Modulation of chymase-mediated rat serosal mast cell degranulation by trypsin or diisopropyl fluorophosphate.

Exposure of rat serosal mast cells (RSMC) to chymase, an endogenous secretory granule serine protease, at 37 degrees results in exocytosis, as determined by beta-hexosaminidase release. As the number of RSMC is increased with a set amount of chymase, the net percentage beta-hexosaminidase release decreases linearly, implying a finite set of cellular interactions per chymase unit. Pretreatment of RSMC with trypsin at 37 degrees renders them refractory to subsequent exocytosis mediated by chymase in a dose- and time-dependent fashion, with complete refractiveness occurring by 15 min at 37 degrees with 2.5 micrograms trypsin/ml. Anti-IgE-mediated coupled activation-secretion of RSMC is not affected by the same trypsin pretreatment. When RSMC are pretreated with trypsin (2.5 micrograms/ml) for 0-120 min at 1 degree a progressive loss of sensitivity to activation by chymase at 37 degrees occurs. RSMC susceptibility to chymase-mediated degranulation after trypsin pretreatment can be partially regenerated by culturing the RSMC for about 24 hr in medium at 37 degrees. These findings suggest that a trypsin-sensitive constituent, possibly a receptor or substrate, is necessary for the functional interaction of chymase with RSMC. When added with diisopropyl fluorophosphate (DFP), chymase does not induce RSMC degranulation at 37 degrees. However, if the DFP is removed before addition of chymase at 37 degrees or is added after the chymase-priming event occurs at 1 degree, subsequent degranulation at 37 degrees is not inhibited. Thus, the induction and not the secretion phase is DFP-inhibitable in chymase-induced activation-secretion. In addition, the priming but not the exocytosis phase of chymase-initiated RSMC activation-secretion, which is not dependent on temperature and calcium ion concentration, involves a cellular trypsin-sensitive protein.     

Comparison of localized proton NMR signals of skeletal muscle and fat tissue in vivo: Two lipid compartments in muscle tissue

In vivo ¹H NMR spectra of small volumes-of-interest (VOI) were localized in human soleus muscle (8 ml) and compared with volume selective spectra of subcutaneous fat tissue and femoral yellow bone marrow (2 ml). All examinations were performed by the double spin echo (PRESS) localization technique. To provide comparability, spectra of different tissues were recorded using identical sequence timing. Clearly improved resolution of the lipid signals of muscle tissue was obtained using long echo times TE > 200 ms. The spectra of muscle tissue exhibit lipid signals that stem from two compartments with a difference of their resonance frequencies of about 0.2 ppm (Larmor frequency difference 12-13 Hz at 1.5 T). The existence of two fatty acid compartments is supported by measurements of the relaxation times and line shape analysis. Both compartments contain fatty acids or triglycerides with similar composition. Probably one compartment corresponds to fat cells within muscle tissue, the other compartment with lower Larmor frequency is located within muscle cells.     

Frakturen im h?heren Lebensalter: Wie sensitiv sind postalische Frageb?gen?

Zusammenfassung Im Rahmen der Europäischen Prospektiven Osteoporosestudie wurden zur Beurteilung der Validität postalischer Fragebögen zu Frakturen im höheren Lebensalter (50+) 144 Männer und Frauen Befragt, die in den vorangegangenen 12 Monaten in einem städtischen Krankenhaus wegen einer Fraktur behandelt worden waren. Die falsch negativen Antworten zur Frage nach einer Fraktur in diesem Zeitraum lagen mit 8% der Befragten niedriger als in anderen vergleichbaren Studien. Art und Häufigkeit der Befragung sind dabei von Bedeutung. Dagegen war das beigefügte Körperschema nur in zwei Drittel der Fälle geeignet, die Frakturlokalisation darzustellen. Eindimensionale Darstellung, geringe Grösse und fehlende Skeletteinzeichnung ergeben zu einem Drittel fehlerhafte Frakturlokalisationen.

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Fractures in the elderly: Are postal questionnaires sufficiently sensitive?

Summary Within the European Prospective Osteoporosis Study the validity of a postal questionnaire concerning fractures in the elderly was assessed. A sample of 144 men and women aged 50 to 84 hospitalized in an urban hospital due to fractures within the past 12 months was investigated. Eight percent of the respondents denied any recent fracture and turned out to be false negatives, less than previously recorded. Mode and frequency of questioning seem to influence the results. To assess fracture localisation, we used a graphical method (mannequin). Due to various factors, one third of all localisations were incorrect.

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Fractures des personnes âgrées: Quelle est la sensibilité des questionnaires envoyés par courrier?

Résumé Nous avons interrogé, dans le cadre de l'enquête européenne prospective sur l'ostéoporose, 144 hommes et femmes traités à la suite d'une fracture dans un hôpital communal dans les 12 mois précédents, afin d'évaluer les questionnaires envoyés par courrier et portant sur les fractures chez les personnes âgées (50 ans et plus). Les taux de faux négatifs dans les réponses à la question portant sur une fracture survenue dans cette période s'élevait à 8% des personnes interrogées, pourcentage plus bas que dans d'autres enquêtes analogues. Le type et la fréquence des interviews jouent un rôle important. Par contre l'adjonction d'une représentation schématique du corps humain ne permettait que dans deux tiers des cas de situer la fracture. Une représentation unidimensionelle, de taille réduite et l'absence de représentation du squelette sont à l'origine dans un tiers des cas des localisations incorrectes des fractures.

     

Health Impact Associated with Vertebral Deformities: Results from the European Vertebral Osteoporosis Study (EVOS)

To study the association between vertebral deformities and subjective health outcome indicators, including back pain and disability, a cross-sectional survey with spinal radiographs and personal interviews was carried out in 36 study centres in 19 European countries on a total of 15570 men and women aged 50–79 years (population-based stratified random samples). No interventions were done. The main outcome measures were the presence and intensity of current and previous back pain, functional capacity (ADL questionnaire) and overall subjective health. The presence and intensity of back pain and functional and health impairments varied within wide ranges with no obvious regional pattern. However, the associations between negative health outcomes and vertebral deformity were homogeneous between countries and between centres within countries. In logistic regression analyses weak but significant associations between the presence of vertebral deformities and various health indicators were demonstrated. The magnitude of the associations increased with severity and number of deformities. Compared with subjects without deformities those with low-grade deformities had no or only a weakly elevated risk for back pain, disability and impaired subjective health (odds ratios (OR) 1.2–1.3). The odds ratios increased for individuals with single severe deformities (OR 1.3–2.1) and were highest in those with multiple severe deformities (OR 1.7–4.2). The associations between vertebral deformities and negative health outcomes were stronger in men than in women. In this cross-sectional study radiologically assessed vertebral deformities were therefore weakly associated with both current and previous back pain as well as with functional and health impairments in both women and men. Multiple severe deformities were associated with severe and disabling back pain with stronger effects in men. Received: 27 December1997 / Accepted: 31 December 1997     

Survey response rates: national and regional differences in a European multicentre study of vertebral osteoporosis.

STUDY OBJECTIVE--This analysis aimed to compare the response rates of those invited to attend for screening in a multicentre, multinational study within Europe. DESIGN--This was a population survey. SETTING--Thirty four centres in 16 European countries. SUBJECTS--Men and women aged 50 years and over were recruited from population based sampling frames to participate in a prevalence survey of osteoporosis. Subjects were invited by post to attend for radiological screening and interview, and non-responders were followed up by repeat mailing. RESULTS--There was a substantial variation between centres in response rates: the mean was 49% and the range 5-83%. Adjusting for those known to have died or moved house did not affect the overall ranking. The response rates to each mailing also varied between centres: first mailing 45% (range 5-83%) and second mailing mean 10% (range 0-23%). The response rates varied in relation to age and sex and were higher in women than men. Rates fell gradually with age in women but rose in men until the age of 65 years. Response rates varied regionally. These were highest in countries from northern Europe and lowest in southern European countries, but there was wide variation both within regions and within countries. CONCLUSIONS--Multicentre, multinational studies within Europe will probably become increasingly popular. In this study, despite a standardised approach, the range in response rates between centres both within and between countries was substantial. Attempts at cross national standardisation in survey design can have only a limited effect on yielding uniformity in response.     

High Survivin Expression is Associated with Reduced Apoptosis in Rectal Cancer and May Predict Disease-Free Survival after Preoperative Radiochemotherapy and Surgical Resection

Background: Spontaneous apoptosis has been shown to predict tumor response to preoperative radiochemotherapy in rectal cancer. It remains to be elucidated, however, which genetic profile determines whether a tumor is more or less prone to apoptosis. Recently, a novel member of the inhibitor of apoptosis family, designated survivin, was identified. In the present study , we investigated the impact of survivin on tumor cell apoptosis and the risk to develop distant metastases or local failure after preoperative radiochemotherapy and surgical resection. Patients and Methods: The expression of survivin, p53, bcl-2 and the apoptotic index was evaluated by immunochistochemistry on pretreatment biopsies of 54 patients with locally advanced adenocarcinoma of the rectum. Survivin expression was correlated to clinical and histopathological markers, the levels of spontaneous apoptosis, p53 and bcl-2, as well as to disease-free survival, 5-year rates of local failure and distant disease after preoperative radiochemotherapy and surgical resection. Results: Survivin expression inversely correlated with the apoptotic index: High survivin expression was found in 56% of rectal carcinoma biopsies with a median apoptotic index of 1.22%. Conversely, low survivin expression in tumor cells was associated with a high median apoptotic index (2.29%, p=0.0001). High survivin expression also segregated with bcl-2 overexpression (65% bcl-2+ in tumors with high survivin expression as compared to 35% bcl-2+ in tumors with low survivin expression), but the difference was not statistically significant (p=0.1). Low survivin expression was significantly related to an increased disease-free survival rate (77% vs 18% at 5 years in tumors with high survivin expression, p=0.02) and to a redued risk for distant metastases (18% vs 78% at 5 years in tumors with high survivin expression, p=0.05) and local failure (6% vs 37% at 5 years in tumors with high survivin expression, p=0.07). Conclusion: An inverse correlation between survivin expression and the level of spontaneous apoptosis in pretreatment biopsies suggests that survivin strongly inhibits tumor cell apoptosis in rectal cancer. Survivin expression may provide a novel predictive indicator for disease-free survival after preoperative radiochemotherapy and surgical resection in rectal cancer. Hintergrund: Für die spontane Apoptoserate konnte eine prädiktive Bedeutung für die Tumorantwort nach präoperativer Radiochemotherapie beim Rektumkarzinom gezeigt werden. Bisher ist nicht geklärt, welches genetische Profil die Apoptosefähigkeit eines Tumors bestimmt. Kürzlich wurde das Protein Survivin als ein neues Mitglied der Inhibitor-of-Apoptosis-Familie identifiziert. In der vorliegenden Studie wurd der Einfluss von Survivin auf die Apoptoserate sowie das Fernmetastasenrisiko und die Rate lokoregionärer Rezidive nach neoadjuvanter Radiochemotherapie und Operation des Rektumkarzinoms untersucht. Patienten und Methoden: Die Expression von Survivin, p53, bcl-2 und die Apoptsoerate wurden immunhistochemisch in prätherapeutischen Biopsien von 54 Patienten mit lokal fortgeschrittenen Adenokarzinomen des Rektums bestimmt, die einheitich nach einem neoadjuvanten Radiochemotherapieprotokoll behandelt wurden. Die Survivin-Expression wurde mit klinischen und histopathologischen Markern, dem p53- und bcl-2-Index, dem Apoptoseindex, dem krankheitsfreien Überleben sowie dem Auftreten von Fernmetastasen oder lokoregionären Rezidiven korreliert. Ergebnisse: Die Survivin-Expression korrelierte invers mit dem Apoptoseindex: Bei hoher Survivin-Expression (56% aller Tumoren) betrug der mediane Apoptoseindex 1,22%, bei niedriger Survivin-Expression 2,29% (p=0,0001). Eine erhöhte Survivin-Expression war auch mit einer bcl-2-Überexpression assoziiert (65% bcl-2+ in Tumoren mit hoher Survivin-Expression, 35% bcl-2+ in Tumoren mit niedriger Survivin-Expression, p=0,1). Bei niedriger Survivin-Expression betrug das krankheitsfreie Überleben nach 5 Jahren 77%, bei hoher Survivin-Expression 18% (p=0,02). Die Fernmetastasenrate und die Rate an lokoregionären Rezidiven betrugen nach 5 Jahren bei Tumoren mit niedriger bzw. hoher Survivin-Expression: 18% vs. 78 % (p=0,05) bzw. 6% vs. 37% (p=0,07). Schlussfolgerung: Die signifikant inverse Korrelation zwischen der Survivin-Expression und der spontanen Apoptoserate in prätherapeutischen Biopsien weist darauf hin, dass Survivin beim Rektumkarzinom die Apoptose der Tumorzellen inhibiert. Die Survivin-Expression kann als neuer prädiktiver Indikator für das krankheitsfreie Überleben nach präoperativer Radiochemotherapie und Operation gelten.     

A genotype-controlled analysis of plasma dopamine beta-hydroxylase in healthy and alcoholic subjects: evidence for alcohol-related differences in noradrenergic function.

BACKGROUND: Norepinephrine and dopamine mediate important aspects of alcoholism and alcohol withdrawal. Dopamine-beta-hydroxylase (DbetaH) converts dopamine to norepinephrine. A recent study demonstrated a strong association between variance in plasma DbetaH activity and a novel polymorphism (DBH-1021C-->T) at the structural locus (DBH) encoding DbetaH protein. METHODS: Our study investigated whether the DBH-1021C-->T polymorphism and plasma DbetaH activity were associated with alcoholism or with delirium tremens (DT) during alcohol withdrawal by analyzing 207 German alcoholic and 102 healthy control subjects. We also examined the influence of the polymorphism on enzyme activity. RESULTS: Mean (+SD) plasma DbetaH activity measured in alcoholic subjects abstinent was significantly lower than that observed in control (27.7 + 16.7 vs. 35.6 + 18.8; p =.01). It did not differ between subjects with DT during withdrawal and subjects with mild withdrawal symptoms. The T allele of the DBH-1021C-->T polymorphism was significantly associated with lower plasma DbetaH activity. None of the alleles or genotypes were associated with alcoholism or DT. CONCLUSIONS: The data indicate that the alcoholism-related reduction in plasma DbetaH activity is independent of genotype at DBH-1021C-->T and replicate the finding that DBH-1021C-->T is strongly associated with plasma DbetaH activity in a native Western European population.     

Whom to treat? The contribution of vertebral X-rays to risk-based algorithms for fracture prediction. Results from the European Prospective Osteoporosis Study

Introduction Vertebral fracture is a strong risk factor for future spine and hip fractures; yet recent data suggest that only 5–20% of subjects with a spine fracture are identified in primary care. We aimed to develop easily applicable algorithms predicting a high risk of future spine fracture in men and women over 50 years of age.Methods Data was analysed from 5,561 men and women aged 50+ years participating in the European Prospective Osteoporosis Study (EPOS). Lateral thoracic and lumbar spine radiographs were taken at baseline and at an average of 3.8 years later. These were evaluated by an experienced radiologist. The risk of a new (incident) vertebral fracture was modelled as a function of age, number of prevalent vertebral fractures, height loss, sex and other fracture history reported by the subject, including limb fractures occurring between X-rays. Receiver Operating Characteristic (ROC) curves were used to compare the predictive ability of models.Results In a negative binomial regression model without baseline X-ray data, the risk of incident vertebral fracture significantly increased with age [RR 1.74, 95% CI (1.44, 2.10) per decade], height loss [1.08 (1.04, 1.12) per cm decrease], female sex [1.48 (1.05, 2.09)], and recalled fracture history; [1.65 (1.15, 2.38) to 3.03 (1.66, 5.54)] according to fracture site. Baseline radiological assessment of prevalent vertebral fracture significantly improved the areas subtended by ROC curves from 0.71 (0.67, 0.74) to 0.74 (0.70, 0.77) P=0.013 for predicting 1+ incident fracture; and from 0.74 (0.67, 0.81) to 0.83 (0.76, 0.90) P=0.001 for 2+ incident fractures. Age, sex and height loss remained independently predictive. The relative risk of a new vertebral fracture increased with the number of prevalent vertebral fractures present from 3.08 (2.10, 4.52) for 1 fracture to 9.36 (5.72, 15.32) for 3+. At a specificity of 90%, the model including X-ray data improved the sensitivity for predicting 2+ and 1+ incident fractures by 6 and 4 fold respectively compared with random guessing. At 75% specificity the improvements were 3.2 and 2.4 fold respectively. With the modelling restricted to the subjects who had BMD measurements (n=2,409), the AUC for predicting 1+ vs. 0 incident vertebral fractures improved from 0.72 (0.66, 0.79) to 0.76 (0.71, 0.82) upon adding femoral neck BMD (P=0.010).Conclusion We conclude that for those with existing vertebral fractures, an accurately read spine X-ray will form a central component in future algorithms for targeting treatment, especially to the most vulnerable. The sensitivity of this approach to identifying vertebral fracture cases requiring anti-osteoporosis treatment, even when X-rays are ordered highly selectively, exceeds by a large margin the current standard of practice as recorded anywhere in the world.This work was presented in part at the 30th European Symposium on Calcified Tissues, 8–12 May 2003, Rome, Italy.A.J. Silman and J. Reeve are the EU Grant holders and Project Leaders.     

EEG-results in early-treated children with phenylketonuria 4 through 10 years under treatment (author's transl)]

EEG-results in 21 children with Phenylketonuria (PKU), put on a diet at least since their 3rd month of life, now aged 4 through 10 years, with normal psychomotor development, lacking abnormal neurological signs, are compared with the results in 796 healthy children of the same age. Visual evaluation and frequency analysis of the background activity of the EEGs reveal no differences between children with PKU and controls. Generally, there is no close correlation between mean plasma levels of phenylalanine (phe-means) during treatment, and the composition of the EEG when combined age groups are compared. But a trend can be demonstrated: In the small group of 8 years old children phe-means up to 6 mg-% can be associated with a faster (alpha), and phe-means above 6 mg-% with a slower (theta) background activity. The frequency of metabolic derailment (single phe-values above 10 mg-%) does not correlate with the EEG. Focal and generalized hypersynchronous activity (HSA) is observed significantly more often even in early treated, normally developing children with PKU. In children aged 4--8 years, HSA is associated with a high proportion of slow waves compared with those who do not display HSA. In children 9 and 10 years old, however, no such differences can be seen. It is hypothesised that even early treated children with PKU, in their first years show a retardation in the development of their background activity but catch up by 9--10 years. It still remains uncertain whether further age-appropiate development of the EEG can be observed after liberating or discontinuing the dietary regimen at this age.