Eel acetylcholinesterase inhibition studies with heteroarylphosphinates

The inhibition of eel acetylcholinesterase by the 4-nitrophenyl esters of 2-furyl(methyl)-, methyl(2-thienyl)-, di-2-furyl-, and di-2-thienylphosphinic acid (I, II, III, and IV, respectively) was investigated at pH 6.90 in 0.067 M phosphate buffer (25.0°C) using stopped-flow instrumentation and automated data processing. Our evaluation of the dissociation constant, K_d, the unimolecular bonding rate constant, k₂, and the bimolecular reaction constant, k_i, are the first reported values for these constants for alkyl/heteroaryl and diheteroaryl esters of phosphinic acids. The largest k_i value (19,330 M^?1 sec^?1) was observed for the reaction of I with the enzyme. The order for the remaining three is II > IV > III. There is no direct relationship between the hydrolysis rates of the esters and their anticholinesterase activities on eel acetylcholinesterase. Likewise, there is no direct relationship between their anticholinesterase activities and the LD₅₀ values in rats.     

Effects of energy level and protein source on nitrogen kinetics in steers fed wheat straw-based diets.

A 4 x 4 Latin square metabolism trial with a 2 x 2 factorial arrangement of treatments was conducted to determine N kinetics in steers. Steers were fed either untreated (UNT-WS) or alkaline hydrogen peroxide-treated wheat straw (AHP-WS) based diets supplemented with soybean meal (SBM) or blood meal (BM). Single doses of (15NH4)2SO4 were infused into ruminal pools to determine N kinetics. Ruminal NH3N concentrations (main effects) were 3.81, 1.65, 3.18, and 2.28 mg/dL in steers when fed diets that contained UNT-WS, AHP-WS, SBM, and BM, respectively. Ruminal N pool size was greater (P < .05) for UNT-WS than for AHP-WS diets and also was greater (P < .10) for SBM than for BM diets. Nitrogen flux rate into the rumen was not affected (P > .10) by diet. However, production rate of N from the ruminal pool was greater (P < .05) for UNT-WS than for AHP-WS diets and greater (P < .10) for SBM than for BM diets. Nitrogen recycled into the rumen was 33% greater (P < .05) for AHP-WS than for UNT-WS diets and 26% greater (P < .05) for BM than for SBM diets. Nitrogen recycling (percentage of N intake) was 33, 56, 36, and 49% for UNT-WS, AHP-WS, SBM, and BM diets, respectively. The blood urea N (BUN) concentrations were 10.23, 4.58, 7.15, and 7.65 mg/dL for UNT-WS, AHP-WS, SBM, and BM diets, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Frequency distribution of Echinococcus granulosus hydatid cysts in sheep populations in the Xinjiang Uygur Autonomous Region, China.

Age-prevalence and age-intensity data of Echinococcus granulosus hydatid cysts in sheep populations were collected in an abattoir in the Xinjiang Uygur Autonomous Region, People's Republic of China. The frequency distribution of the larval cysts per sheep was empirically described by the negative binomial model, with parameter k being 0.5273. A mathematical model for the life cycle of E. granulosus was applied to the collected data and the results show that the infection pressure on sheep was 0.4362 (female) or 0.4119 (male) infections per year, the mean number of cysts increased linearly by 0.8824 (female) or 0.9971 (male) cysts every year and acquired immunity was too low to depress this rate of increase. According to certain definitions of steady states for taeniid populations, it was concluded that at least in some parts of Xinjiang, the life cycle of E. granulosus was and may still be in an endemic steady state. Consequently, the regular dog-dosing program would readily drive the infection from an endemic state towards extinction.     

Effects of protein and energy supplementation of wheat straw-based diets on site of nutrient digestion and nitrogen metabolism of lambs.

A 4 x 4 Latin square metabolism trial with 2 x 2 factorial arrangement of treatments was conducted with lambs to determine effects of energy and CP supplementation of wheat straw-based (WS) diets on apparent N digestion, retention, and flow to the abomasum. Four wether lambs (average weight, 32 kg) fitted with ruminal and abomasal cannulas were fed 70 vs 42% WS (remainder of the diet was concentrate) and 9.5 vs 12.5% CP. Ruminal and total tract DM and OM digestion was 41 and 33% greater (P less than .03) for high-energy than for low-energy diets. Apparent N digestibility was greater (P less than .05) for 12.5% CP than for 9.5% CP diets (69.2 vs 62.0%, respectively) and also greater (P less than .03) for high-energy than for low-energy diets (67.4 vs 63.7%, respectively). High-energy diets resulted in a 23% greater (P less than .03) N retention (percentage of N intake) than did low-energy diets; 12.5% CP diets resulted in a 9% greater N retention (P less than .10) than did 9.5% CP diets. Nitrogen retention (percentage of N digested) was 15% higher (P less than .03) for high-energy than for low-energy diets. Protein level had no effect (P greater than .10). Nitrogen retention (grams/day) was 5.65, 6.97, 5.28, and 7.43 for low-energy, high-energy, low-protein, and high-protein diets, respectively; there were responses to energy level (P less than .03) and protein level (P less than .05). Total N flow (grams/day) to the abomasum did not differ (P greater than .10) due to treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Extracts from Black Carrot Tissue Culture as Potent Anticancer Agents

Black carrots contain anthocyanins possessing enhanced physiological activities. Explants of young black carrot shoots were cultured in Murashige and Skoog (MS) medium for callus initiation and were transferred to new MS medium supplemented with four different combinations of 2,4-dichlorophenoxyacetic acid and kinetin. Subsequently, the lyophilized calli and black carrot harvested from fields were subjected to ultrasound extraction with ethanol at a ratio of 1:15 (w:v). Obtained extracts were applied to various human cancer cell lines including MCF-7 SK-BR-3 and MDA-MB-231 (human breast adenocarcinomas), HT-29 (human colon adenocarcinoma), PC-3 (human prostate adenocarcinoma), Neuro 2A (Musmusculus neuroblastoma) cancer cell lines and VERO (African green monkey kidney) normal cell line by MTT assay. The highest cytotoxic activity was achieved against Neuro-2A cell lines exhibiting viability of 38–46 % at 6.25 μg/ml concentration for all calli and natural extracts. However, a significantly high IC₅₀ value of 170.13 μg/ml was attained in normal cell line VERO indicating that its natural counterpart is an ideal candidate for treatment of brain cancer without causing negative effects to normal healthy cells.     

Determination of the role of salicylic acid and Benzothiadiazole on physico-chemical alterations caused by <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> in tomato

Shoe-string disease caused by Cucumber mosaic virus (CMV) is one of the major threats to tomato production worldwide. The alteration in some biochemical parameters in leaves of the susceptible tomato genotype (Nagina) associated with CMV infection and the effect of exogenous application of salicylic acid (SA) and benzothiadiazole (BTH) were studied in this paper. Results showed that exogenous treatment with SA and BTH not only led to plants which gave significantly more yield than diseased controls (DC), but also delayed symptom expression and reduced disease severity. Analysis of biochemical parameters indicated that exogenous application of elicitors and viral infection, significantly affected the activity of peroxidase (POD), superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX). Compared to the DC plants, minimum disease severity and maximum number of fruit were recorded after a single dose of SA + BTH. Maximum plant height was recorded after weekly application of SA and maximum fruit yield per plant was gained with single dose of SA. Moreover, the activity of POD was significantly elicited many-fold after weekly application of SA + BTH, while higher amount of SOD was recorded with single dose of SA. The activity of CAT was also significantly accelerated after weekly application of SA + BTH while increased level of APX was noticed with single dose of BTH. In conclusion, the combined application of SA and BTH played an important role in induction of defense mechanism against CMV infection and can be useful in tomato disease management programs.     

Propagation of Asian isolates of canine distemper virus (CDV) in hamster cell lines

Backgrounds

The aim of this study was to confirm the propagation of various canine distemper viruses (CDV) in hamster cell lines of HmLu and BHK, since only a little is known about the possibility of propagation of CDV in rodent cells irrespective of their epidemiological importance.

Methods

The growth of CDV in hamster cell lines was monitored by titration using Vero.dogSLAMtag (Vero-DST) cells that had been proven to be susceptible to almost all field isolates of CDV, with the preparations of cell-free and cell-associated virus from the cultures infected with recent Asian isolates of CDV (13 strains) and by observing the development of cytopathic effect (CPE) in infected cultures of hamster cell lines.

Results

Eleven of 13 strains grew in HmLu cells, and 12 of 13 strains grew in BHK cells with apparent CPE of cell fusion in the late stage of infection. Two strains and a strain of Asia 1 group could not grow in HmLu cells and BHK cells, respectively.

Conclusion

The present study demonstrates at the first time that hamster cell lines can propagate the majority of Asian field isolates of CDV. The usage of two hamster cell lines suggested to be useful to characterize the field isolates biologically.     

Immunomodulatory effect of recombinant RNA‐dependent RNA polymerase protein of Macrobrachium rosenbergii nodavirus in giant freshwater prawn M. rosenbergii

The present study evaluated the role of recombinant RNA‐dependent RNA polymerase (RdRp) protein of Macrobrachium rosenbergii nodavirus (MrNV) in modulating the immune response and in reducing MrNV load in infected prawn. In the first experiment, prawns (25–30 g) were injected with recombinant RdRp protein (RP) at a concentration of 0, 1.0 and 10 μg, and immune parameters and expression of some immune‐related genes were measured up to 14 days post injection (p.i.). In the second experiment, early juveniles were injected with a similar dose of RdRp and animals were challenged by immersion with MrNV. The infection status was detected in muscles by nested RT‐PCR up to 21 days post challenge. Prawn injected with higher concentration of RP showed significantly higher total haemocyte count at different period post injection. Significant up‐regulation of immune‐related genes was observed within 24 h in prawn treated with lower dose of RP and after 7 days p.i. at higher level of RP injection compared with adult control. Most of the tested samples (63%) were found to be RT‐PCR positive for MrNV at 48 h of post‐immersion challenge. After 14 days, MrNV was detected only in control prawn, while both RP‐injected groups were MrNV negative. This study elucidated the potential viral load reduction role played by RdRP in MrNV‐infected prawn.     

Polyphenolics Profile, Antioxidant and Radical Scavenging Activity of Leaves and Stem of Raphanus sativus L.

Aerial parts (leaves and stem) of Raphanus sativus, which are usually discarded were found to possess potent antioxidant and radical scavenging activity, as measured by standard antioxidant assays. Methanolic and acetone extracts of R. sativus leaves had total polyphenolic content of 86.16 and 78.77 mg/g dry extract, which were comparable to the traditional rich sources such as green tea and black tea. HPLC identification of polyphenolics indicated the presence of catechin, protocatechuic acid, syringic acid, vanillic acid, ferulic acid, sinapic acid, o-coumaric acid, myricetin, and quercetin in leaves and stem. Among the different extraction solvents, methanolic extract of leaves and stem showed potent reductive capacity, significantly inhibited linoleic acid peroxidation and displayed metal chelating activity. Further, they scavenged free radicals effectively with IC₅₀ (half maximal inhibitory concentration) of 31 and 42 μg/ml for DPPH radical, 23 and 52 μg/ml for superoxide radical, 67 and 197 μg/ml for hydrogen peroxide, and 56 and 62 μg/ml for nitric oxide, respectively. Leaves showed most potent antioxidant and radical scavenging activity as compared to stem, which may be accounted for the high polyphenolic content. Leaves and stem of R. sativus, often under-utilized part of this vegetable, thus possessed considerable amount of polyphenolics. Hence, it should be regarded as a potential source of natural antioxidants and could be effectively employed as an ingredient in health or in functional food.