Hepatic syndecan-1 changes associate with dyslipidemia after renal transplantation.

Syndecan-1 is a transmembrane heparan sulfate (HS) proteoglycan present on hepatocytes and involved in uptake of triglyceride-rich lipoproteins via its HS polysaccharide side chains. We hypothesized that altered hepatic syndecan-1 metabolism could be involved in dyslipidemia related to renal transplantation. In a rat renal transplantation model elevated plasma triglycerides were associated with fivefold increased expression of hepatic syndecan-1 mRNA (p < 0.01), but not protein. Expression of syndecan-1 sheddases (ADAM17, MMP9) and heparanase was significantly up-regulated after renal transplantation (all p < 0.05). Profiling of HS side chains revealed loss of hepatic HS upon renal transplantation accompanied by significant decreased functional capacity for VLDL binding (p = 0.02). In a human renal transplantation cohort (n = 510), plasma levels of shed syndecan-1 were measured. Multivariate analysis showed plasma syndecan-1 to be independently associated with triglycerides (p < 0.0001) and inversely with HDL cholesterol (p < 0.0001). Last, we show a physical association of syndecan-1 to HDL from renal transplant recipients (RTRs), but not to HDL from healthy controls. Our data suggest that after renal transplantation loss of hepatic HS together with increased syndecan-1 shedding hampers lipoprotein binding and uptake by the liver contributing to dyslipidemia. Our data open perspectives toward improvement of lipid profiles by targeted inhibition of syndecan-1 catabolism in renal transplantation.

Effect of amelogenin ECM protein on the healing of chronic leg ulcers with atrophie blanche.

OBJECTIVE: To determine the effect of topically applied amelogenin extracellular matrix protein(AEMP) in patients with non-healing venous leg ulcers combined with atrophie blanche. METHOD: This retrospective case series of patients with non-healing venous leg ulcers with atrophie blanche of the distal proportion of their lower legs, where non-healing was defined as no progress toward healing for 3 months previously, under standard therapy. Patient records were reviewed for associated diseases, wound diagnoses, distal blood pressure, previous treatments and changes in wound area. Patients were treated with AEMP once a week, for a period of 12 weeks, or until full healing. RESULTS: Eleven patient records were reviewed retrospectively. The median age of the patients was 81 years (range 40-95 years), with a mean wound size of 4.7 ± 3.Scm2 and median wound duration of 6 months (range 3-444 months).AII patients had venous or combined arterial/venous insufficiency. After 12 weeks' treatment with AEMP, complete healing, defined as I 00% re-epithelialisation, was documented in four patients (36%), marked improvement(> SO% epithelialisation) in three patients (54%, 55% and 83% wound closure, respectively), slight improvement in one patient (9.4% wound closure), no change for two patients and worsening in one.AEMP was well tolerated, and no patients reported side effects. CONCLUSION: The results of this retrospective study suggest that AEMP improves healing in chronic venous leg ulcers combined with atrophie blanche.

Virilization of the external genitalia and severe mental retardation in a girl with an unbalanced translocation 1;18.

A female infant with dysmorphic facial features, psychomotor retardation, and clitoris hypertrophy is described. Molecular cytogenetic analyses revealed a de novo unbalanced translocation, causing partial monosomy 1p36 and partial trisomy 18q22. Monosomy 1p was confirmed by FISH, and trisomy of the distal part of chromosome 18q was demonstrated by microFISH. Gene copy number changes in these chromosomal regions were determined by array-CGH. The absence of a number of facial dysmorphic signs, and the presence of clitoris hypertrophy indicate that the combination of a del(1p36->pter) with a dup(18q22->qter) may lead to a unique phenotypic constellation. The findings at birth and at age 12 years in our patient are compared with genotype-phenotype correlations discussed in the literature.

Antagonism of HSV-tk transfection and ganciclovir treatment on chemotherapeutic drug sensitivity.

Our study focused on the influence of herpes simplex virus thymidine kinase (HSV-tk) expression and ganciclovir (GCV) treatment on the sensitivity of C6 glioma cells to frequently used chemotherapeutic drugs, i.e. adriamycin (ADR), cisplatin (CDDP), 5-fluorouracil (5-FU), and methotrexate (MTX). Transfection with HSV-tk revealed an increased sensitivity to GCV and CDDP and a decreased sensitivity to ADR and MTX. No significant differences were found in sensitivity to 5-FU. Combined treatment in a HSV-tk negative cell line revealed an additive effect when GCV was combined with ADR, whereas an antagonistic effect was found when GCV was combined with CDDP, 5-FU, or MTX. Comparable results were obtained in an HSV-tk positive cell line, apart from CDDP, which showed an additive effect. In conclusion, both HSV-tk transfection and subsequent GCV treatment can influence the sensitivity of tumor cells to various chemotherapeutic drugs in an antagonistic manner. Therefore, combining HSV-tk/GCV gene therapy with chemotherapy might not always be beneficial.

Prothrombin time ratio is reduced by magnesium contamination in evacuated blood collection tubes.

Magnesium ions were detected in sodium citrate solutions in several lots of evacuated blood collection tubes. The mean concentrations ranged between 1.3 and 1.6 mmol/L. Magnesium was also present in the rubber stoppers of the blood collection tubes and could be leached into the citrate solution. It was shown that magnesium added to citrated plasma shortened the prothrombin time of both coumarin and normal plasma. The effect of magnesium was relatively greater on coumarin than on normal plasma resulting in reduced prothrombin time ratio. Shortening of the prothrombin time was also observed when magnesium chloride was added to dialysed plasma, i.e., in the absence of citrate. These results indicate that magnesium contamination can interfere with accurate INR determination in the control of oral anticoagulant therapy.

Latent X-ray damage in the rat sciatic nerve results in delay in functional recovery after a heat treatment.

The influence of X-irradiation on the sensitivity of the rat sciatic nerve to local hyperthermia was investigated. A 10 or 20 mm long segment of the nerve was irradiated intraoperatively using 50 kV X-rays. Hyperthermia (30 min at 45 degrees C), was applied to the irradiated part (over a length of 5 mm) of the nerve using a brass thermode. Functional damage to the nerve was assessed using the toe-spreading test, which mainly assesses the motor function of the sciatic nerve. Radiation alone (doses up to 70 Gy) did not lead to detectable damage for at least 90 weeks. Hyperthermia alone (30 min at 45 degrees C) resulted in complete loss of motor function. This function loss was transient and complete recovery took place in about 4 weeks. Recovery time was scored as the number of days between hyperthermia and the day on which 50% of the motor function had returned. Irradiation (35 Gy) of a nerve segment, which included the heated part, resulted in a delayed recovery from the heat treatment compared to controls (heat only). The time interval and sequence between irradiation and hyperthermia hardly influenced the recovery delay. The size of the irradiated nerve segment did influence the recovery delay. Irradiation of a 20 mm nerve segment led to longer recovery delays than irradiation of a 10 mm segment (a delay of 5-10 days and 1-5 days respectively). A dose-response relation for the irradiation-induced delay in recovery was observed when a large segment (20 mm) of the nerve was irradiated immediately after heat with a dose ranging from 5 to 40 Gy. The delay in heat recovery was dose-dependent below 20 Gy, but after radiation doses above 20 Gy the recovery delay remained almost constant.

[Eleven years in a specialist practice dealing with reporting of occupational diseases can be rewarding]. / Elleve år i speciallaegepraksis med anmeldelse af arbejdsbetingede lidelser--det kan nytte.

During a period of 11 years in specialist practice, the authors notified 309 cases of occupationally-conditioned disease and occupational accidents. The majority of diseases were in the locomotor system. The authors reviewed the sex and age distributions and the distribution of the conditions involved. Their share of notifications from specialist practice during the period 1983-1986 was also reviewed together with the relevant specialties and the action taken by the Workers Supervision Authorities. The authors found their share in the notifications was relatively excessive but that it was declining because a great number of specialists have begun to notify conditions in the locomotor system. In addition, the authors have observed a considerable reduction in the rime taken to analyse cases by the Workers Supervision Authorities and an increase in the number of factory visits. The number of factory visits on account of the authors' notifications constituted 11% which shows that notification of occupationally-conditioned diseases is worth while.

Supernumerary ring chromosome 7 mosaicism: case report, investigation of the gene content, and delineation of the phenotype.

We report a girl with severe retardation of expressive speech development carrying a small, supernumerary ring chromosome derived from the proximal region of the long arm of chromosome 7. The r(7) chromosome is present in 50% of lymphocytes. We also review the six additional cases with a supernumerary r(7) chromosome reported in the literature. Among these patients, a severe retardation of productive language capabilities is seen as a shared clinical feature, irrespective of the degree of mosaicism as detected in blood. The dysmorphisms in these patients are minor and no shared congenital abnormalities seen. We, therefore, recommend chromosomal investigations in children with unexplained, disproportionately retarded expressive speech performance. Because speech and language acquisition are subject to genetic influences, we investigated whether there are genes on the r(7) chromosome that may affect brain development or function in a dosage-dependent manner. We found that both in our patient and in four patients described by others, the supernumerary r(7) chromosome contains the region from the centromere up to marker D7S613 located at 7q11.23. We speculate that the effects on speech acquisition are mediated by the supernumerary copies of the STX1A and LIMK1 genes, which are both located in this region and known to suppress neurite growth when overexpressed in vitro.

Embryonic development of Bithynia tentaculata L. (Prosobranchia, Gastropoda). I. Cleavage.

The cell-lineage of Bithynia tentaculata L. has been studied through the formation of the fourth quartet of micromeres, i.e., the 81-cell stage. Up to the 28-cell stage, the cleavage pattern is radially symmetrical, and the divisions take place synchronously in the quadrants. Bilateral symmetry is established by the formation of the mesentoblast 4d at the future dorsal side. A resting stage preceding the formation of 4d is lacking. The mesentoblast 4d is formed earlier than the other fourth quartet cells, and it divides twice before 4a-4c are formed. Bilateral symmetry in the molluscan cross becomes apparent by different divisions of the tipcell and the basal cell in the dorsal arm. Dorsoventrality is progressively corroborated by the divisions in the second and third quartets of micromeres, which take a different course in the A and B quadrants on the one side and in the C and D quadrants on the other side.

The morphogenetic significance of the first quartet micromeres for the development of the snailBithynia tentaculata.

The significance of the first quartet of micromeres for the morphogenesis ofBithynia - a polar lobe-forming gastropod-has been studied by deletion experiments. After removal of the whole first quartet at the 8-cell stage a dorsoventrally organized veliger larva is formed. Apparently, an interaction between the animal micromeres and a vegetal macromere, which is essential for the origin of a dorsoventral organization in equally cleaving gastropods, is not required in polar lobe forming eggs. It is concluded that in these eggs dorsoventrality is determined by segregation of the polar lobe. The embryos, in which the first quartet has been removed, never develop head structures. This indicates that the capacity to form head structures is restricted to the first quartet of micromeres. Deletion of a specific first quartet micromere (1a, 1b, 1c or 1d) showed, however, that the individual cells of this quartet are not strictly determined right from their origin. Frequently regulative development was observed after removal of individual first quartet cells.

Inulin measurement in serum and urine with an autoanalyser, corrected for glucose interference.

A method is described for the semi-automated measurement of inulin concentrations in serum and urine in the presence of glucose. The concentration of glucose is measured simultaneously and is used to correct for almost all "inulin-like" interferences. The inulin standard curve is linear over a wide range (5-2500 mg/l). Between-run precision is < 6% and recovery from spiked sera is 98%. This method offers the possibility of measuring glomerular filtration rate in patients with varying glucose concentrations (e.g. diabetics).

Pharmacokinetics of dextropropoxyphene in acute poisoning.

Dextropropoxyphene (DP) is a commonly used medicament for suicide attempts in Denmark. Death may occur from respiratory depression or cardiac arrest. Mechanical hyperventilation which induces hypocapnia seems to reduce the occurrence of cardiac complications. In an attempt to relate the clinical events to the plasma concentrations of DP and the major metabolite norpropoxyphene (NP) we studied patients with acute poisoning treated either for 48 h with induced hypocapnia by hyperventilation or under a conservative regime. Hypocapnia was found to lead to a significant increase in the plasma half-life of DP. Under conservative treatment the plasma half-life was 17.9 +/- 6.7 (S.D.) h (n = 6), while under induced hypocapnia the mean of values from 5 patients was 30.5 +/- 6.9 (S.D.) h. Maximum serum levels of DP and NP were, however, significantly higher in the intensively treated patients (n = 7) than in those treated conservatively (n = 9), though less marked for NP compared to DP (DP: 4.9 +/- 2.1/2.4 +/- 1.0 mumol/l, NP: 6.3 +/- 2.4/4.1 +/- 1.7 mumol/l). A concentration dependent renal clearance of NP was not demonstrable. Possible explanations are the following: 1) A change in disposition pattern blood/tissue of DP during hypocapnia. 2) A reduced metabolism DP to NP during hypocapnia. 3) A reduction in other routes of elimination.

Localization of morphogenetic determinants in a special cytoplasm present in the polar lobe ofBithynia tentaculata (Gastropoda).

In the first polar lobe ofBithynia eggs a special plasm, the vegetal body, is present. It consists of a cupshaped aggregate of small vesicles. Centrifugation of eggs prior to first cleavage may result in displacement of the vegetal body. In about 50% of thecentriguged eggs the vegetal body is found outside the polar lobe, in one of the blastomeres. Removal of the polar lobe from non-centrifuged eggs always leads to severe defects in development. When the lobe is removed from centrifuged eggs, however, about 50% of the eggs develop into normal embryos. It is concluded that the presence of the vegetal body in a blastomere suffices to ensure normal development and, hence, that the polar lobe-specific morphogenetic determinants are contained within the vegetal body.

On the use of a hospital information system in evaluating clinical care: a case report.

In this paper we describe, as an example, how we obtained the information needed to evaluate a newly introduced protocol for ordering X-rays for ankle trauma patients. Extensive use was made of available data and facilities of the hospital information system (HIS). Procedures for collecting the required additional data, which were not recorded in the HIS but were needed to evaluate the protocol, were embedded in the current medical and administrative routine of the emergency room. These additional data were also stored in the HIS. Periodically all data were downloaded to a personal computer to analyse the impact of using the protocol on quality of care and costs. In total 1241 patients entered the study, and for 1149 patients a complete dataset was obtained. The sensitivity and specificity of the protocol at the threshold value which was used during the initial study period was 0.77 and 0.80. The reduction in the number of ankle X-rays due to the protocol was significant when compared with a strategy of ordering an X-ray for every ankle trauma patient visiting the emergency room.

Characterization of partial trisomy 9p due to insertional translocation by chromosomal (micro)FISH.

We describe a family with an insertion 12;9 translocation occurring in a balanced form in a mother and two sons, but in an unbalanced form in the proband, resulting in trisomy of chromosome region 9p22-->9p24. The proband manifests typical features of trisomy 9p; the clinical signs were mental and growth retardation, microcephaly, epicanthus, low-set ears, micrognathia, clinodactyly and hypoplastic phalanges of the fifth fingers, hypoplasia or absence of toenails, and extremely small genitals. The GTG-banded findings were confirmed using (micro)FISH. Intriguingly, the mother and the two carrier sons exhibited major learning difficulties that were not present in the non-carrier sister of the mother: this may be due to a gene disruption or induction of abnormal expression. Dysmorphic features were not present in the three carriers. We compare our clinical and cytogenetic findings with other cases of partial trisomy 9p reported in the literature.

Immunocytochemical analysis of cisplatin-induced platinum-DNA adducts with double-fluorescence video microscopy.

To detect low-level DNA platination, a sensitive immunocyto- and histochemical technique was developed using a polyclonal antibody. The antibody GPt, derived after immunization of rabbits with highly platinated DNA and purified with affinity chromatography, detected the main platinum (Pt)-containing intrastrand and interstrand adducts. Double-fluorescence microscopy image analysis was used to quantify Pt-DNA adducts with Hoechst 33258 fluorescence to locate the nuclei and with fluorescein isothiocyanate fluorescence to measure the immunosignal. A two- to five-fold dose-dependent difference in the level of cisplatin (CDDP)-induced Pt-DNA adducts between a CDDP-sensitive and -resistant human tumour cell line was detected. Large differences in Pt-DNA adduct levels after in vitro CDDP incubation between human buccal cells, lymphocytes and biopsies of different tumour types were observed. Pt-DNA adduct levels were fivefold higher in human testicular tumours than in colon tumours, representing CDDP-sensitive and -resistant tumours, respectively, in the clinic. These data suggest the possibility of predictive testing by measuring Pt-DNA adduct levels. Pt-DNA adducts in patients after treatment with CDDP were shown in normal buccal cells and in imprints of fresh tumour biopsies as well as in paraffin-embedded tumour cells. The analysis of Pt-DNA adducts at a single-cell level in small samples of normal and tumour cells during and/or after treatment is feasible with GPt and will hopefully enable more selective treatment of patients.