Phenolic contents, antioxidant and anticholinesterase potentials of crude extract, subsequent fractions and crude saponins from Polygonum hydropiper L.

BACKGROUND: We investigated Polygonum hydropiper L. (P. hydropiper) for phenolic contents, antioxidant, anticholinesterase activities, in an attempt to rationalize its use in neurological disorders. METHODS: Plant crude extract (Ph.Cr), its subsequent fractions: n-hexane (Ph.Hex), chloroform (Ph.Chf), ethyl acetate (Ph.EtAc), n-Butanol (Ph.Bt), aqueous (Ph.Aq) and saponins (Ph.Sp) were evaluated for 1,1-diphenyl,2-picrylhydrazyl (DPPH), 2,2-azinobis[3-ethylbenzthiazoline]-6-sulfonic acid (ABTS) free radical scavenging potential. Further, acetylcholinesterase (AChE) & butyrylcholinesterase (BChE) inhibitory activities were performed using Ellman's assay. Moreover, total phenolic contents of plant extracts were determined and expressed in mg of gallic acid equivalent per gram of dry sample (mg GAE/g dry weight). RESULTS: Among different fractions, Ph.Cr (90.82), Ph.Chf (178.16), Ph.EtAc (203.44) and Ph.Bt (153.61) exhibited high phenolic contents. All fractions showed concentration dependent DPPH scavenging activity, with Ph.EtAc 71.33% (IC50 15 µg/ml), Ph.Bt 71.40% (IC50 3 µg/ml) and Ph.Sp 71.40% (IC50 35 µg/ml) were most potent. The plant extracts exhibited high ABTS scavenging ability i.e. Ph.Bt (91.03%), Ph.EtAc (90.56%), Ph.Sp (90.84%), Ph.Aq (90.56%) with IC50<0.01 µg/ml. All fractions showed moderate to high AChE inhibitory activity as; Ph.Cr, 86.87% (IC50 330 µg/ml), Ph.Hex, 87.49% (IC50 35 µg/ml), Ph.Chf, 84.76% (IC50 55 µg/ml), Ph.Sp, 87.58% (IC50 108 µg/ml) and Ph.EtAc 79.95% (IC50 310 µg/ml) at 1 mg/ml). Furthermore the BChE inhibitory activity was most prominent in Ph.Hex 90.30% (IC50 40 µg/ml), Ph.Chf 85.94% (IC50 215 µg/ml), Ph.Aq 87.62% (IC50 3 µg/ml) and Ph.EtAc 81.01% (IC50 395 µg/ml) fractions. CONCLUSIONS: In this study, for the first time, we determined phenolic contents, isolated crude saponins, investigated antioxidant and anticholinestrase potential of P. hydropiper extracts. The results indicate that P. hydropiper is enriched with potent bioactive compounds and warrant further investigation by isolation and structural elucidation to find novel and affordable compounds for the treatment of various neurological disorders.

Phytochemical profile of a blend of black chokeberry and lemon juice with cholinesterase inhibitory effect and antioxidant potential.

In this study, black chokeberry concentrate was added (5% w/v) to lemon juice, since previous reports suggested potential health benefits of this blend. The phytochemical composition, antioxidant capacity (scavenging of DPPH, superoxide and hydroxyl radicals, and hypochlorous acid), and inhibitory activity against cholinesterase of the new blend were determined and compared with those of lemon juice and chokeberry in citric acid (5%). The chokeberry concentrate, rich in cyanidin-glycosides, quercetin derivatives, and 3-O-caffeoylquinic acid, and lemon juice, possessing flavones, flavanones, quercetin derivates, and hydroxycinnamic acids, were characterised. The new drink showed a higher antioxidant effect than the chokeberry or lemon controls for all the tested methods, except for hypochlorous acid, in which lemon juice displayed higher activity. Both the lemon juice and chokeberry controls inhibited acetylcholinesterase and butyrylcholinesterase, and this effect was increased in the new mixtures. The results of the different radical scavenging assays indicate that the lemon-black chokeberry (5% w/v) mixture was more antioxidative than the respective controls separately. Moreover, their inhibition of cholinesterase is of interest regarding neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, or senile dementia.

Risk factors for early death due to recurrence after resection of large hepatocellular carcinomas.

BACKGROUND/AIM: Long-term survival after resection of hepatocellular carcinomas larger than 5 cm in diameter is worse than after resection of smaller tumors. The risk factors for early death due to recurrence after resection of large tumors have not been clearly elucidated. METHODOLOGY: Among 377 patients who underwent curative resection of hepatocellular carcinoma between 1992 and 2004, 115 patients with tumors larger than 5 cm in diameter were enrolled. They were divided into two groups, i.e., 35 patients who died of recurrent cancer within 2 years after surgery and 80 patients who survived for more than 2 years. RESULTS: The preoperative serum alpha-fetoprotein level, positive surgical margins, number of tumors, the serum levels of albumin and alpha-fetoprotein at 1 and 3 months after surgery, as well as the prothrombin time, cholinesterase and protein induced by vitamin K antagonism-II levels at 3 months, respectively, were significant determinants of survival by univariate analysis. Multivariate analysis showed that an albumin level <3.5 g/dl at 1 month and a cholinesterase level <100 U/L at 3 months after surgery were associated with an increased risk of early death due to recurrence. CONCLUSIONS: The hepatic functional reserve in the early postoperative period significantly influences early recurrence and early death after resection of large hepatocellular carcinomas. It is important to maintain a good perioperative nutritional status and early postoperative adjuvant therapy is required for patients with large tumors.

Hepaticojejunostomy--analysis of risk factors for postoperative bile leaks and surgical complications.

Anastomoses between the jejunum and the bile duct are an important component of many surgical procedures; however, risk factors for clinically relevant bile leaks have not yet been adequately defined. The objective of this study was to describe the incidence of bile leaks after hepaticojejunostomy and to define predictive factors associated with this risk and with surgical morbidity. Between October 2001 and April 2004, hepaticojejunostomies were performed in 519 patients in a standardized way. Patient- and treatment-related data were documented prospectively. A bile leak was defined as bilirubin concentration in the drains exceeding serum bilirubin with a consecutive change of clinical management or occurrence of a bilioma necessitating drainage. Surgical morbidity occurred in 15% of patients, the incidence of a bile leak was 5.6%. Multivariate analysis confirmed preoperative radiochemotherapy, preoperative low cholinesterase levels, biliary complications after liver transplantation necessitating a hepaticojejunostomy, and simultaneous liver resection as risk factors for bile leakages, whereas biliary complications after liver transplantation necessitating hepaticojejunostomy, simultaneous liver resection, and diabetes mellitus were significantly associated with postoperative surgical morbidity. Our results demonstrate that hepaticojejunostomy is a safe procedure if performed in a standardized fashion. The above found factors may help to better predict the risk for complications after hepaticojejunostomy.

[A comparative study on the sensitivity and specificity of cholinesterase and glutathione s-transferase in Gammarus pulex L].

Studies on the influences of lindane, pirimiphos methyl, permethrin, zinc and dodecyl linear alkybenzene sulfonate (LAS) on the activity and toxicity of cholinesterase (ChE) and glutathione s-transferase (GST) in Gammarus pulex L. showed that only pirimiphos methyl caused a change in ChE activity in Gammarus, with a significant reduction in enzyme activity after 24 h and 48 h exposure. Both lindane and permethrin caused a change in GST activity in Gammarus, with a significant increase in enzyme activity after 48 h exposure. Lindane alos caused a significant increase in GST activity after 24 h exposure. Biomarkers ChE and GST were demonstrated a high degree of specificity and sensitivity in comparison to the lethality assay, but GST activity was less specific than ChE activity.

Fetal brain transplants induce recovery of male sexual behavior in medial preoptic area-lesioned rats.

Male rats received bilateral lesions within the medial preoptic area which completely abolished sexual behavior. Hypothalamic fetal brain transplants gradually restored sexual behavior to prelesion levels by the 6th week after the transplant. Immunocytochemical analyses revealed tyrosine hydroxylase immunoreactivity neurons within the transplanted tissue. These results demonstrate that fetal brain transplants can restore an innate complex behavior in which no spontaneous recovery is observed.

Distribution of 'non-specific' cholinesterase histochemical staining in the dorsal thalamus: a comparative study in rodents.

Histochemical studies in rat dorsal thalamus demonstrate that 'non-specific' cholinesterase (ChE) enzyme activity is characteristic of neurons of the anterior dorsal (AD) and reuniens (Re) nuclei and in a cell group found as part of the central lateral (CL) and lateral dorsal (LD) nuclei. Extra-somatal ChE staining also is seen in the anterior ventral (AV) nucleus. Parallel histochemical studies in other rodents reveal slight ChE activity in neurons of the mouse AD and LD, but not in other thalamic nuclei. The dorsal thalami of hamsters, gerbils and guinea pigs show no detectable cellular staining of ChE, although low levels of extra-somatal ChE appear in AV and the internal medullary lamina. These data indicate that 'non-specific' cholinesterase activity is not found commonly in neurons of the dorsal thalamus and prominent ChE staining may be unique to the laboratory rat.

Feline islands of Calleja complex: I. Cytoarchitectural organization and comparative anatomy.

Cytoarchitectural analyses demonstrated that the islands of Calleja complex (ICC) is highly developed and discretely organized in the cat. The feline complex is clearly divided into morphological units, each containing a granular Callejal island and a population of satellite neurons. These ICC units change progressively in cytoarchitecture from the lateral to the medial edge of the olfactory tubercle. In particular, the islands flatten, sink into the tubercular molecular layer, and increase in cell density, while their satellite neurons increase in number and decrease in size. The lateromedial transformation was judged to take place in five stages, resulting in the successive appearance of lateral, lateral transitional, central, medial transitional, and medial ICC units. The first two unit types display prominently two additional components of the feline ICC-namely, clusters of dwarf cells and small pyramidal-like neurons constituting the densocellular layer cupping the base of lateral Callejal islands. All of the various types of ICC units contact the tubercular molecular layer via their dwarf and/or granule cell components, raising the possibility of direct olfactory input to the entire Callejal complex (apart from the isla magna). Output from the complex is presumed to arise from the satellite neurons, which are distinguished from adjoining cell populations by their close association with Callejal islands, typical chromophilic character, and relatively large size (15-42 micron in soma length). In the tubercular ICC, these neurons are most numerous immediately above Callejal islands in a fiber-rich zone continuous with the supratubercular zone and hence with the ventral pallidum. In the accumbal ICC, satellite neurons are most conspicuous in granule-cell-poor spaces within the isla magna, where many non-granular neurons are uncharacteristically small and chromophobic. The isla magna itself is unusual not only for its large size but for lateral extensions encircling a group of accumbal neurons far caudally. Such extensions are one of several indications that the isla magna is intimately associated with the nucleus accumbens. A comparative anatomical survey of the ICC in rats, cats, and macaque monkeys demonstrated a number of species differences. Of particular interest is the finding that the complex is unambiguously divided into discrete island-satellite cell units only in cats and macaques. In these species, the complex is also distinguished by a predominance of superficial islands and an especially prominent isla magna. ICC units, however, were most conspicuous in cats.

Distribution of [3H]diisopropylfluorophosphate, [3H]soman, [3H]sarin, and their metabolites in mouse brain.

The brain area distribution of [3H]diisopropylfluorophosphate, [3H]soman, and [3H]sarin and their metabolites in mice was studied after iv administration of sublethal doses. At the appropriate time after the injection of the radiolabeled organophosphate, the mice were decapitated and their brains were dissected into seven areas. There was a relatively even distribution of the parent compounds and their metabolites in all brain areas except the hypothalamus, which contained concentrations of parent compounds the metabolites that were 2-5 times greater than those in other brain areas. Concentrations of the parent compounds and free metabolites declined steadily throughout the time course, whereas concentrations of the bound metabolites remained relatively constant between 6 and 24 hr. There was no correlation between the disposition of soman, and its metabolites, and cholinesterase inhibition in brain areas, which implicates other central mechanisms in the production of organophosphate effects. However, the higher concentrations of organophosphates and their metabolites in the hypothalamus suggest that this area might be important with respect to the pharmacological effects or the toxicity of these compounds.

Serum cholinesterase inhibitors in the commercial hexane impurities.

Commercial hexane was concentrated by distillation. The distillation residue (0.43 ml/l original solvent) contains material which inhibits human serum cholinesterase (ChE) "in vitro" with a slight effect on acetylcholinesterase. Phosphorus was detected equivalent to 0.33 mumol monophosphorus compound/litre original solvent. The inhibition was progressive with the enzyme-inhibitor preincubation time. A partial reactivation of the inhibited enzyme was obtained by treatment with hydroxylamine and 2-PAM. The results are coherent with a covalent inactivation by more than one inhibitor which acylate (probably phosphorylate) ChE, although it seems likely that a reversible but unstable inhibitor could also be present in the hexane residue. The results are discussed in the context of the known neurotoxic effects of n-hexane and some organophosphorus esters.

Localization and properties of the cholinesterase in crustacean muscle.

Cholinesterase (ChE) activity is present in crustacean muscle extracts. However, since acetylcholine (ACh) is not a neuromuscular transmitter in these animals, the role and exact localization of ChE was unknown. The histochemical localization of the enzyme was studied in whole muscle and in the sarcoplasmic reticulum fraction of the extract, 50-microm frozen sections of glutaraldehyde-fixed crayfish tail flexor muscle were incubated with acetylthiocholine (ATC) as substrate, and examined under the electron microscope. After some modifications in published techniques, dense deposits were found associated with the sarcolemma, sarcolemmal invaginations, and transverse tubules. No deposits were found in 10(-4) M eserine, or if butyrylthiocholine (BTC) was substituted for ATC. The vesicles in the sarcoplasmic reticulum fraction which demonstrate the activity must represent minced bits of these membranes. Using a spectrophotometric method, the kinetics of the crustacean muscle enzyme was compared to the acetylcholinesterase (AChE) on mammalian red blood cells and in the lobster ventral nerve cord. Surprisingly, and contrary to previous reports, the crustacean muscle enzyme did not demonstrate substrate inhibition. While a number of similarities to AChE were found, this lack of substrate inhibition makes questionable an unequivocal similarity with classical AChE.