A screening for the occurrence of autoreactive antibodies in sera of pregnant and non-pregnant bitches.

Sera of healthy pregnant (group I, n = 11) and non-pregnant (group II, n = 11) bitches were screened for autoantibodies (AAb). In both groups, blood samples were drawn every fifth day between days 5 and 55 after mating. Serum was analysed via indirect immunofluorescence (IIF) with the Canine ANA HEp-2 Screening Kit. In all animals, anticytoplasmic AAb were detected. Utilizing primate-heart substrate slides AAb against contractile proteins of the cytoplasm could be observed. The predominating fluorescence pattern in pregnant animals resembled above all desmin, which was proven via Western blot. The sera were then pre-incubated with tropomyosin, actin, vimentin, vinculin and keratin solutions, and assessed on HEp-2 slides and on human and canine fibroblasts as well. The latter substrate was used to verify whether the detected Ab were in fact AAb. Utilizing tropomyosin, revealed elimination of the cytoplasmic fluorescences on all three substrates. It is therefore assumed, that in sera of healthy dogs, AAb against contractile structure proteins of the cytoplasm are present regularly. The majority of pregnant bitches presented with higher end-point titres (EPT), than to be found in non-pregnant dogs. AAb against desmin played the key role in those patterns. In addition, sera were screened for thyroid specific AAb, namely thyroglobulin, thyroid peroxidase (TPO), T3 and T4, and for AAb against insulin by ELISA or Western blot (TPO). Only in two of the pregnant bitches a weak positive reaction (1:100) for T3-AAb was detected.

Energy metabolism in graded perinatal asphyxia of the rat.

Although information on energy metabolism during hypoxemic-ischemic states is abundant, data on perinatal asphyxia (PA) are limited. As results from hypoxia-ischemia cannot be directly extrapolated to PA, a clinical entity characterized by acidosis, hypoxemia and hypercapnia, we decided to use a rat model of graded PA during delivery. Cesarean section was performed at the 21st day of gestation and the pups, still in the uterus horns, were asphyxiated from 0 to 20 minutes. In this model survival decreases with the length of asphyxia. Early changes of energy-rich phosphates in brain, heart and kidney were determined by HPLC. ATP and phosphocreatine gradually decreased with the length of asphyxia, with highest ATP depletion rate occurring in the kidney. ATP: brain 1.39 +/- 0.71 (0 min) to 0.06 microM/g wwt (20 min); heart 4.73 +/- 0.34 (0 min) to 1.08 +/- 0.47 (20 min); kidney 1.62 +/- 0.11 (0 min) to 0.02 +/- 0.02 (20 min). Phosphocreatine: brain 1.65 +/- 0.68 (0 min) to 0.51 +/- 0.45 microM/g (20 min); heart 6.98 +/- 0.38 (0 min) to 6.17 +/- 1.07 (20 min); kidney 8.23 +/- 0.86 (0 min) to 3.76 +/- 0.54 (20 min). We present data on energy derangement in a rat model of PA, closely resembling the clinical situation, showing that energy depletion precedes cell damage and death.

High levels of eosinophil cationic protein in wheezing infants predict the development of asthma.

BACKGROUND: In association with respiratory tract infections, infants may have episodes of wheezing, which represent the onset of asthma in some of them. Activated eosinophils play a central part in asthmatic inflammation. OBJECTIVE: We investigated whether, in infants experiencing their first episode of wheezing, eosinophil activation is present and can predict the development of asthma. METHODS: In a prospective trial, eosinophil activation was measured by eosinophil cationic protein (ECP) concentrations in serum from 33 nonatopic infants with their first episode of wheezing, 15 nonatopic infants with upper respiratory tract infection without wheezing, and 18 healthy nonatopic infants. One year later, the children were re-evaluated for a diagnosis of infantile asthma. RESULTS: Wheezing infants had higher median serum ECP levels (13.4 micrograms/L) than children with nonwheezy respiratory tract infection (7.6 micrograms/L, p < 0.005) or healthy subjects (7.1 micrograms/L, p < 0.005). In addition, wheezing infants (n = 13) with serum ECP concentrations greater than 20 micrograms/L were more likely to have asthma within 1 year than patients with ECP levels less than 20 micrograms/L (odds ratio = 12.4; confidence interval, 4.6-33.5). CONCLUSION: Eosinophil activation measured by serum ECP is present in infants with their first episode of wheezing illness, especially in those infants in whom asthma subsequently develops within 1 year. These data may indicate a predictive value of serum ECP measurements in children with wheezing to identify those patients in whom infantile asthma is developing. These findings probably also indicate that serum ECP may be used to identify the children who need early antiinflammatory treatment.

Reticulocyte maturity pattern analysis as a predictive marker of erythropoiesis in paediatrics. Part II: Pilot study for clinical application.

Reticulocyte quantification in peripheral blood samples is a commonly used diagnostic indicator of erythropoietic activity. A methodology based on flow cytometry additionally separates reticulocytes into 3 groups by fluorescence staining of the residual RNA. This identifies cells as 'high (HFR), medium (MFR) and low (LFR) fluorescence intensity' reticulocytes. In part II of the study we looked for the clinical applicability in paediatrics. Selected groups of patients with ineffective erythropoiesis, i.e. suffering from renal failure, oncologic patients with suppressed bone marrow activity caused by chemotherapy and anaemic new-born infants have been observed longitudinally for their reticulocyte maturity profiles. Data were compared to the commonly used parameters RBC, Hb and Hct. In all cases in which effective erythropoiesis returned documented by a normalization of standard blood parameters, HFR cells reacted significantly earlier than the traditional markers. These preliminary observations suggest the reticulocyte maturity pattern analysis can be used as an additional aid in diagnosis and as a helpful parameter for the monitoring of any anaemic situation.

Reticulocyte maturity pattern analysis as a predictive marker of erythropoiesis in paediatrics. Part I: Evaluation of age-dependent reference values.

Reticulocyte quantification in peripheral blood samples is a commonly used diagnostic indicator of erythropoietic activity. A methodology based on flow cytometry additionally separates reticulocytes into three groups by fluorescence staining of the residual RNA. This identifies cells as high (HFR), medium (MFR) and low (LFR) fluorescence intensity reticulocytes. In the present study an automated counter was evaluated and tested for its clinical applicability in paediatrics. In part I, reference intervals for different periods of childhood were determined. Except for the neonatal period there was no age-dependence so that children aged one week to 16 years have been summarized in one group. The wide variations we found in preterm children can be explained by different erythropoietic stimuli as the result of anaemia in infants with very low birthweight. No significant differences could be found between the sexes, not even at the onset of puberty. When using the reticulocyte maturity pattern analysis in clinical practice, the data give a helpful indication of the efficiency of the erythropoietic system.

Tonsillectomy and the immune system: a long-term follow up comparison between tonsillectomized and non-tonsillectomized children.

Immunological functions of the tonsils and possible effects of their removal are still controversial. One reason for this is the lack of long-term follow-up investigations after tonsillectomy. In the present study selected parameters of the cellular and humoral immune systems of 160 children 0.5-11 years after tonsillectomy (mean 6.6 +/- 2.1 years) were compared to those of 302 age-matched non-tonsillectomized children. In tonsillectomized children the incidence of infections of the upper respiratory tract was not increased compared to the non-tonsillectomized control group. Slightly increased percentages of CD 21 + cells, raised counts of CD4+ cells, absolute and relative increases in DR+ cells and a raised CD4+ DR count was found mainly in tonsillectomized boys, while lymphocyte subpopulations of tonsillectomized girls remained unaffected. Tonsillectomized children had lower IgA levels, but the complement system was not altered in either sex. These findings show that while tonsillectomy may lead to certain changes in the cellular and humoral immune systems, these alterations are clinically insignificant and no increased frequency of immunomodulated diseases should be expected.

Kappa/lambda ratio as parameter for evaluation of therapy-resistant epilepsy of childhood.

In a prospective study we tested the hypothesis whether an alteration in the immunoglobulin light chain kappa and lambda is an expression of an autoimmune mechanism. We investigated 152 children (84 boys, 68 girls, age 12 months to 16 years). Immunoglobulins were analyzed by kinetic nephelometry. The patients were grouped according to clinical and laboratory parameters. A high kappa/lambda ratio caused by high concentration of the kappa-chain was found in children with therapy-resistant epilepsy. 58% showed a high kappa/lambda ratio, in 28% no distinct association was possible and in 14% false-positive results were obtained. In seizure-free children immunological markers were specific in 71%, whilst 25% could not be characterized and 4% showed false-positive results. This alteration of immunoglobulins therefore can be used as a diagnostic parameter.

Pediatric and perinatal reference intervals for immunoglobulin light chains kappa and lambda.

In routine analysis for immunoglobulin light chains in pediatric diagnostics, the age-related reference intervals for serum kappa (kappa) and lambda (lambda) light chains were evaluated in 1543 healthy subjects (newborns to age 16 years, including 168 premature infants). Light-chain analysis was performed by rate nephelometry. IgG, IgA, and IgM were measured simultaneously, and heavy- and light-chain differences were calculated for control purposes. Results for IgG, IgA, and IgM generally agreed with reference intervals reported in the literature. kappa showed age-related changes comparable with changes in IgG concentrations, whereas lambda showed moderate fluctuations. The kappa/lambda ratio showed an almost linear increase with age, starting with 0.97 at four months and reaching the highest value of 2.21 at 15 years (mean values). Preterm infants presented with markedly low serum concentrations of IgG and corresponding light chains but with adult-type kappa/lambda ratios because of the maternal-origin IgG.

Human granulocyte elastase analysis on a clinical analyser (Greiner G-400) and comparison with the manual IMAC-Elastase method.

The performance of the human granulocyte elastase (EC 3.4.21.37) determination on the clinical analyser Greiner G-400 was evaluated. The detection limit of the assay is 4.68 micrograms/l. With pooled plasma samples, within-run variation (CV) ranged from 1.3 to 6.0%, between-run imprecision was 2.7 to 6.3%. Comparison with the homogenous immunoactivation manual assay using samples from 40 patients resulted in a very good correlation (r = 0.987). This suggests that the adapted instrumental method is able to replace the manual method for routine analyses.

[The mid-growth spurt--a pre-puberty growth spurt. Review of its significance and biological correlations]. / Der Mid-Growth-Spurt (MGS)--ein präpubertärer Wachstumsschub. Uberblick über seine Bedeutung und mögliche biologische Zusammenhänge.

The MGS is a growth phenomenon during early childhood, expressed by a mild transitory acceleration of growth velocity between five and eight years of age. It appears to be more pronounced in boys than in girls. It is probably caused by the functional maturation of the adrenals ("adrenarche") which leads to an increased androgen production during this age. Interactions with the pituitary growth hormone, also presenting with increased secretion rates at this particular period, are very probable. A hypothesis is offered for the explanation of individual differences and distinctness of the MGS. Although the MGS cannot be interpreted as a very first step of puberty, some additional biochemical facts suggest fundamental changes in the organism. Therefore, the MGS could be regarded as a "marker" within the biological development of the child.

Biological activity of alpha-alkyl-amino acids.

A series of alpha-alkyl-amino acids were tested for some biological functions in the mouse (OF-1 Himberg) by adding them to the animal chow (30 mg/kg/day) for a period of six weeks. No differences in fluid or food uptake could be observed during the feeding period, as compared to a control group. Histology of liver and kidney did not show any changes. Testing routine clinical chemical parameters (serum substrates and enzymes) revealed the following changes: Hex-Ala and But-Abu increased the serum glucose levels. But-Abu dramatically lowered the triglycerides. Serum albumin was increased by Pent-Ala, Me-Val, and But-Abu. LDH was inhibited by But-Abu.

Polymorphonuclear elastase in neonatal sepsis.

Quantitative analysis of polymorphonuclear elastase was carried out in 135 newborn infants during the first 28 days of life. In 15 neonates with septicaemia, significantly increased PMN-elastase activity (range 75 to 700 micrograms/l: P less than 0.0001) was observed at the time of recognition of infection. Cut-off level 65 micrograms/l. Normalisation of the elastase activities was observed only under real recovery of the patients. It is concluded that plasma elastase determination serves as a sensitive indicator of systemic infection in neonatology, and as a good parameter for monitoring the course of the disease, because PMN-elastase correlates with the clinical condition of the patient.