Identification and use of biomarkers in Gaucher disease and other lysosomal storage diseases

The value of biomarkers in the clinical management of lysosomal storage diseases is best illustrated by the present use of plasma chitotriosidase levels in the diagnosis and monitoring of Gaucher disease. The enzyme chitotriosidase is specifically produced and secreted by the pathological storage macrophages (Gaucher cells). Plasma chitotriosidase levels are elevated on average 1000-fold in symptomatic patients with Gaucher disease and reflect the body burden on storage cells. Changes in plasma chitotriosidase reflect changes in clinical symptoms. Monitoring of plasma chitotriosidase levels is nowadays commonly used in decision making regarding initiation and optimization of costly therapeutic interventions (enzyme replacement therapy or substrate reduction therapy). A novel substrate has been developed that further facilitates the measurement of chitotriosidase in plasma samples. Moreover, an alternative Gaucher-cell marker, CCL18, has been very recently identified and can also be employed to monitor the disease, particularly in those patients lacking chitotriosidase due to a genetic mutation. There is a need for comparable surrogate markers for other lysosomal storage diseases and the search for such molecules is an area of intense investigation.
Conclusion: The use of biomarkers can provide valuable insight into the molecular pathogenesis of LSDs, such as Gaucher disease and Fabry disease.     

The effect of adding group-based counselling to individual lifestyle counselling on changes in dietary intake. The Inter99 study – a randomized controlled trial

Background  

Few studies have investigated the specific effect of single intervention components in randomized controlled trials. The purpose was to investigate the effect of adding group-based diet and exercise counselling to individual life-style counselling on long-term changes in dietary habits.     

Antibodies to tumor necrosis factor alpha prevent increases in cell replication in liver due to the potent peroxisome proliferator, WY- 14,643

Several structurally dissimilar hypolipidemic drugs, plasticizers and halogenated hydrocarbons induce peroxisomes in hepatocytes, and cause hepatocellular adenoma and carcinoma in rats and mice. The mechanism by which these agents act is unknown, although recent studies have suggested a link between increased cell proliferation and hepatic cancer caused by peroxisome proliferators. Here, we demonstrate that neutralizing antibodies to tumor necrosis factor alpha (TNF alpha) block increases in protein kinase C and cell proliferation due to [4- chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid (WY-14,643), a hypolipidemic drug and potent peroxisome proliferator that causes tumors. WY-14,643 moderately elevated the level of TNF alpha mRNA in the liver. TNF alpha was detected immunohistochemically exclusively in Kupffer cells. These results demonstrate that WY-14,643 acts as an indirect mitogen on hepatocytes via TNF alpha. We propose that the Kupffer cell, a major source of TNF alpha in the liver, is involved in the mechanism of the mitogenic effect of WY-14,643.      

Evaluation of attempted prevention of unexpected infant death in very high-risk infants by planned health care

Three hundred and ninety-six babies born in Sheffield between 1982 and 1990 identified as being at "very high risk" of unexpected infant death by means of a scoring system, received an intensive programme of health care including a case discussion between a paediatrician, the GP and the health visitor held in the family doctor's surgery, weekly visits from the health visitor and informal hospital admission. Significantly fewer sudden unexpected infant deaths occurred in this group than were expected by logistic regression anlysis or occurred in the best available control group with comparable scores ( p = 0.024). Problems in evaluation include identification of an adequate control population, ethical difficulties in introducing a controlled study when the programme is already perceived as effective, and the calculation of "expected death rates". The results of this study indicate that very energetic programmes of intervention may prevent some deaths in vulnerable infants.     

Serum profiles of insulin-like growth factors and their binding proteins in adults with growth hormone receptor deficiency treated with insulin like growth factor I

Six adult patients with growth hormone receptor deficiency (GHRD) (2 men, 4 women) with an identical defect in the growth hormone receptor (GHR) gene, were treated with recombinant human insulin-like growth factor I (IGF-I), 40 μgikg S.C. twice daily, for 7 days. Serum concentrations of IGF peptide and IGF binding protein-3 (IGFBP-3) were measured by specific radioimmunoassays; serum IGFBPs were also measured by Western ligand blotting. The size distribution of both IGF-I and IGF-II was measured in serum following size-exclusion fast-performance liquid chromatography. IGF-I treatment resulted in a normalization of serum IGF-I levels on days 1–7 of treatment and a decrease in serum IGF-II levels. The fall in IGF-II levels and the simultaneous rise in IGF-I levels, however, resulted in an unchanged total serum IGF level. The low IGFBP-3 values did not significantly change during treatment, whereas there was a slight increase in IGFBP-2 levels. Preliminary analysis of size-fractionated sera suggested an increase in IGF-I levels in the 40 and 150 kDa regions at the expense of IGF-II levels. The results suggest that despite the failure of IGF-I treatment to increase IGFBPs significantly, serum IGFBP concentrations were sufficient to maintain normal levels of IGF-I. 0 Laron syndrome, growth hormone receptor deficiency, insulin-like growth factors, insulin-like growth factor binding protein     

Zur Enzymdiagnostik im Harn bei Kindern

Zusammenfassung Aus stationär aufgenommenen Kindern wurden 64 Patienten im Alter von 4 Tagen bis zu 13 Jahren randomisiert ausgewählt. Im Rahmen der Erstuntersuchungen wurden die Ausscheidung der Lactatdehydrogenase (LDH), alkalischen Phosphatase (AP) und der Leucinarylamidase (sogenannte Leucinaminopeptidase, LAP) im 8 Std-Nachtharn untersucht. Die Aktivitätsbestimmungen erfolgten in modifizierten Standardverfahren im Mikrolitersystem; der Sammlung, Vorbereitung und Aufarbeitung der Harne wurde besonderes Augenmerk geschenkt.Aus Patienten mit banalen, nichtentzündlichen Erkrankungen, ohne Fieber und ohne erhöhte Proteinausscheidung im Harn wurde ein Grundkollektiv von 18 Kindern gebildet. Bei diesem Kollektiv betrug die Ausscheidung, , für die LDH 2515±1838 mU/Std, für die AP 255±107 mU/8 Std und für die LAP 310±169 mU/8 Std. Mit zunehmendem Alter steigt die Ausscheidung der LDH von 1359 über 1531 und 2534 auf 4397 mU/8 Std an. Dagegen bleibt die Gesamtausscheidung der AP und LAP in diesem Alterszeitraum gleich. Bei allen Altersstufen ist die Ausscheidung der LDH dem Harnvolumen streng direkt proportional, r=0,95), nicht hingegen die der AP und LAP. Geschlechtsunterschiede sind bei allen drei Enzymen nicht festzustellen.Bei dem Krankenkollektiv bestehen auch bei unseren Kindern keine Zusammenhänge zwischen dem Bakteriengehalt und der proteinkonzentration und-ausscheidung einerseits und dem Ausmaß der Enzymurie andererseits. Dagegen wird Fieber über 38,6° C regelmäßig begleitet von hohen und höchsten Enzymausscheidungen bei AP und LAP; im Gegensatz dazu bleibt die LDH unauffällig. Umgekehrt bedeutet eine Enzymurie keinen Hinweis auf eine fieberhafte Erkrankung.Bei unseren Patienten mit Harnwegsinfekten und Nierenerkrankungen finden wir keine erhöhten Ausscheidungen der LDH, AP und LAP. Affallend ist die hohe Korrelation zwischen entzündlichen Allgemeinerkrankungen, insbesondere Entzündungen der Atemwege und des Magen-Darm-Traktes und der Ausscheidung der LAP, teilweise der AP. Zwischen Virusinfektionen und erhöhten Enzymausscheidungen scheinen keine Beziehungen zu bestehen.Die bisherigen Vorstellungen über die Mechanismen der pathologischen Enzymurie scheinen auf Grund des unterschiedlichen Verhaltens von LDH, AP und LAP einerseits, der verschiedenartigen Korrelation dieser Enzyme zu verschiedenen Krankheitstypen andererseits revisionsbedürftig. Weiterhin sollte gerade bei Kindern an Hand der vorgelegten Befunde der Versuch einer enzymatischen urinären Verlaufskontrolle von schweren Krankheitsbildern an größeren Fallzahlen durchgeführt werden. Ausgangspunkt hierfür sind die beobachteten normalen und schwerst pathologischen Enzymausscheidungen bei Pneumonien, Meningitiden und Leucosen.

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Diagnosis by means of enzymes in the urine of children

Sixty-four children aged from 4 days to 13 years were selected randomly amongst clinic patients. Parallel to primary examinations the urinary excretions of the enzymes lactate dehydrogenase (LDH), alkaline phosphatase (AP) and leucine aminopeptidase (LAP) during an eight-hour night period were measured. The determinations of activity were performed by modified standard methods in the microliter scale. Special attention was given to the collection and preparation of the urine samples. A group of 18 children with trivial diseases, with no signs of inflammatory processes, fever, or increased excretion of protein was formed. This group was used as a normal collective. The figures for the excretion of enzymes were as follows : 2515±1838 (LDH), 255±107 (AP) and 310±169 (LAP) mU/8 hrs. The excretion of LDH increased with advancing age, starting from 1359 to 1531, 2534 and 4397 mU/8 hrs. In contrast to these figures the total excretion of AP and LAP remains unchanged. At every age the excretion of LDH was closely correlated with the volume of the urine (coefficient of correlation r=0.95); there was no correlation with the excretion of AP and LAP. Nor did there seem to be any sex-dependent differentiation. In no case was any correlation found between the content of bacteria or proteins and that of enzymes in the urine. Elevated temperatures (38.6°C) were regularly accompanied by high values for the excretion of AP and LAP. Correlation between excretion of LAP, and, in some cases, of AP and general inflammatory diseases especially diseases of the respiratory and intestinal tracts was remarkably close. There did not appear to be any correlation will viral infections or — in our patients — to urogenital infections or kidney diseases. The hitherto genrally accepted conception of the mechanism of pathological excretion of enzymes seems to require a revision if one considers the different behaviour of LDH, AP and LAP.

     

Reaktionen des Nicotyrins mit 4-Dimethylaminobenzaldehyd

Reactions of Nicotyrine with 4-Dimethylaminobenzaldehyde The reactions resulting from nicotyrine with 4-dimethylaminobenzaldehyde in acidic solution were examined. At the molar ratio 2 + 1 the substances A and B and at the ratio 2 + 2 the substance B were formed. Substance A was recognized as bisnicotyrinyl-dimethylaminophenylmethane, substance B as a cyclohexadiene-1, 4-derivative of the structure mentioned in the text. The red-coloured compounds which arise when nicotyrine reacts with excess aldehyde could not be prepared in a pure form because of their light- and oxidation sensitivity. For the preparation of nicotyrine nicotine was reacted with sulphur. In contrast to the literature the product proved to be bisnicotyrinyldisulphide which could be reduced to the mercaptan. The disulphide on heating with powdered copper gave nicotyrine.