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Biomedical subjects

R Jakob

Publications and source records attributed to R Jakob.

At least 19 recordsLinked to original sources

[Aortoduodenal fistula as a cause of gastrointestinal bleeding--difficulties in endoscopic diagnosis].

We report on three patients with severe gastrointestinal bleeding arising from aortoenteric fistula. Two patients presented with a secondary aortoduodenal fistula. In the first case bleeding occurred 8 months after aortobifemoral graft implantation. In the second patient aortobiliacal graft implantation was performed 22 years before. In the third case the aortoenteric fistula was primary and was caused by an abdominal aortic aneurysm without prior vascular intervention. In the first case diagnosis was made by urgent endoscopy visualizing ongoing bleeding from the duodenal fistula. In the two other patients urgent endoscopy and CT as well could not demonstrate the bleeding source. Aortoenteric fistula was diagnosed endoscopically during severe rebleeding some hours later. Two patients underwent surgery with implantation of an axillobifemoral bypass; the third patient declined further intervention and died. The course shows that aortoduodenal fistula can present with severe but intermittent gastrointestinal bleeding making the diagnosis in the non-bleeding interval difficult. In patients with severe gastrointestinal bleeding and a history of aortic disease (aneurysm, prior aortic graft repair or stenting) an aortoduodenal fistula should be suspected and the indication for surgical intervention should be considered early in spite of negative results of endoscopy and CT.

Aged↗

Increased PGE2 levels in nonmalignant mucosa adjacent to squamous cell carcinoma of the head and neck.

OBJECTIVE: The cyclooxygenases (COX) 1 and 2 are the rate-limiting enzymes of prostaglandin E(2) (PGE(2)) synthesis, and the upregulation of COX-2 has been reported in tumors of different origins. The aim of our study was to quantify the PGE(2) expression in squamous cell carcinoma and surrounding mucosa, to analyze the potential of acetylsalicylic acid (ASA) for reducing PGE(2) levels in these tissues, and to improve our understanding of potential tumor-derived stimulation of surrounding mucosa by PGE(2). STUDY DESIGN AND SETTING: Intracellular PGE(2) levels in primary head and neck squamous cell carcinoma (HNSCC) and the surrounding mucosa at 1 and 2 cm distance were analyzed ex vivo by ELISA. Subsequently, we treated in vitro tumor and normal mucosal cells from turbinates with recombinant PGE(2) and ASA, and quantified intracellular PGE(2) levels. RESULTS: We observed high PGE(2) levels in the tumor samples and in tumor-surrounding mucosa. The addition of PGE(2) and arachidonic acid to tumor cell cultures resulted in no further increase in intracellular PGE(2) levels, while ASA reduced PGE(2) levels by up to 40%. In normal epithelial cell cultures, less PGE(2) (6% of that found in the tumor cells) was expressed, but stimulation with PGE(2) resulted in levels comparable to those of the tumor samples. CONCLUSION AND SIGNIFICANCE: We conclude that HNSCC and the surrounding mucosa express high levels of PGE(2). This expression is reduced efficiently by ASA. We propose a stimulation of PGE(2) expression in the epithelium surrounding HNSCC by tumor-derived PGE(2) with a potential impact on tumor growth.

Carcinoma, Squamous Cell↗

Relevance of primary reduction for the clinical outcome of femoral neck fractures treated with cancellous screws.

INTRODUCTION: Femoral head-preserving treatment for femoral neck fractures with cancellous screws is a method widely performed, but its success depends on the quality of primary reduction. The goal of our study was to evaluate the relevance of reduction quality on the clinical outcome. PATIENTS AND METHODS: We retrospectively analyzed the postoperative reduction result of 39 patients. The angular rotation and the inferior displacement of the proximal fracture fragment were digitally assessed from AP radiographs. The clinical outcome was recorded by a follow-up after a mean 5.5 years. RESULTS: Unsuccessful healing was observed in eight patients, femoral head necrosis in three patients, non-union in two patients, implant loosening in one patient, and posttraumatic arthritis in two patients, all resulting in the need for a total hip replacement. This clinical course was strongly associated with a primary reduction in varus position, whereas reduction in anatomic or valgus angulation had no negative effect on the clinical outcome. An inferior offset was no negative predictor for successful healing. CONCLUSION: We conclude that a non-anatomic reduction in varus angulation significantly increases the risk of healing failure and therefore should be avoided. Reduction in valgus position should be performed if secondary redislocation is imminent.

Adult↗

How should we treat malignant pleural mesothelioma (MPM)?

MPM is an occupational disease with increasing incidence. From 1986 to 1998 we operated 59 patients for MPM: in 11 cases only biopsy or exploration, 23 (stage T1a and T1b) Pleurectomy/Decortication, 22 (stage T2/3) extended pleuropneumonectomy (PPE) with resection of pericardium and diaphragm. In 3 cases we did additionally a chestwall resection. One patient in the PPE-group died postoperatively. Median survival time in the diagnosis/exploration group is 6 months. In the pleurectomy group (earlier stages) 14 months, in the PPE-group 16 months. One patient in the PPE-group survived 68 months, in the pleurectomy group the longest survival was 39 months. Up today 20 patients in the PPE-group died, 12 without any evidence local recurrency. Despite the fact that patients in the pleurectomy group are operated in earlier tumor stages they do not have a longer median survival time. In more than 50 percent only local disease control can be achieved by ePPE. Therefore in my opinion the first operative step must be followed by a second therapeutic step as immune- or gene therapy.

Female↗

Influence of flupirtine on a G-protein coupled inwardly rectifying potassium current in hippocampal neurones.

1. Previous studies have shown that flupirtine, a centrally acting, non-opioid analgesic agent, also exhibits neuroprotective activity in focal cerebral ischaemia in mice and reduces apoptosis induced by NMDA, gp 120 of HIV, prior protein fragment or lead acetate as well as necrosis induced by glutamate or NMDA in cell culture. To study the potential mechanism of the neuroprotective action of flupirtine, we investigated whether flupirtine is able to modulate potassium or NMDA-induced currents in rat cultured hippocampal neurones by use of the whole-cell configuration of the patch-clamp technique. 2. We demonstrated that 1 microM flupirtine activated an inwardly rectifying potassium current (K(ir)) in hippocampal neurones (deltaI=-39+/-18 pA at -130 mV; n=10). This effect was dose-dependent (EC50=0.6 microM). The reversal potential for K(ir) was in agreement with the potassium equilibrium potential predicted from the Nernst equation showing that K(ir) was predominantly carried by K+. Furthermore, the induced current was blocked completely by Ba2+ (1 mM), an effect typical for K(ir). 3. The activation of K(ir) by flupirtine was largely prevented by pretreatment of the cells with pertussis toxin (PTX) indicating the involvement of a PTX-sensitive G-protein in the transduction mechanism (deltaI=-3+/-6 pA at -130 mV; n=8). Inclusion of cyclic AMP in the intracellular solution completely abolished the activation of K(ir) (n=7). 4. The selective alpha2-adrenoceptor antagonist SKF-86466 (10 microM), the selective 5-HT1A antagonist NAN 190 as well as the selective GABA(B) antagonist 2-hydroxysaclofen (10 microM) failed to block the flupirtine effect on the inward rectifier. 5. Flupirtine (1 microM) could not change the current induced by 50 microM NMDA. 6. These results show that in cultured hippocampal neurones flupirtine activates an inwardly rectifying potassium current and that a PTX-sensitive G-protein is involved in the transduction mechanism.

Aminopyridines↗

Electroporation-mediated delivery of nucleolar targeting sequences from Semliki Forest virus nucleocapsid protein.

Electroporation was used as a powerful and simple method to probe to the intracellular distribution and trafficking of signal sequences. By coupling synthetic peptides to carrier reporter groups, specific amino acid sequences responsible for nucleolar targeting of Semliki Forest virus (SFV) Core (C) protein were found out. In the N-terminal part of the C protein the sequences 66KPKKKKTTKPKPKTQPKK83 and 92KKKDKQADKKKKP105 are able to situate BSA or KLH as reporter proteins in the nucleolus, suggesting that SFV C protein contains at least two independent nucleolar targeting sequences.

Amino Acid Sequence↗

Six standard panels for hepatoviral diagnosis.

From the many available tests for hepatoviral diagnosis the most reliable and established were incorporated into a series of panels. Six groups or batteries of tests are proposed, each directed to one clinical question. These are hepatitis B detection and chronic hepatitis B monitoring, hepatitis detection and chronic hepatitis monitoring, as well as a hepatitis and chronic hepatitis monitoring, as well as a hepatitis screening panel for blood banks and an immune status panel to determine individual immune status and risk. This approach limits the number of immuno-diagnostic tests needed and simplifies laboratory workflow. The limitations, including specificity problems encountered with tests based on the polymerase chain reaction (PCR), and probable future developments of tests excluded from the simplified panel concept are discussed.

Hepatitis, Viral, Human↗

Nucleolar accumulation of Semliki Forest virus nucleocapsid C protein: influence of metabolic status, cytoskeleton and receptors.

The nucleolar accumulation of Semliki Forest Virus (SFV) C protein was examined as a function of intact microtubules, intact microfilaments and accessible intermediate filaments. The cytoskeletal components do not seem to play a role in directing C protein to the nucleolus but nucleolar accumulation is energy-dependent and saturable. This suggests the involvement of some receptor- (or chaperon-) interaction.

Animals↗

Nucleolar accumulation of core protein in cells naturally infected with Semliki Forest virus. Quantitative aspects.

The core (C) protein of Semliki Forest virus (SFV) is known to exert several important functions with regard to both the virus and the host. This paper shows that migration of parental and progeny C protein to the nucleolus is a common feature in infected vertebrate and invertebrate cells. The amount of C protein accumulating to the nucleolus is small, always less than 1% of the intra- and extracellular C protein at various times post infection. Migration to the nucleolus is a fast process; 1.5 h post infection a prominent amount of parental C protein is already incorporated into nucleolar fractions.

Aedes↗

Pathogenesis of alphavirus infection as demonstrated by infection of ECV 304 transformed human umbilical vein capillary endothelial cells with Semliki Forest virus.

Routes of alphaviral entry have been studied with the transformed human endothelial vein cell line ECV 304. This cell line can be cultured on tissue culture inserts which permits apical (lumenal) and basolateral (ablumenal) cell surfaces to be infected and observed separately. Semliki Forest Virus (SFV), a prototype alphavirus, was able to infect and replicate from both the apical and basolateral sides. Transcytosis is not the route by which SFV passes the endothelial cell barrier as demonstrated by polarised infection of junctionally tight ECV monolayers in which translation was inhibited. A "grow-through" replication may play a role in SFV pathogenesis. Infected ECV cells produced interfering substances that inhibited viral infection. Higher multiplicities of infection resulted in infection and complete destruction of the monolayer.

Animals↗

A multiple centre clinical study of third-generation enzyme immunoassays for hepatitis B surface antigen and hepatitis B core IgM class antibody.

Two new monoclonal antibody-based automated enzyme immunoassays for hepatitis B markers were tested for their clinical performance. The assays specifically detect hepatitis B surface antigen (HBsAg) and hepatitis B core (HBc) IgM antibodies by radial partition fluorescence enzyme immunoassay. Taken together, the HBsAg and IgM anti-HBc assay show clinical sensitivities of 99.5% and 95.6% and respective clinical specificities of 99.8% and 99.5%, compared with standard ELISAs. The detection limit goes down to 50 PEI units per litre for HBsAg ad subtype and 70 PEI units per litre for ay subtype.

Antibody Specificity↗

Technique and results of vascular endoscopy in arterial and venous reconstructions.

In an effort to maximize results, vascular endoscopy was used in our institution to monitor arterial and venous reconstructions. Since 1982, angioscopy was applied as a control method in 182 venous thrombectomies to treat iliofemoral thrombosis and 114 aortoiliac thromboendarterectomies. Of the cases with venous thrombectomy reviewed, 50% were incomplete by endoscopic evidence; of these, in 80% the remaining clots could be partly or completely removed. Additionally, in six patients a venous spur was found. Of 114 attempted aortoiliac thromboendarterectomies, only 91 could be completed. In the remainder, endoscopic evidence of persistent intimal flaps forced us to bypass the affected segments. With further miniaturization of the angioscopes, the method was also applied to check vessel repair on small-caliber arteries. In an initial study with 220 femorodistal bypasses we were unable to find a statistically significant difference of primary patency in grafts that were endoscopically controlled or not. In the learning phase with the in situ technique, we identified competent valve remnants in 40%, but this rate could be reduced to 12.7% with growing experience in valvulotomy. We conclude from our data that angioscopy is very helpful in assessing the morphological integrity of aortoiliac thromboendarterectomies and venous thrombectomies. The actual value in infrainguinal arterial reconstructions still remains to be proven.

Angioscopy↗

Purifying nuclei.

Nuclear isolation methods exist since over 50 years and even today new procedures and amendments of standard methods are published. They can be classified into nonaqueous and aqueous methods. The latter can be subdivided into isotonic, hypertonic and hypotonic systems. In most cases the aqueous isolation renders nuclei closer to their physiological status in the cell. A standard method for the hypotonic isolation of nuclei is presented and the methodology of nuclear isolation is discussed.

Aedes↗

Karyophilic properties of Semliki Forest virus nucleocapsid protein.

Semliki Forest virus capsid (C) protein molecules (Mr, 33,000) can be introduced efficiently into the cytoplasm of various target cells by electroporation, liposome, and erythrocyte ghost-mediated delivery (M. Elgizoli, Y. Dai, C. Kempf, H. Koblet, and M.R. Michel, J. Virol. 63:2921-2928, 1989). Here, we show that the transferred C protein molecules partition rapidly from the cytosolic compartment into the nucleus. Transport of the C protein molecules into the nucleus was reversibly arrested by metabolic inhibitors, indicating that the transfer process is energy dependent. Fractionation of isolated nuclei revealed that the delivered C protein preferentially associates with the nucleoli. This finding was confirmed by morphological studies, showing that in an in vitro system containing ATP isolated nuclei rapidly accumulated rhodamine-labeled C protein in their nucleoli. Furthermore, in this assay system, the lectin wheat germ agglutinin prevented transfer of C protein through nuclear pores. These results are in agreement with our observation that nucleoli contain measurable amounts of newly synthesized C protein as early as 5 h after infection of cells with SFV. Thereafter, nucleolar-associated C protein increased progressively during the course of infection.

Adenosine Triphosphate↗

[An automated in vitro toxicity test of 25 chemicals].

A computer controlled bench-top analyser was used to establish an automated in vitro-cytotoxicity test. It is based on the continuous monitoring of growth of Salmonella typhimurium TA 98 and TA 100 (strains that are also employed in the classical Ames test (1) for mutagenicity screening) under the influence of the toxic compounds. With this method we can quantify general cytotoxicity and in addition, some questions concerning the mechanism of the toxic influence on the test organisms can be answered. Its simplicity and reliability as well as the good correlation with in vivo data make the new test system suitable for pre-screening the toxicity of a large number of compounds within short time.

Biological Assay↗

Automated turbidimetry for the rapid differentiation and enumeration of bacteria in food.

A fully automated analysis system has been used to determine bacterial contamination in food, both qualitatively and quantitatively. Growth curves of bacterial flora were monitored using differential media, and a correlation was established between growth curve parameters and colony-forming units derived from drop-plating as the reference method. Optimal correlation was reached by multiple linear regression. The results were already obtained 16 h.

Animals↗