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Biomedical subjects

T Jelinek

Publications and source records attributed to T Jelinek.

At least 91 records · Page 5Linked to original sources
PubMed1995

Biochemical analysis of MEK activation in NIH3T3 fibroblasts. Identification of B-Raf and other activators.

Numerous potential activators of MEK have been identified, including c-Raf-1, B-Raf, c-Mos, and a family of MEK kinases. However, little information gives insight into the activators actually utilized in vivo. To address this, we have used column chromatography and a coupled MEK activation assay to identify in NIH3T3 cells, two major MEK activators, and a third insulin-specific activator. The first MEK activator has an apparent M(r) of 40,000-50,000, was immunologically distinct from A-Raf, B-Raf, c-Raf-1, c-MEKK, c-Mos, MEK1, and MEK2, and was rapidly activated by serum, platelet-derived growth factor (PDGF), insulin, thrombin, and phorbol ester. The second MEK activator was identified as B-Raf. Activation of 93-95 kDa B-Raf was observed in column fractions and B-Raf immunoprecipitates from cytosolic and particulate fractions after stimulation with serum or PDGF, but not insulin. c-Raf-1 from cytosol did not exhibit MEK activator activity; however, c-Raf-1 immunoprecipitates from the particulate fraction revealed MEK activator activity that was enhanced after stimulation with PDGF or phorbol ester, but not serum or insulin. Both c-Mos and c-MEKK were present in NIH3T3 fibroblasts but did not show MEK activator activity. These data provide direct evidence that 93-95-kDa B-Raf isozymes and an unidentified 40-50-kDa MEK activator are major agonist-specific MEK activators in NIH3T3 fibroblasts.

3T3 Cells↗
PubMed1995

Cutaneous myiasis: review of 13 cases in travelers returning from tropical countries.

BACKGROUND: Cutaneous myiasis is frequently reported in patients from tropical countries. Most commonly infestation is due to Cordylobia anthropophaga and Dermatobia hominis, whereas Hypoderma lineatum is less likely to affect humans. METHODS: The clinical and parasitologic aspects in a series of 13 patients with cutaneous myiasis seen in a German travel clinic are reviewed. RESULTS: Six patients were infected with larvae of the tumbu fly, Cordylobia anthropophaga, six with larvae of the botfly, Dermatobia hominis, and one patient presented with an infection with Hypoderma lineatum. Dermatobia hominis was exclusively acquired in the American tropics and Cordylobia anthropophaga exclusively in Africa, whereas infection with Hypoderma lineatum was acquired in Nepal. The number of larvae removed from the skin was markedly higher in patients infected with the tumbu fly (average of five larvae) compared to the botfly (average of 1.7 larvae), whereas pain was more frequently reported from patients with botfly infestation. Treatment consisted in the removal of the larvae by forceps or surgical incision in tumbu or botfly infection and oral application of ivermectin in infection with Hypoderma lineatum. This led to complete recovery in all patients. CONCLUSIONS: In patients presenting with cutaneous lesions after return from tropical countries, myiasis has to be considered as a differential diagnosis. Simple and effective therapy is available through removal of the maggots.

Adult↗
PubMed1995

Long-term efficacy of primaquine in the treatment of vivax malaria in nonimmune travelers.

Fifty-six travelers presenting with vivax malaria to a German travel clinic were followed regularly for at least 18 months between 1984 and 1992 to investigate the long-term efficacy of primaquine in nonimmune patients without reinfection. All received a standard treatment of 15 mg of primaquine a day for 14 days following an initial total dose of 1,500 mg of chloroquine (base) given over a 48-hr period. None of the patients visited countries endemic for malaria during the period of observation. In seven patients (12.5%), relapses were confirmed microscopically by detection of malaria parasites in blood films. The frequency of relapses varied between one and four per patient and these occurred between 60 and 252 days after treatment. Four of these seven patients had acquired infection in Papua New Guinea or eastern Indonesia, while only five (10.2%) of the remaining 49 patients without relapses had traveled to these areas prior to referral (P < 0.01).

Adolescent↗
PubMed1995

Quinine resistant falciparum malaria acquired in east Africa.

A 43 year old man with falciparum malaria acquired in East Africa was treated with quinine intravenously at a loading dose of 500 mg and subsequently 500 mg tid. Within 42 hours after initiation of treatment the parasitaemia increased from 2% to 16%. A RIII-resistance against quinine was suspected and therapy was switched to oral administration of halofantrine (500 mg at 6 hourly intervals) which led to complete recovery. Blood samples were cultured for malaria parasites 42 hours after start of therapy with quinine but before initiation of therapy with halofantrine. In vitro resistance testing was performed with samples directly derived from the patient and after 24 and 48 hours of culturing. In repeated tests an in vitro resistance to quinine could be confirmed (IC50: 25.6 x 10(-6) mol/l, IC99: > 51.2 x 10(-6) mol/l) while the strain was fully susceptible to chloroquine (IC50: < 0.4 x 10(-6) mol/l, IC99: 1.6 x 10(-6) mol/l), mefloquine (IC50: < 0.4 x 10(-6) mol/l, IC99: 3.2 x 10(-6) mol/l), tetracycline (IC50: 0.16 x 10(-6) mol/l, IC99: 0.32 x 10(-6) mol/l) and halofantrine (IC50: 0.02 x 10(-6) mol/l, IC99: 0.04 x 10(-6) mol/l). Increased susceptibility to quinine after addition of verapamil was noted. The presence of a specific mutation, on the pfmdr1-gene on chromosome 5, previously associated with chloroquine drug resistance, could be confirmed by polymerase chain reaction. To our knowledge a R III-in vivo and in vitro resistance of Plasmodium falciparum to quinine has not been described yet in East Africa.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Evaluation of circumsporozoite antibody testing as a sero-epidemiological tool for the detection of Plasmodium falciparum infection in non-immune travelers.

The objective of this investigation was to collect data concerning CS-antibody levels and duration of the immunological response to exposure of non-immune persons to a single malaria infection. For this purpose 156 specimens from 98 patients with confirmed falciparum malaria, 76 specimen derived from 64 patients with vivax malaria and sera from 32 patients who had not been previously to malarious areas were investigated by use of a commercially available ELISA testkit. All specimens from patients with falciparum malaria were also tested for merozoite-antibodies by an indirect fluorescence antibody test (IFAT). Positive levels of merozoite-antibodies were detectable in 89.1% of the specimen in this panel during the period between days 8 and 90 after onset of symptoms and decreased steadily thereafter. The test results were positive for CS-antibodies in 36.4% of the specimens from patients with falciparum malaria during the first 7 days after onset of symptoms. This figure increased to 55.8% during days 8-90 after onset and decreased to 38.9% in specimens which were tested later (91-1898 days). 11 specimens reacted positively to CS-antibody testing but negative in the IFAT. Therefore, the percentage of specimen detected by either IFAT or CS-ELISA was at 51.9% during days 0 and 7 (p < 0.001), 95.3% during days 8 and 90 (p = 0.039) and 44.4% for testing performed later (p < 0.001). CS-antibodies could also be detected in 5.3% of specimen from patients with vivax malaria while none of the sera from the malaria-negative control-group tested positive for CS-antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Protozoan↗
PubMed1995

Stand-by treatment of suspected malaria in travellers.

Travellers to malarious areas are increasingly advised to carry an emergency medication for self-treatment of suspected malaria in absence of medical attention. However, no data are available so far how travellers cope with self-diagnosis and stand by treatment (SBT). We therefore investigated the frequency, circumstances and outcome of emergency self-treatment for suspected malaria in German travellers. 3434 travellers were recruited for an open prospective study by 28 different travel clinics in Germany. 2867 travellers (90.1%) who returned questionnaires after their journey were analyzed. 40 travellers (1.4%) reported about SBT during their journey. Significant Plasmodium falciparum antibody levels could be demonstrated in only 4 of 37 SBT users (10.4%). In another 127 travellers with febrile episodes but without SBT use, no malaria was indicated by follow-up and/or serology.

Adult↗
PubMed1994

Malaria in Nonimmune Travelers: A Synopsis of History, Symptoms, and Treatment in 160 Patients.

With the current increase of international travel to tropical endemic areas, the incidence of malaria being imported into nonendemic countries has increased significantly. Disagreement concerning malaria chemoprophylaxis and inadequate knowledge of malarious areas, morbidity, and pretravel advise has led to confusion among both health professionals as well as travelers. Therefore, this study was conducted to investigate malaria imported into Germany by identifying the high-risk endemic areas, clinical presentations, and chemoprophylactic and therapeutic regimens related to reported cases. Between 1990 and 1993, the 160 nonimmune travelers, all German nationals or residents for more than 10 years, presenting to our travel clinic with microscopically confirmed malaria were investigated. For each, the travel history, chemoprophylaxis used during travel, symptoms, pathological diagnosis, and treatment efficacy were analyzed. Africa (73%), Asia (21%), and Central South America (6%) were the endemic countries visited by our patients, of whom only 3% used the chemoprophylaxis recommended for their destination. Plasmodium falciparum was the most common pathogen, found in more than half of our patients, and P. vivax (29%), P. ovale (6%), P. malariae (6%), a mixed infection with P. falciparum and P. vivax (3%) were also detected. All patients presented with fever and headaches, a majority with profuse night sweats, insomnia, arthralgias, and myalgias, and diarrhea and abdominal cramps were experienced in 13% and 8%, respectively. In falciparum malaria, a recrudescence was observed in all patients who received chloroquine only, whereas quinine, halofantrine, and mefloquine were highly effective. In vivax malaria, a relapse rate of 14% was noted in patients treated with the currently recommended regimen of chloroquine and primaquine. Visitors to endemic countries, especially Africa, are of significant risk. Given the low compliance rate of chemoprophylaxis, a high precentage of malaria in our patients could have been avoided by an appropriate prophylaxis regimen and optimal pretravel counseling.

Journal Article↗
PubMed1994

[Gnathostomiasis after a stay in Thailand].

During a 4-week holiday in Thailand a 13-year-old boy from Germany briefly fell ill with diarrhoea without blood-tinged stools and fever. 5 months later oedematous, reddish swellings occurred, at first on the right upper arm, later over the right side of the chest. Especially at night these swellings were associated with severe local and generalized itching. Bilaterally the axillary lymph-nodes were hard and slightly enlarged. There was a marked eosinophilia of 56%, the white count was 22,700/microliters and the serum level of IgE was raised to 1777 IU/ml. No worm eggs or larvae were found in the stool. Enzyme-linked immunosorbent assay, using larvae of Gnathostoma spinigerum as antigen, demonstrated antibodies against this nematode. Gnathostomiasis having been diagnosed treatment was started with albendazole (200 mg twice daily for 21 days), after which the boy was symptom free. After 2 months the eosinophilia had regressed to 10%, the white cell count was normal and the antibody titre had fallen.

Adolescent↗
PubMed1994

c phosphorylation and activation of the IGF-I receptor in src-transformed cells.

Using a panel of src mutants partially defective for malignant transformation, our laboratory has previously identified the insulin-like growth factor (IGF-I) receptor as a protein whose tyrosine phosphorylation correlates with transformation by src in embryonic chick cells (Kozma et al., 1990; Kozma and Weber, 1990). It has not been clear, however, whether src-induced phosphorylation altered the enzymatic or signaling properties of the IGF-I receptor and thus whether the IGF-I receptor could be a functionally significant target for pp60v-src. To examine the effect of src expression on the activity of the IGF-I receptor, the human IGF-I receptor was expressed in Rat-1 fibroblasts co-expressing the temperature-sensitive v-src mutant, tsLA29. The IGF-I receptor exhibited an elevated level of tyrosine phosphorylation in src transformed cells even in the absence of IGF-I treatment. Increased receptor phosphorylation occurred rapidly when cells expressing a temperature-conditional src mutant were shifted from the restrictive to the permissive temperature. Src-induced phosphorylation of the receptor was correlated with an increase in the in vitro tyrosine kinase activity of the receptor, both toward itself and exogenous substrates. The src-induced increase in receptor activity was shown to be dependent on tyrosine phosphorylation, as treatment with a tyrosine-specific phosphatase lowered receptor activity. A kinase-defective mutant of the IGF-I receptor also became constitutively phosphorylated in src-transformed cells, ruling out a possible autocrine mechanism for this phosphorylation. Collectively these data indicate that pp60v-src induces ligand-independent phosphorylation and activation of the IGF-I receptor by an intracellular mechanism, consistent with the possibility that receptor phosphorylation could contribute to the genesis of the transformed phenotype.

Animals↗
PubMed1994

Cutaneous larva migrans in travelers: synopsis of histories, symptoms, and treatment of 98 patients.

The symptoms, medical history, and treatment of 98 patients with cutaneous larva migrans (creeping eruption) who attended a travel-related-disease clinic during a period of 4 years are reviewed. This condition is caused by skin-penetrating larvae of nematodes, mainly of the hookworm Ancylostoma braziliense and other nematodes of the family Ancylostomidae. Despite the ubiquitous distribution of these nematodes, in the investigated group only travelers to tropical and subtropical countries were affected; 28.9% of the patients had symptoms for > 1 month, and for 24.5% the probable incubation period was > 2 weeks. The efflorescences typically were on the lower extremities (73.4% of all locations). The buttocks and anogenital region were affected in 12.6% of all locations, and the trunk and upper extremities each were affected in 7.1%. Only a minority of patients presented with eosinophilia or an elevated serum level of IgE. No other laboratory data appeared to be related to the disease. Therapy with topical thiabendazole was successful for 98% of the patients. Systemic antihelmintic therapy was necessary in two cases because of disseminated, extensive infection.

Adult↗
PubMed1994

Tumorigenicity of adenovirus-transformed rodent cells is influenced by at least two regions of adenovirus type 12 early region 1A.

Chimeric adenovirus type 5 (Ad5)/Ad12 early region 1A (E1A) genes were used to transform primary baby rat kidney cells in cooperation with Ad12 E1B, and the resulting cell lines were assayed for tumorigenicity in syngeneic rats. It was found that lines were nontumorigenic when transformed by hybrid E1A genes consisting of the amino-terminal 80 amino acids from Ad12 including conserved region 1 (CR1), with the remaining portion from Ad5. In contrast, cell lines transformed by hybrids containing Ad12 E1A sequences from the amino terminus to the leftmost border of CR3 or beyond were tumorigenic. To extend these results, sequences spanning CR2 and CR3 of Ad5 E1A were replaced with the homologous regions of Ad12 E1A and additional transformed cell lines were established. These lines were weakly-to-moderately tumorigenic, suggesting that Ad12 E1A sequences between CR2 and CR3 may be involved in tumorigenicity but are not the sole factors influencing it. Interestingly, examination of an E1A sequence alignment indicated that the region between CR2 and CR3 of Ad12 E1A is also conserved in the corresponding sequence of simian adenovirus type 7, which, like Ad12, is highly oncogenic. This region is characterized by the presence of a stretch of several alanine residues and is similar to a motif present in a number of proteins with transcriptional repression activity. The possibility that this region may influence tumorigenicity by means of a transcriptional regulatory mechanism is discussed.

Adenovirus E1A Proteins↗
PubMed1994

RAS and RAF-1 form a signalling complex with MEK-1 but not MEK-2.

Recent studies have demonstrated the existence of a physical complex containing p21ras (RAS), p74raf-1 (RAF-1), and MEK-1. Although it is clear that formation of this complex depends on the activation state of RAS, it is not known whether this complex is regulated by the activation state of the cell and whether MEK-2 is also present in the complex. To analyze the regulation and specificity of this complex, we utilized immobilized RAS to probe lysates of cultured NIH 3T3 fibroblasts and analyzed the proteins complexing with RAS following serum starvation or stimulation. Complex formation among RAS, RAF-1, and MEK-1 was dependent only on RAS:GMP-PNP and not on cell stimulation. Incubations of lysates with immobilized RAS depleted all RAF-1 from the lysate but bound only a small fraction of cytosolic MEK-1, and further MEK-1 could bind immobilized RAS only if exogenous RAF-1 was added to the lysate. This indicates that binding of MEK-1 to RAS depends on the presence of RAF-1 or an equivalent protein. In contrast to MEK-1, MEK-2 was not detected in the RAS signalling complex. A proline-rich region of MEK-1 containing a phosphorylation site appears to be essential for signalling complex formation. Consistent with the preferential binding of MEK-1 to RAS:RAF-1, the basal activity of MEK-1 in v-ras-transformed cells was found to be elevated sixfold, whereas MEK-2 was elevated only twofold, suggesting that the RAS signalling pathway favors MEK-1 activation.

3T3 Cells↗
PubMed1994

Human infection with Gongylonema pulchrum: a case report.

A 43 year old woman developed a painful tumor at the left buccal mucosa. Following local anti-inflammatory treatment a 35 mm long, living female adult worm of Gongylonema pulchrum was extracted from the affected side. No further treatment was needed and recovery was complete 5 days after extraction. Infection had occurred possibly 6 weeks before in Hungary with ingestion of contaminated water from an open draw well. Although commonly occurring as parasitic infection of domestic cattle and other vertebrates, gongylonemiasis is very rare in humans. Only 48 cases have been described in the literature since 1864. Life cycle and pathology of G. pulchrum are discussed.

Adult↗
PubMed1993

Inhibition of the EGF-activated MAP kinase signaling pathway by adenosine 3',5'-monophosphate.

Mitogen-activated protein (MAP) kinases p42mapk and p44mapk are activated in cells stimulated with epidermal growth factor (EGF) and other agents. A principal pathway for MAP kinase (MAPK) activation by EGF consists of sequential activations of the guanine nucleotide exchange factor Sos, the guanosine triphosphate binding protein Ras, and the protein kinases Raf-1, MAPK kinase (MKK), and MAPK. Because adenosine 3',5'-monophosphate (cAMP) does not activate MAPK and has some opposing physiologic effects, the effect of increasing intracellular concentrations of cAMP with forskolin and 3-isobutyl-1-methylxanthine on the EGF-stimulated MAPK pathway was studied. Increased concentrations of cAMP blocked activation of Raf-1, MKK, and MAPK in Rat1hER fibroblasts, accompanied by a threefold increase in Raf-1 phosphorylation on serine 43 in the regulatory domain. Phosphorylation of Raf-1 in vitro and in vivo reduces the apparent affinity with which it binds to Ras and may contribute to the blockade by cAMP.

1-Methyl-3-isobutylxanthine↗
PubMed1992

Recombinant human adenoviruses containing hybrid adenovirus type 5 (Ad5)/Ad12 E1A genes: characterization of hybrid E1A proteins and analysis of transforming activity and host range.

Hybrid adenovirus type 12 (Ad12)/Ad5 E1A genes were constructed by homologous recombination in Escherichia coli, a technique which offers several advantages over conventional mutagenesis for genetic analysis of proteins. In particular, functional differences between the proteins can be mapped by correlating the replacement of specific sequences with the acquisition of new properties, and there is no requirement for common unique restriction sites or polymerase chain reaction strategies to construct the hybrids. Recombinant adenoviruses expressing these hybrid E1A proteins were capable of replicating efficiently in HeLa cells, with the exception of one construct which contained a hybrid transactivation domain. The transforming activity of the hybrid E1A constructs was assayed by DNA transfection of primary baby rat kidney cells. Plasmids containing Ad12 E1 were approximately 20-fold less efficient at transformation than those with E1 of Ad5, and it was found that two regions in exon 1 of E1A mediate this difference. No differences were found in the abilities of any hybrid E1A proteins to bind to cellular proteins previously determined to be important for transformation by E1A.

Adenovirus Early Proteins↗
PubMed1990

Chromosomal damage induced by human adenovirus type 12 requires expression of the E1B 55-kilodalton viral protein.

Infection of human embryonic kidney cells with adenovirus type 12 results in the induction of damage at specific (17q21-22, 1p36, 1q21, and 1q42-43) and random sites in the cellular chromosomes. A previous study by Durnam et al. (D. M. Durnam, P. P. Smith, J. C. Menninger, and J. K. McDougall, Cancer Cells 4:349-354, 1986) indicated that the expression of viral early region 1 (E1) is sufficient for the induction of damage at band 17q21-22. In the present report we used an adenovirus type 12-adenovirus type 5 recombinant with E1A hybrid sequences as well as viruses with mutations in the adenovirus type 12 E1B genes to map adenovirus type 12 E1 functions involved in the induction of genetic damage. Our results show that the expression of the E1A proteins is not sufficient for this effect. On the other hand, mutations within the E1B 55-kilodalton protein but not the E1B 19-kilodalton protein affect the ability of the virus to induce both specific and random chromosomal damage.

Adenoviruses, Human↗