In vitro and in vivo Assessment of Cholinesterase Inhibitory Activity of the Bark Extracts of Pterocarpus santalinus L. for the Treatment of Alzheimer’s Disease

Alzheimer’s disease (AD) (a progressive neurodegenerative disorder) is mostly predominant cause of dementia in the elderly. Prolonging the function of acetylcholine by inhibiting both acetylcholinesterase and butyrylcholinesterase is most effective treatment therapy of AD. Traditionally Pterocarpus santalinus L. is widely known for its medicinal use. In this study, in vitro acetylcholinesterase inhibitory activity was investigated and methanolic extract of the plant showed significant activity. To confirm this activity (in vivo), learning and memory enhancing effects were tested in mice. For the test, memory impairment was induced by scopolamine (cholinergic muscarinic receptor antagonist). Anti-amnesic effect of the extract was investigated by the passive avoidance task in mice. The study also includes brain acetylcholinesterase activity. Results proved that scopolamine induced cognitive dysfunction was significantly decreased by administration of the extract solution, in the passive avoidance task and inhibited brain acetylcholinesterase activity. These results suggest that bark extract of Pterocarpus santalinus can be better option for further studies on AD via their acetylcholinesterase inhibitory actions.

Pyrethroid Resistance and Its Mechanism in Field Populations of the Sand Termite, Psammotermes hypostoma Desneux

Termites are eusocial insects that are found on all continents except Antarctica. Termites have serious destructive impact, damaging local huts and crops of poor subsistence. The annual cost of termite damage and its control is determined in the billions globally. In Egypt, most of these damages are due to the subterranean termite species especially the sand termite, P. hypostoma. Pyrethroids became the primary weapon for subterranean termite control, after the use of chlorpyrifos as a soil termiticide was banned. Despite the important role of pyrethroids in termite control, its extensive use in pest control led to the eventual rise of insecticide resistance which may make many of the pyrethroids ineffective. The ability to diagnose the precise mechanism of pyrethroid resistance in any insect species would be the key component of its management at specified location for a specific population. In the present study, detailed toxicological and biochemical studies was conducted on the mechanism of pyrethroid resistance in P. hypostoma. The susceptibility of field populations of P. hypostoma against deltamethrin, α-cypermethrin and ƛ-cyhalothrin was evaluated. The obtained results revealed that the workers of P. hypostoma have developed high resistance level against the tested pyrethroids. Studies carried out through estimation of detoxification enzyme activity indicated that enhanced esterase and cytochrome P450 activities were probably important mechanisms for pyrethroid resistance in field populations. Elevated esterase activity and also additional esterase isozyme were observed in the pyrethroid-resistant populations compared to the susceptible populations. Strong positive correlation between cytochrome P450 activity and pyrethroid resistance was also reported. |Deltamethrin could be recommended as a resistance-breaking pyrethroid that is active against resistant populations of P. hypostoma.

Susceptibility of Spodoptera littoralis, Field Populations in Egypt to Chlorantraniliprole and the Role of Detoxification Enzymes

The cotton leafworm, Spodoptera littoralis (Boisduval) is a major insect pest of vegetables and cotton crops in Egypt, and exhibits different levels of tolerance to certain insecticides. Chlorantraniliprole has been registered recently in Egypt for control this insect. The susceptibilities of three S. littoralis populations collected from El Behaira governorate, north Egypt to chlorantraniliprole were determined by leaf-dipping technique on 4th instar larvae. Obvious variation of toxicity was observed among the laboratory susceptible, and three field populations with LC50 values ranged between 1.53 µg/ml and 6.22 µg/ml. However, all the three field populations were less susceptible to chlorantraniliprole than a laboratory susceptible population. The most tolerant populations were sampled from El Delengat (ED) Province where S. littoralis had been frequently challenged by insecticides. Certain enzyme activity assays were carried out to be correlated with the mechanism of the observed field population tolerance. All field populations showed significantly enhanced activities of detoxification enzymes compared with the susceptible strain. The regression analysis between chlorantraniliprole toxicities and enzyme activities revealed that the highest correlation is between α-esterase or β-esterase (α-β-EST) activity and collected field strains susceptibility, otherwise this correlation is not significant (P > 0.05). Synergism assays showed the ED and susceptible strains could be synergized by known detoxification inhibitors such as piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl-maleate (DEM) at different levels (1.01-8.76-fold and 1.09-2.94 fold, respectively), TPP showed the maximum synergism in both strains. The results show that there is a correlation between the enzyme activity and tolerance, and carboxylic-esterase (Car-EST) is likely the main detoxification mechanism responsible for tolerance of S. littoralis to chlorantraniliprole.

Acute Myocardial Infarction Associated with Ingestion of Herbal Mixtures Containing Acetylcholinesterase Inhibitors: A Case Study

We reviewed an unusual case of a 65-year-old male taking an herbal mixture containing compounds with anticholinesterase activity for a long period of time, presented with acute my myocardial infarction and multiple organ dysfunction syndrome followed by death. Clinically, there are findings correlated with anticholinesterase activity, such as bilateral miosis, diaphoresis, vomiting and fasciculation without a history of any toxic ingestion or exposure. Gas chromatography–mass spectrometry screening studies identified the presence of thymol, anethole in the herbal extract and butylated hydroxytoluene in the blood sample. Hence, with this case report, we intend to highlight the necessity of evaluating the long-term use of the herbal mixture.

The Investigation of Enzymatic Activity in the Soils under the Impact of Metallurgical Industrial Activity in Lori Marz, Armenia

Beta-glucosidase, chitinase, leucine-aminopeptidase, acid phosphomonoesterase and acetate-esterase enzyme activities in the soils under the impact of metallurgical industrial activity in Lori marz (district) were investigated. The results of the study showed that the activities of the investigated enzymes in the soils decreased with increasing distance from the Shamlugh copper mine, the Chochkan tailings storage facility and the ore transportation road. Statistical analysis revealed that the activities of the enzymes were positively correlated (significant) to each other according to the observation sites which indicated that enzyme activities were affected by the same anthropogenic factor. The investigations showed that the soils were polluted with heavy metals (Cu, Pb, As, Co, Ni, Zn) due to copper mining activity in this territory. The results of Pearson correlation analysis revealed a significant negative correlation between heavy metal pollution degree (Nemerow integrated pollution index) and soil enzyme activity. All of this indicated that copper mining activity in this territory causing the heavy metal pollution of the soils resulted in the inhabitation of the activities of the enzymes which are considered as biological catalysts to decompose organic materials and facilitate the cycling of nutrients.

Antioxidative, Anticholinesterase and Anti-Neuroinflammatory Properties of Malaysian Brown and Green Seaweeds

Diminished antioxidant defense or increased production of reactive oxygen species in the biological system can result in oxidative stress which may lead to various neurodegenerative diseases including Alzheimer’s disease (AD). Microglial activation also contributes to the progression of AD by producing several proinflammatory cytokines, nitric oxide (NO) and prostaglandin E2 (PGE2). Oxidative stress and inflammation have been reported to be possible pathophysiological mechanisms underlying AD. In addition, the cholinergic hypothesis postulates that memory impairment in patient with AD is also associated with the deficit of cholinergic function in the brain. Although a number of drugs have been approved for the treatment of AD, most of these synthetic drugs have diverse side effects and yield relatively modest benefits. Marine algae have great potential in pharmaceutical and biomedical applications as they are valuable sources of bioactive properties such as anticoagulation, antimicrobial, antioxidative, anticancer and anti-inflammatory. Hence, this study aimed to provide an overview of the properties of Malaysian seaweeds (Padina australis, Sargassum polycystum and Caulerpa racemosa) in inhibiting oxidative stress, neuroinflammation and cholinesterase enzymes. These seaweeds significantly exhibited potent DPPH and moderate superoxide anion radical scavenging ability (P

Biodegradation of Malathion by Acinetobacter baumannii Strain AFA Isolated from Domestic Sewage in Egypt

Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosa (PS1), and Pseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazinon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 10^12CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Protective Effect of Melissa officinalis L. against Malathion Toxicity and Reproductive Impairment in Male Rats

Malathion (ML) is a well known pesticide commonly used in many agricultural and non-agricultural processes. Its toxicity has been attributed primarily to the accumulation of acetylcholine (Ach) at nerve junctions, due to the inhibition of acetylcholinesterase (AChE). The aim of the current research was to study the protective effect of the melissa plant extract against reproductive impairment induced by malathion in 32 male albino rats, and the biological experiment was divided into four groups (8 in each) that given malathion (27 mg/kg; 1/50 of the LD50 for an oral dose) and/or Melissa officinalis (MO) extract (200mg/kg/day) by gavages technique. The sperm counts, sperm motility, sperm morphology, FSH, LH, and testosterone levels had been determined in testes homogenate at the end of the experiment. It is worthy to report that, rats treated with melissa extract did not show a significant difference when compared with the control group, while rats given malathion alone had significantly lower sperm count, sperm motility, and significantly higher abnormal sperm numbers, than the untreated control rats as well as having significantly lower serum FSH, LH, and testosterone levels compared with the control group. Administrations of melissa extract restore all mentioned histological parameters towards the control group and the melissa extract had a strong positive protective effect against malathion toxicity. Results the of biological parameters were confirmed by the histological examination of rat testes and indicated that, both control and melissa groups showing normal seminiferous tubules, while malathion group testicular tissues had necrosis, edema in the seminiferous tubules and degeneration of spermatogonial cells lining the seminiferous tubules with incomplete spermatogenesis. The use of melissa against malathion improved the histological picture and showing normal seminiferous tubules with complete spermatogenesis and almost there was no histopathological changes could be noted.

Histochemistry of Intestinal Enzymes of Juvenile Dourado Salminus brasiliensis Fed Bovine Colostrum

Enzyme activity was evaluated in the intestine of juvenile dourado (Salminus brasiliensis) fed with diets containing 0, 10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either 30 or 60 days. The intestinal enzymes acid and alkaline phosphatase (ACP and ALP, respectively), non-specific esterase (NSE), lipase (LIP), dipeptidyl aminopeptidase IV (DAP IV) and leucine aminopeptidase (LAP) were studied using histochemistry in four intestinal segments (S1, S2, S3 and posterior intestine). Weak proteolitic activity was observed in all intestinal segments for DAP IV and LAP. The activity of NSE and LIP was also weak in all intestines, except for the moderate activity of NSE in the S2 of 20% LBC group after 30 days and in the S1 of 0% LBC group after 60 days. The ACP was detected only in the S2 and S3 of the 10% LBC group after 30 days. Moderate and strong staining was observed in the first three intestinal segments for ALP and weak activity in the posterior intestine. The activity of DAP IV, LAP and ALP were also present in the cytoplasm of the enterocytes. In the present results, bovine colostrum feeding did not cause alterations in activity of intestinal enzymes.

In vitro and in vivo Anticholinesterase Activity of the Volatile Oil of the Aerial Parts of Ocimum basilicum L. and O. africanum Lour. Growing in Egypt

In this study, the in vitro anticholinesterase activity of the volatile oils of both O. basilicum and O. africanum was investigated and both samples showed significant activity. The major constituents of the two oils were isolated using several column chromatographies. Linalool, 1,8-cineol and eugenol were isolated from the volatile oil of O. basilicum and camphor was isolated from the volatile oil of O. africanum. The anticholinesterase activities of the isolated compounds were also evaluated where 1,8-cineol showed the highest inhibitory activity followed by camphor. To confirm these activities, learning and memory enhancing effects were tested in mice. Memory impairment was induced by scopolamine, a cholinergic muscarinic receptor antagonist. Anti-amnesic effects of both volatile oils and their terpenoids were investigated by the passive avoidance task in mice. We also examined their effects on brain acetylcholinesterase activity. Results showed that scopolamineinduced cognitive dysfunction was significantly attenuated by administration of the volatile oils and their terpenoids, eugenol and camphor, in the passive avoidance task and inhibited brain acetylcholinesterase activity. These results suggest that O. basilicum and O. africanum volatile oils can be good candidates for further studies on Alzheimer’s disease via their acetylcholinesterase inhibitory actions.

Effects of Ciprofloxacin and Levofloxacin Administration on Some Oxidative Stress Markers in the Rat

Fluoroquinolones are a group of antibiotics widely used because of their broad spectrum activity against both Gram-positive and Gram-negative bacteria. In this study, ciprofloxacin and levofloxacin were administered to rats at therapeutic doses to evaluate their effects on plasma arylesterase activity, as well as, on hepatic advanced oxidized protein products (AOPPs) and malondialdehyde (MDA) levels, as measures of oxidative stress. Ciprofloxacin (80 mg/kg body weight) and levofloxacin (40 mg/kg body weight) were administered to male albino rats for 7 and 14 days. The data obtained demonstrated that plasma arylesterase activity was significantly decreased by both drugs with ciprofloxacin administration inhibiting the activity by 29% and 30% while Levofloxacin treatment resulted in 35% and 30% inhibition, after 7 and 14 days treatment respectively. Hepatic AOPP and MDA levels were both elevated by these antibiotics. This study supplies further evidence that fluoroquinolones at therapeutic doses promote oxidative stress.

Enzymatic Esterification of Carboxylic Acids and Higher Alcohols in Organic Medium

The studying of enzymatic esterification of carboxylic acids and higher alcohols was performed by esterase Saccharomyces cerevisiae in water-organic medium. Investigation of the enzyme specificity to acetic substrates showed the best result with acetic acid in esterification reactions with ethanol whereas within other carboxylic acids the esterification decreased with acids: hexanoic > pentanoic > butyric > decanoic. In relation to higher alcohols C3-C5, esterification increased with alcohols propanol < butanol < amylol. Also it was determined that esterase was more specific to alcohols with branched chain such as isobutyl alcohol and isoamyl alcohol. Data obtained may have important practical implications, for example, for application of yeast esterase in producing various volatile esters as well as in enzymatic transformation of volatile acids and toxic fusel alcohols into volatile esters by providing the production of the high quality alcoholic beverages with redused content of higher alcohols as well as with improved degustational and hygienic properties.

Differentiation between Common Tick Species Using Molecular Biology Techniques in Saudi Arabia

Protein and Esterase electrophoresis were used to genetically identify two Saudi tick species. Engorged females of the camel tick Hyalomma dromedarii (Koch) (Acari: Ixodidae) and the cattle tick Boophilus annulatus (Say) (Acari: Ixodidae) ticks collected from infested camels and cattle in the animals resting house at Hail region in KSA were used. The results showed that there are a variation in both of protein and esterase activity levels and a high polymorphism within and between the genera and species of Hyalomma and Boophilus . In conclusion, the protein and esterase electrophoretic analysis used in the present study could successfully distinguish among tick species, commonly found in Saudi Arabia.

Effects of Allelochemical Gramine on Metabolic Activity and Ultrastructure of Cyanobacterium Microcystis aeruginosa

In this study, inhibition of Microcystis aeruginosa by antialgal alleochemical gramine, was studied by analyzing algal metabolic activity (represented by esterase and total dehydrogenase activities) and cell ultrastructure (showing morphological and ultrastructure alterations using transmission electron microscopy and DNA ladder analysis). After gramine exposure, esterase and total dehydrogenase activities were increased firstly but decreased later. In contrast with the controls, the cells exposed to gramine showed apparent ultrastructure alterations with thylakoids in breakage, phycobilins in decrease, lipid and cyanophycin granules abundant firstly but dissolved afterwards, DNA in fragementation. The occurrence of increase of metabolic activity and specific granules reflected that the resistance of cellular response to gramine was initiated. DNA fragementation associated with the increase of metabolic activity and specific granules hinted that gramine caused M. aeruginosa cells to initiate some morphotype of programmed cell death.

Influence of Degradative Enzymatic Activities on the Shelf Life of Ready-to-Eat Prickly Pear Fruits

Prickly pear fruit (Opuntia ficus indica L. Miller) belongs to the Cactaceae family. This species is very sensitive to low storage temperatures (< 5°C) which cause damages. The fruits can be peeled, suitably packaged and successfully commercialized as a ready-to-eat product. The main limit to the extension of the shelf life is the production of off-flavors due to different factors, the growth of microorganisms and the action of endogenous enzymes. Lipoxygenase (LOX) and Pectinesterase (PE) are involved in fruit degradation. In particular, LOX pathway is directly responsible for lipid oxidation, and the subsequent production of off-flavours, while PE causes the softening of fruit during maturation. They act on the texture and shelf-life of post-harvest, packaged fruits, as a function of the the grown of microorganisms and packaging technologies used. The aim of this work is to compare the effect of different packaging technologies on the shelf life extension of ready-to-eat prickly pear fruits with regards for the enzymes activities.