SG Treatment Research Articles

Suicide plus immune gene therapy prevents post-surgical local relapse and increases overall survival in an aggressive mouse melanoma setting.

In an aggressive B16-F10 murine melanoma model, we evaluated the effectiveness and antitumor mechanisms triggered by a surgery adjuvant treatment that combined a local suicide gene therapy (SG) with a subcutaneous genetic vaccine (Vx) composed of B16-F10 cell extracts and lipoplexes carrying the genes of human interleukin-2 and murine granulocyte and macrophage colony stimulating factor. Pre-surgical SG treatment, neither alone nor combined with Vx was able to slow down the fast evolution of this tumor. After surgery, both SG and SG + Vx treatments, significantly prevented (in 50% of mice) or delayed (in the remaining 50%) post-surgical recurrence, as well as significantly prolonged recurrence-free (SG and SG + Vx) and overall median survival (SG + Vx). The treatment induced the generation of a pseudocapsule wrapping and separating the tumor from surrounding host tissue. Both, SG and the subcutaneous Vx, induced this envelope that was absent in the control group. On the other hand, PET scan imaging of the SG + Vx group suggested the development of an effective systemic immunostimulation that enhanced (18)FDG accrual in the thymus, spleen and vertebral column. When combined with surgery, direct intralesional injection of suicide gene plus distal subcutaneous genetic vaccine displayed efficacy and systemic antitumor immune response without host toxicity. This suggests the potential value of the assayed approach for clinical purposes.

Grazing Management of Flaccidgrass Pastures in Autumn

ABSTRACT‘Carostan’ flaccidgrass (Pennisetum flaccidum Griseb.) is a perennial warm‐season grass with potential for pasture, but appropriate autumn (September to November) management is needed to maintain forage mass (FM) production and prevent declines in plant density. We evaluated 10 treatments over 3 yr consisting of an ungrazed control, four treatments with multiple grazing events (MG), four treatments with a single grazing event (SG), and one treatment grazed after frost. The winter residues after the treatments were greatest (P < 0.01) when ungrazed compared with the grazed treatments (3347 vs. 2265 kg ha−1) and differed (P ≤ 0.01) among and between (P < 0.01) the MG and SG treatments. Plant density decreased from 13 to 27% with 8 to 10 wk of regrowth before frost. Greater plant density decline (60%) occurred when plots were grazed three times and only allowed 4 wk of regrowth before frost. Total nonstructural carbohydrates (TNC) of ungrazed basal stems, crowns, and roots sampled in December were 158, 193, and 250 g kg−1, respectively, and when grazed were 124, 183, and 253 g kg−1, respectively. By April TNC had declined 56, 35, and 25%, respectively. Plant density was not related to autumn total FM, FM regrowth, or overwintering residue but was related (P ≤ 0.02) to TNC of December roots (r = 0.59) and TNC of April basal stems (r = 0.42), crowns (r = 0.61), and roots (r = 0.66). Allowing 6 wk of regrowth before frost minimized plant density losses for flaccidgrass.

Processed Panax ginseng, Sun Ginseng, Decreases Oxidative Damage Induced by tert-butyl Hydroperoxide via Regulation of Antioxidant Enzyme and Anti-apoptotic Molecules in HepG2 Cells.

Potential antioxidant effect of processed ginseng (sun ginseng, SG) on oxidative stress generated by tert-butyl hydroperoxide (t-BHP) was investigated in HepG2 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase (LDH) leakage test demonstrated that SG dose-dependently prevents a loss of cell viability against t-BHP-induced oxidative stress. Also, SG treatment dose-dependently relieved the increment of activities of hepatic enzymes, such as aspartate aminotrasferase and alanine aminotransferase, and lipid peroxidation mediated by t-BHP treatment in HepG2 cells. SG increased the gene expression of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. However, high dose of SG treatment caused decrease in mRNA level of glutathione peroxidase as compared to low dosage of SG-treated cells. The gene expression of glutathione reductase was found to be slightly increased by SG treatment. In addition, SG extract attributed its hepaprotective effect by inducing the mRNA level of bcl-2 and bcl-xL but reducing that of bax. But, the gene expression of bad showed no significant change in SG-treated HepG2 cells. These findings suggest that SG has hepatoprotective effect by showing reduction of LDH release, activities of hepatic enzymes and lipid peroxidation and regulating the gene expression of antioxidant enzymes and apoptosis-related molecules against oxdative stress caused by t-BHP in HepG2 cells.

Nitrogen loss mitigation using duration-controlled grazing: Field observations compared to modelled outputs

Dairy farmers in New Zealand are encouraged to adopt a range of management strategies, both well established and emerging, to reduce nitrogen (N) losses to waterways. In most regions the OVERSEER® nutrient budgeting software (Version 6) (hereafter referred to as Overseer) is the tool of choice in the assessment of N losses for both regulatory and monitoring purposes. As part of these processes, Overseer is used to assess the impact of improved farm practices on N leaching and runoff from individual farms. In a 3-year dairy system field trial at Massey University, N losses in leaching and runoff under duration-controlled grazing (DC; 4 hours per grazing) were compared with those under standard grazing (SG; 7 hours per day-grazing, 13 hours per night-grazing). A 36% reduction in total nitrogen (TN) losses under DC grazing was measured (14 kg TN/ha) relative to standard grazing (22 kg TN/ha). Farmers adopting DC grazing as a mitigation strategy will only be able to claim the reduction in TN losses estimated by Overseer, and thus observations from the field trial were compared with outputs from Overseer. There was good agreement between the Overseer predictions of N leaching and values measured at the trial site for both the SG and DC grazing treatments. A second Overseer simulation of a DC system suggests that while Overseer is able to predict the reductions in N leaching under DC grazing reasonably well, some issues such as runoff losses and storage of effluent need further consideration. Keywords: Duration-controlled grazing; OVERSEER®; N leaching

6-Thioguanine Reactivates Epigenetically Silenced Genes in Acute Lymphoblastic Leukemia Cells by Facilitating Proteasome-mediated Degradation of DNMT1

Thiopurines including 6-thioguanine ((S)G), 6-mercaptopurine, and azathioprine are effective anticancer agents with remarkable success in clinical practice, especially in effective treatment of acute lymphoblastic leukemia (ALL). (S)G is understood to act as a DNA hypomethylating agent in ALL cells, however, the underlying mechanism leading to global cytosine demethylation remains unclear. Here we report that (S)G treatment results in reactivation of epigenetically silenced genes in T leukemia cells. Bisulfite genomic sequencing revealed that (S)G treatment universally elicited demethylation in the promoters and/or first exons of the genes that were reactivated. (S)G treatment also attenuated the expression of histone lysine-specific demethylase 1 (LSD1), thereby stimulating lysine methylation of the DNA methylase DNMT1 and triggering its degradation via the ubiquitin-proteasomal pathway. Taken together, our findings reveal a previously uncharacterized but vital mechanistic link between (S)G treatment and DNA hypomethylation.

Scorpion in Combination with Gypsum: Novel Antidiabetic Activities in Streptozotocin‐Induced Diabetic Mice by Up‐Regulating Pancreatic PPARγ and PDX‐1 Expressions

The management of diabetes without any side effects remains a challenge in medicine. In this study, antidiabetic activity and the mechanism of action of scorpion combined with gypsum (SG) were investigated. Streptozotocin-induced diabetic mice were orally administrated with scorpion (200 mg kg−1 per day) in combination with gypsum (200 mg kg−1 per day) for 5 weeks. SG treatment resulted in decreased body weight, blood glucose and lipid levels, and increased serum and pancreatic insulin levels in diabetic mice. Furthermore, SG significantly increased the number and volume of beta cells in the Islets of Langerhans and promoted peroxisome proliferator-activated receptor gamma and pancreatic duodenal homeobox 1 expressions in pancreatic tissues. However, scorpion or gypsum alone had no significant effect in this animal model. Metformin showed a slight or moderate effect in this diabetic model, but this effect was weak compared with that of SG. Taken together, SG showed a new antidiabetic effect in streptozotocin-induced diabetic mice. This effect may possibly be involved in enhancing beta-cell regeneration and promoting insulin secretion by targeting PPARγ and PDX-1. Moreover, this new effect of SG offers a promising step toward the treatment of diabetic patients with beta-cell failure as a complementary and alternative medicine.

Snow accumulation in thinned lodgepole pine stands, Montana, USA

Alternative silvicultural treatments such as thinning can be used to restore forested watersheds and reduce wildfire hazards, but the hydrologic effects of these treatments are not well defined. We evaluated the effect of two shelterwood-with-reserve silvicultural prescriptions, one leaving residual trees evenly distributed (SE) and the second leaving residual trees in groups (SG), both with ∼50% basal area removal, on snow accumulation in lodgepole pine stands at the Tenderfoot Creek Experimental Forest, west-central Montana. The snow water equivalent (SWE) close to the seasonal peak was measured at >250 locations in the SE and SG treatments and a control in 2003, 2004 and 2005. Reduced interception in SE resulted in significant ( P < 0.0001) 7.2 and 5.6 cm increases in SWE relative to the control in 2003 and 2004, respectively, and a 1.7 cm increase in 2005. Predictive models for the mean peak SWE in the control and the SE treatment were based on an inverse-exponential relationship between interception efficiency and mean storm intensity, and the average percent deviation for the two models was 9.4 and 7.6%, respectively. In the SG treatment, increased solar radiation and wind resulted in sublimation losses that offset gains due to reduced interception in the openings between groups. Consequently, SG always accumulated significantly less snow than SE ( P < 0.0001), and the SG treatment and control means were not significantly different. Differences in snow accumulation between groups and openings and between the north and south sides of groups meant that the SWE in SG was up to three times more variable than in either SE or the control. The contrasting responses in the SE and SG treatments demonstrate that thinning can have substantially different effects on snow accumulation depending on the spatial arrangement of tree removal.

Using Solanum Glaucophyllum as a Source of 1,25-dihydroxyvitamin D to Prevent Hypocalcemia in Dairy Cows

Reducing cation-anion difference of diets (DCAD) fed just before parturition can prevent milk fever. However this dietary regimen does not entirely eliminate hypocalcemia. Milk fever can also be prevented by exogenous administration of the calcium regulating hormone 1,25-dihydroxyvitamin D. Unfortunately 1,25-dihydroxyvitamin D treatment remains expensive and the pre-partal diets used in most trials would be classified today as high in cations. Solanum glaucophyllum (Sg) is a plant that contains high levels of a glycoside form of 1,25-dihydroxyvitamin D. S. Glaucophyllum is widely distributed in the province of Buenos Aires in Argentina and in Brazil and causes the development of a calcinotic disease in cattle called Enteque Seco. In order to become active the glycoside must be cleaved to liberate the 1,25(OH) 2 D 3 . Rumen microbes are very efficient at this process. Could administration of Sg leaves to cows that were already being fed a low DCAD pre-partum diet further improve calcium status at calving? Nine multiparous Jersey cows were fed a low DCAD diet prior to calving. Urine pH of cows was maintained below 7.0 in all cows the week prior to parturition. Five cows were daily given 2 or 3 g Sg leaves in gelatin boluses beginning 6 days (on average) before calving and continuing for the first 14 days of lactation. None of the four cows fed the low DCAD diet only developed milk fever. Their blood calcium concentration was 7.6, 7.0 and 8.0 mg/dl the day of calving and d 1 and 2 after calving respectively. Cows receiving Sg in addition to low DCAD diet had significantly higher blood calcium concentration during the periparturient period with blood calcium concentrations of 7.8, 8.8 and 9.3 mg/dl the day of calving and d 1 and 2 after calving respectively. Defining subclinical hypocalcemia as blood calcium <7.5 mg/dl, control cows suffered an average of 3 days of subclinical hypocalcemia and the Sg treated cows suffered 0.8 d of subclinical hypocalcemia the first 2 wk of lactation. Thus, Sg treatment improved calcium status in animals that were also being fed a low DCAD diet. Unfortunately, all cows receiving Sg suffered 1-2 days of hypocalcemia (1 cow developed milk fever) between 6 and 8 days after Sg treatment was ended. Mean blood calcium on d 22 of lactation of Sg cows was 6.9 mg/dl compared with 9.2 mg/dl in untreated cows. It appears that Sg treatment supplanted the cow's own calcium homeostasis mechanisms so that abrupt withdrawal of treatment left the cows temporarily unable to control blood calcium concentration. Additional experiments were conducted using a phased withdraw approach. Three cows were treated with Sg from 5 days prepartum to 7 postpartum (2 g Sg/d) from day 8-14 postpartum (1 g Sg/d) and day 15-21 postpartum (0.5 g Sg/d). The phased withdrawal approach resulted in normal calcium concentration for up to 2 weeks following cessation of Sg. These data suggest that Sg can be used successfully in the prevention of hypocalcemia and that a phased withdrawal approach to removing the exogenous source of 1,25-(OH) 2 D 3 was successful in avoiding the hypocalcemic side effects associated with abrupt withdrawal. 242

Effects of Strip Versus Continuous Grazing on Diet Parameters and Performance of Steers Grazing Eastern Oregon Native Flood Meadows

Native flood meadows in the northwestern U.S. are typically managed for hay production. However, management for use by grazing livestock is receiving increased interest. Irrigation water is primarily supplied by surface runoff from snowmelt occurring at higher elevations surrounding the meadows. This experiment was conducted to compare strip (SG) and continuous grazing (CG) effects on livestock diet quality and subsequent performance. On May 1, groups of 20 steers (253 kg) were randomly allotted to two replicates of SG and CG treatment pastures (5.6ha each). Animals in SG pastures were confined by electric fence to an area large enough to provide approximately 65% utilization after a 5- to 7-d grazing period. Average strip size was .5 ha, and varied from .23 to 1.15ha. Herbage allowance was between 305 to 1131 kg/AU under CG, and 72 to 198 kg/AU for SG treatments, as measured on d 1 in each strip.Biweekly esophageal extrusa samples and total fecal collections showed that over the grazing season, average CP of extrusa samples from CG tended to be higher than SG (P = .14), at 13.9 and 10.9%, respectively. Crude protein declined signifi-cantly between May and September. Diet IVDDM varied between treatments (P = .07), at 64.6 and 60.7% on CG and SG, respectively. Grasses were more frequent in extrusa samples from SG than CG (P = .06), at 49 and 35%, respectively. Meadow foxtail was the most common grass on the meadows and was more frequent in SG extrusa samples (P = .05). Forage intake was similar (P = .42) for both treatments, at about 2.1% of body weight per day. Steers were weighed biweekly. Steer average daily gain was greater under CG (P = .09) (1.16 kg/day) than SG (.77 kg/day).

A new Method to Isolate Protoplasts from the Cyanobacterium Anabaena cylindrica

Summary The present study was carried out to establish a new isolation method for cyanobacterial protoplasts. Conventional lysozyme treatment is appropriate for the isolation of spheroplasts from cyanobacteria, but not to produce protoplasts. Using Anabaena cylindrica, Mastigocladus laminosus and Oscillatoria princeps as materials, the effects of various enzyme treatments on protoplast isolation were examined. When Anabaena cylindrica was treated with N-acetylmuramidase SG, protoplasts lacking cell wall residues were obtained. The enzyme digested the layer II of the Anabaena cell wall, as with lysozyme. However, N-acetylmuramidase SG was effective to the digestion of layer IV, but not lysozyme. The treatments with lysozyme, achromopeptidase TBL-1, trypsin and ±-amylase were effective for the digestion of Anabaena cell wall respectively, but led to intracellular structure destruction, and moreover made phycocyanin flow out before the protoplasts were isolated. On the other hand, Anabaena protoplasts produced by N-acetylmuramidase SG treatment were intact in their fine structure and functionally active just after the isolation.

The effects of long term feeding ofSolanum glaucophyllum to growing rats on Ca, Mg, P and bone metabolism

The effects of the long term ingestion of Solanum Glaucophyllum leaves (SG) by the rat was investigated in two series of experiments; the animals were fed a normal (for 8 weeks) or a low Ca diet (for 5 weeks). With both diets, the intestinal absorption of Ca was increased and the endogenous fecal Ca excretion was decreased by SG treatment. Added to a normal Ca diet, SG increased the urinary excretion of Ca, Mg and P and reduced the excretion of hydroxyproline and pyrophosphate. At the histological level, SG induced a higher rate of bone tissue synthesis on trabecular and endosteal surfaces. The bone content of hydroxyproline and citrate increased significantly. The total alkaline phosphatase activity of plasma decreased as a function of SG intake due to a decreased in the activity of the intestinal isoenzyme, which was not compensated by the increase in the bone isoenzyme activity. The Mg absorption was decreased by SG inducing lower Mg balances and lower plasma Mg levels. Added to a low Ca diet, SG increased the severity of the secondary hyperparathyroidism induced by the diet. The urinary excretion of hydroxyproline and the plasma alkaline phosphatase activity (both isoenzymes) were significantly increased. The Na and K content of bone decreased as a function of SG intake. 45Ca kinetic experiments revealed that SG increased the rate of Ca resorption and the rate constant of the fast exchangeable Ca pool, in both diets.