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Despite of the existence of previous knowledge on the relative growth of U. maracoani for a population of southeastern Brazilian coast, the aim of the study was described and compare the relative growth patterns and sexual maturity of this species from Formoso River mangrove, state of Pernambuco, northeast Brazil, with data from other locations and of congeneric species. The crabs were monthly sampled manually, with a capture effort by one person colleting for 30 minutes at the low tide from April 2009 to March 2010. Crabs of both sexes were measured with a caliper at following dimensions: carapace width (CW), carapace length (CL), cheliped propodus length (CPL), cheliped propodus height (CPH) and abdomen width (AW). Additionally, males were also measured at their gonopod length (GL). The determination of morphological maturity was based on the relationship between the CPL vs. CW for males and AW vs. CW for females, which were the relations that best evidenced the beginning of the morphological sexual maturity for U. maracoani. The relationships CPL vs. CW and AW vs. CW evidenced positive allometry for juveniles and adult’s males and females. However, a negative allometry was observed in the relationship CL vs. CW and GL vs. CW for juveniles and adult’s males, AW vs. CW for adults’ males and, CPH vs. CW for adults’ females. The growth pattern shown by the female abdomen probably is related to egg incubation and the excessive growth observed at the male cheliped is probably related to reproductive process, because this appendage is used at the courtship and intra or inter specific agonistic interactions with other males. The morphological sexual maturity of males was estimated at 15.8 mm CW (50% of males), with the smallest mature male measuring 17.34 mm, and the largest immature male 18.15 mm CW. Females reached morphological sexual maturity (50%) at 13.8 mm CW, with the smallest mature female at 15.43 mm CW and the largest immature female at 16.32 mm CW. The differential growth patterns between juvenile and adult phases, as well as the onset sexual maturity seem to have relationship with ontogenetic changes, which is mainly related to behavioral or reproductive traits. The growth pattern observed in the female's abdomen is probably related to hatching eggs while overgrowth shown in male cheliped should be related to reproductive processes, as this appendix is used in court and in specific interactions with other males.

Moringa oleifera Lam. leaves infusion and powder are widely used by population due the nutritional and medicinal potentials, however data regarding safety of use are still inconclusive, leading to prohibition of this plant in some countries.The present work investigated the nutritional and phytochemical composition, acute and 28-day repeated dose toxicity, and genotoxicity of M. oleifera leaves infusion and powder.For nutritional characterization of leaf powder, it was determined: humidity; mineral residue (ash); total lipid, protein, carbohydrate, and crude fiber contents; and total caloric value. Phytochemical composition was determined by high performance liquid chromatography (HPLC). The acute toxicity assay used Swiss female albino mice and oral administration in a single dose at 2000 and 5000 mg/kg of infusion or powder. The 28-day repeated dose toxicity assay employed female and male mice, with oral administration of infusion or powder at the doses 250, 500 and 1000 mg/kg. The animals were evaluated for body weight, water and feed consumption, biochemical and hematological parameters, and histology of the liver, spleen, and kidneys. In vivo genotoxicity and mutagenicity (2000 mg/kg) were evaluated by the comet assay and the micronucleus test, respectively.Nutritional characterization confirmed that M. oleifera leaves are rich in proteins, carbohydrates, lipids, minerals, and fiber. HPLC indicated the presence of flavonoids and cinnamic derivatives as major polyphenols. Acute toxicity did not reveal alterations in weight gain and water and feed consumptions and no change in biochemical, hematological, and histological parameters. Behavior alterations was observed in the first 2 h after administration at 5000 mg/kg in both treatments. Infusion did not present toxicity when administered for 28 days. Conversely, the powder at 500 and 1000 mg/kg promoted liver and kidney damages observed through biochemical parameters and histopathology. Genotoxicity and mutagenicity were not detected at 2000 mg/kg.The present study reveals that M. oleifera leaves are an important source of polyphenols and nutrients. Indiscriminate use of both infusion and crude leaf powder above 2000 mg/kg and powder at 500 and 1000 mg/kg are not recommended. Chronic toxicological studies and establishment of preparation protocols are suggested aiming to guarantee the safety in the use of M. oleifera leaves as nutraceutical by population.

O OBJETIVO DESTE TRABALHO FOI APLICAR UMA SEQUÊNCIA DIDÁTICA (SD) SOBRE A ORIGEM DA VIDA, NO ENSINO REMOTO, UTILIZANDO O TIKTOK PARA PRODUÇÃO DE VÍDEO EDUCATIVO PELOS ALUNOS. A INTERVENÇÃO FOI REALIZADA EM UMA ESCOLA DA REDE ESTADUAL DE ENSINO DE GRAVATÁ-PE, COM ALUNOS DO ENSINO MÉDIO. BASEADA NA ABORDAGEM DO ENSINO POR INVESTIGAÇÃO, A SD FOI DIVIDIDA EM ORIENTAÇÃO, INVESTIGAÇÃO, COMUNICAÇÃO DOS RESULTADOS E DIVULGAÇÃO DOS RESULTADOS. TODAS AS ETAPAS DO PROCESSO FORAM AVALIADAS CONSTANTEMENTE PELA PROFESSORA; UM QUESTIONÁRIO SOBRE O CONTEÚDO FOI APLICADO ANTES E APÓS A SD E OS RESULTADOS FORAM COMPARADOS ENTRE SI. FOI POSSÍVEL VERIFICAR QUE HOUVE APRENDIZADO E PARTICIPAÇÃO ATIVA DOS ALUNOS NO DESENVOLVIMENTO DAS ATIVIDADES. ASSIM, ASSOCIAR UMA SD DE CARÁTER INVESTIGATIVO A UMA TECNOLOGIA DE COMUNICAÇÃO CONHECIDA PELOS ADOLESCENTES PODE REPRESENTAR UMA FORMA DE CONTRIBUIR COM SUA APRENDIZAGEM, TORNANDO A AULA REMOTA MAIS INTERATIVA, E PROMOVER AUTONOMIA E PROTAGONISMO DOS ALUNOS.

Scorpion envenomation represents an important health problem in many parts of the world, due to the high number and severity of accidents. Recent studies demonstrated that some species can produce venoms with genetic damage potential. Here, we evaluated whether Tityus stigmurus venom causes genetic damage in blood and testicular cells of Swiss mice. We also analyzed the effect of the venom on the number of spermatogenic lineage cells. Five groups of mice received 0.387 mg/kg of the venom, intraperitoneally; one group received saline solution (control group). Blood and testicular cells were collected for comet assay and histological analysis at different times after treatment (1, 2, 6, 12, and 48 h). Blood was also collected 48 h after treatment for the micronucleus test in erythrocytes. Histological analysis was performed by counting cells of the spermatogenic lineages; the nuclear area of elongated spermatocytes was also evaluated. Treatment with the venom induced DNA damage that endured from 1 h to 48 h, as confirmed by the comet assay. The micronucleus test demonstrated that the venom induced mutations in erythrocytes. The number of spermatogonia and rounded spermatids decreased in some groups; the number of elongated spermatids increased, and their nuclear size decreased 1 h after treatment. Genetic damage can be caused directly by the venom, but we suggested that reactive oxygen species that result from inflammatory process caused by the envenomation may have an important role in the DNA damage. Genetic damage and apoptosis may explain the changes in the number of spermatogenic cells. Furthermore, the decrease in nuclear area may result from chromatin loss. Genetic damage in testicular cells, associated with alterations in the number and morphology of spermatogenic cells, can result in reproduction disorders in animals, or humans, stung by T. stigmurus.

Ethnopharmacological relevance Morus alba L. (white mulberry) is used in traditional medicine worldwide, including Brazil. The leaves of this plant are used to treat inflammatory disorders. Universal interest in this plant necessitates studies on the toxicological safety and scientific substantiation of the medicinal properties of M. alba. In previous work, we investigated the acute toxicity of orally administered M. alba ethanol extract in mice. Aim of the study This work was designed to investigate the ethanol extract obtained from M. alba leaves for acute toxicity when intraperitoneally administered, in vivo genotoxicity, and potential to reduce acute inflammation. In order to further investigate the constituents of the extract, we also obtained the high-performance liquid chromatography (HPLC) fingerprint of the extract. Materials and methods Phytochemical analysis by thin layer chromatography (TLC) was performed and the results were used to obtain the HPLC fingerprint. Acute toxicity of 300 and 2000 mg/kg b.w. i.p. doses administered to mice for 14 days was evaluated. Genotoxicity was evaluated by counting the number of micronucleated polychromatic erythrocytes in the blood of mice that either received or did not receive the extract at 75, 150 and 300 mg/kg b.w. per os. The anti-inflammatory effect of the same doses administered per os was investigated using the carrageenan air pouch model. Results The TLC analysis of the extract revealed the presence of a remarkable amount of flavonoids and cinnamic acids. The HPLC fingerprint showed the presence of one major peak corresponding to chlorogenic acid and two smaller peaks corresponding to flavonoids. In the toxicity assays, there were no deaths or deviations in behavior of treated mice as compared to the control at any dose. However, biochemical, hematological, and histological analyses showed that intraperitoneal injection caused several forms of damage to the mice, which were not observed in case of oral administration, studied in our previous work. Oral administration of the extract did not result in genotoxicity and considerably reduced (58.6–65.6% inhibition) leukocyte migration in all doses evaluated, in comparison with the negative control. Conclusions The ethanol extract from M. alba leaves administered intraperitoneally possesses a greater degree of toxicity in mice when compared to per os administration. The extract was not genotoxic when ingested by mice and exhibited a highly inhibitory effect against acute inflammation, which is probably linked to the presence of chlorogenic acid and flavonoids in the composition. This work contributes to the determination of safety of the medicinal use of M. alba leaves.

The objective of this study was to determine the cytotoxicity of organic extracts of P. moniliformis in vitro and identify the acute toxicity and genotoxicity in vivo. The leaves were extracted using three organic solvents (cyclohexane [EP1], ethyl acetate [EP2], and methanol [EP3]). Phytochemical qualitative analysis was performed by thin layer chromatography (TLC). Cytotoxicity tests were performed on human embryonic kidney (HEK) cells and J774 murine macrophages. Acute toxicity in mice was measured after intraperitoneal (ip) administration of 2000 mg/kg, while evaluation of genotoxicity and mutagenicity were assessed using the comet assay and the micronucleus (MN) test, respectively. The TLC analysis of the extracts revealed the presence of flavonoids, triterpenes, steroids, and saponins. In the cytotoxicity assay, extracts EP1 and EP3 altered proliferation of HEK cells, and all organic extracts increased the viability of J774 cells. In the toxicity tests, no deaths or behavioral alterations were observed in mice exposed to the acute dose of the extracts. Although some extracts led to changes in hematological and histological parameters, these results did not indicate physiological changes. In relation to the MN test and comet assay, no significant changes were detected in the DNA of the animals tested with the extracts EP1, EP2, and EP3. Thus, extracts of P. moniliformis were not considered to be toxic and did not induce formation of MN or damage to cellular DNA in the genotoxicity tests.

Exposure of pregnant women to pesticides in an occupational or incidental manner has been pointed out as a possible cause of health problems in newborns. Cypermethrin and methomyl are insecticides whose active compounds and metabolites can cause harm to the health of pregnant women and their offspring. Both contribute to increases in the concentrations of oxidizing agents that damage DNA, causing mutations. Thus, the use of antioxidant substances, such as melatonin, could serve as protection against genetic damage, reducing the problems caused by insecticides. Here, we evaluated the protective effect of exogenous melatonin in adult rats and their offspring exposed to methomyl and cypermethrin during the gestational period. Thirty adult rats were divided into five groups from the first day of pregnancy: NC - negative control, Mth received 0.02 mg/kg/day of methomyl, Cyp received 0.05 mg/kg/day cypermethrin, MthMl received 0.02 mg/kg/day methomyl combined with 10 mg/kg/day melatonin, and CypMl received 0.05 mg/kg/day cypermethrin combined with 10 mg/kg/day melatonin. Blood and liver samples were collected from the dams; blood, liver, and brain samples were collected from the offspring. All tissues were used for comet assay; blood was used for micronucleus test. Our results demonstrate that cypermethrin and methomyl induce significant genetic damage in the tissues of dams and offspring. Treatment with melatonin significantly reduced cypermethrin-induced genetic damage in dams and pups; however, melatonin was not effective against the genetic damage caused by methomyl. Thus, melatonin may be considered a promising agent for the reduction of genotoxic effects induced by cypermethrin but appears to be inefficient against the genotoxic effects caused by methomyl, probably due to differences in the metabolism of each pesticide.

Background: Secondary metabolites are responsible for most of the biological activities of lichens. Many of these compounds exhibit significant antineoplastic activity. The aim of the present in vitro study was to evaluate the cytotoxic and genotoxic activities of organic extracts and purified barbatic acid from the lichen Cladonia salzmannii (Nyl.). Methods: The thallus of the lichen (22 g) was cleaned and dried with the solvents diethyl ether, chloroform and acetone. Organic extracts were obtained using the hot exhausted method in a Soxhlet apparatus. Barbatic acid (BAR) was purified from the ether extract (1.3 g). Chemical analysis of the organic extracts and purified BAR was performed using thin-layer chromatography. The purity of purified BAR was determined using high-performance liquid chromatography. The MTT method [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and cytokinesis-block proliferation index (CBPI) were used to determine the cytotoxic activity of the organic extracts and purified BAR. The micronucleus test and comet assay were used to determine genotoxic potential of the organic extracts and purified BAR. Dimethyl sulfoxide was used as the diluting solvent of the samples in all biological tests. Results: The IC50 results demonstrated significant cytotoxic potential of the ether extract (50 µg/mL) against cell lines NCI-H292 (IC50: 29.91 µg/mL), HEp-2 (IC50: 26.75 µg/mL) and HL-60 (IC50: 3.59 µg/mL) as well as the purified BAR (25 µg/mL) against cell lines HEp-2 (IC50: 15.79 µg/mL) and MCF-7 (IC50: 18.28 µg/mL). The CBPI demonstrated the cytotoxic activity of the purified BAR at all concentrations tested (5, 10, 20 and 40 µg/mL) and all organic extracts (50 µg/mL) against Ehrlich carcinoma cells. For sarcoma 180, only BAR purified at a concentration of 40 µg/mL and the ether and chloroform extracts (50 µg/mL) were considered cytotoxic. The micronucleus test revealed that the purified BAR at a concentration of 5 µg/mL had no genotoxic potential against either tumor cell line. Furthermore, the chloroform extract and purified BAR at a concentration of 10 µg/mL were not considered genotoxic for sarcoma 180. In the comet assay, all compounds tested induced DNA damage in both tumor lines. Conclusion: Based on the present results, organic extracts and purified barbatic acid from C. salzmannii exhibit cytotoxic and genotoxic activity against of the tumor cell lines tested.

Ethnopharmacological relevance Himatanthus drasticus is a tree popularly known as janaguba. Endemic to Brazil, it is found in the Cerrado and Caatinga biomes, rock fields, and rainforests. Janaguba latex has been used in folk medicine for its antineoplastic, anti-inflammatory, analgesic, and antiallergic activities. However, studies investigating the safety of its use for medicinal purposes are limited. Aim of the study: This study aimed to evaluate the toxicity of the latex extracted from H. drasticus. Materials and methods The latex was extracted from H. drasticus specimens by removing a small area of bark (5 × 30 cm) and then dissolving the exudate in water and lyophilizing it. Phytochemical screening was performed by TLC and GC-MS, protein, and carbohydrate levels. Cell viability was performed by the MTT method. Acute oral toxicity, genotoxicity, and mutagenicity assays were performed in mice. Results TLC showed the presence of saponins and reducing sugars, as well as steroids and terpenes. The GC-MS analysis of the nonpolar fraction identified lupeol acetate, betulin, and α/β-amyrin derivatives as the major compounds. The latex was toxic to S-180 cells at 50 and 100 μg/mL. No signals of toxicity or mutagenicity was found in mice treated with 2000 mg/kg of the latex, but genotoxicity was observed in the Comet assay. Conclusions H. drasticus latex showed toxicity signals at high doses (2000 mg/kg). Although the latex was not mutagenic to mice, it was genotoxic in the Comet assay in our experimental conditions. Even testing a limit dose of 2000 mg/kg, which is between 10 to 35-fold the amount used in folk medicine, caution must be taken since there is no safe level for genotoxic compounds exposure. Further studies on the toxicological aspects of H. drasticus latex are necessary to elucidate its possible mechanisms of genotoxicity.

Pregnant women may be exposed to pesticides in an occupational or incidental way, which has been pointed out as a possible cause of problems in the health of newborns. Cypermethrin and methomyl are insecticides whose active compounds and metabolites can cause harm to the health of pregnant women and the concept. Both increase the concentration of oxidizing agents that damage DNA, causing mutations. Thus, the use of antioxidant substances such as melatonin could serve as protection against genetic damage, reducing the problems caused by insecticides. Here, we evaluated the protective effect of exogenous melatonin in adult rats and their offspring exposed to methomyl and cypermethrin during the gestational period. Thirty adult rats were divided into five groups from the first day of pregnancy: NC - negative control; Mth received 0.02 mg / kg / day of methomyl; Cyp received 0.05 mg / kg / day cypermethrin; MthMl received 0.02 mg / kg / day methomyl combined with 10 mg / kg / day melatonin; CypMl received 0.05 mg / kg / day cypermethrin combined with 10 mg / kg / day melatonin. Blood and liver samples were collected from the dams; blood, liver, and brain samples were also collected from the offspring. All tissues were analysed in the comet assay; while blood was also used for the micronucleus test. The results demonstrate that cypermethrin and methomyl induce significant genetic damage in the tissues of dams and offspring. Treatment with melatonin reduced Cyp-induced genetic damage in dams and pups; but was not effective against DNA damage caused by methomyl. Thus, melatonin may be considered a promising agent for the reduction of genotoxic effects induced by cypermethrin, but appears to be inefficient against the genotoxic effects caused by methomyl, probably due to differences in the metabolism of each pesticide.

Maternal alcoholism can induce serious injuries in embryonic and fetal development. The metabolism of alcohol increases the production of free radicals and acetaldehyde, molecules capable of reacting with DNA, impairing organogenesis. Melatonin is a powerful antioxidant that can act as a protective agent against DNA damage caused by genotoxic agents, such as ethanol. This study evaluated the protective effect of exogenous melatonin in rats and their offspring on the genotoxic response induced by chronic alcohol consumption during pregnancy. Twenty-five pregnant rats were divided into the following groups: NC - Negative control; ET - Rats receiving ethanol (3 g/kg/day); ET+10 M - Rats receiving ethanol (3 g/kg/day) and melatonin (10 mg/kg/day); ET+15 M - Rats receiving ethanol (3 g/kg/day) and melatonin (15 mg/kg/day); PC - Positive control (40 mg/kg cyclophosphamide). The dams and 10 pups (five males and five females) from each group were anesthetized to collect blood and liver from the dams and blood, liver and brain of neonates to evaluate the frequency of DNA damage by the comet assay. Blood was also used for the micronucleus test. The results demonstrated a significant increase in DNA damage in the blood and liver cells of dams receiving ethanol and their offspring as well as in the brain of these neonates. Treatments with melatonin (10 and 15 mg/kg/day) significantly reduced the genotoxicity caused by ethanol in the blood of dams and neonates (males and females), liver of dams and male offsprings, and in the brain of female offsprings. It was shown that only the female offspring exposed to maternal alcohol consumption showed a higher frequency of micronuclei in polychromatic erythrocytes. Consequently, exogenous melatonin may be a promising therapeutic agent against genotoxic damage induced by alcohol; however, further studies are needed to confirm these benefits.

Soft corals show a wide variety of reproductive strategies, including both asexual and sexual reproduction which can influence macro-evolutionary processes. The octocoral Carijoa riisei has an ample geographical distribution in Pacific and southern Atlantic and also in Caribbean region. This species was considered invasive in Hawaii, and its capacity for rapid proliferation is recognized, but recent studies, however, have indicated that it appears to be native to the Indo-Pacific region. The present study examined the reproductive biology of C. riisei in the Atlantic basin. The results were then compared to previous studies of the same species from the Pacific (Hawaii) and Caribbean (Puerto Rico) regions to examine the hypothesis that the reproductive patterns of C. riisei populations are the same throughout its geographical distribution, independent of its native or non-native status. Samples were collected on a monthly basis from May/2007 to April/2008 at Porto de Galinhas (Pernambuco State, Brazil). This species was found to have similar reproduction patterns in Brazil, the Caribbean, and in Hawaii (a gonochoric reproductive pattern and continuous and asynchronous gamete release). The similarities of their traits contribute to its rapid proliferation and occupation of spaces left by other species, independent of its native or non-native status.

The development of sex in sea turtles is determined by the temperature incubation of the eggs (TSD - Temperature dependent Sex Determination). For Eretmochelys imbricata species, very little is known about the sex ratio of spawning which occur in the southern coast of Pernambuco. Therefore, the purpose of this study is to analyze the relationship between sexual development and temperature incubation of nests of hawksbill turtles deposited in Merepe Beach County Ipojuca, Pernambuco. The temperature data were collected from nine nests during the months from January to June 2013, with the aid of registers type temperature data loggers (Onset Computer Model, UA‑001‑08). The average recorded for nests sampled depending on the incubation period and temperature were 54 days and 31.68°C. Based on these results, the nests showed a strong trend towards the development of females, with the sex ratio, estimated by histological analysis, 86.53% for females. So the beach Merepe has temperatures that provide greater differentiation of newborn females. However, more detailed studies should be carried out with the intention of verifying whether these results can be obtained for any ipojucano coast. O desenvolvimento do sexo em tartarugas marinhas é determinado pela temperatura de incubação dos ovos (TSD - Temperature-dependent Sex Determination). Para a espécie Eretmochelys imbricata, pouco se sabe a respeito da proporção sexual das desovas que ocorrem no litoral sul de Pernambuco. Diante disso, o objetivo desse estudo é analisar a relação entre o desenvolvimento sexual e a temperatura de incubação de ninhos de tartarugas-de-pente, depositados na Praia de Merepe, município do Ipojuca, Pernambuco. Os dados de temperatura foram coletados em nove ninhos, durante os meses de janeiro a junho de 2013, com auxílio de registradores de temperatura do tipo Data loggers (Modelo Onset Computer, UA-001-08). As médias registradas para os ninhos amostrados em relação ao período de incubação e temperatura foram de 54 dias e de 31,68ºC. Com base nesses resultados, os ninhos apresentaram uma forte tendência para o desenvolvimento de fêmeas, sendo a razão sexual, estimada por meio de análises histológicas, de 86,53% para fêmeas. Portanto, a Praia de Merepe possui temperaturas que propiciam uma maior diferenciação de neonatos fêmea. Entretanto, estudos mais detalhados devem ser realizados com a intenção de verificar se esses mesmos resultados podem ser obtidos para todo o litoral ipojucano.

Tonsillar lymphoid polyps are uncommon lesions that have rarely been studied. The authors describe the clinical, histopathologic, and immunohistochemical features of 6 tonsillar polyps in which lymphoid tissue represented more than 80% of the lesion. Presenting symptoms were tonsillar mass and/or dysphagia. No predisposing factor was detected. Microscopically, all polyps contained follicles with germinal centers, crypts lined by lymphoepithelium, and a small amount of fibrous tissue in the center of the lesion. B cells (CD20+), T cells (CD45RO+), plasma cells (κ+ and λ+) and vessels (lymphatic, D2-40+; blood, CD34+) presented distribution and architectural patterns as expected for lymphoid tissue of a palatine tonsil. Tonsillar lymphoid polyps are possibly hamartomas characterized by overgrowth of lymphoid elements, which maintain an architectural pattern and cellular composition similar to those of the palatine tonsil.

Summary Objective Human herpesvirus 8 (HHV-8) and Epstein–Barr virus (EBV) are human pathogens associated to a number of neoplasms, including tumors of the Waldeyer's ring. Both viruses have been previously detected by in situ methods in tonsils and adenoids from children. HHV-8 was found in 6.8% of the cases and EBV in about one third of the cases. As they belong to the same γ-herpesvirus subfamily and share some biological characteristics, it is of medical interest to further explore their possible relationship in the Waldeyer's ring, an issue not yet addressed in the specialized literature. The purpose of the present study is to compare the presence of EBV by in situ hybridization (ISH) in tonsils and adenoids from children up to 14 years of age in cases previously shown to be positive and negative for HHV-8. Methods Paraffin wax-embedded sections consisting of 38 tonsils and two adenoids from 40 patients were analyzed. HHV-8 was detected by ISH, using the T1-1 probe for the viral mRNA. EBV was also detected by ISH, using the EBER probe. Both probes and the detection systems were provided by Novocastra. Results HHV-8 was detected in 19 tonsils and one adenoid. The other 19 tonsils and one adenoid taken from the HHV-8-negative group were selected by pairing age and gender of patients with the HHV-8-positive group. In both groups EBV was detected in 13 cases and was negative in other 7. Conclusion Although both viruses are related in many aspects, some biological and epidemiological features differ. This is reflected in the present results, as EBV is similarly detected in the groups negative and positive for HHV-8, favoring different mechanisms of spread.

Mycosis fungoides (MF) is the most common form of primary cutaneous T-cell lymphoma. In its early stage it may mimic benign dermatoses both on a clinical and histologic basis. MF usually expresses CD3 and CD4 (T-cell) markers. CD7 is expressed on about 90% of CD4 T cells and is often deficient on malignant T cells. Thus, CD7 may be useful in evaluating the nature of dermal lymphoid infiltrates. The aim of this study was to evaluate the usefulness of immunohistochemical detection of T-cell markers on paraffin-embedded sections, CD3 and CD7 (clone CBC.37), in the differential diagnosis of MF and benign dermatoses. Forty-two patients with diffuse dermal T-lymphocytic infiltrates were selected. Previous clinicopathologic correlation and follow-up had established the diagnosis of MF in 31 patients and benign dermatoses in 11. The mean value of stained cells in MF was 86.45% for CD3 and 53.09% for CD7 (P

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:43:19Z No. of bitstreams: 1 S1676-24442005000400008.pdf: 1103487 bytes, checksum: fc025297b6caa4496aed850598e4c310 (MD5) Made available in DSpace on 2013-08-22T18:43:19Z (GMT). No. of bitstreams: 1 S1676-24442005000400008.pdf: 1103487 bytes, checksum: fc025297b6caa4496aed850598e4c310 (MD5) Previous issue date: 2005-08-01 Made available in DSpace on 2013-09-30T19:30:57Z (GMT). No. of bitstreams: 2 S1676-24442005000400008.pdf: 1103487 bytes, checksum: fc025297b6caa4496aed850598e4c310 (MD5) S1676-24442005000400008.pdf.txt: 27143 bytes, checksum: 324c50b70653661a5d03b60af984199d (MD5) Previous issue date: 2005-08-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:15:53Z No. of bitstreams: 2 S1676-24442005000400008.pdf: 1103487 bytes, checksum: fc025297b6caa4496aed850598e4c310 (MD5) S1676-24442005000400008.pdf.txt: 27143 bytes, checksum: 324c50b70653661a5d03b60af984199d (MD5) Made available in DSpace on 2014-05-20T13:15:53Z (GMT). No. of bitstreams: 2 S1676-24442005000400008.pdf: 1103487 bytes, checksum: fc025297b6caa4496aed850598e4c310 (MD5) S1676-24442005000400008.pdf.txt: 27143 bytes, checksum: 324c50b70653661a5d03b60af984199d (MD5) Previous issue date: 2005-08-01 A técnica de imuno-histoquímica é usada na rotina diagnóstica e na pesquisa em patologia humana desde 1970, porém seu uso na patologia veterinária é relativamente recente, principalmente com objetivo diagnóstico. A maior dificuldade no uso da imuno-histoquímica na patologia veterinária tem sido a falta de anticorpos específicos para os tecidos animais. Na falta de anticorpos específicos para as espécies domésticas, a patologia veterinária freqüentemente faz uso de anticorpos que apresentam reatividade cruzada entre antígenos humanos e animais. O objetivo deste trabalho foi testar a reatividade cruzada de diversos anticorpos feitos para uso humano em tecido parafinado de algumas espécies animais, utilizando-se dos novos métodos de recuperação antigênica e amplificação da reação imuno-histoquímica. No presente estudo foi possível confirmar a aplicabilidade de que muitos anticorpos produzidos para diagnóstico imuno-histoquímico em patologia humana podem ser utilizados em patologia veterinária. Novos estudos são necessários a fim de se ampliar a lista de aplicabilidade desses anticorpos em diferentes espécies animais, levando sempre em consideração as variações de clones, diluições, métodos de recuperação antigênica e de revelação. Immunohistochemistry has become a practical and widely used tool for diagnosis in human pathology since the 70's. However, its application in veterinary diagnostic pathology has not been so common, especially due to the lack of specific antibodies. To overcome this drawback, antibodies which present cross reactivity with human and animal antigens have been applied. The purpose of the present study was to test the cross reactivity of some antibodies intended for the human pathology, which may be used in animal tissues, with the help of antigen retrieval and amplification systems. In the present study it was confirmed that many of the antibodies produced for use in human histopathology might be applied in veterinary pathology. Further studies are needed to increase the list of applicability of these antibodies to different animal species. It must be stressed that in this type of study some variables, such as clone of antibody, dilution, antigen retrieval method, and detection system, have to be evaluated. Universidade Estadual de Campinas Faculdade de Ciências Médicas Department of Anatomic Pathology Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinárias Department of Veterinary Pathology UNICAMP Centro de Atenção Integral à Saúde da Mulher Laboratory of Experimental Pathology Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinárias Department of Veterinary Pathology

Summary Objective Human herpesvirus 8 (HHV8) has been associated with multicentric Castleman's disease, Kaposi's sarcoma and effusion non-Hodgkin's lymphoma. Epidemiological studies have shown seropositivity in variable proportions of populations. It seems to be sexually transmitted among adults and through oral contact among children. The virus has been demonstrated in desquamating oral epithelial cells, but there is no report on its presence in the Waldeyer's ring. The purpose of the present study is to detect HHV8 in tonsils and adenoids from children up to 20 years of age in which these organs had been surgically removed due to hypertrophy, using immunohistochemistry and in situ hybridization. Methods Paraffin wax-embedded sections consisting of 181 tonsils and 162 adenoids from 293 patients were analyzed. HHV8 was detected by immunohistochemistry (IHC) using the anti-LNA1 antibody (Novocastra) and the LSAB+ detection system (Dako). For the in situ hybridization (ISH), the T1-1 probe for the viral mRNA and the detection system used were provided by Novocastra. Results In 20 cases (6.83%), HHV8 was detected in cells morphologically characterized as lymphoid. In three of them epithelial cells were also positive. In 19 cases, the virus was detected in tonsils and in just 1 case in an adenoid. In all 20 cases detection was possible by ISH, whereas in only 2 of them there was a concomitant positivity by IHC. Conclusion Our data support the oral route of contamination by HHV8 in children, in whom tonsils and adenoids may harbor the virus. It is found especially in tonsils and only rarely in adenoids. In these organs, ISH is the method of choice to detect this virus, probably due to the small amount of viral proteins.

1. A human glioma cell line, NG97, was established by Grippo et al. in 2001 from tissue obtained from a grade III astrocytoma (WHO, 2000). In this first study, the cell line grew as two morphologically distinct subpopulations: dendritic/spindle cells and small round cells. The injection of NG97 cells into nude mice induced an aggressive tumor characterized by: severe cytological atypia, vascular proliferation and pseudopalisading necrosis (glioblastoma multiforme features). 2. The purpose of the present study was to characterize the immunophenotype and ultrastructural aspects of this cell line, using the parental tumor, cultured cells and the xenotransplant, in order to assess its glial nature and possible divergent differentiation. 3. NG97 cells and xenotransplant expressed the main neuroglial markers (GFAP, S-100 protein, NSE and Leu-7) and showed no aberrant expression of other histogenetic markers. GFAP was similarly expressed in the parental tumor and in the cells in culture, but decreased in the xenotransplant. NSE expression was reduced in NG97 cells, but substantially recovered in the xenotransplant. This variability in expression of GFAP and NSE was interpreted as either a phenomenon of dedifferentiation or to microenvironmental selection of specific subclones. S-100 was equally expressed in the three contexts. The xenotransplant’s ultrastructural features were those of a highly undifferentiated tumor. No significant immunophenotypic or ultrastructural differences between the two morphologically distinct populations were found. 4. Thus, our data demonstrate that NG97 cells constitute a pure glial-committed cell line, which may prove useful as a malignant glioma model in studies addressing pathophysiological, diagnostic and therapeutic issues.

The importance of in situ immunodetection of hormone receptors for therapy planning and prognostic evaluation in patients with breast carcinoma is well established. Sensitive detection methods are of utmost importance, especially in poorly fixed tissues, which are not uncommon in routine pathologic practice. The purpose of the present study is to compare immunoexpression of estrogen receptors in 20 cases of invasive ductal carcinoma using two antibodies, 1D5 and 6F11, and to verify the effect of different antigen retrieval solutions and detection systems. Immunoperoxidase was performed on paraffin sections using 1D5 and 6F11 as primary antibodies. Heat-induced antigen retrieval was performed using citrate buffer (pH 6.0) or Tris-EDTA buffer (pH 8.9). Detection was achieved using the following systems: EnVision, EnVision Plus, and labeled streptavidin-biotin peroxidase complex. Reaction was semiquantified from 0 to 4. There were no differences between the two markers, 1D5 and 6F11, except when 6F11 was used with EnVision and citrate buffer, in which case weaker reactivity was observed. Only in this combination (6F11/EnVision) was EDTA buffer significantly better than citrate. Labeled streptavidin-biotin peroxidase complex presented the best results, followed by EnVision Plus.

Summary Objective: Herpes simplex virus (HSV) has been described as cause of acute tonsillitis. It has also been found in nasopharyngeal florid lymphoid infiltrate, mostly composed of CD4+, CD56+ T-cells, simulating lymphoma. In spite of its widespread prevalence in latent form, to the best of our knowledge no study is available on in situ detection of HSV in chronically hyperplastic nasopharyngeal lymphoid tissue. The purpose of the present study was to search for the presence of HSV 1 and 2 in 21 adenoids and 15 tonsils from children (2–12 years of age) in which these organs had been surgically removed due to hypertrophy. Methods: Paraffin wax-embedded sections from the 36 cases were submitted to the in situ hybridization technique, using the biotinilated probe to Herpes simplex virus 1 and 2 (Pan Path, Amsterdam) and the Rembrandt ® Universal DISH & HRP Detection Kit (Pan Path, Amsterdam). Positive control consisted of a previously tested Herpes infected lung. Results: In none of the 36 cases studied were positive nuclei detected in adenoid and tonsils, either in lymphoid, in stroma or in epithelial cells, as those seen in the positive control. Conclusion: HSV does not seem to be implied in tonsil or adenoid chronic lymphoid hyperplasia. These organs do not seem to harbor the virus latently, or the amount of virus is too low to be detected without amplification methods.