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Antidepressants, antipsychotics, anti-anxiety drugs, or sleeping pills are most often used to treat autism spectrum disorders. The treatment thus focuses on the manifestations accompanying autism, such as inattention, irritability, or sleep disorders, but does not address the essence of the social deficits of autistic children. In addition, drug treatment is often poorly tolerated. Since a certain association of autism with gut microbiota dysbiosis has been demonstrated, and given the existence of the microbiota-gut-brain axis, it can be assumed that modulation of the gut microbiota could contribute to the treatment of the essence of some Autism Spectrum Disorders category manifestations. Here, we present an example of the use of the nutritional supplement Juvenil to alleviate the manifestations of Autism Spectrum Disorders in a patient with Asperger’s syndrome.

Authors of the paper assess physical activities on selected salutoprotective factors, which have demonstrable relationship towards processes of health/disease (thus burnout syndrome). The research was implemented on students of 1^st and 2^nd year of social work field during the year 2016/2017. The monitored sample consisted of 46 students of social work at the age of 19 up to 22 years. The Wellness Inventory by authors Travis and Ryan (2004) (24) questionnaire was used for the purpose of this work, which depicts subjective assessment, approaches and motivation of the stated individual in terms of physical activities. There were methods used in order to objectify the quality of subjective perception during the period of one term (SUPOS 7), individual disposition to cope with stress and living sense of life (non-tolerance SOC) and strategies of stress coping (SVF 78).  According to values of this test the respondents were divided into two groups (a group of sportsmen and a group of non-sportsmen). The group of sportsmen showed a higher level of invincibility type SOC and meaningfulness, more positive strategies for coping with stress prevailed, a higher value of activating block of subjective perception was seen.

Organophosphorus compounds (e.g. nerve agents - sarin, VX or tabun) cause deleterious intoxications via inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) leading to cholinergic crisis or even death of intoxicated person. Their causal treatment is managed by oxime reactivators (e.g. pralidoxime, obidoxime, asoxime), which are primarily restoring function of AChE via nucleophilic oximates and thus safe a life.
Recently, we have proved that nucleophilicity of oxime reactivators may be improved by using other substituents attached to the close proximity of the oxime moiety. Such modifications are leading to the increased formation of oximate nucleophile and thus to the increased reactivation of phosphylated cholinesterases. In particular, the substitution by chlorine atoms led to the powerful and rapid reactivation of sarin, cyclosarin or VX-inhibited AChE in vitro (1) or in vivo (2). On the other hand, the substitution by fluorine atoms led to even higher nucleophilicity, but also rapid depletion of oximes into isoxazole degradation products (2-3). Interestingly, some further modified reactivators were proved to be effective in vitro to simultaneously restore function of AChE and BChE.

Haemonchus contortus, a widely distributed parasitic nematode of ruminants, has become resistant to most anthelmintic classes. Therefore, there has been a major demand for new compounds effective against H. contortus and related nematodes. Previous phenotypic screening and further testing have revealed one compound, substituted benzamide BLK127, active against H. contortus larvae and adults (1,2). Based on these results, 13 derivatives of this compound were designed and synthetized. The aim of the present study was to assess the toxicity of these derivatives in H. contortus eggs and adults and their potential hepatotoxicity in sheep in vitro. Isolated eggs and adults from sheep experimentally infective with H. contortus of three strains (one drug-sensitive and two drug-resistant) and precise cut ovine liver slices were used. Only one compound significantly inhibited the egg hatching. On the other hand, four compounds significantly decreased the viability of H. contortus adults at micromolar concentrations. Some of them were effective not only in the drug-sensitive strain but also in the drug-resistant strains. None of the derivatives exhibited a hepatotoxic effect, even at the highest concentration tested (100 µM). Based on these findings, the two most potent compounds were selected for further testing in vivo.

Poisoning from chemical warfare agents (CWAs) such as sarin is associated with neuronal degeneration. This damage is thought to result from glutamatergic excitotoxicity such as seen following kainic acid induced seizures. In order to search for novel neuroprotectants it is necessary to select good mouse models for susceptibility to nerve agent-induced seizures and the resulting neurodegeneration. The mouse strains tested (C57BL/6, ICR, DBA/2, SW, and FVB/N, Harlan Laboratory) had widely different sensitivity to sarin as shown by differences in the dose required resulting in 50% mortality, LD₅₀. Differences also were observed among the strains in Fluoro-Jade C staining with the C57BL/6 and DBA having little to no staining when euthanized at 7 days whereas the other strains did. Differences in acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activity were found among the strains as well. The ICR strain was excluded from the FOB and weight data due to difficulty getting a consistent LD₅₀. Weight loss and FOB scores were similar for all strains. All strains had inhibited AChE activity after sarin exposure and exhibited inhibition of CNS BuChE after sarin exposure but only ICR and SW reached significance.

The best-known function of acetylcholinesterase (AChE) is the hydrolysis of the neurotransmitter acetylcholine, however we are increasingly aware of the multifunctionality of this enzyme [1]. The non-hydrolytic functions of AChE are driven by allosteric sites as the peripheral allosteric site (PAS) responsible for amyloidosis in Alzheimer’s disease through interaction with β-amyloid peptide. We would like to show our work about the identification and characterization of new allosteric sites in AChE, using computational tools. This study has allowed us to identify allosteric inhibitors by virtual screening using our in-house MBC chemical library [2] guided by structure-based and fragment hotspot strategies. The identified compounds were also screened for in vitro inhibition of AChE and three of them were observed to be active. Further experimental (kinetic) and computational (molecular dynamics) studies have been performed to verify the allosteric activity. Thus, new compounds have been developed as allosteric modulators that may be valuable pharmacological tools in the study of non-cholinergic functions of AChE.

The mechanism of toxicity from acute exposure to organophosphorus toxicants (OP) is understood.  Thousands of publications have confirmed that AChE inhibition results in muscle weakness and respiratory failure.  AChE activity returns to normal levels within one month, but symptoms can persist for a lifetime.  For example, people exposed to sarin in the 1995 Tokyo subway attack still have adverse symptoms 23 years later.  Farmers and sheep dippers exposed to OP pesticides have an elevated risk of psychiatric disorders and suicidal behavior.  Epidemiology studies show an association between OP exposure and Alzheimer’s disease and Parkinson’s disease.  We propose a mechanism to rationalize these observations independent of cholinesterase inhibition.  Mass spectrometry analysis of OP-treated proteins shows that OP make stable adducts on tyrosine and lysine.  Furthermore, we have mass spectrometry evidence that OP-lysines promote crosslinks between proteins.  The crosslinked proteins are visualized as protein aggregates on SDS gels and Western blots.  Mass spectrometry has identified γ-glutamyl-ε-lysine and aspartyl-ε-lysine isopeptide bonds between crosslinked peptides.  We propose, but have not yet proven, that isopeptide crosslinked proteins form stable, insoluble aggregates in the brain, similar to the protein aggregates found in Alzheimer’s, Parkinson’s, and prion diseases.  In summary, we propose that chronic neurotoxicity from OP exposure is initiated by OP-lysine formation followed by protein aggregation.  Our proposed mechanism could apply to a variety of compounds and lead to an understanding of neurotoxicity induced by many chemicals.

Carbamates are esters of substituted carbamic acids that react with acetylcholinesterase (AChE) in a two-step process, with initial transfer of the carbamoyl acyl group to a serine residue of AChE accompanied by loss of the carbamate leaving group followed by hydrolysis of the carbamoyl enzyme.  This hydrolysis, or decarbamoylation, is relatively slow, and half-lives of carbamoylated AChEs range from 4 min to more than 30 days.  Since carbamates are poor, slowly reversible AChE substrates, they are effective AChE inhibitors that have been developed as insecticides and therapeutic agents.  We show that decarbamoylation rates are independent of the leaving group for a series of carbamates with the same carbamoyl group.  For a given leaving group, when the alkyl substituents on the carbamoyl group increased in size from N-monomethyl- to N,N-dimethyl-, N-ethyl-N-methyl-, or N,N-diethyl-, the decarbamoylation rates decreased by 4-, 70-, and 1000-fold, respectively. Thus the larger the size of the alkyl groups, the slower the rate of decarbamoylation due to active site distortion.  Furthermore, solvent deuterium oxide isotope effects for decarbamoylation decreased from 2.8 for N-monomethylcarbamoyl AChE to 1.3 for N,N-diethylcarbamoyl AChE, indicating a shift in the rate-limiting step from general acid-base catalysis to a likely conformational change.

Despite extensive oxime research in the last 60 years pralidoxime is still the standard oxime in e.g. United States, British and French forces and obidoxime standard therapy for OP poisoning in several European countries. Oxime research focusses on highly potent oximes with activity against selected nerve agents, broad-spectrum oximes with activity against relevant nerve agents and centrally active (non-)oximes but virtually no compound brought significant improvements compared to the established obidoxime and pralidoxime. In the US MMB-4 is sought to replace pralidoxime and in Germany, France, UK, Canada and other European countries HI-6 is in advanced development for use as nerve agent antidote. Yet, both compounds are not considered as broad-spectrum antidotes and as a mid-term solution combinations of oximes in service with overlapping reactivation potency e.g. obidoxime and HI-6 have been proposed. We here set out to analyze the combination of obidoxime and HI-6 in both a static and dynamic model against poisoning with nerve agents and organophosphorus compound pesticides in vitro. In a cuvette based system the combination of HI-6 and obidoxime both 30 μM for sarin-, cyclosarin-, tabun-, VX- and paraoxon-inhibited human AChE did not result in an impaired reactivation compared to the sole use of both oximes but in a broadened spectrum. Similar results were gained with a dynamic model allowing simulation of nerve agent and pesticide toxicokinetics and oxime pharmacokinetics resembling in vivo conditions. Additional experiments in species closely related to humans e.g. swine are necessary to analyse a potential benefit in vivo.

Organophosphorus (OP) compounds are potent acetylcholinesterase (AChE) inhibitors that have found use as both chemical warfare agents (CWAs) and as pesticides. Following inhibition of AChE by OP compounds, a competitive dealkylation reaction of the phosphylated serine residue occurs – a process referred to as aging. Current therapeutic reactivators of OP-inhibited AChE, mainly oximes, are not effective once aging has occurred. For the first time, we have demonstrated in vitro conversion of the aged AChE to the native form using small drug-like molecular therapeutics.  As part of this effort, a diverse library of small molecule therapeutics have been developed to both recover the activity of aged-AChE, termed resurrection, as well as the activity of inhibited-AChE, referred to as reactivation. The structure of such therapeutics is derived from pyridyl-based quinone methide precursors (QMPs), sharing structural similarities to known therapeutic oximes. A structure-activity relationship study of synthesized QMP therapeutics was conducted to determine the effect electron-donating and electron-withdrawing groups have on the efficiency of both processes and to design optimized small molecule therapeutics for in vivo biological efficacy. Our successes will be presented.

Trichloronate is a racemic organophosphatioate insecticide. It induced delayed neuropathic in hens and human. The avian are species with greater susceptibility to organophosphorus poisoning due to their low levels of A-esterases. However, a copper-dependent hydrolyzing activity of hexyl dichlorophenyl phosphoramidate (HDCP), known as “antogonistic stereoselectivity” was recently identified in chicken serum. This study shows the activating effect of copper on the hydrolysis of trichloronate enantiomers by turkey serum and albumin (TSA) using chiral chromatography with CHIRALCEL OD column and heptane HPLC as mobile phase. The trichloronate hydrolysis levels (µM remaining concentration of each isomer) quantified at 37 °C, pH 7.4 and 60 minutes of turkey serum (10 μL) incubated with 300 μM of copper were statistically higher p˂0.05) for (-)-trichloronate (65 %) than (+)-trichloronate (32%). This estereoselective hydrolysis observed in turkey serum was confirmed by the incubation of 200 μg of turkey serum albumin (amount of this protein estimated in the 10 mL of turkey serum) with 400 μM of racemic trichloronate and 300 μM of copper at physiological condition during 60 minutes; hydrolysis values of 90% and 72% were obtained for (-)-trichloronate and (+)-tricholoronate. In conclusion, the present study evidences the hydrolysis of an organophosphatioate racemic for an A-esterase activity in turkey serum and identifies albumin as the cuproprotein responsible of this Cu²⁺-dependent stereoselective hydrolysis of this chiral insecticide in the turkey serum.

Small ionizable, zwitterionic oximes of limited toxicity show successful outcomes in non-human primates upon intramuscular post-treatment of exposures to OP’s, that enter via the respiratory tract.  Along with their inherent limitations, we consider the bases for success in post-exposure treatment of OP toxicity and reversal of OP-induced sequelae of symptoms¹.  (1) High vapor pressure OPs carry the largest acute exposure risk in mass terrorism.  Toxic OPs released from explosive devices or into controlled ventilation environments are governed by partial pressure and Fick’s Second Law of Diffusion (inverse square of the distance);  (2) Low molecular weight, zwitterionic oximes confer optimal nucleophile orientation and activity within the confines of the OP-impacted, active center gorge of human acetylcholinesterase (AChE).  (3) We emphasize features of ionizable neutral oximes of low toxicity that allow facile passage of membranes to peripheral and central AChE targets and optimal attack angles in the AChE active center.  Hence, for volatile OP’s, antidotes must rapidly enter the circulation, post-exposure, to chase the offending OP.  Following entry, antidotes should then hastily equilibrate between tissue compartments and cross the blood-brain barrier.  Accordingly, we examine the ionization states of zwitterionic oximes and other cationic and anionic (F^-) nucleophiles in relation to their kinetic parameters of reactivation².  Toxicities, both realized and potential, of nucleophilic antidotes in different ionization states, and pharmacokinetics in mice and macaques, under control and exposure conditions, emerge as critical factors for determining in vivo antidote efficacy.  Data will be presented on multiple OP’s and their enzyme conjugates^1-3, comparator oximes and in three animal species/strains.

Chiral analogous compound of methamidophos insecticide are only poorly hydrolyzed by Ca²⁺-dependent phosphotriesterases in mammals tissues including the human serum. We reported the hydrolysis of O-hexyl O-2,5-dichlorophenyl phosphoramidate (HDCP) in chicken serum. The hydrolysis of the R-(+)-HDCP isomer is strongly increased in vitro in the presence of 30-250 µM copper.  It is the opposite estereoselectivity of that showed by liver Ca²⁺-dependent activity. We name it as "antagonistic stereoselectivity". Diluted chicken serum (10 µL in 1 mL solution of 400 µM HDCP) or the equivalent amount of commercial chicken serum albumin (CSA 216 µg/mL) with 100 µM Cu²⁺, showed about 50% and 75% of R-(+)-HDCP hydrolysis after 60 and 120 min. In the same conditions other commercial serum metalloproteins with high affinity to Cu²⁺ (cuproproteins) as human serum ceruloplasmin or horse kidney metallothionein did not showed significant Cu²⁺-dependent hydrolysis. Moreover, other divalent cations (Zn²⁺, Fe²⁺, Ca²⁺, Mn²⁺ and Mg²⁺) did not showed this activation. The results confirm that the CSA is the protein responsible of "antagonistic stereoselectivity" that had been observed in the chicken serum. The effect of copper on the hydrolysis of HDCP by other animal albumins is shown in this work.

Malaria is transmitted by the Anopheles gambiae mosquito in sub-Saharan Africa and tropical regions where the disease is prevalent.  Indoor spraying with anticholinesterase insecticides is a proven method to control populations of the mosquito and to reduce spread of the disease; however,  widespread use of insecticides has led to the rise of an insecticide-resistant G119S mutant acetylcholinesterase in the mosquito which threatens ongoing disease-control efforts.  We have solved high resolution X-ray structures of the G119S mutant acetylcholinesterase of An. gambiae (G119S AgAChE), in the ligand-free state and in complex with a potent difluoromethyl ketone inhibitor, revealing the structural basis of insecticide resistance².  Although resistance-breaking inhibitors of G119S AgAChE exist, they also inhibit human acetylcholinesterase and thus lack the necessary species selectivity to be safely used as insecticides.  In our structures, we see specific features within the active site gorge, including an open “back door”, that are distinct from human acetylcholinesterase.  These differences provide a means for improving species-selectivity in the rational design of improved insecticides for malaria vector control.

A new family of indazolylketones with a multitarget profile as modulators of cholinergic and BACE-1 enzymes and  cannabinoids receptors [1] was  designed based on our previous results [2]. We present the synthesis, computational studies and biological evaluation and of a new family of heterocyclic compounds. Pharmacological evaluation include in vitro inhibitory assays in AChE/BuChE enzymes and BACE-1. In addition, functional activity for cannabinoid receptors has been carried out. The results of the pharmacological tests have revealed that some of these derivatives behave as CB2 cannabinoid agonists and simultaneously show BuChE and/or BACE-1 inhibition. Furthermore, studies in human neuroblastoma SH-SY5Y cells and in the lymphoblasts of patients with Alzheimer's disease have shown neuroprotective effects of this family of compounds, as well as their capacity to blunt the abnormal enhanced proliferative activity of AD lymphoblasts. Based on the in vitro and functional studies we performed in vivo studies of those best compounds employing transgenic mouse (TgAPP) model. The results of the in vivo study revealed that some of these compounds could be very promising candidates for the treatment of Alzheimer's disease.

Neurodegenerative diseases are multifactorial. Therefore, their treatment requires drugs that can act simultaneously on multiple pathogenic targets. We synthesized several series of hybrid structures combining certain pharmacophores essential for neurodegenerative disease treatment: γ-carbolines, carbazoles, phenothiazines, and aminoadamantanes [1-3]. Inhibitory activity of these conjugates against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and carboxylesterase (CaE) was studied along with their ability to competitively displace propidium iodide from the peripheral anionic site of electric eel AChE to assess their potential effect on AChE-induced aggregation of β-amyloid. Antioxidant properties were examined computationally with density functional theory and measured experimentally using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and oxygen radical absorbance capacity (ORAC-FL) assays. Binding modes of conjugates to AChE and BChE were studied using quantum mechanical-assisted molecular docking. Results revealed structures that were selective inhibitors of BChE [1,2] or that combined high potency and selectivity toward BChE with high radical-scavenging activity, e.g., conjugates of γ-carbolines and tetrahydrocarbazoles [3]. Conjugates of γ-carbolines and cycloalcaneindoles with the phenothiazine derivative Methylene Blue demonstrated high potency against AChE and BChE combined with effective displacement of propidium from the peripheral anionic site of AChE. Additionally, the conjugates were extremely active in both antioxidant tests. All conjugates were poor CaE inhibitors and therefore expected to lack drug-drug interactions by this pathway. Good agreement was found between experimental and computational results. Lead compounds were identified for future optimization and development of new multi-target drugs against neurodegenerative diseases that combined cognition enhancement with neuroprotective potential.

N-methyl-D-aspartate receptors (NMDARs) are glutamate-gated ion channels that mediate excitatory neurotransmission in the mammalian central nervous system (CNS), but their dysregulation results in the aetiology of many human CNS disorders. Several NMDAR modulators including memantine have been used successfully in clinical trials. Indeed, 1,2,3,4-tetrahydro-9-aminoacridine (tacrine; THA) was the first approved drug for Alzheimer’s disease (AD) treatment. 7-methoxyderivative of THA (7-MEOTA) is less toxic and showed promising results in patients with tardive dyskinesia. Here, we employed electrophysiological recordings in HEK293 cells and rat neurones to examine the mechanism of action of THA and 7-MEOTA at the NMDAR. We showed that both THA and 7-MEOTA are “foot-in-the-door” open-channel blockers of GluN1/GluN2 and GluN1/GluN3 NMDARs and that 7-MEOTA is a more potent but slower blocker than THA. Furthermore, the inhibitory potency of 7-MEOTA at synaptic and extrasynaptic hippocampal NMDARs was similar, and 7-MEOTA exhibited better neuroprotective activity in rats exposed NMDA-induced lesions in hippocampus when compared with THA and memantine. Finally, intraperitoneal administration of 7-MEOTA attenuated MK-801-induced hyperlocomotion in rats. We conclude that 7-MEOTA is a promising candidate for the treatment of diseases associated with the dysfunction of NMDARs.

Female Wistar rats were percutaneously (p.c.) intoxicated (1xLD₅₀) with VX and its two derivatives differing in their substitution on nitrogen (diethyl- and dibutyl- derivatives). Blood cholinesterase activity was continuously monitored; 100 min after the intoxication (or after death), acetylcholinesterase (AChE)  activity was determined in diaphragm and brain parts (pontomedullar area - PM, frontal cortex - FC and basal ganglia – BG). Blood ChE activity remains unchanged at very short interval (5 min) after VX administration; this interval was prolonged for diethyl- and dibutyl derivatives. AChE activity was decreased to 20-30% of control values in diaphragm, then in FC (60-70%) and PM (54-74%). AChE activity in BG was relatively resistant (cca 80%). When the AChE activity was compared for all three agents in relationship to survival (11 animals) or death (7 animals), significant differences between the activities in survived (32%) and died (13%) rats were demonstrated in diaphragm but not in the blood. This tendency (higher AChE activity in survived animals) was also observed in PM and FC, however, not statistically significant. It is concluded that substitution on nitrogen atom probably influences penetration through the skin; the rest of agent molecule (phosphorus head) probably influences AChE inhibition. As hypothesis, AChE activity in diaphragm could be important for survival  or death in case of p.c. intoxication with these types of V agents.

The acute toxicity of OPNA results from irreversible inhibition of AChE (EC 3.1.1.7), a key enzyme in neurotransmission, via the formation of a covalent P–O bond at the catalytic serine. Inhibition of AChE leads to the accumulation of acetylcholine neurotransmitter (ACh) in the synaptic cleft causing among other symptoms, seizures and respiratory arrest leading to death.  The current urgency treatment of OPNA poisoning is based on the administration of a cocktail of three components: an antimuscarinic agent (e.g. atropine), an anticonvulsant drug (e.g. diazepam) and mono or bispyridinium AChE reactivator (e.g. pralidoxime, obidoxime, trimedoxime). The high nucleophilicity of these alpha-nucleophiles allows the displacement of the phosphyl group from the catalytic serine, yielding to the restoration of AChE activity.  However, reactivation of central AChE is inefficient due to the fact that positively charged pyridiniums poorly cross the brain blood barrier (BBB). Moreover pyridinium(s) oximes exhibit a quite narrow spectrum of reactivation. Despite decades of research in this field, there are no efficient and general broad-spectrum reactivators for OP-inhibited AChE.  In this context, we have developed families of new uncharged reactivators of OP-inhibited acetylcholinesterase and/or OP-inhibited butyrylcholinesterase with the potential to cross the BBB. Three new families of uncharged reactivators display in vitro reactivation potencies towards VX-, tabun- and paraoxon-inhibited human AChE that are superior to those of the mono- and bis-pyridinium aldoximes (e.g. 2-PAM, HI-6, obidoxime, HLö-7, TMB-4) which include those currently used in the armed forces.

Several flavonoids have been identified to induce the expression of AChE in PC12 cells, e.g. daidzin, irisflorentin, cardamonin and genistein. Among them, genistein is the most robust inducer for AChE activity. Genistein, 4',5,7-trihydroxyisoflavone, is a major isoflavone in soybean, which is known as phytoestrogen having known benefit to brain functions. Being a common phytoestrogen, the possible role of genistein in the brain protection needs to be further explored. In PC12 cells, application of genistein significantly induced the expression of neurofilaments, markers for neuronal differentiation. In parallel, the expression of tetrameric form of proline-rich membrane anchor (PRiMA)-linked acetyl-cholinesterase (G4 AChE), a key enzyme to hydrolyze acetylcholine in cholinergic synapses, was induced in a dose-dependent manner: this induction included the associated protein PRiMA. Genistein-induced AChE expression was fully blocked by the pre-treatment of H89 (an inhibitor of protein kinase A) and G15 (a selective G protein-coupled receptor 30 (GPR30) antagonist), which suggested a direct involvement of a membrane-bound estrogen receptor-GPR30-in the cultures. In parallel, the estrogen-induced activation of GPR30 induced AChE expression in a dose-dependent manner. The genistein/estrogen-induced AChE expression was triggered by a cyclic AMP responding element (CRE) located on the ACHE gene promoter. The binding of this CRE site by cAMP response element-binding protein (CREB) induced ACHE gene transcription. We have shown for the first time the activation of GPR30 could be one way for estrogen or flavonoids, possessing estrogenic properties, to enhance cholinergic functions in the brain, which could be a good candidate for possible treatment of neurodegenerative diseases.

Acetylcholinesterase (EC3.1.1.7; AChE) is one of the most important enzymes in the cholinergic system. Our previous works showed that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a notorious persistent organic pollutants, suppressed neuronal AChE activity by both transcriptional and post-transcriptional regulations via aryl hydrocarbon receptor pathway in SK-N-SH human neuroblastoma cells [1, 2]. In the nervous system, the most abundant cell type, astrocyte is regarded to play vital roles in protecting neurons from various kinds of insults, including environmental pollutants. Astrocytes have been considered as one of the target cells of dioxin in the nervous system. However, whether astrocytes are able to mediate indirect effect of dioxin on neuronal AChE is still unknown. In the present study, we aimed to reveal the potential indirect effect by using conditional medium derived from dioxin-treated astrocytes. Rat primary astrocytes were employed which were exposed to TCDD at 0.01 to 1 nM directly for 4 days. After the treatment, the astrocyte conditioned medium (ACM) was collected and administrated to the primary neurons on DIV (day in vitro) 2 for 4 days. Meanwhile, the primary neurons (DIV 2) from the same bench were exposed to TCDD directly at same concentrations for 4 days. The results showed that the enzymatic activity and mRNA expression of AChE was suppressed in TCDD-ACM-treated neurons compared to those of solvent-ACM-treated neurons. The effective concentrations of TCDD were 0.01 and 0.03 nM, which are close to the average serum TCDD concentration in exposed population from different areas of the world. However, AChE was less sensitive in the primary neurons directly exposed to TCDD. These results suggested that astrocytes play roles in mediating the indirect effect of TCDD on neuronal AChE expression.

Acetylcholinesterase (AChE) hydrolyses acetylcholine that functions as a neurotransmitter in neurons. The non-neuronal functions of AChE have been proposed in different cell types. Here, we revealed the expression of AChE in melanocyte and melanoma, in which the tetrameric (G4) form was the major isoform. In the melanogenesis of cultured B16F10 murine melanoma, the amount of AChE was markedly decreased. The differentiation of melanoma led to: (i) increase of melanin and its synthesis enzyme tyrosinase; (ii) change of intracellular cAMP level; and (iii) decrease of microphthalmia-associated transcription factor (MITF). The regulation of AChE during melanogenesis was hypothesized to be mediated by two transcriptional factors: cAMP responsive element binding protein (CREB) and MITF. In cultured melanoma, application of cAMP suppressed the expression of AChE, as well as the promoter activity of human ACHE gene. This suppression was shown to be mediated by a cAMP responsive element (CRE) located on the ACHE promoter, and mutation of this site eliminated the suppression. In melanoma, over expression of MITF induced the transcription of ACHE gene, and mutation of E-box site of the promoter blocked the induction. In parallel, application of an AChE inhibitor in vitro greatly enhanced acetylcholine-mediated responses of melanogenic gene expressions; but the enhancement was not revealed in the present of agonists of muscarinic acetylcholine receptor. Therefore, our results indicated that AChE transcription is specifically regulated by cAMP-dependent signaling pathway during melanogenesis of B16F10 cells, suggesting a potential role of AChE being played in this differentiation process.

It is usual for information to be unavailable regarding the molecular composition of extracts from herbs or animal tissues that are popular in folk medicine. Here, we present analysis of the alcohol–ether extract from bovine tissue analogous to the basic substance used in such commercial products as Retisin, Imuregen, Actovegin, and Solcoseryl. The tested extract contains a whole spectrum of free amino acids, small proteins and oligopeptides of molecular weight up to 10 kDa, various nucleotides, and a small amount of phospholipids. Among the molecules that can explain some biological activities of the extract were identified those of taurine (2-aminoethanesulfonic acid, a derivative of the amino acid cysteine), several defensins, and bactericidal hemoglobin fragments known as hemocidins. All those molecules identified are natural components of bovine tissues, and a substantial number of them might be biologically active in vivo. Others are sources of readily available nutrients.

Cancer disease has a complicated pathophysiology and is one of the major causes of death and morbidity. Classical cancer therapies include chemotherapy, radiation therapy, and immunotherapy. A typical treatment is chemotherapy, which delivers cytotoxic medications to patients to suppress the uncontrolled growth of cancerous cells. Conventional oral medication has a number of drawbacks, including a lack of selectivity, cytotoxicity, and multi-drug resistance, all of which offer significant obstacles to effective cancer treatment. Multidrug resistance (MDR) remains a major challenge for effective cancer chemotherapeutic interventions. The advent of nanotechnology approach has developed the field of tumor diagnosis and treatment. Cancer nanotechnology enables direct access to tumor cells, resulting in enhanced drug localization and cellular uptake. Since the early 1990’s, several solid lipid nanoparticle (SLN) or SLN-based systems for the delivery of cytotoxic drugs have been manufactured and tested with success. High shear homogenization, microemulsion-based SLN, Supercritical fluid technology, spray drying, and solvent emulsification/evaporation methods can all be used to successfully formulate SLN.There is great potential to enhance cancer chemotherapy by incorporating it into a solid lipid nanoparticle (SLN) drug delivery system. Improving tumor diffusivity, improvement of body distribution, and inhibiting MDR are the main attributes. This type of review article discusses advantages and disadvantages of SLNs, their production techniques, and their potential usage in the treatment of various cancers.

Background: Type 2 diabetes is a common condition that causes the level of sugar (glucose) in the blood to become too high. It can cause symptoms relative insulin deficit, whether due to beta-cell damage, insulin resistance. The study of carnitine and LDH levels in diabetic patients is significant because both play important roles in the metabolism of glucose and fatty acids. Carnitine is a compound that transports fatty acids into the mitochondria for energy production, while LDH (lactate dehydrogenase) is an enzyme involved in the conversion of glucose to lactate. Humans with type 2 diabetes develop lipid accumulation due to carnitine depletion. LDH is an essential physiological molecule in the glycolytic pathway, and its concentration may be indicative of the condition of cellular metabolism.Aim: For measuring and evaluating the levels of serum carnitine and LDH in all study groups.Method: A case-control study was done in the Al-Zahraa Teaching Hospital, Kut, Iraq on 150 Iraqi males and females as patients and control between (April 2022 and January 2023). Their ages ranged between 44 and 77 years. Among them were 120 patients divided into 4 groups 30 type 2 diabetes mellitus; 30 diabetic cardiomyopathies; 30 diabetic nephropathies; 30 diabetic retinopathies and 30 control group where control group's age and gender matched those of the patient groups. All patients gave written informed consent to participate in the clinical study. ELISA was used to measure carnitine and LDH.Result: In present study, it was confirmed that carnitine was significantly lower than the control group and that LDH was significantly higher than the control group. the study demonstrated significant differences in fasting blood sugar and HbA1C levels among the control group, DM2, DCM, DNP, and DRP groups.Conclusion: This case-control study revealed significant differences in carnitine levels, LDH, FBS, and HbA1C levels among patients with Type 2 diabetes mellitus (T2DM) and their complications compared to the control group. These findings suggest alterations in energy metabolism and cellular damage in patients, indicating poorer glycemic control, and supporting the presence of uncontrolled diabetes.

Origanum vulgare (O. Vulgare) is a medicinal and aromatic shrub. It is commonly used as a culinary spice and in "traditional medicine" to cure a variety of diseases. Almost all parts including roots, leaves, stems, and flowers, are employed in medical systems to treat a variety of ailments. In recent scientific literature, among other things, the plant was reported to behave clinical effectiveness against antibacterial, hyperglycemia, anthelminthic and antifungal actions. Carvacrol, limonene, thymol, ocimene, pinene, caryophyllene, terpenes-cymene, and other significant medicinal chemicals have been extracted from this genus through phytochemical studies. The phytochemical ingredients of O. vulgare are highlighted in this review and its traditional uses, as well as evidence-based studies on the plant's diverse pharmacological effects. This illustrates the importance of conducting a thorough investigation in order to report new information on the mechanisms of action of these impacts.

Cancer cells have a special energy metabolism that enables them to multiply quickly. Under normal oxygen conditions, the Warburg effect is a distinguishing aspect of cancer metabolism in which anaerobic glycolysis is favored. Enhanced glycolysis also helps to produce nucleotides, amino acids, lipids, and folic acid, all of which are necessary for cancer cell division. In a variety of metabolic processes, including glycolysis, the co-enzyme nicotinamide adenine dinucleotide (NAD) mediates redox reactions. NAD levels that are higher promote glycolysis and supply energy to cancer cells. NAD metabolism, like energy metabolism, is linked in cancer genesis and could be a potential therapeutic target for cancer treatment. In this research, NAD-consuming enzymes, poly(ADP-ribose) polymerase (PARP) and SIRT1, have been investigated in breast cancer patients, in addition to detect the levels of serum malondialdehyde (MDA), superoxide dismutase (SOD) and vitamin K levels. Sixty participants were enrolled in this study, 30 women with breast cancer and 30 controls. Serum were analysed for determination of the levels of PARP1, SIRT1, MDA, SOD, and vitamin K. The results showed a drop in the expression levels of PARP and that was concomitant with the elevation in the expression levels of SIRT1 and MDA, in addition to the drop in SOD and vitamin K levels. These findings suggest that SIRT1 might be the most NAD-consuming enzyme in cancerous cells rather than PARP (the DNA repair enzyme), and this increase in MDA with the drop in  SOD and vitamin K might be associated with the increase in cell proliferation and a decrease in apoptotic cell death. Finally, this study could be used as a treatment option for patients with breast cancer. could be used as a treatment option for patients with breast cancer.

Background: Cartilage intermediate layer protein 2 (CILP2) is a monomeric glycoprotein that is mostly expressed in the intermediate zone of articular cartilage and can be detected in the extracellular matrix (ECM). The level of CILP2 in serum and its quantity on the articular cartilage surface and throughout the articular cartilage indicates the potential utility of CILP2 for investigation as a biomarker for determining cartilage deterioration in joint disorders. The CILP2 measurement of changes in cartilage biomarkers may be an effective and sensitive tool for detecting the early development of knee osteoarthritis (KOA) in people at risk for KOA. Changes in CILP2 levels may be beneficial for the early diagnosis of KOA, as CILP2 appears to be linked to cartilage thickness reduction in people who are more likely to develop KOA. Platelet rich plasma (PRP) effectiveness of transforming growth factor, platelet-derived growth factor, and the vascular endothelial growth factor is thought to be related to their release. Because of their capacity to increase matrix formation,growth factors have been widely researched for OA and cartilage regeneration. Hyaluronic acid (HA) is a glycosaminoglycan polymer composed of N-acetyl glucosamine and D-glucuronic acid disaccharide molecules. Early embryonic development, inflammatory, wound repair, cell differentiation, and viscoelasticity are all affected by HA, and other biological processes The study was aimed to evaluate effecte of CILP2 levels after intraarticular injecting pure platelet-rich plasma and hyaluronic acid into patients with early knee osteoarthritis.Materials and Methods: BT LAB kits were used to assess the serum CILP2 level. The experimental investigation included 18 control groups, 10 HA injections, and 21 pure PRP injections. age ranged from 35 to 75. The study was exluded all individuals with advanced osteoarthritis in the knee, COVID-19, diabetes, and autoimmune diseases. the time frame running from November 2021 until June 2022. Other variables in our research were age, gender, family history, use of antihypertensive medications or medications for other disorders, and body mass index (BMI). Results: The findings of this study demonstrate elevated CILP2 levels in patients with early KOA before treatments injection compared with the healthy control. After treatments injection, the level of CILP2 was decreased compared with before injection.Conclusions: CILP2 could be one of the prognostic signs of early KOA.

Background: Medical simulation is widely recognized as an essential component of healthcare professional training, including in military training programs around the world. Combat medicine presents unique challenges that can affect the quality of care provided. We aimed to analyze the benefits of medical simulation in the training of combat paramedics.Methods: Two groups of military medics underwent a simulation scenario. One group had previously completed a simulation training with a full-body mannequin, while the second group completed the same training under simulated battle conditions with using imitations of real injuries. The speed and precision of individual treatment procedures, as well as the physiological parameters of the participants, were analyzed during the simulation.Results: A total of 14 participants were included with comparable demographic backgrounds. Both groups had comparable results regarding vital functions and behavior during the evacuation of the wounded as well as the performance tests. The second group performed better in all measured parameters. The rise in salivary cortisol was modest in both groups and no correlation between the Beck anxiety inventory and cortisol rise was found.Conclusion: This pilot study suggests that conducting training in a realistic environment may be important. Participants who underwent realistic training successfully performed all necessary medical interventions.

Background: During pregnancy, COVID-19 viral infection may cause abnormal placental findings. The studies that have focused on the placental structure among infected women with such virus were few. Aim: This study aims to compare the placental histology of women affected by COVID-19 to that of non-infected women.Methods: A prospective case-control study that enrolled placentae of 20 pregnant women who delivered at Al-Khansaa Maternity Teaching Hospital in Ninevah Province /North of Iraq was accomplished. Ten women were positive for CoVID-19 in the antenatal period. Placental biopsies were obtained from the placental maternal side and placental fetal side and were prepared for histopathological examination after staining with (H&E) using a light microscope.Results: This study revealed that the microscopic assessment of placentas women who were diagnosed with COVID-19 exhibited several features in comparison with those of the control group. Among these findings were those of maternal vascular mal perfusion which are represented represented by accelerated villous maturation, thickening of the blood vessel wall, syncytial knots, incomplete villi growth, perivillous fibrin deposition, and congestion of the blood vessels. On the other hand, fetal vascular mal perfusion is represented by delayed villous maturation, increase numbers of large bullous villi with evidence of syncytial injury, reduced vasculosyncytial membrane formation, villous oedema, increase in the nucleated red blood cells' number, cholangitis with intervillous haemorrhage. Placental sections in COVID-19-infected women revealed inflammatory changes that were picked out in both maternal and fetal sides represented by chronic villitis, edematous villi with infiltration with inflammatory cells, and decidual oedema with infiltration with inflam-matory cells. Features of syncytial injury, decidual fibrinoid necrosis, surge in the nucleated red blood cells' counting, infarction, haemorrhage, and calcification were noticed in some sections.Conclusion: This study observed considerable histopathological lesions in the placentae of mothers infected by the COVID-19 virus against the non-infected group.