Employment status 1 year after out-of-hospital cardiac arrest in comatose patients treated with therapeutic hypothermia.

BACKGROUND: Therapeutic hypothermia for comatose survivors of out-of-hospital cardiac arrest (OHCA) has improved survival and neurologic outcome. This study focused on return to work 1 year after therapeutic hypothermia. METHODS: From June 2004 to June 2009, patients between 18 and 65 years of age with OHCA, who were treated with hypothermia from two regions, representing one third of the national population, were identified from the Danish National Patient Registry, and from hospital and ambulance records. The patients' employment status was obtained from the Danish Ministry of Employment. RESULTS: One hundred thirty-three comatose patients after OHCA treated with hypothermia were identified. One hundred and four (78%) patients were employed, or able to work, at the time of cardiac arrest. This particular group of patients showed significant lower in-hospital mortality compared to the group of patients who were not able to work before cardiac arrest; 13% vs. 48%, respectively (P < 0.001). The workable group had a lower Charlson comorbidity score (P = 0.004), a higher incidence of witnessed cardiac arrest (P = 0.004) and a higher incidence of shockable heart rhythm (P < 0.001). Eighty-seven patients (84%), who were able to work prior to cardiac arrest, survived, and 55 (65%) of these patients were employed or able to work at 1 year follow-up. CONCLUSION: The majority of patients employed, or able to work prior to OHCA, had returned to work at one year follow-up. Predictors of return to work in comatose patients treated with hypothermia have to be identified in a larger-scale study.

Synchrotron vacuum ultraviolet radiation studies of the D (1)Π(u) state of H(2).

The 3pπD (1)Π(u) state of the H(2) molecule was reinvestigated with different techniques at two synchrotron installations. The Fourier transform spectrometer in the vacuum ultraviolet wavelength range of the DESIRS beamline at the SOLEIL synchrotron was used for recording absorption spectra of the D (1)Π(u) state at high resolution and high absolute accuracy, limited only by the Doppler contribution at 100 K. From these measurements, line positions were extracted, in particular, for the narrow resonances involving (1)Π(u) (-) states, with an accuracy estimated at 0.06 cm(-1). The new data also closely match multichannel quantum defect calculations performed for the Π(-) components observed via the narrow Q-lines. The Λ-doubling in the D (1)Π(u) state was determined up to v=17. The 10 m normal incidence scanning monochromator at the beamline U125/2 of the BESSY II synchrotron, combined with a home-built target chamber and equipped with a variety of detectors, was used to unravel information on ionization, dissociation, and intramolecular fluorescence decay for the D (1)Π(u) vibrational series. The combined results yield accurate information on the characteristic Beutler-Fano profiles associated with the strongly predissociated Π(u) (+) parity components of the D (1)Π(u) levels. Values for the parameters describing the predissociation width as well as the Fano-q line shape parameters for the J=1 and J=2 rotational states were determined for the sequence of vibrational quantum numbers up to v=17.

Massive upper GI bleeding: a rare complication of Zenker's diverticulum.

Bleeding from a Zenker's diverticulum is rare. A 71-year-old man was urgently admitted with massive hematemesis. It was known that he had a Zenker's diverticulum, but on emergency endoscopy, the source of bleeding was not detected due to large blood clots in the esophagus, hypo-pharynx and also into the tracheal-bronchial tree. Computerized tomography angiography demonstrated a blush of intravenous contrast arising from the diverticulum. The patient was operated upon urgently; the diverticle had a deep ulceration which was the source of the bleeding. The cause of the ulceration is unknown but it is possible that it was caused by the direct effect of an aspirin pill within the diverticle. A similar case with the same conclusion has been published in the past and since the use of aspirin has become common, especially in the elder population, we present this case report to highlight this possible life-threatening complication of Zenker's diverticulum in patients receiving aspirin.

Dissociation from BiP and retrotranslocation of unassembled immunoglobulin light chains are tightly coupled to proteasome activity.

Unassembled immunoglobulin light chains expressed by the mouse plasmacytoma cell line NS1 (kappa(NS1)) are degraded in vivo with a half-life of 50-60 min in a way that closely resembles endoplasmic reticulum (ER)-associated degradation (). Here we show that the peptide aldehydes MG132 and PS1 and the specific proteasome inhibitor lactacystin effectively increased the half-life of kappa(NS1), arguing for a proteasome-mediated degradation pathway. Subcellular fractionation and protease protection assays have indicated an ER localization of kappa(NS1) upon proteasome inhibition. This was independently confirmed by the analysis of the folding state of kappa(NS1) and size fractionation experiments showing that the immunoglobulin light chain remained bound to the ER chaperone BiP when the activity of the proteasome was blocked. Moreover, kinetic studies performed in lactacystin-treated cells revealed a time-dependent increase in the physical stability of the BiP-kappa(NS1) complex, suggesting that additional proteins are present in the older complex. Together, our data support a model for ER-associated degradation in which both the release of a soluble nonglycosylated protein from BiP and its retrotranslocation out of the ER are tightly coupled with proteasome activity.

Protein-mediated protein maturation in eukaryotes.

Eukaryotic cells have developed particular strategies to support the critical steps in protein maturation that starts in the cytosol with the birth of a nascent polypeptide chain, and ends when the protein has reached the appropriate compartment and/or has attained its mature structure. Many of the cellular proteins that have evolved to promote maturation processes are constitutively expressed members of the highly conserved heat shock protein (hsp) family, also known as 'molecular chaperones'. Protein-mediated processes that occur in the cytosol are discussed.

Expression of highly active sex-inducing pheromone of Volvox carteri f. nagariensis in a mammalian cell system.

A cDNA fragment coding for the sex-inducing glycoprotein of Volvox carteri f. nagariensis was expressed in a mammalian cell system (baby hamster kidney (BHK) cells). The transfection product exhibited a specific biological activity intermediate between the natural pheromone of the strains Volvox carteri f. nagariensis and Volvox carteri f. weismannia. Immunoblot analysis showed that the sex-inducing activity was expressed as a set of three iso-glycoproteins (35, 34 and 31 kDa).

Effect of the aggregational state on the mitogenicity of lipoprotein from the outer membrane of Escherichia coli.

The effects of ultrasonication, electrodialysis, and centrifugation through sucrose gradients on the mitogenicity of the lipoprotein (LP) from the outer membrane of Escherichia coli were studied in several inbred mouse strains. LP is not readily soluble in aqueous solution. By ultrasonication for 1 minute we obtained LP aggregates which were fully mitogenic; prolonged sonification had no influence on activity. Solubilization in 4% sodium dodecyl sulphate (SDS) and centrifugation into sucrose gradients yielded different aggregation forms of LP, which were B-lymphocyte mitogens towards LPS non responder C3H/HeJ mouse splenocytes, and towards congenitally athymic nude mice. By electrodialysis, LP could be transformed into different salt forms. The TEA-salt of LP exhibited enhanced mitogenicity. The results suggest that the mitogenicity of lipoprotein is widely independent from the aggregational state in which it is offered to the cells.

Antitumor activity of anti-epidermal growth factor receptor monoclonal antibodies and cisplatin in ten human head and neck squamous cell carcinoma lines.

Head and neck squamous cell carcinomas (HNSCC) frequently display increased levels of epidermal growth factor receptor (EGFR) and since the receptor is located on the cell surface, anti-EGFR antibodies appear to be suitable agents for antitumor therapy. We investigated the effect of murine EMD 55900 and rat ICR 62 monoclonal antibodies (MAb) directed against EGFR both as single agents and in combination with cisplatin. ELISA detection showed the amount of EGFR protein in HNSCC lines UM-SCC-10A, -10B, -11B, -14A, -14B, 14C, -22B and HLac 79, 8029NA, 8029DDP to range between 20 and 8100 fmol/mg protein. Compared to A431 cells, seven HNSCC lines were high and three low receptor expressors. Only low levels of TGF alpha were found in the supernatants of some untreated HNSCC lines, probably due to the consumption of TGF alpha by EGFR. Consequently, occupation of EGFR by MAb led to marked accumulation of TGF alpha in cell supernatants. Colorimetric MTT assay showed both MAbs (0.3-30nM) to have comparable dose-dependent growth inhibition which correlated with the EGFR content of the respective cell lines (p < 0.05). Using 30nM MAb, seven high receptor expressing HNSCC lines were growth inhibited by at least 20% to a maximum of 61% (mean = 38%). Combined treatment with MAb and cisplatin led to a significant decrease in cisplatin IC50 values in 5 cell lines expressing more than 1200 fmol EGFR/mg (dose modification by factor 2.1-4.1). In conclusion, anti-EGFR MAb exert direct antiproliferative activity in HNSCC lines and show additive effects in combination with cisplatin.

Short-term results and long-term patients' appraisal of abdominal colposacropexy for treatment of genital and vaginal vault prolapse.

OBJECTIVE: To evaluate short-term results and long-term patients' satisfaction of abdominal colposacropexy performed for massive genital prolapse in our institute during a 14-year period. STUDY DESIGN: A retrospective analysis was performed of 101 consecutive women who underwent abdominal colposacropexy. The procedure consisted of retroperitoneal interposition of a Mersilene mesh between a prolapsed vaginal vault or uterus and the anterior surface of the sacrum. A questionnaire was used to evaluate patients' satisfaction after surgery. RESULTS: In the short-term there was an acceptable rate of complications of surgery, comparable with that reported in the literature. Prolapse-related complaints evaluated by questionnaire were markedly diminished after surgery. Pain and functional complaints were only partly improved in a subgroup of patients. Only 32% of the patients responded that they were fully cured after operation. CONCLUSIONS: Abdominal colposacropexy shows a particularly favorable result on prolapse-related complaints. Functional complaints and pain are not substantially relieved by this procedure. The complication rate of surgery is acceptable.

Dual role of fibroblasts in tumor cell growth.

Vast experience with cultivating biopsies from human tumors indicates that in most cases the admixture of fibroblasts has a negative effect on growth of tumor cells. Only rarely is observed help provided by the fibroblasts. It has also long been known that fibroblasts can inhibit by contact themselves and also produce growth factor(s) promoting cell proliferation. We have used three permanent squamous cell carcinoma lines and fibroblasts derived from biopsies of trachea to study this paradox. The inhibitory activity of fibroblasts is contact-dependent in a cell membrane-bound factor, which is trypsin sensitive. We prepared microsomal fractions (MF) from aged (more than 40 passages) fibroblasts and followed their effect on proliferation of the tumor cell lines in MTT assay. MF from all three fibroblast lines inhibited the tumor cells. Most regularly was this phenomenon observed with line UM-SCC 22B. Not all tumor cells are immortal. On the contrary, a large portion of them undergoes terminal differentiation. With the line UM-SCC 22B we tested the possibility that MF from fibroblasts can increase the portion of tumor cells which will senescence after few mitoses. The immortal cells were defined as cells capable in 8 or 13 days of forming colonies of more than 50 or 200 cells. The senescent cells were defined as cells not capable of producing within the same period of time colonies of more than 12 or 30 cells. The MF was able to increase the number of small colonies, i.e. the number of senescent tumor cells. The fibroblasts of the same passage level were releasing soluble growth factor(s) promoting growth of cells. Fibroblasts can apparently simultaneously inhibit growth by contact and release factor(s) promoting growth of cells. The final outcome is a result of the balance between these two forces. This balance is regulated by many intrinsic and extrinsic forces.

[Prevention of neonatal tetanus in developing countries hampered by local organization and limited knowledge of health personnel and traditional midwives; North Sulawesi (Indonesia)]. / Preventie van neonatale tetanus in ontwikkelingslanden bemoeilijkt door lokale organisatie en beperkte kennis bij gezondheidswerkers en traditionele vroedvrouwen; Noord-Sulawesi (Indonesië).

OBJECTIVE: Evaluation of the policy to prevent neonatal tetanus in North-Sulawesi (Indonesia). DESIGN: Descriptive and questionnaire study. SETTING: Twelve villages in the district of Minahassa, Sulawesi, Indonesia. METHOD: The policy aimed at preventing neonatal tetanus was studied during the period April-June 1992. The relevant knowledge was tested of the community health workers (CHWs, n = 142) and traditional birth attendants (TBAs, n = 37) active in the 12 villages. To this purpose, 54 and 27 workers, respectively, were interviewed using a standard questionnaire. RESULTS: Not all women were vaccinated sufficiently to prevent their unborn children from tetanus; mostly, adequate vaccination records were lacking. 33% of the TBAs were unaware that neonatal tetanus can be prevented by hygienic delivery; in this respect there was no difference between trained and untrained TBAs. Of the CHWs, 39% were familiar with the correct vaccination policy and 26% did not know that pregnant women have to be vaccinated. 70% of the CHWs and TBAs wrongly believed that neonatal tetanus no longer occurred in their village. CONCLUSION: CHWs and TBAs showed little interest in neonatal tetanus, so that they were lacking in motivation regarding hygienic delivery procedures and preventive tetanus toxoid vaccination. Implementation of a good preventive programme in developing countries mostly requires organizatory rather than biomedical improvement.

The CIMT-monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8+ T lymphocytes by structural and functional assays.

The interpretation of the results obtained from immunomonitoring of clinical trials is a difficult task due to the variety of methods and protocols available to detect vaccine-specific T-cell responses. This heterogeneity as well as the lack of standards has led to significant scepticism towards published results. In February 2005, a working group was therefore founded under the aegis of the Association for Immunotherapy of Cancer ("CIMT") in order to compare techniques and protocols applied for the enumeration of antigen-specific T-cell responses. Here we present the results from two consecutive phases of an international inter-laboratory testing project referred to as the "CIMT monitoring panel". A total of 13 centers from six European countries participated in the study in which pre-tested PBMC samples, synthetic peptides and PE-conjugated HLA-tetramers were prepared centrally and distributed to participants. All were asked to determine the number of antigen-specific T-cells in each sample using tetramer staining and one functional assay. The results of the first testing round revealed that the total number of cells analyzed was the most important determinant for the sensitive detection of antigen-specific CD8(+) T-cells by tetramer staining. Analysis by ELISPOT was influenced by a combination of cell number and a resting phase after thawing of peripheral blood mononuclear cells. Therefore, the experiments were repeated in a second phase but now the participants were asked to change their protocols according to the new guidelines distilled from the results of the first phase. The recommendations improved the number of antigen-specific T-cell responses that were detected and decreased the variability between the laboratories. We conclude that a two-step approach in inter-laboratory testing allows the identification of distinct variables that influence the sensitivity of different T-cell assays and to formally show that a defined correction to the protocols successfully increases the sensitivity and reduces the inter-center variability. Such "two-step" inter-laboratory projects could define rational bases for accepted international guidelines and thereby lead to the harmonization of the techniques used for immune monitoring.

BiP (GRP78), an essential hsp70 resident protein in the endoplasmic reticulum.

BiP is a constitutively-expressed resident protein of the endoplasmic reticulum (ER) of all eucaryotic cells, and belongs to the highly conserved hsp70 protein family. In the ER, BiP is involved in polypeptide translocation, protein folding and presumably protein degradation as well. These functions are essential to cell viability, as has been shown for yeast. In this review, I will summarize the structural features of hsp70 proteins and focus on those experiments which revealed the biological function of BiP.

Does sophisticated diagnostic workup on neuroectodermal tumors have an impact on the treatment of esthesioneuroblastoma?

BACKGROUND: The diagnostic workup on esthesioneuroblastoma is more extensive than ever before. We have investigated whether improvements in diagnosis of sinonasal neuroectodermal tumors, including esthesioneuroblastomas (ENB), sinonasal neuroendocrine carcinomas (SNEC) and sinonasal undifferentiated carcinomas (SNUC), have had an impact on treatment and outcome. PATIENTS AND METHODS: 11 ENB, 7 SNEC and 1 SNUC in 13 men and 6 women (average age 52.9 years (range 26-82)), diagnosed between 1986 and 2001, were analyzed with regard to histopathologic and clinical diagnosis as well as outcome. Our results were compared with the available literature. RESULTS: According to the Morita classification considering endoscopy, CT and MRI scans, 2 tumors were staged D, 14 were found to be stage C, 2 were stage B and 1 was stage A. Lightmicroscopically only 4 of 19 showed higher differentiation and rosette-like structures, the others were poorly differentiated. 18 of 19 tumors were examined immunohistochemically. Neuronal markers (NSE, synaptophysin, chromogranin, S-100 and neurofilaments) were heterogeneously expressed in both ENB and NEC, only NSE stained all but 2 tumors. Coexpression of neuronal markers and cytokeratins was proven in all NEC and 5 of 11 ENB. Some tumors expressed atypical markers. Despite extensive diagnostic steps it was not possible to exclude a different histopathological diagnosis in 10 of 19 cases. CONCLUSION: For sinonasal neuroectodermal tumors no pathognomonic antigenic profiles are known. Immunohistochemical markers lack specificity and sensitivity. Nevertheless, in many sinonasal neuroectodermal tumors a panel of differentiation markers allows to specify the light-microscopic diagnosis. Until now no therapeutic consequence arises from a more extensive diagnostic workup. However, the histopathologic identification of subtypes (SNUC) and proliferation markers may help to identify patients with poor prognosis.

[Expression of cellular antigen of hypopharyngeal carcinoma in different culture conditions]. / Expression von zellulären Antigenen eines Hypopharynxkarzinoms unter verschiedenen Kulturbedingungen.

BACKGROUND: The need to improve therapy regimes, determine prognosis, and study biological properties of tumors extracorporally led to development of different experimental systems. In order to approach the in vivo situation, specific properties of the tumors of origin should be retained by the cells in culture over relatively long periods. However, culture conditions may change expression of cellular antigens. METHODS: Cryosections of a hypopharyngeal carcinoma were compared in this respect with different cultivation systems (2-dimensional monolayers [ML], 3-dimensional multicellular tumor spheroids [MTS] and substrate cultures on Gelita) in regard to expression of cytokeratins (CK) 1, 7, 10, 14, 18 and 19, vimentin, neurofilament (NF) kD200 and 68, ganglioside GD2, oncogene products (P53 mutant and wild), and membrane-associated antigens (HLA-ABC and -DR, epidermal growth factor receptor EGFR). RESULTS: Semiquantitative immunohistochemical methods revealed differences in expression of CK1, 14 and 19, GD2, and P53 mutant between these systems. CONCLUSION: Pronounced expression of markers in MTS compared to original biopsy and monolayer emphasizes the importance of cell-cell contact and 3-dimensionality or metabolic stress. However, weak marker expression within substrate cultures may reflect loose cell-cell contact observed. In these experiments, the antigenic configuration of MTS resembled the one of the original tumor more than the other culture systems: monolayer and growth on substrate. As vimentin and NF are not expressed by healthy epithelial cells of adults, occurrence of intratumoral vimentin and NF could point to derepression of early differentiation antigens.

Immunoglobulin heavy chain toxicity in plasma cells is neutralized by fusion to pre-B cells.

A plasma cell hybridoma frequently loses its immunoglobulin heavy (H) chain spontaneously but rarely is production of its light (L) chain lost. Upon fusion to a pre-B-cell hybridoma that produces no Ig chain, the L chain is frequently lost. In cells without the L chain the H chain, which is derived from the plasma cell, is not chemically modified. Our results indicate that, in pre-B cells, but not in plasma cells, there must be a mechanism that neutralizes the toxic effect of free H chain.

Interaction of BiP with newly synthesized immunoglobulin light chain molecules: cycles of sequential binding and release.

Here we show that not only transport defective but all immunoglobulin light chains interact with BiP. Association of BiP with its ligand takes place during or shortly after translation of the light chains. The biological half life of the BiP-light chain complex depends on the fate of the light chains. Light chains which are secreted interact with BiP for only a very short time. In contrast, the complex is biologically more stable in cells which do not secrete their L chains. In these cells, dissociation from BiP correlates with the biological half life of the L chains arguing for a degradation pathway in the endoplasmic reticulum. Instead of being degraded in association with its ligand, BiP is released from the complex and binds to newly synthesized polypeptides. These results support the notion that both H and L chains require the chaperoning function of BiP before or during the process of antibody assembly.