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This review presents information from several studies that have demonstrated the antiviral activity of extracts ( Andrographis paniculata, Artemisia annua, Artemisia afra, Cannabis sativa, Curcuma longa, Echinacea purpurea, Olea europaea, Piper nigrum, and Punica granatum) and phytocompounds derived from medicinal plants (artemisinins, glycyrrhizin, and phenolic compounds) against SARS-CoV-2. A brief background of the plant products studied, the methodology used to evaluate the antiviral activity, the main findings from the research, and the possible mechanisms of action are presented. These plant products have been shown to impede the adsorption of SARS-CoV-2 to the host cell, and prevent multiplication of the virus post its entry into the host cell. In addition to antiviral activity, the plant products have also been demonstrated to exert an immunomodulatory effect by controlling the excessive release of cytokines, which is commonly associated with SARS-CoV-2 infections.

Produtos naturais, como extratos glicólicos de plantas, são importantes para a aplicação clínica na área da saúde, como em enxaguatórios bucais, cremes dentais e irrigação intracanal. Assim é necessário realizar estudos de citotoxicidade desses extratos glicólicos. O presente estudo buscou avaliar a atividade citotóxica dos extratos glicólicos de Cynara scolymus L. (alcachofra), Myracrodruom urundeuva Allem. (aroeira-do-sertão-do-sertão) e Camellia sinensis (L.) Kuntze (chá verde) em macrófagos de camundongo (RAW 264.7) pelo teste de atividade metabólica MTT. Para tanto, as células foram distribuídas em microplacas de 96 poços e foram expostas a 11 diluições seriadas de cada extrato (200 mg/mL,100 mg/mL, 50 mg/mL, 25 mg/mL, 12,5 mg/mL, 6,25 mg/mL, 3,13 mg/mL, 1,56 mg/mL, 0,78 mg/mL, 0,39 mg/mL e 0,20 mg/mL), sendo n=8 para cada diluição. Após o tempo de contato de 5 minutos e 24 horas, foi avaliada a viabilidade celular utilizando o teste MTT. Diante destes resultados, no tempo de exposição de 5 minutos com os extratos, em ordem crescente de redução da viabilidade celular, seguiram-se o chá verde, com aumento da viabilidade celular, a aroeira-do-sertão e o extrato de alcachofra. Por meio do MTT dos três extratos por tempo de exposição de 24 horas, observou-se que o extrato de alcachofra apresentou maior toxicidade, seguido do extrato de chá verde e aroeira-do-sertão. A análise estatística foi realizada por ANOVA e teste de Tukey, com significância de 5%. Conclusões: Entre os extratos, o chá verde com 12,5mg/mL, com interessantes 5 minutos e 24 horas, despertou atenção, pois foi a maior concentração e não apresentou citotoxicidade para os macrófagos, assim como a alcachofra que foi a mais citotóxica para os macrófagos, em ambos os tempos (5 min e 24 h).

Animals of the primate order are highly susceptible to parasitic infections due to their social organization, often in groups and interaction between individuals, facilitating the transmission of pathogenic agents, and may be even more prevalent in areas modified by anthropic action. The occurrence of the protozoan Cryptosporidium spp. in wildlife animals in captivity or free-living it characterizes great importance of Public Health mainly for its zoonotic potential. This parasite has a cosmopolitan distribution, causes gastrointestinal infection, and its transmission occurs via the fecal-oral route, either through water or through contaminated food. Cryptosporidiosis affects mammals, birds, reptiles and fish, this significant zoonotic potential, in addition to being directly related to the living conditions of the host and the quality of the environment. We investigated the occurrence of Cryptosporidium spp. in primates located at the Tufted Capuchin Monkey Procreation Center (School of Dentistry of Araçatuba - UNESP). Fecal samples from 49 asymptomatic primates of the species Sapajus spp. species were collected below their individual cages, the soil was lined with a plastic bag, thus hindering environmental contamination. The search for oocysts was carried out using the technique of Negative Malachite Green Staining, finding negativity in all samples examined, which suggests the absence of the protozoan Cryptosporidium spp. in primates studied in the breeding center of capuchin monkeys (Sapajus spp.), noting that they were asymptomatic. Los animales del orden de los primates son altamente susceptibles a las infecciones parasitarias debido a su organización social, a menudo en grupos e interacción entre individuos, lo que facilita la transmisión de agentes patógenos, pudiendo ser incluso más prevalentes en áreas modificadas por la acción antrópica. Una ocorrência do protozoário Cryptosporidium spp. em animais selvagens em cativeiro ou de vida livre caracteriza grande importância de Saúde Public principalmente por seu potencial zoonótico. Este parasito apresenta distribuição cosmopolita, causa infecção gastrointestinal e sua transmissão ocorre pela via fecal-oral, seja por via hídrica ou por alimentos contaminados. La criptosporidiosis afecta a mamíferos, aves, reptiles y peces, con un importante potencial zoonótico, además de estar directamente relacionada con las condiciones de vida del huésped y la calidad del medio ambiente. Investigamos la aparición de Crytosporidium spp. en primates ubicados en el Centro de Cría de Monos Capuchinos (Facultad de Odontología de Araçatuba – UNESP). Muestras de heces de 49 primates asintomáticos de la especie Sapajus spp. debajo de sus jaulas individuales, el suelo se cubrió con una bolsa de plástico, lo que impidió la contaminación ambiental. La búsqueda de ooquistes se realizó mediante la Técnica de Tinción Negativa con Verde Malaquita, encontrándose negatividad en todas las muestras examinadas, lo que sugiere la ausencia del protozoario Cryptosporidium spp. en primates estudiados en el centro de cría de monos capuchinos (Sapajus spp.), observándose que eran asintomáticos. Os animais da ordem primata são altamente susceptíveis a infecções parasitárias devido à sua organização social, frequentemente em grupos e interação entre indivíduos, facilitando a transmissão de agentes patogênicos, podendo ser ainda mais prevalentes em áreas modificadas por ação antrópica. A ocorrência do protozoário Cryptosporidium spp. em animais selvagens em cativeiro ou de vida livre caracteriza grande importância de Saúde Pública principalmente por seu potencial zoonótico. Este parasito apresenta distribuição cosmopolita, causa infecção gastrointestinal e sua transmissão ocorre pela via fecal-oral, seja por via hídrica ou por alimentos contaminados. A criptosporidiose afeta mamíferos, aves, répteis e peixes, apresentando significativo potencial zoonótico, além de estar diretamente relacionada às condições de vida do hospedeiro e à qualidade do meio ambiente. Nós investigamos a ocorrência de Cryptosporidium spp. em primatas localizados no Centro de Procriação do Macaco-prego (Faculdade de Odontologia de Araçatuba – UNESP). Amostras fecais de 49 primatas assintomáticos da espécie Sapajus spp. foram coletadas abaixo de suas gaiolas individuais, o solo foi forrado com saco plástico, dificultando assim a contaminação ambiental. A pesquisa de oocistos foi realizada pela técnica de Coloração Negativa de Verde Malaquita, encontrando-se negatividade em todas as amostras examinadas, o que sugere a ausência do protozoário Cryptosporidium spp. nos primatas estudados no núcleo de procriação de macacos-prego (Sapajus spp.), salientando que os mesmos eram assintomáticos.

Objective: The resistance of fungi and bacteria to the available antimicrobials has increased and the development of alternative products to control them has become a very requirement. The use of plant products could be a viable option due to the efficacy, viability, and availability they present. Thereby, this study evaluated the effect of R. officinalis L. extract on C. albicans and S. mutans biofilms, by the total protein level analysis presented by the microorganisms. Material and Methods: For this purpose, monomicrobial biofilms were formed for 48 h and exposed to the R. officinalis L. extract for 5 min. Then, total protein quantification was performed by Lowry method. Results: The analysis showed significant total protein reduction of the biofilms after exposure to the plant extract, with 39 ± 11%, for C. albicans , and 32 ± 11%, for S. mutans. Conclusion: R. officinalis L. extract decreased the total protein level in both biofilms. Thus, C. albicans and S. mutans protein composition could be a target for action of antimicrobial agents. Keywords Biofilms; Candida albicans ; Proteins; Rosmarinus officinalis; Streptococcus mutans.

Made available in DSpace on 2021-06-25T10:59:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01. Added 1 bitstream(s) on 2021-07-15T14:35:57Z : No. of bitstreams: 1 S0001-37652021000300904.pdf: 1633847 bytes, checksum: d62074ab288e57ed75409441fab8da19 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Candida spp. are naturally opportunistic and can promote infections. These yeasts can form biofilm, after penetration and adhesion to the biotic or abiotic surfaces. Preexisting diseases, treatments with drugs and radiation therapy, medical procedures, and parafunctional habits favor the installation of a fungal infection. Increased resistance to the available antifungals has become a concern. Therefore, alternative methods to control them have been evaluated, including the use of plant substances. In this study, the antibiofilm effect of R. officinalis L. extract was analyzed on C. albicans, C. dubliniensis, C. glabrata, C. krusei, and C. tropicalis. A phytochemical analysis of the extract was performed. Biofilms were formed for 48 h and exposed to the different concentrations of the extract (50, 100, and 200 mg/mL) for 5 min or 24 h. The effect of the plant extract was compared to the antifungal nystatin. Rosmarinus officinalis L. extract was constituted of phenols and flavonoids, highlighting the presence of chlorogenic acid derivatives in its composition. Biofilm reductions were observed after exposure to the plant extract for both periods. The plant extract provided a reduction similar to the antifungal. Thus, R. officinalis L. extract showed antibiofilm effect on Candida spp. comparable to the nystatin. Universidade Estadual Paulista(UNESP) Instituto de Ciência e Tecnologia Departamento de Biociências e Diagnóstico Bucal, Av. Engenheiro Francisco José Longo, 777 Universidade Anhembi Morumbi Escola de Medicina, Av. Dep. Benedito Matarazzo, 4050 Universidade Anhanguera Departamento de Farmácia, Av. Raimundo Pereira de Magalhães, 3305 Universidade Federal de São Paulo(UNIFESP) Instituto de Farmacologia e Biologia Molecular Departamento de Modo de Ação de Drogas, Rua Três de Maio, 100 University of Florida College of Dentistry Department of Restorative Dental Sciences Universidade Estadual Paulista(UNESP) Instituto de Ciência e Tecnologia Departamento de Odontologia Restauradora, Av. Engenheiro Francisco José Longo, 777, Universidade Estadual Paulista(UNESP) Instituto de Ciência e Tecnologia Departamento de Biociências e Diagnóstico Bucal, Av. Engenheiro Francisco José Longo, 777 Universidade Estadual Paulista(UNESP) Instituto de Ciência e Tecnologia Departamento de Odontologia Restauradora, Av. Engenheiro Francisco José Longo, 777, FAPESP: 15/08776-3 FAPESP: 15/25338-0

Propolis could represent an alternative therapeutic agent for targeting multidrug-resistant bacteria due to its antimicrobial potential. The effect of Brazilian green propolis (BGP) aqueous extract (AqExt) was evaluated on eight multidrug-resistant clinical strains of

Made available in DSpace on 2021-06-25T10:13:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-09-01 Aim: Plant products have been evaluated to control opportunistic micro-organisms, as well as fortify immune system cells. Thus, Curcuma longa L. (turmeric) extract was evaluated in interactions of murine macrophages (RAW 264.7) with Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans, in order to establish cooperation with defense cells. Materials & methods: Effects of minimal inhibitory concentrations (MIC) of the plant extract were analyzed on phagocytosis, cell viability of RAW 264.7 and production of inflammation-related molecules (IL-1β, TNF-α, IL-10 and NO). Results: The plant extract was cytocompatible and promoted significant reductions of micro-organisms, and synthesis of inflammation-related molecules, during interactions. Conclusion: C. longa L. extract showed significant antimicrobial response and cooperation with macrophages, by fighting bacteria and yeasts during host-microbe interactions. Department of Biosciences and Oral Diagnosis São Paulo State University (UNESP) Institute of Science and Technology, Av. Engenheiro Francisco José Longo, 777 School of Medicine Anhembi Morumbi University, Av. Dep. Benedito Matarazzo, 4050 Anhanguera University, Av. Raimundo Pereira de Magalhães, 3305 Department of Mode of Drug Action Federal University of São Paulo (UNIFESP) Institute of Pharmacology and Molecular Biology, Rua Três de Maio, 100 Department of Restorative Dental Sciences College of Dentistry University of Florida, 1395 Center Drive Department of Biosciences and Oral Diagnosis São Paulo State University (UNESP) Institute of Science and Technology, Av. Engenheiro Francisco José Longo, 777

Made available in DSpace on 2021-06-25T10:12:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Objective: The use of medicinal plants may be an alternative method for the control of Candida spp. responsible for human infections. This study evaluated the antifungal effect of Schinus terebinthifolius extract (Brazilian Peppertree) on C. albicans, C. dubliniensis, C. glabrata, and C. krusei planktonic cultures and biofilms. Material and Methods: Minimum inhibitory concentration (MIC) and minimum fungal concentration (MFC) of the plant extract were determined by the broth microdilution method. Biofilms formed in microplate wells were exposed to the extract for 5 min (50, 100 and 200 mg/mL) or 24 h (25, 50 and 100 mg/mL). After determination of colony-forming units per milliliter (CFU/mL), the data were analyzed by one-way ANOVA and Tukey’s Test (P ≤ 0.05). Results: Different MIC (mg/mL) were found, such as 0.39 (C. dubliniensis), 1.56 (C. albicans), and 3.13 (C. glabrata and C. krusei). Besides, MFC (mg/mL) of 0.78 (C. dubliniensis) and 3.13 (C. albicans, C. glabrata and C. krusei) were also observed. Regarding the biofilms, significant reductions (log10) were found after 5 min and 24 h exposure to the plant extract, compared to the control group. However, C. dubliniensis was significantly affected only in 24 h treatment. Conclusion: S. terebinthifolius extract presented a significant antifungal effect on C. albicans, C. dubliniensis, C. glabrata, and C. Krusei both in planktonic cultures and biofilms. Federal University of Rio de Janeiro (UFRJ) Institute of Biodiversity and Sustainability (NUPEM) University of Taubaté (UNITAU) Basic Institute of Biosciences São Paulo State University (UNESP) Institute of Science and Technology University Center Brazcubas Brazcubas Faculty of Dentistry Anhembi Morumbi University School of Medicine São Paulo State University (UNESP) Institute of Science and Technology

Made available in DSpace on 2018-12-11T17:35:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-15 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) We report the synthesis, optical, electrochemical and electronic properties of a new perylene derivative and its Ru(II) complex, namely N,-(5-amino-1,10-phenantroline)perylene-3,4,9,10-tetracarboximonoimide (pPDI) and Bis(1,10-phenanthroline)(N-(5-amin-1,10-phenantroline)perylene-3,4,9,10-tetracarboxi monoimide)ruthenium(II) hexafluorophosphate, [Ru(phen)2(pPDI)]2+ ([PF6]−)2. The molecular structures of the compounds were elucidated by FTIR, mass spectrometry, elemental analysis (CHN) and DFT calculations. Their optical and electrochemical properties were investigated by absorption and fluorescence spectroscopy and cyclic voltammetry. The spectroscopic and electrochemical studies for both the pPDI and the perylene monoimide metal complex show that Ru2+ coordination does not affect the optical and electrochemical properties of the free perylene bisimide ligand. The Ru complex exhibits emission lifetimes of 5.06 ns (48.1%) and 156 ns (51.9%), resulting from emission of the pPDI ligand and 3MLCT, respectively. The pPDI triplet excited state in the title chromophore is able to sensitize the production of singlet oxygen (1O2). Using a time-resolved laser system, we measured the quantum yield for the production of singlet oxygen (ΦΔ) to be 0.26 for cis-[Ru(phen)2(pPDI)]2+. Candida albicans is recognized as the most common fungal pathogens, causing superficial infections of the skin and mucous membranes. Resistance of C. albicans strains against classical antifungal agents such as fluconazole has increased considerably, which drives the search for new therapeutic alternatives. In the present study, we report that our new Ru-perylene complex can indeed be used to kill the bacterium C. albicans. The fungicidal activity of cis-[Ru(phen)2(pPDI)]2+ against C. albicans was evaluated in the presence of C. albicans cultures in the dark and in the presence of light. After exposure to 12.5 μM of cis-[Ru(phen)2(pPDI)]2 in the presence of light for 180 s, C. albicans showed a decrease of 50% in its concentration compared to the same experimental conditions in the dark. Institute of Exact Sciences and Technology Federal University of Amazonas Department of Chemistry Universidade Federal de São Carlos, CP 676 Department of Biosciences and Oral Diagnosis Universidade Estadual Paulista/UNESP, Av. Engenheiro Francisco José Longo, 777 Department of Chemistry and Biochemistry California State University Department of Biosciences and Oral Diagnosis Universidade Estadual Paulista/UNESP, Av. Engenheiro Francisco José Longo, 777

Stryphnodendron adstringens (Mart.) Coville is a medicinal plant known for its anti-inflammatory and antimicrobial properties. This study evaluated some biological activities of extract from S. adstringens. Antimicrobial activity was checked in planktonic cultures and monomicrobial biofilms on aerobic, and anaerobic dental microorganisms. Analyzes of cytotoxicity using MTT assay, and genotoxicity by micronucleus test were performed in human keratinocytes (HACAT), murine macrophages (RAW 264.7), and murine fibroblasts (L929). The anti-inflammatory effect was checked in RAW 264.7 stimulated by lipopolysaccharide (LPS) from Escherichia coli, and treated with the plant extract. The levels of cytokines, and nitric oxide (NO) were measured by ELISA, and Griess method, respectively. Data were analyzed by ANOVA, followed by Tukey's, or Kruskal-Wallis, and Dunns tests (P ≤ 0.05). Biofilms of anaerobic bacteria were very susceptible to the plant extract. Effective concentrations showed cell viability > 50%, except 25 mg/mL for HACAT after 24 h of exposure. The extract of S. adstringens was not genotoxic for RAW 264.7. LPS associated with extract increased the production of all cytokines, except TNF-α. However, the plant extract decreased the production of NO. In conclusion, the extract of S. adstringens affected biofilm of anaerobic bacteria using non-cytotoxic concentrations for RAW 264.7, L929, and HACAT cells. Stryphnodendron adstringens (Mart.) Coville es una planta medicinal conocida por sus propiedades antiinflamatorias y antimicrobianas. Este estudio evaluó algunas actividades biológicas del extracto de S. adstringens. La actividad antimicrobiana se verificó en cultivos planctónicos y biopelícula monomicrobianos en microorganismos aerobios y anaerobios de interés dental. Se realizaron análisis de citotoxicidad mediante ensayo MTT y genotoxicidad mediante ensayo de micronúcleos en queratinocitos humanos (HACAT), macrófagos murinos (RAW 264.7) y fibroblastos murinos (L929). El efecto antiinflamatorio se verificó en RAW 264.7 estimulado por lipopolisacárido (LPS) de Escherichia coli y tratado con el extracto vegetal. Los niveles de citocinas y óxido nítrico (NO) se midieron mediante ELISA y el método de Griess, respectivamente. Los datos se analizaron mediante ANOVA y prueba de Tukey o prueba de Kruskal-Wallis y Dunns (P ≤ 0,05). Las biopelículas de bacterias anaerobias fueron muy susceptibles al extracto de la planta. Las concentraciones efectivas mostraron viabilidad celular> 50%, excepto 25 mg/mL para HACAT después de 24 h de exposición. El extracto de S. adstringens no fue genotóxico para RAW 264.7. El LPS asociado con el extracto aumentó la producción de todas las citocinas, excepto TNF-α. Sin embargo, el extracto de la planta disminuyó la producción de NO. En conclusión, el extracto de S. adstringens afectó el biopelícula de bacterias anaerobias utilizando concentraciones no citotóxicas para RAW 264.7, L929 y HACAT. Stryphnodendron adstringens (Mart.) Coville é uma planta medicinal conhecida por suas propriedades anti-inflamatórias e antimicrobianas. Este estudo avaliou algumas atividades biológicas do extrato de S. adstringens. A atividade antimicrobiana foi verificada em culturas planctônicas e biofilmes monomicrobianos em microrganismos aeróbios e anaeróbios de interesse odontológico. Análises de citotoxicidade utilizando ensaio de MTT e genotoxicidade por meio do teste de micronúcleo foram realizadas em queratinócitos humanos (HACAT), macrófagos murinos (RAW 264.7) e fibroblastos murinos (L929). O efeito anti-inflamatório foi verificado em RAW 264.7 estimulado por lipopolissacarídeo (LPS) de Escherichia coli e tratado com o extrato da planta. Os níveis de citocinas e óxido nítrico (NO) foram medidos por ELISA e método de Griess, respectivamente. Os dados foram analisados ​​por ANOVA e teste de Tukey ou Kruskal-Wallis e teste de Dunns (P ≤ 0,05). Os biofilmes de bactérias anaeróbias foram muito suscetíveis ao extrato da planta. As concentrações efetivas mostraram viabilidade celular > 50%, exceto 25 mg/mL para HACAT após 24 h de exposição. O extrato de S. adstringens não foi genotóxico para RAW 264.7. O LPS associado ao extrato aumentou a produção de todas as citocinas, exceto TNF-α. No entanto, o extrato da planta diminuiu a produção de NO. Em conclusão, o extrato de S. adstringens afetou o biofilme de bactérias anaeróbias usando concentrações não citotóxicas para células RAW 264.7, L929, e HACAT.

Made available in DSpace on 2018-12-11T17:13:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-08-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Abstract: Microorganisms are capable to combat defense cells by means of strategies that contribute to their stabilization and proliferation in invaded tissues. Frequently antimicrobial-resistant strains appear; therefore, alternative methods to control them must be investigated, for example, the use of plant products. The capacity of the thyme extract (Thymus vulgaris L.) and phytocompound thymol in the control of in vitro infections by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in murine macrophages (RAW 264.7) was evaluated. Minimal inhibitory concentrations (MIC) of the plant products were used. The effect of these MIC were analyzed in the assays of phagocytosis and immunoregulation by analysis of the production of cytokines (IL-1β, TNF-α, and IL-10) and nitric oxide (NO). The plant products effectively assisted the macrophages in the phagocytosis of microorganisms, presenting significant reductions of S. aureus and P. aeruginosa. The macrophages also regulated the production of inflammatory mediators in the infections by S. aureus, P. aeruginosa, and C. albicans. In addition, thyme provided a satisfactory effect in response to the bacterial infections, regarding generation of NO. Thus, the effectiveness of the thyme and thymol to control in vitro infections by S. aureus, P. aeruginosa, and C. albicans was observed. Highlights: Phagocytosis of S. aureus by RAW 264.7 was enhanced with thymolThyme enhanced the phagocytosis of P. aeruginosa by RAW 264.7Plant products provided immunoregulation of inflammatory cytokinesProduction of nitric oxide was improved with the treatments in bacterial infections. Department of Biosciences and Oral Diagnosis São Paulo State University (UNESP)—Institute of Science and Technology Departamento de Biociências e Diagnóstico Bucal Laboratório de Microbiologia e Imunologia Universidade Estadual Paulista/UNESP—Instituto de Ciência e Tecnologia, Av. Engenheiro Francisco José Longo, 777—Jardim São Dimas Department of Biosciences and Oral Diagnosis São Paulo State University (UNESP)—Institute of Science and Technology Departamento de Biociências e Diagnóstico Bucal Laboratório de Microbiologia e Imunologia Universidade Estadual Paulista/UNESP—Instituto de Ciência e Tecnologia, Av. Engenheiro Francisco José Longo, 777—Jardim São Dimas FAPESP: 2015/08776-3

Objective: This study evaluated the effect of rosemary extract on Candida albicans , Staphylococcus aureus , Enterococcus faecalis , Streptococcus mutans and Pseudomonas aeruginosa monomicrobial biofilms viability, as well as on C. albicans associated with S. aureus , E. faecalis , S. mutans or P. aeruginosa in polymicrobial biofilms. Material and Methods: In microtiter plate, mono- and polymicrobial biofilms for 48 h were formed. Then, they were exposed for 5 min to rosemary extract (200 mg/mL). Saline (0.9% NaCl) was used as control. After, washes were done with saline to remove the affected cells. Biofilm viability was checked by MTT colorimetric assay, after treatment. Absorbance of the wells was read in microplate spectrophotometer (570 nm) and data were converted to reduction percentage and statistically analyzed by ANOVA and Tukey test ( p ≤ 0.05). Results: After application of rosemary extract, with exception of the E. faecalis biofilm, significant reductions in mono- and polymicrobial biofilms viability were observed. Conclusion: C. albicans , S. aureus , S. mutans and P. aeruginosa monomicrobial biofilms were affected by rosemary extract, as well as C. albicans associated with S. aureus , E. faecalis , S. mutans or P. aeruginosa in polymicrobial biofilms, presenting significant viability reductions.

Made available in DSpace on 2020-12-12T01:27:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-01 Finding an effective alternative way to aid defense cells to fight Streptococcus mutans was the main goal of this study. The effect of plant extracts from Curcuma longa L. (turmeric), Rosmarinus officinalis L. (rosemary), and Thymus vulgaris L. (thyme) was evaluated on murine macrophages (RAW 264.7) infected by S. mutans. Minimum inhibitory concentration (MIC) of the extracts was determined. Macrophages were infected by S. mutans and treated with each extract. From the supernatants, it was measured nitric oxide (NO) level. Posteriorly, RAW 264.7 were lysed to expose living and phagocytosed bacteria. Cytotoxicity was checked by lysosomal activity analysis, using neutral red assay. Each extract helped RAW 264.7 to eliminate S. mutans during infection, as observed by a significant bacterial reduction. Significant cell viability was also found. Besides, an increased production of NO was verified using R. officinalis L. and T. vulgaris L. extracts. The evaluated extracts demonstrated an effective action to assist RAW 264.7 to fight S. mutans during infection. Department of Biosciences and Oral Diagnosis Institute of Science and Technology São Paulo State University (UNESP), Av. Engenheiro Francisco José Longo, 777, Jardim São Dimas Anhembi Morumbi University School of Medicine, Av. Dep. Benedito Matarazzo, 4050 Department of Restorative Dental Sciences College of Dentistry University of Florida Department of Biosciences and Oral Diagnosis Institute of Science and Technology São Paulo State University (UNESP), Av. Engenheiro Francisco José Longo, 777, Jardim São Dimas

Made available in DSpace on 2020-12-12T02:26:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-09-01 Biological effects of titanium (Ti) alloys were analyzed on biofilms of Candida albicans, Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus mutans, and Streptococcus sanguinis, as well as on osteoblast-like cells (MG63) and murine macrophages (RAW 264.7). Standard samples composed of aluminum and vanadium (Ti-6Al-4V), and sample containing niobium (Ti-35Nb) and zirconium (Ti-13Nb-13Zr) were analyzed. Monomicrobial biofilms were formed on the Ti alloys. MG63 cells were grown with the alloys and the biocompatibility (MTT), total protein (TP) level, alkaline phosphatase (ALP) activity, and mineralization nodules (MN) formation were verified. Levels of interleukins (IL-1β and IL-17), tumor necrosis factor alpha (TNF-α), and oxide nitric (NO) were checked, from RAW 264.7 cells supernatants. Data were statically analyzed by one-way analysis of variance (ANOVA) and Tukey’s test, or T-test (P ≤ 0.05). Concerning the biofilm formation, Ti-13Nb-13Zr alloy showed the best inhibitory effect on E. faecalis, P. aeruginosa, and S. aureus. And, it also acted similarly to the Ti-6Al-4V alloy on C. albicans and Streptococcus spp. Both alloys were biocompatible and similar to the Ti-6Al-4V alloy. Additionally, Ti-13Nb-13Zr alloy was more effective for cell differentiation, as observed in the assays of ALP and MN. Regarding the stimulation for release of IL-1β and TNF-α, Ti-35Nb and Ti-13Nb-13Zr alloys inhibited similarly the synthesis of these molecules. However, both alloys stimulated the production of IL-17. Additionally, all Ti alloys showed the same effect for NO generation. Thus, Ti-13Nb-13Zr alloy was the most effective for inhibition of biofilm formation, cell differentiation, and stimulation for release of immune mediators. [Figure not available: see fulltext.]. Department of Biosciences and Oral Diagnosis Institute of Science and Technology São Paulo State University (UNESP), Av. Engenheiro Francisco José Longo, 777 Division of Materials Air and Space Institute (CTA) Department of Materials and Dental Prosthodontics Institute of Science and Technology São Paulo State University (UNESP), Av. Engenheiro Francisco José Longo, 777 Brazcubas Faculty of Dentistry University Center Brazcubas, Av. Francisco Rodrigues Filho, 1233 Department of Biosciences and Oral Diagnosis Institute of Science and Technology São Paulo State University (UNESP), Av. Engenheiro Francisco José Longo, 777 Department of Materials and Dental Prosthodontics Institute of Science and Technology São Paulo State University (UNESP), Av. Engenheiro Francisco José Longo, 777

The use of medicinal plants can be an alternative method for the control of microorganisms responsible for human infections. This study evaluated the antimicrobial activity of Salvia officinalis Linnaeus (sage) extract on clinical samples isolated from the oral cavity and reference strains of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Candida albicans, Candida tropicalis, and Candida glabrata. In addition, testing assessed the cytotoxic effect of S officinalis on murine macrophages (RAW 264.7). Minimum inhibitory, minimum bactericidal, and minimum fungicidal concentrations of S officinalis extract were determined by broth microdilution method in 60 microbial samples. The cytotoxicity was checked by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The quantities of the proinflammatory cytokines interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) produced by RAW 264.7 were analyzed by an enzyme-linked immunosorbent assay. An S officinalis concentration of 50.0 mg/mL was effective against all microorganisms. Regarding cytotoxicity, the groups treated with 50.0-, 25.0-, and 12.5-mg/mL concentrations of S officinalis presented cell viability statistically similar to that of the control group, which was 100% viable. The production of IL-1β and TNF-α was inhibited at a 50.0-mg/mL concentration of S officinalis. Thus, S officinalis extract presented antimicrobial activity on all isolates of Staphylococcus spp, S mutans, and Candida spp. No cytotoxic effect was observed, as demonstrated by the survival of RAW 264.7 and inhibition of IL-1β and of TNF-α.

Made available in DSpace on 2019-10-04T12:32:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01. Added 1 bitstream(s) on 2021-07-15T15:22:58Z : No. of bitstreams: 1 S1678-77572019000100404.pdf: 2187469 bytes, checksum: 25307a0ef1d72e3d6a3429ca5beadba2 (MD5) Objective: Myofibroblasts have been associated with the development of several pathologic fibrotic conditions. This longitudinal study aims to assess the proliferative and antiapoptotic effects of cyclosporin, nifedipine and phenytoin on gingival connective tissue cells of nonhuman primate, as well as to analyze a possible role of myofibroblasts in gingival overgrowth. Materials and Methods: Gingival samples from the right superior canine area were obtained from 12 male monkeys (Sapajus spp) to comprise the control group. After one week, the animals were randomly assigned to three groups, which received daily oral doses of cyclosporin, nifedipine or phenytoin for 120 days. Gingival samples were collected from the left superior canine area of two animals of each group at 52 and 120 days. Histological sections were stained with hematoxylin and eosin, and immunoreacted against alpha-SMA, Ki67 and bcl-2. Results: alpha-SMA immunoreaction was negative in the control and experimental groups. Similarly, no difference between groups concerning immunostaining against Ki-67 and bcl-2 was observed in connective tissue cells. Conclusion: Based on this methodology, it may be concluded that gingival overgrowths induced by cyclosporin, nifedipine and phenytoin are not associated with neither myofibroblast transdifferentiation, proliferation nor apoptosis of gingival connective cells in monkeys. Univ Estadual Paulista, Fac Odontol, Dept Emergencia, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Odontol, Dept Ciencias Basicas, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Odontol, Dept Patol, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Odontol, Propedeut Clin, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Odontol, Dept Emergencia, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Odontol, Dept Ciencias Basicas, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Odontol, Dept Patol, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Odontol, Propedeut Clin, Aracatuba, SP, Brazil

8º Congresso de Odontologia de Araçatuba, 38º Jornada Acadêmica: Prof. Dr. Mauro Airton Rulli, 14º Simpósio Pós-Graduação Profa. Dra. Kikue Takebayashi Sassaki, 10º Encontro do CAOE, Encontro de 40 anos de Egressos de Pós-Graduação em Odontologia (Área de C.T.B-M-F)

Candida species inhabit the oral cavity of all individuals who wear complete denture and whose material is the same as that used in splints. Assess the growth of C. albicans in occlusal and palatal splints used for treatment of TMD so that the potential risks of oral microbiota can be assessed. The growth of Candida spp. was assessed in the saliva of 27 individuals wearing splints for treatment of TMD. They were divided into two groups: G1 (n = 14), individuals wearing occlusal splint; and G2 (n = 13), individuals wearing palatal splint. Saliva samples were collected during placement of the splints (T1) and after 4 months (T2), being stored in PBS (10 mL) after 60-second rinses. It was observed that patients wearing occlusal splints (G1) had an increase of 0.648 CFU/mL (Log 10), with statistically significant differences (P = 0.043) for C. albicans (42.33%), C. glabrata (5.52%), C. krusei (41.72%) and C. tropicalis (10.43%). In the group of patients wearing palatal splints (G2), there was a decrease of 0.101 CFU/mL (Log 10), was observed with (P = 0.964) only the presence of C. albicans. The results suggest that growth of Candida species was greater in patients wearing occlusal splints compared to those wearing palatal ones as the presence of different yeast species was found in the former. Espécies de Candida habitam a cavidade oral de 60-100% de indivíduos usuários de prótese total, cujo material é o mesmo utilizado em placas miorrelaxante. Avaliar o crescimento de C. albicans. em placas relaxantes musculares oclusais e palatais, usadas para o tratamento de DTM, na intenção de verificar riscos em potencial à microbiota bucal. Avaliou-se o crescimento de Candida spp. na saliva de 27 indivíduos, usuários de placa miorrelaxante, em tratamento para DTM no ICT-UNESP. Os indivíduos foram divididos em dois grupos: G1(n=14) – placa com recobrimento oclusal; e G2 (n=13) – sem recobrimento. As coletas foram com PBS (10mL), em bochechos por 60seg, na instalação das placas (T1) e após 4 meses (T2). Observou-se que pacientes usuários da placa miorrelaxante com recobrimento oclusal (grupo G1) apresentaram aumento de 0,648 UFC/mL (Log10) com diferença estatisticamente significante (p=0,043) analisando-se 42,33% C. albicans, 5,52% C. glabrata, 41,72% C. krusei e 10,43% C. tropicalis. No grupo de pacientes que utilizaram a placa sem recobrimento (grupo G2), observou-se diminuição de 0,101 UFC/mL (Log10) com (p=0,954) apresentando apenas C. albicans. Os resultados sugerem que os pacientes que fizeram uso de placa miorrelaxante com recobrimento oclusal apresentaram maior crescimento de Candida spp. em relação aos usuários de placa sem recobrimento, verificando-se a presença de diferentes espécies da levedura.

This study evaluated the in vitro antibiofilm effect of 5 different commercial mouthwashes (Cepacol Traditional, Colgate Plax Fresh Mint, Listerine Cool Mint, Oral-B Complete, and Sensodyne) on Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans, Escherichia coli, and Pseudomonas aeruginosa. The cytotoxic effect of the mouthwashes on gingival fibroblasts was also analyzed. A colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used to investigate the viability of biofilms after 48 hours and gingival fibroblasts after 24 hours. The biofilms were exposed to the mouthwashes for 2 different lengths of time: T1, the time recommended by the manufacturer (30 or 60 seconds); and T2, double the recommended time (60 or 120 seconds). All antiseptic mouthwashes caused a significant reduction of biofilm (P0.05) as well as a significant reduction of viable gingival fibroblasts (P0.05) with both exposure times (T1 and T2). It can be concluded that the commercial mouthwashes demonstrated effective antibiofilm activity; they were more effective on bacteria than on C albicans. A significant cytotoxic effect on gingival fibroblasts was also observed.

The aims of this study were evaluate cytotoxicity, genotoxicity, antimicrobial activity of desensitizing toothpastes compared to a common one and the surface roughness of tooth enamel submitted to brushing with these toothpastes. Samples of three desensitizing toothpastes (Colgate Sensitive, Sensodyne and Oral B Sensitive) and common toothpaste (Colgate) were placed in contact with gingival human fibroblasts. Cytotoxicity and genotoxocity were measured by MTT assay and micronucleus test. Antimicrobial activity of the toothpastes extracts against C. albicans, S. mutans and S. aureus were assessed. For surface roughness evaluation, bovine teeth were submitted to 10.000 brushing cycles. The results were analyzed statically using Mann-Whitney U, ANOVA and Z tests (p0.05). Based on these results, we can conclude that although none toothpaste has induced genotoxicity, Colgate Sensitive was also not cytotoxic. Colgate was the most effective against the microorganisms, and there were no differences on the enamel surface roughness between the groups. Resumo Os objetivos desse estudo foram avaliar a citotoxicidade, genotoxicidade, atividade antimicrobiana de dentifrícios dessensibilizantes em comparação com um comum e também a rugosidade superficial do esmalte dentário submetido à escovação com esses dentifrícios. Amostras de três dentifrícios dessensibilizantes (Colgate Sensitive, Sensodyne e Oral B Sensitive) e um dentifrício comum (Colgate) foram colocadas em contato com fibroblastos gengivais humanos e a citotoxicidade e genotoxidade foram mensuradas pelo ensaio MTT e teste do micronúcleo. A atividade antimicrobiana dos extratos dos dentifrícios contra C. albicans, S. mutans e S. aureus foi determinada. Para a avaliação da rugosidade superficial, espécimes de dentes bovinos foram submetidas à 10.000 ciclos de escovação. Os resultados foram analisados estatisticamente usando os testes Mann-Whitney U, ANOVA e Teste Z (P0,05). Com base nesses resultados, podemos concluir que, embora nenhum dentifrício tenha induzido genotoxicidade, o Colgate Sensitive também não foi citotóxico. O Colgate foi o mais eficaz contra os microorganismos, e não houve diferença na rugosidade superficial do esmalte entre os grupos.

Made available in DSpace on 2018-11-12T17:29:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-06-01. Added 1 bitstream(s) on 2018-11-12T17:37:17Z : No. of bitstreams: 1 S1981-86372017000200148.pdf: 63905 bytes, checksum: 3b2dc3c60ddec54577eebc4429a047ca (MD5) RESUMO Fitoterápicos são produtos derivados de plantas com propriedades medicinais. Eles são utilizados para o tratamento ou a prevenção de diversas doenças. No entanto, os pacientes que fazem uso destas substâncias, na sua grande maioria, desconhecem seus efeitos negativos. Portanto, o objetivo deste estudo foi realizar uma breve revisão de literatura sobre os efeitos anticoagulantes de fitoterápicos e sua importância diante da realização de procedimentos odontológicos cirúrgicos. Medidas de prevenção, esclarecimento e acompanhamento de pacientes que se utilizam destes medicamentos são recomendados antes da realização destes procedimentos a fim de prevenir complicações como processos hemorrágicos. ABSTRACT Phytotherapeutic drugs are plant-derived products with medicinal properties. They are used for treating or preventing several diseases. However, patients who use these substances and even health professionals are unaware of their negative effects. One of the most common negative effects of phytotherapeutic drugs reported in the literature is the inhibition of natural coagulation factors in the human body. Therefore, the aim of this study was to perform a brief review of the literature concerning the anticoagulant effects of phytotherapeutic drugs and their importance in surgical dental procedures. Preventive measures, clarification and monitoring of patients taking phytotherapeutic drugs are recommended before performing surgical dental procedures, in order to prevent against complications such as hemorrhage. Prática Clínica Limitada à Endodontia Universidade Federal de Alfenas Faculdade de Odontologia Departamento de Odontologia Restauradora Universidade Paranaense Faculdade de Odontologia Departamento de Diagnóstico Oral Universidade Estadual de São Paulo Instituto de Ciência e Tecnologia Departamento de Biopatologia Oral Universidade Estadual de São Paulo Instituto de Ciência e Tecnologia Departamento de Odontologia Restauradora

Made available in DSpace on 2018-12-11T16:48:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-12-01 Recently, the development of coatings to protect biomedical alloys from oxidation, passivation and to reduce the ability for a bacterial biofilm to form after implantation has emerged. Diamond-like carbon films are commonly used for implanted medical due to their physical and chemical characteristics, showing good interactions with the biological environment. However, these properties can be significantly improved when titanium dioxide nanoparticles are included, especially to enhance the bactericidal properties of the films. So far, the deposition of hydroxyapatite on the film surface has been studied in order to improve biocompatibility and bioactive behavior. Herein, we developed a new route to obtain a homogeneous and crystalline apatite coating on diamond-like carbon films grown on 304 biomedical stainless steel and evaluated its antibacterial effect. For this purpose, films containing two different concentrations of titanium dioxide (0.1 and 0.3 g/L) were obtained by chemical vapor deposition. To obtain the apatite layer, the samples were soaked in simulated body fluid solution for up to 21 days. The antibacterial activity of the films was evaluated by bacterial eradication tests using Staphylococcus aureus biofilm. Scanning electron microscopy, X-ray diffraction, Raman scattering spectroscopy, and goniometry showed that homogeneous, crystalline, and hydrophilic apatite films were formed independently of the titanium dioxide concentration. Interestingly, the diamond-like films containing titanium dioxide and hydroxyapatite reduced the biofilm formation compared to controls. A synergism between hydroxyapatite and titanium dioxide that provided an antimicrobial effect against opportunistic pathogens was clearly observed. Laboratory of Biomedical Nanotechnology Universidade Brasil Laboratory of Biomedical Nanotechnology Universidade do Vale do Paraiba Department of Biosciences and Oral Diagnosis Institute of Science and Technology UNESP-Univ Estadual Paulista Associated Laboratory of Sensors and Materials National Institute for Space Research Biomaterials Innovation Research Center Department of Medicine Brigham and Women's Hospital Harvard Medical School Nanomedicine Lab Department of Chemical Engineering Northeastern University Interdisciplinary Laboratory for Advanced Materials PPGCM Technology Center Federal University of Piaui Department of Biosciences and Oral Diagnosis Institute of Science and Technology UNESP-Univ Estadual Paulista

Made available in DSpace on 2018-11-26T17:24:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-03-01 Introduction: Nonsyndromic primary failure of eruption (PFE) is a rare autosomal dominant disorder of dental eruption with no obvious dental or soft tissue interference. The purposes of this study were to genetically and clinically characterize a family with many members affected by PFE and to describe the natural evolution of the disorder. Methods: Three generations of a family with 18 members, 10 of them clinically affected by PFE, were evaluated periodically during 20 years of clinical follow-up. PFE was observed in varying degrees of severity in both sexes. Clinical presentation became more severe in adulthood. One patient had spontaneous reeruption of 2 posterior teeth. Cervical root resorptions were observed in 3 members. Genetic analysis showed a deleterious heterozygous mutation in intron 9 of the PTH1R gene (c.639-2A>G) and diagnosed an additional affected member. Conclusions: The long-term follow-up of PFE cases in this family permitted the following observations: (1) the onset occurred from the preemergence to the postemergence phases, (2) PFE appeared to be closely related to ankylosis, (3) affected teeth maintained the eruptive potential even in adulthood, (4) the earlier the onset the more severe the open bite, and (5) cervical root resorptions occurred in 3 affected members. Univ Estadual Paulista, Aracatuba Dent Sch, Emergency Dept, Aracatuba, SP, Brazil Univ Estadual Paulista, Aracatuba Dent Sch, Dept Basic Sci, Div Anat, Aracatuba, SP, Brazil Univ Estadual Paulista, Vet Med Sch Aracatuba, Lab Anim Biochem & Mol Biol, Aracatuba, SP, Brazil Univ Regensburg, Inst Human Genet, Regensburg, Germany Univ Estadual Paulista, Aracatuba Dent Sch, Emergency Dept, Aracatuba, SP, Brazil Univ Estadual Paulista, Aracatuba Dent Sch, Dept Basic Sci, Div Anat, Aracatuba, SP, Brazil Univ Estadual Paulista, Vet Med Sch Aracatuba, Lab Anim Biochem & Mol Biol, Aracatuba, SP, Brazil

É de grande importância ampliar as pesquisas científicas com plantas medicinais sobre micro-organismos. Foi verificado o efeito antimicrobiano do extrato de C. sinensis , sobre cultura planctônica e biofilme de S. aureus. Neste estudo foi verificada ação do extrato entre 50 a 0,09 mg/mL sobre cultura planctônica, para determinação da CIM e CBM. A ação do extrato sobre biofilme foi analisada pela exposição de 5 min ao produto vegetal a 200mg/mL. Foi utilizado o método de MTT para determinar a redução da viabilidade do biofilme. Nos grupos controles foram utilizadas clorexidina (+) e solução salina (-), sendo n = 12/grupo. A redução do biofilme foi analisada por ANOVA e Tukey Test (p ≤ 0,05). Em cultura planctônica as concentrações avaliadas não foram efetivas, contudo, utilizando-se a concentração de 200 mg/mL sobre o biofilme, foi verificada redução significativa. Com isso, concluiu-se que o extrato de chá verde demonstrou ação sobre biofilme de S. aureus provendo redução significativa em sua viabilidade.

Alcachofra é um vegetal utilizado na culinária e algumas de suas ações biológicas foram descritas, como ação antioxidante e anti-inflamatória, porém foi pouco relatada sua atividade sobre micro-organismos. Com isso, o objetivo do presente estudo foi verificar o efeito antimicrobiano do extrato de alcachofra sobre P. aeruginosa. Primeiramente, foi verificada a ação deste extrato sobre cultura planctônica, pelo método de microdiluição em caldo (50 a 0,09 mg/mL), em seguida, sobre biofilme formado por 48 h em poços de placas de microtitulação, com exposição de 5 min ao extrato de alcachofra (200 mg/mL). Solução salina 0,9% e clorexidina 0,12% foram utilizadas como controles. As reduções do biofilme foram verificadas pelo teste de MTT e analisadas estatisticamente por ANOVA e Tukey Test (p ≤ 0,05). Em culturas planctônicas foi verificado que as concentrações avaliadas não foram efetivas (CIM > 50 mg/mL), contudo, sobre o biofilme foi verificada redução significativa de 42 ± 16%. Desta forma, pode-se concluir que o extrato de alcachofra apresentou efeito antimicrobiano sobre biofilme de P. aeruginosa.

Made available in DSpace on 2018-12-11T17:10:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-03-01 R. officinalis L. is an aromatic plant commonly used as condiment and for medicinal purposes. Biological activities of its extract were evaluated in this study, as antimicrobial effect on mono- and polymicrobial biofilms, cytotoxicity, anti-inflammatory capacity, and genotoxicity. Monomicrobial biofilms of Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans and Pseudomonas aeruginosa and polymicrobial biofilms composed of C. albicans with each bacterium were formed in microplates during 48 h and exposed for 5 min to R. officinalis L. extract (200 mg/mL). Its cytotoxic effect was examined on murine macrophages (RAW 264.7), human gingival fibroblasts (FMM-1), human breast carcinoma cells (MCF-7), and cervical carcinoma cells (HeLa) after exposure to different concentrations of the extract, analyzed by MTT, neutral red (NR), and crystal violet (CV) assays. The anti-inflammatory activity was evaluated on RAW 264.7 non-stimulated or stimulated by lipopolysaccharide (LPS) from Escherichia coli and treated with different concentrations of the extract for 24 h. Interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) were quantified by ELISA. Genotoxicity was verified by the frequency of micronuclei (MN) at 1000 cells after exposure to concentrations of the extract for 24 h. Data were analyzed by T-Test or ANOVA and Tukey Test (P ≤ 0.05). Thus, significant reductions in colony forming units per milliliter (CFU/mL) were observed in all biofilms. Regarding the cells, it was observed that concentrations ≤ 50 mg/mL provided cell viability of above 50%. Production of proinflammatory cytokines in the treated groups was similar or lower compared to the control group. The MN frequency in the groups exposed to extract was similar or less than the untreated group. It was shown that R. officinalis L. extract was effective on mono- and polymicrobial biofilms; it also provided cell viability of above 50% (at ≤ 50 mg/mL), showed anti-inflammatory effect, and was not genotoxic. Impact statement: Rosmarinus officinalis L. extract effectively contributed to in vitro control of important species of microorganisms such as Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans, and Pseudomonas aeruginosa in mono- and polymicrobial biofilms that are responsible for several infections in oral cavity as in other regions of the body. Furthermore, this extract promoted also cell viability above 50% at concentrations ≤ 50 mg/mL, excellent anti-inflammatory effect, showing inhibition or reduction of the synthesis of proinflammatory cytokines, being also non-genotoxic to cell lines studied. Thus, this extract may be a promising therapeutic agent that can be added in some medical and dental formulations such as toothpastes, mouthwashes, irrigating root canals, ointments, soaps, in order to control pathogenic microorganisms and biofilms, with anti-inflammatory effect and absence of cytotoxic and genotoxic. Department of Biosciences and Oral Diagnosis Institute of Science and Technology Univ Estadual Paulista/UNESP, São José dos Campos, SP Institute of Research and Development Universidade do Vale do Paraíba/UNIVAP, São José dos Campos, SP Department of Biosciences and Oral Diagnosis Institute of Science and Technology Univ Estadual Paulista/UNESP, São José dos Campos, SP

Objective: Evaluating the antifungal potential of Equisetum arvense L. (horsetail), Glycyrrhiza glabra L. (licorice), Punica granatum L. (pomegranate) and Stryphnodendron barbatimam Mart. (barbatimao) extracts, after Candida albicans biofilm formation on acrylic resin. Material and methods: C. albicans standard strain was cultured on Sabourauddextrose agar for 24 h at 37 °C. After standardized in a spectrophotometer, 100 μL of the inoculum (106 cells/mL) and a sterile acrylic resin disc were maintained in Brain Heart Infusion broth supplemented with sucrose (5%), for 5 days at 37 oC. The samples of the treated groups (n = 10) were separately exposed to a concentration of 50 mg/mL of each extract for 5 minutes or to nystatin (48.83 IU/mL). For the untreated group (control, n = 10), it was used sterile saline (0.9% NaCl). Biofilms were disaggregated from the acrylic resin discs by an ultrasonic homogenizer for 30 s. After decimal dilutions, sowings in Sabouraud-dextrose plates were made with incubation for 48 h at 37°C. Later, CFU/mL was verified and the values were converted to log10 and they had their statistical analysis done (ANOVA and Tukey Test, p ≤ 0.05). Results: It was found that all plant extracts and nystatin resulted in significant reduction of C. albicans biofilm (p < 0.01) compared to the control group (0.9% NaCl). However, all of them showed similar reductions to each other (p = 0.1567). Conclusion: There was biofilm formation of C. albicans on acrylic resin and all plant extracts were effective against this yeast, acting similarly to nystatin.

Made available in DSpace on 2018-12-11T17:33:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Objectives This study evaluated the biological effects of the T. vulgaris L. extract., such as antimicrobial activity on planktonic cultures and mono- and polymicrobial biofilms, cytotoxicity, anti-inflammatory activity and genotoxicity. Methods Monomicrobial biofilms of Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans and Pseudomonas aeruginosa and polymicrobial biofilms composed by C. albicans with each bacterium were formed for 48 h and exposed for 5 min to the plant extract. Murine macrophages (RAW 264.7), human gingival fibroblasts (FMM-1), human breast carcinoma cells (MCF-7) and cervical carcinoma cells (HeLa) were also exposed to the plant extract for 5 min and the cell viability were analyzed by MTT, neutral red (NR) and crystal violet (CV) assays. Interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) produced by RAW 264.7 was quantified by ELISA, after 24 h exposure to the plant extract, both in the absence and presence of lipopolysaccharide (LPS) from Escherichia coli. Genotoxicity of the plant extract was evaluated by micronucleus formation (MN) in 1000 cells. The results were analyzed by T-Test or ANOVA and Tukey's Test (P ≤ 0.05). Results All biofilms showed significant reductions in CFU/mL (colony-forming units per milliliter). Cell viability was above 50% for all cell lines. Anti-inflammatory effect on the synthesis of IL-1β and TNF-α was observed. The MN was similar or lower than the control group in all cells. Conclusions T. vulgaris L. extract was effective against all biofilms, promoted high cell viability, anti-inflammatory effect and presented no genotoxicity. São Paulo State University (UNESP) Institute of Science and Technology Department of Biosciences and Oral Diagnosis Universidade do Vale do Paraíba (UNIVAP) Institute of Research and Development São Paulo State University (UNESP) Institute of Science and Technology Department of Biosciences and Oral Diagnosis FAPESP: 2015/08776-3

We present a study of Per3 expression in six different tissues of the non-human primate Cebus apella (capuchin monkey). The aim of this study was to verify whether the expression of the Per3 gene in different tissues of capuchin monkey occurs in a circadian pattern, its phase and the phase relationships between these different tissues during the 24 h of a day. We observed that gene expression oscillated in all of the tissues studied during this time period, although only the liver and muscle presented a robust circadian pattern. This preliminary study highlights the possibility of using Cebus apella as a model to study circadian rhythms at the gene expression level and opens an opportunity for future researches.

Univ Estadual Paulista UNESP, Sch Dent Aracatuba, Dept Surg & Integrated Clin, Aracatuba, Brazil

Made available in DSpace on 2016-01-28T16:56:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2011. Added 1 bitstream(s) on 2016-01-28T17:02:54Z : No. of bitstreams: 1 ISSN1808-8694-2011-77-04-439-446-en.pdf: 3175665 bytes, checksum: 1a6e033536e2f48753c288729ca206c1 (MD5). Added 1 bitstream(s) on 2016-01-28T17:02:55Z : No. of bitstreams: 2 ISSN1808-8694-2011-77-04-439-446-en.pdf: 3175665 bytes, checksum: 1a6e033536e2f48753c288729ca206c1 (MD5) ISSN1808-8694-2011-77-04-439-446-pt.pdf: 3178513 bytes, checksum: 887962c880b84c1b981e6fddba673b6f (MD5) Poucos experimentos têm sido conduzidos utilizando o princípio da regeneração óssea guiada nas perfurações ósseas do seio maxilar. Objetivo: Avaliar o processo de reparo ósseo em defeitos cirúrgicos experimentais realizados na parede alveolar do seio maxilar, comunicando-se com a cavidade sinusal utilizando membranas de colágeno e fáscia temporal. Materiais e Métodos: Neste estudo prospectivo, experimental, foram realizados defeitos ósseos em paredes alveolares dos seios maxilares comunicando-se com as cavidades sinusais de quatro macacos pregos (Cebus apella) com membranas de colágeno Gen-derm - Genius Baumer, Pro-tape - Proline e fáscia temporal autóloga e análises histológicas após 180 dias. Resultados: No defeito experimental sem cobertura com métodos de barreira predominou proliferação óssea em dois animais e nos outros dois, tecido conjuntivo fibroso; no grupo em que foi utilizado fáscia temporal predominou tecido conjuntivo fibroso em três animais e proliferação óssea em um animal; nos grupos com membranas de colágeno Gen-derm Protape houve completa proliferação óssea em três animais e predomínio de tecido conjuntivo fibroso em um animal. Conclusões: Os defeitos cirúrgicos experimentais podem ser reparados, tanto por tecido ósseo como por tecido conjuntivo fibroso, e o uso de membranas de colágeno trouxeram benefícios ao processo de reparo ósseo Few studies has been done using guided bone regeneration in maxillary sinus defects. AIM: To assess the bone repair process in surgical defects on the alveolar wall of the monkey maxillary sinus, which communicates with the sinus cavity, by using collagen membranes: Gen-derm - Genius Baumer, Pro-tape - Proline and autologous temporal fascia. MATERIALS AND METHODS: In this prospective and experimental study, orosinusal communications were performed in four tufted capuchin monkeys (Cebus apella) and histologic analysis was carried out 180 days after. RESULTS: In the defects without a cover (control), bone proliferation predominated in two animals and fibrous connective tissue predominated in the other two. In defects repaired with a temporal fascia flap, fibrous connective tissue predominated in three animals and bone proliferation predominated in one. In the defects repaired with Gen-derm or Pro-tape collagen membranes there was complete bone proliferation in three animals and fibrous connective tissue in one. CONCLUSIONS: Surgical defect can be repaired with both bone tissue and fibrous connective tissue in all study groups; collagen membranes was more beneficial in the bone repair process than temporal fascia or absence of a barrier. Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Odontologia de Araçatuba, Aracatuba, Rua José Bonifácio, 1193, Vila Mendonça, CEP 16015-050, SP, Brasil Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Odontologia de Araçatuba, Aracatuba, Rua José Bonifácio, 1193, Vila Mendonça, CEP 16015-050, SP, Brasil

Aim: To describe the early healing processes around the implants installed after elevation of the sinus mucosa applying the lateral access technique without the use of grafting material. Material and methods: Immediately after the elevation of the maxillary sinus Schneiderian membrane by the lateral approach in eight monkeys, implants were installed without the use of grafting material. The healing of the tissue around the implants was evaluated after 4, 10, 20 and 30 days. Ground sections were prepared and analyzed histologically. Results: After 4 days of healing, the formation of coagulum and provisional matrix was documented within the elevated area. At 10-day interval, sprouts of woven bone were in continuity with the parent bone, and partly in contact with the implant surface at the base of the augmented area. While bone-to-implant contact increased after 20 and 30 days, the area underneath the Schneiderian membrane appeared reduced in volume and condensed toward the apex of the implants. The sinus mucosa was to some extent collapsed onto the implant surface and on the newly formed bone. Conclusions: The void initially occupied by the coagulum after sinus membrane elevation shrank substantially during the observation period. A lack of influence of the Schneiderian membrane in bone formation apical to implants was documented in the early phase of healing. To cite this article: Scala A, Botticelli D, Faeda RS, Rangel IG Jr, de Oliveira, JA, Lang NP. Lack of influence of the Schneiderian membrane in forming new bone apical to implants simultaneously installed with sinus floor elevation: an experimental study in monkeys. Clin. Oral Impl. Res. 23, 2012; 175–181. doi: 10.1111/j.1600-0501.2011.02227.x

Aim: To describe the early healing within the void obtained after the elevation of the sinus mucosa and simultaneous implant installation without the use of any grafting material in monkeys. Material and methods: Implants were installed simultaneously with the elevation of the maxillary sinus using the lateral approach in eight monkeys without the use of grafting material. The healing after 4, 10, 20 and 30 days was evaluated in the area distal to the implants. Paraffin sections were prepared and analyzed using qualitative histological methods. Results: The healing process was initiated by the formation of a coagulum and followed by a provisional matrix and woven bone. Subsequently, a parallel-fiber bone replaced woven bone. The dimension of the elevated area shrank during the healing process. Sprouts of woven bone, present to a moderate extent after 4 days, were more numerous after 10 and 20 days. Newly formed bone originated from the sinus walls and septa, while there was no evidence of participation of the Schneiderian membrane in this process. After 30 days, the window access appeared to be closed by a layer of newly formed trabecular bone. Conclusions: The coagulum that filled the void distal to the implant after simultaneous elevation of the sinus floor gave rise to newly formed bone. However, the void occupied by the coagulum shrank substantially. The Schneiderian membrane did not provide a basis for new bone formation in the early phase of healing. To cite this article: Scala A, Botticelli D, Rangel IG Jr, de Oliveira JA, Okamoto R, Lang NP. Early healing after elevation of the maxillary sinus floor applying a lateral access: a histological study in monkeys. Clin. Oral Impl. Res. 21, 2010; 1320–1326. doi: 10.1111/j.1600-0501.2009.01964.x

Osteogenesis imperfecta is a hereditary disorder that affects connective tissue. This disorder may be classified into seven types, according to clinical presentation, radiographic findings, mode of inheritance and histologic features, and into subtypes, depending on the absence or presence of dentinogenesis imperfecta. The most relevant clinical manifestations are represented by bone fragility, growth deficiency, blue sclera, hearing impairment and joint hypermobility. Craniofacial deformities include increased rates of open bite, anterior and/or posterior crossbite, class III occlusal relationship, prominent frontal bossae and short facial height. The aim of the present article is to report a familial case of osteogenesis imperfecta associated with dentinogenesis imperfecta. Clinical manifestations included short stature and the medical history of several bone fractures. Dentinogenesis imperfecta was also present, characterized by brown teeth, low incidence of caries, cracking and loss of enamel on occlusal surfaces caused by attrition. Radiographic findings included obliterated pulp chambers and root channels, although they were open in immature teeth. Similar characteristics were observed in the younger sister. The familiar history disclosed the first occurrence in the father, as he related four femoral fractures and had also manifested dentinogenesis imperfecta.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:44:19Z No. of bitstreams: 1 S1808-86942009000300007.pdf: 1253459 bytes, checksum: 5b01351c327749aa396bb8f311cdf3a0 (MD5) Made available in DSpace on 2013-08-22T18:44:19Z (GMT). No. of bitstreams: 1 S1808-86942009000300007.pdf: 1253459 bytes, checksum: 5b01351c327749aa396bb8f311cdf3a0 (MD5) Previous issue date: 2009-06-01 Made available in DSpace on 2013-09-30T18:29:18Z (GMT). No. of bitstreams: 2 S1808-86942009000300007.pdf: 1253459 bytes, checksum: 5b01351c327749aa396bb8f311cdf3a0 (MD5) S1808-86942009000300007.pdf.txt: 21343 bytes, checksum: c3221a915d2b6fbae1d4d4b278fe085e (MD5) Previous issue date: 2009-06-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:43:05Z No. of bitstreams: 2 S1808-86942009000300007.pdf: 1253459 bytes, checksum: 5b01351c327749aa396bb8f311cdf3a0 (MD5) S1808-86942009000300007.pdf.txt: 21343 bytes, checksum: c3221a915d2b6fbae1d4d4b278fe085e (MD5) Made available in DSpace on 2014-05-20T13:43:05Z (GMT). No. of bitstreams: 2 S1808-86942009000300007.pdf: 1253459 bytes, checksum: 5b01351c327749aa396bb8f311cdf3a0 (MD5) S1808-86942009000300007.pdf.txt: 21343 bytes, checksum: c3221a915d2b6fbae1d4d4b278fe085e (MD5) Previous issue date: 2009-06-01 Lesões do tecido ósseo podem ser causadas por fatores congênitos e adquiridos e resultar em deformidade nasal com repercussão estética e funcional. O tratamento cirúrgico desses casos requer reconstruções complexas e frequentemente o uso de biomateriais. O poliuretano derivado do óleo da mamona apresenta uma fórmula com aspectos favoráveis de processabilidade, flexibilidade de formulação, ausência de emissão de vapores tóxicos e baixo custo. Entretanto, a despeito dos resultados favoráveis, estudos referentes ao uso do polímero de mamona, avaliando a reação tecidual no dorso nasal, ainda não foram realizados. OBJETIVO: O objetivo deste estudo consiste em avaliar histologicamente a biocompatibilidade do implante do polímero de mamona no dorso nasal. FORMA DE ESTUDO: Experimental. MATERIAL E MÉTODO: Foram utilizados quatro macacos-pregos da espécie Cebus apella. Um defeito ósseo foi realizado no osso nasal em todos os animais e colocado um implante de polímero de mamona. A eutanásia foi realizada com 270 dias de pós-operatório, e as amostras foram submetidas a estudo histológico. RESULTADOS: Na análise histológica não foi observada a presença de granuloma de corpo estranho ou células fagocitárias. Progressiva formação óssea e maturação foram observadas. CONCLUSÃO: Os resultados macroscópicos e microscópicos mostraram que o implante de polímero de mamona foi biocompatível. Bone tissue lesions can be caused by congenital and acquired factors, and result in nasal deformities with cosmetic and functional repercussion. Surgical treatment in these cases frequently requires complex reconstructions and the use of biomaterials. The polyurethane derived from castor beans (Ricinus communis) has a favorable formulation in terms of ease of processing, flexibility, no emission of toxic vapors and low cost. Nonetheless, despite favorable results, studies about the use of castor beam polymer (Ricinus communis) assessing tissue reaction on the nasal dorsum are still missing in the literature. AIM: the goal of the present investigation is to histologically assess the Ricinus communis polymer implant biocompatibility with the nasal dorsum. STUDY DESING: experimental. MATERIALS and METHODS: we used four Cebus appela monkeys, in which we created a nasal dorsal defect in all the animals and there we placed the aforementioned implant. The animals were sacrificed 270 days after surgery and the samples were submitted to histological study. RESULTS: in the histology analysis we did not observe the presence of foreign body granulomas or phagocytic cells. We also observed a progressive bone formation and maturation. CONCLUSION: macroscopic and microscopic results showed that the castor oil polymer implant was biocompatible. Federal University of São Paulo Universidade Federal do Rio Grande do Sul (UFRGS) Hospital de Clinicas de Porto Alegre Santa Casa de São Paulo Faculdade de Ciências Médicas UNESP Faculdade de Odontologia de Araraquara Department of Morphology UNESP Faculdade de Odontologia do Campus de Araçatuba Departamento de Ciências Básicas UNESP Faculdade de Odontologia de Araraquara UNESP Faculdade de Odontologia de Araraquara Department of Morphology UNESP Faculdade de Odontologia do Campus de Araçatuba Departamento de Ciências Básicas UNESP Faculdade de Odontologia de Araraquara

The purpose of this experimental study was to evaluate the collagen fiber distribution histologically after phenytoin, cyclosporin, or nifedipine therapy and to correlate it with collagen I and matrix metalloproteinase (MMP)-1 and -2 gene expression levels.Gingival samples from the canine area were obtained from 12 male monkeys (Cebus apella). The mesial part of each sample was assessed by reverse transcription-polymerase chain reaction, whereas the distal part was processed histologically for picrosirius red and hematoxylin and eosin stainings, as well as for collagen IV immunostaining. One week after the first biopsy, the animals were assigned to three groups that received daily oral dosages of cyclosporin, phenytoin, or nifedipine for 120 days. Additional gingival samples were obtained on days 52 and 120 of treatment from two animals from each group on the opposite sides from the first biopsies.Picrosirius red staining showed a predominance of mature collagen fibers in the control group. Conversely, there was an enlargement of areas occupied by immature collagen fibers in all groups at days 52 and 120, which was not uniform over each section. There was a general trend to lower levels of MMP-1 gene expression on day 52 and increased levels on day 120. Phenytoin led to increased levels of MMP-2 and collagen I gene expression on day 120, whereas the opposite was observed in the nifedipine group.Cyclosporin, phenytoin, and nifedipine led to phased and drug-related gene expression patterns, resulting in impaired collagen metabolism, despite the lack of prominent clinical signs.

This study evaluated the antifungal activity ofPersea americanaextract onCandida albicansbiofilm and its cytotoxicity in macrophage culture (RAW 264.7). To determine the minimum inhibitory concentration (MIC), microdilution in broth (CLSI M27-S4 protocol) was performed. Thereafter, the concentrations of 12.5, 25, 50, 100, and 200 mg/mL(n=10)with 5 min exposure were analyzed on mature biofilm in microplate wells for 48 h. Saline was used as control(n=10). After treatment, biofilm cells were scraped off and dilutions were plated on Sabouraud dextrose agar. After incubation (37°C/48 h), the values of colony forming units per milliliter (CFU/mL) were converted to log10and analyzed (ANOVA and Tukey test, 5%). The cytotoxicity of theP. americanaextract was evaluated on macrophages by MTT assay. The MIC of the extract was 6.25 mg/mL and with 12.5 mg/mL there was elimination of 100% of planktonic cultures. Regarding the biofilms, a significant reduction(P<0.001)of the biofilm at concentrations of 50 (0.580±0.209log10), 100 (0.998±0.508log10), and 200 mg/mL (1.093±0.462log10) was observed. The concentrations of 200 and 100 mg/mL were cytotoxic for macrophages, while the concentrations of 50, 25, and 12.5 mg/mL showed viability higher than 55%.

Made available in DSpace on 2019-09-12T16:53:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2015 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Objective: The role of matrix metalloproteinases (MMPs) in tissue degradation has become evident in many diseases and great interest therefore exists in the pharmacological control of the activity of these enzymes. This study evaluated the effect of caffeic acid phenethyl ester (CAPE) on the production of MMPs and their inhibitor (TIMP) in monocytes activated by lipopolysaccharide (LPS). Design: The human monocytic cell line (THP-1) was treated with non-cytotoxic concentrations of CAPE (10 and 60 mu M) combined with 1 mu g/mL of LPS. The gene expression of MMP-1, MMP-9 and TIMP-1 was evaluated by quantitative real-time polymerase chain reaction. The protein secretion into the culture medium was assessed via enzyme-linked immunosorbent assay and the gelatinolytic activity of MMP-9 by zymography. Results: CAPE, especially at the highest concentration, down-regulated MMP-1 and MMP-9 gene expression but up-regulated the gene expression of TIMP-1. Furthermore, CAPE reduced the secreted protein level of MMP-1 and MMP-9 as well as the gelatinolytic activity of MMP-9. Conclusion: CAPE was able to inhibit the gene expression, production and the activity of MMPs induced by LPS and also increased the gene expression of TIMP-1. The present observations suggest that CAPE exerted a positive effect on the regulatory mechanism between MMPs and TIMP, which is important for the control of different diseases. (C) 2015 Elsevier Ltd. All rights reserved. [Figueiredo Vilela, Polyana das Gracas; de Oliveira, Jonatas Rafael; de Barros, Patricia Pimentel; Cardoso Jorge, Antonio Olavo] Univ Estadual Paulista, Lab Microbiol & Immunol, Dept Biosci & Oral Diag, Inst Sci & Technol,UNESP, Sao Jose Dos Campos, SP, Brazil [Pereira Leao, Mariella Vieira] Universidade de Taubaté (Unitau), Biosci Basic Inst [de Oliveira, Luciane Dias] Univ Estadual Paulista, Lab Biochem & Pharmacol, Dept Biosci & Oral Diag, Inst Sci & Technol,UNESP, Sao Jose Dos Campos, SP, Brazil

We studied the distribution of NADPH-diaphorase (NADPH-d) activity in the prefrontal cortex of normal adult Cebus apella monkeys using NADPH-d histochemical protocols. The following regions were studied: granular areas 46 and 12, dysgranular areas 9 and 13, and agranular areas 32 and Oap. NADPH-d-positive neurons were divided into two distinct types, both non-pyramidal. Type I neurons had a large soma diameter (17.24 +/- 1.73 microm) and were densely stained. More than 90% of these neurons were located in the subcortical white matter and infragranular layers. The remaining type I neurons were distributed in the supragranular layers. Type II neurons had a small, round or oval soma (9.83 +/- 1.03 microm), and their staining pattern varied markedly. Type II neurons were distributed throughout the cortex, with their greatest numerical density being observed in layers II and III. In granular areas, the number of type II neurons was up to 20 times that of type I neurons, but this proportion was smaller in agranular areas. Areal density of type II neurons was maximum in the supragranular layers of granular areas and minimum in agranular areas. Statistical analysis revealed that these areal differences were significant when comparing some specific areas. In conclusion, our results indicate a predominance of NADPH-d-positive cells in supragranular layers of granular areas in the Cebus prefrontal cortex. These findings support previous observations on the role of type II neurons as a new cortical nitric oxide source in supragranular cortical layers in primates, and their potential contribution to cortical neuronal activation in advanced mammals.