases). The FAB classification did not significantly correlate with prognosis. Patients with cluster BM infiltration and severe fibrosis can be regarded as a true separate MDS subset characterized by unique clinicopathological and prognostic features. Because of the subacute clinical behavior of most cases, and the poor performance status of many elderly patients, there is still controversy as to the best therapeutic approach in MDS. Histological analysis allowed two groups of MDS patients to be identified, both characterized by poor life expectancy, who could benefit from early aggressive chemotherapy.     

The efficacy of neuraminidase treatment in studies on red cell aging

Summary An in vitro system was developed to test the propensity of rat erythrocytes (RBC) toward phagocytosis by homologous peritoneal macrophages after in vivo aging or after in vitro surface modification. Old RBC obtained from erythrocyte populations separated into fractions on the basis of their density were found to be phagocytosed to a significantly greater extent than cells obtained from young cell fractions. Neuraminidase treatment of RBC resulted in extensive phagocytosis in comparison to control cells representing the whole population or old cells. Galactose oxidase treatment of neuraminidase treated RBC afforded no protection from uptake by macrophages. In contrast, treatment of RBC with neuraminidase immobilized on Sepharose 4B, leading to removal of up to 25% of the total membrane sialic acid, did not lead to phagocytosis above control values. These results indicate that extreme caution is necessary in the interpretation of experiments in which neuraminidase is utilized to produce simulated aged RBC.     

Use of cultured human tissues and cells in carcinogenesis research

Summary A variety of approaches are used to study carcinogenesis. Recent advances in techniques for culture of human tissues and cells have provided additional experimental systems of study the process of carcinogenesis and the genetics of cancer.The Journal of Cancer Research and Clinical Oncology publishes in loose succession Editorials and Guest Editorials on current and/or controversial problems in experimental and clinical oncology. These contributions represent exclusively the personal opinion of the author.     

Cytopathic Hypoxia in Sepsis: An Overview

Septic shock in humans is characterized by a hyperdynamic circulation, elevated systemic mixed-venous O2 content and lactic acidosis. An apparent defect in O2 extraction along with increased anaerobic mechanism has prompted investigation of the role of hypoxia in the production of organ dysfunction and death. Although classic hypoxia, i.e. anemic, hypoxic, and stagnant hypoxia may be present at the level of the macro- or microcirculation, there is increasing evidence for cytopathic hypoxia, failure of oxygen utilization in the presence of adequate tissue pO2. The process of cellular energy production via oxidative phosphorylation is discussed, and the general mechanisms through which it may be perturbed are reviewed: 1) failure to deliver adequate substrate to the electron transport chain, 2) dysfunction of the electron transport chain, 3) failure of proper coupling between electron flux and ATP synthesis, (4) failure to deliver adequate O2 to the electron transport chain. The first three mechanisms define cytotopathic hypoxia and are discussed in-depth, with reference to possible relevance in human sepsis. Synergy between classic hypoxia and cytopathic hypoxia is hypothesized. The expanding role of nitric oxide and reactive nitrogen intermediates in this process is highlighted.     

Binding of lectins to “young” and “old” human erythrocytes

Summary Old human erythrocytes showed a 21.2% decrease in cell surface area and a 2% decrease in the number of WGA receptor sites, but a 27% increase in the distribution density of the WGA (lectin) receptor site, when compared with young human erythrocytes. For a list of lectin abbreviations, see Materials and methods). Both young and old erythrocytes exhibited very weak binding activity for ¹²⁵I-labeled PNA, but there was no difference in binding activity for PNA between young erythrocytes and old ones. Compared with young erythrocytes, decreases in the number and distribution density of receptor sites for five lectins including LPA, Con A, RCA-II, SBA and BPA on the cell surface were observed in aged erythrocytes. Old erythrocytes also showed a decrease in the number of PHA-E receptor sites, while the distribution density of the same receptor site remained unchanged. In view of these and other observations, it is thought that human erythrocyte aging is accompanied by elimination of some glycoconjugates which have affinity for six lectins, LPA, Con A, RCA-II, PHA-E, SBA and BPA, whereas no WGA receptor-containing glycoconjugates are released from erythrocyte membranes. Elimination of the glycoconjugates results in shrinkage of erythrocytes to reduce their cell surface areas.     

Insulin and growth hormone responses to glucose loading in treated acromegalics

Summary The insulin and growth hormone responses to oral glucose load (100 g) in 23 acronaegalic patients, previously treated by external irradiation, are described. Based on current clinical findings, sixteen acromegalics were considered inactive and seven patients active. Two of the latter were treated diabetics. Thirteen healthy, non-obese subjects formed the control group. Five inactive acromegalics had blood glucose values exceeding the upper limits of normal after the glucose load. Both the inactive and active acromegalics had hyperinsulinemia in the fasting state and after glucose load. The observation of an exaggerated serum insulin response to glucose load in clinically inactive acromegalics suggested that chronically elevated levels of circulating growth hormone may have led to permanent changes in the responsiveness of the pancreatic islets to glucose stimulation. The mean fasting value of serum growth hormone was about the same in the controls and the inactive acromegalics; the latter did not show suppression in serum growth hormone levels after glucose load.Research Fellow, Medical Research Council of Canada     

Endoscopic therapy of the “sump” syndrome

Choledochoduodenostomy may be followed by the sump syndrome. This previously required surgical intervention. We describe the successful application of endoscopic therapy in two patients with this syndrome.     

Susceptibility and tolerance of β-lactamase-producing,methicillin-sensitive strains of Staphylococcus aureus towards seven broad-spectrum penicillins

Summary The activity of penicillin G, ampicillin, carbenicillin, ticarcillin, azlocillin, mezlocillin and piperacillin against 102 -lactamase-producing, methicillin-sensitive strains ofStaphylococcus aureus was determined by agar dilution (method A) and broth microdilution (method B) techniques. By NCCLS breakpoint criteria, 4% of the strains were sensitive to penicillin and ampicillin, and almost 100% were sensitive to the other drugs when method A was used. Results with method B were only significantly lower as far as the cumulative percentage of strains sensitive to azlocillin, mezlocillin and piperacillin was concerned (63–71%). Bactericidal effects at sensitive levels were observed in 0–2% (penicillin, ampicillin), 31–35% (carbenicillin, ticarcillin) and 10–14% (azlocillin, mezlocillin, piperacillin). While differences in MIC and MBC levels ranged from 0 to 8 dilution steps, tolerance (a >32-fold difference) was seen in at least 9–22% of all strains (depending on the drug tested); experimental limitations, however, excluded a determination of tolerance in all our strains. In a semi-quantitative nitrocefin assay, strong -lactamase production was correlated to high MIC and/or MBC levels.

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Sensibilität und Toleranz Betalaktamase-produzierender, Methicillin-sensibler Stämme von Staphylococcus aureus gegenüber sieben Breitspektrumpenicillinen

Zusammenfassung Die Aktivitäten von Penicillin G, Ampicillin, Carbenicillin, Ticarcillin, Azlocillin, Mezlocillin und Piperacillin gegen 102 -Lactamase-produzierende, Methicillin-sensible Stämme vonStaphylococcus aureus wurden mit Hilfe einer Agardilution (A) und Bouillon-Mikrodilution (B) bestimmt. Unter Zugrundelegung der NCCLS-Kriterien erwiesen sich bei Verwendung der Methode A 4% der Stämme als sensibel gegen Penicillin und Ampicillin und fast 100% sensibel gegen die anderen Penicilline. Mit Methode B ergaben sich signifikante Differenzen gegenüber A lediglich bei Azlocillin, Mezlocillin und Piperacillin (63–71% Sensibilität). Bakterizidie-Effekte im sensiblen Bereich ergaben sich bei 0–2% (Penicillin und Ampicillin), 31–35% (Carbenicillin und Ticarcillin) bzw. 10–14% (Azolocillin, Mezlocillin, Piperacillin). Differenzen zwischen MHK und MBK reichten von 0 bis 8 Verdünnungsstufen; und Toleranz (MBK> 32 MHK) wurde bei mindestens 9–22% der Stämme (je nach Antibiotikum) gesehen. Limitationen im Experiment ließen jedoch nicht bei allen Stämmen Auswertung auf Toleranz zu. Bei Verwendung einer semiquantitativen Nitrocefin-Bestimmungsmethode zeigte sich eine Korrelation zwischen starker -Laktamase-Produktion und hohen MHK- und/oder MBK-Werten.

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This paper is dedicated to Prof.Walter Siegenthaler on the occasion of his 60th birthday.     

Follow-up of “non-diabetic” relatives of diabetics by retesting oral glucose tolerance after 5 years

Summary Of 743 first degree relatives of diabetics in whom oral glucose tolerance tests had been performed in 1967 488 were re-tested in 1972. Among the original normals (n = 353) 17.6% had developed a subclinical and 1.3% an overt diabetes within 5 years. The original subclinical diabetics (n = 118) showed a remission to normal in 35.6% and a progression to overt diabetes in 13.6%. 3 out of the 17 formerly overt diabetics were found to be normal after 5 years and 3 were subclinical diabetics. Thus the performance of an oral glucose tolerance test is of limited prognostic value in the individual case. In both studies a higher prevalence of abnormal test results occurred in the older age groups and in overweight subjects. Remission or deterioration did not depend, however, on age or on weight changes. The frequency of abnormal tests was higher in males than in females, but the tendency towards the development of diabetes was more pronounced in females, in accordance with a previous observation of a higher age dependance of glucose tolerance in females.     

Heat stress increases cardiac HSP72i but fails to reduce myocardial infarct size in rabbits 24 hours later

Recent reports suggest that delayed myocardial protection (second window of preconditioning) occurs 24 hours after brief ischemic or thermal stress. In order to test this hypothesis, we subjected New Zealand White rabbits to a heating regimen (42°C for 15–20 minutes). Twenty four hours later, the effect of heat stress on infarct size was determined by conducting a 30 minute ischemia/3 hour reperfusion protocol. In a separate group of rabbits, Western blot analysis was used to verify that the heating regimen increased expression of HSP72i. The size of the region at risk was delineated by infusion of Unisperse blue and infarcted myocardium was identified by incubation of left ventricular slices in triphenyl tetrazolium chloride. In contrast to expectations, induction of HSP72i with thermal stress was not effective in limiting infarct size in rabbits 24 hours later, calling into question the concept that heat stress induces delayed or second window myocardial protection.     

Überlegungen und Hochrechnungen zur Epidemiologie des „Acquired Immunodeficiency Syndrome“ in der Bundesrepublik Deutschland

Zusammenfassung Aufgeschlüsselt nach Bevölkerungskollektiven mit unterschiedlichen sexuellen Verhaltensweisen wird anhand der bisher bekannten bzw. größenordnungsmäßig abgeschätzten epidemiologischen Daten des Acquired Immunodeficiency Syndrome eine Hochrechnung der Morbiditäts- und Mortalitätsziffern für die Bundesrepublik Deutschland mittels Computer-Simulation vorgenommen; danach könnten in den nächsten 15 Jahren bis ca. 41 000 Personen erkranken. Die Bedeutung der weiblichen Prostituierten und der männlichen Bisexuellen als Vektoren der Infektion aus den bisherigen Risikogruppen in Bevölkerungsteile mit ausschließlich heterosexuellen Verhaltensweisen, der Einfluß infektiologischer Schutzmaßnahmen sowie der Effekt sexueller Verhaltensänderungen breiter Bevölkerungsschichten kann mit diesem Programm ebenfalls untersucht werden.

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Considerations and simulations regarding the epidemiology of the acquired immunodeficiency syndrome in the federal republic of Germany

Summary A computer simulation of the morbidity and mortality rates of the Acquired Immunodeficiency Syndrome in the Federal Republic of Germany was performed. Since sexual intercourse is the main mode of transmission, the population was divided into six groups with different sexual behaviour. In several variations of the program it is demonstrated that during the next 15 years up to 41,000 AIDS patients can be expected. The importance of female prostitutes and male bisexuals as vectors who may transmit the infection into exclusively heterosexual groups is shown. Using this program, the effect of prophylaxis against infection as well as of changes of sexual behaviour within the population at large can also be examined.

     

The neo-synovial membrane of patients with active rheumatoid arthritis at resynovectomy

Summary Neo-synovial membranes, which formed after primary synovectomy in 21 patients with rheumatoid arthritis (RA), were studied at resynovectomy. The clinical, histomorphological, and immunohistological data were compared with data derived from primary synovial membranes from RA and osteoarthritis (OA) patients. The clinical data suggest a less active rheumatoid inflammatory response after synovectomy. Histomorphologicaly, the synovitis in resynovectomized neosynovial membranes of RA revealed no qualitative differences when compared with synovitis in the primary synovium. However, the degree of the inflammatory rection evaluated by the different parameters was found to be distinctly lower. The immunohistological data correlated with these findings.     

Chromosomenuntersuchung zur Charakterisierung eines Impftumors

Zusammenfassung Chromosomenzahl und marker-chromosomen eines Impftumors der Maus (Lymphoms) wurden bestimmt. Die Stammlinie mit 45 Chromosomen sowie zwei marker-chromosomen weisen eine auffallende Übereinstimmung mit Befunden an Ehrlich-Carcinomstämmen auf. Das zweischenkelige marker-chromosom tritt in zwei unterschiedlichen Varianten auf, ein weiteres weniger deutlich ausgeprägtes sehr langes telozentrisches Element wurde als drittes marker-chromosom dieses Tumors betrachtet.Unter gleichbleibenden Transplantationsbedingungen blieb der Karyotyp durch 14 Monate hindurch konstant. Auch durch hochdosierte Oestrogengaben an die tumortragenden Mäuse konnte nach 21 Passagen keine wesentliche Änderung des Karyotyps erzielt werden, sondern lediglich eine allmähliche Zunahme der Tumorzellen mit 46–49 Chromosomen, bei gleichbleibenden marker-chromosomen.

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Summary The chromosome number and marker-chromosomes of a transplantable tumor with of the mouse (lymphoma) were determined. The stemline 45 chromosomes and the two marker-chromosomes corresponded strikingly with results of studies on strains of Ehrlich carcinoma. The two-limbed marker-chromosome appeared in two different variations, and as a third marker-chromosome of this tumor a less striking but very long telocentric element was further observed. Throughout a 14 month period under constant conditions of transplantation the karyotype remained unchanged. Furthermore, no important changes of the karrotype could be obtained by high doses of estrogen in tumor-bearing mice after 21 passages. Only a gradual increase of tumor cells with 46–49 chromosomes occurred; the marker chromosomes remained constant.

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Mit 4 Textabbildungen     

The Utilization of Colon Transit Scintigraphy in the Diagnostic Algorithm for Patients with Chronic Constipation

We performed a retrospective utilization studycovering a four-year period to determine how physiciansfamiliar with colon transit scintigraphy (CTS) use it tomanage patients with chronic constipation. Following CTS, there was a change is bothfrequency and likelihood of diagnosis. The diagnosis waschanged in 51% of patients — 37% of thoseconsidered to have slow transit constipation (STC)before CTS, 43% with obstructed defecation, and 64% withfunctional bowel disease (FBD). CTS increased thediagnostic likelihood in all groups. Of patients withthe diagnosis of STC, 16% were considered almost certain before CTS while 83% wereconsidered almost certain after CTS. ForFBD comparable percentages were 13% and 62%. CTS mayplay a major role in the diagnostic work-up of patientswith chronic constipation, both altering diagnosis andincreasing diagnostic certainty.