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Biomedical subjects

A Dietrich

Publications and source records attributed to A Dietrich.

At least 55 records · Page 3Linked to original sources

Evolution of a mitochondrial tRNA PHE gene in A. thaliana: import of cytosolic tRNA PHE into mitochondria.

Previously we have described a putative tRNATyr in Arabidopsis thaliana mitochondria, the sequence of which is different from that of other plant mitochondrial tRNATyr genes. We show here that this tRNATyr gene sequence is present in several copies in the mitochondrial genome of A. thaliana. One copy of these tRNATyr gene sequences, termed here tRNATyr-1, could encode a functional tRNA. Expression analysis has shown that the tRNATyr-1 gene is cotranscribed with the downstream tRNAGlu gene, and that the corresponding mature-sized tRNA is present in mitochondria. We also show that the native tRNATyr gene, similar to the mitochondrial tRNATyr genes found in plants, is present in the A. thaliana mitochondrial genome and expressed. The tRNATyr-1 gene has been previously suggested to be derived from a tRNAPhe gene sequence. We show here that, as a consequence, there is no tRNAPhe gene in the mitochondrial genome of A. thaliana and that a cytosolic tRNAPhe is imported in A. thaliana mitochondria.

Arabidopsis↗

Evolutionary aspects of "chloroplast-like" trnN and trnH expression in higher-plant mitochondria.

Two identical "chloroplast-like" tRNAAsn genes, trnN1 and trnN2, have been identified in the potato (Solanum tuberosum) mitochondrial genome. The flanking sequences of trnN1 are unrelated to the corresponding authentic potato chloroplast regions, whilst those of trnN2 are very similar to the chloroplast sequences. The trnN1 copy is present in the mitochondrial genome of various plants whereas the second copy, trnN2, is absent from all the other plant genomes studied so far. Interestingly, both trnN copies are expressed in potato mitochondria. Sequences flanking the chloroplast-like tRNAHis gene (trnH), present as a single copy in the potato mitochondrial DNA, are unrelated to the corresponding chloroplast sequences, whereas chloroplast-derived sequences have been maintained in the vicinity of the maize chloroplast-like mitochondrial trnH gene. However, both the potato and the maize trnH are expressed in mitochondria.

Base Sequence↗

Vasopressin and memory. I. The vasopressin analogue AVP4-9 enhances working memory as well as reference memory in the radial arm maze.

The present study examined the effects of vasopressin on memory. Healthy rats were injected with the arginine vasopressin fragment AVP4-9 and the AVP antagonist [beta-mercapto-beta,beta-cyclopentamethylenepropionyl1,O-Et-Pyr 2,Val4,Arg8] and were tested in an eight-arm radial maze for 60 sessions. All injections were given s.c. 30 min prior to testing. AVP4-9 enhanced radial arm maze performance. AVP4-9 treated animals showed enhancement in performance as well as increases in the rate of learning, indicating that they learned the task faster. Furthermore, the overall memory enhancement was due to improved working memory as well as to improved reference memory. These results cannot be explained in terms of changes in locomotor activity because an open field test revealed no differences between groups for both of these compounds. The AVP antagonist did not impair performance in the radial maze. It is concluded that AVP4-9 has a more general effect on memory, one that is not limited to a specific type of memory.

Animals↗

Vasopressin and memory. II. Lesions to the hippocampus block the memory enhancing effects of AVP4-9 in the radial maze.

Previous work has shown that the arginine vasopressin fragment AVP4-9 enhanced overall performance in the radial arm maze. This enhancement effect was due to improved working memory as well as to improved reference memory. The present study uses a behavioral approach to investigate whether these effects of vasopressin on memory are mediated by the hippocampus. Animals received either NMDA lesions to the hippocampus or sham operations and were then treated with AVP4-9 or saline. The testing was performed in an eight-arm radial maze for 60 sessions, and all injections were given s.c. 30 min prior to testing. The hippocampal lesioned animals showed a marked deficit in working memory that was not ameliorated by AVP4-9; however, the improvement in reference memory produced by the compound was as large as in healthy animals. Since the AVP treatment did not differentiate between the two hippocampal groups, the memory enhancing effects of AVP4-9 were blocked by hippocampal lesions. It is concluded that vasopressin has a more general effect on memory and that its site of action includes but is not limited to the hippocampus.

Animals↗

Protection from acetaminophen-induced liver damage by the synergistic action of low doses of the poly(ADP-ribose) polymerase-inhibitor nicotinamide and the antioxidant N-acetylcysteine or the amino acid L-methionine.

1. An array of therapeutically used analgetic and antirheumatic drugs cause severe liver damage. The present study investigates the hepatoprotective effects of inhibitors of NAD-dependent adenoribosylation reactions and of antioxidants in analgesic-induced hepatic injury. 2. Male NMRI mice were treated PO with 500 mg/kg of acetaminophen, and the activities of both glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) were determined in serum. 3. The acetaminophen-induced release of both GOT and GPT from injured liver cells could be inhibited in a dose-dependent manner, when mice were injected additionally either with increasing amounts (from (25 mg/kg to 100 mg/kg i.p.) of the PARP-inhibitor nicotinamide, with increasing amounts (from 25 mg/kg to 100 mg/kg i.p.) of the antioxidant N-acetylcysteine, or with increasing amounts (from 50 mg/kg to 300 mg/kg i.p.) of the amino acid L-methionine. 4. A combination of both nicotinamide and N-acetylcysteine (at the low dose of 12.5 mg/kg i.p. each) results in a complete protection from acetaminophen-induced release of GOT and GPT from injured liver cells. 5. A combination of both L-methionine and N-acetylcysteine or nicotinamide (at the low dose of 12.5 mg/kg IP each) resulted also in complete protection from acetaminophen-induced release of GOT and GPT.

Acetaminophen↗

Suppression of type II collagen-induced arthritis by N-acetyl-L-cysteine in mice.

1. The antiarthritic and anti-inflammatory efficacy of N-acetyl-L-cysteine (NAC) was tested in male DBA/1 hybrid mice suffering from type II collagen-induced arthritis. Parameters including the arthritis index and the phagocytic responses recorded by chemiluminescence in unseparated blood were used for the assessment of disease activity. 2. Mice were immunized by subdermal injection of bovine type II collagen in Freund's complete adjuvant. The treatment with NAC started at day 42 after immunization and was continued over a period of six weeks: in doses ranging up to 50 mg/kg, a dose-dependent suppression of arthritis was noted; between 50 and 200 mg/kg, the inhibition curve had a plateau [ED50 = 50 mg/(kg x day)]. 3. The arthritis index correlated positively with the generation of chemiluminescence by reactive oxygen species (ROS) produced in neutrophils and monocytes activated by 12-O-tetradecanoylphorbol 13-acetate. 4. After treatment with 100 mg/kg of NAC from day 42 after immunization over a period of six weeks, the ROS production was reduced to levels occurring in whole blood of healthy animals. 5. It is concluded that low-molecular-weight antioxidants such as NAC may be adequate for controlling oxidative stress-derived damage in rheumatic diseases by modulation of ROS-dependent signal transduction pathways.

Acetylcysteine↗

High CD44 surface expression on primary tumours of malignant melanoma correlates with increased metastatic risk and reduced survival.

The cell surface glycoprotein CD44 has been implicated in the progression and metastasis of certain human tumours including malignant melanoma (MM). In animal models, certain MM cell lines, expressing high levels of CD44, displayed an augmented capacity for haematogenous metastasis, compared to those with low CD44 levels. To determine whether, in vivo, the level of CD44 expressed by primary tumours of MM (PMM) is related to their metastatic potential, CD44 expression on PMM was studied in 92 patients, classified by their metastatic risk based on histological measurement of vertical tumour thickness (VT): in situ PMM, low-risk PMM (VT < or = 0.7 mm), intermediate risk PMM (VT = 0.71-1.4 mm) and high-risk PMM (VT > 1.4 mm). Paraffin-embedded sections were stained immunohistochemically with a panCD44 MAb. The level of CD44 expression on PMM was analysed semiquantitatively with epidermal CD44 staining set as an internal standard. High levels of CD44 were detected in 58.3% of high-risk PMM, 40.6% of intermediate-risk PMM, 36.7% of low-risk PMM and 16.7% of in situ PMM. Seventy-four per cent (17/23) of patients who developed and/or died of MM metastasis were CD44 high, and importantly, among these were 5 patients, whose metastatic risk had been estimated low, based on the measurement of VT. Finally, Kaplan-Meier analysis revealed patients whose PMM were CD44 high to have a significantly reduced 5-year survival rate compared to those that were CD44 low (P < 0.05). We conclude that in our patient population, a high level expression of CD44 on PMM is associated with increased metastatic risk and reduced survival.

Antigens, Neoplasm↗

Detection of CD44 splice variants in formalin-fixed, paraffin-embedded specimens of human skin cancer.

The standard form of CD44 (CD44s) and CD44 isoforms, containing sequences encoded by one or several of 10 different variant CD44 exons (v1-v10), are thought to play a crucial role in the growth and metastasis of certain human tumors. Recently, monoclonal antibodies (mAbs) directed against all CD44 isoforms (panCD44), or against epitopes encoded by specific variant exons (CD44v) have been developed, which unfortunately only stain cryopreserved tissues. We wished to develop a technique to unmask chemically CD44s and CD44v epitopes in paraffin-embedded specimens of human skin cancers, so that they would be accessible for these mAbs. To address this issue, CD44s and CD44v expression was compared in cryopreserved and in formalin-fixed, paraffin-embedded biopsies obtained from the same basal cell carcinomas (BCC), squamous cell carcinomas (SCC), primary malignant melanomas (PMM) and metastatic malignant melanomas (MMM). Formalin-fixed tumors were deparaffinized and treated briefly with an antigen retrieval fluid (TUFTM) at 95 degrees C or left untreated. In untreated paraffin-embedded tissues, no CD44s or CD44v staining was detected. In contrast, in antigen retrieval fluid-treated biopsies CD44s and CD44v expression was identical to that in cryopreserved specimens of the same tumor with the exception of mAbs detecting v7/8 and v10. We conclude that antigen retrieval unmasks certain epitopes in formalin-fixed, paraffin-embedded tissues, thus facilitating future research on the relevance of CD44s and CD44v expression for human skin carcinogenesis.

Alternative Splicing↗

Is the hippocampus involved in temporal discrimination and the memory of short intervals?

Rats with lesions to the hippocampus proper and the subiculum were tested for timing behavior and temporal memory. Using the peak procedure, they were trained to discriminate a 40 s interval and a retention gap tested the memory for time. Results were interpreted within the theoretical framework of the internal clock and with respect to current theories on hippocampal function. Timing behavior was unaffected by either lesion and no shifts in the temporal discrimination functions were observed. The lesions also failed to show a deficit in the memory for temporal events. For all groups, the retention gap increased the mean peak time by the time of the gap. This indicated that all rats used the stop rule which required the use of working memory. Thus, it was concluded that the hippocampus is neither necessary for accurate timing behavior nor for the memory of temporal events.

Animals↗

Impact of growth factors on morphometric corneal endothelial cell parameters and cell density in culture-preserved human corneas.

PURPOSE: Donor corneas can be preserved for < or = 4 weeks in organ culture (31 degrees C) by using modified minimal essential medium (MEM). About one fifth of them have to be discarded, however, as disintegration of the endothelial cell monolayer-enhanced polymegethism, cell loss-occurs. The objective of this study was to investigate whether addition of insulin, dextran, and epidermal growth factor (EGF) makes corneal endothelial cells more viable, stable, and homogeneous. METHODS: Sixteen paired human donor corneas were cocultured in media supplemented with EGF and insulin-like growth factor (IGF) or in conventional modified MEM for 4 weeks. Endothelial parameters were evaluated at the outset and at days 7, 14, 21, and 28 of culture by using an automated digital image-analysis system. RESULTS: No significant differences were observed in the first 2 weeks of culture. Beginning with day 14, however, stabilization of endothelial cell patterns was evident for corneas cultured in supplemented culture media. CONCLUSION: Our data indicate that the addition of growth factors to culture media might increase the percentage of corneas available for transplantation and would also allow a significantly longer period of preservation.

Cell Count↗

German rescue system--an overview.

Aim of the report is to show the structure, organisation and legal aspects of the rescue system of the Federal Republic of Germany. The laws are the basis of the rescue system in each of the sixteen federal countries. The governments of the countries delegate their tasks to the administrative districts. Several organisations like German Red Cross, the Fire Brigades; societies like Arbaite-Samariter-Bund, Johanniter-Unfall-Hilfe, Malteser-Hilfsdienst take part in the rescue system with a great number of paid professional people and volunteers with a more or less professional education. Who is Who in the rescue system? Who is responsible for medical education and what is the difference between the rescue system and the "doctor on emergency call"? Both systems exist side by side, but they are very different from one another. The "doctor on emergency call" is private organised by the board of general practitioners and is responsible for "all day" emergency cases on weekends and during the night. The task of the countries and districts is to care for the rescue services, and they should only be used for severe injuries and illness. Most of the physicians taking part in the rescue services belong to hospitals and have no financial interests in their duty.

Emergency Medical Services↗

Isoprenylation of the G protein gamma subunit is both necessary and sufficient for beta gamma dimer-mediated stimulation of phospholipase C.

We have previously shown that isoprenylation and/or additional post-translational processing of the G protein gamma 1 subunit carboxyl terminus is required for beta 1 gamma 1 subunit stimulation of phospholipase C-beta 2 (PLC beta 2) [Dietrich, A., Meister, M., Brazil, D., Camps, M., & Gierschik, P. (1994) Eur. J. Biochem. 219, 171-178]. To examine whether isoprenylation of the gamma 1 subunit alone is sufficient for beta 1 gamma 1-mediated PLC beta 2 stimulation or whether any of the two subsequent modifications, proteolytic removal of the carboxyl-terminal tripeptide and/or carboxylmethylation, is required for this effect, nonisoprenylated recombinant beta 1 gamma 1 dimers were produced in baculovirus-infected insect cells, purified to near homogeneity, and then isoprenylated in vitro using purified recombinant protein farnesyltransferase. Analysis of the beta 1 gamma 1 dimer after in vitro farnesylation by reversed phase high-performance liquid chromatography followed by delayed extraction matrix-assisted laser desorption/ionization mass spectrometry confirmed that the gamma 1 subunit was carboxyl-terminally farnesylated but not proteolyzed and carboxylmethylated. Functional reconstitution of in vitro-farnesylated beta 1 gamma 1 dimers with a recombinant PLC beta 2 isozyme revealed that farnesylation rendered recombinant nonisoprenylated beta 1 gamma 1 dimers capable of stimulating PLC beta 2 and that the degree of this stimulation was only approximately 45% lower for in vitro-farnesylated beta 1 gamma 1 dimers than for fully modified native beta 1 gamma 1 purified from bovine retinal rod outer segments. Taken together, these results suggest that isoprenylation of the gamma subunit is both necessary and sufficient for beta gamma dimer-mediated stimulation of phospholipase C.

Alkyl and Aryl Transferases↗

Electroporation of small RNAs into plant protoplasts: mitochondrial uptake of transfer RNAs.

To study tRNA import into plant mitochondria, we have set up a system to follow the fate in vivo of tRNA transcripts introduced into plant protoplasts by electroporation. Conditions were optimized for maximum tRNA uptake into potato protoplasts. We have shown that in vitro synthesized tRNA transcripts are poor substrates due to rapid degradation leading to low efficiencies of transfer and short life in protoplasts. Labelled natural tRNAs were more efficiently electroporated into protoplasts and they remained stable during protoplast culture. We have observed import into mitochondria of total and purified cytosolic tRNAs in protoplasts but the process was not specific for the tRNA species which are normally imported.

Cells, Cultured↗

Synergistic effects of thalidomide and poly (ADP-ribose) polymerase inhibition on type II collagen-induced arthritis in mice.

The present study investigates synergistic effects of the TNF-alpha inhibitor thalidomide and the poly(ADP-ribose) polymerase (PARP)-inhibitor nicotinic acid amide (NAA) in male DBA/1 hybird mice suffering from type II collagen-induced arthritis. Parameters including the arthritis index, chemiluminescence and anti-collagen antibody titers were used for the assessment of disease activity: The disease courses demonstrated clearly an inhibitory effect of thalidomide. NAA inhibited established collagen arthritis in a dose-dependent manner. The combined application of thalidomide and NAA caused a powerful synergistic inhibition of arthritis. Furthermore, thalidomide and NAA were tested ex vivo for their inhibition of the NADPH oxidase-dependent generation of reactive oxygen species by activated neutrophils and monocytes in unseparated human blood. Our data show that type II collagen-induced arthritis can be suppressed by the simultaneous inhibition of TNF-alpha, PARP, and NADPH oxidase.

Animals↗

Editing and import: strategies for providing plant mitochondria with a complete set of functional transfer RNAs.

The recombinations and mutations that plant mitochondrial DNA has undergone during evolution have led to the inactivation or complete loss of a number of the 'native' transfer RNA genes deriving from the genome of the ancestral endosymbiont. Following sequence divergence in their genes, some native mitochondrial tRNAs are 'rescued' by editing, a post-transcriptional process which changes the RNA primary sequence. According to in vitro studies with the native mitochondrial tRNA(Phe) from potato and tRNA(His) from larch, editing is required for efficient processing. Some of the native tRNA genes which have been inactivated or lost have been replaced by tRNA genes present in plastid DNA sequences acquired by the mitochondrial genome during evolution, which raises the problem of the transcriptional regulation of tRNA genes in plant mitochondria. Finally, tRNAs for which no gene is present in the mitochondrial genome are imported from the cytosol. This process is highly specific for certain tRNAs, and it has been suggested that the cognate aminoacyl-tRNA synthetases may be responsible for this specificity. Indeed, a mutation which blocks recognition of the cytosolic Arabidopsis thaliana tRNA(Ala) by the corresponding alanyl-tRNA synthetase also prevents mitochondrial import of this tRNA in transgenic plants. Conversely, no significant mitochondrial co-import of the normally cytosol-specific tRNA(Asp) was detected in transgenic plants expressing the yeast cytosolic aspartyl-tRNA synthetase fused to a mitochondrial targeting sequence, suggesting that, although necessary, recognition by a cognate aminoacyl-tRNA synthetase might not be sufficient to allow tRNA import into plant mitochondria.

Biological Transport↗

The influence of antagonists of poly(ADP-ribose) metabolism on acetaminophen hepatotoxicity.

An array of therapeutically used analgetic and antirheumatic drugs causes severe liver damage. The present study investigates the hepatoprotective effects of inhibitors of NAD-dependent adenoribosylation reactions in analgesics-induced hepatic injury. Male NMRI mice were treated perorally with 500 mg/kg of acetaminophen, and the activities of both glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) were determined in serum. In addition, the activity of poly(ADP-ribose)polymerase (PARP) was quantified in liver cell nuclei. While the PARP-activity remained essentially unchanged, the acetaminophen-induced release of both GOT and GPT from injured liver cells could be inhibited by 90-99%, when mice were injected additionally with the selective PARP-inhibitors nicotinic acid amide, benzamide, caffeine, theophyline, and thymidine, respectively. We see the main application of inhibitors of adenoribosylation reactions as for the combinational use in pharmaceutical preparations of analgesics and antirheumatic drugs in order to avoid hepatic damage.

Acetaminophen↗