Exercise affects symptom severity but not biological measures in depression and somatization – Results on IL-6, neopterin,tryptophan, kynurenine and 5-HIAA

Exercise leads to symptom reduction in affective disorders and functional somatic syndromes. Biological hypotheses of underlying mechanisms include serotonergic and immunological pathways. We aimed to investigate biological features in persons with major depression and somatoform syndromes, and to analyze effects of short-term graded exercise on these parameters. Baseline values for depressive and somatoform symptoms, tryptophan, kynurenine, 5-hydroxyindoleacetic acid, neopterin and interleukin-6 were compared with those after one week of increased and one week of reduced physical activity. Thirty-eight persons with major depression, 27 persons with a minimum of 6–8 somatoform symptoms, and 48 healthy controls participated in the study. Depressive and somatoform symptoms were reduced after the active week, and an interaction pointed towards group-specific reduction of psychopathology. Participants with major depression had lower levels of kynurenine compared to controls, with intermediate concentrations in somatoform patients. There were no systematic associations of symptom improvement with biological changes. A possible limitation of the design is that a control condition with low physical activity, but no placebo condition was included. People with multiple somatoform symptoms and major depression benefit from a short and low-graded exercise intervention. These effects do not seem to be mediated by changes in serotonergic and inflammatory parameters.     

Hemocompatibility of a Miniaturized Extracorporeal Membrane Oxygenation and a Pumpless Interventional Lung Assist in Experimental Lung Injury

Extracorporeal membrane oxygenation (ECMO) is used for most severe acute respiratory distress syndrome cases in specialized centers. Hemocompatibility of devices depends on the size and modification of blood contacting surfaces as well as blood flow rates. An interventional lung assist using arteriovenous perfusion of a low-resistance oxygenator without a blood pump (Novalung, Hechingen, Germany) or a miniaturized ECMO with reduced filling volume and a diagonal blood pump (Deltastream, Medos AG, Stolberg, Germany) could optimize hemocompatibility. The aim of the study was to compare hemocompatibility with conventional ECMO. Female pigs were connected to extracorporeal circulation for 24 h after lavage induced lung injury (eight per group). Activation of coagulation and immune system as well as blood cell damage was measured. A P value <0.05 was considered significant. Plasmatic coagulation was slightly activated in all groups demonstrated by increased thrombin-anti-thrombin III-complex. No clinical signs of bleeding or thromboembolism occurred. Thrombelastography revealed decreased clotting capacities after miniaturized ECMO, probably due to significantly reduced platelet count. These resulted in reduced dosage of intravenous heparin. Scanning electron microscopy of oxygenator fibers showed significantly increased binding and shape change of platelets after interventional lung assist. In all groups, hemolysis remained negligible, indicated by low plasma hemoglobin concentration. Interleukin 8 and tumor necrosis factor-α concentration as well as leukocyte count remained unchanged. Both devices demonstrated adequate hemocompatibility for safe clinical application, although a missing blood pump did not increase hemocompatibility. Further studies seem necessary to analyze the influence of different blood pumps on platelet drop systematically.     

A possible mechanism for exonuclease 1-independent eukaryotic mismatch repair

Mismatch repair contributes to genetic stability, and inactivation of the mammalian pathway leads to tumor development. Mismatch correction occurs by an excision-repair mechanism and has been shown to depend on the 5′ to 3′ hydrolytic activity exonuclease 1 (Exo1) in eukaryotic cells. However, genetic and biochemical studies have indicated that one or more Exo1-independent modes of mismatch repair also exist. We have analyzed repair of nicked circular heteroduplex DNA in extracts of Exo1-deficient mouse embryo fibroblast cells. Exo1-independent repair under these conditions is MutLα-dependent and requires functional integrity of the MutLα endonuclease metal-binding motif. In contrast to the Exo1-dependent reaction, we have been unable to detect a gapped excision intermediate in Exo1-deficient extracts when repair DNA synthesis is blocked. A possible explanation for this finding has been provided by analysis of a purified system comprised of MutSα, MutLα, replication factor C, proliferating cell nuclear antigen, replication protein A, and DNA polymerase δ that supports Exo1-independent repair in vitro. Repair in this system depends on MutLα incision of the nicked heteroduplex strand and dNTP-dependent synthesis-driven displacement of a DNA segment spanning the mismatch. Such a mechanism may account, at least in part, for the Exo1-independent repair that occurs in eukaryotic cells, and hence the modest cancer predisposition of Exo1-deficient mammalian cells.     

Specific immunoadsorption of pathogenic autoantibodies in pemphigus requires the entire ectodomains of desmogleins

Pemphigus foliaceus (PF) and pemphigus vulgaris (PV) are life‐threatening autoimmune blistering skin diseases. They are characterized by circulating autoantibodies which bind to the ectodomains of desmoglein (Dsg) 1 and Dsg3. These antibodies induce acantholysis in skin and mucous membranes. In severe cases of pemphigus, immunoadsorption is applied to remove total IgG from patient plasma using protein A or other ligands. To develop a specific adsorber for anti‐Dsg antibodies, epitope mapping studies of Dsg1 and Dsg3 ectodomains were conducted. Dsg variants were expressed on the surface of HEK‐293 cells and analysed for reactivity with pemphigus and control sera by indirect immunofluorescence technique. For Dsg1, a construct consisting of domain 1 directly fused to domain 5, seemed to be suitable for specific immunoadsorption of anti‐Dsg1 antibodies. The recognized epitopes were mainly conformation‐dependent. However, adsorption of pemphigus foliaceus IgG using this protein coupled to a Sepharose matrix did not completely remove pathogenicity from the sera, as proven by a keratinocyte dissociation assay. In contrast, full‐length Dsg1 and Dsg3 ectodomains were able to specifically adsorb anti‐Dsg antibodies and to efficiently eliminate pathogenicity. Therefore, the complete and correctly folded ectodomains of both desmogleins are required for therapeutic immunoadsorption.     

Immediate versus conventional loading of palatal implants in humans: a first report of a multicenter RCT

This study aims to analyze the clinical performance of two loading concepts on second-generation palatal implants (Orthosystem, Straumann, Basel, Switzerland) in a prospective multicenter randomized controlled clinical trial. At the time of this interim analysis, 41 patients have been randomized on a 1:1 basis to one of two treatment groups. Group 1 underwent conventional loading of palatal implants after a healing period of 12 weeks (gold standard) while group 2 underwent immediate implant loading within 1 week after implant insertion. We report initial results at 6 months after functional loading. The primary outcome parameter was implant success (no implant mobility, no implant loss). The implants in both groups were initially stable at the time of insertion, and all were eligible for randomization. Twenty-two patients (group 1) were subjected to conventional implant loading after 12 weeks while 19 patients (group 2) received immediate functional loading within the first week after insertion. Direct (e.g. distal jet appliances) as well as indirect forms of anchorage (conventional or modified transpalatal arch) were used. The magnitude of orthodontic forces ranged between 1 and 4 N for the immediate loading group and between 1 and 5 N for the conventional loading group. One implant in group 1 was lost during the healing phase. One dropout was registered in group 2. Thirty-nine implants were functionally loaded for over 6 months now. These preliminary data provide first evidence of the fact that immediate loading of palatal implants yields equivalent success rates as conventional loading to 4 N after 6 months.     

Cisplatin-resistant neuroblastoma cells express enhanced levels of epidermal growth factor receptor (EGFR) and are sensitive to treatment with EGFR-specific toxins

PURPOSE: Neuroblastomas frequently show expression of the epidermal growth factor receptor (EGFR) and may therefore be susceptible to EGFR-targeted therapies. Here, EGFR expression and functionality was investigated in parental chemosensitive neuroblastoma cell lines (UKF-NB-3, IMR-32, NLF, SH-SY5Y) and their cisplatin-resistant sublines (UKF-NB-3(r)CDDP(1000), IMR-32(r)CDDP(1000), NLF(r)CDDP(1000), and SH-SY5Y(r)CDDP(500)). Moreover, the EGFR antibody cetuximab, the EGFR tyrosine kinase inhibitor Tyrphostin B46, and recombinant EGFR-targeted toxins were investigated for their influence on the viability and growth of neuroblastoma cells. EXPERIMENTAL DESIGN: EGFR expression and function was measured by flow cytometry or Western blot. Cell viability was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis was examined by immunostaining for active caspase-3 or cleaved poly(ADP-ribose) polymerase. Cellular binding of FITC-labeled immunotoxins was studied by flow cytometry, and cellular uptake was studied by confocal laser scanning microscopy. RESULTS: The EGFR-targeted antibody and growth factor toxins scFv(14E1)- Pseudomonas exotoxin A (ETA) and TGF-alpha-ETA exerted anti-cancer effects in neuroblastoma cell lines that were insensitive to cetuximab or EGFR tyrosine kinase inhibitors. Furthermore, adaptation of chemosensitive neuroblastoma cells to cisplatin increased EGFR expression and sensitivity to both recombinant toxins. Treatment of chemosensitive neuroblastoma cells with cisplatin reversibly increased EGFR expression, whereas cisplatin-resistant cells showed enhanced EGFR expression independent of the presence of cisplatin. Combination treatment with scFv(14E1)-ETA or TGF-alpha-ETA and cisplatin exerted significantly improved anticancer effects compared with either single treatment in parental neuroblastoma cells, cisplatin-resistant sublines, and primary cultures. CONCLUSIONS: EGFR-targeted cytotoxic reagents such as scFv(14E1)-ETA and TGF-alpha-ETA represent promising candidates for further development as antineuroblastoma agents, especially in combination with cisplatin.     

The AA genotype of the regulatory BCL2 promoter polymorphism ( 938C>A) is associated with a favorable outcome in lymph node negative invasive breast cancer patients.

PURPOSE: Expression of the antiapoptotic and antiproliferative protein Bcl-2 has been repeatedly shown to be associated with better clinical outcome in breast cancer. We recently showed a novel regulatory (-938C>A) single-nucleotide polymorphism (SNP) in the inhibitory P2 BCL2 gene promoter generating significantly different BCL2 promoter activities. EXPERIMENTAL DESIGN: Paraffin-embedded neoplastic and nonneoplastic tissues from 274 patients (161 still alive after a follow-up period of at least 80 months) with primary unilateral invasive breast carcinoma were investigated. Bcl-2 expression of tumor cells was shown by immunohistochemistry; nonneoplastic tissues were used for genotyping. Both the Bcl-2 expression and the (-938C>A) genotypes were correlated with the patients' survival. RESULTS: Kaplan-Meier curves revealed a significant association of the AA genotype with increased survival (P = 0.030) in lymph node-negative breast cancer patients, whereas no genotype effect could be observed in lymph node-positive cases. Ten-year survival rates were 88.6% for the AA genotype, 78.4% for the AC genotype, and 65.8% for the CC genotype. Multivariable Cox regression identified the BCL2 (-938CC) genotype as an independent prognostic factor for cancer-related death in lymph node-negative breast carcinoma patients (hazard ratio, 3.59; P = 0.032). Immunohistochemical Bcl-2 expression was significantly associated with the clinical outcome of lymph node-positive but not of lymph node-negative breast cancer patients. In lymph node-negative cases, the (-938C>A) SNP was both significantly related with the immunohistochemically determined level of Bcl-2 expression (P = 0.044) and the survival of patients with Bcl-2-expressing carcinomas (P = 0.006). CONCLUSIONS: These results suggest the (-938C>A) polymorphism as a survival prognosticator as well as indicator of a high-risk group within patients with lymph node-negative breast cancer.     

Tamoxifen: evidence by 32P-postlabeling and use of metabolic inhibitors for two distinct pathways leading to mouse hepatic DNA adduct formation and identification of 4-hydroxytamoxifen as a proximate metabolite

Exposure to pentachlorophenol (PCP) strongly intensifies theformation of mouse hepatic DNA adducts elicited by oral administrationof tamoxifen (TAM), as previously shown by ³²P-postlabeling.To explain this effect, PCP was proposed to interfere with thedetoxication by sulfate conjugation of an as yet unidentifiedhydroxylated proximate TAM metabolite. A comparison of the presentand earlier results shows that the hepatic TAM adduct patternin female ICR mice depended on the route of administration ofTAM (120 µmol/kg), with oral administration primarilyeliciting formation of more polar adducts (termed group I adducts),while after i.p. administration less polar adducts (group II)predominated over group I adducts by a factor of 17.5. All theseadducts were also formed in female Sprague–Dawley ratsafter i.p. dosing with TAM, but total adduct levels were 3.5-to 5-fold higher than in mice. After four daily i.p. treatments,TAM adducts accumulated in mouse liver DNA in a non-linear fashion.Adduct levels were 30–50 times lower in mouse kidney andlung than in liver. The phenolic metabolite 4-hydroxy TAM (120µimol/kg) exclusively led to formation of polar (groupI) hepatic adducts, and this process was stimulated 8-fold bycoadministration of PCP (75 µimol/kg). Co-administrationof PCP with the parent compound led to an 11-fold enhancementof group I adduct formation; simultaneously, levels of groupII adducts were suppressed 6-fold. Another inhibitor of sulfateconjugation, 2,6-dichloro-4-nitrophenol, unlike PCP, had noeffect on group I adducts, but it reduced group II adduct formation2.2-fold. The PCP metabolite 2,3,5,6-tetrachlorohydroquinone(75 µimol/kg) did not significantly affect any major TAMadduct, suggesting that PCP itself was the active compound.Similar to group II TAM adducts, the formation of hepatic safrole–DNAadducts was inhibited in female ICR mice by both sulfotransferaseinhibitors, consistent with the proposal that metabolic