Adenovirus death protein (ADP) is required for lytic infection of human lymphocytes.

The adenovirus death protein (ADP) is expressed at late times during a lytic infection of species C adenoviruses. ADP promotes the release of progeny virus by accelerating the lysis and death of the host cell. Since some human lymphocytes survive while maintaining a persistent infection with species C adenovirus, we compared ADP expression in these cells with ADP expression in lymphocytes that proceed with a lytic infection. Levels of ADP were low in KE37 and BJAB cells, which support a persistent infection. In contrast, levels of ADP mRNA and protein were higher in Jurkat cells, which proceed with a lytic infection. Epithelial cells infected with an ADP-overexpressing virus died more quickly than epithelial cells infected with an ADP-deleted virus. However, KE37, and BJAB cells remained viable after infection with the ADP-overexpressing virus. Although the levels of ADP mRNA increased in KE37 and BJAB cells infected with the ADP-overexpressing virus, the fraction of cells with detectable ADP was unchanged, suggesting that the control of ADP expression differs between epithelial and lymphocytic cells. When infected with an ADP-deleted adenovirus, Jurkat cells survived and maintained viral DNA for greater than 1 month. These findings are consistent with the notion that the level of ADP expression determines whether lymphocytic cells proceed with a lytic or a persistent adenovirus infection.

TLR2 down-regulates FcεRI and its transcription factor PU.1 in human Langerhans cells.

BACKGROUND: Epidermal Langerhans cells (LC) expressing the high-affinity receptor for IgE (FcεRI) play a key role in atopic dermatitis (AD). AD skin is highly colonized with Staphylococcus aureus (S.a.), which are sensed by Toll-like receptor 2 (TLR2). We hypothesized that TLR2 may impact on the expression of FcεRI on LC. OBJECTIVES: To study a putative impact of TLR2 signaling on FcεRI, we analyzed FcεRI and known transcription factors of the receptor after ligand binding to TLR2. METHODS: We generated LC from CD34(+) progenitors in vitro (CD34LC) expressing FcεRI and TLR2 as well as its partners TLR1 and TLR6. The expression of FcεRI and known transcription factors of the receptor was analyzed on the protein and RNA level by flow cytometry, Western blotting, and real-time PCR. RESULTS: For CD34LC from 123 donors, we observed a high heterogeneity in FcεRI surface expression correlating with mRNA level of its α-chain. Stimulation of TLR1/2 or TLR2/6 dramatically down-regulated FcεRI on protein and mRNA level of both α- and γ-chain. Further analysis of putative transcription factors for FCER1A revealed the lack of GATA1 in CD34LC, weak expression of ELF1 and YY1, and high expression of PU.1. While ELF1 and YY1 appeared to be little affected by TLR2 engagement, PU.1 was significantly down-regulated. CONCLUSIONS: Taken together, our findings show that in human, LC ligation of TLR2 by S.a.-derived products down-regulates FcεRI and its transcription factor PU.1, thus suggesting that FcεRI is controlled by PU.1 in these cells.

Suppression of MAP kinases inhibits microglial activation and attenuates neuronal cell death induced by alpha-synuclein protofibrils.

Alpha-Synuclein (alpha-Syn) accounts, as a major component of Lewy bodies (LB), for the filamentous deposits in many cases of neurodegenerative diseases. Yet, little is known about the molecular mechanisms of neuronal loss in these diseases. The correlation between alpha-Syn oligomerization/aggregation and pathologies raises the key question of which molecular form of alpha-Syn (i.e. monomeric alpha-Syn, protofibrils or mature fibrils) represents the damage-inducing culprit in the scenario of synucleinopathies. We show that human alpha-Syn protofibrils (PFs) are potent activators of parallel proinflammatory signalling pathways (p38 and ERK1/2 MAP kinases and NF-kappaB) in microglial cells in vitro. Furthermore, stereotactic injection of alpha-Syn PFs into the substantia nigra of adult rats leads to a profound activation of microglia and adjacent neuronal cell loss, which can be attenuated by the MAP kinase inhibitor semapimod. We propose that the neurodegenerative process of alpha-synucleinopathies involves microglial activation through alpha-Syn released or extruded from cells with pathogenic alpha-Syn metabolism. Compounds that inhibit the MAPK/NF-kappaB pathways might be a promising pharmacological strategy for the treatment of the inflammatory component of synucleinopathies including PD.

[Efficacy of an intervention programme on burnout symptoms of partners of depressed patients]. / Wirksamkeit eines Interventionsprogramms auf die Burnout-Belastung von Lebenspartnern depressiv Erkrankter.

AIM OF THE STUDY: A newly developed group intervention programme was evaluated with regard to its effectiveness to decrease the burnout symptoms of the partners of depressed patients. METHODS: Within a period of six months, a group of 66 persons has taken part in the intervention for a total of twelve group sessions. A control group consisted of 50 persons without any intervention. Burnout was assessed using the German version of the Maslach Burnout Inventory (MBI). To evaluate the time effect, the burnout dimensions were used as independent variables in random effects models. RESULTS: Over the analyzed period of time no significant positive effect was measured on any of the assessed burnout dimensions. CONCLUSIONS: An increased inclusion of depressed patients in the intervention as well as an increased intensity and a lowering of the admission threshold for the heavily burdened relatives could increase the effectiveness of the program.

Multimerization of thyroglobulin (TG) during extracellular storage: isolation of highly cross-linked TG from human thyroids.

Thyroglobulin (TG) is the major soluble protein of the thyroid and is known to be extracellularly stored for future liberation of thyroid hormones. We have developed techniques for the isolation of an insoluble storage form of human TG present in the follicle lumen. The application of these techniques yielded insoluble and translucent colloid globules varying in size (50-500 microns) and shape and consisting primarily of densely packed TG. Intact colloid globules exhibited the imprints of the apical cell surfaces of thyrocytes that had surrounded the colloid globules in situ. Hence, in size and surface morphology, isolated colloid globules represent authentic lumenal content. Based on the total protein of single colloid globules and their volume, an average protein concentration of 590 mg/mL was calculated. The presence of protein disulfide isomerase in colloid globules and in the secretory product of cultured thyrocytes suggests its involvement in the extracellular multimerization of human TG. Native colloid globules increased their volume considerably upon reduction of disulfide bonds; they were completely dissolved by treatment with dithiothreitol and SDS. The results show that part of extracellular human TG undergoes multimerization, primarily by the formation of intermolecular disulfide bonds, thus allowing the storage of TG at excessively high, previously unknown, concentrations.

In vitro-staining specificity of the antibody 5-D-4 for microglia but not for monocytes and macrophages indicates that microglia are a unique subgroup of the myelomonocytic lineage.

We have previously shown that (i) the ramified phenotype and (ii) the microglia-specific pattern of membrane currents are induced not only in microglia, but also in monocytes and macrophages if they are cultured in the presence of astrocytes. These findings indicated that microglia are not a separate type of cell of the myelomonocytic lineage, but are induced to take on their unique characteristics by astrocytes. Recently, it was discovered that the antibody 5-D-4 selectively stains ramified microglia in situ. We therefore studied the influence of astrocytes and other epithelial cells on the expression of the keratan sulfate epitope recognized by 5-D-4 in microglia and other myelomonocytic cells. Our findings show that this antigen is exclusively expressed in microglia only if they are induced to ramify by coculture with either astrocytes or epithelial cells. By contrast monocytes and macrophages, even if induced to take on the ramified phenotype do not stain positive with 5-D-4. These findings indicate (i) that 5-D-4 is a specific marker for ramified microglia in vitro, and (ii) that microglia are a separate class of myelomonocytic cells, distinct from monocytes and macrophages.

ATP and adenosine induce ramification of microglia in vitro.

Microglial cells in the healthy adult brain possess a characteristic ramified morphology with multiple branched processes, small somata and down-regulated inflammatory properties. In contrast, microglial cells isolated from new-born rat brain inevitably show a non-ramified amoeboid phenotype, which is observed in vivo after pathologic activation or during development. To identify factors that control microglial morphology we investigated the effects of purines alone or in combination with astrocyte-conditioned medium (ACM). Under optimized culture conditions postnatal rat microglial cells developed an amoeboid to ovoid phenotype. Addition of 0.6-1 mM ATP or adenosine induced the outgrowth of numerous processes after 2-3 days that could be observed also in the presence of ACM as previously reported. Culture in ACM plus ATP or adenosine yielded an optimized ramified phenotype. ATP or adenosine, but not ACM alone, also prevented the formation of a flat, amoeboid morphology induced by lipopolysaccharide (LPS); however, at 0.6-1 mM they did not reduce the initial LPS-induced activation of the transcription factor NF-kappaB. By using specific agonists or antagonists the morphological transformations could not be confined to a distinct purinoreceptor subtype, but appeared to be mediated by long-term presence of adenosine in the medium to which phosphorylated purines were rapidly hydrolyzed by microglial cells. Since ACM did not contain sufficient concentrations of ATP or adenosine, purines are not the only ramification-inducing factors present in ACM; however, they are a valuable tool to induce microglial ramification in vitro.

Intrathecal synthesis of monocyte chemoattractant protein-1 (MCP-1) in amyotrophic lateral sclerosis: further evidence for microglial activation in neurodegeneration.

Autopsy studies and animal experiments suggest that microglial inflammation contributes to the pathogenesis of amyotrophic lateral sclerosis (ALS). Monocyte-chemoattractant protein (MCP-1) might play an important role in microglial recruitment. We studied MCP-1 levels in sera and cerebrospinal fluid of 29 ALS patients and compared the results with 11 control patients with tension headache. The MCP-1 level was determined using enzyme-linked immunosorbent assays (ELISA). A significant increase in cerebrospinal fluid MCP-1 level but not serum level was seen in the patients with ALS compared to the control subjects. These results suggest that cerebrospinal fluid MCP-1 activity may be a sensitive marker for neuroinflammation in ALS useful for monitoring treatment trials in ALS.

Reduction of degranulation of polymorphonuclear leukocytes by immunosuppression in patients following cadaveric renal transplantation.

Plasma levels of main granulocyte components of patients after cadaveric renal transplantation were compared 9 days postoperative with the plasma levels of patients undergoing aortofemoral and iliacofemoral bypass operation or abdominal surgery. Lactoferrin values were significantly lower in patients under immunosuppression with cyclosporine and prednisolone, whereas plasma levels of myeloperoxidase were comparable in all 3 groups of patients. Plasma E-alpha 1 PI values were significantly lower in patients undergoing bypass operation compared to abdominal surgery but did not differ from patients undergoing cadaveric kidney transplantation. Within 22 days postoperatively, there was no difference in the plasma levels of main granulocyte components in patients after kidney transplantation with and without postoperative complications. In vitro incubation of heparinized whole blood and isolated granulocytes obtained from healthy subjects in the presence of CsA, azathioprine, or prednisolone were performed. Only CsA caused inhibition of spontaneous degranulation of polymorphonuclear neutrophils showing significant lower elastase and lactoferrin release. However, in vivo administration of CsA and prednisolone in transplant patients displayed no effect on in vitro degranulation of both whole blood samples and isolated granulocytes. Our data demonstrate that CsA after in vitro incubation inhibits spontaneous granulocyte degranulation but not after in vivo administration. However, in vivo administration of CsA and prednisolone reduces lactoferrin release under certain conditions, e.g., postoperative stress or during hemodialysis therapy.

From theory to therapy: implications from an in vitro model of ramified microglia.

Microglia are the principal immune cells in the central nervous system (CNS), characterized by a highly specific morphology and unusual antigenic phenotype. An increasing number of studies have focused on the role of microglia in the pathogenesis of neurodegenerative diseases. To elucidate the function of microglial cells under several neuropathological conditions, we have studied and established a cell culture model that allows us to cultivate microglial cells in their inactive, resting (ramified) phenotype. In the first part of this work, we describe the interaction of microglia cells with their epithelial (astrocytic) microenvironment. The second part reviews experiments with microglia cell cultures to elucidate underlying signalling pathways and summarizes recent advances of our knowledge in microglial molecular pathways that may ultimately lead to neurodegeneration.

The pathophysiology of parkinsonian tremor: a review.

Parkinsonian tremor is most likely due to oscillating neuronal activity within the CNS. Summarizing all the available evidence, peripheral factors only play a minor role in the generation, maintenance and modulation of PD tremor. Recent studies have shown that not a single but multiple oscillators are responsible. The most likely candidate producing these oscillations is the basal ganglia loop and its topographic organization might be responsible for the separation into different oscillators which, nevertheless, usually produce the same frequency. The neuronal mechanisms underlying these oscillations are not yet clear, but three hypotheses would be compatible with the presently available data from animal models and data recorded in patients. The first is a cortico-subthalamo-pallido-thalamic loop, the second is a pacemaker consisting of the external pallidum and the subthalamic nucleus, and the third is abnormal synchronization due to unknown mechanisms within the whole striato-pallido-thalamic pathway leading to a loss of segregation. Assuming the oscillator within the basal ganglia pathway, the mechanism of stereotactic surgery might be a desynchronization of the activity of the basal ganglia-thalamo-cortical or the cerebello-thalamo-cortical pathway.

Cerebrospinal fluid from patients with neurodegenerative and neuroinflammatory diseases: no evidence for rat glial activation in vitro.

To determine the possible contribution of glial cells via oxidative stress/cytokine secretion in the pathogenesis of Parkinson's disease (PD), Alzheimer disease (AD), amyotrophic lateral sclerosis (ALS) or multiple sclerosis (MS) the concentration of nitric oxide (NO) (by the Griess method) and Interleukin-6 (IL-6) (by enzyme-linked immunosorbent assay) were measured in resting rat microglial and astrocytic cell culture supernatants stimulated by cerebrospinal fluid (CSF) (dilution 1:4, 1:10) from patients with the aforementioned diseases. Neither the concentration of NO (optical density at 450 nm: control, 0.036+/-0.006; MS, 0.034+/-0.008; AD, 0.031+/-0.006; PD, 0.02+/-0.01; lipopolysaccharide (LPS), 0.26+/-0.018) nor the amount of IL-6 (ng/ml: control, 0.112+/-0.026; PD, 0.12+/-0.027; MS, 0.123+/-0.008; ALS, 0.137+/-0.01; LPS, 1.81+/-0.11) differed in any disease group from those of unaffected controls. These findings suggest that the stimuli for inflammatory activation of glia are quite localized and not present in sufficient concentrations in the CSF of affected patients.

Effect of plasma protein binding, volume of distribution and molecular weight on the fraction of drugs eliminated by hemodialysis.

The correlation between plasma protein binding, the volume of distribution, molecular weight, and percentage removed by hemodialysis was investigated in 89 drugs using information available in the literature. The correlation was significantly linear between dialyzability and plasma protein binding, as well as with the reciprocal volume of distribution. This is in agreement with the theoretical deduction of dialyzability from diffusion and convection kinetics. Multiple linear regression analysis revealed that only 27% of the variance in dialyzability could be explained by plasma protein binding (17%), the volume of distribution (6%), and the molecular weight (4%) of the drugs. Therefore, the dialyzability of drugs can not be predicted reliably.

Daily life in very old age: everyday activities as expression of successful living.

The goals of this article are (a) to describe the daily life of the very old in terms of frequency, duration, variety, and social and physical contexts of activities, and (b) to examine the effects of background variables (e.g., age, sex, residential and marital status, income, and education) on late life activity engagement. A representative sample of 516 adults aged 70-105 was interviewed about their activities using the Yesterday Interview. In contrast to most research on activity engagement, this measurement approach allows for assessment of both the type and context of activities engaged in during the day preceding the interview. The results indicated high frequencies of obligatory activities but also showed substantial time spent in discretionary activities, with television viewing occupying most of the participants' leisure time. Most activities were done alone and at home. In bivariate and multiple regression analyses, age and residential status had the strongest association with activity frequency, duration, and variety; the oldest-old and those residing in long-term care facilities had lower levels of activity engagement. Results are discussed in terms of their relevance for successful aging.

[Function study of the peritoneovenous shunt system]. / Die Funktionsuntersuchung des peritoneovenösen Shuntsystems.

In patients with otherwise therapy-resistant ascites operative methods for reinstillation of ascitic fluid have gained importance in recent years. Most often peritoneovenous shunt systems in the modification "LeVeen" or "Denver" are set in. Clinical evidence for insufficiency of the shunt system can force the radiologist to check the function of the drainage system. The investigation has to be performed under sterile conditions on the fluoroscope table, thus permitting the angiographic assessment of thrombotic complications as well as check of the position and function of the shunt valve and drainage tubes.

Successful transplantation of adult and fetal mouse pancreas islet cells using anti-class II antibodies.

Fetal pancreas islet tissue of mice is able to proliferate in cell culture. The insulin production after 14 days is sufficient to reverse diabetes mellitus. The immunogenicity is reduced by tissue culture. By using monoclonal antibodies it could be shown that the expression of class II antigens on the surface of the cells is lower after tissue culture than before. Together with complement, the positive cells could be lysed, and transplantation even in completely different mice stems was successful.

[Cranial complications in extension therapy of cervical fractures. Riechert's modification of extension therapy]. / Kranielle Komplikationen bei der Extensionsbehandlung von Frakturen der Halswirbelsäule. Riechert'sche Modifikation der Extensionsbehandlung.

Cranial complications are reported in the treatment of cervical injury by Crutchfield skull caliper traction. The mechanical causes are discussed and a new skull caliper is demonstrated.

[Management of ureteral stenoses and urinary fistulas following kidney transplantation]. / Die Behandlung von Ureterstenosen und Urinfisteln nach Nierentransplantationen.

The treatment of strictures and fistulas at the lower ureter after kidney transplantation was simple and could be achieved by reimplantation into the bladder. Proximal urinary fistulas caused early abdominal symptoms. The i.v. urogram showed a dilated renal pelvis without drainage into the ureter. The anatomical findings were in all cases strictures or total obstruction of the ureter beneath the pelvic junction and a rupture of the renal pelvis or calix. Adequate therapy consisted of ureteroureterostomy with the recipient ureter and nephrostomy splintage.

[Treatment of renal artery stenosis following kidney transplantation]. / Therapie der Nierenarterienstenose nach Nierentransplantation.

Stenoses of the renal artery occurred in 4.6% of 415 transplanted kidneys between 1969 and 1983. Most of them were caused by traumatic intima-lesions and were localised postanastomotically. The best therapeutic procedure seems to be the transperitoneal approach with widening venous patch plasty or venous bypass. The percutaneous transluminal catheter dilatation may be successful in suitable cases.

[Sclerosing peritonitis following continuous ambulatory peritoneal dialysis]. / Die sklerosierende Peritonitis nach kontinuierlicher ambulanter Peritoneal-Dialyse (CAPD).

Sclerosing peritonitis is a severe complication after CAPD treatment. The visceral peritoneum is thickened and interenteric adhesive parts are found. Myofibroblasts are proliferated and the collageneous tissue is hyperplastic. The mean clinical symptom is the mechanical obstruction of the small bowel. We observed this illness in three out of sixty patients under CAPD. These patients had higher incidence of bacterial peritonitis. In the ascites high concentrations of PG E2 and Thromboxan B2 were observed. After treatment of the infection the concentrations fell down to normal values. Electronoptical observations from peritoneal biopsies showed a proliferation of myofibroblasts and extracellular lysosomes. It is known from these lysosomes that they are able to set free proteasis. These lead to degredation of fibrin and fibrinogen. These splits are mitogen to myofibroblasts. release from HIT cells could also be evoked by the sulphonylureas glibenclamide and tolbutamide and by an increase in concentration of extracellular K+ to 40 mmol/l. The content of cyclic AMP in HIT cells was increased modestly by glucose but not by an increase in extracellular K+. Forskolin elicited a 4-fold increase in cyclic AMP content. We conclude that HIT cells retain the essential features of the insulin secretory response of normal B cells and represent an important tool for further biochemical characterisation of the secretory system.