Biomedical subjects

C-S Lin

Publications and source records attributed to C-S Lin.

7 recordsLinked to original sources

PubMed2003

A method for inflorescence proliferation.

Most perennial plants must pass through a long juvenile phase of vegetative development before they are capable of flowering. We have developed a method specifying inflorescence proliferation to bypass juvenility and maintain the adult phase. Bamboo ( Bambusa edulis) inflorescences were amplified by incubation in Murashige and Skoog medium supplemented with 0.1 mg/l thidiazuron. Mutant albino inflorescences also proliferated in this medium. This method is equally effective with dicotyledonous plants. Ginseng ( Panax ginseng) buds were incubated in B5 medium supplemented with 1 mg/l benzyladenine and 1 mg/l gibberellic acid; new inflorescences developed from the base of the explants. Ginseng flowers were parthenocarpic and some of the fruit proliferated in vitro. Using the inflorescences as the material of somatic embryogenesis, we demonstrated that these were not mutations. The regenerated plants still had a juvenile phase and grew normally.

Bambusa↗

PubMed2003

Intracavernosal vascular endothelial growth factor (VEGF) injection and adeno-associated virus-mediated VEGF gene therapy prevent and reverse venogenic erectile dysfunction in rats.

Penile veno-occlusive dysfunction (venogenic erectile dysfunction) is a common cause of erectile dysfunction (ED). We investigated whether vascular endothelial growth factor (VEGF) can be used to prevent and reverse venogenic ED in a rat model. Pharmacological cavernosometry was developed and validated using adult male rats with either arteriogenic or venogenic ED. Castrated animals were treated with intracavernous VEGF as either a recombinant protein (C+VEGF) or adeno-associated virus (AAV)-mediated VEGF gene therapy (C+VEGF gene) in an attempt to prevent the development of venogenic ED. Other animal groups received testosterone replacement (C+testosterone) or intracavernous AAV-LacZ gene (C+LacZ). Animals with documented venogenic ED were treated with intracavernous VEGF in an attempt to reverse their ED. Functional analysis (pharmacological infusion cavernosometry) was performed following treatment. Penile specimens were harvested for immunohistochemistry and electron microscopic evaluation. Castrated rats showed a decrease in papaverine-induced intracavernous pressure and an increase in maintenance and drop rates during pharmacological cavernosometry. These changes were prevented by systemic testosterone and intracavernous VEGF or AAV-VEGF therapy. Moreover, intracavernous VEGF was able to reverse the venogenic ED produced by castration. The quantity of penile smooth muscle detected by alpha actin staining decreased after castration but not in the C+T, C+VEGF, or C+VEGF gene groups. Transmission electron microscopy revealed atrophy of penile smooth muscle cells and nerves in the castrated rats. In VEGF-treated rats, regeneration of smooth muscle and nerves as well as endothelial cell hypertrophy and hyperplasia were the prominent features. In our animal model, systemic testosterone replacement or intracavernous VEGF (protein and VEGF gene) prevented the veno-occlusive dysfunction in castrated animals. In rats with established venous leakage, VEGF treatment reversed the cavernosometric findings of leakage. Intracavernous injection of either VEGF protein or VEGF gene may be a preferred therapy to preserve erectile function in patients in whom testosterone therapy is contraindicated.

Adenoviridae↗

PubMed2003

Isolation of two isoforms of phosphodiesterase 5 from rat penis.

Inhibition of cGMP-specific phosphodiesterase type 5 (PDE5) has been shown to improve penile erection in patients with erectile dysfunction. We have reported previously the cloning of three PDE5 isoforms from human penile tissues. Here we report the cloning of two PDE5 isoforms from rat penile tissues. The similarity between rat and human PDE5A1-specific sequences were 68 and 88% at the nucleotide and amino-acid levels, respectively. Like the bovine and canine PDE5A1 sequences, the rat PDE5A1 sequence lacks the polyglutamine tract that appears to be unique to the human PDE5A1 sequence. The similarity between rat and human PDE5A2-specific sequences were 64 and 100% at the nucleotide and amino-acid levels, respectively. The equivalent of human PDE5A3-specific sequence was identified in the rat PDE5A gene; however, repeated efforts to clone the putative rat PDE5A3 isoform were not successful. Expression of PDE5A1 and A2 mRNA in various tissues was examined by Northern blotting and reverse transcription-polymerase chain reaction. Results from the two experimental procedures were largely in good agreement and indicated that PDE5A1 and A2 mRNA were expressed in a tissue-specific manner with PDE5A2 being the dominant isoform. International Journal of Impotence Research (2003) 15, 129-136. doi:10.1038/sj.ijir.3900983

3',5'-Cyclic-GMP Phosphodiesterases↗

PubMed2002

Human PDE5A gene encodes three PDE5 isoforms from two alternate promoters.

Sildenafil improves erectile function by inhibiting the cGMP-catalytic activity of phosphodiesterase type V (PDE5). We used rapid amplification of cDNA Ends-polymerase chain reaction (RACE-PCR) to isolate three PDE5 isoforms from human corpus cavernosum. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis on eight human cavernous tissue samples showed that all samples expressed the PDE5A1 at a lower level than the PDE5A2 isoform. Five samples expressed the PDE5A3 isoform at various levels while the other three did not. Analysis on non-penile tissues showed that all tissues expressed the A1 and A2 isoforms while only those that have substantial amounts of smooth muscle expressed the A3 isoform. Cloning and sequencing of the PDE5A gene showed that the isoform-specific 5'-ends of the PDE5 mRNAs are encoded from three alternative first exons arranged in the order of A1-A3-A2. Promoter activities were detected upstream from the A1-specific exon and in the intron preceding the A2-specific exon. The upstream PDE5A promoter is expected to direct the expression of all three PDE5 isoforms while the intronic PDE5A2 promoter only the A2 isoform. Both promoters were upregulated by increasing concentrations of either cAMP or cGMP. Several transcription factor AP2 and Sp1-binding sequences identified in the promoters are likely to be the mediators of cAMP/cGMP-responsiveness.

3',5'-Cyclic-GMP Phosphodiesterases↗

PubMed2002

Cyclic AMP and cyclic GMP activate protein kinase G in cavernosal smooth muscle cells: old age is a negative factor.

OBJECTIVE: To investigate protein kinase G-I (PKG-I) expression and activation in cavernosal smooth muscle cells (CSMC) of young and old rats. MATERIALS AND METHODS: PKG-I expression in rat penis was examined by immunohistochemical staining, reverse transcription-polymerase chain reaction, and Western blot analysis. CSMC isolated from young (16-week-old) and old (28-month-old) rats were grown as monolayer cell cultures and treated with different dosages of cAMP or cGMP for different periods. Their proteins were then analysed for the expression of vasodilator-stimulated phosphoprotein (VASP), phosphorylated VASP (at serine 239), PKG-I, and protein kinase A (PKA). RESULTS: PKG-I expression was detected in the vascular and CSMC of the rat penis. There was little or no difference in the level of PKG-I expression between young and old rats. Treatment of CSMC with different dosages of cAMP or cGMP did not change the expression levels of VASP throughout the entire test period (up to 24 h). In contrast, the level of VASP phosphorylation at S239, i.e. the level of PKG-I activation, depended on the dosages of cAMP and cGMP and on the duration of treatment. Prolonged treatment (24 h) with either cAMP or cGMP resulted in down-regulation of both PKG-I and PKA. While cAMP and cGMP produced very similar results in nearly every aspect, there was a difference in one test, in which cGMP produced much less activated PKG-I than cAMP in the CSMC of 28-month-old-rats. CONCLUSIONS: For the first time we provide evidence for PKG-I activation in CSMC. Both cAMP and cGMP were capable of activating PKG-I in CSMC. Age seemed to compromise the ability of PKG-I in response to cGMP.

Age Factors↗

PubMed2002

Intracavernosal injection of vascular endothelial growth factor induces nitric oxide synthase isoforms.

OBJECTIVE: To identify genes that are affected by vascular endothelial growth factor (VEGF), as an intracavernosal injection with VEGF improved the recovery of erectile function in a rat model of arteriogenic impotence, specifically examining the three nitric oxide synthase (NOS) genes, nNOS, eNOS, and iNOS. MATERIALS AND METHODS: Male rats had their pudendal arteries ligated or underwent a sham operation. They were then treated by an intracavernosal injection with 4 microg of VEGF in phosphate-buffered saline (PBS) or PBS alone. At 6 and 24 h after treatment electrostimulation was applied to the cavernosal nerve and the intracorporal pressure measured. The erectile tissue was then harvested for RNA isolation and cryo-sectioning. The isolated RNA was used for microarray and reverse transcription-polymerase chain reaction (RT-PCR) analyses, and the tissue sections for immunohistochemical analysis. RESULTS: Microarray analysis detected nNOS, eNOS and iNOS at very low expression levels in PBS-treated rats; expression levels were higher for eNOS and iNOS in all VEGF-treated rats. These results were further confirmed by RT-PCR analysis. Immunohistochemical analysis identified the cavernosal endothelium and smooth muscle as the tissue types where eNOS and iNOS were up-regulated, respectively. CONCLUSIONS: This is the first report of the induction of both eNOS and iNOS in the penis after intracavernosal VEGF. These events may help support a significant recovery of erectile function after interrupting the blood supply to the penis.

Animals↗

PubMedDate not supplied

Complete atrioventricular block following myocarditis in children.

Complete atrioventricular block (CAVB) can be either congenital or acquired in children. Acquired CAVB is occasionally seen in myocarditis patients. To determine the etiology, natural history, and outcome of children with acquired nonsurgical CAVB, we retrospectively reviewed nine children who had suffered CAVB caused by suspected infectious myocarditis. All of them had CAVB with a wide QRS escape ventricular rhythm on admission. Three of them had ventricular tachycardia in addition to CAVB. Seven of them had a preceding upper respiratory tract infection. All of them had congestive heart failure. Five of them had Stokes-Adams seizures. Three etiologies were identified in four of the children. All patients received inotropic agents and emergency temporary pacing. In all except one case, the cardiac rhythm returned to sinus rhythm within 10 days. During a follow-up period of 9 to 96 months, all were asymptomatic and drug-free. Electrocardiograms showed that four patients were completely normal, there was complete RBBB in four and left anterior fascicular block in one patient. We conclude that although CAVB associated with myocarditis can be life-threatening, the long-term prognosis is good if patients are diagnosed early and proper management is employed.

Adolescent↗