Vascular rarefaction in peripheral skeletal muscle after experimental heart failure

A decrease in vascular density in peripheral skeletal muscle has been associated with exercise intolerance in humans with congestive heart failure (CHF). The purpose of this study was to determine whether CHF results in a reduction in vascular density in peripheral skeletal muscle. In this established model, CHF was induced by coronary artery ligation in New Zealand White rabbits and sham rabbits that underwent an identical surgical procedure without ligation of the coronary artery. At study termination, rabbits underwent hemodynamic testing and skeletal muscle analysis. The first series of rabbits was divided into sham (n = 6) and CHF (n = 6) 21 days postoperatively. Ten CHF rabbits were then examined 3 (n = 3), 7 (n = 3), and 14 days (n = 4) postoperatively. Vascular density in sham tibialis anterior muscle was 347 +/- 41 capillaries/mm2 or 1.20 +/- 0.11 capillaries/muscle fiber. In 21-day CHF rabbits, the capillary density was significantly lower, 236 +/- 14 capillaries/mm2 or 0.84 +/- 0.04 capillaries/muscle fiber (both P < 0.00001 vs. sham); PECAM protein was 2-fold lower (P < 0.0001) in muscle protein lysates; the fraction of apoptotic cells was greater, 3.8 +/- 2.2 vs. 0.69 +/- 0.56 (P < 0.02 vs. sham) with many TdT-mediated dUTP-biotin nick-end labeling-positive endothelial cells; and Bax protein was 2.8-fold greater (P < 0.0001). By regression analysis, vascular density tended to decrease over time (r2 = 0.572, P < 0.0001). Vascular rarefaction and endothelial apoptosis develop after experimental CHF and may contribute to the skeletal muscle abnormalities in this disease. Modulating vascular density may provide new approaches to treat exercise intolerance in CHF.     

Catheter-mediated subselective intracoronary gene delivery to the rabbit heart: introduction of a novel method

BACKGROUND: Recent studies suggest that gene therapy using replication-deficient adenoviruses will benefit treatment of cardiovascular diseases including heart failure. A persistent hurdle is the effective and reproducible delivery of a transgene to the myocardium with minimal iatrogenic morbidity. In this study, we sought to design a relatively non-invasive percutaneous gene delivery system that would maximize cardiac transgene expression and minimize mortality after intracoronary adenovirus injection. METHODS: Adult rabbits received a left circumflex coronary artery (LCx) infusion of 5x10(11) total viral particles of an adenovirus containing the marker transgene beta-galactosidase (Adeno-betaGal) via either a continuous infusion method utilizing an oxygenated, normothermic, physiologic pH Krebs solution driven by a Langendorff apparatus (n=12) or a timed bolus and set concentration at a constant infusion rate to the LCx (n=12). Six rabbits underwent global transgene delivery via an invasive method involving intraventricular delivery and aortic root cross-clamping. The efficacy of transgene expression via these three distinct delivery methods was determined in the left ventricle at 5 days by histological staining and colorimetric quantification assay. RESULTS: While the open-chest, aortic cross-clamping method provides the highest level of gene expression throughout the heart, the morbidity of this procedure is clinically prohibitive. Percutaneous LCx delivery of Adeno-betaGal using the Langendorff apparatus was associated with the lowest morbidity and mortality while still supporting significant myocardial gene expression. CONCLUSIONS: Percutaneous delivery of an adenovirus solution using a continuous infusion of oxygenated Krebs solution via a Langendorff apparatus appears to be a gene delivery modality offering the best compromise of gene expression and clinical utility to maximize any potential therapeutic outcome.     

Phytoremediation of lead-contaminated soil by Sinapis arvensis and Rapistrum rugosum

Nowadays, public concern relating to ecological deleterious effects of heavy metals is on the rise. To evaluate the potential of Rapistrum rugosum and Sinapis arvensis in lead- contaminate phytoremediate, a pot culture experiment was conducted. The pots were filled by soil treated with different rates of leadoxide (PbO) including 0 (control), 100, 200, 300, 400, and 500 mg Pb per 1 kg soil. Germinated seeds were sown. Surprisingly, with increasing concentration of Pb, dry weight of R. rugosum and S. arvensis did not decrease significantly. In both of species, the concentration of Pb was higher in roots than shoots. In general, S.arvensis was absorbed more Pb compared to R. rugosum. The results revealed high potential of R. rugosum and S. arvensis in withdrawing Pb from contaminated soil. For both species, a positive linear relation was observed between Pb concentration in soil and roots. However, linear relationship was not observed between Pb concentration in the soil and shoots. Although both species test had low ability in translocation Pb from roots to shoots but they showed high ability in uptake soil Pb by roots. Apparently, these plants are proper species for using in phytoremediation technology.     

Long term organ culture of rat liver rudiment in a synthetic medium: morphological and biochemical development.

Embryonic rat liver rudiments were grown and maintained in a chemically defined medium for up to 16 weeks. Liver explants grew and developed liver plates, bile canaliculi and ductules. Liver cells differentiated and, by the end of the second culture week, revealed cytological and ultrastructural features of adult rat liver cells. Glycogen synthetase activity appeared and reached a plateau within 2 weeks.Plasma proteins produced by the explants appeared sequentially in the culture medium. Their relative concentrations in the medium increased progressively to a plateau and they persisted throughout the culture period. Albumin was present in trace amounts during early days of culture. Its relative concentration increased and it became the major protein constituent of the medium by the second week.Transferrin, α₁M-globulin, haptoglobin, β₁c- and α₁B-globulin appeared in that order. The three fibrinogen subunits were identified during the first week of culture.Fetal proteins were identified during the early culture period, the most prominent being α-fetoprotein. The relative concentration of fetal proteins declined steadily as the culture progressed.     

Pancreas acinar cell differentiation VIII. Effect of methionine on DNA synthesis of pancreas anlage in organ culture

Rat pancreas embryonic rudiments incubated in a control chemically-defined (CD) medium grew and differentiated during 9 days of organ culture to adult acinar cells characterized by zymogen granules and relatively high levels of enzyme activity. Incubation of the anlagen in a chemically-defined medium with a lower methionine concentration (CD-MD) gave growth to a comparable size but no acinar cell formation, no zymogen granules and no increases of enzyme activity. Anlagen incubated in the CD medium showed a marked decrease of the autoradio-graphic labeling index of acinar cell nuclei after exposure to ³H-thymidine during the period of organ culture, whereas culture of anlagen in the CD-MD medium led to far less decrease of the acinar cell labeling index. When anlagen which had been grown in the CD-MD medium for 9 days were incubated for 3 additional days in the CD medium there was a highly significant drop in the previously high labeling index. The higher level of methionine in the CD medium may have been involved in the synthesis of, or the methylation of, an inhibitor of DNA synthesis.     

Nasal augmentation with split calvarial grafts in Orientals

This study reports on my experience with autogenous split calvarial grafts in nasal augmentation in 62 Orientals. In 78 percent of patients, the procedure was performed under local anesthesia in an outpatient setting. Total operating time for harvesting of split calvarial grafts ranged from 20 to 55 minutes, with a mean of 32 minutes. Patients ranged in age from 16 to 48 years, with a mean of 27 years. Follow-up was from 6 months to 8 years, with an average of 3.1 years. Intraoperative discomfort was uniformly low and well tolerated when local anesthesia was used. The complication rate was 8.0 percent, with three cases of minor seroma-hematoma formation at the bone-graft donor site. These were treated with aspiration. There were two recipient-site complications, with one case of complete bone resorption that occurred in a densely fibrotic nose with preexisting septal perforation and a case of overcorrection that was successfully rasped 1 year later. Because of their easy accessibility beneath the scalp, split calvarial grafts to the nose are useful in various types of nasal augmentation, and the technique is offered as a practical alternative to the use of alloplastic materials.     

Caspase-dependent apoptosis induced by two synthetic halogenated flavanones, 3′,7-dichloroflavanone and 3′,6-dichloroflavanone,on human breast and prostate cancer cells

The destruction of cancer cells with chemotherapeutic agents is normally achieved through apoptosis. We previously introduced two synthetic halogenated flavanone derivatives, 3^′,7-dichloroflavanone (3′-7 DCF) and 3^′,6-dichloroflavanone (3′-6 DCF), as potential apoptosis-inducing agents. In the current study, we investigated the ability of these compounds in triggering intrinsic or/and extrinsic pathway of apoptosis in breast and prostate cancer cells. Also, the synergistic effect of 3′-7 DCF with TLR3 (Toll-like receptor 3) agonist in apoptosis induction was evaluated on PC3 and LNCaP human prostate cancer cells. The involved pathway of apoptosis in the treated cells was delineated by caspase-3 activity assay, PARP-1 (poly(ADP-ribose)polymerase-1) cleavage, and procaspase-9 cleavage as markers of the intrinsic pathway and procaspase-8 cleavage as the marker of the extrinsic pathway. With the exception of the normal cells, treatment of all cell lines with both 3′-7 DCF and 3′-6 DCF triggered the cleavage of procaspase-8 and procaspase-9. These results indicate that the intrinsic and the extrinsic pathways of apoptosis are the mechanisms of the toxicity of flavanones in these cancer cell lines. However, the cytoxicity of the compound 3′-7 DCF was not synergistic with TLR3 agonist. Interestingly, the activation of caspases-9 preceeded that of caspase-8 suggesting that the intrinsic pathway is the primary reason for apoptosis induction by the flavanones.