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1.
Sci Rep ; 10(1): 20987, 2020 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-33268815

RESUMO

Homeostatic control of neuronal excitability by modulation of synaptic inhibition (I) and excitation (E) of the principal neurons is important during brain maturation. The fundamental features of in-utero brain development, including local synaptic E-I ratio and bioenergetics, can be modeled by cerebral organoids (CO) that have exhibited highly regular nested oscillatory network events. Therefore, we evaluated a 'Phase Zero' clinical study platform combining broadband Vis/near-infrared(NIR) spectroscopy and electrophysiology with studying E-I ratio based on the spectral exponent of local field potentials and bioenergetics based on the activity of mitochondrial Cytochrome-C Oxidase (CCO). We found a significant effect of the age of the healthy controls iPSC CO from 23 days to 3 months on the CCO activity (chi-square (2, N = 10) = 20, p = 4.5400e-05), and spectral exponent between 30-50 Hz (chi-square (2, N = 16) = 13.88, p = 0.001). Also, a significant effect of drugs, choline (CHO), idebenone (IDB), R-alpha-lipoic acid plus acetyl-L-carnitine (LCLA), was found on the CCO activity (chi-square (3, N = 10) = 25.44, p = 1.2492e-05), spectral exponent between 1 and 20 Hz (chi-square (3, N = 16) = 43.5, p = 1.9273e-09) and 30-50 Hz (chi-square (3, N = 16) = 23.47, p = 3.2148e-05) in 34 days old CO from schizophrenia (SCZ) patients iPSC. We present the feasibility of a multimodal approach, combining electrophysiology and broadband Vis-NIR spectroscopy, to monitor neurodevelopment in brain organoid models that can complement traditional drug design approaches to test clinically meaningful hypotheses.


Assuntos
Encéfalo/crescimento & desenvolvimento , Organoides/crescimento & desenvolvimento , Acetilcarnitina/farmacologia , Encéfalo/citologia , Encéfalo/efeitos dos fármacos , Encéfalo/fisiologia , Estudos de Casos e Controles , Linhagem Celular , Colina/farmacologia , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Eletrofisiologia , Feminino , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Masculino , Mitocôndrias/metabolismo , Organoides/efeitos dos fármacos , Organoides/fisiologia , Estudo de Prova de Conceito , Esquizofrenia/metabolismo , Espectroscopia de Luz Próxima ao Infravermelho , Ácido Tióctico/farmacologia , Ubiquinona/análogos & derivados , Ubiquinona/farmacologia
2.
Arterioscler Thromb Vasc Biol ; 40(11): 2649-2664, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32938216

RESUMO

OBJECTIVE: Phenotypic switching of vascular smooth muscle cells (VSMCs) plays a critical role in atherosclerosis, vascular restenosis, and hypertension. Choline exerts cardioprotective effects; however, little is known about its effects on VSMC phenotypic switching and vascular remodeling. Here, we investigated whether choline modulates VSMC phenotypic changes and explored the underlying mechanisms. Approach and Results: In cultured VSMCs, choline promoted Nrf2 (nuclear factor erythroid 2-related factor 2) nuclear translocation, inducing the expression of HO-1 (heme oxygenase-1) and NQO-1 (NAD[P]H quinone oxidoreductase-1). Consequently, choline ameliorated Ang II (angiotensin II)-induced increases in NOX (NAD[P]H oxidase) expression and the mitochondrial reactive oxygen species level, thereby attenuating Ang II-induced VSMC phenotypic switching, proliferation, and migration, presumably via M3AChRs (type 3 muscarinic acetylcholine receptors). Downregulation of M3AChR or Nrf2 diminished choline-mediated upregulation of Nrf2, HO-1, and NQO-1 expression, as well as inhibition of VSMC phenotypic transformation, suggesting that M3AChR and Nrf2 activation are responsible for the protective effects of choline. Moreover, activation of the Nrf2 pathway by sulforaphane suppressed Ang II-induced VSMC phenotypic switching and proliferation, indicating that Nrf2 is a key regulator of VSMC phenotypic switching and vascular homeostasis. In a rat model of abdominal aortic constriction in vivo, choline attenuated VSMC phenotypic transformation and vascular remodeling in a manner related to activation of the Nrf2 pathway. CONCLUSIONS: These results reveal that choline impedes VSMC phenotypic switching, proliferation, migration, and vascular remodeling by activating M3AChR and Nrf2-antioxidant signaling and suggest a novel role for Nrf2 in VSMC phenotypic modulation.


Assuntos
Plasticidade Celular/efeitos dos fármacos , Colina/farmacologia , Agonistas Muscarínicos/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Receptor Muscarínico M3/agonistas , Remodelação Vascular/efeitos dos fármacos , Animais , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Masculino , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Fator 2 Relacionado a NF-E2/genética , Fenótipo , Ratos Sprague-Dawley , Receptor Muscarínico M3/genética , Receptor Muscarínico M3/metabolismo , Transdução de Sinais
3.
Artigo em Russo | MEDLINE | ID: mdl-32678562

RESUMO

The analysis of mechanisms of the neuroprotective effect of choline precursors reveals the primary effects of citicoline on the processes of repair of neuronal membranes, a decrease in the degeneration of free fatty acids, and choline alfoscerate, an increase in the production of the neurotransmitter acetylcholine. Although citicoline has a lesser effect on choline secretion than choline alfoscerate, the combination of choline and cytidine in its composition is a universal tool to reduce symptoms of cerebral ischemia, to stabilize cognitive status, superior to the standard benefits of choline. Various mechanisms of the action of citicoline enable to recommend it as a drug effective both in the acute phase of the disease and in the delayed period, giving it the status of a universal nootropic compound.


Assuntos
Colina/farmacologia , Isquemia Encefálica , Citidina Difosfato Colina , Humanos , Fármacos Neuroprotetores , Nootrópicos
4.
Sci Rep ; 10(1): 11996, 2020 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-32686737

RESUMO

α7 nicotinic acetylcholine receptors (nAChRs) are widely expressed in the central nervous system and regarded as potential therapeutic targets for neurodegenerative conditions, such as Alzheimer's disease and schizophrenia. Yet, despite the assumed pathophysiological importance of the α7 nAChR, molecular physiological characterization remains poorly advanced because α7 nAChR cannot be properly folded and sorted to the plasma membranes in most mammalian cell lines, thus preventing the analyses in heterologous expression system. Recently, ER-resident membrane protein NACHO was discovered as a strong chaperone for the functional expression of α7 nAChR in non-permissive cells. Ly6H, a brain-enriched GPI-anchored neurotoxin-like protein, was reported as a novel modulator regulating intracellular trafficking of α7 nAChR. In this study, we established cell lines that stably and robustly express surface α7 nAChR by introducing α7 nAChR, Ric-3, and NACHO cDNA into HEK293 cells (Triple α7 nAChR/RIC-3/NACHO cells; TARO cells), and re-evaluated the function of Ly6H. We report here that Ly6H binds with α7 nAChRs on the cell membrane and modulates the channel activity without affecting intracellular trafficking of α7 nAChR.


Assuntos
Membrana Celular/metabolismo , Ativação do Canal Iônico , Glicoproteínas de Membrana/metabolismo , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Animais , Membrana Celular/efeitos dos fármacos , Galinhas , Colina/farmacologia , Células HEK293 , Humanos , Ativação do Canal Iônico/efeitos dos fármacos , Proteínas Mutantes/metabolismo , Fosfoinositídeo Fosfolipase C/metabolismo , Ligação Proteica/efeitos dos fármacos , Solubilidade
5.
Am J Physiol Lung Cell Mol Physiol ; 319(2): L391-L402, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32640840

RESUMO

Genetic predispositions and environmental exposures are regarded as the main predictors of respiratory disease development. Although the impact of dietary essential nutrient deficiencies on cardiovascular disease, obesity, and type II diabetes has been widely studied, it remains poorly explored in chronic respiratory diseases. Dietary choline and methionine deficiencies are common in the population, and their impact on pulmonary homeostasis is currently unknown. Mice were fed choline- and/or methionine-deficient diets while being exposed to room-air or cigarette smoke for up to 4 wk. Lung functions were assessed using the FlexiVent. Pulmonary transcriptional activity was assessed using gene expression microarrays and quantitative PCR. Immune cells, cytokines, and phosphatidylcholine were quantified in the bronchoalveolar lavage. In this study, we found that short-term dietary choline and/or methionine deficiencies significantly affect lung function in mice in a reversible manner. It also reduced transcriptional levels of collagens and elastin as well as pulmonary surfactant phosphatidylcholine levels. We also found that dietary choline and/or methionine deficiencies markedly interfered with the pulmonary response to cigarette smoke exposure, modulating lung function and dampening inflammation. These findings clearly show that dietary choline and/or methionine deficiencies can have dramatic pathophysiological effects on the lungs and can also affect the pathobiology of cigarette smoke-induced pulmonary alterations. Expanding our knowledge in the field of "nutri-respiratory research" may reveal a crucial role for essential nutrients in pulmonary health and disease, which may prove to be as relevant as genetic predispositions and environmental exposures.


Assuntos
Colina/farmacologia , Homeostase/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Metionina/farmacologia , Fumaça/efeitos adversos , Tabaco/efeitos adversos , Animais , Líquido da Lavagem Broncoalveolar , Citocinas/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Dieta , Feminino , Inflamação/metabolismo , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Surfactantes Pulmonares/metabolismo , Fumar/efeitos adversos
6.
J Agric Food Chem ; 68(26): 6998-7004, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32484692

RESUMO

Naturally occurring sinapine was successfully synthesized through a proline-mediated Knoevenagel-Doebner condensation in ethanol. This synthetic process involving biobased syringaldehyde, Meldrum's acid, and choline chloride offers a sustainable alternative to the existing low-yield pathways. This two-step strategy gives access to sinapine in a 52% overall yield and has been implemented in the synthesis of sinapine analogues, using 4-hydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde, and vanillin as precursors, giving target molecules with 34-61% overall isolated yields. The purity of synthetic sinapine and its analogues (ca. 95%) was assessed by NMR and high-performance liquid chromatography-mass spectrometry analyses. Furthermore, the antioxidant and antimicrobial activities were assessed, and the potential of this series of molecules was confirmed.


Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Antioxidantes/síntese química , Antioxidantes/farmacologia , Colina/análogos & derivados , Antibacterianos/química , Antioxidantes/química , Colina/síntese química , Colina/química , Colina/farmacologia , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Estrutura Molecular
7.
J Dairy Sci ; 103(7): 6070-6086, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32359982

RESUMO

The objective of this experiment was to examine production performance responses to feeding rumen-protected choline (RPC) or methionine (RPM), or both, during the periparturient period. Fifty-four Holstein cows (25 primiparous, 29 multiparous) were used in a randomized block design experiment with a 2 × 2 factorial treatment structure. Cows were blocked by expected calving date and parity and assigned to 1 of 4 treatments: CON (no RPC or RPM); RPC (13.0 g/d of choline ion); RPM (9 g/d of dl-methionine prepartum; 13.5 g/d of dl-methionine postpartum); or RPC + RPM. Treatments were applied once daily as a top-dress from 3 wk before through 5 wk after calving. Dry matter intake and milk production were recorded daily, and milk samples were obtained once weekly. Data were analyzed for primi- and multiparous cows separately, using a repeated-measures mixed model that included random effects of cow and block and fixed effects of RPC, RPM, week, and their interactions; week served as the repeated effect. Initial BW and previous lactation milk yield were included as covariates in the statistical model for multiparous cows. Feeding RPC without RPM increased milk yield for multiparous cows by 8.7 kg/d, but this increase was not observed when RPC was fed with RPM. In multiparous cows, feeding RPM increased milk fat concentration and tended to increase milk fat yield. Because of this, RPM increased fat-corrected milk (FCM) by 2.8 kg/d at wk 2 postpartum, and this increase was sustained through wk 5 postpartum. In contrast, RPM did not affect overall milk fat yield and concentration for primiparous cows. Feeding RPC increased milk yield for primiparous cows by 3.5 kg/d irrespective of RPM inclusion, which is contrary to observations in multiparous cows, where RPC increased milk yield only in the absence of RPM. These results indicate that responses to RPC during the periparturient period may be dependent upon supply of methionine. Our observations also demonstrate that primi- and multiparous cows respond differently to RPC and RPM supplemented individually or simultaneously during the periparturient period. This variation in response could have been mediated by putative differences in choline and methionine requirements of primiparous versus multiparous cows, or by differences in the levels of milk production between the 2 groups (36 vs. 25 kg of FCM/d). However, cows in this study did not experience severe negative energy balance (mean nadirs of -6.6 and -5.0 Mcal/d for multiparous and primiparous cows, respectively), which likely affected their responses to RPC and RPM.


Assuntos
Colina/farmacologia , Suplementos Nutricionais , Lactação/efeitos dos fármacos , Metionina/farmacologia , Período Periparto/efeitos dos fármacos , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Bovinos , Colina/administração & dosagem , Dieta/veterinária , Metabolismo Energético/fisiologia , Feminino , Lactação/fisiologia , Fenômenos Fisiológicos da Nutrição Materna , Metionina/administração & dosagem , Leite/química , Paridade , Gravidez , Rúmen/metabolismo
8.
J Nutr ; 150(7): 1958-1965, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32271922

RESUMO

BACKGROUND: Buttermilk contains a mixture of choline forms; it is high in phosphatidylcholine (PC) and sphingomyelin (SM), which could have an impact on immune system development and function. OBJECTIVES: We aimed to determine the effect of feeding buttermilk-derived choline forms during pregnancy and lactation on maternal immune function. METHODS: Sprague Dawley dams (n = 8 per diet) were randomly assigned midway through pregnancy (10 d of gestation) to 1 of 3 experimental diets, containing 1.7 g/kg choline: control [100% free choline (FC)]; buttermilk [37% PC, 34% SM, 17% glycerophosphocholine (GPC), 7% FC, 5% phosphocholine]; or placebo (50% PC, 25% FC, 25% GPC). Dams consumed the same diet until the end of the lactation period (21 d after parturition). Cell phenotypes and cytokine production by mitogen-stimulated splenocytes were measured and compared using 1-factor ANOVA test in order to asses the effect of diet on immune fuction of lactating dams (main outcome). RESULTS: After ConA stimulation, splenocytes from dams in the buttermilk group produced more IL-2 (30%), TNF-α (30%), and IFN-γ (42%) compared with both the placebo and control diets. Placebo-fed dams had a higher proportion of CD8+ cells expressing CD152+ (22%) in spleen, and splenocytes from dams that were fed the buttermilk and the placebo diets produced about 50% and 53% more IL-10 after LPS and OVA stimulation, respectively, compared with the control group. CONCLUSIONS: Feeding buttermilk-derived choline forms during pregnancy and lactation had a beneficial impact on the immune system of Sprague Dawley rat dams, especially on T-cell function.


Assuntos
Leitelho/análise , Colina/análise , Colina/farmacologia , Fenômenos Fisiológicos da Nutrição Materna , Linfócitos T/efeitos dos fármacos , Linfócitos T/fisiologia , Ração Animal/análise , Animais , Concanavalina A/farmacologia , Dieta/veterinária , Feminino , Gravidez , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley
9.
Arterioscler Thromb Vasc Biol ; 40(5): 1239-1255, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32212854

RESUMO

OBJECTIVE: Gut microbial metabolism of dietary choline, a nutrient abundant in a Western diet, produces trimethylamine (TMA) and the atherothrombosis- and fibrosis-promoting metabolite TMA-N-oxide (TMAO). Recent clinical and animal studies reveal that elevated TMAO levels are associated with heightened risks for both cardiovascular disease and incident chronic kidney disease development. Despite this, studies focusing on therapeutically targeting gut microbiota-dependent TMAO production and its impact on preserving renal function are limited. Approach and Results: Herein we examined the impact of pharmacological inhibition of choline diet-induced gut microbiota-dependent production of TMA, and consequently TMAO, on renal tubulointerstitial fibrosis and functional impairment in a model of chronic kidney disease. Initial studies with a gut microbial choline TMA-lyase mechanism-based inhibitor, iodomethylcholine, confirmed both marked suppression of TMA generation, and consequently TMAO levels, and selective targeting of the gut microbial compartment (ie, both accumulation of the drug in intestinal microbes and limited systemic exposure in the host). Dietary supplementation of either choline or TMAO significantly augmented multiple indices of renal functional impairment and fibrosis associated with chronic subcutaneous infusion of isoproterenol. However, the presence of the gut microbiota-targeting inhibitor iodomethylcholine blocked choline diet-induced elevation in TMAO, and both significantly improved decline in renal function, and significantly attenuated multiple indices of tubulointerstitial fibrosis. Iodomethylcholine treatment also reversed many choline diet-induced changes in cecal microbial community composition associated with TMAO and renal functional impairment. CONCLUSIONS: Selective targeting of gut microbiota-dependent TMAO generation may prevent adverse renal structural and functional alterations in subjects at risk for chronic kidney disease.


Assuntos
Bactérias/efeitos dos fármacos , Proteínas de Bactérias/antagonistas & inibidores , Colina/farmacologia , Inibidores Enzimáticos/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacos , Rim/efeitos dos fármacos , Liases/antagonistas & inibidores , Metilaminas/metabolismo , Insuficiência Renal Crônica/tratamento farmacológico , Animais , Bactérias/enzimologia , Proteínas de Bactérias/metabolismo , Colina/análogos & derivados , Modelos Animais de Doenças , Fibrose , Rim/metabolismo , Rim/patologia , Rim/fisiopatologia , Liases/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Insuficiência Renal Crônica/metabolismo , Insuficiência Renal Crônica/microbiologia , Insuficiência Renal Crônica/patologia
10.
J Dairy Sci ; 103(5): 4174-4191, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32171515

RESUMO

Objectives were to evaluate the effects of altering timing of initiating and duration of supplementing rumen-protected choline (RPC) on lactation performance in dairy cows. The hypothesis was that RPC increases yields of milk and milk components, regardless of when supplementation is initiated, and that the effects of supplementing RPC starting prepartum and continuing post-transition would be additive. Cows at 241 ± 2.2 d of gestation were blocked by parity group (49 entering lactation 2, 50 entering lactation >2) and 305-d milk yield and, within block, assigned randomly to 1 of 4 treatments arranged as a 2 × 2 factorial with 2 levels of choline in transition, from 21 d pre- to 21 d postpartum, and 2 levels of choline in post-transition, from 22 to 105 d postpartum. The 2 levels of RPC supplemented were either 0 g/d or 12.9 g/d of choline ion fed as 60 g/d of an RPC product that was top-dressed onto the total mixed ration. Thus, treatments were as follows: NN (n = 25): no choline in transition or post-transition; NC (n = 25): no choline in transition and choline in post-transition; CN (n = 25): choline in transition and no choline in post-transition; CC (n = 24): choline in transition and in post-transition. Prepartum, treatments were supplemented (mean ± SD) for the last 18.8 ± 5.7 and 19.2 ± 5.0 d of gestation in treatments with 0 or 12.9 g/d of choline ion, respectively. Supplementing RPC prepartum did not affect dry matter intake (DMI), body weight (BW), or body condition score (BCS) in the last 3 weeks of gestation. Likewise, RPC did not affect the yield or the composition of colostrum. Supplementation with RPC during transition increased fat percent by 0.02 percentage units, fat yield by 0.16 kg/d, and energy-corrected milk (ECM) by 3.1 kg/d in the first 21 d postpartum, and increased fat yield by 0.10 kg/d and ECM by 2.4 kg/d from 22 to 105 d postpartum. Supplementing RPC during transition did not affect DMI postpartum, but it improved feed efficiency, and cows produced 0.11 kg/d more ECM per kg of DMI. Changes in BW and BCS during the first 21 d postpartum did not differ between treatments. Cows fed RPC during transition had more negative net energy balance and 0.1 unit smaller BCS in the first 105 d postpartum than non-supplemented cows. Supplementing RPC in post-transition did not influence productive performance in dairy cows, and choline supplementation during transition or post-transition did not affect measures of reproduction. Collectively, supplementing RPC to supply 12.9 g/d of choline ion benefited productive performance in dairy cows when supplementation occurred during the transition period, but no additional benefit was observed from supplementing RPC past 22 d postpartum.


Assuntos
Bovinos , Colina/farmacologia , Dieta/veterinária , Lactação , Animais , Peso Corporal , Colina/administração & dosagem , Indústria de Laticínios , Suplementos Nutricionais , Metabolismo Energético , Feminino , Lactação/efeitos dos fármacos , Leite , Período Pós-Parto , Rúmen/metabolismo
11.
Biol Pharm Bull ; 43(3): 463-473, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32115504

RESUMO

Choline as a quaternary amine nutrient is metabolized to trimethylamine by gut microbiota and subsequently oxidized to circulating trimethylamine-N-oxide (TMAO), a gut-derived metabolite associated with liver toxicity and cardiovascular disease. The study was to probe the possible vasoprotective and hepatoprotective effects of total saponins of Gynostemma pentaphyllum (TSGP) in 3% high-choline water-feeding mice. The purified TSGP was obtained with content of 83.0% saponins, and its antioxidant activities were evaluated in vitro. Furthermore, the mice fed with high choline for 8 weeks significantly expressed vascular endothelial dysfunction and liver oxidative stress (p < 0.01 vs. Normal). Administration of TSGP at 400 and 800 mg/kg·body weight (b.w.) significantly lowered the serum total cholesterol (TC), triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), endothelin-1 (ET-1) and thromboxane A2 (TXA2) levels, as well as hepatic malondialdehyde (MDA) formation, but effectively elevated the serum nitric oxide (NO), endothelial nitric oxide synthase (eNOS) and prostaglandin I2 (PGI2) levels, as well as alanine aminotransferase (ALT), aspartate aminotransferase (AST), T-superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in high choline-fed mice. Hematoxylin-eosin (H&E) and oil red O staining also suggested that TSGP could exert the significant protection against endothelial dysfunction and liver injury in high choline-treated mice. These findings suggest that TSGP is of the saponins-enriched extract, and is a good candidate of dietary supplement and therapeutic application in vascular and hepatic oxidative injury.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Colina/farmacologia , Gynostemma , Extratos Vegetais/farmacologia , Saponinas/farmacologia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Peso Corporal/efeitos dos fármacos , LDL-Colesterol/sangue , Endotélio Vascular/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Fígado/metabolismo , Fígado/patologia , Malondialdeído/metabolismo , Metilaminas , Camundongos , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Tromboxano A2/sangue , Triglicerídeos/sangue
12.
Toxicol Lett ; 321: 21-31, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31830555

RESUMO

Nerve agents inhibit acetylcholinesterase (AChE), leading to a build-up of acetylcholine (ACh) and overstimulation at cholinergic synapses. Current post-exposure nerve agent treatment includes atropine to treat overstimulation at muscarinic synapses, a benzodiazepine anti-convulsant, and an oxime to restore the function of AChE. Aside from the oxime, the components do not act directly to reduce the overstimulation at nicotinic synapses. The false transmitters acetylmonoethylcholine (AMECh) and acetyldiethylcholine (ADECh) are analogs of ACh, synthesised similarly at synapses. AMECh and ADECh are partial agonists, with reduced activity compared to ACh, so it was hypothesised the false transmitters could reduce overstimulation. Synthetic routes to AMECh and ADECh, and their precursors, monoethylcholine (MECh) and diethylcholine (DECh), were devised, allowing them to be produced easily on a laboratory-scale. The mechanism of action of the false transmitters was investigated in vitro. AMECh acted as a partial agonist at human muscarinic (M1 and M3) and muscle-type nicotinic receptors, and ADECh was a partial agonist only at certain muscarinic subtypes. Their precursors acted as antagonists at muscle-type nicotinic, but not muscarinic receptors. Administration of MECh and DECh improved neuromuscular function in the soman-exposed guinea-pig hemi-diaphragm preparation. False transmitters may therefore help reduce nerve agent induced overstimulation at cholinergic synapses.


Assuntos
Acetilcolina/análogos & derivados , Antídotos/farmacologia , Colina/análogos & derivados , Inibidores da Colinesterase/envenenamento , Diafragma/inervação , Agentes Neurotóxicos/envenenamento , Neurotransmissores/farmacologia , Intoxicação por Organofosfatos/tratamento farmacológico , Soman/envenenamento , Sinapses/efeitos dos fármacos , Acetilcolina/síntese química , Acetilcolina/metabolismo , Acetilcolina/farmacologia , Acetilcolinesterase/metabolismo , Animais , Antídotos/síntese química , Células CHO , Linhagem Celular Tumoral , Colina/síntese química , Colina/farmacologia , Cricetulus , Agonismo Parcial de Drogas , Cobaias , Humanos , Masculino , Neurotransmissores/síntese química , Intoxicação por Organofosfatos/enzimologia , Intoxicação por Organofosfatos/fisiopatologia , Receptores Colinérgicos/efeitos dos fármacos , Receptores Colinérgicos/genética , Receptores Colinérgicos/metabolismo , Sinapses/enzimologia
13.
Artigo em Inglês | MEDLINE | ID: mdl-31493584

RESUMO

Aflatoxin B1 (AFB1) is one of the most important mycotoxins due to its hepatotoxic and carcinogenic effects on animals. The effect of dietary supplementation with vegetable choline (VC) at 400, 800, and 1200 mg/kg against the deleterious effects of AFB1 (2 ppm/kg diet) in the liver of Nile tilapia (Oreochromis niloticus) was studied. The experimental period was 81 days, and the diet with VC was offered to the fish for 60 days prior to challenge with AFB1. Diets with AFB1 were tested in three replications and animals were analyzed at days 14 and 21 of dietary intake. The addition of VC to tilapia diet increased body weight (days 30 and 60 pre-challenge and day 21 post-challenge). The group fed aflatoxin-contaminated diet presented significantly reduced antioxidant enzymes and increased reactive oxygen species (ROS) levels, thiobarbituric acid reactive species (TBARS) levels, and protein carbonyl (PC) content in the liver. Dietary supplementation with VC at 800 and 1200 mg/kg demonstrated a significant protective effect, avoiding the increase of ROS, TBARS, and PC levels in the liver of tilapia from the aflatoxin contaminated groups. Thus, dietary VC supplementation may be used in tilapia to increase antioxidant status and reduce the negative effects caused by AFB1 toxicity. Based on the findings, it is recommended to use VC as a food supplement for Nile tilapia in order to avoid AFB1 toxication. In addition, decreased aflatoxin toxicity can be attributed to the VC antioxidant property.


Assuntos
Aflatoxina B1/toxicidade , Ração Animal/análise , Colina/farmacologia , Ciclídeos , Doenças dos Peixes/induzido quimicamente , Contaminação de Alimentos , Aflatoxina B1/administração & dosagem , Animais , Catalase/genética , Catalase/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Doença Hepática Induzida por Substâncias e Drogas/veterinária , Colina/administração & dosagem , Dieta/veterinária , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Doenças dos Peixes/tratamento farmacológico , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo
14.
Brain ; 143(1): 94-111, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31855247

RESUMO

Cerebral choline metabolism is crucial for normal brain function, and its homoeostasis depends on carrier-mediated transport. Here, we report on four individuals from three families with neurodegenerative disease and homozygous frameshift mutations (Asp517Metfs*19, Ser126Metfs*8, and Lys90Metfs*18) in the SLC44A1 gene encoding choline transporter-like protein 1. Clinical features included progressive ataxia, tremor, cognitive decline, dysphagia, optic atrophy, dysarthria, as well as urinary and bowel incontinence. Brain MRI demonstrated cerebellar atrophy and leukoencephalopathy. Moreover, low signal intensity in globus pallidus with hyperintensive streaking and low signal intensity in substantia nigra were seen in two individuals. The Asp517Metfs*19 and Ser126Metfs*8 fibroblasts were structurally and functionally indistinguishable. The most prominent ultrastructural changes of the mutant fibroblasts were reduced presence of free ribosomes, the appearance of elongated endoplasmic reticulum and strikingly increased number of mitochondria and small vesicles. When chronically treated with choline, those characteristics disappeared and mutant ultrastructure resembled healthy control cells. Functional analysis revealed diminished choline transport yet the membrane phosphatidylcholine content remained unchanged. As part of the mechanism to preserve choline and phosphatidylcholine, choline transporter deficiency was implicated in impaired membrane homeostasis of other phospholipids. Choline treatments could restore the membrane lipids, repair cellular organelles and protect mutant cells from acute iron overload. In conclusion, we describe a novel childhood-onset neurometabolic disease caused by choline transporter deficiency with autosomal recessive inheritance.


Assuntos
Antígenos CD/genética , Transtornos Heredodegenerativos do Sistema Nervoso/genética , Proteínas de Transporte de Cátions Orgânicos/genética , Adolescente , Ataxia/genética , Ataxia/fisiopatologia , Atrofia , Cerebelo/diagnóstico por imagem , Cerebelo/patologia , Colina/farmacologia , Disfunção Cognitiva/genética , Disfunção Cognitiva/fisiopatologia , Vesículas Citoplasmáticas/efeitos dos fármacos , Vesículas Citoplasmáticas/ultraestrutura , Transtornos de Deglutição/genética , Transtornos de Deglutição/fisiopatologia , Disartria/genética , Disartria/fisiopatologia , Retículo Endoplasmático/efeitos dos fármacos , Retículo Endoplasmático/ultraestrutura , Incontinência Fecal/genética , Incontinência Fecal/fisiopatologia , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/ultraestrutura , Mutação da Fase de Leitura , Globo Pálido/diagnóstico por imagem , Transtornos Heredodegenerativos do Sistema Nervoso/diagnóstico por imagem , Transtornos Heredodegenerativos do Sistema Nervoso/patologia , Transtornos Heredodegenerativos do Sistema Nervoso/fisiopatologia , Homozigoto , Humanos , Leucoencefalopatias/diagnóstico por imagem , Leucoencefalopatias/genética , Leucoencefalopatias/fisiopatologia , Imagem por Ressonância Magnética , Masculino , Microscopia Eletrônica , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/ultraestrutura , Nootrópicos/farmacologia , Atrofia Óptica/genética , Atrofia Óptica/fisiopatologia , Linhagem , Ribossomos/efeitos dos fármacos , Ribossomos/ultraestrutura , Substância Negra/diagnóstico por imagem , Síndrome , Tremor/genética , Tremor/fisiopatologia , Incontinência Urinária/genética , Incontinência Urinária/fisiopatologia
15.
PLoS One ; 14(9): e0222211, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31527873

RESUMO

Choline geranate (also described as Choline And GEranic acid, or CAGE) has been developed as a novel biocompatible antiseptic material capable of penetrating skin and aiding the transdermal delivery of co-administered antibiotics. The antibacterial properties of CAGE were analyzed against 24 and 72 hour old biofilms of 11 clinically isolated ESKAPE pathogens (defined as Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Enterobacter sp, respectively), including multidrug resistant (MDR) isolates. CAGE was observed to eradicate in vitro biofilms at concentrations as low as 3.56 mM (0.156% v:v) in as little as 2 hours, which represents both an improved potency and rate of biofilm eradication relative to that reported for most common standard-of-care topical antiseptics in current use. In vitro time-kill studies on 24 hour old Staphylococcus aureus biofilms indicate that CAGE exerts its antibacterial effect upon contact and a 0.1% v:v solution reduced biofilm viability by over three orders of magnitude (a 3log10 reduction) in 15 minutes. Furthermore, disruption of the protective layer of exopolymeric substances in mature biofilms of Staphylococcus aureus by CAGE (0.1% v:v) was observed in 120 minutes. Insight into the mechanism of action of CAGE was provided with molecular modeling studies alongside in vitro antibiofilm assays. The geranate ion and geranic acid components of CAGE are predicted to act in concert to integrate into bacterial membranes, affect membrane thinning and perturb membrane homeostasis. Taken together, our results show that CAGE demonstrates all properties required of an effective topical antiseptic and the data also provides insight into how its observed antibiofilm properties may manifest.


Assuntos
Anti-Infecciosos Locais/farmacologia , Colina/farmacologia , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Testes de Sensibilidade Microbiana/métodos
16.
Pak J Pharm Sci ; 32(3 (Supplementary)): 1193-1199, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31303590

RESUMO

Cognitive behavior is associated with physiological processes that affect the working performance of an individual. Cognitive control is used to override self-serving impulses and behave in socially desirable manner. The objective of the study is to compare the effects of Choline with Fluoxetine and Clozapine for the modulation of cognitive behavior including learning, memory, locomotor, exploratory behavior and anxiety. The study was based on twenty four albino rats divided into four equal groups: (1) Control kept on normal saline (2) Fluoxetine (3) Clozapine (4) Choline. Morris Water Maze (WM) test was used for the psychological assessment based on neural mechanism involved in spatial learning and memory. Open field activity test evaluated locomotor and exploratory behavior. The behavior modulation in WM test and open field activity test was determined at 1st, 3rd, 5th and 7th week. Fluoxetine, Clozapine and Choline were used as drugs and administered to the rat groups mentioned earlier. The modulation of behavior in WM test and Open field activity test was recorded at 1st, 3rd, 5th and 7th week after administering the drugs. Impairment in learning behavior in Fluoxetine treated group was observed at 1st, 3rd, 5th and 7th week and in Clozapine group at 1st and 2nd week when compared to Control (Saline) group. Rise in latency time was observed in Fluoxetine treated group but was not significant. Clozapine and Choline had exhibited beneficial effects in memory retention and prevention of learning impairment. The findings have led to the conclusion that Choline and Clozapine improve the memory retention after continuous administration of 5 and 7 weeks. Moreover, Clozapine has different receptor specificity as compared to Choline. However, both improve the learning capability and enhance the memory in rats. Meanwhile, Fluoxetine did not show any considerable enhancement of memory.


Assuntos
Colina/farmacologia , Clozapina/farmacologia , Cognição/efeitos dos fármacos , Fluoxetina/farmacologia , Animais , Antipsicóticos/farmacologia , Comportamento Exploratório/efeitos dos fármacos , Aprendizagem/efeitos dos fármacos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Memória/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Ratos
17.
New Phytol ; 224(1): 258-273, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31246280

RESUMO

The continuous growth of roots requires the balance between cell division and differentiation. Reactive oxygen species (ROS) and auxin are important regulators of root development by affecting cell division and differentiation. The mechanism controlling the coordination of cell division and differentiation is not well understood. Using a forward genetic screen, we isolated a mutant, defective primary root 2 (dpr2), defective in root apical meristem (RAM) maintenance. The DPR2 gene encodes phosphoethanolamine N-methyltransferase 1 (PEAMT1) that catalyzes phosphocholine biosynthesis in Arabidopsis. We characterized the primary root phenotypes of dpr2 using various marker lines, using histochemical and pharmacological analysis to probe early root development. Loss-of-function of DPR2/PEAMT1 resulted in RAM consumption by affecting root stem cell niche, division zone, elongation and differentiation zone (EDZ). PIN-FORMED (PIN) protein abundance, PIN2 polar distribution and general endocytosis were impaired in the root tip of dpr2. Excess hydrogen peroxide and auxin accumulate in the EDZ of dpr2, leading to RAM consumption by accelerating cell differentiation. Suppression of ROS over-accumulation or inhibition of auxin signalling partially prevent RAM differentiation in dpr2 after choline starvation. Taken together, we conclude that the EDZ of the root tip is most sensitive to choline shortage, leading to RAM consumption through an ROS-auxin regulation module.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Arabidopsis/enzimologia , Diferenciação Celular/efeitos dos fármacos , Ácidos Indolacéticos/farmacologia , Meristema/citologia , Metiltransferases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/genética , Divisão Celular/efeitos dos fármacos , Colina/farmacologia , Endocitose/efeitos dos fármacos , Etanolaminas/metabolismo , Meristema/efeitos dos fármacos , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Mutação/genética , Oniocompostos/farmacologia , Fenótipo , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo
18.
Arterioscler Thromb Vasc Biol ; 39(6): 1045-1054, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31070450

RESUMO

Objective- FMO (flavin-containing monooxygenase) 3 converts bacterial-derived trimethylamine to trimethylamine N-oxide (TMAO), an independent risk factor for cardiovascular disease. We generated FMO3 knockout (FMO3KO) mouse to study its effects on plasma TMAO, lipids, glucose/insulin metabolism, thrombosis, and atherosclerosis. Approach and Results- Previous studies with an antisense oligonucleotide (ASO) knockdown strategy targeting FMO3 in LDLRKO (low-density lipoprotein receptor knockout) mice resulted in major reductions in TMAO levels and atherosclerosis, but also showed effects on plasma lipids, insulin, and glucose. Although FMO3KO mice generated via CRISPR/Cas9 technology bred onto the LDLRKO background did exhibit similar effects on TMAO levels, the effects on lipid metabolism were not as pronounced as with the ASO knockdown model. These differences could result from either off-target effects of the ASO or from a developmental adaptation to the FMO3 deficiency. To distinguish these possibilities, we treated wild-type and FMO3KO mice with control or FMO3 ASOs. FMO3-ASO treatment led to the same extent of lipid-lowering effects in the FMO3KO mice as the wild-type mice, indicating off-target effects. The levels of TMAO in LDLRKO mice fed an atherogenic diet are very low in both wild-type and FMO3KO mice, and no significant effect was observed on atherosclerosis. When FMO3KO and wild-type mice were maintained on a 0.5% choline diet, FMO3KO showed a marked reduction in both TMAO and in vivo thrombosis potential. Conclusions- FMO3KO markedly reduces systemic TMAO levels and thrombosis potential. However, the previously observed large effects of an FMO3 ASO on plasma lipid levels appear to be due partly to off-target effects.


Assuntos
Aterosclerose/metabolismo , Colina/metabolismo , Metilaminas/metabolismo , Oxigenases/genética , Trombose/metabolismo , Animais , Aterosclerose/genética , Colina/farmacologia , Modelos Animais de Doenças , Metabolismo dos Lipídeos/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Oxigenases/metabolismo , Reação em Cadeia da Polimerase/métodos , Distribuição Aleatória , Valores de Referência , Trombose/fisiopatologia
19.
PLoS One ; 14(5): e0217160, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31141525

RESUMO

Choline and methionine serve essential roles in the liver that may interact with glucose metabolism. Our objectives were to quantify glucose export, cellular glycogen, and expression of genes controlling oxidation and gluconeogenesis in primary bovine neonatal hepatocytes exposed to increasing concentrations of choline chloride (CC) and D,L-methionine (DLM) with or without fatty acids (FA). Primary hepatocytes isolated from 3 Holstein calves were maintained as monolayer cultures for 24 h before treatment with CC (61, 128, 2028, 4528 µmol/L) and DLM (16, 30, 100, 300 µmol/L) with or without a 1 mmol/L FA cocktail in a factorial design. After 24 h, media was harvested to quantify glucose, ß-hydroxybutyrate (BHB), and cells harvested to quantify glycogen, DNA, and gene expression. No interactions between CC and DLM were detected. The potential two-way interaction between CC or DLM and FA was partitioned into three contrasts when P ≤ 0.20: linear without FA, linear with FA, difference of slope. Fatty acids did not affect glucose or cellular glycogen but increased pyruvate carboxylase (PC), cytosolic and mitochondrial phosphoenolpyruvate carboxykinase (PEPCKc, PEPCKm), and glucose-6-phosphatase (G6PC) expression. Increasing CC decreased glucose but increased cellular glycogen. Expression of PC and PEPCKc was increased by CC during FA treatment. Increasing DLM did not affect metabolites or PC expression, although PEPCKc was marginally decreased. Methionine did not affect G6PC, while CC had a marginal quadratic effect on G6PC. Oxidative and gluconeogenic enzymes appear to respond to FA in primary bovine neonatal hepatocytes. Increased PC and PEPCKc by CC during FA treatment suggest increased gluconeogenic capacity. Changes in G6PC may have shifted glucose-6-phosphate towards cellular glycogen; however, subsequent examination of G6PC protein is needed. Unaltered PC and marginally decreased PEPCKc suggest increased oxidative capacity with DLM, although BHB export was unaltered. The differential regulation supports unique effects of CC and DLM within bovine hepatocytes.


Assuntos
Colina/farmacologia , Ácidos Graxos/farmacologia , Gluconeogênese/efeitos dos fármacos , Glucose/metabolismo , Hepatócitos/metabolismo , Metionina/farmacologia , Animais , Animais Recém-Nascidos , Bovinos , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos
20.
Poult Sci ; 98(8): 3304-3312, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-30941414

RESUMO

Five hundred and forty 19-wk-old HyLine Brown laying hens were randomly distributed to 6 dietary treatments and fed 1of 6 corn-soybean meal-based diets added into choline with 0, 425, 850, 1,700, 3,400, and 6,800 mg/kg to investigate effects of dietary choline supplementation on lipid profiles of egg yolk, serum and liver, and hepatic redox status of laying hens. Yolk weight and total lipid, triglyceride, cholesterol and phosphatidylcholine, serum triglyceride, cholesterol, apolipoprotein B 100 (apoB 100), and very low density lipoprotein (VLDL), and liver relative weight, total lipid, triglyceride and apoB 100 as well as hepatic total superoxide dismutase and glutathione peroxidase (GSH-Px) activities, total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in laying hens at weeks 58 and 68 of age were determined. The differences (P < 0.001) were caused by choline treatments in yolk phosphatidylcholine (at 850 mg/kg or more choline), serum VLDL, and liver triglyceride (at 1,700 and 3,400 mg/kg choline) of birds, at weeks 58 and 68 of age, and yolk total lipids were elevated (P < 0.05) by supplemental choline at 3,400 mg/kg whereas liver total lipids were reduced (P < 0.05) by 1,700 and 3,400 mg/kg choline addition. Hens fed diets supplemented choline had higher (P = 0.005) liver GSH-Px activity (with 3,400 mg/kg choline) and greater (P = 0.014) T-AOC (with 1,700 mg/kg choline) than those fed diets with 0 and 425 mg/kg choline addition. Choline affected serum VLDL, liver total lipid, triglyceride and apoB 100 at weeks 58 and 68 of age and hepatic GSH-Px activity, T-AOC and MDA at week 68 of age quadratically (P < 0.05), whereas it influenced total lipid and phosphatidylcholine of egg yolk linearly (P < 0.05) and quadratically (P < 0.05). In conclusion, dietary choline supplementation elevated yolk total lipid and phosphatidylcholine and serum VLDL, reduced liver total lipid and triglyceride, and enhanced hepatic GSH-Px activity and T-AOC in laying hens.


Assuntos
Ração Animal/análise , Colina/farmacologia , Gema de Ovo/química , Lipídeos/sangue , Animais , Galinhas , Dieta/veterinária , Feminino , Glutationa Peroxidase/análise , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/enzimologia , Fígado/metabolismo , Oxirredução/efeitos dos fármacos , Fosfatidilcolinas/análise , Distribuição Aleatória , Superóxido Dismutase/análise
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