ABSTRACT
No Abstract Keywords.
Subject(s)
Anti-Arrhythmia Agents , Calcium , Lipids , Verapamil , Anti-Arrhythmia Agents/adverse effects , Calcium Channel Blockers , Fat Emulsions, Intravenous , Humans , Verapamil/adverse effectsABSTRACT
This study aimed to examine the effects of lipid emulsion on the vasodilation and cardiovascular depression induced by toxic doses of calcium channel blockers. The effects of lipid emulsion on the vasodilation induced by bepridil, verapamil, nifedipine, and diltiazem were investigated in isolated endothelium-denuded rat aortae. The effect of lipid emulsion on the comparable hemodynamic depression induced by the continuous infusion of a toxic dose of either verapamil or diltiazem was examined in an in vivo rat model. The results showed the following decreasing order for the magnitude of lipid emulsion-mediated inhibition of vasodilation: bepridil, verapamil, nifedipine, and diltiazem. Lipid emulsion (0.5-2%) reversed the vasodilation induced by a toxic dose of verapamil, whereas only a higher concentration (2%) reversed the vasodilation induced by a toxic dose of diltiazem. Pretreatment with lipid emulsion alleviated the systolic and mean blood pressure decreases induced by a toxic dose of verapamil, whereas it had no effect on the decrease induced by diltiazem. Taken together, these results suggest that lipid emulsion alleviates the severe vasodilation and systolic blood pressure decrease induced by a toxic dose of verapamil, and this alleviation appears to be associated with the relatively high lipid solubility of verapamil.
Subject(s)
Blood Pressure/drug effects , Calcium Channel Blockers/toxicity , Phospholipids/therapeutic use , Soybean Oil/therapeutic use , Vasodilation/drug effects , Vasodilator Agents/toxicity , Verapamil/toxicity , Animals , Aorta, Thoracic/drug effects , Aorta, Thoracic/physiology , Bepridil/toxicity , Diltiazem/toxicity , Emulsions/pharmacology , Emulsions/therapeutic use , In Vitro Techniques , Male , Nifedipine/toxicity , Phenylephrine/pharmacology , Phospholipids/pharmacology , Rats , Rats, Sprague-Dawley , Soybean Oil/pharmacologyABSTRACT
The goal of this in vitro study was to investigate the effect of lipid emulsion on apoptosis induced by a toxic dose of bupivacaine (BPV) in H9c2 rat cardiomyoblast cell lines. The effect of lipid emulsion on the decreased cell viability and count induced by BPV or mepivacaine (MPV) in the H9c2 cells was assessed using an 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay or a cell count assay. The effect of BPV or lipid emulsion combined with BPV on cleaved caspase 3, caspase 8, and Bax in H9c2 cells was investigated using Western blotting. A terminal deoxynucleotidyl transferase dUTP2'-deoxyuridine 5'-triphosphate nick end-labeling (TUNEL) assay was performed to detect apoptosis of H9c2 cells treated with BPV alone or lipid emulsion combined with BPV. The magnitude of lipid emulsion-mediated attenuation of decreased cell viability induced by BPV was higher than that of lipid emulsion-mediated attenuation of decreased cell viability induced by MPV. Lipid emulsion attenuated the increases in cleaved caspase 3, caspase 8 and Bax induced by BPV. Lipid emulsion attenuated the increases in TUNEL-positive cells induced by BPV. These results suggest that lipid emulsion attenuates a toxic dose of BPV-induced apoptosis via inhibition of the extrinsic and intrinsic apoptotic pathways. The protective effect of lipid emulsion may be partially associated with the relatively high lipid solubility of BPV.
Subject(s)
Anesthetics, Local/toxicity , Apoptosis/drug effects , Bupivacaine/toxicity , Lipids/pharmacology , Myoblasts, Cardiac/drug effects , Anesthetics, Local/administration & dosage , Animals , Bupivacaine/administration & dosage , Cardiotoxicity/prevention & control , Cell Line , Cell Survival/drug effects , Dose-Response Relationship, Drug , Emulsions , In Situ Nick-End Labeling , Lipids/administration & dosage , Lipids/chemistry , Myoblasts, Cardiac/pathology , RatsSubject(s)
Barbiturates/adverse effects , Coma/chemically induced , Craniocerebral Trauma/therapy , Hypokalemia/chemically induced , Accidents, Traffic , Adult , Barbiturates/therapeutic use , Cerebral Hemorrhage, Traumatic/surgery , Craniotomy , Female , Humans , Potassium/blood , Water-Electrolyte Balance/physiologyABSTRACT
A subtracted library was constructed from planthopper-infested wild rice (Oryza minuta) by suppression subtractive hybridization in combination with mirror orientation selection. To screen the differentially expressed transcripts in the library, we applied a cDNA microarray containing 960 random clones in a reverse Northern blot analysis using cDNA probes prepared from the mRNAs of control and planthopper-infested samples. On the basis of the signal intensities and expression ratios obtained from experiments performed in triplicate, we selected 383 clones. The elevated expression levels and overall profiles over time were verified by Northern blot analysis. Although Southern blot analysis showed similar copy numbers of the screened genes in O. minuta and O. sativa, it also revealed that the expression profiles had a different pattern. Functional categorization placed the identified transcripts in the categories of subcellular localization, metabolism, and protein fate. The presence of these expressed sequence tags implies that resistance of O. minuta to insect infestation can be achieved not only by an elevated expression of defense-related genes but also by enhanced metabolic activities.
Subject(s)
Gene Expression Regulation, Plant/physiology , Insecta/physiology , Oryza/genetics , Animals , DNA, Plant , Expressed Sequence Tags , Gene Library , Microarray Analysis , Oryza/parasitologyABSTRACT
The expressed sequence tags (ESTs) presented in this report are the first transcriptomes of wild rice. A cDNA library was constructed from 4-week-old leaf samples of greenhouse-grown Oryza minuta. The 5,211 cDNA clones of O. minuta represent 3,401 unique sequences, consisting of 2,787 singletons and 614 assembled sequences. Database comparisons of the cDNAs in GenBank's non-redundant databases using BLAST revealed that 4,957 of the 5,211 cDNAs (95.1%) showed a high degree of sequence homology to genes from other organisms. Most of the transcripts identified were genes related to metabolism, energy, protein biosynthesis and subcellular localization. The metabolism and energy categories of the O. minuta ESTs showed a considerably higher gene expression level than those of O. sativa ESTs. These data and genes can be utilized in rice breeding.
Subject(s)
Expressed Sequence Tags , Oryza/genetics , Plant Leaves/genetics , Arabidopsis/genetics , Base Sequence , Gene Expression Profiling , Gene Library , Molecular Sequence Data , Sequence Analysis, DNAABSTRACT
To identify fungal stress-related genes in wild rice, Oryza minuta, we constructed a subtracted library using suppression subtractive hybridization in combination with mirror orientation selection. DNA chips containing 960 randomly selected cDNA clones were applied by reverse Northern analysis to eliminate false positive clones from the library and to prescreen differentially expressed genes. In total, 377 cDNA clones were selected on the basis of their signal intensities and expression ratios. Sequence analyses of these 377 cDNA fragments revealed that 180 of them (47.7%) represented unique genes. Of these 180 cDNAs, 89 clones (49.6%) showed significant homologies to previously known genes, while the remaining 91 did not match any known sequences. The putative functions of the 180 unique ESTs were categorized by aligning them with MIPS data. They were classified into seven different groups using microarray data-derived expression patterns and verified by Northern blotting.
Subject(s)
Genes, Plant/physiology , Magnaporthe/genetics , Oryza/genetics , Oryza/microbiology , Blotting, Northern/methods , DNA, Fungal/genetics , DNA, Plant/genetics , Expressed Sequence Tags , Gene Expression Profiling , Gene Library , Magnaporthe/pathogenicity , Oligonucleotide Array Sequence Analysis , Sequence Alignment , Sequence Analysis, DNAABSTRACT
The suppression subtractive hybridization (SSH) method was used to isolate developmentally regulated genes during carnation flower maturation. Carnation flower maturation-related clones obtained by the SSH were serially assigned as CFMI (carnation flower maturation-induced) clones. Northern blot analysis showed that several CFMI clones were differentially expressed during flower development. One of the clones, CFMI-3, showed similarity to various animal secretory phospholipases A2 (PLA2). Since little is known about PLA2 gene sequence in plant species, the CFMI-3 clone was selected for further characterization by sequence analysis. Full sequence analysis reveals that the CFMI-3 contains a Ca2+ binding domain, a PLA2 active site, and 12 conserved Cys residues, which is a distinct characteristic of PLA2. Amino acid sequence alignment of CFMI-3 to various putative plant PLA2 confirmed that the CFMI-3 cDNA is the full-length putative PLA2 cDNA identified in plant species.
