Modulation of bone turnover by Cissus quadrangularis after ovariectomy in rats.

In women, age-related bone loss is associated with increased risk of bone fracture. Existing therapies are associated with severe side effects; thus, there is a need to find alternative medicines with less or optimal side effects. Cissus quadrangularis (CQ), an Ayurvedic medicine used to enhance fracture healing, was tested for its bone protective properties and studied to discern the mechanism by which it is beneficial to bone. Female Sprague Dawley rats were either sham operated or ovariectomized and were fed CQ for 3 months. Several biochemical markers, cytokines and hormones were assayed. Femur, tibia and lumbar vertebrae were subjected to pQCT and µCT densitometry. MC3T3 cells were cultured, treated with CQ and used to analyze miRNA content and subjected to qPCR for gene expression analysis related to bone metabolism. CQO rats showed protected bone mass and microarchitecture of trabecular bone in the distal femoral metaphysis and the proximal tibial metaphysis. The lumbar vertebrae, however, showed no significant changes. Serum protein expression levels of P1NP increased and Trap5b and CTX levels decreased with in vivo CQ treatment. Some influence on the anti- and pro-inflammatory markers was also observed. Significantly high level of estradiol in the CQO rats was observed. In vitro expression of a few genes related to bone metabolism showed that osteocalcin increased significantly. The other genes-collagen I expression, SPP1, BMP2, DCAT1-decreased significantly. Certain miRNA that regulate bone turnover using the BMP pathway and Wnt signaling pathways were upregulated by CQ. qPCR after acute treatment with CQ showed significantly increased levels of osteocalcin and decreased levels of Wnt/ß catenin antagonist DCAT1. Overall, CQ protected the microarchitecture of the long bones from ovariectomy-induced bone loss. This may be because of decreased inflammation and modulation through the BMP and Wnt signaling pathways. We conclude that CQ is a potential therapeutic agent to treat postmenopausal osteoporosis with no side effects.

Comparing DNA extraction methods for analysis of botanical materials found in anti-diabetic supplements.

A comparative performance evaluation of DNA extraction methods from anti-diabetic botanical supplements using various commercial kits was conducted, to determine which produces the best quality DNA suitable for PCR amplification, sequencing and species identification. All plant materials involved were of suboptimal quality showing various levels of degradation and therefore representing real conditions for testing herbal supplements. Eight different DNA extraction methods were used to isolate genomic DNA from 13 medicinal plant products. Two methods for evaluation, DNA concentration measurements that included absorbance ratios as well as PCR amplifiability, were used to determine quantity and quality of extracted DNA. We found that neither DNA concentrations nor commonly used UV absorbance ratio measurements at A(260)/A(280) between 1.7 and 1.9 are suitable for globally predicting PCR success in these plant samples, and that PCR amplifiablity itself was the best indicator of extracted product quality. However, our results suggest that A(260)/A(280) ratios below about 1.3 and above 2.3 indicated a DNA quality too poor to amplify. Therefore, A(260)/A(280) measurements are not useful to identify samples that likely will amplify but can be used to exclude samples that likely will not amplify reducing the cost for unnecessarily subjecting samples to PCR. The two Nucleospin(®) plant II kit extraction methods produced the most pure and amplifiable genomic DNA extracts. Our results suggest that there are clear, discernable differences between extraction methods for low quality plant samples in terms of producing contamination-free, high-quality genomic DNA to be used for further analysis.

Spermatotoxicity of a protein isolated from the root of Achyranthes aspera: a comparative study with gossypol.

BACKGROUND: A previous study showed that 50% ethanolic extracts of the roots of Achyranthes aspera possess spermatotoxic effects. STUDY DESIGN: A 58-kDa protein (Ap) was isolated, and its spermatotoxic effects were studied in comparison with gossypol. Ap (25 mg/kg body weight a day) and gossypol (40 mg/kg body weight a day) were administered orally to Swiss male albino mice for 35 days. Sperm motility, sperm count, sperm abnormality, toxicity markers such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) in the liver and serum, testicular activities of hydroxyl methyl glutaryl CoA reductase (HMG CoA reductase), 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase(17ß-HSD), glucose-6-phosphate dehydrogenase, cholesterol level and serum testosterone were assayed. Spermicidal action of the proteolytic digests of Ap was also studied in vitro. RESULTS: Treated mice showed significant spermatotoxicity. Significant differences were also observed in the testicular activities of HMG CoA reductase, 3ß-HSD, 17ß-HSD and glucose-6-phosphate dehydrogenase and in the levels of cholesterol and serum testosterone. The nontoxic nature of Ap was indicated by the insignificant alterations in the activities of AST and ALT. Ap possessed spermicidal activity even after proteolysis. CONCLUSION: The 58-kDa protein isolated from A. aspera possesses spermatotoxic effects comparable to gossypol.