Holoprosencephaly and low molecular weight proteinuria: the human homologue of murine megalin deficiency.

We encountered a child with holoprosencephaly, pulmonary insufficiency, absent circulating vitamin D metabolites, mild albuminuria, and urinary excretion of vitamin D-binding protein. The child displayed a phenotype highly reminiscent of that observed in mice genetically deficient for megalin, a member of the low-density lipoprotein receptor superfamily. Only the Guthrie card was available from the child; the DNA sufficed for a limited haplotype analysis. We were not able to implicate the megalin gene locus directly; however, the possibility of a functional megalin defect in this child remains. To the best of our knowledge, this patient represents the first report that pathologic abnormalities consistent with megalin deficiency are present in humans.

Short stature in a mother and daughter with terminal deletion of Xp22.3.

Short stature in females is often caused by hemizygosity for the terminal portion of Xp due to monosomy X or a deletion. We report on a mother and daughter with short stature as sole phenotypic abnormality and deletion of bands Xp22.32-p22.33 demonstrated by classic and molecular cytogenetic analysis. In both individuals, the deleted X chromosome was late replicating. Molecular analysis suggested that the deletion is terminal and the breakpoint was localized between the STS and DXS7470 loci in Xp22.32. Chromosome analysis is often done on females with short stature to exclude Ullrich-Turner syndrome. Small deletions, terminal or interstitial, are easily missed by conventional cytogenetic investigation; thus molecular analyses are useful to detect those cases.

Kidney involvement in a 17-year-old boy with eosinophilic fasciitis.

Eosinophilic fasciitis (EF) is characterized by symmetrical scleroderma-like induration of skin over one or more distal extremities, peripheral eosinophilia, absence of Raynaud phenomenon and visceral involvement and a favourable response to systemically administered corticosteroids. Like other scleroderma-like disorders EF is rarely described in children. We report renal involvement in a 17-year-old boy with EF. Urinalysis disclosed proteinuria. Prior to corticosteroid therapy renal biopsy was performed which revealed ischemic collapse of glomerular capillaries and atrophy of tubules of the cortex. Electron-microscopic studies showed hyperplasia of the renin-producing epitheloid cells in the juxtaglomerular apparatus. Few other publications have depicted renal involvement in EF of quite different character. In these cases renal biopsy and histological classification is warranted because of prognostic and therapeutic implications.

[The synergistic effect of delta-aminolevulinic acid and photodynamic laser therapy based on an in vitro model of the ATP tumor chemosensitivity test]. / Synergistischer Effekt von Delta-Aminolävulinsäure und photodynamischer Lasertherapie anhand eines In-vitro-Modells mit dem ATP-Tumorchemosensitivitätstest.

Photodynamic laser therapy (PDT) is under prospective clinical evaluation in gynecology. We used the ATP tumorchemosensitivity assay to detect the efficacy of delta-aminolevulinic acid (ALA) in combination with PDT. Two invasive cervical cancer cell lines were used, C33 and SIHA. The cell-damaging effect was quantified by measuring the intracellular ATP content. The 50% inhibitory concentration of ALA was 760 microM in the cell line C33 and 500 microM in the cell line SIHA. The combination of ALA incubation and PDT was synergistic in both cell lines. After in vitro experiments with this method, the combination is now used in a clinical phase II study. PDT after topical ALA application promises to become a possible alternative for the treatment of preinvasive and invasive malignancies in gynecology.

[Use of photodynamic laser therapy in gynecology]. / Einsatz der photodynamischen Lasertherapie in der Gynäkologie.

OBJECTIVE: Dysplasia of the vulva and uterine cervix are often multicentric, tend to recur and have mostly to be treated by surgical procedures. The photodynamic laser therapy (PDT) may be an alternative, which selectively destroys neoplastic tissue after topical delta-aminolevulinic acid (ALA) used as a photosensitizer. METHODS: We measured the distribution of fluorescence in dysplastic and nondysplastic tissue after topical application of ALA in 27 patients with dysplasia. In 3 vulvar and 2 cervical dysplasias, PDT was performed. RESULTS: We could show a selective enrichment of endogenous porphyrins in dysplastic tissues, whereas benign tissue showed no fluorescence. The fluorescence was limited to the mucosa. The heterogeneous fluorescence pattern was influenced by the duration of ALA application. In the treated patients, cytological and clinical parameters showed improvement after use of PDT. The longest recurrence-free interval from treatment up to date is 15 months (range 3-15 months). CONCLUSIONS: After first results of penetration studies and clinical follow-up, PDT after topical ALA application seems to be a good alternative to surgical procedures in dysplastic changes of the genital tract.

Non-surgical management of a ruptured posttraumatic pancreatic pseudocyst in a child.

Generally speaking, isolated pancreatic injuries are rare after abdominal blunt trauma. However, the incidence of pancreatic injuries in children has risen in recent decades. Pancreatic pseudocyst represents a typical complication after acute pancreatitis due to blunt abdominal trauma. Spontaneous rupture of pseudocysts leading to acute abdominal pain has been described, however, it rarely occurs, especially in pediatric patients. We report the successful non-surgical management of a ruptured pancreatic pseudocyst in a 5-year-old girl which occurred 27 days after trauma. The traumatic acute pancreatitis was due to a handlebar injury.

[Rhythmic behavior of a colorless mutant of Euglena gracilis]. / Das rhythmische Verhalten einer farblosen Mutante von Euglena gracilis.

A colorless mutant of Euglena gracilis shows a cireadian rhythmic mobility in darkness just as mixotrophic and autotrophic green forms do.Therefore the obligatory heterotrophic form is very suitable for testing to what extent the rhythmic behavior of the mixotrophic cells, in contrast to that of the autotrophic cells, is independent of photosynthesis.Like the green cells, the colorless cells are synchronized by a single transient from light to dark. About 12 hours after the beginning of the darkness the rhythmic mobility of the colorless cells attains the first maximum, whereas the green forms both show their first maximum 18 hours after the end of the light period, which is much more advantageous for a photosynthesizing organism.The free running period seems to be dependent on the temperature during anaerobic glycolysis and independent of temperature during respiration, just as it has been recently found out in the case of green forms. The type of energy supply changes with the age of the cultures.Respiration has no significance as energy source for the rhythm which continues under pure glycolytic conditions.A sudden increase of the constant temperature does not shift the phase. The same is true in the case of mixotrophic cells but not in the case of autotrophic cells. However, the rhythm is often first suppressed for several days.A lowering of the temperature is followed by two or three transients with about half the frequency, but it does not influence the phase as far as it can be extrapolated. The same has been shown to be true in the case of mixotrophic cells.The rhythmic behavior of the heterotrophic cells is very similar to that of the mixotrophic ones. However, the heterotrophic cells are very sensitive to changes in temperature, which are more compensated for in the mixotrophic cells, apparently by photosynthesis.

[Hemorrhagic shock and encephalopathy syndrome in infancy and childhood]. / Hämorrhagisches Schock-und Enzephalopathiesyndrom im Säuglings- und Kindesalter.

After a few days of prodromal illness with diarrhea and fever there was an abrupt onset of coma, seizures, severe shock, and hyperpyrexia in two infants; they than developed bleeding, pancytopenia, and impairment of liver and renal function. Both infants survived with major neurologic sequelae. The clinical symptoms and laboratory parameters were compatible with hemorrhagic shock and encephalopathy syndrome which was first described in 1983. The clinical course and biochemical features of this disease and its differential diagnoses are described, and etiology and pathogenesis are discussed. Furthermore it will be discussed, whether the hemorrhagic shock and encephalopathy syndrome is actually a "new" disease or just a new label for a known disorder.

Proteinuria after renal transplantation: diagnosis with highly sensitive silver stain in sodium dodecylsulphate-polyacrylamide gradient gel electrophoresis.

Sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of urinary proteins is becoming increasingly significant in monitoring renal allograft recipients. After conventional Coomassie Blue staining, changes in renal proteinuric patterns during acute renal transplant rejection could not be seen in many cases. Discrimination between rejection and nephrotoxic side effects of cyclosporine overdosage was also not possible. We therefore examined the possible advantages of a highly sensitive silver staining technique in this study. A total of 734 urine samples obtained from 38 patients after allogenic kidney transplantation were examined by SDS-PAGE with consecutive Coomassie Blue and silver staining. Twenty-two histologically proven rejections and 20 cyclosporine overdosage episodes were diagnosed in these patients within a time period of up to 524 days after transplantation. No changes in proteinuric patterns were seen in 9 of 22 patients after Coomassie stain during rejection, and only 12 cases showed a rise of glomerular protein bands, whereas silver stain revealed an increase in 19 of 22 cases. Discrimination between cyclosporine overdosage and rejection was possible with a probability of p less than 0.001 after silver stain when using the changes in the number of glomerular protein bands as a criterion. These findings suggest that application of a highly sensitive silver stain instead of the conventional Coomassie stain after SDS-PAGE reflects considerable progress in monitoring renal allograft recipients.

Fetal serum concentrations of cystatin C and beta2-microglobulin as predictors of postnatal kidney function.

OBJECTIVES: Cystatin C and beta(2)-microglobulin are established serum markers of renal function in children and adults. In contrast to creatinine, diaplacental exchange is minimal. The aim of the study was to establish reference values in fetal serum and to test their efficiency in predicting postnatal kidney function. STUDY DESIGN: This was a prospective noninterventional study measuring cystatin C and beta(2)-microglobulin by particle-enhanced immunoturbidimetry in excess serum from 129 cordocenteses performed in 84 fetuses. Reference intervals (mean +/- 1.96 SD) were calculated in a subgroup of 54 fetuses without evidence of kidney disease, and these reference values were evaluated in 75 sera from 55 fetuses. RESULTS: Mean cystatin C was 1.66 +/- 0.202 mg/L (upper limit 2.06), and mean beta(2)-microglobulin was 4.25 +/- 0.734 mg/L. Unlike cystatin C, beta(2)-microglobulin decreased significantly with gestational age so that the upper reference limit was 7.19-0.052 x gestational age in weeks. beta(2)-Microglobulin had higher sensitivity (90.0% vs 63.6%) and cystatin C a higher specificity (91.8% vs. 85.5%) for the prediction of impaired renal function; diagnostic efficiency was equal (87.6% vs. 86.1%). Fetuses with impaired renal function at birth or who were aborted for renal malformations had higher cystatin C concentrations than those in a control group. beta(2)-Microglobulin was increased only in fetuses who were aborted. CONCLUSION: Fetal serum cystatin C and beta(2)-microglobulin concentrations may be useful predictors of postnatal kidney function.

Evaluation of pediatric nephropathies by a computerized Urine Protein Expert System (UPES).

A computerized Urine Protein Expert System (UPES) measuring creatinine, total protein, albumin, IgG, alpha(1)-microglobulin, alpha(2)-macroglobulin, and N-acetyl-beta-D-glucosaminidase, together with urine dipstick testing for granulocyte esterase and hemoglobin pseudoperoxidase, and measurement of serum creatinine had been found to be useful in adults for differentiating between renal disorders. The objective of this study was to investigate UPES for identifying the different types of proteinuria and their underlying prerenal, glomerular, tubular, and postrenal causes in 146 children characterized by routine and invasive nephrological investigations. UPES proved to be a useful tool in pediatric renal patients after refinements were implemented in the program. Comparing UPES with the pediatric nephrologist's interpretation of all available clinical and laboratory data, UPES diagnosed glomerulopathies in 46 (75%) of 61 patients. In a further 23% it suggested glomerular involvement by indicating either a disturbed glomerular permeability or increased excretion of albumin. Tubular proteinuria was correctly described by UPES in 23 (100%) patients with different tubulopathies. UPES revealed normal kidney function in all healthy children and all children with remission of renal disorders. Therefore, UPES can be regarded as a useful tool in the automated differentiation of renal diseases in children.

Oscillometric twenty-four-hour ambulatory blood pressure values in healthy children and adolescents: a multicenter trial including 1141 subjects.

Ambulatory blood pressure (ABP) monitoring is increasingly used to evaluate the blood pressure of children and adolescents. The upper normal ABP values in the pediatric age group are still unknown, because reference values based on a sufficiently high number of healthy children have not yet been published. In this multicenter trial, we pooled ABP records of 1141 healthy children and adolescents with a body height between 115 and 185 cm. The study was carried out by seven centers according to a common protocol. The 50th percentile for 24-hour systolic ABP increased moderately with height, from 103 to 113 mm Hg in girls and from 105 to 120 mm Hg in boys. The 50th percentile for diastolic 24-hour means was 66 +/- 1 mm Hg, irrespective of height or gender. Diastolic daytime means were 73 +/- 1 mm Hg, which is remarkably high compared with reference values for casual blood pressure. The mean nocturnal systolic and diastolic ABP (midnight to 6 AM) was 13% +/- 6% and 23% +/- 9% lower compared with the daytime means (8 AM to 8 PM), respectively. This multicenter study provides well-based limits of normal ABP in mid-European children.

Quality of salivary tears following autologous submandibular gland transplantation for severe dry eye.

BACKGROUND: This study aimed to characterise the composition of the pre-ocular fluid after transplantation of the autologous submandibular gland (SMG) for patients with severe dry eye. METHODS: Stimulated and unstimulated pre-ocular fluid from 15 patients (17 eyes) with a viable SMG graft ("SMG-salivary tears"), as well as normal tears and SMG saliva (20 normal subjects/ 20 eyes), was sampled. As global tear parameters, fern pattern analysis and SDS gel electrophoresis were performed. As specific quality parameters, total protein content, secretory immunoglobulin A (SIgA), lysozyme, amylase, sodium, potassium and osmolality were measured using routine laboratory methods. The flow rate of SMG-salivary tears was determined in 5 patients by means of sequential scintillography. RESULTS: The fern pattern of SMG-salivary tears was coarse and thus more similar to normal SMG saliva than tears. SDS gel electrophoresis of the SMG-salivary tears showed albumin and two unidentified proteins in addition to the normal tear pattern. Osmolality and total protein content of SMG-salivary tears were higher than in normal SMG saliva, but still lower than in normal tears. High activities of normal tear antibacterial proteins (SIgA, lysozyme and amylase) were detected in the salivary tears. Stimulation of the secretion did not alter the composition of SMG-salivary tears. The flow rate of SMG-salivary tears was closer to that of normal tears than normal SMG saliva. CONCLUSION: Salivary tears resulting from SMG-transplantation represent condensed SMG saliva. Thus their quality is intermediate between normal tears and normal SMG saliva. High levels of secretory proteins demonstrate that the gland maintains an active function. Surgical denervation and residual tear components from the ocular surface are the most likely factors to cause the complex differences between normal SMG saliva and SMG-salivary tears. The effects of this secretion on the ocular surface are currently being evaluated in a clinical and laboratory study.

Pharmacokinetics and selectivity of aminolevulinic acid-induced porphyrin synthesis in patients with cervical intra-epithelial neoplasia.

Photodynamic therapy (PDT), due to its tumor selectivity, represents an alternative approach to diagnose and treat cervical intra-epithelial neoplasia (CIN) without altering normal surrounding tissue. Our aim was to investigate the pharmacokinetics and the selectivity of 5-aminolevulinic acid (5-ALA)-induced porphyrin fluorescence after topical administration, to obtain basic clinical data for future diagnostic fluorescence imaging and PDT protocols for CIN. Twenty-eight non-pregnant women with a cytological diagnosis of low-grade or high-grade squamous intra-epithelial lesions were included. An aqueous solution containing 3% 5-ALA was topically applied 1 to 6 hrs prior to conization using a cervical cap. After excision, porphyrin-induced fluorescence was quantified in dysplastic (n = 14) and normal epithelium (n = 28) by means of quantitative fluorescence microscopy. High values of porphyrin fluorescence were found in squamous epithelium between 150 and 450 min, with a maximum at 300 min following administration of 5-ALA. Ratios of porphyrin fluorescence of dysplastic vs. surrounding normal epithelium were 1.3 and 1.21 for CIN 1 (n = 3) and CIN 2 (n = 3), respectively. In CIN 3 patients (n = 8), this ratio was 2.35; the best selectivity of 5-ALA-induced porphyrin fluorescence in CIN 3 lesions (ratio 3) was observed with a topical administration time of between 150 and 250 min. Our results demonstrate that patients with CIN 3 show higher 5-ALA-induced fluorescence compared with normal epithelium. The optimal administration time of topically applied 5-ALA was between 3 and 4 hr. Our data suggest that topical ALA-PDT and photodynamic diagnosis might be suitable for detecting CIN.

Quality of salivary tears following autologous submandibular gland transplantation for severe dry eye.

BACKGROUND: This study aimed to characterise the composition of the pre-ocular fluid after transplantation of the autologous submandibular gland (SMG) for patients with severe dry eye. METHODS: Stimulated and unstimulated pre-ocular fluid from 15 patients (17 eyes) with a viable SMG graft ("SMG-salivary tears"), as well as normal tears and SMG saliva (20 normal subjects/20 eyes), was sampled. As global tear parameters, fern pattern analysis and SDS gel electrophoresis were performed. As specific quality parameters, total protein content, secretory immunoglobulin A (SIgA), lysozyme, amylase, sodium, potassium and osmolality were measured using routine laboratory methods. The flow rate of SMG-salivary tears was determined in 5 patients by means of sequential scintillography. RESULTS: The fern pattern of SMG-salivary tears was coarse and thus more similar to normal SMG saliva than tears. SDS gel electrophoresis of the SMG-salivary tears showed albumin and two unidentified proteins in addition to the normal tear pattern. Osmolality and total protein content of SMG-salivary tears were higher than in normal SMG saliva, but still lower than in normal tears. High activities of normal tear antibacterial proteins (SIgA, lysozyme and amylase) were detected in the salivary tears. Stimulation of the secretion did not alter the composition of SMG-salivary tears. The flow rate of SMG-salivary tears was closer to that of normal tears than normal SMG saliva. CONCLUSION: Salivary tears resulting from SMG-transplantation represent condensed SMG saliva. Thus their quality is intermediate between normal tears and normal SMG saliva. High levels of secretory proteins demonstrate that the gland maintains an active function. Surgical denervation and residual tear components from the ocular surface are the most likely factors to cause the complex differences between normal SMG saliva and SMG-salivary tears. The effects of this secretion on the ocular surface are currently being evaluated in a clinical and laboratory study.