Esters of the Marine-Derived Triterpene Sipholenol A Reverse P-GP-Mediated Drug Resistance

Our previous studies showed that several sipholane triterpenes, sipholenol A, sipholenone E, sipholenol L and siphonellinol D, have potent reversal effect for multidrug resistance (MDR) in cancer cells that overexpressed P-glycoprotein (P-gp/ABCB1). Through comparison of cytotoxicity towards sensitive and multi-drug resistant cell lines, we identified that the semisynthetic esters sipholenol A-4-O-acetate and sipholenol A-4-O-isonicotinate potently reversed P-gp-mediated MDR but had no effect on MRP1/ABCC1 and BCRP/ABCG2-mediated MDR. The results from [³H]-paclitaxel accumulation and efflux studies suggested that these two triterpenoids were able to increase the intracellular accumulation of paclitaxel by inhibiting its active efflux. In addition, western blot analysis revealed that these two compounds did not alter the expression levels of P-gp when treated up to 72 h. These sipholenol derivatives also stimulated the ATPase activity of P-gp membranes, which suggested that they might be substrates of P-gp. Moreover, in silico molecular docking studies revealed the virtual binding modes of these two compounds into human homology model of P-gp. In conclusion, sipholenol A-4-O-acetate and sipholenol A-4-O-isonicotinate efficiently inhibit the P-gp and may represent potential reversal agents for the treatment of multidrug resistant cancers.     

赤霉素对甘蔗发芽和早期蔗茎生长的潜在调节作用

以甘蔗品种CoSe92423单芽为材料,在盘栽条件下研究赤霉素（GA3）对甘蔗生长特点和生化特性的影响。将甘蔗单芽分别浸入10,20,50和100mg·L^-1赤霉素溶液中24h,以清水为对照,将处理的芽段种植于网室泥盘中。结果表明,低水平GA3处理（10,20,和50mg·L^-1）均可刺激蔗芽生长和甘蔗植株的生长,根数、根和叶片鲜重、蔗茎鲜重、叶面积以及茎长均有所增加。与对照相比（70%）,赤霉素可使蔗芽萌发早,发芽率高,其中以50mg·L^-1GA3处理的发芽率最高,为90%。经赤霉素处理的甘蔗株高和茎重分别增加90%～110%、252%～282%,其中以20mg·L^-1GA3处理最高。此外,赤霉素处理的蔗芽中还原糖和IAA含量也有所增加,且均以20mg·L^-1GA3处理最高。因此,低水平赤霉素均对蔗芽萌发和生长具有刺激作用,但高水平赤霉素则对甘蔗生长有抑制作用。     

Anther culture of recalcitrant indica × Basmati rice hybrids

Fertile, green, di-haploid plants were obtained at high frequencies from several indica × Basmati rice F₁ hybrids and/or F₂ plant populations using an improved anther culture procedure. Anthers from cold-pretreated (10 ^°C for 10 d) panicles of six indica (HKR120, HKR86-3, HKR86-217, PR106, Gobind andCH2 double dwarf) and two Basmati rice (Basmati 370,Taraori Basmati) varieties and 14 heterotic indica ×Basmati F₁/F₂ hybrids were cultured in modified agarose-solidified N6M, Heh5M and RZM media. Best callus induction frequencies (2.6–78%) were obtained in RZM medium containing 4% (w/v) maltose,2,4-D, NAA and kinetin. F₂ plants compared to F₁ hybrids and parental rice varieties, were more responsive to anther culture. Androgenesis frequencies of 31–78% were obtained for indica × Basmati F₂ plants in RZM medium in just 30 d which are comparable to or higher than that reported for japonica rice varieties and hybrids involving japonica rice parent(s). Agarose (1.0% w/v)-solidified MS medium containing 3.0% maltose, kinetin, BAP, and NAA, induced green shoot regeneration in 0–51% of the anther-derived callide pending upon the genotype. High plant regeneration frequencies (67–337 green plants per 1000 anthers)were obtained from anther calli of several F₁hybrids (Gobind × Basmati 370 and HKR120 ×Taraori Basmati) and F₂ plants (Gobind × Basmati370, Gobind × Taraori Basmati, HKR86-3 × TaraoriBasmati). A sample of 498 plants obtained from the above hybrids, were transferred to pots with>90% survival; 8–78% of these plants had >5%spikelet fertility and were diploid. In addition,18% of the haploid plants could be diploidized by submerging in 0.1% colchicine solution for 16–18 h. The improved anther culture procedure reported here, resulted in several fold increase in the recovery of green plants from recalcitrant indica × Basmati rice hybrids compared to previous published procedures. The study may accelerate the introgression of desirable genes from indica into Basmati rice using anther culture as a breeding tool. This revised version was published online in July 2006 with corrections to the Cover Date.     

Glycaemic index, glycaemic load and risk of endometrial cancer: a prospective cohort study

OBJECTIVE: High-glycaemic-load diets may increase endometrial cancer risk by increasing circulating insulin levels and, as a consequence, circulating oestrogen levels. Given the paucity of epidemiological data regarding the relationship between dietary glycaemic index and glycaemic load and endometrial cancer risk, we sought to examine these associations using data from a prospective cohort study. DESIGN, SETTING AND SUBJECTS: We examined the association between dietary glycaemic load and endometrial cancer risk in a cohort of 49,613 Canadian women aged between 40 and 59 years at baseline who completed self-administered food-frequency questionnaires between 1982 and 1985. Linkages to national mortality and cancer databases yielded data on deaths and cancer incidence, with follow-up ending between 1998 and 2000. RESULTS: During a mean of 16.4 years of follow-up, we observed 426 incident cases of endometrial cancer. Hazard ratios for the highest versus the lowest quartile level of overall glycaemic index and glycaemic load were 1.47 (95% confidence interval (CI) = 0.90-2.41; P for trend = 0.14) and 1.36 (95% CI = 1.01-1.84; P for trend = 0.21), respectively. No association was observed between total carbohydrate or total sugar consumption and endometrial cancer risk. Among obese women (body mass index >30 kg m(-2)) the hazard ratio for the highest versus the lowest quartile level of glycaemic load was 1.88 (95% CI = 1.08-3.29; P for trend = 0.54) and there was a 55% increased risk for the highest versus the lowest quartile level of glycaemic load among premenopausal women. There was also evidence to support a positive association between glycaemic load and endometrial cancer risk among postmenopausal women who had used hormone replacement therapy. CONCLUSIONS: Our data suggest that diets with high glycaemic index or high glycaemic load may be associated with endometrial cancer risk overall, and particularly among obese women, premenopausal women and postmenopausal women who use hormone replacement therapy.     

Haemato-biochemical,non-specific immunity,antioxidant capacity and histopathological changes in <Emphasis Type="Italic">Labeo rohita</Emphasis> fingerlings fed rubber protein isolate

A 60-day feeding trial was conducted to evaluate the haemato-biochemical, innate immune response, antioxidant capacity and histopathological changes in Labeo rohita fingerlings fed rubber protein isolates (RPI). One hundred and eighty fingerlings (average weight 4.45 ± 0.01 g) were distributed into five experimental groups in triplicate and fed with isonitrogenous and isocaloric diets. Soybean protein isolate (SPI) served as the reference diet (Control), and the treatment diets were formulated as RPI₂₅, RPI₅₀, RPI₇₅ and RPI₁₀₀ replacing 25, 50, 75 and 100% of SPI protein, respectively. The growth performance indices like final body weight (9.54–10.27 g), net weight gain (5.09–5.84 g), metabolic growth rate (4.54–5.02) and feed efficiency ratio (0.60–0.65) among the various groups were not significantly different (P > 0.05). All the haematological parameters, except red blood cells, showed no significant differences compared with the control group (P > 0.05). The immuno-biochemical parameters like albumin, globulin, total immunoglobulin, respiratory burst and lysozyme activities among the various groups did not differ significantly (P > 0.05). The stress enzyme such as superoxide dismutase (SOD), catalase (CAT) and oxidative stress marker malondialdehyde (MDA) showed no significant difference (P > 0.05). Histopathological examination of the liver revealed no marked changes. In summary, the results showed that RPI was well utilised by the fish and its inclusion did not generate any oxidative-induced stress, thus, RPI may be suggested as a potential replacement for SPI in fish diets without any detrimental effects. Hence, protein isolation offers a unique opportunity for the utilisation of rubber seed meal.     

Isolation and biochemical diagnosis of cell lines of groundnut (Arachis hypogaea L) selected on glyphosate†

Three cell lines of groundnut (Arachis hypogaea L), an important oilseed legume, were selected on glyphosate using in-vitro culture techniques. The cell lines isolated through single as well as stepwise selection procedures showed c 20-fold increase in glyphosate tolerance as compared to the unselected control cell line. Studies on the biochemical mechanism of glyphosate tolerance in these cell lines showed a significant increase in the total extractable activity of the target enzyme, 5-enolpyruvyl shikimate-3-phosphate (EPSP) synthase (EC 2.5.1.19), which was further confirmed with immunological data. The over-expressed EPSP synthase activity was, however, subject to inhibition by glyphosate in vitro. Two other key regulated enzymes of the shikimic acid pathway, 3-deoxy-D -arabino heptulosonate 7-phosphate (DAHP) synthase (EC 4.1.2.15) and chorismate mutase (CM) (EC 5.4.99.5) did not show any change in specific activity in the selected cell lines. The enhanced activity of EPSP synthase in the tolerant cell lines was found to be stably inherited in the absence of selection pressure. © 1999 Society of Chemical Industry     

Metabolic and digestive enzyme activity of Pangasianodon hypophthalmus (Sauvage, 1878) fingerlings in response to alternate feeding of different protein levels in the diet

A feeding trial was conducted for 90 days to assess the metabolic and digestive enzyme activity in response to alternate or mixed feeding of different protein levels in the diet of Pangasianodon hypophthalmus fingerlings. One hundred and forty four fingerlings with an average weight of 5.0–5.5 g were distributed into four experimental groups each with triplicate. Four iso‐caloric diets with varying level of crude protein viz., 35%, 30%, 25% and 20% were made and designated as diet 35P, 30P, 25P and 20P respectively. Four treatments were maintained based on four different feeding schedules such as continuous feeding of diet 35P (35P/35P), alternate feeding of 1 day diet 35P next day diet 30P (35P/30P), alternate feeding of 1 day diet 35P next day diet 25P (35P/25P) and alternate feeding of 1 day diet 35P next day diet 20P (35P/20P). As a result, specific growth rate (SGR) and weight gain percentage were relatively higher in treatment (35P/25P) followed by (35P/35P) and (35P/30P). The metabolic enzymes such as lactate dehydrogenase (LDH), malate dehydrogenase (MDH), glucose‐6‐phosphate dehydrogenase (G6PDH), Glucose‐6‐phosphatase (G6Pase), alkaline phosphatase (ALP), acid phosphatase (ACP), alanine aminotransferase (ALT), aspartate aminotransferase (AST) were expressed better in (35P/35P) group followed by (35P/25P) and (35P/30P) groups. The digestive enzymes, protease and lipase activities were higher in (35P/35P) and (35P/25P) groups. The present study revealed that the better nutrient utilization and growth of 35P/25P group is because of their balanced digestive and metabolic enzyme activities through alternate level of higher and lower protein diet.     

Dietary protein level, microbial phytase, citric acid and their interactions on bone mineralization of Labeo rohita (Hamilton) juveniles

A feeding trial was conducted for 60 days to study the effect of dietary protein, microbial phytase and citric acid on intestinal digesta pH, bone ash and bone mineral contents in Labeo rohita juveniles. Eight experimental diets were prepared in 2 × 2 × 2 factorial arrangement with crude protein levels (25% and 35%), microbial phytase (0 and 500 U kg^?1), and citric acid (0 and 3%). The 25% crude protein level feed was supplemented with phytase (U kg^?1) and citric acid (%) at the level of 0,0 (C₂₅); 500,0 (T₁); 0,3 (T₂); 500,3 (T₃), and 35% crude protein level feed at 0.0 (C₃₅); 500,0 (T₄); 0,3 (T₅) and 500,3(T₆) respectively. One hundred and twenty juveniles of L. rohita (av. wt. 12.61–13.72 g) were distributed randomly in eight treatments, each of with three replicates. Addition of citric acid in the 25% crude protein feed significantly decreased (P<0.001) feed pH with concurrent decrease in intestinal digesta pH (P<0.001) and increased the bone ash content (P<0.05) by 4.6%. An interaction between citric acid and phytase (P<0.05) was also observed for bone ash content. Increasing the dietary protein content from 25% to 35% significantly decreased (P<0.01) bone Zn content by 14.9%, which was more prominent with the addition of citric acid, resulting in significant interaction between protein and citric acid (P<0.05), but the bone Cu content was significantly increased (P<0.01) with increasing dietary protein content. Dietary supplementation of microbial phytase (500 U kg^?1) significantly increased (P<0.05) bone Na, Ca, K, P and Fe contents by 15%, 12.1%, 17.4%, 9.2% and 40.7%, respectively, whereas bone P and Mn content was significantly increased (P<0.05) by addition of citric acid (3%). Addition of phytase to plant‐based diets increased the bioavailability of minerals, thereby increasing bone mineralization. The effect of phytase was increased because of addition of citric acid (3%).     

Microarray-based detection and typing of foot-and-mouth disease virus

Foot-and-mouth disease virus (FMDV) is the most economically important veterinary pathogen because of its highly infectious nature and the devastating effects the virus has on the livestock industry. Rapid diagnostic methods are needed for detection and typing of FMDV serotypes and differentiation from other viruses causing vesicular diseases. We developed a microarray-based test that uses a FMD DNA chip containing 155 oligonucleotide probes, 35-45 base pair (bp) long, virus-common and serotype-specific, designed from the VP3-VP1-2A region of the genome. A set of two forward primers and one reverse primer were also designed to allow amplification of approximately 1100 bp of target sequences from this region. The amplified target was labelled with Alexa-Fluor 546 dye and applied to the FMD DNA chip. A total of 23 different FMDV strains representing all seven serotypes were detected and typed by the FMD DNA chip. Microarray technology offers a unique capability to identify multiple pathogens in a single chip.     

Feeding gelatinized carbohydrate in the diets of magur,Clarias batrachus (Linnaeus, 1758): Effects on growth performance,enzyme activities and expression of muscle regulatory factors

This study was conducted to illustrate the effect of dietary gelatinized starch (GS) on the growth performance, enzyme activities and expression of MyoD and Myf5 in magur, Clarias batrachus fingerlings. Four iso‐nitrogenous (37%) and iso‐lipidic (6%) diets containing 15% (D‐1), 25% (D‐2), 35% (D‐3) or 45% (D‐4) GS were fed to 240 fingerlings (2.5 ± 0.5 g) in triplicates groups for 8 weeks. The maximum weight gain %, specific growth rate, protein efficiency ratio, lower FCR and higher mRNA expression of MyoD or Myf5 were found in the D‐3 group fed with 35% GS. Higher hepatosomatic index, viscerosomatic index, body lipid and lower moisture content were found in the D‐4 group. Aspartate transaminase and alanine transaminase activities were found to be higher in the D‐1 group. Amylase, glucose‐6‐phosphate dehydrogenase and blood glucose were higher in the D‐4 group. Superoxide dismutase, catalase and hexokinase activities remain unaffected by the dietary GS levels. Hence, the overall results indicate that 35% GS can improve growth performance and upregulate myogenic regulatory factors, but 45% GS level will favor lipogenesis and compromise growth. Furthermore, Myf5 gene showed more immediate response than the MyoD to the dietary carbohydrate in magur.