Anthelmintic activity of nanoencapsulated carvacryl acetate against gastrointestinal nematodes of sheep and its toxicity in rodents / Atividade anti-helmíntica do acetato de carvacrila nanoencapsulado sobre nematoides gastrintestinais de ovinos e toxicidade em roedores

Abstract The objective of this study was to evaluate the efficacy of carvacryl acetate (CVA) and nanoencapsulated CVA (nCVA) on gastrointestinal nematodes of sheep. The CVA was nanoencapsulated with chitosan/gum arabic and the efficacy of nanoencapsulation (EE), yield, zeta potential, nanoparticle morphology and release kinetics at pH 3 and 8 were analyzed. Acute and subchronic toxicity were evaluated in rodents and reduction of egg counts in the faeces (FECRT) of sheep. The sheep were divided into four groups (n = 10): G1, 250 mg/kg CVA; G2, 250 mg/kg nCVA; G3, polymer matrix and G4: 2.5 mg/kg monepantel. EE and nCVA yield were 65% and 57%, respectively. The morphology of the nanoparticles was spherical, size (810.6±286.7 nm), zeta potential in pH 3.2 (+18.3 mV) and the 50% release of CVA at pHs 3 and 8 occurred at 200 and 10 h, respectively. nCVA showed LD50 of 2,609 mg/kg. CVA, nCVA and monepantel reduced the number of eggs per gram of faeces (epg) by 57.7%, 51.1% and 97.7%, respectively. The epg of sheep treated with CVA and nCVA did not differ from the negative control (P>0.05). Nanoencapsulation reduced the toxicity of CVA; however, nCVA and CVA presented similar results in the FECRT.

Resumo O objetivo deste trabalho foi avaliar a eficácia do acetato de carvacrila (ACV) e do ACV nanoencapsulado (nACV) sobre nematóides gastrintestinais de ovinos. O ACV foi nanoencapsulado com quitosana/goma arábica e foi analisada a eficácia de nanoencapsulamento (EE), o rendimento, potencial zeta, morfologia das nanopartículas e cinética de liberação em pH 3 e 8. Foram avaliadas as toxicidades aguda e subcrônica em roedores e a redução da contagem de ovos nas fezes (RCOF) de ovinos. Os ovinos foram divididos em quatro grupos (n = 10): G1, 250 mg/kg ACV; G2, 250 mg/kg de nACV; G3, matriz polimérica e G4: 2,5 mg/kg de monepantel. A EE e o rendimento de nACV foram de 65% e 57%, respectivamente. A morfologia das nanopartículas foi esférica, tamanho (810,6±286,7 nm), potencial zeta no pH 3,2 (+18,3 mV) e a liberação de 50% de CVA nos pHs 3 e 8 ocorreu às 200 e 10 h, respectivamente. nACV apresentou DL50 de 2.609 mg/kg. ACV, nACV e o monepantel reduziram a contagem de ovos por grama de fezes (opg) em 57,7%, 51,1% e 97,7%, respectivamente. A contagem de opg de ovelhas tratadas com ACV e nCVA não diferiu do controle negativo (P>0,05). O nanoencapsulamento reduziu a toxicidade do AVC; no entanto, nACV e ACV apresentaram resultados semelhantes na RCOF.

Chronic treatment with carvacrol improves passive avoidance memory in a rat model of Parkinson's disease / O tratamento com carvacrol melhora a memória de esquiva passiva em um modelo da doença de Parkinson em ratos

ABSTRACT The present study investigated the effects of carvacrol on motor and memory deficits as well as hyperalgesia in the 6-OHDA-lesioned rat model of Parkinson's disease. The animals were subjected to unilateral microinjection of 6-OHDA into the medial forebrain bundle and treated with carvacrol (25, 50 and 100 mg/kg, ip) for six weeks after surgery. The 6-OHDA-lesioned rats showed contralateral rotations towards the lesion side, which was accompanied by learning and memory deficits in a passive avoidance test and a decrease in tail withdrawal latency in a tail flick test at the end of week 6. The results also showed that treatment with carvacrol at a dose of 25 mg/kg ameliorated memory deficits, with no effect on rotations and hyperalgesia in lesioned rats. In conclusion, carvacrol improves memory impairments in rats with Parkinson's disease; therefore, it may serve as an adjunct therapy for the alleviation of memory deficits in Parkinson's disease patients.

RESUMO O presente estudo investigou os efeitos do carvacrol nos déficits motores e de memória, bem como na hiperalgesia, em um modelo da doença de Parkinson (DP) em ratos com lesões 6-OHDA. Os animais foram submetidos a microinjeção unilateral de 6-OHDA no feixe mediano do prosencéfalo e tratados com carvacrol (25, 50 e 100 mg / kg, ip) durante 6 semanas após a cirurgia. Os ratos com lesões 6-OHDA mostraram rotações contralaterais para o lado da lesão, que foram acompanhadas de déficits de aprendizagem e de memória em um teste de evitação passiva, e de uma diminuição da latência de retirada da cauda em um teste de cauda no final da semana 6. Os resultados também mostraram que o tratamento crônico com carvacrol a uma dose de 25 mg / kg aliviou os déficits de memória, sem efeito sobre rotações e hiperalgesia em ratos lesados. Em conclusão, o carvacrol melhora a deficiência de memória em ratos com DP e, portanto, pode servir como uma terapia complementar para aliviar os déficits de memória em pacientes com DP.

Antifungal activity of geraniol and citronellol against food-relevant dematiaceous fungi Cladosporium spp / Atividade antifúngica de geraniol e citronelol frente a fungos dematiáceos de importância para os alimentos do gênero Cladosporium spp

Cladosporium spp. is a group of dematiaceous food-relevant fungi which are well dispersed in the environment causing food spoilage and poisoning. Considering the importance of fungal contamination, natural drugs to control their growth have become important. Thus, the aim of this study was to evaluate the inhibitory effects of two monoterpenoids, (geraniol and citronellol), against strains of Cladosporium carrioni, C. cladosporioides,and C. oxysporum. Methods: The Minimum Inhibitory Concentration (MIC) and Minimum Fungicide Concentration (MFC) of the drugs were determined by microdilution. The effects of test drugs on mycelial dry weight, conidia germination, and conidiogenesis of Cladosporium spp. were also investigated using a hemacytometer. Respective MIC and MFC values of citronellol varied from 256 to 512 μg/mL, and from 256 to 2048 μg/mL. The MIC and MFC of geraniol varied similarly to citronellol. Conidia germination, mycelial dry weight, and conidiogenesis of Cladosporium spp. were reduced by the test-drugs at 1/2MIC, MIC and 2xMIC (p<0.05). These measurable cell events are essential for fungal infection and development in foods. The action of citronellol and geraniol against Cladosporium spp. suggest that the drugs may serve as effective agents for controlling fungal contamination and growth in foods.

Cladosporium spp. é um grupo de fungos dematiáceos relevantes para os alimentos, que podem ser dispersos pelo ambiente e causar deterioração e intoxicação alimentar. Considerando a importância da contaminação fúngica, os produtos naturais usados para controlar seu crescimento são importantes. neste contexto, o objetivo deste estudo foi avaliar os efeitos inibitórios de dois monoterpenoides, geraniol e citronelol, contra cepas de Cladosporium carrioni, C. cladosporioides e C. oxysporum. A Concentração Inibitória Mínima (CIM) e Concentração Fungicida Mínima (CFM) das drogas foram determinadas por microdiluição. os efeitos das drogas-teste sobre a massa micelial seca, a germinação de conídios e a conidiogênese de Cladosporium spp. também foram investigados utilizando um hemocitômetro. os valores de CIM e CFM do citronelol variaram de 256 a 512 μg/mL e de 256 a 2048 μg/mL, respectivamente. CIM e CFM de geraniol variaram de forma semelhante. A germinação de conídios, massa micelial seca e conidiogênese de Cladosporium spp. foram inibidas pelas drogas-teste 1/2CIM, CIM e 2xCIM (p<0,05). Esses eventos celulares são essenciais para a infecção e desenvolvimento fúngico em alimentos. A ação de citronelol e geraniol contra Cladosporium spp. sugere que podem servir como agentes eficazes para controlar a contaminação fúngica e seu crescimento em alimentos.

Característica clínica e topográfica do glioma maligno em adolescente - Influência do tratamento intranasal com álcool perílico / Clinical and topographic characteristics of primary glioma in youth - Influence the response to perillyl alcohol treatment

Objetivo: Analisar a influência da topografia da lesão tumoral na resposta ao tratamento intranasal com álcool perílico (POH) em jovens com glioma maligno recidivo. Método: Tendo como padrão a faixa etária de 0 a 19 anos, foram incluídos pacientes do sexo masculino (#153; #31) e feminino (#178) com glioma maligno em estágio terminal, recebendo terapia de suporte paliativa e administração intranasal diária de 440 mg de POH. Resultados: Cefaleia intensa, tontura, vômito, crise convulsiva, alteração de comportamento, fraqueza muscular, alteração visual e hemiplegia à direita foram os sintomas prevalentes antes da confirmação diagnóstica de glioma. Análise de imagem mostrou lesão tumoral nas regiões troncocerebral (#153), talamomesencefálica esquerda (#178) e frontotemporal e insular direita (#31). Paciente #178 não respondeu ao tratamento, evoluindo a óbito em três semanas, e paciente #31 permaneceu em tratamento com POH por aproximadamente 54 semanas. Apesar de nova recidiva, paciente #153 apresenta doença estável, sem qualquer evidência clínica de recorrência para mais de 200 semanas em tratamento exclusivo com álcool perílico por via intranasal. Conclusão: Pacientes adolescentes com glioma maligno recidivo apresentaram heterogeneidade de sintomas compatível coma região anatômica comprometida, indicando que a topografia da lesão tumoral foi um fator prognóstico de sobrevida, influenciando inclusive na resposta ao tratamento intranasal com o álcool perílico.

Objective: Analyze the influence of tumor topography on response to intranasal perillyl (POH) treatment in youths with high grade glioma. Method: It was included male patients (#153; #31) with 19 years old and female (#178) with 15 years old with recurrent high grade glioma in terminal stage using supportive therapy and 440 mg POH daily intranasal administration. It was established a relation of clinical data and topographic image with therapeutic response to intranasal POH. Results: Intense headache, dizziness, vomiting, seizures, behavior change, muscle weakness, visual changes and right hemiplegia were the symptoms prevalent before the diagnostic confirmation of glioma. Image analysis showed tumoral lesionin the brain-stem (#153), in the left thalamus-mesencephalic region (#178), and right frontal-temporal and insular regions (#31). Patient #178 did not respond to intranasal POH treatment and rapidly progressed to death within 3 weeks; patient #31 remained in treatment with POH for nearly 54 weeks, and despite new recurrence, patient #153 presents stable disease, without any clinical evidence of recurrence for more than 200 weeks and under treatment exclusively with POH by intranasal route. Conclusion: Childhood patients with high grade malignant glioma had heterogeneity of clinical symptoms compatible with anatomical compromised region indicating that topography of the tumoral lesion was a prognostic factor influencing the overall survival and response to intranasal POH.

Anti-inflammatory and redox-protective activities of citronellal

The anti-inflammatory and redox protective effects of the citronellal (CT) were evaluated using in vivo and in vitro tests. Intraperitoneal (i.p.) administration of CT (50, 100, and 200 mg/kg) inhibited (p < 0.05) the carrageenan-induced leukocyte migration to the peritoneal cavity. Additionally, the carrageenan- and arachidonic acid-induced rat hind paw edema was significantly inhibited (p < 0.05) by i.p. administration of 100 and 200 mg/kg of the compound. When the redox activity was evaluated, CT (200 mg/kg) significantly reduced hepatic lipoperoxidation (p < 0.001), as well as oxidation of plasmatic (p < 0.05) and hepatic (p < 0.01) proteins. The results of the present study support the hypothesis that CT possesses anti-inflammatory and redox protective activities. It is suggested that its effects are associated with the inhibition of the enzymes in the arachidonic acid pathway, which prevent cell migration by inhibiting leukotriene production, edema formation and the increase of reactive oxygen species in tissues. Therefore, CT is of potential benefit to manage inflammatory disorders and correlated damages caused by oxidant agents.

Correlação da topografia tumoral e edema peritumoral com a resposta terapêutica à administração intranasal do álcool perílico nos gliomas malignos recidivos / Correlation of tumor topography and peritumoral edema with therapeutic response to a new approach for recurrent malignant gliomas, intranasal administration of perillyl alcohol

Objetivo: Avaliar a correlação da topografia tumoral e edema peritumoral com a resposta terapêutica à administração intranasal do álcool perílico (AP) em uma coorte de pacientes com gliomas malignos recidivos. Métodos: Os autores revisaram retrospectivamente 67 pacientes com gliomas malignos recidivos que receberam administração intranasal de 440 mg de AP diariamente. Parâmetros avaliados incluíram aspectos clínicos e os de neuroimagem. Avaliação clínica incluiu dados demográficos, sintomas iniciais e sobrevida global. Dados de imagem incluíram topografia tumoral, volume tumoral, presença de desvio da linha média eextensão de edema peritumoral. Análise bioestatística foi realizada usando testes log rank. Sobrevida global foi medida e analisada pelo método de Kaplan Meier, incluindo intervalos de confiança de 95 por cento. Resultados: Um total de 67 pacientes foi investigado, 52 (77,6 por cento) com glioblastoma (GBM), 10 (14,9 por cento) com astrocitoma anaplásico (AA) e 5 (7,4 por cento) com oligodendrioglioma anaplásico (OA). Todos os cinco pacientes com AO apresentaram tumor de localização lobar. Nos AA, oito casos estavam localizados em região talâmica e dois em região lobar, enquanto que, nos GBM, 11 casos de localização talâmica e 41 lobares. A relação volume tumoral e edema peritumoral foi observada. Pacientes com regressão tumoral e edema peritumoral apresentaram resposta positiva enquanto que aqueles com regressão tumoral sem regressão do edema peritumoral não apresentaram boa evolução clínica. Pacientes com gliomas profundos sobreviveram significativamente mais tempo do que aqueles com gliomas lobares (log rank test, p = 0,0003). Presença de desvio da linha média (> 1 cm) foi estatisticamente significante como fator de risco para a sobrevida mais curta (log rank test,p = 0,0062). Conclusões...

To evaluate the correlation of tumor topography and peritumoral brain edema with the therapeutic response of intranasal administration of perillyl alcohol (POH) in a cohort of patients with recurrent malignant gliomas. Methods: We retrospectively reviewed 67 consecutive patients with recurrent malignant gliomas who received administration intranasal of POH 440 mg daily. The parameters evaluated were clinical features and the neuroimaging findings. Clinical data included demographics, initial symptoms, and overall survival.Imaging analysis included tumor topography, tumor size, presence of midline shift and extent of peritumoraledema. Biostatistics was carried out using log rank tests. Overall survival was measure and analyzedby Kaplan Meier method including 95% confidence intervals. A total of 67 patients were investigated,52 (77.6%) with glioblastoma (GBM), 10 (14.9%) with anaplastic astrocytoma (AA) and 5 (7.4%) withanaplastic oligodendroglioma (AO). All five AO had lobar localization, AA were lobar in 8 cases and deep in 2 cases, whereas GBM were lobar in 41 cases and deep in 11 cases. Results: A relationship between the tumor size and the volume of peritumoral brain edema (PTBE) was observed. Patients with regression of the tumor and PTBE had positive response whereas those with regression of the tumor without PTBE regression had poor prognosis. Patients with deep tumors survived significantly longer than those with lobar gliomas (log rank test, p=0.0003). Presence of midline shift (> 1 cm) was a statistically significant risk factor for shorter survival (log rank test, p=0.0062). Conclusions...

Antinociceptive effects of the essential oil of Mentha x villosa leaf and its major constituent piperitenone oxide in mice

Mentha x villosa Huds (Labiatae) is an aromatic herb widely used in folk medicine. Since the essential oil of the herb has many pharmacological activities, including antispasmodic effects, we determined whether the oil and its major constituent, piperitenone oxide (PO), have antinociceptive activity. The essential oil of M. x villosa (EOMV) and PO administered orally at 200 mg/kg (vehicle: 0.1% Tween 80 in water) significantly reduced the writhings induced by acetic acid from control values of 59.5 ± 3.1 s (N = 10) to 31.9 ± 2.8 s (N = 10) and 23.8 ± 3.4 s (N = 10), respectively. When administered at 100 and 200 mg/kg, EOMV reduced the paw licking time for the second phase of the formalin test from the control value of 20.6 ± 2.1 s (N = 13) to 5.3 ± 2.2 s (N = 12) and 2.7 ± 1.2 s (N = 18), respectively. At 100 and 200 mg/kg, PO reduced this second phase to 8.3 ± 2.7 s (N = 12) and 3.0 ± 1.2 s (N = 10), respectively. This effect of EOMV and PO was not reversed by naloxone. EOMV and PO had no significant effect on the first phase of the formalin test. As evaluated by the hot-plate and tail immersion test, EOMV and PO, at doses up to 200 mg/kg, showed no analgesic activity. These results show that EOMV and PO have antinociceptive activity and suggest that this effect is probably an indirect anti-inflammatory effect, which does not involve the central nervous system.

Comparative sem evaluation by of three solvents used in endodontic retreatment: an ex vivo study

This study compared, by scanning electron microscopy (SEM), the efficacy of three solvents on the removal of filling materials from dentinal tubules during endodontic retreatment. Forty human maxillary canines with straight canals were prepared according to a crown-down technique and enlarged to a#30 apical file size, before obturation with gutta-percha and a zinc-oxide-eugenol based sealer. The samples were stored for 3 months before being randomly assigned to four groups: chloroform (n=10), orange oil (n=10), eucalyptol (n=10) and control (n=10). Solvents were applied to a reservoir created on the coronal root third using Gates Glidden drills. The total time for retreatment using the solvents was 5 minutes per tooth. Following retreatment the roots were split longitudinally for SEM evaluation. SEM images were digitized, analyzed using Image ProPlus 4.5 software, and the number of dentinal tubules free of filling material from the middle and apical thirds was recorded. No significant difference was found among the solvent groups regarding the number of dentinal tubules free of root filling remnants in the middle and apical root thirds (p>0.05). However, the control group had fewer dentinal tubules free of filling material (p<0.05). Under the tested conditions, it may be concluded that there was no significant difference among the solvents used to obtain dentinal tubules free of filling material remnants.

Detecção imuno-histoquímica de células apoptóticas em glioblastoma recidivante de paciente tratado com álcool perílico por via inalatória / Immunohistochemical detection of apoptotic cells in a recurrent glioblastoma patient treated with intranasal delivery of perillyl alcohol

Introdução: Estudos in vitro mostram que radioterapia e/ou quimioterapia podem ativar as vias de sinalizaçãodo receptor do fator de crescimento epidérmico (EGFR) e Ras, aumentando a resistência cruzada dascélulas de glioblastomas multiformes (GBM) ao tratamento. A inibição das atividades de EGFR e Rasatravés de inibidores tirosinas cinases elimina o antagonismo observado à administração seqüencialdestas modalidades terapêuticas, induzindo apoptose nestas células. Em estudo prévio demonstramosque o tratamento com o álcool perílico (AP), inibidor da farnesilação da Ras, induz apoptose em linhagenscelulares e células de explante de GBM. Objetivo: No presente estudo investigamos se a regressãoparcial observada em GBM recorrente de paciente tratado com administração intranasal de AP é mediadapor apoptose. Resultado: Ensaios com TUNEL (deoxynucleotidyl-mediated deoxyuridine triphosphate) ecaspase-3 ativada evidenciaram presença de células apoptóticas nas lâminas de GBM tratado. Conclusão:Esses achados sugerem que estratégias adjuvantes visando à inativação das vias de sinalização do EGFRe Ras podem melhorar tanto a eficácia de terapia isolada como de terapia multimodal em gliomas.

Background: In vitro studies demonstrated that both radiation and chemotherapy can activate EGFRand Ras signaling pathways, leading to increased cross-resistance to treatment of GBM cell. Inhibition of either EGFR or Ras activity with tytosine kinase inhibitor appears to abrogate the observed antagonism between sequentially administration of these therapeutic modalities inducing apoptosis in these cells. In a previous study, we demonstrated that in vitro treatment with perillyl alcohol (POH), an inhibitor of Ras farnezilation, induced apoptosis in human GBM cell lines and explants. Objective: In the presentstudy, we investigated if the partial regression observed in a patient with a recurrent GBM after treatmentby intranasal delivery of POH, is mediated by apoptosis. Result: Data from classical histology, terminaldeoxynucleotidyl-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay, as well asactivation of caspase 3, showed increased apoptosis in the treated tumor. Conclusion: These findings suggest that strategies to inactivate EGFR and RAS signaling may be critical to improving not only theefficacy of single-agent therapy but also of multimodal therapy in gliomas.

Oligodendrogliomas: a genética molecular e o desenvolvimento estratégia terapêutica adjuvante / Oligodendrogliomas: molecular genetics and development of adjuvant therapeutic strategies

A incidência dos oligodendrogliomas tem aumentado provavelmente em razão do progresso na precisão de diagnóstico. Aproximadamente dois terços dos pacientes com a forma mais agressiva, oligodendroglioma anaplásico, mostram resposta substancial à quimioterapia com a associação procarbazina/lomustina/vincristina (PCV). Entretanto, os resultados da quimuiterapia anaplásica dos oligodendrogliomas resulta em grande número de células com ERK/MAPK ativadas. O monoterpeno álcool perílico demonstra atividades quimiopreventiva e quimioterápica em diversos modelos de tumores e sugere-se que estas possam estar associadas com a capacidade de inibir a farmesilação pós-traducional e a sinalização da Ras, assim como a cascata de sinalização por meio da RAF-MEK-ERK. Estudo do nosso grupo observou que pode estar atuando mediante a inibição da fosforilação da extracellular regulated kinase (ERK), uma proteína envolvida na cascata de transdução de sinal através da membrana e proliferação celular induzida pela proteína Ras. Este artigo discute a redução de oligodendroglioma anaplásico recidivado em paciente tratado durante nove meses com álcool perílico por via intranasal.