Tratamento de feridas excisionais de coelhos com extrato de barbatimão associado a células mononucleares autólogas da medula óssea / Treatment of excisional wound in rabbits with barbatiman extracts associated with autologous bone marrow mononuclear cells

Objetivou-se com este estudo avaliar o processo de cicatrização de feridas de coelhos tratadas com extrato de barbatimão (S. adstringens) associado a células mononucleares autólogas da medula óssea (CMMO). Utilizaram-se 20 coelhos, distribuídos em quatro grupos: B, extrato de barbatimão; CB, CMMO com extrato de barbatimão; CS, CMMO com solução fisiológica; S, solução fisiológica. Foi avaliada a presença de crosta, hiperemia, secreção, hemorragia, reepitelização, área da ferida e tempo de cicatrização. No terceiro, sétimo, 14º e 21º dias pós-operatório, realizou-se a biópsia das feridas e avaliaram-se os indicadores dos processos de inflamação e de reparo, com destaque para o colágeno, na coloração picrosírius, bem como de proliferação celular, na coloração AgNOR. Houve maior deposição de fibras colágenas nos grupos B e CB (P=0,00003) e formação de crostas mais espessas no sétimo dia, com fibras colágenas mais organizadas no 21º dia. Conclui-se que o barbatimão estimula a produção de fibras colágenas e promove a formação de crostas mais espessas sobre a ferida na fase inicial da cicatrização e, na fase de remodelação, favorece a orientação das fibras colágenas. Além disso, a associação desse fitoterápico com CMMO não estimula a cicatrização de feridas.(AU)

This study aimed to evaluate the healing process of wounds of rabbits in response to treatment with barbatiman extract (S. adstringens) associated with autologous mononuclear bone marrow cells (BM-MNC). We used 20 rabbits, divided into four groups: B, 10% barbatiman extract with 9.48% of total tannins; CB, BM-MNC with barbatiman extract; CS, BM-MNC with NaCl 0.9% solution; S, NaCl 0.9% solution. Clinical evaluation was performed by observing the presence of crust, redness, discharge, bleeding, re-epithelialization, the wound area and healing time in days. In the third, seventh, 14th and 21st postoperative days wounds were biopsied for microscopic evaluation of inflammation and repair process indicators, especially collagen, in picrosirius staining, and cell proliferation, in AgNOR staining. There was a greater deposition of collagen fibers in groups B and CB (p=0.00003) on the seventh day and formation of thicker crusts, and more organized collagen fibers on the 21st day in these groups. In conclusion, in the initial phase of healing, barbatiman extract stimulates the production of collagen fibers and promotes the formation of more exuberant crusts on the wounds and remodeling phase favors the orientation of collagen fibers, but when combined with BMMC does not stimulate wound healing in rabbits.(AU)

Avaliação da atividade proliferativa celular das lesões bucais causadas pelo uso de próteses totais através da impregnação tecidual pela prata / Proliferative cellular activity evaluation of lesions caused by dentures through tissue silver impregnation

Introdução: As próteses totais visam conservar a função do sistema estomatognático do paciente totalmente edêntulo. Porém, na mucosa bucal podem aparecer manifestações cuja principal causa são as próteses totais mal adaptadas. Objetivo: o presente estudo objetiva investigar a proliferação tecidual das lesões causadas por próteses totais removíveis através do método de impregnação pela prata (AgNOR), com isso facilitando o tratamento e a determinação do prognóstico das lesões a serem estudadas. Metodologia: foram selecionados todos os casos das lesões bucais mais comumente associadas à utilização de próteses totais registradas no Serviço de Diagnóstico Histopatológico do ICB­UPF nos anos de 2012 e 2013, tendo sido encontrados 5 casos de granuloma piogênico, 5 casos de hiperplasia de fundo de sulco, 5 casos de fibroma de irritação e 2 casos de fibroma ossificante periférico. Os cortes histopatológicos das lesões foram impregnados pela prata (método AgNOR), tendo sido obtido, com auxílio do programa Image Tool®, o número de NORs de 100 células de cada caso, resultando numa média de NORs em cada grupo de lesões. Resultados: os resultados obtidos foram tabulados em planilha eletrônica e a comparação do número médio de NORs de cada grupo foi realizado por meio do teste estatístico ANOVA, 5% de significância. Resultados: o grupo das hiperplasias de fundo de sulco mostrou média de 2,41 NORs por núcleo, o grupo dos granulo mas piogênicos mostrou 2,44, o fibroma de irritação 2,22, e o fibroma ossificante periférico mostrou média de 1,89 NORs por núcleo celular, diferindo estatisticamente esta lesão das anteriormente mencionadas (p = 0,002). Conclusão: o fibroma ossificante periférico mostrou ser a lesão causada por prótese total removível com a menor atividade proliferativa celular. Tal estudo precisa ser complementado por futuros estudos clínicos.

Introduction: the total dentures are aimed at preserving the function of the stomatognathic system of the fully edentulous patient. However, in the oral mucosa may appear manifestations whose main cause are the totally unsuitable dentures. Objective: this study aims to investigate the proliferation of tissue lesions caused by removable dentures by impregnation method for silver (AgNOR), thereby facilitating the treatment and determining the prognosis of the lesions to be studied. Methodology: we selected all cases of oral lesions most commonly associated with the use of dentures recorded in Histopathological Diagnostic Service ICB-UPF in the years 2012 and 2013, having been found 5 cases of pyogenic granuloma, 5 cases of hyperplasia, 5 cases of irritation fibroma and 2 cases of peripheral ossifying fibroma. Histopathological lesions cuts were impregnated by silver (AgNOR method), having been obtained with the aid of the program Image Tool®, the number of NOR cells 100 in each case, resulting in an average NORs in every group of lesions. Results: the results were tabulated in a spreadsheet and comparing the average number of NORs of each group was conducted through ANOVA, 5% significance level. Results: The group of hyperplasias showed average of 2.41 NORs per nucleus, the group of pyogenic granulomas showed 2.44, the irritation fibroma 2.22, and peripheral ossifying fibroma showed average of 1.89 NORs for cell nucleus, differing significantly from that of the aforementioned lesions (p = 0.002). Conclusion: the peripheral ossifying fibroma proved the injury caused by removable dentures with lower cell proliferative activity. This study needs to be complemented by future clinical studies.

Avaliação da técnica de coloração AgNOR em testículos de ovinos / Evaluation of the AgNOR staining method in ovine testicles

A coloração pela prata das regiões organizadoras de nucléolos (NORs) é caracterizada por marcar proteínas ligadas ao ácido ribonucleico ribossômico, avaliando a proliferação em células normais ou neoplásicas. Objetivou-se estudar, em testículos de ovinos obtidos em matadouro, a validade do uso da técnica de coloração pela prata (AgNOR) na identificação das regiões organizadoras de nucléolo (NORs) em células saudáveis da linhagem espermatogênica. Utilizaram-se 43 pares de testículos de ovinos mestiços entre seis e 10 meses de idade. Testes de Wilcoxon e Spearman foram empregados, com nível de 5%. As médias das NORs nas células das gônadas direita e esquerda foram, respectivamente: espermatogônia (8,77±1,14 e 9,04±0,96), espermatócitos (4,99±2,00 e 6,20±2,07; P<0,05), Leydig (8,05±2,82 e 7,89±2,29) e Sertoli (8,07±1,88 e 7,61±2,16; P<0,05). Houve correlação (P<0,05) entre os lados para o número de NORs: espermatócitos x Leydig (0,49); espermatócitos x Sertoli (0,49) e Leydig x Sertoli (0,96). Conclui-se ser válido o emprego da técnica AgNOR para avaliar o potencial proliferativo das células saudáveis em testículos de ovinos com prática execução e baixo custo.(AU)

The silver staining technique for AgNOR nucleolar organizer regions (NORs) is characterized by marking proteins linked to the ribosomal ribonucleic acid, evaluating cell proliferation. The aim was to study the validity of the AgNOR staining technique in the testicular cell proliferation in crossbred ovine. Forty-three pairs of ovine testicles between 6 and 10 months old were collected. Wilcoxon and Spearman tests were used with a significance level of 5%. The mean NORs count in cells of the right and left gonads were respectively: spermatogonia (8.77±1.14 and 9.04±0.96), spermatocytes (4.99±2.00 and 6.20±2.07, P<0.05), Leydig (8.05±2.82 and 7.89±2.29) and Sertoli cells (8.07±1.88 and 7.61±2.16; P<0.05). There was a correlation between the mean values for the right and left sides for the number of NORs (P<0.05) between Leydig x spermatocytes (0.49); spermatocytes x Sertoli (0.49) and Sertoli x Leydig (0.96). The study demonstrates that the AgNOR staining technique is indicated to evaluate the cell proliferative potential in ovine testis with practical implementation and low cost.(AU)

Novel biological activity of ameloblastin in enamel matrix derivative

Objective Enamel matrix derivative (EMD) is used clinically to promote periodontal tissue regeneration. However, the effects of EMD on gingival epithelial cells during regeneration of periodontal tissues are unclear. In this in vitro study, we purified ameloblastin from EMD and investigated its biological effects on epithelial cells. Material and Methods Bioactive fractions were purified from EMD by reversed-phase high-performance liquid chromatography using hydrophobic support with a C18 column. The mouse gingival epithelial cell line GE-1 and human oral squamous cell carcinoma line SCC-25 were treated with purified EMD fraction, and cell survival was assessed with a WST-1 assay. To identify the proteins in bioactive fractions of EMD, we used proteome analysis with two-dimensional gel electrophoresis followed by identification with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Results Purified fractions from EMD suppressed proliferation of GE-1 and SCC-25. LC-MS/MS revealed that ameloblastin in EMD is the component responsible for inhibiting epithelial cell proliferation. The inhibitory effect of ameloblastin on the proliferation of GE-1 and SCC-25 was confirmed using recombinant protein. Conclusion The inhibitory effects of EMD on epithelial cell proliferation are caused by the biological activities of ameloblastin, which suggests that ameloblastin is involved in regulating epithelial downgrowth in periodontal tissues. .

Adaptación de la técnica de impregnación argéntica de Llombart para la demostración de fibras nerviosas, en cualquier tejido en cortes por parafina / Adaptation of Llombart's silver impregnation technique for demonstration of nerve fibers in any tissue in paraffin sections

En la investigación biológica sigue siendo necesaria la demostración de la inervación periférica en numerosos tejidos y órganos. El objetivo de este trabajo fue rescatar y modernizar uno de los métodos más constantes que hemos probado para demostrar la inervación periférica. La técnica de Llombart para fibras nerviosas se adaptó en cortes por parafina de 7 µm en diferentes tejidos animales. La impregnación argéntica se hizo por goteo en cámara húmeda. Se demostraron en forma constante, precisa y seriada terminaciones nerviosas y corpúsculos sensoriales, neuronas y fibras nerviosas periféricas. A pesar de la alta especificidad para fibras nerviosas, la técnica no compromete el panorama tisular por lo que da bellas imágenes de conjunto. Sin ser una técnica para argentafinidad, demuestra claramente dos tipos de células argentafines en las glándulas adrenales. La adición de los reactivos metálicos en gotas y en cámara húmeda, ofrece una variante sumamente económica.

In Biological research is still necessary for the demonstration of the peripheral innervation in numerous tissues and organs. The aim of this study was to rescue and modernize one of the most consistent methods that we have tried to demonstrate peripheral innervation. Llombart's technique for nerve fibers was adapted by paraffin cuts of 7 µm in different animal tissue. The silver impregnation was done by dripping in a moist chamber. It was demonstrated in a constant, precise and serial form, nerve terminations, and sensorial corpuscles, neurons, and peripheral nerve fibers. Despite being highly specific to nerve fibers, the technique does not sacrifice tissue panorama so it gives beautiful images set. Without being a technique to argentaffin structures, it clearly shows two types of argentaffin cells in the adrenal glands. The addition of the metal reactive in droplets and in a humid chamber provides a very economical variant.

Detection of leptospires from infected urine and tissue samples in vitro by modified Fontana silver stain / Detecção de leptospiras na urina e nos tecidos infectados in vitro por impregnação com prata Fontana modificada

INTRODUCTION: The microbiological diagnosis of leptospirosis comprises bacteriological and serological methods. The former ones allow the direct detection of leptospires and are considered presumptive with the exception of culture. Therefore, they constitute invaluable tools for rapid diagnosis, mainly in samples from deceased subjects. OBJECTIVE: To evaluate a modified Fontana silver staining method in experimentally infected samples. MATERIAL AND METHODS: Human and animal (hamster) urine samples were experimentally infected with different strains of Leptospira interrogans sensu lato. Liquid culture medium, leptospira cultures, experimentally infected and non-infected human urine samples, clarified and non-clarified imprints, and clarified and non-clarified suspension smears from tissues of experimentally infected and non-infected hamsters were applied for the ass essment of silver staining. The analytical sensitivity of the assay was compared with dark field microscopy and culture. Other bacterial and fungi species were also used. RESULTS: The modified Fontana silver staining allowed the accurate observation of the well-defined leptospire helical structure. On leptospire cultures from infected human samples, we could observe until (1-10) × 10³ leptospires/ml, higher sensitivity in comparison with direct dark field microscopy and lower in comparison with culture. The best results in tissues were obtained on clarified imprints and non-clarified suspension smears. Morphological and stainable structures compatible with leptospires were not observed in the samples without them. CONCLUSION: This procedure allowed differentiating the characteristic morphology of leptospires. As its application suggests, it consists of a simple and easily conducted procedure with stable reagents.

INTRODUÇÃO: O diagnóstico microbiológico da leptospirose inclui métodos bacteriológicos e sorológicos; os primeiros permitem a detecção direta de leptospiras e, à exceção do cultivo, são considerados como presuntivos, mas constituem ferramentas valiosas para o diagnóstico rápido, principalmente nos falecidos. OBJETIVO: Avaliar a coloração de prata Fontana modificada em amostras experimentalmente infectadas. MATERIAL E MÉTODOS: Urinas humanas e hamsters foram infectados experimentalmente com diferentes cepas de Leptospira interrogans sensu lato. Para a avaliação, foi utilizado meio de cultura líquido, culturas de leptospiras, urinas humanas experimentalmente infectadas e não infectadas, impressões clarificadas e não clarificadas e esfregaços de suspensões clarificadas e não clarificadas a partir de tecidos de hamsters infectados e não infectados para o experimento. A sensibilidade analítica do ensaio foi comparada com a microscopia de campo escuro e de cultura. Outras espécies de bactérias e fungos também foram utilizadas. RESULTADOS: A coloração de prata de Fontana modificada permitiu observar claramente e bem definida a estrutura helicoidal das leptospiras. Nas culturas destas e de urinas humanas infectadas, pôde-se observar até (1-10) × 10³ leptospiras/ml, sensibilidade superior à da microscopia de campo escuro e inferior à da cultura. Os melhores resultados nos tecidos foram obtidos em impressões clarificadas e a partir de esfregaços de suspensões não clarificadas. Estruturas morfológicas e tingidas compatíveis com leptospiras não foram observadas nas amostras livres destas. CONCLUSÃO: Esse procedimento permitiu diferenciar a morfologia característica das leptospiras. É um procedimento simples, fácil de realizar e com reagentes estáveis pelo o que a sua aplicação é sugerida.

Tinción de Gram de tejido: alcances y limitaciones / Gram stain of tissue biopsy: scope and pitfalls

La identificación de microorganismos en tejido es esencial para reconocer un proceso infeccioso. Inicialmente, mediante la coloración de hematoxilina-eosina, se puede identificar el patrón de inflamación asociado y luego, a través de tinciones basadas en plata y la tinción de Gramde tejido se visualizan los microorganismos. La tinción de Gram no solo sirve para bacterias, sino también para algunos hongos y parásitos; no obstante, esta técnica tiene algunos inconvenientes, como la contaminación con otros microorganismos y la imposibilidad de visualizar algunas bacterias, entre ellas Legionella pneumophila, Leptospira spp y Bartonella spp. En este artículode revisión se describirán los fundamentos del Gram de tejido, su contribución en el diagnóstico de infecciones como herramienta adicional para el reconocimiento de microorganismos, y sus limitaciones.

The identification of microorganisms in tissue is pivotal to recognize infectious processes.At first, the hematoxylin-eosin stain is used to identify the pattern of inflammation associated; after that, microorganisms are seen through Gram or silver stains. Gram Stain of tissue biopsy not only stains bacteria, but also a number of fungus and parasites. However, this technique has some disadvantages, such as contamination with other microorganisms, and lack of stain of some bacteria, including Legionella pneumophila, Leptospira spp and Bartonella spp. This review articleaims to describe the fundamentals of Gram stain of tissue biopsy and its assistance in infectious diagnosis as an additional tool for recognition of microorganisms, as well as its limitations.

Histologic grading and nucleolar organizer regions in oral squamous cell carcinomas

OBJECTIVE: The purposes of this study were to histologically assess different types of oral squamous cell carcinoma and the silver-binding nucleolar organizer region (AgNOR) morphology in neoplastic cells, as well as to quantify the number of AgNORs in each type of carcinoma in order to relate AgNOR count and histologic grading. MATERIAL AND METHODS: Twenty-eight cases of oral squamous cell carcinoma were divided into 4 groups, namely well-differentiated, moderately differentiated, poorly differentiated, and undifferentiated. For NOR study, 3-µm-thick sections were stained with 50 percent aqueous silver nitrate solution. The predominant microscopic pattern of NORs was determined. Quantitative analyses of NORs were obtained of all cells present on each histological field using a 0.025 mm² eyepiece graticule. Different histological fields were analyzed until the total number of NORs was 120 cells for each tumor. Kruskall-Wallis test was applied to compare the groups of sample data at a significance level of p=0.05. RESULTS: The mean number of AgNORs per nucleus was 3.20 for the well-differentiated group, 5.33 for the moderately differentiated one, 8.27 for the poorly differentiated one, and 10.08 for the undifferentiated one. AgNOR count was significantly different (p<0.05) among all of the studied groups. CONCLUSION: AgNOR staining technique seems to be a useful diagnostic tool since differences in AgNOR numeric values can be identified in the different types of oral squamous cell carcinoma. This technique is easy to handle and inexpensive, thus justifying its large use in histopathology.

A metallic impregnation technique adapted to study the honeybee Apis mellifera L. brain

In order to visualize the distribution pattern of the neuronal bodies and neurofibrils in the honeybee brain, we adapted a metallic impregnation technique first described for vertebrate nervous system by Ramón y Cajal. The honeybee brain constitution plays a key role in the development of learning and memory capacities. The general characteristics observed in the honeybee brain, stained by metallic impregnation, revealed its anatomical and morphological constitution in agreement with studies of other insect brains using different techniques. Metallic impregnation evidenced the optic lobe neuropils, the ocelli fiber cells, the neuron extensions of the calyces, and the axon bundles that involve the antennal glomeruli, as well as the neuron extensions in the alpha lobe. We also observed that the antennal glomeruli were mainly formed by fibers. The optical lobes were impregnated distinctly in the monopolar neuron bodies and in the fibers. In the mushroom bodies, we observed the lip, collar and calyx basal areas. Based on our results, the metallic impregnation technique is effective to visualize neuronal bodies and neurofibrils; moreover, is simpler and faster than other techniques, offering new insights for the investigation of the invertebrate nervous system.

Microleakage in conservative cavities varying the preparation method and surface treatment

OBJECTIVE: To assess microleakage in conservative class V cavities prepared with aluminum-oxide air abrasion or turbine and restored with self-etching or etch-and-rinse adhesive systems. Materials and Methods: Forty premolars were randomly assigned to 4 groups (I and II: air abrasion; III and IV: turbine) and class V cavities were prepared on the buccal surfaces. Conditioning approaches were: groups I/III - 37 percent phosphoric acid; groups II/IV - self-priming etchant (Tyrian-SPE). Cavities were restored with One Step Plus/Filtek Z250. After finishing, specimens were thermocycled, immersed in 50 percent silver nitrate, and serially sectioned. Microleakage at the occlusal and cervical interfaces was measured in mm and calculated by a software. Data were subjected to ANOVA and Tukey's test (α=0.05). RESULTS: Marginal seal provided by air abrasion was similar to high-speed handpiece, except for group I. There was SIGNIFICANT difference between enamel and dentin/cementum margins for to group I and II: air abrasion. The etch-and-rinse adhesive system promoted a better marginal seal. At enamel and dentin/cementum margins, the highest microleakage values were found in cavities treated with the self-etching adhesive system. At dentin/cementum margins, high-speed handpiece preparations associated with etch-and-rinse system provided the least dye penetration. CONCLUSION: Marginal seal of cavities prepared with aluminum-oxide air abrasion was different from that of conventionally prepared cavities, and the etch-and-rinse system promoted higher marginal seal at both enamel and dentin margins.

Effect of finasteride on serum prostate-specific antigen (PSA) and on prostate of hamster Mesocricetus auratus (hMa) / Efeito da finasterida no antígeno prostático específico (PSA) sérico e na próstata do hamster Mesocricetus auratus (hMa)

PURPOSE: Evaluate the effects of finasteride on the serum PSA and on the prostate of hamster-Mesocricetus auratus(hMa). METHODS: Twenty hMa male adults were split in groups control and experimental (n=10). Animals of the experimental group received 7.14ng/mL of finasteride, subcutaneously (SC) on the back three times per week, during 90 days. The finasteride dose was equivalent to 5.0mg administered to a 70kg man. At the end of the experiment the mean age for the animals in the control group was 15.2±1.13months and for the experimental group was 17.7±0.67 months. There was a statistically significant difference between mean ages of both groups (t value=5.98; p=0.001). The animals of the control group weighted 129.0±18.8g and the experimental group weighted 145.0±15.5g, t=1.88 e p=0.0514. The serum PSA was assessed through ELISA method. Prostates of those animals were collected and processed to histology and morphometry: the diameter of the acinous glands and the acinous epithelium, apoptosis, AgNORs and cellularity were assessed in both groups. RESULTS: Serum PSA decreased in the experimental group, 0.003ng/mL versus 0.763ng/mL, H= 7.982 e p= 0.0047. Decrease in the acinous area occurred in animals that received finasteride, 238.000±24.600 μm² versus 398.600±55.320 μm²; t= 2.653; p= 0.0122. A remarkable decrease in the area of the acinous epithelium occurred in the animals that received finasteride, 111.900±12.820 μm² versus 160.400±18.430 μm² t= 2.162; p= 0.0361. AgNORs were less expressed in finasteride treated animals, 2.846±0.877 versus 3.68 ±1.07 argyrophilic clusters for μm², p= < 0.0001. Apoptosis was more intense in the experimental group, 53.62±1.389 than in controls, 14.76 ± 2.137, p= 0.0408. However, there was no statistical difference in the cellularity between both groups, 74.75±5.5 cells, in controls versus 65.07±13.24, in treated animals, p=0.5105. CONCLUSIONS: Use of finasteride decreased serum ...

OBJETIVO: Avaliar o efeito da finasterida no PSA sérico e na próstata do hamster-Mesocricetus auratus (hMa). MÉTODOS: 20 hMa adultos machos foram divididos em grupos de 10 animais. No experimento foram administrados 7,14 ng/mL de finasterida, subcutâneo (SC), no dorso, três vezes por semana, por 90 dias, dose equivalente a 5,0 mg usada em homem de 70Kg. Ao final da pesquisa, grupo experimento apresentou idade média de 17,7 ± 0,67 meses. O grupo controle apresentou idade média de 15,2 ± 1,13 meses. O valor de t na comparação das médias das idades entre os dois grupos foi de 5,98 e p=0.0001. Os animais-controle pesaram em média 129,0 ± 18,8g e o experimento 145,0 ± 15,5g; t=1,88 e p=0,0514. Na microscopia óptica de luz e estudo morfométrico: avaliaram-se o diâmetro dos ácinos e epitélio acinar prostáticos, a apoptose, a expressão AgNORs e a celularidade. RESULTADOS: O grupo-experimento apresentou média de PSA de 0,003 ng/mL e o grupo-controle de 0,763 ng/mL, H=7,982 e p=0,0047. A área dos ácinos do grupo-experimento foi de 238,000±24,600 μm² versus 398,600±55,320 μm²; t= 2,653; p= 0,0122. A área do epitélio acinar no grupo-experimento foi de 111,900±12,820 μm² versus 160,400±18,430 μm² t= 2,162; p= 0,0361. A expressão de AgNORs foi menor no grupo-experimento 2,846±0,877 versus 3,68 ±1,07 grumos argilófilos por μm², p= < 0,0001. A apoptose foi mais freqüente no grupo-experimento, 53,62±1,389 versus controle, 14,76 ± 2,137, p= 0,0408. Não houve diferença na celularidade entre os grupos de animais, 74,75±5,5 células no grupo-controle versus 65,07±13,24, no grupo-experimento, p= 0,5105. CONCLUSÕES: A finasterida diminuiu o PSA sérico, a área do lúmen, o epitélio acinar, a expressão de AgNORs e promoveu a apoptose nos ácinos da próstata dos hamsteres experimento e não houve diferença na celularidade acinar entre os animais estudados.

Estudo comparativo da atividade proliferativa celular do fibroma ossificante periférico e da lesão de células gigantes periférica / Comparative study of the proliferative activity of the peripheral giant cells lesion and of peripheral ossifying fibroma

Foi realizado um estudo clínico retrospectivo descritivo e histoquímico de casos do fibroma ossificante periférico (FOP) e da lesão de células gigantes periférica (LCGP), coletados aleatoriamente no arquivo do Serviço de Diagnóstico Histopatológico da Universidade de Passo Fundo. O objetivo do trabalho foi comparar as atividades proliferativas celulares dessas lesões, visando o estabelecimento de uma conduta terapêutica adequada para cada uma das enfermidades. Os dados referentes ao sexo, idade, raça, localização da lesão e ocorrência de recidiva foram analisados e, o número de regiões organizadoras nucleolares (NORs) por núcleo de células ovóides foi determinado por meio do método de impregnação pela prata (Ag-NOR). Os resultados obtidos para o FOP foram de prevalência da lesão em mulheres (70%) da raça branca (60%) com média de idade de 28,11 anos, maior ocorrência da lesão na maxila anterior e índice de recidiva de 30%. O número médio de NORs foi de 1,72 para cada núcleo. Já para a LCGP, o sexo feminino também atingiu 70%, porém a média de idade foi de 42,90 anos e 100% dos pacientes pertenciam à raça branca. A ocorrência da lesão foi maior na região mandibular anterior e apresentou um índice de 20% de recidiva. Nesta lesão, o número médio de NORs foi de 1,93 por núcleo. Contudo, nos testes de correlação, nenhuma das características clínicas apresentou associação com o número médio de NORs por núcleo e a comparação desse número médio por núcleo nas LCGPs e nos FOPs não mostrou diferença significativa entre os grupos.

A retrospective clinical descritive and histochemical study envolving cases of periferic bone fibroma and of periferic giant cell lesion, randomly collected from the archives of Histophatologic Diagnostic Service of University of Passo Fundo was made. The objective of this work was to compare the proliferating cellular activities of these lesions, aiming the establishment of an adequate therapeutic conduct to each one of the disorders. The datas referring to the gender, age, race, position of the lesion and recurrence occurrence were analized and considered and the NORs’ numbers by egg-shaped cells’ nucleus were analized through the impregnation of silver method (Ag-NOR), getting as a result of periferic bone fibroma, 70% women, with an age average of 28.11 years old, where 60% of the pacients were white, and with the lesion’s position more often being on the front upper jaw, with the recurrence’s index up to 30%. The NORs’ average number was of 1.72 to each nucleus. On the periphery giant cell lesion, the female gender had also achieved 70%, however the age’s average was of 42.90 and 100% were white. The occurrence of the lesion was higher on the front inferior jaw and had showed a recurrence’s index of 20%. The NORs’ average of this lesion was 1.93 to each nucleus. Nevertheless, on the correlation’s tests none of these clinic characteristics showed a connection with the higher average’s number of NORs by nucleus and the comparison of NORs average’s number by nucleus on periphery bone fibroma and periphery giant cell lesion did not show a significant difference between the groups.

AgNOR polymorphism association with squamous intraepithelial lesions and invasive carcinoma with HPV infection / Asociación de los polimorfismos AgNORs con lesiones intraepiteliales escamosas, carcinoma cervical e infección por VPH

OBJECTIVE: Evaluate the relationships between AgNORs polymorphisms and squamous intraepithelial lesions (SIL) and squamous cell carcinoma (SCC) with HPV infection. MATERIALS AND METHODS: A study was carried out on sixty women from the state of Guerrero, Mexico. HPV detection was performed by PCR. AgNORs were identified by argentic impregnation. One hundred cells per slide were counted and classified according to the polymorphism of AgNORs dots; typical (spherical) and atypical (large, kidney-shaped and clustered). RESULTS: A total of 100 percent of the cases were positive for HPV infection. Nine different high-risk HPV genotypes were found, type16 was the most common (48.6 percent). The AgNORs showed a significant decrease in spherical shape according to neoplastic development. The three atypical shapes showed a significant increase in SIL and SCC (p-trend<0.001). CONCLUSIONS: AgNORs polymorphism rises progressively according to the grade of histological lesions that can be useful as a prognosis for progression of SCC.

OBJETIVO: Evaluar la relación entre los polimorfismos de AgNORs con las lesiones intraepiteliales escamosas (LIE) y carcinoma de células escamosas (CCE). MATERIAL Y MÉTODOS: Se estudiaron sesenta mujeres del estado de Guerrero, México. La detección del VPH fue por PCR y los AgNORs por impregnación argéntica; se contaron 100 células y se clasificaron por tipo de polimorfismo de AgNORs: típico (esférico) y atípicos (largo, forma de riñón o de racimo). RESULTADOS: El 100 por ciento de los casos presentaron infección por VPH, se encontraron nueve genotipos diferentes de VPH de alto riesgo, el 16 fue el más común (48.6 por ciento). La forma esférica de los polimorfismos de AgNORs mostró una disminución con el desarrollo neoplásico y las atípicas incrementaron progresivamente con SIL y SCC (p-tendencia<0.001). CONCLUSIONES: Los polimorfismos AgNORs se incrementan progresivamente con el grado de lesión histológica, y pueden ser útiles en el pronóstico de progresión del carcinoma cervical.

Astyanax hastatus Myers, 1928 (Teleostei, Characidae): a new species complex within the genus Astyanax?

Four populations of Astyanax hastatus Myers 1928 from the Guapimirim River basin (Rio de Janeiro State) were analyzed and three distinct cytotypes identified. These cytotypes presented 2n = 50 chromosomes, with 4M+8SM+10ST+28A (Cytotype A), 8M+10SM+14ST+18A (Cytotype B), 6M+8SM+4ST+32A (Cytotype C) and scanty heterochromatin, mainly located throughout pericentromeric regions of several chromosomal pairs. No homologies with the As-51 satellite DNA were observed in the three cytotypes, although all of them presented multiple 18S rDNA sites, as detected by both silver nitrate staining and FISH (fluorescent in situ hybridization). The application of the term "species complex" in Astyanax is discussed from a cytotaxonomic viewpoint.

Application of the comet assay in erythrocytes of Oreochromis niloticus (Pisces): a methodological comparison

The present study applied the comet assay to erythrocytes of Oreochromis niloticus with the aim of improving protocols to detect DNA damage in these cells, by using two distinct pHs (pH = 12.1 and pH > 13) and evaluating whether there is a correspondence between silver and ethidium bromide staining. Comets were visually examined and, the frequency of cells with and without damage was obtained, as well as the distribution of classes and scores. By using the Kruskal-Wallis test, our results revealed that pH 12.1 is more effective, although both pHs can be used. Our findings also suggest that silver staining can substitute ethidium bromide, an expensive and highly toxic stain that requires specific equipment for examination.

Pattern of silver nitrate-staining during meiosis and spermiogenesis in testicular lobes of Antiteuchus tripterus (Heteroptera: Pentatomidae)

The pattern of silver nitrate (Ag)-staining differed among testicular lobes of Antiteuchus tripterus. In general, these differences are in regard to the number, size, shape, coloring intensity, and location of the stained bodies or masses, observed during meiosis and spermiogenesis. These characteristics were similar in lobes 1-3. Lobes 4-6, however, differed from each other and from lobes 1-3 as well. Because the Ag-staining method is specific for nucleolar organizing regions and nucleolar material, the observations in meiosis of lobes 1-3 suggested the presence of a single pair of nucleolar organizing region-bearing chromosomes in A. tripterus, as previously found in other Pentatomidae species. In general, the amount of Ag-stained material seen in meiosis of the testicular lobes 1-3 of A. tripterus is smaller than in the other lobes. The differences among Ag-staining in testicular lobes of Antiteuchus tripterus lobes observed during spermiogenesis included a striking variation in morphology of the Ag-stained material found in the head and tail of the spermatids. Given that the key role of the nucleolar material is to participate in protein synthesis, interlobular variations seem to be related to the different functions attributed to each lobe (reproduction to lobes 1-3 and basically nutrition to lobes 4-6). To our knowledge, this is the first time that the nucleolar material was studied in each testicular lobe during spermatogenesis. The present observations encourage further studies since, in addition to being of basic biological interest, several Pentatomidae species are agricultural pests and added knowledge of their biology, mainly in reproduction, may be important for the development of control strategies.

Avaliação da cinética do epitélio corneano com marcadores de proliferação celular / Evaluation of the corneal epithelium kinetics using cell proliferation markers

OBJETIVO: Avaliar a cinética celular do epitélio corneano de coelhas em três situações: controle, hiperproliferação e hipoproliferação celular, com a utilização dos marcadores de proliferação celular BrdU, Ki-67/MIB-1 e AgNOR. MÉTODOS: Foram utilizadas quinze coelhas albinas que tiveram seus olhos aleatoriamente divididos em 3 grupos (A, B e C). O grupo A incluiu olhos que foram submetidos à instilação de tampão fosfato (total de 10 olhos); o grupo B, instilação de tampão fosfato após a remoção de uma área central do epitélio corneano de 10 mm (total de 10 olhos) e o grupo C, instilação de 5-fluoruracil em superfície ocular íntegra (total de 10 olhos). RESULTADOS: Os resultados da média e desvio-padrão do número de células marcadas pela BrdU nos grupos A, B e C foram, respectivamente, de 7,17 ± 0,74; 35,00 ± 3,01 e 0,22 ± 0,1 células marcadas por 100 células basais. As diferenças entre os grupos foram estatisticamente significantes. A média e o desvio padrão do número de células marcadas utilizando o Ki-67 foram de 7,55 ± 1,22 no grupo A; 35,55 ± 3,84 no grupo B e 0,34 ± 0,14 no grupo C. As diferenças entre os grupos foram estatisticamente significantes. A média e o desvio-padrão da medida da área das NORs no grupo A foram de 1,92 ± 0,24, no grupo B foram de 3,61 ± 0,27 e no grupo C foram de 1,71 ± 0,26. CONCLUSÕES: Os marcadores BrdU, Ki-67 e AgNOR apresentaram uma correlação positiva e estatisticamente significante nas situações de proliferação celular avaliadas (controle, hiperproliferação e hipoproliferação); o emprego do AgNOR não permitiu identificar diferenças na proliferação celular nas situações controle e hipoproliferação e houve maior concordância de resultados entre a BrdU e o Ki-67 nas três situações de proliferação celular.

PURPOSE: In order to maintain its clear and uniform structure, the corneal epithelium needs constant equilibrium between production (division) and desquamation of its epithelial cells. The author aimed to evaluate the cell kinetics of corneal epithelium of rabbits in three situations (control, hypoproliferation and hyperproliferation) using BrdU, Ki-67/MIB-1 and AgNORs proliferation markers. METHODS: Fifteen white female rabbits had their eyes randomly divided into three groups (A, B and. C). Group A included eyes submitted to phosphate buffer saline instillation (total 10 eyes); group B, instillation of phosphate buffer saline after removing a 10 mm central area of the corneal epithelium (total 10 eyes) and group C, instillation of 5-fluorouracil in normal epithelium (total 10 eyes). RESULTS: The results of the mean number and standard deviation of the marked cells using BrdU in groups A, B e C were, respectively, 7.17 ± 0.74; 35.00 ± 3.01 e 0.22 ± 0.1 marked cells per 100 basal cells. Differences among groups were statistically significant. The mean number and standard deviation of the labelled cells using Ki-67 were 7.55 ± 1.22 in group A; 35.55 ± 3.84 in group B and 0.34 ± 0.14 in group C. Differences among groups were statistically significant. The mean area and standard deviation of NORs in group A were 1.92 ± 0.24, in group B, 3.61 ± 0.27 and in group C, 1.71 ± 0.26. CONCLUSIONS: The markers BrdU, Ki-67 and AgNOR showed a positive correlation with statistical significance among the cellular proliferation situations studied (control, hypoproliferation and hyperproliferation); the AgNOR did not show statistically significant differences among the control and hypoproliferation situations and there was more agreement in the results among markers BrdU and Ki-67 in three cell proliferation situations.

Preponderance of GC-rich sites in silver-stained nucleolus organizing regions of Rita rita (Hamilton) and Mystus gulio (Hamilton) (Bagridae, Pisces), as revealed by chromomycin A3-staining technique and scanning electron microscopic studies

The karyotypes of two species of catfish, Rita rita (Hamilton) (2n = 54; 14m + 34sm + 6st; NF = 102) and Mystus gulio (Hamilton) (2n = 58; 30m + 12sm + 2st + 14t, NF = 100) were studied through Giemsa-, silver- and chromomycin A(3)-staining techniques. The silver-stained karyotypes in both sexes of R. rita and M. gulio revealed that the nucleolus organizing regions were located terminally at the shorter arms (Tp) of one pair of submetacentric chromosomes, placed at positions Nos. 2 and 1, respectively, which was confirmed by scanning electron microscopy. Staining with a GC-specific fluorochrome, chromomycin A(3), produced bright fluorescence in the Ag-positive nucleolus organizer regions, suggesting thereby that nucleolus organizing regions actually included GC-rich sites of active r-RNA genes in metaphase chromosomes of these two bagrids. Further such studies are needed due to the extreme paucity of data on fish.

Influence of two self-etching primer systems on enamel adhesion

The aim of this study was to compare two self-etching and a total-etch adhesive systems by assessing their shear bond strength to bovine enamel and the microleakage on class V composite restorations prepared on bovine enamel. Bovine teeth selected and allocated in three groups: Group 1: Scothbond Multi-Purpose; Group 2: Clearfil Liner Bond 2V; Group 3: Etch & Prime 3.0. For the microleakage test, each group was composed of ten class V restorations on the buccal surface. Two examiners attributed scores ranging from 0 (without leakage) to 3 (maximum leakage) to determine silver nitrate penetration at enamel-composite interface. Microleakage data were analyzed statistically by Kruskal-Wallis and Mann-Whitney tests at 5 percent significance level. For the bond strength test, ten teeth of each group were included, had their buccal surfaces flattened in order to obtain a 3-mm-diameter area to which a resin cylinder was bonded. After one week, the specimens were tested in shear strength at a crosshead speed of 0.5 mm/min. Bond strength data were treated by ANOVA and LSD tests at 5 percent significance level. The debonded interfaces were examined under scanning electron microscopy. No leakage was observed along enamel margins. Means (± SD) in MPa were: 18.75 (±5.83), 22.17 (±4.95) and 14.93 (±6.7) for Groups 1, 2, and 3, respectively. According to the results of this study, the self-etching primer systems presented statistically similar behavior (p>0.05) to that of the total-etch adhesive system (used as a control), not only regarding marginal leakage at bovine enamel-composite resin interface, but also regarding the shear bond strength of the bovine enamel. However, the self-etching primer systems differed significantly (p>0.05) to each other, with better results for Clearfil Liner Bond 2V. In conclusion, the self-etching primer systems had a performance comparable to that of the total-etch adhesive system.

O objetivo deste estudo foi comparar um sistema adesivo que utiliza condicionamento ácido total com outros contendo primers autocondicionantes em sua composição, quando aplicados ao esmalte bovino. Para isto, foram realizados testes de microinfiltração com corante e testes de resistência de união ao cisalhamento. Os dentes bovinos foram divididos em três grupos, de acordo com o sistema adesivo empregado: Grupo 1 (controle) - Scotchbond Multi-Uso Plus (3M); Grupo 2- Clearfil Liner Bond 2V (Kuraray); e Grupo 3- Etch & Prime 3.0 (Degussa). Para microinfiltração, cada grupo foi composto de dez restaurações classe V na superfície vestibular dos dentes e restauradas com o compósito TPH (Dentisply). A avaliação foi realizada por dois avaliadores previamente calibrados, usando um sistema de escore crescente de 0 (sem infiltração) até 3 (infiltração além da parede axial), dependendo do grau de penetração de um corante a base de nitrato de prata. Para o teste de resistência ao cisalhamento, 10 dentes de cada grupo foram incluídos em um tubo de metal, usando uma resina autopolimerizável. Suas superfícies vestibulares foram desgastadas até a obtenção de uma área de adesão com diâmetro de 3 mm. Sobre estas superfícies foram construídos cilindros de resina composta TPH. Após uma semana, foram submetidos ao ensaio de cisalhamento, a uma velocidade de 0,5 mm/min. Após a ruptura, as superfícies dos cilindros de resina composta foram recolhidas para observação em microscópio eletrônico de varredura. Para os testes de microinfiltração, os resultados foram tratados estatisticamente por Kruskal-Wallis e Mann-Whitney (p>0,05). O teste de microinfiltração não revelou diferença entre os grupos. Os resultados do teste de cisalhamento foram tratados por ANOVA e teste LSD (p<0,05). As médias e desvios padrão em MPa foram: G1: 18,75 ± 5,83; G2: 22,17 ± 4,95; G3: 14,93 ± 6,7. Não houve diferença estatisticamente significante entre o grupo controle e os autocondicionantes...

Evaluation of nucleolar organizer regions in maxillary osteosarcoma

Maxillary osteosarcomas are a relatively frequent malignant tumor of the oral cavity. Similarly to other skeletal osteosarcomas, they exhibit different cellular differentiation patterns, i.e. chondroblastic, osteoblastic, or fibroblastic. Although their histological features resemble those of osteosarcomas of the long bones, their pattern of evolution usually differs. Morphometric variations in silver stained Nucleolar Organizer Regions (AgNOR) have proved of value to study the biology of several tumors. However, information on the analysis of AgNOR in maxillary tumors is scarce. The aim of the present study was to analyze the variations of different morphological parameters related to AgNOR in a series of 32 cases of maxillary osteosarcoma. In each case we analyzed 100 nuclei corresponding to the prevalent cellular differentiation type, selecting the most aggressive area. We employed software previously developed at our laboratory that yields information on different AgNOR-related parameters. The results were compared with those previously reported in a study on 12 cases of osteosarcoma of long bones. Six cases of oral mucosa squamous cell carcinoma were also included for comparative purposes. Single AgNOR volume proved to be the most discriminatory and informative parameter. The value of single AgNOR volume was considerably lower in mandible osteosarcomas than in osteosarcomas of the upper maxilla (p=0.02). The values were significantly lower in maxillary osteosarcomas than in long bone osteosarcomas and in oral carcinomas. This finding would suggest a slower rate of cell activity in maxillary osteosarcomas, associated in turn to its known lower degree of aggressiveness. The present results suggest that the analysis of AgNOR is a valuable and easily applicable marker to determine the degree of malignancy and biology of maxillary osteosarcomas.

Los osteoscaromas de maxilares son entidades relativamente frecuentes entre los tumores malignos de la cavidad bucal. Al igual que los osteosarcomas de otras localizaciones del esqueleto, pueden presentar diferentes patrones de diferenciación celular (condroblástico, osteoblástico o fibroblástico). Si bien sus características histológicas son similares, tienen generalmente un comportamiento evolutivo diferente al de los huesos largos. Las variaciones morfométricas de las regiones organizadoras del nucleoloidentificadas por impregnación argéntica (AgNOR) han demostrado ser marcadores útiles para el estudio de la biología de diversas entidades tumorales, pero hay muy escasa información de su análisis en tumores de los huesos maxilares. El objetivo de este trabajo fue analizar las variaciones de diferentes parámetros morfológicos de las AgNOR en una serie de 32 casos de osteosarcomas de maxilar. En cada caso se analizaron 100 núcleos en el patrón de diferenciación celular predominante, seleccionando la zona de mayor agresividad. Se utilizó un programa que aporta información sobre diferentes parámetros de AgNOR, desarrollado previamente en nuestro laboratorio. El parámetro más indicativo resultó ser el volumen individual de las AgNOR. Este parámetro en los osteosarcomas con localización mandibular fue considerablemente menor que aquellos localizados en maxilar superior (p=0.02). En los osteosarcomas de maxilar los valores fueron significativamente menores que en los de huesos largos y en los carcinomas bucales. Ellos podría ser indicativo de una menor actividad celular, a su vez asociada a su reconocida menor agresividad. Estos resultados sugieren que el análisis de AgNOR podría ser considerado como un marcador de utilidad y de fácil aplicación para determinar el grado de malignidad en osteosarcomas de maxilar y estimar su comportamiento biológico.