Assessment of Cholinesterase inhibition activity of birds inhabiting pesticide exposed croplands and protected area in hot semi-arid region of Pakistan / Avaliação da atividade de inibição da colinesterase de pássaros que habitam áreas cultivadas e áreas protegidas expostas a pesticidas na região semiárida quente do Paquistão

Abstract Acetylcholinesterase (AChE) activity levels can be used as an indicator for AChE inhibition due to pesticide poisoning in bird species. We assessed the comparative brain cholinesterase (AChE) activity level of five bird species inhabiting pesticide exposed croplands and Protected Area i.e. Deva Vatala National Park (DVNP), Bhimber by using a spectrophotometric method. AChE activity levels ranged from 56.3 to 85.9 µmol/min/g of brain tissue of birds representing DVNP. However, AChE activity levels ranged from 27.6 to 79.9 µmol/min/g of brain tissue of birds representing croplands. AChE activity levels observed in Jungle babbler, Common babbler, and Red-vented bulbul showed significant differences (P < 0.05) at two sites. However, White wagtail and Black drongo demonstrated non-significant differences (P > 0.05). Maximum inhibition was recorded in Jungle babbler (53%) followed by Common babbler (35%), Red-vented bulbul (18%), White wagtail (15%), and Black drongo (7%). The brain cholinesterase inhibition levels under-protected ecosystems (DVNP, Bhimber) and agricultural landscape suggest insecticidal contamination and its impact on avifauna diversity. The study also emphasizes on the importance of pesticide-free zones to protect the biodiversity of birds.

Resumo Os níveis de atividade da acetilcolinesterase (AChE) podem ser usados ​​como um indicador para a inibição da AChE devido ao envenenamento por pesticidas em espécies de aves. Avaliamos o nível de atividade comparativa da colinesterase cerebral (AChE) de cinco espécies de aves que habitam áreas cultivadas expostas a pesticidas e Área Protegida, ou seja, Deva Vatala National Park (DVNP), Bhimber, usando um método espectrofotométrico. Os níveis de atividade da AChE variaram de 56,3 a 85,9 µmol / min / g de tecido cerebral de aves representando DVNP. No entanto, os níveis de atividade da AChE variaram de 27,6 a 79,9 µmol / min / g de tecido cerebral de aves representando áreas de cultivo. Os níveis de atividade de AChE observados no tagarela da selva, tagarela comum e bulbul vermelho exalado mostraram diferenças significativas (P < 0,05) em dois locais. No entanto, alvéola branca e drongo preto demonstraram diferenças não significativas (P > 0,05). A inibição máxima foi registrada no tagarela da selva (53%), seguido pelo tagarela comum (35%), bulbul vermelho (18%), alvéola branca (15%) e drongo preto (7%). Os níveis de inibição da colinesterase cerebral nos ecossistemas subprotegidos (DVNP, Bhimber) e na paisagem agrícola sugerem contaminação por inseticida e seu impacto na diversidade da avifauna. O estudo também enfatiza a importância das zonas livres de pesticidas para proteger a biodiversidade das aves.

Assessment of Cholinesterase inhibition activity of birds inhabiting pesticide exposed croplands and protected area in hot semi-arid region of Pakistan / Avaliação da atividade de inibição da colinesterase de pássaros que habitam áreas cultivadas e áreas protegidas expostas a pesticidas na região semiárida quente do Paquistão

Acetylcholinesterase (AChE) activity levels can be used as an indicator for AChE inhibition due to pesticide poisoning in bird species. We assessed the comparative brain cholinesterase (AChE) activity level of five bird species inhabiting pesticide exposed croplands and Protected Area i.e. Deva Vatala National Park (DVNP), Bhimber by using a spectrophotometric method. AChE activity levels ranged from 56.3 to 85.9 µmol/min/g of brain tissue of birds representing DVNP. However, AChE activity levels ranged from 27.6 to 79.9 µmol/min/g of brain tissue of birds representing croplands. AChE activity levels observed in Jungle babbler, Common babbler, and Red-vented bulbul showed significant differences (P 0.05). Maximum inhibition was recorded in Jungle babbler (53%) followed by Common babbler (35%), Red-vented bulbul (18%), White wagtail (15%), and Black drongo (7%). The brain cholinesterase inhibition levels under-protected ecosystems (DVNP, Bhimber) and agricultural landscape suggest insecticidal contamination and its impact on avifauna diversity. The study also emphasizes on the importance of pesticide-free zones to protect the biodiversity of birds.

Os níveis de atividade da acetilcolinesterase (AChE) podem ser usados como um indicador para a inibição da AChE devido ao envenenamento por pesticidas em espécies de aves. Avaliamos o nível de atividade comparativa da colinesterase cerebral (AChE) de cinco espécies de aves que habitam áreas cultivadas expostas a pesticidas e Área Protegida, ou seja, Deva Vatala National Park (DVNP), Bhimber, usando um método espectrofotométrico. Os níveis de atividade da AChE variaram de 56,3 a 85,9 µmol / min / g de tecido cerebral de aves representando DVNP. No entanto, os níveis de atividade da AChE variaram de 27,6 a 79,9 µmol / min / g de tecido cerebral de aves representando áreas de cultivo. Os níveis de atividade de AChE observados no tagarela da selva, tagarela comum e bulbul vermelho exalado mostraram diferenças significativas (P 0,05). A inibição máxima foi registrada no tagarela da selva (53%), seguido pelo tagarela comum (35%), bulbul vermelho (18%), alvéola branca (15%) e drongo preto (7%). Os níveis de inibição da colinesterase cerebral nos ecossistemas subprotegidos (DVNP, Bhimber) e na paisagem agrícola sugerem contaminação por inseticida e seu impacto na diversidade da avifauna. O estudo também enfatiza a importância das zonas livres de pesticidas para proteger a biodiversidade das aves.

Assessment of Cholinesterase inhibition activity of birds inhabiting pesticide exposed croplands and protected area in hot semi-arid region of Pakistan / Avaliação da atividade de inibição da colinesterase de pássaros que habitam áreas cultivadas e áreas protegidas expostas a pesticidas na região semiárida quente do Paquistão

Acetylcholinesterase (AChE) activity levels can be used as an indicator for AChE inhibition due to pesticide poisoning in bird species. We assessed the comparative brain cholinesterase (AChE) activity level of five bird species inhabiting pesticide exposed croplands and Protected Area i.e. Deva Vatala National Park (DVNP), Bhimber by using a spectrophotometric method. AChE activity levels ranged from 56.3 to 85.9 µmol/min/g of brain tissue of birds representing DVNP. However, AChE activity levels ranged from 27.6 to 79.9 µmol/min/g of brain tissue of birds representing croplands. AChE activity levels observed in Jungle babbler, Common babbler, and Red-vented bulbul showed significant differences (P < 0.05) at two sites. However, White wagtail and Black drongo demonstrated non-significant differences (P > 0.05). Maximum inhibition was recorded in Jungle babbler (53%) followed by Common babbler (35%), Red-vented bulbul (18%), White wagtail (15%), and Black drongo (7%). The brain cholinesterase inhibition levels under-protected ecosystems (DVNP, Bhimber) and agricultural landscape suggest insecticidal contamination and its impact on avifauna diversity. The study also emphasizes on the importance of pesticide-free zones to protect the biodiversity of birds.(AU)

Os níveis de atividade da acetilcolinesterase (AChE) podem ser usados como um indicador para a inibição da AChE devido ao envenenamento por pesticidas em espécies de aves. Avaliamos o nível de atividade comparativa da colinesterase cerebral (AChE) de cinco espécies de aves que habitam áreas cultivadas expostas a pesticidas e Área Protegida, ou seja, Deva Vatala National Park (DVNP), Bhimber, usando um método espectrofotométrico. Os níveis de atividade da AChE variaram de 56,3 a 85,9 µmol / min / g de tecido cerebral de aves representando DVNP. No entanto, os níveis de atividade da AChE variaram de 27,6 a 79,9 µmol / min / g de tecido cerebral de aves representando áreas de cultivo. Os níveis de atividade de AChE observados no tagarela da selva, tagarela comum e bulbul vermelho exalado mostraram diferenças significativas (P < 0,05) em dois locais. No entanto, alvéola branca e drongo preto demonstraram diferenças não significativas (P > 0,05). A inibição máxima foi registrada no tagarela da selva (53%), seguido pelo tagarela comum (35%), bulbul vermelho (18%), alvéola branca (15%) e drongo preto (7%). Os níveis de inibição da colinesterase cerebral nos ecossistemas subprotegidos (DVNP, Bhimber) e na paisagem agrícola sugerem contaminação por inseticida e seu impacto na diversidade da avifauna. O estudo também enfatiza a importância das zonas livres de pesticidas para proteger a biodiversidade das aves.(AU)

Kinetic and toxicological effects of synthesized palladium(II) complex on snake venom (Bungarus sindanus) acetylcholinesterase

The venom of the krait (Bungarus sindanus), an Elapidae snake, is highly toxic to humans and contains a great amount of acetylcholinesterase (AChE). The enzyme AChE provokes the hydrolysis of substrate acetylcholine (ACh) in the nervous system and terminates nerve impulse. Different inhibitors inactivate AChE and lead to ACh accumulation and disrupted neurotransmission. Methods: The present study was designed to evaluate the effect of palladium(II) complex as antivenom against krait venom AChE using kinetics methods. Results: Statistical analysis showed that krait venom AChE inhibition decreases with the increase of Pd(II) complex (0.025-0.05 µM) and exerted 61% inhibition against the AChE at a fixed concentration (0.5 mM) of ACh. Kinetic analysis using the Lineweaver Burk plot showed that Pd(II) caused a competitive inhibition. The compound Pd(II) complex binds at the active site of the enzyme. It was observed that K m (Michaelis-Menten constant of AChE-ACh into AChE and product) increased from 0.108 to 0.310 mM (45.74 to 318.35%) and V max remained constant with an increase of Pd(II) complex concentrations. In AChE K Iapp was found to increase from 0.0912 to 0.025 µM (29.82-72.58%) and did not affect the V maxapp with an increase of ACh from (0.05-1 mM). K i (inhibitory constant) was estimated to be 0.029µM for snake venom; while the K m was estimated to be 0.4 mM. The calculated IC50 for Pd(II) complex was found to be 0.043 µM at constant ACh concentration (0.5 mM). Conclusions: The results show that the Pd(II) complex can be deliberated as an inhibitor of AChE.(AU)

Kinetic and toxicological effects of synthesized palladium(II) complex on snake venom (Bungarus sindanus) acetylcholinesterase

The venom of the krait (Bungarus sindanus), an Elapidae snake, is highly toxic to humans and contains a great amount of acetylcholinesterase (AChE). The enzyme AChE provokes the hydrolysis of substrate acetylcholine (ACh) in the nervous system and terminates nerve impulse. Different inhibitors inactivate AChE and lead to ACh accumulation and disrupted neurotransmission. Methods: The present study was designed to evaluate the effect of palladium(II) complex as antivenom against krait venom AChE using kinetics methods. Results: Statistical analysis showed that krait venom AChE inhibition decreases with the increase of Pd(II) complex (0.025-0.05 µM) and exerted 61% inhibition against the AChE at a fixed concentration (0.5 mM) of ACh. Kinetic analysis using the Lineweaver Burk plot showed that Pd(II) caused a competitive inhibition. The compound Pd(II) complex binds at the active site of the enzyme. It was observed that K m (Michaelis-Menten constant of AChE-ACh into AChE and product) increased from 0.108 to 0.310 mM (45.74 to 318.35%) and V max remained constant with an increase of Pd(II) complex concentrations. In AChE K Iapp was found to increase from 0.0912 to 0.025 µM (29.82-72.58%) and did not affect the V maxapp with an increase of ACh from (0.05-1 mM). K i (inhibitory constant) was estimated to be 0.029µM for snake venom; while the K m was estimated to be 0.4 mM. The calculated IC50 for Pd(II) complex was found to be 0.043 µM at constant ACh concentration (0.5 mM). Conclusions: The results show that the Pd(II) complex can be deliberated as an inhibitor of AChE.(AU)

Neuroactive venom compounds obtained from Phlogiellus bundokalbo as potential leads for neurodegenerative diseases: insights on their acetylcholinesterase and beta-secretase inhibitory activities in vitro

Spider venom is a rich cocktail of neuroactive compounds designed to prey capture and defense against predators that act on neuronal membrane proteins, in particular, acetylcholinesterases (AChE) that regulate synaptic transmission through acetylcholine (ACh) hydrolysis - an excitatory neurotransmitter - and beta-secretases (BACE) that primarily cleave amyloid precursor proteins (APP), which are, in turn, relevant in the structural integrity of neurons. The present study provides preliminary evidence on the therapeutic potential of Phlogiellus bundokalbo venom against neurodegenerative diseases. Methods Spider venom was extracted by electrostimulation and fractionated by reverse-phase high-performance liquid chromatography (RP-HPLC) and characterized by matrix-assisted laser desorption ionization-time flight mass spectrometry (MALDI-TOF-MS). Neuroactivity of the whole venom was observed by a neurobehavioral response from Terebrio molitor larvae in vivo and fractions were screened for their inhibitory activities against AChE and BACE in vitro. Results The whole venom from P. bundokalbo demonstrated neuroactivity by inducing excitatory movements from T. molitor for 15 min. Sixteen fractions collected produced diverse mass fragments from MALDI-TOF-MS ranging from 900-4500 Da. Eleven of sixteen fractions demonstrated AChE inhibitory activities with 14.34% (± 2.60e-4) to 62.05% (± 6.40e-5) compared with donepezil which has 86.34% (± 3.90e-5) inhibition (p > 0.05), while none of the fractions were observed to exhibit BACE inhibition. Furthermore, three potent fractions against AChE, F1, F3, and F16 displayed competitive and uncompetitive inhibitions compared to donepezil as the positive control. Conclusion The venom of P. bundokalbo contains compounds that demonstrate neuroactivity and anti-AChE activities in vitro, which could comprise possible therapeutic leads for the development of cholinergic compounds against neurological diseases.(AU)

Neuroactive venom compounds obtained from Phlogiellus bundokalbo as potential leads for neurodegenerative diseases: insights on their acetylcholinesterase and beta-secretase inhibitory activities in vitro

Spider venom is a rich cocktail of neuroactive compounds designed to prey capture and defense against predators that act on neuronal membrane proteins, in particular, acetylcholinesterases (AChE) that regulate synaptic transmission through acetylcholine (ACh) hydrolysis - an excitatory neurotransmitter - and beta-secretases (BACE) that primarily cleave amyloid precursor proteins (APP), which are, in turn, relevant in the structural integrity of neurons. The present study provides preliminary evidence on the therapeutic potential of Phlogiellus bundokalbo venom against neurodegenerative diseases. Methods Spider venom was extracted by electrostimulation and fractionated by reverse-phase high-performance liquid chromatography (RP-HPLC) and characterized by matrix-assisted laser desorption ionization-time flight mass spectrometry (MALDI-TOF-MS). Neuroactivity of the whole venom was observed by a neurobehavioral response from Terebrio molitor larvae in vivo and fractions were screened for their inhibitory activities against AChE and BACE in vitro. Results The whole venom from P. bundokalbo demonstrated neuroactivity by inducing excitatory movements from T. molitor for 15 min. Sixteen fractions collected produced diverse mass fragments from MALDI-TOF-MS ranging from 900-4500 Da. Eleven of sixteen fractions demonstrated AChE inhibitory activities with 14.34% (± 2.60e-4) to 62.05% (± 6.40e-5) compared with donepezil which has 86.34% (± 3.90e-5) inhibition (p > 0.05), while none of the fractions were observed to exhibit BACE inhibition. Furthermore, three potent fractions against AChE, F1, F3, and F16 displayed competitive and uncompetitive inhibitions compared to donepezil as the positive control. Conclusion The venom of P. bundokalbo contains compounds that demonstrate neuroactivity and anti-AChE activities in vitro, which could comprise possible therapeutic leads for the development of cholinergic compounds against neurological diseases.(AU)

Avaliação neurocomportamental e níveis de acetilcolinesterase cerebral em ratos expostos subcronicamente ao fipronil / Neurobehavioral assessment and brain acetylcholinesterase levels in rats subchronically exposed to fipronil

O fipronil é um inseticida de toxicidade seletiva amplamente empregado na agricultura e na medicina veterinária. Porém, há relatos de efeitos neurotóxicos dessa substância, que geram prejuízos para vertebrados. Avaliou-se a atividade locomotora, a coordenação motora e a atividade da enzima acetilcolinesterase cerebral em ratos expostos ao fipronil. Ratos Wistar machos adultos (n=15) receberam fipronil em dose de 30mg/kg, por via oral, durante 15 dias; o grupo controle (n=15) foi tratado com solução fisiológica, por via oral, no mesmo período. No 16° dia de experimentação, os animais foram submetidos aos testes de arena de campo aberto e hole board. No 17° dia, foram anestesiados e eutanasiados, procedendo-se à coleta de órgãos, e posteriormente foi feita a avaliação da AChE cerebral. A exposição ao fipronil não provocou alterações significativas sobre a coordenação motora e a atividade locomotora, porém gerou inibição significativa da atividade da acetilcolinesterase cerebral. Esses achados sugerem que o fipronil pode provocar efeitos neurotóxicos em curto prazo, os quais podem ser exacerbados caso a exposição seja prolongada.(AU)

Fipronil is a selective-toxicity insecticide widely used in agriculture and veterinary medicine. However, there are reports of neurotoxic effects of this substance, causing damages to vertebrates. We evaluated the locomotor activity, motor coordination and the activity of brain acetylcholinesterase in rats exposed to fipronil. Adult male Wistar rats (n= 15) received fipronil at a dose of 30mg/kg orally for 15 days; the Control group (n= 15) was treated with oral solution in the same period. On the 16th day of experimentation, the animals were submitted to the open field arena test and hole-board test. On the 17th day, they were anesthetized and euthanized, and organs were collected, and subsequently brain AChE was evaluated. Exposure to fipronil yielded no significant changes on motor coordination and locomotor activity but caused significant inhibition of brain acetylcholinesterase activity. These findings suggest that fipronil may cause short-term neurotoxic effects, which may be exacerbated if exposure is prolonged.(AU)

Avaliação neurocomportamental e níveis de acetilcolinesterase cerebral em ratos expostos subcronicamente ao fipronil / Neurobehavioral assessment and brain acetylcholinesterase levels in rats subchronically exposed to fipronil

O fipronil é um inseticida de toxicidade seletiva amplamente empregado na agricultura e na medicina veterinária. Porém, há relatos de efeitos neurotóxicos dessa substância, que geram prejuízos para vertebrados. Avaliou-se a atividade locomotora, a coordenação motora e a atividade da enzima acetilcolinesterase cerebral em ratos expostos ao fipronil. Ratos Wistar machos adultos (n=15) receberam fipronil em dose de 30mg/kg, por via oral, durante 15 dias; o grupo controle (n=15) foi tratado com solução fisiológica, por via oral, no mesmo período. No 16° dia de experimentação, os animais foram submetidos aos testes de arena de campo aberto e hole board. No 17° dia, foram anestesiados e eutanasiados, procedendo-se à coleta de órgãos, e posteriormente foi feita a avaliação da AChE cerebral. A exposição ao fipronil não provocou alterações significativas sobre a coordenação motora e a atividade locomotora, porém gerou inibição significativa da atividade da acetilcolinesterase cerebral. Esses achados sugerem que o fipronil pode provocar efeitos neurotóxicos em curto prazo, os quais podem ser exacerbados caso a exposição seja prolongada.(AU)

Fipronil is a selective-toxicity insecticide widely used in agriculture and veterinary medicine. However, there are reports of neurotoxic effects of this substance, causing damages to vertebrates. We evaluated the locomotor activity, motor coordination and the activity of brain acetylcholinesterase in rats exposed to fipronil. Adult male Wistar rats (n= 15) received fipronil at a dose of 30mg/kg orally for 15 days; the Control group (n= 15) was treated with oral solution in the same period. On the 16th day of experimentation, the animals were submitted to the open field arena test and hole-board test. On the 17th day, they were anesthetized and euthanized, and organs were collected, and subsequently brain AChE was evaluated. Exposure to fipronil yielded no significant changes on motor coordination and locomotor activity but caused significant inhibition of brain acetylcholinesterase activity. These findings suggest that fipronil may cause short-term neurotoxic effects, which may be exacerbated if exposure is prolonged.(AU)

Experimental infection by Brucella ovis: changes in NTPDase, 5’-nucleotidase and acetylcholinesterase associated cerebral oxidative stress

Background: Changes in purinergic and cholinergic signaling have been demonstrated in various pathologies associatedwith inflammation; however, the changes in brucellosis caused by the Gram-negative coccobacillus Brucella ovis are notknown. B. ovis is generally asymptomatic in ewes. Hepatosplenomegaly has been described in B. ovis, a non-zoonoticspecies, characterized by an extravascular inflammatory response. Purinergic system enzymes are closely involved withthe modulation of the immune system, pro- and anti-inflammatory events. The objective of this study was to investigatethe role of ectonucleotidases and cholinesterase’s in the brains of mice experimentally infected with B. ovis.Materials, Methods & Results: Forty-eight animals were divided into two groups: control (n = 24) and infected (n = 24).In group infected, 100 µL containing 1.3 x 107 UFC B. ovis /mL via intraperitoneal was used in inoculation. The brainswere collected from the animals on days 7, 15, 30 and 60 post-infection (PI). We measured levels of TBARS (substancesreactive to thiobarbituric acid) and ROS (reactive oxygen species) in the brain. The activity of NTPDase (using ATP andADP as substrate) and 5’-nucleotidase (using AMP as substrate) were evaluated in brain in addition to histopathologicalanalysis. No histopathological lesions were observed in the control group nor the infected group at days 7, 15, and 30 PI.However, multifocal areas with moderate microgliosis in the cerebral cortex were observed at day 60 PI in the infectedanimals. B. ovis DNA was detected in brain. During the course of infection, B. ovis caused greater lipid peroxidation inthe brains of infected animals than in the control group at day 60 PI. No significant results were observed at 7, 15 or day30 PI. Similarly, there was significantly more reactive oxygen species at day 60 PI in brains of infected animals than inthe control group. NTPDase activity (using ATP ...

Experimental infection by Brucella ovis: changes in NTPDase, 5-nucleotidase and acetylcholinesterase associated cerebral oxidative stress

Background: Changes in purinergic and cholinergic signaling have been demonstrated in various pathologies associatedwith inflammation; however, the changes in brucellosis caused by the Gram-negative coccobacillus Brucella ovis are notknown. B. ovis is generally asymptomatic in ewes. Hepatosplenomegaly has been described in B. ovis, a non-zoonoticspecies, characterized by an extravascular inflammatory response. Purinergic system enzymes are closely involved withthe modulation of the immune system, pro- and anti-inflammatory events. The objective of this study was to investigatethe role of ectonucleotidases and cholinesterases in the brains of mice experimentally infected with B. ovis.Materials, Methods & Results: Forty-eight animals were divided into two groups: control (n = 24) and infected (n = 24).In group infected, 100 µL containing 1.3 x 107 UFC B. ovis /mL via intraperitoneal was used in inoculation. The brainswere collected from the animals on days 7, 15, 30 and 60 post-infection (PI). We measured levels of TBARS (substancesreactive to thiobarbituric acid) and ROS (reactive oxygen species) in the brain. The activity of NTPDase (using ATP andADP as substrate) and 5-nucleotidase (using AMP as substrate) were evaluated in brain in addition to histopathologicalanalysis. No histopathological lesions were observed in the control group nor the infected group at days 7, 15, and 30 PI.However, multifocal areas with moderate microgliosis in the cerebral cortex were observed at day 60 PI in the infectedanimals. B. ovis DNA was detected in brain. During the course of infection, B. ovis caused greater lipid peroxidation inthe brains of infected animals than in the control group at day 60 PI. No significant results were observed at 7, 15 or day30 PI. Similarly, there was significantly more reactive oxygen species at day 60 PI in brains of infected animals than inthe control group. NTPDase activity (using ATP ...(AU)

Antimicrobial activity and acetylcholinesterase inhibition by extracts from chromatin modulated fungi

Major health challenges as the increasing number of cases of infections by antibiotic multiresistant microorganisms and cases of Alzheimer's disease have led to searching new control drugs. The present study aims to verify a new way of obtaining bioactive extracts from filamentous fungi with potential antimicrobial and acetylcholinesterase inhibitory activities, using epigenetic modulation to promote the expression of genes commonly silenced. For such finality, five filamentous fungal species (Talaromyces funiculosus, Talaromyces islandicus, Talaromyces minioluteus, Talaromyces pinophilus, Penicillium janthinellum) were grown or not with DNA methyltransferases inhibitors (procainamide or hydralazine) and/or a histone deacetylase inhibitor (suberohydroxamic acid). Extracts from T. islandicus cultured or not with hydralazine inhibited Listeria monocytogenes growth in 57.66 ± 5.98% and 15.38 ± 1.99%, respectively. Increment in inhibition of acetylcholinesterase activity was observed for the extract from P. janthinellum grown with procainamide (100%), when compared to the control extract (39.62 ± 3.76%). Similarly, inhibition of acetylcholinesterase activity increased from 20.91 ± 3.90% (control) to 92.20 ± 3.72% when the tested extract was obtained from T. pinophilus under a combination of suberohydroxamic acid and procainamide. Concluding, increases in antimicrobial activity and acetylcholinesterase inhibition were observed when fungal extracts in the presence of DNA methyltransferases and/or histone deacetylase modulators were tested.(AU)

Passion fruit and apple: from residues to antioxidant, antimicrobial and anti-Alzheimers potential / Maracujá e maçã: de resíduos ao potencial antioxidante, antimicrobiano e anti-Alzheimer

The waste in the fruit production chain, including the juice and pulp industries, produces large quantities of leftover husks, seeds and bagasse. This volume of waste generates huge environmental and economic impact. The objective of this research was to determine the potential of using residues from passion fruit (Passiflora edulis) and apple (Malus domestica) varieties in the production of functional flours. Passion fruit flour showed greater reduction of DPPH (EC50%: 50.4μg/mL) radicals, showing antioxidant potential, as well as a more efficient inhibition of Staphylococcus aureus (71.3±1.2μg/mL), with a modest; however efficient, inhibition of acetylcholinesterase (10%). All Apple flours were good antioxidants and the fuji apple flour stood out inhibiting Pseudomonas aeruginosa (78.6±3.1μg/mL). All the residues showed potential for use as a functional product either as a source of antioxidants, a natural (antimicrobial) preservative for dry foods or supplementary use by patients with Alzheimers disease.(AU)

O desperdício na cadeia produtiva de frutos, incluindo as indústrias de suco e polpa, produz grandes quantidades de resíduos, como cascas, sementes e bagaços. Esse volume de resíduos gera impacto ambiental e econômico. O objetivo deste trabalho foi determinar o potencial de aproveitamento de resíduos de maracujá (Passiflora edulis) e de variedades de maçã (Malus domestica) na produção de farinhas funcionais. A farinha de maracujá apresentou maior redução de radicais DPPH (EC50%: 50,4μg/mL), demonstrando potencial antioxidante, maior inibição de Staphylococcus aureus (71,3±1,2μg/mL) e da acetilcolinesterase (10%). Todas as farinhas de maçã foram boas antioxidantes e a fuji destacou-se inibindo Pseudomonas aeruginosa (78,6±3,1μg/mL). Todos os resíduos mostraram potencial para aproveitamento como produto funcional, seja como fonte de antioxidantes, conservante natural (antimicrobiano) para alimentos secos ou uso suplementar no tratamento de Alzheimer.(AU)

Effect of Sciatic Nerve Transection on acetylcholinesterase activity in spinal cord and skeletal muscles of the bullfrog Lithobates catesbeianus / Efeito da Secção do Nervo Isquiático sobre a atividade da acetilcolinesterase em medula espinal e músculos esqueléticos da rã-touro Lithobates catesbeianus

Sciatic nerve transection (SNT), a model for studying neuropathic pain, mimics the clinical symptoms of "phantom limb", a pain condition that arises in humans after amputation or transverse spinal lesions. In some vertebrate tissues, this condition decreases acetylcholinesterase (AChE) activity, the enzyme responsible for fast hydrolysis of released acetylcholine in cholinergic synapses. In spinal cord of frog Rana pipiens, this enzymes activity was not significantly changed in the first days following ventral root transection, another model for studying neuropathic pain. An answerable question is whether SNT decreases AChE activity in spinal cord of frog Lithobates catesbeianus, a species that has been used as a model for studying SNT-induced neuropathic pain. Since each animal model has been created with a specific methodology, and the findings tend to vary widely with slight changes in the method used to induce pain, our study assessed AChE activity 3 and 10 days after complete SNT in lumbosacral spinal cord of adult male bullfrog Lithobates catesbeianus. Because there are time scale differences of motor endplate maturation in rat skeletal muscles, our study also measured the AChE activity in bullfrog tibial posticus (a postural muscle) and gastrocnemius (a typical skeletal muscle that is frequently used to study the motor system) muscles. AChE activity did not show significant changes 3 and 10 days following SNT in spinal cord. Also, no significant change occurred in AChE activity in tibial posticus and gastrocnemius muscles at day 3. However, a significant decrease was found at day 10, with reductions of 18% and 20% in tibial posticus and gastrocnemius, respectively. At present we cannot explain this change in AChE activity. While temporally different, the direction of the change was similar to that described for rats.[...](AU)

A transecção do nervo isquiático (SNT), um modelo para estudar dor neuropática, simula os sintomas clínicos do membro fantasma, uma condição dolorosa que ocorre nos humanos após amputação ou secção completa da medula espinal. Essa condição muda a atividade da acetilcolinesterase (AChE), a enzima responsável pela rápida hidrólise da acetilcolina liberada nas sinapses colinérgicas, em alguns tecidos de vertebrados. Em medula espinal de rã Rana pipiens, a atividade da AChE não foi significativamente alterada nos primeiros dias após a secção da raiz ventral, outro modelo para o estudo da dor neuroptípica. Uma questão ainda não respondida é se a SNT diminui a atividade da AChE na medula espinal de rã Lithobates catesbeianus, uma espécie que vem sendo usada como modelo em estudos da dor neuropática induzida por SNT. Como cada modelo animal é criado a partir de metodologia específica, e seus resultados tendem a variar com pequenas mudanças na metodologia de indução da dor, o presente estudo avaliou a atividade da AChE em medula espinal lombossacral de rã-touro Lithobates catesbeianus, adultos, machos, 3 e 10 dias após a completa SNT. Como há diferenças temporais na maturação de placas motoras em músculos esqueléticos de ratos, nosso estudo ainda demonstrou, na rã-touro, os efeitos da SNT sobre a atividade da AChE nos músculos esqueléticos tibial posticus, um músculo postural, e gastrocnêmio, um músculo frequentemente usado em estudos do sistema motor. A atividade da AChE não mudou significativamente na medula espinal aos 3 e 10 dias após a SNT. Nos músculos, a atividade não alterou significativamente aos 3 dias após a lesão, mas reduziu de forma significativa aos 10 dias após a SNT. Aos 10 dias, a diminuição foi 18% no músculo tibial posticus e 20% no gastrocnêmio. No momento, nós não temos explicação para essa mudança na atividade da AChE. Embora temporalmente diferente, o sentido da mudança é similar ao que é descrito em ratos.[...](AU)

Biomarcadores inflamatórios e de estresse oxidativo em cadelas submetidas à ovário-histerectomia videoassistida ou convencional / Inflammation and oxidative stress indicators in bitches submitted to laparoscopic-assisted or conventional ovariohysterectomy

A videocirurgia é atualmente uma das principais ferramentas operatórias, com vantagens que incluem menor estresse, incisões e dor pós-operatória quando comparada aos procedimentos abertos. Objetivou-se comparar o processo inflamatório e o estresse oxidativo resultantes das técnicas de ovário-histerectomia (OVH) convencional e videoassistida, com dois portais em cadelas, por meio de hemograma, avaliação de acetilcolinesterase, butirilcolinesterase, catalase e malondialdeído séricos, imediatamente antes da operação e duas, seis, 12, 24, 48 e 72 horas após a cirurgia. Observou-se menor estresse cirúrgico nas pacientes operadas pela técnica videoassistida, e sugere-se que a técnica convencional possa implicar peroxidação lipídica, mesmo com o uso de anti-inflamatório.(AU)

Videosurgery is currently a very important surgical tool with several benefits over the open surgery, including less surgical stress, shorter incisions and less postoperative pain. The purpose of this study was to compare the inflammatory process and oxidative stress between conventional and two-port laparoscopic-assisted ovariohisterectomy (OVH) in bitches. Complete blood counting, serum acetylcholinesterase, butyrylcholinesterase, catalase and malondialdehyde were assessed on the baseline and at two, six, 12, 24, 48 and 72 hours after surgery. The patients submitted to the videoassisted technique presented lower inflammatory response. There are suggestions that the conventional technique promotes lipid peroxidation, even with the use of anti-inflammatories.(AU)

Biomarcadores inflamatórios e de estresse oxidativo em cadelas submetidas à ovário-histerectomia videoassistida ou convencional / Inflammation and oxidative stress indicators in bitches submitted to laparoscopic-assisted or conventional ovariohysterectomy

A videocirurgia é atualmente uma das principais ferramentas operatórias, com vantagens que incluem menor estresse, incisões e dor pós-operatória quando comparada aos procedimentos abertos. Objetivou-se comparar o processo inflamatório e o estresse oxidativo resultantes das técnicas de ovário-histerectomia (OVH) convencional e videoassistida, com dois portais em cadelas, por meio de hemograma, avaliação de acetilcolinesterase, butirilcolinesterase, catalase e malondialdeído séricos, imediatamente antes da operação e duas, seis, 12, 24, 48 e 72 horas após a cirurgia. Observou-se menor estresse cirúrgico nas pacientes operadas pela técnica videoassistida, e sugere-se que a técnica convencional possa implicar peroxidação lipídica, mesmo com o uso de anti-inflamatório.(AU)

Videosurgery is currently a very important surgical tool with several benefits over the open surgery, including less surgical stress, shorter incisions and less postoperative pain. The purpose of this study was to compare the inflammatory process and oxidative stress between conventional and two-port laparoscopic-assisted ovariohisterectomy (OVH) in bitches. Complete blood counting, serum acetylcholinesterase, butyrylcholinesterase, catalase and malondialdehyde were assessed on the baseline and at two, six, 12, 24, 48 and 72 hours after surgery. The patients submitted to the videoassisted technique presented lower inflammatory response. There are suggestions that the conventional technique promotes lipid peroxidation, even with the use of anti-inflammatories.(AU)

In vitro screening and evaluation of antivenom phytochemicals from Azima tetracantha Lam. leaves against Bungarus caeruleus and Vipera russelli

Snakebites are considered a neglected tropical disease that affects thousands of people worldwide. Although antivenom is the only treatment available, it is associated with several side effects. As an alternative, plants have been extensively studied in order to obtain an alternative treatment. In folk medicine, Azima tetracantha Lam. is usually used to treat snakebites. The present study aims to provide a scientific explanation for the use of this plant against snakebite. The extracts of shade dried leaves of A. tetracantha were tested for in vitro inhibitory activity on toxic venom enzymes like phosphomonoesterase, phosphodiesterase, acetylcholinesterase, hyaluronidase etc. from Bungarus caeruleus and Vipera russelli venoms.(AU)

In vitro screening and evaluation of antivenom phytochemicals from Azima tetracantha Lam. leaves against Bungarus caeruleus and Vipera russelli

Snakebites are considered a neglected tropical disease that affects thousands of people worldwide. Although antivenom is the only treatment available, it is associated with several side effects. As an alternative, plants have been extensively studied in order to obtain an alternative treatment. In folk medicine, Azima tetracantha Lam. is usually used to treat snakebites. The present study aims to provide a scientific explanation for the use of this plant against snakebite. The extracts of shade dried leaves of A. tetracantha were tested for in vitro inhibitory activity on toxic venom enzymes like phosphomonoesterase, phosphodiesterase, acetylcholinesterase, hyaluronidase etc. from Bungarus caeruleus and Vipera russelli venoms.

Changes in oxidative markers, endogenous antioxidants and activity of the enzyme acetylcholinesterase in farmers exposed to agricultural pesticides: a pilot study / Alterações em marcadores oxidativos, antioxidantes endógenos e na atividade da enzima acetilcolinesterase em trabalhadores rurais expostos a pesticidas agrícolas: um estudo piloto

Agricultural practices are directly related to the use of pesticides, which indiscriminately and without due care may contribute to the occurrence of numerous intoxications. Several studies have demonstrated the relationship of certain pesticides and the occurrence of oxidative stress and therefore, in recent years have developed methods of analysis of several biomarkers of cellular damage that can be measured and quantified. In this context, the objective of this study was to evaluate the possible changes in biochemical markers: glutamic aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, urea, total protein, and oxidative markers such as lipid peroxidation, damage to proteins and the activity of the enzymes acetylcholinesterase (AChE) and catalase (CAT) in farmers exposed to different pesticides for at least five years from Ibirubá - RS city. With the exception of AST and ALT, the results showed a significant difference between the mean total protein, urea and creatinine in the control group, showing that no changes in liver or kidney function of rural workers. In the oxidative parameters, there was a decrease in AChE activity and CAT in the control group; there were an increase in protein carbonyl and a decreased on TBARS compared to control group. Therefore, the results demonstrated a change in oxidative status of rural workers compared with the control group, mainly by possible inhibition of AChE activity and the occurrence of oxidative stress without showing changes in biochemical parameters.(AU)

As práticas agrícolas estão diretamente relacionadas com o uso de agrotóxico, que, de forma indiscriminada e sem o devido cuidado, podem contribuir para a ocorrência de inúmeras intoxicações. Diversos estudos demonstraram a relação de determinados pesticidas e a ocorrência do estresse oxidativo e, portanto, nos anos recentes, tem-se desenvolvido métodos de análise de diversos biomarcadores de dano celular, o qual pode ser medido e quantificado. Nesse contexto, o objetivo deste estudo foi avaliar as possíveis alterações nos marcadores bioquímicos: aspartato aminotransferase (AST), alanina aminotransferase (ALT), creatinina, ureia, proteína total, e marcadores oxidativos como: a peroxidação de lipídios, danos nas proteínas e a atividade das enzimas acetilcolinesterase (AChE) e catalase (CAT), em agricultores da cidade de Ibirubá, RS, expostos a diferentes agrotóxicos por pelo menos cinco anos. Com exceção da AST e da ALT, os resultados mostraram diferença significativa entre as médias das proteínas totais, ureia e creatinina, no grupo controle, mostrando que não houve alterações na função hepática ou renal dos trabalhadores rurais. Nos parâmetros oxidativos avaliados, houve uma diminuição da atividade da AChE e da CAT em relação ao grupo controle; um aumento das proteínas carboniladas e uma diminuição dos níveis de TBARS em relação ao grupo controle. Portanto, os resultados demonstram uma alteração oxidativa nos trabalhadores rurais, comparados com o grupo controle, principalmente pela possível inibição da AChE e ocorrência do estresse oxidativo, sem demonstrar mudança nos parâmetros bioquímicos analisados.(AU)

Enzymatic and biochemical characterization of Bungarus sindanus snake venom acetylcholinesterase

This study analyses venom from the elapid krait snake Bungarus sindanus, which contains a high level of acetylcholinesterase (AChE) activity. The enzyme showed optimum activity at alkaline pH (8.5) and 45ºC. Krait venom AChE was inhibited by substrate. Inhibition was significantly reduced by using a high ionic strength buffer; low ionic strength buffer (10 mM PO4 pH 7.5) inhibited the enzyme by 1. 5mM AcSCh, while high ionic strength buffer (62 mM PO4 pH 7.5) inhibited it by 1 mM AcSCh. Venom acetylcholinesterase was also found to be thermally stable at 45ºC; it only lost 5% of its activity after incubation at 45ºC for 40 minutes. The Michaelis-Menten constant (Km) for acetylthiocholine iodide hydrolysis was found to be 0.068 mM. Krait venom acetylcholinesterase was also inhibited by ZnCl2, CdCl2, and HgCl2 in a concentrationdependent manner. Due to the elevated levels of AChE with high catalytic activity and because it is more stable than any other sources, Bungarus sindanus venom is highly valuable for biochemical studies of this enzyme.(AU)