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Rationale: It is well established that resting energy expenditure (REE) decreases with age. Data derived from indirect calorimetry (IC) are still limited with respect to the number of high aged individuals, BMI groups and health conditions. Therefore, IC generated REE of the BASAROT sample and those calculated according to the Harris-Benedict (HB) equation were used to re-evaluate the proposed association between REE and age. Methods: The IC-BASAROT sample combines the result of IC performed in 2622 individuals from 10 centers (7 Germany, 2 Italy, 1 Netherlands) done under strictly standardized conditions (e.g. at least 8h of fasting) in free-living, mostly healthy adults aged 18 to 100 years including all BMI ranges. IC was performed by canopy technique (Cosmed Quark RMR/Sensor Medics Vmax29) in 96.5% of cases and by face mask (Cosmed Fitmate) in 3.5%. Weight was measured by calibrated scales and height was determined to the nearest of 1mm. Results: REE in the total sample (BMI: 26.9±9.1 kg/m², 43.7±17.6 y) correlated more positively with body weight than with BMI (r=0.768; p

Growth factor-mediated signal transduction pathways in gynecologic malignancies are summarized. Targeting critical tyrosine kinase pathways may lead to more specific anticancer regimens in gynecologic oncology. During the past 10 years significant progress in cancer treatment has been made through the discovery of potent specific and well-tolerated inhibitors of signal transduction. Improved understanding of molecules involved in signal transduction pathways will undoubtedly result in an increasing number of compounds under clinical investigation. Some of the described molecular therapeutic approaches are suitable to enrich the conventional chemotherapy.

The developing condylar cartilage of the temporomandibular joint responds to changes in load by adaptive growth. Because local regulatory events taking place during growth processes are not well understood, investigation of extracellular matrix composition could provide new information about which matrix molecules are involved in the regulation of growth processes in this avascular tissue. The large chondroitin sulphate-rich proteoglycans in the mandibular condyle were compared to the proteoglycans in the weight-bearing femoral condyle of juvenile domestic pigs with respect to their buoyant density, chemical composition and immunological identity after isolation by dissociative extraction and CsCl density-gradient centrifugation. The distribution of these proteoglycans was studied in cryosections of mandibular condyle by immunohistochemistry using polyclonal antibodies produced against pig large proteoglycans. In the mandibular condyle, predominantly in the articular zone, the relative amount of proteoglycans with a low glycosaminoglycan content was greater than in femoral cartilage. The large proteoglycan immunologically related to aggrecan gave a protein core of 450 kDa after enzymatic deglycosylation and clearly possessed less keratan sulphate than in femoral aggrecan. Furthermore, the mandibular tissue contained another large proteoglycan with a protein core of 550 kDa after enzymatic deglycosylation, which was immunologically related to the fibroblast-like versican. Immunohistochemistry showed aggrecan increasing in amount inferiorly. In contrast, "versican' was exclusively found in the fibrous and differentiation layers. Aggrecan is mainly responsible for shock absorption and versican and its homologues may be involved in the control of cell proliferation and differentiation. Thus the matrix components of the mandibular condyle seem to be adapted to its special functional needs including parallel articulation and growth.

Age-related changes in the relative proportion of high buoyant density proteoglycans of the soft tissue cover of the mandibular condyle of domestic pigs and the chemical composition of these proteoglycans were investigated by biochemical methods. The relative proportion of high buoyant density proteoglycans has been shown to decrease with age, with their chemical composition varying as follows: With age, the relative proportions of protein increased, galactosamine decreased, glucosamine was similar in newborns and juveniles, but clearly increased in adults, galactosamine plus glucosamine decreased. As measured by 1H-NMR-relaxation spectroscopic methods, the concentration of free tissue water of the soft tissue cover of the mandibular condyle decreased with age. Its mobility, however, was greater in newborns than in juveniles, but similar in juveniles and adults. The concentration of free tissue water, but not its mobility, correlated weakly with relative proportions of polyanionic side chains (e.g. chondroitin sulphate plus keratan sulphate chains) of the high buoyant density proteoglycans. By affecting the concentration of free tissue water and in turn of matrix solutes and cellular environments, the high buoyant density proteoglycans of the soft tissue cover of the mandibular condyle seem to be involved in regulation of age-related changes in the adaptive remodelling activity of the mandibular condyle.

Patients with chronic kidney disease (CKD) show a broad spectrum of clinical symptoms intimately related to the disturbed mineral and bone metabolism and summarized as CKD-mineral-bone disorder (CKD-MBD). Whereas in adults an impaired bone metabolism translates mainly into an increased risk of fractures, in pediatric CKD patients rickets, skeletal deformations, and severe growth failure are additional and severe clinical findings. Further-more, an elevated Ca x P ion product, secondary hyperparathyroidism (sHPT) as well as concomitant calcitriol medication have been linked to ectopic (vascular) calcification, in which is strongly associated with the dramatically high cardiovascular morbidity and mortality even in pediatric CKD patients. Thus, in these patients the impaired mineral metabolism is the link between skeletal and cardiovascular disease. In other words, the complex interplay between kidney, skeleton, parathyroid gland, the intestine and the cardiovasculature is severely disturbed in CKD. This review summarizes the recent findings in our understanding of bone cell biology and alterations of mineral metabolism in children with CKD.

Adenovirus (ADV)-mediated gene therapy with the thymidine kinase (TK) gene under control of the Rous sarcoma virus (RSV) promotor followed by the administration of acyclovir has been established in vitro for the treatment of ovarian cancer cells and has been used as the basis for intraperitoneal phase I clinical trials. It is unclear how long a significant degree of transgene translation can be expected after adenovirus-mediated TK transduction, where the transcriptional complex is localized in the nucleus in an episomal fashion and thus without stable integration. The possible interaction of acyclovir pretreatment with subsequent ADV-RSV-TK transduction also remains to be elucidated. Transgene expression and cell killing efficacy were analysed based on multiplicity of infection (MOI) and MTT assay. Anti-TK-antibody 1397 was used for immunocytochemistry and Western blot analysis of TK expression. After transduction with ADV-RSV-TK at an MOI of 66, TK translation increased strongly in MDH 2774 and OVCAR-3 cell lines during the initial 48 hours. Virtually constant expression of the TK transgene was observed by Western blot during eight days. Cell killing efficacy was increased by repeated daily administrations of acyclovir. Pretreatment with acyclovir did not result in significantly increased cell killing efficacy. No negative effect of acyclovir on ADV-RSV-TK transduction was observed. The at least week-long expression of the TK transgene with persistently increasing efficacy of cell killing after ADV-mediated tumor cell transduction provide a realistic basis for the development of multicycle ADV-mediated TK gene therapy approaches in the treatment of ovarian cancer. Continuous i.v. acyclovir treatment or daily oral acyclovir-prodrug therapy might simplify the substrate regimen for the TK gene.

Alternative splicing represents an important nuclear mechanism in the post-transcriptional regulation of gene expression, which is frequently altered during tumorigenesis. Previously, we have described marked changes in alternative splicing of the CD44 gene in ovarian and breast cancer. In the latter one we described also a specific induction of splicing factors during tumor development. Now we have focussed our studies on the expression profiles of splicing factors, including classical SR proteins, Tra2 and YB-1 in physiological and malignant ovarian tissues by RT-PCR and Western blot analysis. We detected changed expression pattern with higher levels of phosphorylated 30 kDa SR proteins as well as relatively high concentrations of hyperphosphorylated Tra2 protein isoforms in ovarian cancer. RT-PCR analysis revealed a marked induction of SC35 and ASF/SF2 as well as mRNA levels in malignant ovarian tissue. These results suggest gene-specific alterations of expression rather than a general induction of the splicing machinery. Together with previously performed functional studies of CD44 splicing these findings implicate that altered expression profiles of SR proteins, Tra2beta and YB-1 might be responsible for the known changes of alternative CD44 splicing in ovarian cancer.

Adrenomedullin (ADM) is an angiogenic factor that has also been shown to be a mitogen and a hypoxia survival factor for tumour cells. These properties point to ADM as a potential promoter of human malignancies, but little data are available concerning the expression of ADM in human breast cancer. In the present work, we have examined ADM peptide expression in a series of malignant breast tumours by immunohistochemistry using a newly developed anti-ADM monoclonal antibody. In addition, ADM plasma concentrations in breast cancer patients and healthy controls were determined by radioimmunoassay. Of the examined breast cancer samples, 27/33 (82%) showed a moderate to strong staining intensity. ADM-peptide expression in breast tumours was significantly correlated with axillary lymph node metastasis (P=0.030). Analysis of ADM plasma concentrations showed no significant difference between the circulating ADM levels of breast cancer patients and healthy controls. However, a significant positive correlation was found between tumour size and plasma ADM levels (r=0.641, P=0.017). Moreover, ADM levels in breast cancer patients correlated with the presence of lymph node metastasis (P=0.002). In conclusion, we have shown for the first time that ADM peptide is widely expressed in breast cancer and that the degree of expression is associated with lymph node metastasis. ADM peptide in plasma of breast cancer patients reflects the size of the primary tumour, but is unlikely to be a useful tumour marker for the detection of breast cancer. Plasma ADM might represent an independent predictor of lymph node metastasis. The clinical implications of these findings remain to be evaluated.

With the widespread use of routine abdominal ultrasound examination during pregnancy, adnexal masses are observed with increasing frequency. Most patients are clinically asymptomatic at the time of presentation, and most of the adnexal masses detected during early pregnancy disappear during the first 16 weeks of pregnancy. Ovarian tumors are estimated to occur in about 1 in 1,000 pregnancies and of these 3% are malignant. Here we present an overview about frequency, diagnostic procedures and pathological characteristics of these ovarian tumors. Moreover, current modalities for treatment during pregnancy are summarized. Surgical treatment of the adnexal masses has to be performed with adequate staging and debulking equal to the treatment of non-pregnant women. However, whereas during organogenesis abortion has to be considered prior to chemotherapy, later in pregnancy surgical debulking as complete as possible, followed by taxol-platinum chemotherapy is indicated. If the fetus is not viable at the time of primary surgery, neoadjuvant chemotherapy and complementation of surgery after delivery of the baby should be performed. It should be stressed that chemotherapy for ovarian cancer applied during pregnancy appears to be safe. However, no studies have evaluated the long-term consequences for children exposed to intra-uterine chemotherapy. Aspiration of cysts should be avoided, as the correlation between the histological evaluation of an ovarian malignancy and the cytological evaluation of aspirates is poor. Moreover, spillage of malignant cysts is hazardous for the patient.

Ten patients with recurrent ovarian cancer received a combined treatment of optimal tumor debulking, adenovirus-mediated herpes simplex virus thymidine kinase gene therapy (GT), and systemic application of acyclovir or valacyclovir and topotecan. Biopsies were taken at the time of secondary debulking about 1 month after the application of GT and chemotherapy and were analyzed for expression of coxsackie-adenovirus receptor (CAR) and integrins alphavbeta3 and alphavbeta5 with respect to treatment response.Treatment modalities and study design have been described recently. Immunohistochemistry was used to visualize expression of CAR and integrins alphavbeta3 and alphavbeta5 in tumor samples taken before and after application of GT.Before GT six of ten patients presented with CAR-positive and four with CAR-negative tumors. After GT all tumors showed CAR expression. Integrin alphavbeta3 was found in all tumors before and after GT. Expression of integrin alphavbeta5 was seen in eight of ten tumor samples before GT and in all samples after GT.Despite the importance of CAR and integrin expression for successful adenovirus internalization, other cell surface receptors might be involved in this process. It is too early to decide whether expressions of CAR and integrin alphavbeta3/alphavbeta5 on tumor cells are appropriate additional inclusion criteria for the enrollment of patients in GT trials. Further research is necessary to evaluate the effect of GT plus chemotherapy on CAR and integrin expression.

To elucidate the function of keratan sulfate proteoglycan (KS-PG) in the human uterine cervix, we analyzed its distribution with respect to physiologic conditions.Immunohistochemistry was used to localize KS bearing proteoglycans (mAb 5D4) and decorin (mAb 6B6) in the lower uterine segment. Proteins present in cervical mucous were labeled with biotin, glycosaminoglycan chains were digested enzymatically, and the samples were analyzed by Western blot.Decorin was detected throughout the extracellular matrix, in tissues from menstruating nonpregnant women, in early pregnancy, from women who had cesarean at term, at postpartum hysterectomy, and from postmenopausal women. In menstruating nonpregnant women, in early pregnancy (first trimester), and in postmenopausal women, KS-PG was detectable only in epithelial, mucous-producing cells. Interestingly, in samples obtained either at the time of cesarean at term (lower uterine segment) or after postpartal hysterectomy, KS-PG was detectable throughout the extracellular matrix, indicating that the expression of KS-PG is associated with reorganization of the tissue. Biochemical analysis of the KS present in mucous revealed a core protein in the range of 220 kDa, suggesting an identity with the large KS-PG described previously.At parturition, a large KS-PG, which is virtually exclusively present in the cervical mucous of either early or nonpregnant women, was detected in the extracellular matrix. This finding indicates that cervical ripening is accompanied not only by quantitative but also by qualitative changes in the composition of the extracellular matrix.

Macular corneal dystrophy can be classified into three different immunophenotypes according to presence and distribution of sulfated keratan sulfate (SKS) in the cornea and serum levels of SKS. Immunophenotype I A has yet only been reported in patients with macular corneal dystrophy from Saudi Arabia.Case report of a patient with macular corneal dystrophy type I A from Germany. Indirect immunohistochemistry for sulfated keratan sulfate was performed on the corneal buttons obtained after perforating keratoplasty (monoclonal antibodies 3D12/H7 and 5-D-4). The serum concentration of SKS was determined using the monoclonal antibody 5-D-4 in a serum antigen-inhibition assay. Immunogold labeling with the 5-D-4 antibody was performed to allow ultrastructural localization of SKS in the cornea.The patient demonstrated the typical clinical picture of macular corneal dystrophy. Serum levels of SKS were not detectable. Positive immunohistochemistry for SKS was found only within single keratocytes of both corneae without staining of the stroma (3D12/H7 and 5-D-4). Immunogold labeling localized SKS within stromal keratocytes and proved this patient to suffer from macular corneal dystrophy immunophenotype I A.Macular corneal dystrophy immunophenotype I A can be found outside Saudi Arabia in a German patient. All three immunophenotypes of macular corneal dystrophy are present in corneal buttons from German patients.

Reference hematologic and plasma chemistry values were determined from 103 blood samples collected from 53 clinically healthy brown tree snakes (Boiga irregularis). Female snakes had significantly higher mean plasma values for total solids, total protein, calcium (Ca), phosphorus (P), uric acid, and blood monocyte percentage than did males, whereas males had significantly higher mean plasma fibrinogen values. The variances for hematocrit, monocyte percentage, azurophil percentage, plasma total solids, plasma total protein, albumin, Ca, and P also differed significantly between sexes. The higher mean values and greater variances for plasma total protein, plasma total solids, Ca, and P in the female snakes were probably associated with yolk synthesis and accumulation.

We have previously shown that human articular chondrocytes synthesize large amounts of interleukin-6 (IL-6) upon stimulation with proinflammatory cytokines and that they express the IL-6 receptor. The present study was undertaken to analyze whether different IL-6-type cytokines can induce synthesis of the acute-phase protein alpha1-antitrypsin in human articular chondrocytes.Chondrocytes from human articular cartilage, cultured in agarose, were stimulated with IL-6-type cytokines. Total RNA was isolated and analyzed by Northern blotting. Levels of alpha1-antitrypsin protein were determined by enzyme immunoassay.Stimulation of chondrocytes with oncostatin M (OSM) and IL-6 led to a 5-10-fold increase in alpha1-antitrypsin synthesis. This increase was dose and time dependent. Furthermore, OSM and IL-6 induced IL-6 synthesis in chondrocytes, resulting in an autocrine amplification loop.Our data strongly suggest the existence of a local acute-phase response in the joint. Synthesis of the acute-phase protein alpha1-antitrypsin, a major inhibitor of serine proteinases, may be an important protective mechanism of articular chondrocytes to prevent cartilage damage in inflammatory joint diseases.

The disposition of drugs may be influenced by hyperbaric conditions, in particular by changes of liver perfusion. The effect of hyperbaric hyperoxia on the pharmacokinetics of lidocaine, a drug eliminated in the liver with a perfusion-limited clearance, was investigated in human volunteers in a crossover trial.A single dose lidocaine i.v. bolus (0.69 or 0.75 mg x kg(-1)) was administered to two volunteers under normobaric conditions (NB: 1 bar or 0.1 MPa, air) and under hyperbaric/hyperoxic conditions (HBO: 2.5 bar or 0.25 MPa, alternating 100% O2-breathing for 20 min and air breathing for 5 min). Blood samples were serially collected for 5 h (NB) or 75 min (HBO), and lidocaine concentration in serum was measured by immunoassay. Data were analyzed assuming linear kinetics and an open two-compartment model.At 1 bar or 0.1 MPa, lidocaine injection caused only slight dizziness and buzzing in the ear. Heart rate and blood pressure were not influenced. Under HBO, lidocaine injection caused marked dizziness and buzzing in the ears, sweating, tremor and coordination-disturbances, even though maximal lidocaine concentrations (0.63 mg x L(-1) and 0.70 mg x L(-1)) were far below therapeutic serum concentrations (1.5-5.0 mg x L(-1)). Pharmacokinetic parameters of lidocaine were similar to those published earlier (T1/2beta: 110+/-16 min; CI: 12.6+/-2.9 ml x min(-1) x kg(-1); Vss: 1.73+/-0.18 L x kg(-1)). There was no indication for effects of HBO on the disposition of lidocaine (p0.05).The pharmacokinetics of lidocaine do not seem to be influenced in a clinically relevant way in humans by a single HBO-exposure under usual therapeutic conditions. Side effects of lidocaine at 2.5 bar or 0.25 MPa may be caused by pharmacodynamic interactions between lidocaine and hyperbaric/hyperoxic conditions.

To determine the concentrations of tumor necrosis factor (TNF)alpha, interleukin (IL)beta, IL-6 and IL-8 and the expression of endothelial leukocyte adhesion molecule-1 (ELAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in the lower uterine segment during parturition at term and to investigate correlations with the stage of cervical dilatation and with the duration of labor.Biopsy specimens were taken from the lower uterine segment of 59 women undergoing non-elective cesarean sectin at term at different stages of cervical dilatation (2 cm, n = 17; 2-3 cm, n = 12; 4-6 cm, n = 13;6 cm, n = 17) after varying length of the time in labor (1-6 h, n = 18;6-12 h, n = 16;12-24 h, n = 15;24 h, n = 10). The concentrations of TNF alpha, IL-1 beta, IL-6 and IL-8 in protein extracts of these specimens were determined by enzyme linked immunosorbent assays and the expression of ELAM-1 and VCAM-1 was studied by immunohistochemistry. Staining was considered negative if5% of the endothelial cells were stained and positive ifor = 5% were stained.At2 cm dilated cervix the median concentrations of TNF alpha, IL-1 beta, IL-6 and IL-8 were 8.0 pg/mg total protein (TP), 1.3 pg/mg TP, 12.6 pg/mg TP and 17.2 pg/mg TP, respectively. At 4-6 cm dilatation the median concentrations were increased (TNF alpha: 22.2 pg/mg TP [P = 0.05], IL-1 beta: 22.2 pg/mg TP [P0.05], IL-6: 1226 pg/mg TP [P0.05], IL-8: 2081 mg/mg TP [P0.05]). At6 cm dilatation no further increase was observed. The expression of ELAM-1 and VCAM-1 was significantly greater at6 cm dilatation than2 cm dilatation (P0.001 and 0.05, respectively). With increasing duration of labor, the concentrations of IL-1 beta, IL-6 and IL-8 (but not of TNF alpha) increased and were significantly higher at6 cm dilatation than at2 cm (P0.05). Expression of ELAM-1 and VCAM-1 did not vary with duration of labor.These first investigations of cytokines and cell adhesion molecules in the lower uterine segment at both different stages of cervical dilatation and different duration of labor support the decisive role of cytokines for normal cervical dilatation. Local changes in the lower uterine segment independent from the effects of labor (e.g. increase of endothelial adhesivity) seem to be of special importance in this process.