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In this study we show that mitochondrial uncoupling protein 1 (UCP1) in brown adipose tissue (BAT) and thymus mitochondria can be ubiquitinylated and degraded by the cytosolic proteasome. Using a ubiquitin conjugating system, we show that UCP1 can be ubiquitinylated in vitro. We demonstrate that UCP1 is ubiquitinylated in vivo using isolated mitochondria from brown adipose tissue, thymus and whole brown adipocytes. Using an in vitro ubiquitin conjugating–proteasome degradation system, we show that the cytosolic proteasome can degrade UCP1 at a rate commensurate with the half-life of UCP1 (i.e. 30–72 h in brown adipocytes and ~ 3 h, in thymocytes). In addition, we demonstrate that the cytoplasmic proteasome is required for UCP1 degradation from mitochondria that the process is inhibited by the proteasome inhibitor MG132 and that dissipation of the mitochondrial membrane potential inhibits degradation of UCP1. There also appears to be a greater amount of ubiquitinylated UCP1 associated with BAT mitochondria from cold-acclimated animals. We have also identified (using immunoprecipitation coupled with mass spectrometry) ubiquitinylated proteins with molecular masses greater than 32 kDa, as being UCP1. We conclude that there is a role for ubiquitinylation and the cytosolic proteasome in turnover of mitochondrial UCP1. This article is part of a Special Issue entitled: 17th European Bioenergetics Conference (EBEC 2012).

This paper suggests that there is probably a dominant, sex-linked type of chondrodysplasia punctata. Clinical data are reported for three girls with such a disorder. Two of their mothers showed a mild form of cicatricial alopecia. The pathognomonic dermatological findings in the children include erythematous skin changes and striated ichthyosiform hyperkeratosis during the first months of life. Later on, patterned ichthyosis, follicular atrophoderma, coarse, lusterless hair and cicatricial alopecia become evident. It is assumed that about one fourth of all cases with chondrodysplasia punctata reported in the literature belong to the dominant sex-linked type.

Previous researchers have demonstrated that 3,4-methylenedioxymethamphetamine (MDMA) induced hyperthermia, in skeletal muscle of animals, is uncoupling protein 3 (UCP3) dependent. In light of our investigations that in vivo phosphorylation of UCP1 is augmented under conditions of cold-acclimation, we set out to investigate whether (a) UCP3 was phosphorylated in vivo and (b) whether in vivo phosphorylation of UCP3 resulted in increased proton leak following MDMA administration to animals. Our data demonstrate that MDMA treatment (but not PBS treatment) of animals results in both in vivo serine and tyrosine phosphorylation of UCP3 in skeletal muscle mitochondria, isolated in the presence of phosphatase inhibitors to preserve in vivo phosphorylation. In addition, proton leak is only increased in skeletal muscle mitochondria isolated from MDMA treated animals (in the presence of phosphatase inhibitors) and the increased proton leak is due to phosphorylated UCP3. UCP3 abundance in skeletal muscle mitochondria is unaffected by MDMA administration. Preservation of UCP3 phosphorylation and increased proton leak is lost when skeletal muscle mitochondria are isolated in the absence of phosphatase inhibitors. We conclude that MDMA treatment of animals increases proton leak in skeletal muscle mitochondria by activating UCP3 through in vivo covalent modification of UCP3 by phosphorylation. Furthermore, we deduce that the MDMA induced hyperthermia in skeletal muscle is due to increased proton leak in vivo as a result of activation of UCP3 through phosphorylation.

Using the Elisa-Test of Dakopatts, Hamburg, described by Ishiguro et al and modified by us (Mikrotitration plates instead of tubes, blocking up free bonding capacities in the plates with 1% gel fluid, altered incubation periods) we determined secretory IgA (SIgA) in saliva samples of 376 infants and children. The probands could be divided in three groups: Group 1, serving as controls, consisted of 163 healthy children. Group 2 comprised 111 children suffering from acute infection of the respiratory tract. Group 3 consisted of 102 children with chronic airways diseases, in particular, asthma. In the healthy infants and children we found age dependent increases of SIgA until the age of 4 years. The median values amounted 16.7 (newborns), 59.2 (1st year), 118.2 (2nd year), 149.2 (3rd year), 185.5 (4th year), 159 (5th year) and 175.8 mg/1 (5th-13th year). A similar age dependent increase of SIgA was evident in the saliva samples of children suffering from acute infections of the respiratory tract. In the children with chronic airways diseases there was only a slight increase of SIgA during the first 4 years (x = 78.0-113.5 mg/I) and an abrupt (statistically significant) rise in the fifth year. The median value of SIgA was 216 mg/I in the children aged 5-13 years. Serum IgA along with salivary IgA were measured in 128 children (r = 0.40, p

This review of the literature presents the pooled data from several perinatal centers throughout the world published during the last three years. It regards the longterm prognosis of extremely low birthweight infants (less than 1000 g). Since the introduction of neonatal intensive care the mortality rate dropped from greater than 90% to less than 50%. On the other hand the morbidity rate improved only slowly. Recent reports from the Anglo-American literature show that among the survivors 13.7% are seriously and 25.1% mildly handicapped. 60-70% of the extremely low birthweight infants, despite of normal IQ's, are experiencing school problems. Most of them have cognitive, perceptual and motor deficits combined with behaviour problems ("the new morbidities"). The incidence of cerebral palsy remained unchanged (6-8%) over the last two decades. Until now, there is no significant effect of the surfactant therapy on the morbidity rate. The rate of serious longterm disability increases with decreasing birthweight. The results of the Anglo-American and German follow-up studies differ only slightly.

A case of a 10-year old boy suffering from asthma bronchiale following pleuropneumonia is reported. Paradoxically, bronchospasmolysis tests using physiological saline + salbutamol or ipratropium bromide impaired the lung function of this patient. Salt solutions inhaled in increasing doses until 1.4% generated severe bronchoconstriction, however, inhalation of DNCG + salbutamol normalized the lung function completely.

Using the Elisa-Test of Dakopatts, Hamburg, described by Ishiguro et al and modified by us (Mikrotitration plates instead of tubes, blocking up free bonding capacities in the plates with 1% gel fluid, altered incubation periods) we determined secretory IgA (SIgA) in saliva samples of 376 infants and children. The probands could be divided in three groups: Group 1, serving as controls, consisted of 163 healthy children. Group 2 comprised 111 children suffering from acute infection of the respiratory tract. Group 3 consisted of 102 children with chronic airways diseases, in particular, asthma. In the healthy infants and children we found age dependent increases of SIgA until the age of 4 years. The median values amounted 16.7 (newborns), 59.2 (1st year), 118.2 (2nd year), 149.2 (3rd year), 185.5 (4th year), 159 (5th year) and 175.8 mg/l (5th-13th year). A similar age dependent increase of SIgA was evident in the saliva samples of children suffering from acute infections of the respiratory tract. In the children with chronic airways diseases there was only a slight increase of SIgA during the first 4 years (mean = 78.0-113.5 mg/l) and an abrupt (statistically significant) rise in the fifth year. The median value of SIgA was 216 mg/l in the children aged 5-13 years. Serum IgA along with salivary IgA were measured in 128 children (r = 0.40, p less than 0.001). 6 children had a complete IgA deficiency and 4 children an incomplete IgA deficiency, i.e. low secretory IgA levels in saliva (36.8-50.0 mg/l) and lacking IgA in serum (less than 14 mg/dl).(ABSTRACT TRUNCATED AT 250 WORDS)

The protective effects of inhaled MgS04 on exercise-induced asthma were studied in 13 children aged 6-13 years. The patients performed 6 min. running test on 3 separate days. Spirometry (FEV1, SRAW, VK, PF) was recorded on each test day 10 min. after medicament inhalation directly before and instantly, 2, 5, 10 and 20 min after the running test. t-tests on the mean revealed no significant effects on spirometric measurements (p less than 0.1), but in 6 patients MgS04-inhalation prevented FEV1--decreases greater than 20%. The combination gave better protection compared to salbutamol alone demonstrated from the measurements after exercise testing. Thus, MgS04-inhalation could be useful in the protective treatment of asthma.

Oxypurine analysis was done in the CSF of 190 children with different diseases. The patients could be divided into four groups: Group A, serving as controls, consisted of 56 children suffering from diseases without neurological signs, for example, leukaemia. 16% of them had raised hypoxanthine values greater than 7.5 mumol/l and 32% raised uric acid values greater than 12.0 mumol/l. Group B comprised 97 children suffering from diseases with neurological signs, for example, meningitis. For these patients the frequency of raised hypoxanthine and uric acid values in the CSF was twice as high as in Group A. Group C comprised 31 patients with different forms of cerebral convulsions. Among these patients 52% had raised hypoxanthine and 70% raised uric acid values. The findings of these patients are described in a previous paper (Manzke et al. 1981). Group D comprises 6 patients from whom CSF samples were taken postmortally. All these deceased patients showed extremely high hypoxanthine + xanthine and uric acid concentrations in their CSF.

Serum creatinine, uric acid, and hypoxanthine and xanthine concentrations were determined in 17 mother-infant pairs at the time of delivery. Creatinine and uric acid levels were nearly similar, but hypoxanthine and xanthine were more than twice as high in the blood of the infants than in the blood of their mothers. In the same newborns the urinary excretion of creatinine, uric acid, hypoxanthine and xanthine, uracil, cAMP, and cGMP was measured on the first and fourth day of life. Creatinine, uracil, and cAMP increased, hypoxanthine and xanthine, and cGMP decreased, whereas the output of uric acid was nearly the same on both days. Correlations of the excreted substances to each other were calculated.

Severe anaphylactic reactions were observed in 2 children, aged 2 and 14 years, with acute lymphocytic leukemia and a 12 month old infant with a functional T-cell defect after intravenous administration of a polyethyleneglycol (PEG)-treated immunoglobulin preparation. All 3 children suffered additionally from hypogammaglobulinemia. Intravenous infusions of 7S-immunoglobulin preparations with a modification of the Fc portion, however, had been well tolerated by the same patients. Investigations of the immunoglobulin preparations revealed IgG aggregates (approximately 35%) in one batch of the PEG-treated immunoglobulin preparation. Prekallikrein activator and kallikrein were not increased in the immunoglobulin preparations tested. These results in addition to our clinical experiences suggest that only 7S-immunoglobulin preparations with a modification of the Fc portion should be used in patients with hypo- or a-gammaglobulinemia, in order to avoid complement activation released by the intact 7S-preparations.

Two healthy volunteers were treated with hypoxanthine 3 x 1 g and allopurinol 3 x 100 mg daily for 1 week. During this treatment serum oxypurine concentration and urinary oxypurine excretion increased as expected. No side effects were observed except for some mild daytime drowsiness and lethargy. Measurements of urinary serotonin (5-HT) excretion showed decreases to as much as 60% below initial values. Decreased urinary 5-HT excretion was also found in a patient with incomplete Lesch-Nyhan syndrome during treatment with high doses of hypoxanthine. His neurological symptoms improved slightly. The results suggest that high doses of hypoxanthine exert a nonspecific sedative effect on both patients with Lesch-Nyhan syndrome and healthy controls. The cause is probably a reduced synthesis or release of 5-HT.

Es wird ein in Anlehnung an die Methode von Gleichmann et al. modifiziertes Mikroverfahren zur Bestimmung des intracellularen pH in Erythrocyten beschrieben. Bestimmungen des intracellularen Bicarbonats sind mit dieser Methode mit nur geringer Fehlerbreite ebenfalls moglich. Das Mesverfahren basiert auf dem Astrup-Prinzip unter Verwendung des Nomogramms von Siggaard-Andersen.

Es wird uber eine atypische Pneumonie bei drei Fruhgeborenen berichtet, die mit langanhaltenden lympho-monocytaren Blutbildveranderungen einherging. Im Liquor fand sich eine Pleocytose. Die Atiologie blieb unklar. Trotz ausgedehnter bakteriologischer und virologischer Untersuchungen lies sich kein einheitlicher Erreger nachweisen.

Im Capillarblut von Fruhgeborenen wurden Sauerstoffdruck, Kohlensaurespannung, pH, Standardbicarbonat und Basenuberschus bestimmt. Die pH-Mittelwerte von 78 Kindern bei der Aufnahmeuntersuchung ergaben, aufgeteilt in drei Gruppen, Fruhgeborene ohne Atemnotsyndrom pH 7,35±0,055, uberlebende Fruhgeborene mit Atemnotsyndrom pH 7,22±0,09, verstorbene Fruhgeborene mit Atemnotsyndrom pH 7,19±0,05. Der Unterschied zwischen der ersten und zweiten Gruppe ist hochsignifikant, zwischen der zweiten und dritten Gruppe nicht signifikant. Eine statistisch gesicherte Korrelation zwischen dem Ausmas der Acidose und dem Grad der Untertemperatur bei unterkuhlten Fruhgeborenen lies sich nicht errechnen (n=114, r=+0,11).

Die durch hohe Dosen gonadotroper Hormone hervorgerufenen primaren ovariellen Veranderungen und die dadurch bedingte spatere Umgestaltung der Gonaden wurden beim Menschen und bei verschiedenen Tieren einer eingehenden histologischen Untersuchung unterzogen.

The cerebral damage of infants born in breech presentation can be classified by cause into three major groups.

Cardiazol induced seizures in rabbits showed that the highest oxypurine concentrations can be detected in the CSF 1 hour after the convulsions. There is a sharp decline continuing until the third hour. After that the CSF values remain nearly constant until the 24th hour being about ten times higher than in the controls. There is a good correlation of these results obtained through the densitometric thin-layer, enzymatic-oxymetric, and HPLC-methods. Creatinine and potassium were raised only during the first two hours postconvulsively. Uracil appeared in the CSF slightly higher at the 1 hour and at the 12 and 24 hour values. A parallel increase of the oxypurine and creatinine concentrations was found in the serum between 30 to 120 minutes postconvulsively. After that the raised serum values decreased slowly to the initial values. CSF samples were examined in 31 children postconvulsively: Hypoxanthine was found to be raised in 8 of 12 patients with severe grand mal seizures, 1 of 6 patients with hypsarrhythmia, 1 of 8 patients with short seizures (less than 2 min) and in all 5 patients with petit mal status. In contrast to these groups the hypoxanthine concentrations was raised only in 2 of 20 children with aseptic meningitis. The difference between the group of children with convulsions and the group of children with aseptic meningitis is significant (p less than 0.005). Also, the frequency of raised uric acid concentration is higher in the group of children with convulsions (70%) than in the group of children with aseptic meningitis (40%); (p less than 0.05).

Among 1115 newborns who died during 1957--1976, 136 (12.2%) showed macroscopic renal uric acid infarctions. The incidence depended on the age of the infants and their fluid supply during the first days of life. After introduction of parenteral alkali-glucose infusion in the treatment of perinatal complications, the incidence of renal uric acid infarctions decreased from 19.3% to less than 1.0%. Analysis of the oxypurines in the renal uric acid infarctions of three newborns revealed high percentages of hypoxanthine (31.1%, 21.8% and 11.0%) along with the uric acid. Hypoxanthine ratios above 15% retrospectively point to chronic hypoxia of the newborn.

Within the sample of 1783 children from the prospective study "Schwangerschaftsverlauf und Kindesentwicklung" (Course of pregnancy and development of children) followed-up until the age of 6 years the mothers of 182 suffered from late gestosis (10.2%). The values of the systolic blood pressure and of the protein concentration in the urine of the mothers measured at their last examination before onset of labour were crucial for the diagnosis of gestosis. According to this definition 95 mothers with systolic pressure greater than or equal to 150 mm Hg were classified as hypertensive (group of the hypertension children) and 87 mothers with both systolic pressure greater than or equal to 150 mm Hg and proteinuria greater than 1 g/l as being preeclamptic (group of the preeclamptic children). The control groups contained the same number of children (matched pairs). There were no significant differences in the findings between the different groups. Apparently, the prognosis of the child's development is favourable provided that the gestosis does not last long and does not lead to placental insufficiency.

Severe anaphylactic reactions were observed in 2 children, aged 2 and 14 years, with acute lymphocytic leukemia and a 12 month old infant with a functional T-cell defect after intravenous administration of a polyethyleneglycol (PEG)-treated immunoglobulin preparation. All 3 children suffered additionally from hypogammaglobulinemia. Intravenous infusions of 7S-immunoglobulin preparations with a modification of the Fc portion, however, had been well tolerated by the same patients. Investigations of the immunoglobulin preparations revealed IgG aggregates (approximately 35%) in one batch of the PEG-treated immunoglobulin preparation. Prekallikrein activator and kallikrein were not increased in the immunoglobulin preparations tested. These results in addition to our clinical experiences suggest that only 7S-immunoglobulin preparations with a modification of the Fc portion should be used in patients with hypo- or a-gammaglobulinemia, in order to avoid complement activation released by the intact 7S-preparations.

Report of an 1 1/2 year old boy with the typical features of Williams-Campbell syndrome: chest deformity (thorax piriformis), pulmonary hyperinflation, ballooning of the segmental and subsegmental bronchi during inspiration and collapse with expiration. Atelectasis of the left upper lobe after bronchiolitis obliterans. The literature reports 23 patients with Williams-Campbell syndrome. There is every reason to believe that the extensive bronchiectasis are not due primarily to developmental deficiency of the bronchial cartilage, as the most authors assume, but result from destructive changes after inflammation. In our case very likely an adenovirus infection was the cause. The clinical features of the syndrome look like a severe therapy resistant asthma bronchiale.

This description of the physiology and pathophysiology of the calcium and phosphate balance in neonates is based on our own studies and a short review of the literature. In general, the higher the calcium concentration in the umbilical cord blood the greater its decrease during the first two days of life. With asphyxiated newborns the decrease in the serum level of both the total calcium and the ionized calcium surpasses that in nonasphyxiated newborns by approximately a third. There are various causes for neonatal hypocalcemia. The most striking causes for the early form of hypocalcemia are likely to be a transient hypoparathyroidism or a failure of end-organ responsiveness. In nearly all newborns we found a low urinary cAMP excretion on the first day of life increasing significantly until the fourth day. Measurements of the urinary cAMP excretion are an appropriate parameter for recognizing the parathormone effect. By measuring both the parathormone level in the serum and the urinary cAMP excretion it is possible to distinguish hypoparathyroidism from pseudoparathyroidism . Several cases with different forms of hypocalcemia are discussed.

The concentration of hypoxanthine, xanthine and uric acid in the first 24-h urine of 105 newborn infants was measured densitometrically by thin-layer chromatography. 45 of them had moderate or severe perinatal complications. Among these newborns, 26 infants with perinatal complications (58 %) and 4 infants without perinatal complications (7 %) had an elevated urinary excretion rate of hypoxanthine. Urinary xanthine was not increased, uric acid was slightly higher in the group of infants with perinatal complications. It seems that a crucial mark is involved, if the rate of hypoxanthine exceeds 15% of the total urinary oxypurine excretion, or, if related to urinary creatinine, more than 0.075 μmol hypoxanthine/μmol creatinine. Apparently, with hypoxic newborns increased values of urinary hypoxanthine excretion can be used to quantify the lack of oxygen retrospectively.

Oxypurine analysis was done in the CSF of 190 children with different diseases. The patients could be divided into four groups: Group A, serving as controls, consisted of 56 children suffering from diseases without neurological signs, for example, leukaemia. 16% of them had raised hypoxanthine values greater than 7.5 mumol/l and 32% raised uric acid values greater than 12.0 mumol/l. Group B comprised 97 children suffering from diseases with neurological signs, for example, meningitis. For these patients the frequency of raised hypoxanthine and uric acid values in the CSF was twice as high as in Group A. Group C comprised 31 patients with different forms of cerebral convulsions. Among these patients 52% had raised hypoxanthine and 70% raised uric acid values. The findings of these patients are described in a previous paper (Manzke et al. 1981). Group D comprises 6 patients from whom CSF samples were taken postmortally. All these deceased patients showed extremely high hypoxanthine + xanthine and uric acid concentrations in their CSF.

The metabolism of newborn infants during their first week of life is characterized by an increased breakdown of nucleoproteins leading to transient hyperuricemia and hyperuricosuria which is particularly enhanced in infants with hemolytic diseases. Furthermore, there are physiologically considerable changes in the concentration of calcium in the blood of newborns.

In 55 children with congenital heart defects (aged 6 months and older), hematocrit, hemoglobin concentration, acid-base status, oxygen pressure, oxygen saturation, and 2,3-DPG content of erythrocytes were determined in blood taken from the vena cava superior. The data were correlated with each other as well as with parameters obtained during cardiac catheterization. Whereas an increased hematocrit was seen only at a difference in arteriovenous oxygen saturation of more than 30%, or a central venous saturation below 65%, a close correlation between 2,3-DPG concentration in erythrocytes and these parameters was obtained already in the normal range (r = 0.79, and r = 0.78, respectively). A relatively close correlation was also found between 2,3-DPG concentration and half-maximal oxygen saturation pressure (T50 value; r equals 0.73), and the T50 values and the central venous oxygen saturation (r equals 0.61), respectively. Because of the shift to the right of the O2-binding curve the functional oxygen transport capacity was increased by an average 15% in children with acyanotic heart defects (mean T50 value of 31.6 mm Hg), and by 40% in children with cyanotic heart defects (mean T50 value of 34.6 mm Hg), as compared to healthy children (mean T50 value of 28.1 mm Hg). A marked influence on the position of the O2-binding curve and hence on oxygen release of the blood in the periphery is therefore exerted by the 2,3-DPG concentration, whose effect was about twice that of the Bohr effect in the pH range measured.

Nevus flammeus (teleangiectaticus) is the most common variety of congenital nevus. Among 3961 infants checked at the ages of 3--7 days and again at 6 weeks 42% showed nevi flammei mediales in the nape and/or in the eyelids and glabella. The mothers of the infants presenting with n.fl. tended to be overweight, to have circulatory distrubances and prolonged duration of laor. The infants with n.fl. frequently had concomittant edema, navel hernia and minor degenerative stigmata. However malformations did not occur more commonly. This investigation suggests that exogenous factors such as diseases during pregnancy, intake of drugs and difficulties in delivery do not influence the genesis of n.fl. Only constitutional factors could be correlated with the incidence of n.fl. Since there was a higher frequency in female than in male infants and a lower frequency in premature than in term newborns it may be that hormonal inffluences increase the incidence of n.fl., too. These results were obtained from the prospective study: Schwanger-schaftsverlauf und Kindesentwickling (DFG).

Extremely low birth weight infants are particularly prone to rickets (osteopenia) due to their rapid growth and to deficient intake of calcium and phosphate. In some premature infants suffering from phosphate depletion hypercalcemia syndrome may precede bone demineralisation. Additionally, the adverse effects of calciprivic drugs (phenytoin, phenobarbital, glucocorticoids, furosemide, heparin) contributing to the development of neonatal rickets are discussed. Phosphorus depleted or heparin treated experimental animals develop impairment of mineralisation as manifested by rickets or osteomalacia. Some clinical cases of neonatal rickets are reported and a dosage schedule for parenteral infusion of minerals is given.