Metabonomics of newborn screening dried blood spot samples: a novel approach in the screening and diagnostics of inborn errors of metabolism.

A novel, single stage high resolution mass spectrometry-based method is presented for the population level screening of inborn errors of metabolism. The approach proposed here extends traditional electrospray tandem mass spectrometry screening by introducing nanospray ionization and high resolution mass spectrometry, allowing the selective detection of more than 400 individual metabolic constituents of blood including acylcarnitines, amino acids, organic acids, fatty acids, carbohydrates, bile acids, and complex lipids. Dried blood spots were extracted using a methanolic solution of isotope labeled internal standards, and filtered extracts were electrosprayed using a fully automated chip-based nanospray ion source in both positive and negative ion mode.

[Neonatal screening for congenital hypothyreoidism in Germany. The development of concerned children in retrospect analysis using the federal state "Hessen"].

Background: Since widespread screenings for hypothyreosis were started in 1981 in Germany, the numbers of mental and physical handicaps due to hypothyroidism are reduced markedly. The aim of this study is to evaluate the actual efficiency of the newborn screenings in Germany using in the federal state "Hessen".

Methods/subjects: All children born between 1988 and 1992 with suspicions laboratory results in the screening examination were contacted personally and statements concerning the screening itself and the physical and mental development were gathered from their parents, doctors and teachers.

Neonatal screening for congenital hypothyroidism in Hessen, Germany: efficiency of the screening program and school achievement of 129 children at an age of 8-12 years.

Status and school achievement of 129 children born in Hessen between 1988 and 1992 and notified by a repeatedly elevated concentration of TSH in neonatal screening were evaluated. Interviews of mothers, teachers and pediatricians were used to score the development and educational achievements, respectively. A total of 298,175 newborns were screened and the incidence of permanent congenital hypothyroidism (PCH) was 1: 3,313 (n = 90).

Expression of fetal cytokeratins in epidermal cells and colloid bodies in lichen planus.

Clusters of immunoglobulin (Ig)-coated colloid bodies (CBs) in the dermo-epidermal zone are a typical immunohistochemical feature in lichen planus (LP)-lesions. They are considered to represent dyskeratotic basal keratinocytes, yet their composition has not been completely elucidated. In the present study, skin biopsies of 10 LP-lesions, 3 other dermatoses, and 10 biopsies of normal skin were studied immunohistochemically using monoclonal antibodies (MAbs) against fetal and differentiated epidermal antigens.

Effects of systemic complement activation on renal circulation of rats.

In a variety of immunopathological diseases activation of the complement cascade occurs either systemically or localized in the kidney. To elucidate the functional impact of complement activation upon the renal microcirculation, we administered cobra venom factor of Naja naja kaouthia (CVF) i.v.

Bioincompatibility--perspectives in 1993.

Bioincompatibility reactions related to the non-physiology of the procedure have plagued dialysis from its early days. Although the problem is certainly multifactorial, the present overview selectively focuses on some aspects of activation of late complement (C) components, the importance of which may have been underappreciated in the past. Dialysis patients are poised for intense C activation because of cumulation of the low molecular weight factor D, an intrinsically active serine esterase which is not inhibited by any known endogenous inhibitor and catalyzes an early step in the alternative pathway.

Complement deposits in epidermal cells after ultraviolet B exposure.

Ultraviolet B (UVB) radiation is known to induce formation of sunburn cells (SBC) in the epidermis. Since it was unknown whether this process might be accompanied by complement (C) activation, we analyzed C-deposition in skin biopsies taken before and 24 h and 48 h after UVB exposure (fourfold minimal erythema dose or fourfold minimal phototoxic dose) from 14 patients (5 receiving potentially photosensitizing drugs and 9 without such medication) by immunohistology. Local C-activation was visualized by direct or indirect immunofluorescence staining with polyclonal antibodies against C3b, C3d, C5, C9 and monoclonal antibodies to C3b, C3d, C9 and neoantigens on the terminal complement complex (TCC).

Effects of rC5a on the circulation of normal and split hydronephrotic rat kidneys.

Systemic release of complement-derived anaphylatoxin C5a is suggested to be involved in the pathogenesis of renal failure during endotoxic or severe traumatic shock. In the present study we analyzed renal hemodynamic effects of recombinant human complement 5a (rC5a) and examined whether these effects are mediated by the secondary release of other inflammatory mediators. Intravenous infusion of rC5a (0.

Expression of 1,25-dihydroxyvitamin D3 receptors in normal and psoriatic skin.

Increasing evidence suggests an immunoregulatory function of the potent steroid hormone 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) which has been successfully applied for treatment of psoriasis. The skin is both a site of production and a target of 1,25(OH)2D3. In vitro, 1,25(OH)2D3 inhibits proliferation and stimulates differentiation of keratinocytes.

Immunohistology of temporal arteritis: phenotyping of infiltrating cells and deposits of complement components.

Deposition of complement factors, immunoglobulins and infiltrating cells was evaluated by immunohistochemical staining in 30 temporal artery biopsy specimens from patients suffering from temporal arteritis and/or polymyalgia rheumatica and in controls. In the temporal arteritis group infiltrating cells, classic complement, alternative complement and lytic complex activation were detected. In specimens from patients suffering from only polymyalgia rheumatica there was unexpected evidence of classic complement and lytic complex activation.

Cysteinyl leukotriene actions on the microcirculation of the normal and split hydronephrotic rat kidney.

The effects of leukotriene D4 (LTD4) and leukotriene E4 (LTE4) on renal microcirculation were determined on normal and hydronephrotic female Wistar rats. In normal kidneys, the effects of LTD4 on total renal blood flow and glomerular filtration rate were measured by a flow meter and by inulin clearance. In the split hydronephrotic kidney, the LTD4- and LTE4-mediated vascular effects were localized by intravital microscopy.

[Hyalinosis cutis et mucosae (Urbach-Wiethe disease)--ultrastructural and immunological characteristics].

In this report on a young female patient with hyalinosis cutis et mucosae (Urbach-Wiethe disease, lipoid proteinosis), we present the clinical, immunological and ultrastructural features of this inherited disorder and discuss the differential diagnosis against other interstitial connective tissue depositions. Immunologically, the most important result was the increased amount of collagen type IV at the junction zones of epidermis, dermal vessels and appendages. This was in accordance with the ultrastructural deposition of hyalin material, mainly consisting of multiplied basal laminae at the respective junction zones.

Complement in inflammation: induction of nephritides and progress to chronicity.

The C5b-9 complex has a dual role as a factor involved in the initiation of nephritides and in the progress to chronicity and sclerosis. The unique pathophysiology of the membrane attack complex, distinct from other mediators, is its independence from specific receptors. It inserted in any membrane lipid bilayer tested so far.

Several epitopes on native human complement C9 are involved in interaction with the C5b-8 complex and other C9 molecules.

Ten monoclonal antibodies (mAb) against native human C9 exhibiting various inhibitory effects on the hemolytic activity of C9 (Bausback, J., Kontermann, R. and Rauterberg, E.

Fluid phase generation of terminal complement complex as a novel index of bioincompatibility.

Blood membrane interactions in hemodialysis have been shown to trigger complement (C) activation. As indicators of C-activation the anaphylatoxins (C3a and C5a) are problematical because of methodological difficulties and their kinetic properties. We developed a sensitive and specific micro-ELISA using a monoclonal antibody against neoantigens on the terminal complement complex (TCC); highly purified human TCC served as standard.

Assembly of terminal SC5b-9 complement complexes: a new index of blood-membrane interaction.

Activation of the complement system during the course of hemodialysis was recognized more than 20 years ago and since then the generation of C3a and C5a desarg has been used as parameters of blood-membrane interaction. More recently, determination of terminal C5b-9 complement complexes has become feasible. In the present study we determined plasma concentrations of C5b-9 complexes during hemodialysis using Cuprophan or Hemophan membranes.

Phototoxic erythema following PUVA treatment: independence of complement.

The effect of PUVA treatment on normal human serum (NHS), on isolated PMN, or on C3-deficient guinea pigs and congenic (C3-competent) control animals was tested. At a concentration of 0.1 or 1 mM/l 8-MOP and UVA doses of 5-30 J/cm2, PUVA failed to induce any detectable C3-cleavage in NHS.

Detection of human autoantibody against intercalated cells of kidney-collecting tubule.

Serum from a 24-year-old woman with a history of habitual abortions was examined for autoantibodies by indirect immunofluorescence microscopy. Fluorochrome-labelled antibodies to IgG revealed cytoplasmic staining of single cells in rat kidney collecting and connecting tubules. An identical staining pattern was reproducibly obtained in human and rabbit kidney, pointing to a cytoplasmic antigen concentrated in the apical pole of these cells.

N-deglycosylation of human complement component C9 reduces its hemolytic activity.

The effect of enzymatic deglycosylation of human complement component C9 on its hemolytic activity was investigated. Treatment of native C9 (Mr 71,000) with glyocpeptidase F (PNGase F) results in a stepwise decrease of the mol. wt.

Distribution of c-myc, c-myb, and Ki-67 antigens in interphase and mitotic human cells evidenced by immunofluorescence staining technique.

Using specific antibodies and the immunofluorescence staining technique we found a similar subcellular distribution pattern of the cellular proto-oncogene proteins c-myc and c-myb in interphase and mitotic HL60 and Molt4 cells. Antibodies against c-myc as well as those against c-myb protein gave rise to a nuclear staining excluding the nucleoli. In mitotic cells both proteins are apparently not associated with the chromatin of the condensed chromosomes, but appear diffusely distributed throughout the cytoplasm.