14-Deoxy-11,12-didehydroandrographolide inhibits proliferation and induces GSH-dependent cell death of human promonocytic leukemic cells

14-Deoxy-11,12-didehydroandrographolide (AND2), an analogue of andrographolide, showed more potent cytotoxicity against human promonocytic leukemia (THP-1) cells than adherent cancer cell lines. In this study AND2 was isolated from the plant Andrographis paniculata and it was characterized. The antiproliferative effect of AND2 on both adherent (PC-3 and MDAMB) and non-adherent (THP-1 and Jurkat) cancer cell lines was evaluated by MTT assay. The effect of intracellular reduced glutathione (GSH) on AND2-induced cytotoxicity was studied by conducting cell viability assays on GSH-pretreated cells. The effect of AND2 on the redox status of THP-1 cells was determined by analyzing the endogenous reduced GSH content. Apoptosis induction was confirmed by DNA laddering assay and Western blot analysis using anti-caspase-3 protein antibody. AND2 showed antiproliferative action on both THP-1 and Jurkat cancer cell lines with low IC₅₀ values. Cytotoxicity of AND2 was reversed by GSH pretreatment. AND2 treatment decreased the GSH content by 19.76 % (p < 0.001) in the THP-1 cancer cell line and reduced the cell clumping between the THP-1 cells. Expression of procaspase-3 varied in THP-1 cells during the time course of AND2 treatment. Procaspase-3 expression reached a maximum in treated cells at 32 h and was markedly reduced at 48 h but no procaspase-3 cleavage was observed. The obtained results suggest that AND2 is more effective against leukemia cells. AND2 induced a redox-mediated cell death in THP-1 cells. As AND2 temporarily increased the procaspase-3 expression during treatment, this study encourages the preclinical testing of AND2 against promonocytic leukemia cells in combination with small molecules that directly activate procaspase-3 to caspase-3.     

Low Concentration of a Dioxin (2, 3, 7, 8 TCDD) Affects the Glycosidases and Acid Phosphatase Activity in Mice Hepatocytes

Present communication reports the effects of environmentally available, low doses of tetra chloro di benzo-p-dioxin (2,3,7,8 TCDD) to lysosomal enzymes in mice liver. The study tests the hypothesis, in vivo exposure of low dose TCDD provokes dose and duration dependent toxic effects to key lysosomal enzymes and thereby causes cellular apoptotic changes. Three groups of female Swiss albino mice were subjected to two doses of TCDD (0.004 mg/kg bw/d, 0.04 mg/kg bw/d) for 2, 4 and 6 days of exposure durations. The results indicated significant exposure duration dependent effects of TCDD in mice liver cells. The results suggested that TCDD possibly induced an increase in intracellular ions or ROS which in turn altered different physiological activities by affecting different metabolic pathway of the liver cells. The altered functions of key lysosomal enzymes by TCDD may also evoke the process of cellular apoptosis.     

Laparoscopic Ablative and Reconstructive Surgeries in Genitourinary Tuberculosis

Background and Objectives:

Laparoscopy is the present standard of care for urologic diseases. Laparoscopy in renal tuberculosis (genitourinary tuberculosis) is difficult because of inflammation and fibrosis associated with the disease. We present the outcome of our experience of laparoscopy in genitourinary tuberculosis, both ablative and reconstructive.

Methods:

The detailed data of patients with genitourinary tuberculosis who underwent laparoscopic surgeries between January 2011 and September 2012 were reviewed. Indications, type of surgery, duration, blood loss, intraoperative problems, postoperative outcomes, and follow-up details were noted.

Results:

Overall, 7 laparoscopic procedures were performed: 5 nephrectomies, 1 ureteric reimplantation with psoas hitch, and 1 combined nephrectomy and laparoscopy-assisted Mainz II pouch reconstruction. The mean operative time was 192 minutes for nephrectomy, 210 minutes for ureteric reimplantation, and 480 minutes for nephrectomy with Mainz II pouch reconstruction. There were no conversions to open surgery. The mean amount of blood loss was 70 mL for the nephrectomies, 100 mL for ureteric reimplantation, and 200 mL for nephrectomy with Mainz II pouch reconstruction. In 5 of 6 patients who underwent nephrectomy, there was severe perinephric and peripelvic fibrosis posing difficulty in dissection. However, the renal vessels could be controlled individually. The mean postoperative hospital stay was 3 days for the nephrectomies, 5 days for the ureteric reimplantation, and 10 days for the nephrectomy with Mainz II pouch reconstruction. In all cases the recovery was uneventful.

Conclusions:

Laparoscopy, though technically more demanding, is a feasible and safe option for ablative and complex reconstructive procedures in genitourinary tuberculosis. It offers the benefits of minimally invasive surgery. The difficulty with this procedure is mostly because of peripelvic and perinephric fibrosis, whereas the lower ureter and bladder are relatively easier to dissect.     

Root-associated fungi of Arabidopsis thaliana and Microthlaspi perfoliatum

Root-associated fungi, with the focus on endophytic species, were isolated from healthy Arabidopsis thaliana and Microthlaspi perfoliatum plants collected at different locations in Germany. A large number of fungal taxa were discovered with a small-scale approach. This provides additional evidence that root-associated and endophytic fungi are common in Brassicaceae. The most prevalent genera associated with A. thaliana roots were Trichoderma and Fusarium, while the roots of M. perfoliatum were dominated by different species of Fusarium and Penicillium. Differences in species composition and richness might be due to preferences and life-cycle of the two plant species. Strains of endophyte species that did not have closely related species in GenBank searches and those already known as root endophytes were chosen for preliminary co-cultivation experiments using germinating host plants on agar medium to observe effects on plant growth and health. Under these conditions several fungal isolates had an adverse effect on plant growth and health, especially on Arabidopsis thaliana. Some isolates did not adversely affect biomass during initial plant growth, while they altered the shoot-root ratio in favour of the shoot, especially in Microthlaspi perfoliatum. These strains are promising candidates for future research on endophytes as they might have some effects in Brassicaceae that are similar to mycorrhizal fungi. They are also promising candidates for investigating interactions with their host plants.