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     

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.     

Extrarenal nephroblastomas

Summary The cases of extrarenal nephroblastoma published in the world literature have been tabulated and reviewed, and are discussed from a historical point of view.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     

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.     

Gastric “pseudolymphoma” with restricted light chain expression in a patient with obscure gastrointestinal blood loss

Summary We present a case report of a gastric pseudolymphoma (GL) that exhibits restricted light chain expression and, therefore, illustrates the dilemma encountered when histologically benign lesions have phenotypic abnormalities suggesting malignancy. For many years this lesion has been considered usually benign; however, recent reports such as this one demonstrate immunologic signs of monoclonality in an otherwise histologically benign lesion. This finding challenges our assumptions as to the nature of this lesion and our definition of cancer. The lesion also displays Castleman-like features (angiofollicular hyperplasia) as well as common inflammatory germinal centers. In addition we briefly review the literature.     

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.     

An auto-anti-B in an A1B person. Serological studies

Summary The serum of a patient (Mr. Lat) with the regular blood group A₁B contains an anti-B reacting with all cells having a B antigen except b_x and cis AB. The anti-B reacts at 4° C and occasionally at room temperature as shown by agglutination, absorption-elution and by thermo-dynamic assays. The antibody is regarded as an irregular autoantibody belonging to the group of the so called suppressed or latent antibodies.

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Zusammenfassung Das Serum eines Patienten (Mr. Lat) mit der normalen Blutgruppe A₁B enthält Anti-B, das mit allen Zellen reagiert, die ein B-Antigen aufweisen, ausgenommen B_x und cis AB. Das Anti-B reagiert bei 4° C und gelegentlich bei Zimmertemperatur, was durch Agglutination, Absorption-Elution und durch thermodynamische Unter-suchungen gezeigt wurde. Der Antikörper wird als irregulärer Autoantikörper angesehen, der zur Gruppe der supprimierten oder latenten Antikörper gehört.

     

Impairment of monocyte “lectin-like” receptor activity in Type 1 (insulin-dependent) diabetic patients

Summary In order to investigate whether the ability of peripheral blood monocytes to bind bacteria is impaired in diabetes, we studied carbohydrate-binding (lectin-like) receptors and the receptor for the Fc portion of immunoglobulin on monocytes from 25 male Type 1 (insulin-dependent) diabetic patients and 10 age-matched healthy control subjects. Peripheral blood monocytes from the diabetic patients expressed lower levels of lectin-like receptors compared to the control subjects, whereas the expression of the receptor for the Fc portion of immunoglobulin was similar in both populations. There was no correlation between the degree of lectin-like binding activity and plasma glucose concentration or glycaemic control. Recognition of unopsonized bacteria by the lectin-like receptor is impaired in Type 1 diabetes; this may affect the efficient elimination of potential pathogens.     

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.     

Slow metabolic deterioration towards diabetes in islet cell antibody positive patients with autoimmune polyendocrine disease

Summary We studied metabolic progression to IDDM in a cohort of adults who are ICA-positive and have associated autoimmune endocrine disease or circulating organ-specific autoantibodies (the Polyendocrine Study). Of the 186 individuals recruited 27 developed overt diabetes after a median follow-up of 4.5 years (range 0.4–12). Of these, eight patients did not require insulin treatment until at least 6 months after clinical diagnosis, with an interval of 1.8 years (1.2–5.7). An IVGTT was performed in 38 subjects and 23 had sequential studies. Of the initial 38 subjects six developed diabetes and only three showed a loss of FPIR to glucose (below the first percentile of a normal control group) before clinical onset of the disease. An additional three subjects showed a loss of the FPIR, and all still have normal glucose tolerance after median follow-up of 28 months (22–95). A whole or mixed pattern of islet cell staining was found in five of the six patients who developed diabetes and antibodies against an islet 37 k-antigen were detectable in four patients, all of whom required insulin soon after diagnosis. A beta-cell selective ICA staining pattern was seen in 14 of 17 subjects who did not develop diabetes and the mixed pattern in only three. None of this group had detectable 37k-antibodies. We conclude that metabolic deterioration is slow in polyendocrine patients, and that the IVGTT has less prognostic significance in this group than in first degree relatives of patients with IDDM. In contrast, the presence of the whole or mixed ICA staining pattern or of 37k-antibodies can identify a high risk of progression to IDDM within this polyendocrine population and may indicate the rate of metabolic deterioration.Abbreviations IDDM Insulin-dependent diabetes mellitus - ICA islet cell antibodies - IVGTT intravenous glucose tolerance test - FPIR first phase insulin response - OGTT oral glucose tolerance test - GAD glutamic acid decarboxylase - JDF Juvenile Diabetes Foundation     

Functional and surface characteristics of lymphocytes from patients with warm-antibody type autoimmunhemolytic anemia (AIHA)

Summary As part of an overall assessment of immunological function, several aspects of cellular immunity and circulating lymphocyte subpopulations were evaluated in a group of 10 patients with idiopathic autoimmunhemolytic anemia (AIHA). The absolute numbers of circulating T and B cells were reduced in the patient group compared to normals. A shift from corticosteroid-sensitive to corticosteroid-resistent and activated cells in the cytogram of clustered Fe-(III)-hydroxide-glucane saccharose labeled T lymphocytes was apparent. In vitro studies of cellular reactivity, as evaluated by PHA, ConA, PWM, antigens and allogeneic cell induced proliferation showed a blend of general or selective depression and sometimes a normal or increased activity with no definite correlation with both the number of circulating T cells and the extent of the hemolytic activity by the disease. The possible significance of the findings is discussed.This work was supported by the Minister für Wissenschaft und Forschung des Landes NRW II B8-FA 6438     

Where is the origin of the activator calcium in cardiac ventricular contraction?

Summary Under normal experimental conditions, the force of rested-state contractions (i.e., contractions after a rest period of 15 min or longer) of mammalian ventricular myocardium is insignificant. In Mg²⁺-free solution, in low sodium solution or in the presence of a cardioactive steroid, a strong early restedstate contraction develops without delay after stimulation, indicating the accumulation during rest of intracellularly stored activator calcium. By contrast, catecholamines cause a late rested-state contraction with a characteristic latent period of about 100 ms between stimulation and onset of contraction.Inhibition of the slow inward current by nifedipine has no influence on the contraction velocity of the early rested-state contraction, indicating that Ca²⁺ of the slow inward current is not involved in the calcium release mechanism of prefilled stores during excitation-contraction coupling. Nifedipine suppresses the late rested-state contraction in the presence of noradrenaline. In view of the constancy of the latent period, it is proposed that the activator calcium for the late rested-state contraction enters the cell with the slow inward current, is sequestered at first by uptake sites of the sarcoplasmic reticulum and subsequently released from its release sites as long as the cell is depolarized.The model of the different origin of activator calcium is discussed, in its implication for high-frequency contractions.     

Immunoglobulin production and secretion in bence jones protein myeloma and “nonsecretory” myeloma

Summary Using our electron microscopic method for polysome analysis and an immunofluorescent technique we studied Ig production and secretion by tumor cells in seven BJP myeloma patients and seven nonsecretory myeloma patients.In BJP myeloma Ig production and secretion is of three types: Type 1, only L-chains are synthesized and secreted; Type 2, the myeloma cells show fluorescence for H-chains, but upon polysome analysis there is no peak at polysomes corresponding to H-chain production; Type 3, the myeloma cells show fluorescence for H-chains, and polysome analysis shows two peaks corresponding to L- and H-chain production.Polysome analysis of nonsecretory myelomas show the presence of only very few membrane-bound polysomes and their distribution curves are entirely different from those of ordinary myeloma. Furthermore, the distribution patterns vary among seven cases. Results obtained by polysome analysis and immunofluorescent technique suggest that the nonsecretory myeloma could be divided into several subtypes.Supported in part by grants-in-aid for scientific Research (748103, 867050, 167092, 267108, 777124) from the Ministry of Education, Science and Culture of Japan     

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.     

Effect of mucosal thickening near gastric carcinoma on the endoscopic diagnosis of malignancy

Gastric mucosal thickening of variable degree occurs in the vicinity of gastric carcinomas and is possibly related to simultaneous tumor expression of epidermal growth factor and its receptor. Seventeen cases in which both endoscopic biopsy and subsequent resection for gastric carcinoma had been performed were studied to see if putative tumor-related mucosal thickening had an effect on endoscopic biopsy sensitivity. Biopsy fragment positivity rate was greater in cases with exophytic, protruding tumor masses (46.8± 8.5%) than in all other cases (17.0± 4.7%; P=0.02). Thickness of nontumorous mucosa adjacent to carcinomas did not significantly affect the biopsy fragment positivity rate in cases with exophytic masses (thin subgroup, 51.0±16.2%; thick subgroup, 44.0±10.8%; P=0.7) but did reduce the positivity rate significantly (P=0.05) in ulcerative or infiltrative tumors without exophytic components (thin subgroup, 23.3±4.1%; thick subgroup, 8.4±2.1%). This reduction in biopsy sensitivity related to mucosal thickening occurring adjacent to nonprotuberant lesions may explain, at least in part, the variable rates of positive biopsies observed with gastric cancers.Supported in part by a grant (AM07130) from the United States Public Health Service.     

Experimental toxicology of formaldehyde

Summary Formaldehyde is a reactive chemical which undergoes spontaneous reactions with various cellular constituents. Mutagenicity data may be interpreted on the background of this behavior. Mice are better able to reduce the irritating effect of formaldehyde than rats and to reduce their ventilation rate when formaldehyde acts on the respiratory tract. Subacute exposure of rats to concentrations higher than 2 ppm inhibits mucociliary clearance of the nasal epithelium and leads to progressive histological and ultrastructural lesions at this site. The occurrence of squamous cell carcinomas of the nasal epithelium of rats after 2 years inhalation of 14.3 ppm formaldehyde (CIIT study) is probably the result of chronic and recurrent local toxicity; this is supported by species differences in susceptibility to the tissue damaging and carcinogenic effect of formaldehyde (rat, mouse, hamster). Data on formaldehyde-DNA interaction further support the argument that a direct risk extrapolation from the formaldehyde effects in rats to those expected for man is not possible.Cancer Research and Clinical Oncology will publish in loose succession Editorials and Guest Editorials on topical and controversial problems in experimental and clinical oncology. These contributions will represent the personal opinion of the author. We would like to ask our readers to send their comments directly to the author.     

Unspezifische Phosphatasen und Esterasen im Melanoblastom der Chorioidea

Zusammenfassung Melanoblastome der Chorioidea bestehen aus wenig pigmentierten Melanocyten und stark pigmentierten als Makrophagen bezeichneten perivasculären Zellen. In sieben histochemisch untersuchten Melanoblastomen der Chorioidea besaßen die Makrophagen eine starke Aktivität unspezifischer Esterasen und saurer Phosphatasen, in den Melanocyten war die Aktivität schwach. Ein Melanoblastom vom epitheloiden Typ enthielt vorwiegend Zellen mit starker Enzymaktivität und zeigt außerdem alle Übergänge zwischen Melanocyten und Makrophagen. In allen Melanoblastomen waren die Fraktionen der isodynamen Esterasen und sauren Phosphatasen (Zymogramme nach Agarelektrophorese des Tumorextraktes) gleichartig, lediglich die Aktivität der einzelnen Fraktionen ist etwas verschieden. Die als Makrophagen bezeichneten Zellen sind ebenso wie die Melanocyten tumoreigene Zellen. Die starke Phosphataseaktivität wird auf den verstärkten Einbau von Phosphat (P³²) in die Tumorzellen bezogen.

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Summary The malignant melanoma of the choroid consists of melanocytes with slight pigmentation and of tumor cells (macrophage-like) that are heavily pigmented and perivascularly localized. In 7 malignant melanomas of the choroid belonging to the spindle cell and epithelioid type (Reese), the macrophage-like tumor cells histochemically showed a high activity of nonspecific esterases and acid phosphatases; the activity in melanocytes was low. A malignant melanoma of the epithelioid type chiefly contained tumor cells with high enzymatic activity and showed all transitional cells from melanocytes to macrophages. Zymograms of all melanomas (electrophoresis of tumor extract on agar-agar) showed similar fractions of isodynamic esterases and acid phosphatases which differed only in activity. The macrophage-like cells and the melanocytes were tumor cells. A relation presumably existed between the high activity of phosphatases and the increased incorporation of phosphate (P³²) into the tumor cells.

     

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     

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.     

Ultrastructural features of the plasma cells in “non-secretory” myeloma

Summary A case of non-secretory multiple myeloma is described. The diagnosis was based on the clinical picture, typical radiological findings, and infiltration of the bone marrow by myeloma cells which showed specific immuno-fluorescence staining mainly with antisera for IgM and kappa light chains. An attempt is made to explain the absence of pathological proteins in the serum, based on the ultrastructural findings of the myeloma cells, which showed buddings of the cell membranes containing endoplasmic reticulum and cytoplasmic material. It is suggested that the cells of the non-secretory type of multiple myeloma possess a normal excretory mechanism, but the pathological proteins are prevented to be secreted in the serum being surrounded by portions of the cell membrane.Established Investigator of the Chief Scientist's Office, Ministry of Health, Israel