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1.
Arch Orthop Trauma Surg ; 135(10): 1343-7, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26188523

ABSTRACT

INTRODUCTION: The aim of the current study was to determine whether application of an intramedullary hip screw for definitive management of intertrochanteric fracture was associated with post-operative deformity. Specifically this study investigated whether nail insertion would cause a "wedge effect" of the intertrochanteric fracture manifesting as lateralization of the femoral shaft and varus malalignment. MATERIALS AND METHODS: The trauma database at the University of Pittsburgh Medical Center was investigated to identify all intertrochanteric fractures (AO/OTA 31A) over the past 3 years treated with an IMHS. Fractures eligible for inclusion were performed under the supervision of a fellowship trained orthopedic trauma surgeon. All fractures were reduced in optimal alignment using percutaneous or mini-open strategies during the reaming process and nail insertion. The entry portal was over-reamed by at least 1.5 mm. Cases selected for review of the "wedge effect" had optimal post-operative imaging allowing for assessment of discrepancy between the operative and normal hip. RESULTS: Forty six patients with an average age of 77 years were included for study. Fifty percent were classified as unstable patterns. Shaft lateralization following IMHS fixation of the fractured hip was found to be an average of 7 mm greater than the contralateral intact hip (p < 0.001) (range 0-30 mm). The neck-shaft angle of the operative hips was 129° as compared to 133° on the intact side (p = 0.009). The stability of the fracture pattern was not predictive for post-operative lateralization of the femoral shaft or varus angulation (p > 0.05) (Table 2). There was no difference in post-operative deformity among techniques used for maintenance of reduction during reaming and nail insertion (p > 0.05). Despite deformity, all cases demonstrated radiographic radiographic fracture union. CONCLUSION: Despite attention to detail, the application of an intramedullary hip screw for intertrochanteric fracture has the tendency to lateralize the shaft relative to the head/neck segment (The "wedge effect").


Subject(s)
Bone Screws , Femur/surgery , Fracture Fixation, Intramedullary/methods , Hip Fractures/surgery , Aged , Bone Nails , Female , Humans , Male , Treatment Outcome
2.
Scand J Immunol ; 81(3): 166-76, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25565108

ABSTRACT

The CC chemokine eotaxin contributes to epithelium-induced inflammation in airway diseases such as asthma. Eupatilin (5,7-dihydroxy-3',4',6'-trimethoxyflavone), a bioactive component of Artemisia asiatica Nakai (Asteraceae), is reported to inhibit the adhesion of eosinophils to bronchial epithelial cells. However, little is known about the molecular mechanism of eupatilin-induced attenuation of bronchial epithelium-induced inflammation. In this study, we investigated the effect of eupatilin on expression of eotaxin-1 (CCL11), a potent chemoattractant for eosinophils. Eupatilin significantly inhibited eotaxin expression in bronchial epithelial cells stimulated with TNF-α, while NF-κB and IκBα kinase (IKK) activities declined concurrently. Eupatilin also inhibited mitogen-activated protein kinase (MAPK) activity; however, all of these anti-inflammatory activities were reversed by MAPK overexpression. In contrast, eupatilin did not affect the signal transducer and activator of transcription 6 (STAT6) signalling in bronchial epithelial cells stimulated with IL-4. Furthermore, eupatilin significantly attenuated TNF-α-induced eosinophil migration. These results suggest that the eupatilin inhibits the signalling of MAPK, IKK, NF-κB and eotaxin-1 in bronchial epithelial cells, leading to inhibition of eosinophil migration.


Subject(s)
Chemokine CCL11/biosynthesis , Flavonoids/pharmacology , I-kappa B Kinase/antagonists & inhibitors , STAT6 Transcription Factor/drug effects , Transcription Factor RelA/antagonists & inhibitors , Asthma , Cell Adhesion/drug effects , Cell Line , Cell Movement/drug effects , Drugs, Chinese Herbal/pharmacology , Eosinophils/metabolism , Epithelial Cells/metabolism , Humans , I-kappa B Kinase/metabolism , Inflammation/immunology , Interleukin-4/pharmacology , MAP Kinase Signaling System/drug effects , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Respiratory Mucosa/metabolism , Transcription Factor RelA/metabolism , Tumor Necrosis Factor-alpha/pharmacology
3.
Drug Res (Stuttg) ; 64(10): 563-8, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24452518

ABSTRACT

To investigate the pathogenesis of hyperlipidemia-related erectile dysfunction and the effects of DA-8159, a new phosphodiesterase-5 inhibitor, on protein expression, we performed a proteomic analysis of differentially expressed proteins in the corpus cavernosum of hyperlipidemic rats by two-dimensional electrophoresis-mass spectrometry. Rats were fed high-cholesterol diet and treated with 5 mg·kg(-1)·day(-1) DA-8159 concurrently. After 5 months apparent hyperlipidemia and significantly decreased maximal intra-cavernous pressure were observed in the control group with the alteration of 8 proteins, which were restored by DA-8159 treatment. The proteins whose levels decreased >2-fold and attenuated by DA-8159 were determined alcohol dehydrogenase, aldolase A, annexin 1, and tropomyosin-rat, whereas proteins increased>2-fold and recovered by DA-8159 were found to be aldehyde dehydrogenase complex, guanine deaminase, creatine kinase-B, and phosphoglycerate mutase type B subunit.


Subject(s)
Erectile Dysfunction/metabolism , Hyperlipidemias/complications , Penis/metabolism , Proteins/metabolism , Proteomics , Animals , Biomarkers/metabolism , Disease Models, Animal , Electrophoresis, Gel, Two-Dimensional , Erectile Dysfunction/drug therapy , Erectile Dysfunction/etiology , Erectile Dysfunction/physiopathology , Male , Penile Erection , Penis/drug effects , Penis/physiopathology , Phosphodiesterase 5 Inhibitors/pharmacology , Proteomics/methods , Pyrimidines/pharmacology , Rats, Sprague-Dawley , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Sulfonamides/pharmacology , Time Factors
4.
Plant Biol (Stuttg) ; 15(2): 274-83, 2013 Mar.
Article in English | MEDLINE | ID: mdl-22726580

ABSTRACT

C3HC4-type RING zinc finger proteins are known to be essential in the regulation of plant processes, including responses to abiotic stress. Here, we identify, clone and examine the first C3HC4-type RING zinc finger protein (BrRZFP1) from Brassica rapa under stress conditions. Phylogenetic analysis of BrRZFP1 revealed strong sequence similarity to C3HC4-type zinc finger proteins from Arabidopsis that are induced by abiotic stresses. Diverse environmental stresses, including salt and cold, were found to induce BrRZFP1 transcripts greater than eightfold in B. rapa. Additional strong induction was shown of the stress hormone abscisic acid, together suggesting that BrRZFP1 could play a role as a general stress modulator. Similar profiles of induction for each of these stresses was found in both root and shoot tissues, although at much higher levels in roots. Constitutive expression of BrRZFP1 in Nicotiana tabacum was conducted to further analyse how changes in gene expression levels would affect plant stress responses. BrRZFP1 overexpression conferred increased tolerance to cold, salt and dehydration stresses. This was observed in several assays examining growth status throughout development, including increased germination, fresh weight and length of shoots and roots, as well as enhanced chlorophyll retention. These results suggest that the transcription factor BrRZFP1 is an important determinant of stress response in plants and that changes in its expression level in plants could increase stress tolerance.


Subject(s)
Brassica rapa/physiology , Cold Temperature , RING Finger Domains , Salt-Tolerant Plants/metabolism , Stress, Physiological , Abscisic Acid/pharmacology , Adaptation, Physiological , Agrobacterium tumefaciens/genetics , Agrobacterium tumefaciens/metabolism , Amino Acid Sequence , Base Sequence , Brassica rapa/genetics , Brassica rapa/metabolism , Chlorophyll/metabolism , Cloning, Molecular , Dehydration/metabolism , Gene Expression Regulation, Plant , Genes, Plant , Germination , Molecular Sequence Data , Phylogeny , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Roots/drug effects , Plant Roots/genetics , Plant Roots/metabolism , Plant Roots/physiology , Plant Shoots/drug effects , Plant Shoots/genetics , Plant Shoots/metabolism , Plant Shoots/physiology , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Plants, Genetically Modified/physiology , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Salt-Tolerant Plants/genetics , Salt-Tolerant Plants/physiology , Seeds/genetics , Seeds/metabolism , Seeds/physiology , Sodium Chloride/pharmacology , Nicotiana/genetics , Nicotiana/metabolism , Nicotiana/physiology , Transcription Factors/genetics , Transcription Factors/metabolism , Transcription, Genetic
5.
J Bone Joint Surg Br ; 94(12): 1632-6, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23188903

ABSTRACT

Intra-operative, peri-articular injection of local anaesthesia is an increasingly popular way of controlling pain following total knee replacement. At the same time, the problems associated with allogenic blood transfusion have led to interest in alternative methods for managing blood loss after total knee replacement, including the use of auto-transfusion of fluid from the patient's surgical drain. It is safe to combine peri-articular infiltration with auto-transfusion from the drain. We performed a randomised clinical trial to compare the concentration of local anaesthetic in the blood and in the fluid collected in the knee drain in patients having either a peri-articular injection or a femoral nerve block. Clinically relevant concentrations of local anaesthetic were found in the fluid from the drains of patients having peri-articular injections (4.92 µg/ml (sd 3.151)). However, none of the patients having femoral nerve blockade had detectable levels. None of the patients in either group had clinically relevant concentrations of local anaesthetic in their blood after re-transfusion. The evidence from this study suggests that it is safe to use peri-articular injection in combination with auto-transfusion of blood from peri-articular drains during knee replacement surgery.


Subject(s)
Anesthesia, Local/methods , Anesthetics, Local/blood , Arthroplasty, Replacement, Knee/methods , Blood Transfusion, Autologous/methods , Drainage/methods , Knee Joint/surgery , Nerve Block/methods , Aged , Anesthesia, Local/adverse effects , Anesthetics, Local/therapeutic use , Combined Modality Therapy , Female , Femoral Nerve , Humans , Injections, Intra-Articular , Male , Middle Aged , Treatment Outcome
6.
Scand J Immunol ; 74(3): 253-263, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21623862

ABSTRACT

DA-6034 is a synthetic derivative of eupatilin, a flavonoid with anti-inflammatory effects. The aim of this study was to investigate the effects of DA-6034 on the interactions between IκB kinase (IKK) and heat shock protein 90 (Hsp90), and activation of the nuclear factor-kappaB (NF-κB) signalling pathway in human gastric epithelial cells infected with Helicobacter pylori. MKN-45 gastric epithelial cell line was treated with DA-6034 and H. pylori. DA-6034 significantly inhibited NF-κB activation and upregulated the expressions of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 in MKN-45 cells infected with H. pylori. However, DA-6034 did not influence activator protein-1 DNA binding activity in H. pylori-infected gastric epithelial cells. Pretreatment with DA-6034 attenuated the H. pylori-induced increase in IKK activity, and Hsp90 was associated with IKK-α and IKK-γ in MKN-45 cells. Treatment with DA-6034 dissociated the Hsp90 and IKK-γ complex in H. pylori-infected cells, leading to the inhibition of IL-8 expression. These results suggest that the eupatilin derivative 7-carboxymethyloxy-3',4',5-trimethoxy flavone has anti-inflammatory activity in gastric epithelial cells infected with H. pylori through the promotion of the dissociation of the IKK-γ-Hsp90 complex and suppression of NF-κB signalling.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Flavonoids/pharmacology , Gastric Mucosa/microbiology , HSP90 Heat-Shock Proteins/metabolism , Helicobacter pylori/drug effects , I-kappa B Kinase/metabolism , NF-kappa B/metabolism , Anti-Inflammatory Agents, Non-Steroidal/metabolism , Cell Line , Chemokine CCL2/biosynthesis , Chemokine CCL2/genetics , Electrophoretic Mobility Shift Assay , Flavonoids/metabolism , Gastric Mucosa/metabolism , Helicobacter pylori/pathogenicity , Humans , Immunoblotting , Interleukin-8/biosynthesis , Interleukin-8/genetics , Polymerase Chain Reaction , Signal Transduction/drug effects , Transcription Factor AP-1/metabolism
7.
Asian J Androl ; 11(4): 435-42, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19465935

ABSTRACT

Erectile dysfunction (ED) is a major complication of diabetes mellitus (DM). This study investigates the relationship between ED and the downregulation of constitutive nitric oxide synthase (cNOS) in the corpus cavernosum (CC) of diabetic rats. It also examines the effects of udenafil, a phosphodiesterase type 5 (PDE5) inhibitor, on ED and cNOS expression levels. After 16 weeks of daily oral treatment with udenafil in diabetic rats, the intracavernous pressure/mean arterial pressure (ICP/MAP) ratio was recorded to measure erectile function, and cNOS expression was measured using reverse transcriptase (RT)-PCR and immunoblots. Although the ICP/MAP ratio and the expression levels of endothelial NOS (eNOS) and neuronal NOS (nNOS) in the CC were markedly decreased in diabetic rats, long-term udenafil treatment improved the erectile function and increased cNOS expression compared with diabetic controls. These findings suggest that ED in DM is closely related to decreased cNOS expression in the CC and that udenafil has the ability to compensate for this pathological change by modulating cNOS expression. Udenafil also has an inhibitory role in cGMP (cyclic guanosine monophosphate) degradation.


Subject(s)
Diabetes Mellitus, Experimental/enzymology , Gene Expression Regulation, Enzymologic/drug effects , Nitric Oxide Synthase Type I/metabolism , Penis/enzymology , Phosphodiesterase Inhibitors/pharmacology , Pyrimidines/pharmacology , Animals , Base Sequence , Blood Glucose/analysis , Body Weight/drug effects , DNA Primers , Diabetes Mellitus, Experimental/chemically induced , Disease Models, Animal , Electric Stimulation , Erectile Dysfunction , Male , Nitric Oxide Synthase Type I/genetics , Phosphodiesterase Inhibitors/administration & dosage , Polymerase Chain Reaction , Pyrimidines/administration & dosage , RNA, Messenger/genetics , Rats , Rats, Sprague-Dawley , Sulfonamides
8.
Int J Impot Res ; 17(5): 409-16, 2005.
Article in English | MEDLINE | ID: mdl-15920460

ABSTRACT

This study examined the effects of a new phosphodiesterase type 5 inhibitor, DA-8159, on erectile function associated with hypercholesterolemia. First of all, in order to investigate whether chronic administration of DA-8159 prevents the development of erectile dysfunction associated with hypercholesterolemia, male SD rats were divided into four groups (normal control, hypercholesterolemic control, DA-8159 5 or 20 mg/kg/day). Over a 5-month period, the animals were fed a 2% cholesterol diet and administered DA-8159 orally once a day. After 5 months, the electrostimulation-induced penile erection and the vascular function using acetylcholine-induced vasodilation with endothelium-intact aortic rings were examined. Furthermore, the plasma lipid profiles, endothelin and N(G),N(G)-dimethylarginine (asymmetrical dimethylarginine, ADMA) concentrations were measured. In order to investigate the acute treatment effect of DA-8159 on the erectile function in an established hypercholesterolemic model, additional animals were given a 2% cholesterol diet for 5 months without DA-8159. At the end of 5 months, the rats were divided into three groups (hypercholesterolemic control, DA-8159 0.3 or 1 mg/kg). DA-8159 was administered intravenously 1 min prior to the intracavernous pressure (ICP) measurement. In a chronic treatment study, while the hypercholesterolemic control showed a significantly lower erectile function, vascular reactivity, and increased plasma cholesterol, endothelin and ADMA concentration, the chronic DA-8159 treatment clearly restored the erectile responses by electric stimulation, preserved the potential of thoracic aortic relaxation in a dose-dependent manner, and significantly decreased the plasma endothelin and ADMA concentrations. In an acute treatment study, DA-8159 induced a dose- and frequency-dependent increase in ICP. The ICP/BP ratio and the corresponding AUC values, and the detumescence time were also significantly increased compared to the hypercholesterolemic control. These results suggest that DA-8159 is beneficial for erectile dysfunction in a rat hypercholesterolemic model and provided a rationale for the potential use of DA-8159 for treating erectile dysfunction secondary to hypercholesterolemia.


Subject(s)
3',5'-Cyclic-GMP Phosphodiesterases/antagonists & inhibitors , Erectile Dysfunction/drug therapy , Hypercholesterolemia/complications , Phosphodiesterase Inhibitors/pharmacology , Pyrimidines/pharmacology , Acetylcholine/pharmacology , Animals , Aorta/drug effects , Aorta/physiology , Arginine/analogs & derivatives , Arginine/blood , Cyclic Nucleotide Phosphodiesterases, Type 5 , Disease Models, Animal , Dose-Response Relationship, Drug , Endothelins/blood , Erectile Dysfunction/etiology , In Vitro Techniques , Lipids/blood , Male , Muscle Relaxation/drug effects , Penile Erection/drug effects , Penile Erection/physiology , Pressure , Rats , Rats, Sprague-Dawley , Sulfonamides
9.
Int J Impot Res ; 17(2): 134-41, 2005.
Article in English | MEDLINE | ID: mdl-15578039

ABSTRACT

The aim of this study was to assess the effect of phosphodiesterase 5 inhibitor, DA-8159, on erectile function throughout the quantitative analysis of vascular endothelial cell, smooth muscle (SM), TGF-beta1 expression in rat corpus cavernosum and measurement of intracavernous pressure (ICP) in diabetic rats. DA-8159 (0, 5, 10, 20 mg/kg) was administered orally once a day to diabetic rats. After 8 weeks, immunohistochemistry and computerized image analysis were performed to quantify the percent area within the Corpora Cavernosa occupied by the endothelial cells, SM cells and fibrotic tissues. ICP/mean arterial pressure (MAP) was also measured by electrostimulation of the cavernous nerve. Diabetic rats showed a significant decrease in the SM and endothelial cell content, and an increase in the TGF-beta1 expression level within the cavernosa areas compared to the normal rats. The mean cavernous SM, endothelial cell content and TGF-beta1 expression level were 9.7+/-0.7, 4.5+/-0.7 and 17.9+/-2.1%, respectively. DA-8159 prevented reduction of SM (12.3+/-0.4% (5 mg/kg), 13.8+/-0.4% (20 mg/kg)) and endothelial cell content (5.6+/-0.5% (5 mg/kg), 6.3+/-0.6% (20 mg/kg)). Immunoreactivity of TGF-beta1 and intracorporal fibrosis were also significantly lower in DA-8159-treated groups (11.8+/-1.2% (5 mg/kg), 9.5+/-1.1% (20 mg/kg)). Electrostimulation of the cavernous nerve induced significant increase in maximum ICP (62.2+/-13.6 mmHg in 10 mg/kg vs 37.5+/-17.5 mmHg in diabetic group) and area under the curve of the ratio of ICP/MAP (8891.09+/-1957 in 10 mg/kg vs 6315.87+/-2272 in diabetic group). These results suggest that subchronic treatment of DA-8159 can prevent the development of erectile dysfunction (ED), and provides a rationale for the use of DA-8159 as treatment of diabetic ED.


Subject(s)
3',5'-Cyclic-GMP Phosphodiesterases/antagonists & inhibitors , Diabetes Mellitus, Experimental/physiopathology , Penile Erection/physiology , Phosphodiesterase Inhibitors/pharmacology , Pyrimidines/pharmacology , Animals , Blood Pressure/drug effects , Cyclic Nucleotide Phosphodiesterases, Type 5 , Diabetes Mellitus, Experimental/pathology , Electric Stimulation , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Male , Muscle, Smooth/drug effects , Muscle, Smooth/metabolism , Penile Erection/drug effects , Penis/drug effects , Penis/pathology , Rats , Rats, Sprague-Dawley , Streptozocin , Sulfonamides , Transforming Growth Factor beta/drug effects , Transforming Growth Factor beta/metabolism , Transforming Growth Factor beta1
10.
Int J Impot Res ; 15(6): 405-11, 2003 Dec.
Article in English | MEDLINE | ID: mdl-14671658

ABSTRACT

DA-8159 is a pyrazolopyrimidinone derivative which exhibits potent and selective phosphodiesterase type 5 (PDE5) inhibition. The aim of this study was to investigate the effects of DA-8159 on inducing a penile erection in rabbits with an acute spinal cord injury (ASCI). DA-8159 was given either orally (1, 3, or 10 mg/kg) or intravenously (0.1 or 0.3 mg/kg) to conscious male albino rabbits with a surgical transection of the spinal cord at the L2-L4 lumbar vertebra or ischemic-reperfusion SCI rabbits. Erection was evaluated in a time-course manner by measuring the length of the uncovered penile mucosa. DA-8159 induced a dose-dependent erection in both transection and ischemic-reperfusion ASCI rabbits. The efficacy of DA-8159 was potentiated by an intravenous injection of sodium nitroprusside, a nitric oxide donor. Potentiation of the effect by nitric oxide donor implies that DA-8159 can enhance the erectile activity during sexual arousal. These results suggest that DA-8159 may be useful for treating erectile dysfunction in patients with an SCI.


Subject(s)
3',5'-Cyclic-GMP Phosphodiesterases/antagonists & inhibitors , Erectile Dysfunction/drug therapy , Penile Erection/drug effects , Pyrimidines/pharmacology , Spinal Cord Injuries/complications , Acute Disease , Animals , Area Under Curve , Cyclic Nucleotide Phosphodiesterases, Type 5 , Disease Models, Animal , Erectile Dysfunction/etiology , Male , Penile Erection/physiology , Rabbits , Reperfusion Injury/drug therapy , Spinal Cord/pathology , Spinal Cord Injuries/pathology , Sulfonamides
11.
J Int Med Res ; 31(6): 517-28, 2003.
Article in English | MEDLINE | ID: mdl-14708417

ABSTRACT

This study evaluated the effect of DA-8159, a new phosphodiesterase 5 inhibitor, on the compensatory development of right ventricular hypertrophy in monocrotaline (MCT)-induced pulmonary hypertension (PH). Rats treated with subcutaneous MCT were divided into three groups, which received DA-8159 1 mg/kg, DA-8159 5 mg/kg or saline-vehicle orally, twice daily for 21 days. The vehicle group demonstrated increased right ventricular weight, pulmonary artery medial wall thickening, myocardial fibrosis, increased plasma cyclic guanosine monophosphate (cGMP) concentration and reduced body weight gains. DA-8159, however, markedly attenuated the compensatory development of right ventricular hypertrophy and pulmonary artery medial wall thickening, amplified the increase in plasma cGMP levels and increased lung cGMP concentrations. In addition, DA-8159 prevented myocardial fibrosis induced by MCT. These results demonstrate that DA-8159 attenuates the compensatory development of right ventricular hypertrophy in a rate model of PH. DA-8159 might, therefore, be a useful treatment option for PH, but its efficacy in humans needs evaluating.


Subject(s)
Hypertension, Pulmonary/complications , Hypertrophy, Right Ventricular/prevention & control , Phosphodiesterase Inhibitors/pharmacology , Pyrimidines/pharmacology , 3',5'-Cyclic-GMP Phosphodiesterases , Animals , Body Weight/drug effects , Cyclic GMP/blood , Cyclic GMP/metabolism , Cyclic Nucleotide Phosphodiesterases, Type 5 , Disease Models, Animal , Heart Ventricles/anatomy & histology , Heart Ventricles/drug effects , Heart Ventricles/pathology , Hypertension, Pulmonary/chemically induced , Hypertension, Pulmonary/drug therapy , Hypertrophy, Right Ventricular/pathology , Lung/drug effects , Lung/metabolism , Male , Monocrotaline/adverse effects , Myocardium/pathology , Organ Size , Phosphoric Diester Hydrolases/drug effects , Pulmonary Artery/anatomy & histology , Pulmonary Artery/drug effects , Rats , Rats, Sprague-Dawley , Sulfonamides
12.
Mol Cells ; 12(2): 227-32, 2001 Oct 31.
Article in English | MEDLINE | ID: mdl-11710526

ABSTRACT

We developed a molecular method for the identification of the S-alleles of Brassicaceae, which belongs to the inbred line. This method is quicker and more precise than the existing methods. The genotype of the S-allele for 20 S-haplotypes of cabbage and 20 S-haplotypes of broccoli was determined by a pollination test. In order to identify the S-alleles, we performed PCR-RFLP with a mixture of the primers that are related to the S-locus glycoprotein (SLG) gene, which corresponds to the results of the pollination test. The selected primers amplified all of the single bands of about 1,150 bp in all 40 lines of cabbage and broccoli. Three out of 20 lines of cabbage were amplified by class I SLG specific primers, whereas all of the lines of the cabbage were amplified by class II SLG specific primers. Therefore, we could not classify class I and class II precisely by the class I and class II primers. However, 15 out of 20 lines of broccoli were amplified by the class I SLG specific primers. The remaining 5 lines were amplified with the class II SLG specific primers. We then digested the amplified PCR products with various restriction endonucleases and chose a restriction endonuclease, which accords exactly with the results of the diallel cross. The best one was HinfI. Its RFLP result was the same as that of the nucleotide sequence analysis. The 40 lines of cabbage and broccoli consisted of 16 different S-haplotypes. Therefore, the PCR-RFLP analysis was quicker and more precise in identifying the characteristics of S-haplotypes that are used in breeding. Also, we were able to check whether the lines could be mixed. The S-genotypes were difficult to determine due to the different flowering time.


Subject(s)
Brassica/genetics , Alleles , Amino Acid Sequence , Genes, Plant , Genotype , Glycoproteins/genetics , Inbreeding , Molecular Sequence Data , Plant Proteins/genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Sequence Homology, Amino Acid
13.
Arch Pharm Res ; 23(5): 471-6, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11059826

ABSTRACT

DA-8159, a new phosphodiesterase 5 inhibitor, was assessed for its erectogenic potential by a penile erection test in rats, the relaxation of isolated rabbit corpus cavernosum (CC), and estimation of the intracavernous pressure (ICP) in the anesthetized dog. Oral administration of DA-8159 (0.3 to 1 mg/kg) increased the number of erections in rats with increasing dosage, with the highest penile erection index at 10 mg/kg. DA-8159 induced the relaxation of phenylephrine (PHE)-induced contractions in the rabbit CC and decreased the IC50 of the nitric oxide donor sodium nitroprusside (SNP) in a dose-dependent fashion. In pentobarbital-anesthetized dogs, the intravenous administration of DA-8159 (1 approximately 300 g/kg) potentiated the increase in ICP induced by the intracavernosal SNP in a dose-related manner. These findings suggest that DA-8159 has significant therapeutic potential in the treatment of erectile dysfunction.


Subject(s)
Penile Erection/drug effects , Phosphodiesterase Inhibitors/pharmacology , Pyrimidines , Animals , Dogs , Dose-Response Relationship, Drug , In Vitro Techniques , Male , Nitroprusside/pharmacology , Rabbits , Rats , Rats, Sprague-Dawley , Sulfonamides
14.
Vet Hum Toxicol ; 42(4): 234-5, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10928692

ABSTRACT

A 1-mo toxicity study followed by a 1-mo recovery period of recombinant human basic fibroblast growth factor (bFGF) was performed using Beagle dogs at doses of 30, 120 or 480 mg/kg/d to estimate the no observed adverse effect level (NOAEL). Subcutaneous thickening was seen and its incidence, as well as that of stiffness of the injection sites, increased with dose. There were neither dead animals nor significant changes of body weight during the experimental period. In addition, no significant bFGF-related changes were found in ophthalmologic and histopathological examination, urinalysis and hematological, biochemical and organ weight parameters. At necropsy, red-brownish spots and/or nodule formations were recognized in a dose-dependent manner. Splenomegaly was noted in the 480 mg/kg group, but these findings had a low incidence in all dose groups. The findings in the dosing period disappeared or were ameliorated during the recovery period. The above data suggests the NOAEL of bFGF in Beagle dogs is >480 mg/kg/d.


Subject(s)
Carcinogens/toxicity , Dogs/metabolism , Fibroblast Growth Factor 2/toxicity , Recombinant Proteins/toxicity , Animals , Carcinogens/administration & dosage , Dogs/blood , Dogs/urine , Dose-Response Relationship, Drug , Drug Approval , Female , Fibroblast Growth Factor 2/administration & dosage , Guidelines as Topic , Humans , Injections, Subcutaneous/veterinary , Male , No-Observed-Adverse-Effect Level , Recombinant Proteins/administration & dosage , Splenomegaly/chemically induced
15.
Mol Cells ; 7(1): 136-9, 1997 Feb 28.
Article in English | MEDLINE | ID: mdl-9085279

ABSTRACT

To compare the gene order of the chloroplast genome among dicotyledonous plants, we constructed a physical map of chloroplast DNA (cpDNA) of Korean ginseng (Panax ginseng C.A. Meyer) with four restriction enzymes, BamHI, HindIII, EcoRI, and PstI. The restriction enzyme recognition sites of the physical map were also confirmed by Southern hybridization of total ginseng cpDNA with homologous and heterologous probes. The cpDNA of Korean ginseng was determined as a circular molecule with a total size of about 154 kb, which contain two inverted repeats of 23 kb each that disrupt the rest of the molecule into a large (90 kb) and a small single copy region (18 kb). The genome structure of Korean ginseng cpDNA was similar in size and gene order to that of tobacco cpDNA. The cpDNA of Korean and American ginseng (P. quinquefolius) showed very similar restriction patterns.


Subject(s)
Chromosome Mapping , DNA, Chloroplast/genetics , Panax/genetics , Plants, Medicinal , Cloning, Molecular , DNA, Circular/genetics , Genetic Variation , Genome, Plant , Korea , Repetitive Sequences, Nucleic Acid , Restriction Mapping , United States
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