5-Hydroxy-3,6,7,8,3',4'-hexamethoxyflavone induces apoptosis through reactive oxygen species production, growth arrest and DNA damage-inducible gene 153 expression, and caspase activation in human leukemia cells

This study examined the growth inhibitory effects of structurally related polymethoxylated flavones in human cancer cells. Here, we report that 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5-OH-HxMF) induces growth inhibition of human cancer cells and induction of apoptosis in HL-60 cells through modulation of mitochondrial functions regulated by reactive oxygen species (ROS). ROS generation occurs in the early stages of 5-OH-HxMF-induced apoptosis, preceding cytochrome c release, caspase activation, and DNA fragmentation. The changes occurred after single breaks in DNA were detected, suggesting that 5-OH-HxMF induced irreparable DNA damage, which in turn triggered the process of apoptosis. Up-regulation of Bax was found in 5-OH-HxMF-treated HL-60 cells. In addition, a caspase-independent pathway indicated by endonuclease G also contributed to apoptosis caused by 5-OH-HxMF. Antioxidants suppress 5-OH-HxMF-induced apoptosis. 5-OH-HxMF markedly enhanced growth arrest DNA damage-inducible gene 153 (GADD153) protein in a time-dependent manner. N-acetylcysteine (NAC) and catalase prevented up-regulation of GADD153 expression caused by 5-OH-HxMF. These findings suggest that 5-OH-HxMF creates an oxidative cellular environment that induces DNA damage and GADD153 gene activation, which in turn helps trigger apoptosis in HL-60 cells. Meanwhile, ROS were proven an important inducer in this apoptotic process. The C-5 hydroxyl on the ring of 5-OH-HxMF was found to be essential for the antiproliferative and apoptosis-inducing activity. Our study identified the novel mechanisms of 5-OH-HxMF-induced apoptosis and indicated that these results have significant applications as potential chemopreventive and chemotherapeutic agents.     

Gallic acid induces apoptosis in 3T3-L1 pre-adipocytes via a Fas- and mitochondrial-mediated pathway

Gallic acid (3,4,5-trihydroxybenzoic acid) is a naturally abundant plant phenolic compound. Our previous studies have shown that some phenolic acids such as gallic acid inhibit cell growth and induce apoptosis in 3T3-L1 pre-adipocytes. However, the molecular mechanism of gallic acid in the induction of cell apoptosis is still unclear. In this study, we investigated the effect of gallic acid on the apoptotic pathway in 3T3-L1 pre-adipocytes. Western blot data revealed that gallic acid stimulated an increase in the protein expression of Fas, FasL, and p53. The ratio of expression levels of pro- and anti-apoptotic Bcl-2 family members was changed by gallic acid treatment. Gallic acid released mitochondrial cytochrome c into the cytosol and subsequently induced the activation of caspase-9 and caspase-3, which were followed by the cleavage of poly(ADP-ribose) polymerase. Pretreatment with a general caspase-9 inhibitor (Z-LEHD-FMK) and caspase-3 inhibitor (Z-DEVD-FMK) prevented gallic acid from inhibiting cell viability in 3T3-L1 pre-adipocytes. The data also indicated that treatment with gallic acid inhibited histone deacetylase activity in 3T3-L1 pre-adipocytes. These results demonstrate that gallic acid induces apoptosis in 3T3-L1 pre-adipocytes through the Fas and mitochondrial pathway. The induction of cell apoptosis by gallic acid may prove to be a pivotal mechanism for decreased pre-adipocyte proliferation.     

Prevalence of Toxoplasma gondii in small mammals from the Ardennes region, France

Serum samples from 218 small mammals trapped in forest and grassland in the Ardennes region (North-eastern France) were tested for antibodies to Toxoplasma gondii. Using the modified agglutination test, positive results were found in 4/92 Apodemus sp., 3/64 Clethrionomys glareolus, 0/26 Microtus agrestis, 0/4 Micromys minutus, 3/5 Sorex sp., 2/9 Arvicola terrestris, and 7/18 Talpa europaea. Toxoplasma gondii was not isolated from the heart of seropositive individuals after bioassay in mice. Seroprevalence was significantly higher in large fossorial mammals living in grassland than in small forest mammals, probably related to ecological factors.     

Challenges in Adaptation of Plant Disease Warning Systems to New Locations: Re-Appraisal of Billing's Integrated System for Predicting Fire Blight in a Warm Dry Environment

ABSTRACT The purpose of this letter is to describe approaches and possible pitfalls when a fire blight model developed in one climatic area is evaluated in a new location where weather conditions are markedly different. A case is described where a modified form of Billing's integrated system, BIS95, which was developed in a cool moist climate, was tested in a country where weather is warmer and drier. Prior to this, some features of fire blight epidemiology, management, and risk assessment are outlined.     

Clinical and pathologic features of acute bovine liver disease in Australia

Acute bovine liver disease (ABLD) is a sporadic hepatic disease affecting cattle in southern Australia, characterized histologically by striking periportal hepatocellular necrosis. The cause of ABLD is unknown; however, the seasonality and acute presentation of outbreaks suggest mycotoxin involvement. We describe here the clinical and pathologic findings of ABLD in 45 naturally affected cattle from 13 outbreaks occurring from 2010 to 2019 in Victoria, Australia. Outbreaks occurred in herds located along the southern coastal plain of Victoria and were observed most frequently in lactating dairy cattle. Clinical signs commonly included a combination of mild photosensitization, progressive neurologic signs, and hypogalactia, which preceded death by ≤ 48 h. All affected animals had marked elevations in activities of glutamate dehydrogenase, aspartate aminotransferase, and gamma-glutamyl transferase. At autopsy, the most common lesions were serosal petechiae and/or gastrointestinal hemorrhage, and hepatomegaly with a pronounced hepatic reticular pattern. The principal histologic lesion was widespread—severe periportal hepatocellular coagulative necrosis and erythrocyte pooling—which often extended to massive necrosis. Lesions in other organs were uncommon. Our study of ABLD suggests involvement of a potent hepatotoxin that is either directly cytopathic or requires bioactivation by periportal-specific enzymes.     

Treatment of hatchery wastewater by a sequencing batch reactor process

After settling pretreatment, the primary sludge wastewaters from a fish hatchery were treated in three five-litre bench-scale sequencing batch reactors. The sequencing batch reactors were operated concurrently under the same operating conditions. A cycle of 24 hours was adopted during the course of this study. Two different temperature regimes (22°C and 10°C) were employed, and a range of initial BOD₅ concentrations was treated. The results showed the occurrence of very high reductions in BOD₅ and COD concentrations, and very high removal of ammonia---nitrogen and suspended solids. The average five-day BOD₅ reduction was 98% for high influent concentrations, while the reduction for lower influent concentrations was 95%. The average five-day BOD₅ concentration for the effluents was less than 100 mg litre⁻¹. Nitrification was pronounced after 40 days of reactor operation. Denitrification occurred when the reactors were fed wastewaters with high BOD₅ concentrations, although it was not observed with low influent BOD₅ concentrations or at low temperatures (10°C). The overall treatment efficiency remained very high when the reactors were operated under a low temperature regime (10°C).     

Characterization of molecular variants of GnRH, induction of spermiation and sex reversal using salmon GnRH-A and domperidone in the protogynous diandric fish, Synbranchus marmoratus Bloch, (Teleostei, Synbranchidae)

This paper studies the molecular variants of gonadotropin-releasing hormone (GnRH) present in the brain of the protogynous swamp eel, Synbranchus marmoratus, and the effects of the administration of salmon GnRH analogue (sGnRH-A) and the dopamine receptor antagonist, domperidone (DOM) on final maturation and gamete release in this species. Evidence for the presence of two GnRH variants, sGnRH and cIIGnRH were obtained by reverse phase high-pressure liquid chromatography (RP-HPLC) and radioimmunoassay with different antisera. The effects of treatment with sGnRH-A+DOM were checked by three ways: oocyte and milt release by stripping, histological analysis of the gonadal tissue, and androgen serum levels at different times throughout the experiment. In males, spermiation was induced after three weeks of treatment. In the female group, sGnRH-A+DOM did not induce ovulation at the end of the experiment. Histological analysis of the gonads from the female group showed evidence of sex reversal. All the treated fish had elevated androgen serum levels from the third week, with respect to control fish. In all cases, serum estradiol levels were undetectable. These results suggest that treatment with sGnRH analog and DOM induce sex reversal in female and spermiation in males. This revised version was published online in July 2006 with corrections to the Cover Date.     

Integration of independent component analysis with near infrared spectroscopy for evaluation of rice freshness

The storage time and conditions of rice has an enormous effect on its appearance, flavor, and quality of the nutrients; and the acidity of rice usually increases with prolonged storage. Therefore, evaluation of freshness is an important issue for rice quality. In this study, the NIR (near infrared) spectra combined with independent component analysis (ICA) technique was used to evaluate the rice freshness. A total of 180 white rice samples were collected from 6 crop seasons for the purpose of developing an ICA-NIR based procedure for rice freshness as quantified by pH values. Values of pH were determined by a BTB-MR (bromothymol blue – methyl red) method. The best calibration model of white rice was developed using the smoothed first derivative spectra, five ICs and cross-validation; the results indicated that r² (coefficient of determination) = 0.924, and in units of pH, SEC (standard error of calibration) = 0.145, SEP (standard error of prediction) = 0.146, bias = 0.001, and RPD (residual predictive deviation) = 3.65. Freshness of white rice could be distinguished either visually by a 3-dimensional diagram composed from ICs 2, 3 and 4, or statistically by a calibration model. The results show that ICA with NIR has the potential to be adopted as an effective method for evaluating rice freshness.     

Pharmacokinetic studies of ivermectin: effects of formulation

Studies are reported which describe the effects of formulation, animal species, and route of administration on the pharmacokinetics of ivermectin. Biological half-life t1/2 increases in the order: swine (0.5 day) less than dogs (1.8 day) less than cattle approximately equal to sheep (2.8 day). Formulation modifications, based upon the solubility properties of the drug, have been directed towards the development of a nonaqueous injectable formulation for cattle and an aqueous vehicle for horses. Bioavailability following subcutaneous injection in cattle can be regulated by control of injection solvent composition: a vehicle composed of a mixed aqueous-organic solvent exhibits pharmacokinetic properties (i.e., Cp, t1/2, AUC, and F) intermediate between those furnished by an aqueous formulation and via a purely nonaqueous solvent. The longer apparent biological half-life from this latter vehicle (t1/2 = 8.3 days) confirms that a slow absorption process dominates the pharmacokinetics in the nonaqueous injectable product to produce an effective controlled-release formulation. These bioavailability results illustrate the increase in the concentration of an organic solvent and a concomitant decrease in surfactant concentration in a micellar aqueous system for prolonged drug delivery via injection.