Influência das endometrites nas estruturas ovarianas e qualidade oocitária em tratos reprodutivos post-mortem de vacas / Influence of endometritis on ovarian structures and oocyte quality in post mortem reproductive tract of cows

Objetivou-se no presente estudo estimar o percentual de endometrites clínicas (EC) e subclínicas (ES) em tratos reprodutivos post-mortem de fêmeas bovinas, verificando sua influência sobre as estruturas ovarianas, alterações presentes no ovário, número e qualidade de oócitos recuperados. Foram coletados e avaliados 171 tratos reprodutivos em matadouro frigorífico. As endometrites foram diagnosticadas por meio de citologia endometrial e pela presença e características das secreções uterinas. Os diagnósticos foram confirmados pela análise histopatológica, sendo avaliada a existência de infiltrados de células inflamatórias no endométrio. Os ovários foram avaliados macroscopicamente quanto à estrutura e alterações ovarianas. A qualidade oocitária foi avaliada de acordo com o número de camadas de células do cumulus e aspecto do citoplasma, sendo assim, classificados como: grau I (GI), grau II (GII), grau III (GIII) e grau IV (GIV). Dentre as peças avaliadas, 8,20% (n = 14) apresentaram EC e 4,10% (n = 7) foram diagnosticadas com ES. Observou-se que a presença de endometrites não exerceu efeito sobre as estruturas e alterações ovarianas, assim como não alterou o número de oócitos recuperados. Em relação ao grau de qualidade oocitária, foi encontrado variação entre os diferentes graus de qualidade dentro dos grupos, no entanto, não houve correlação aparente com a presença de infecção uterina.(AU)

The objective of this study was to estimate the percentage of clinical (CE) and subclinical (SE) endometritis in post mortem bovine reproductive tracts, analyzing their influence on ovarian structures, changes in the ovary, and number and quality of oocytes collected. Endometritis were diagnosed by endometrial cytology, and by the presence and characteristics of uterine secretions. The diagnosis was confirmed through histopathological analysis, by evaluating the presence of infiltrated inflammatory cells in the endometrium. The ovaries were macroscopically evaluated for presence of physiological structures and alterations. The oocyte quality evaluation was performed according to the number of cumulus cells layers and cytoplasmic aspect, being classified as: grade I (GI), grade II (GII), grade III (GIII) and grade IV (GIV). Among the uterus evaluated, 8,20% (n = 14) of the animals presented EC, and 4,10% (n = 7) were diagnosed with ES. It was observed that the presence of endometritis had no effect on ovarian structures and alterations, nor did it influence the number of oocytes collected. Regarding the degree of oocyte quality, a variation was found between the different grades of quality within the groups, however, there was no apparent correlation with the presence of uterine infection.(AU)

Influência das endometrites nas estruturas ovarianas e qualidade oocitária em tratos reprodutivos post-mortem de vacas / Influence of endometritis on ovarian structures and oocyte quality in post mortem reproductive tract of cows

Objetivou-se no presente estudo estimar o percentual de endometrites clínicas (EC) e subclínicas (ES) em tratos reprodutivos post-mortem de fêmeas bovinas, verificando sua influência sobre as estruturas ovarianas, alterações presentes no ovário, número e qualidade de oócitos recuperados. Foram coletados e avaliados 171 tratos reprodutivos em matadouro frigorífico. As endometrites foram diagnosticadas por meio de citologia endometrial e pela presença e características das secreções uterinas. Os diagnósticos foram confirmados pela análise histopatológica, sendo avaliada a existência de infiltrados de células inflamatórias no endométrio. Os ovários foram avaliados macroscopicamente quanto à estrutura e alterações ovarianas. A qualidade oocitária foi avaliada de acordo com o número de camadas de células do cumulus e aspecto do citoplasma, sendo assim, classificados como: grau I (GI), grau II (GII), grau III (GIII) e grau IV (GIV). Dentre as peças avaliadas, 8,20% (n = 14) apresentaram EC e 4,10% (n = 7) foram diagnosticadas com ES. Observou-se que a presença de endometrites não exerceu efeito sobre as estruturas e alterações ovarianas, assim como não alterou o número de oócitos recuperados. Em relação ao grau de qualidade oocitária, foi encontrado variação entre os diferentes graus de qualidade dentro dos grupos, no entanto, não houve correlação aparente com a presença de infecção uterina.

The objective of this study was to estimate the percentage of clinical (CE) and subclinical (SE) endometritis in post mortem bovine reproductive tracts, analyzing their influence on ovarian structures, changes in the ovary, and number and quality of oocytes collected. Endometritis were diagnosed by endometrial cytology, and by the presence and characteristics of uterine secretions. The diagnosis was confirmed through histopathological analysis, by evaluating the presence of infiltrated inflammatory cells in the endometrium. The ovaries were macroscopically evaluated for presence of physiological structures and alterations. The oocyte quality evaluation was performed according to the number of cumulus cells layers and cytoplasmic aspect, being classified as: grade I (GI), grade II (GII), grade III (GIII) and grade IV (GIV). Among the uterus evaluated, 8,20% (n = 14) of the animals presented EC, and 4,10% (n = 7) were diagnosed with ES. It was observed that the presence of endometritis had no effect on ovarian structures and alterations, nor did it influence the number of oocytes collected. Regarding the degree of oocyte quality, a variation was found between the different grades of quality within the groups, however, there was no apparent correlation with the presence of uterine infection.

Are grape juices more erosive than orange juices?

AIMS: To evaluate the chemical characteristics of grape and orange juices, and their erosive potential in the decrease of microhardness and the loss of enamel structure. METHODS: Five grape and orange juices were evaluated for pH, titratable acidity, calcium, phosphate, and fluoride concentration. De-ionised water and Cola soft drink were used as a negative and positive control, respectively. Twelve specimens of bovine enamel were immersed in beverages for 10 min at 37 °C, 3 times/day for 7 days. Erosive potential was quantified using microhardness and loss of enamel structure. Anova One Way, Student's t test, Multiple Regression and Spearman Correlation (p < 0.05) were used to analyse the results. RESULTS: Powdered grape juice showed the lowest pH (3.18 ± 0.03) and pure grape juice presented the highest titratable acidity (5.48 ± 0.06 mL NaOH/100 mL). Fresh orange juice and soya-based grape juice revealed the lowest calcium (0.77 ± 0.12 mmol/L) and phosphate concentrations (0.35 ± 0.06 mmol/L), respectively. Among juices, powdered orange juice caused the greatest decrease in surface microhardness (SMH) (127.99 ± 40.47 ΔSMH) and grape juice from concentrate caused the greatest loss of enamel structure (13.30 ± 3.56 µm). CONCLUSIONS: All of the evaluated juices contributed to dental erosion. Grape juices presented greater erosive potential than orange juices. Pure, powdered and concentrated grape juices showed similar loss of enamel structure to the Cola soft drink. The erosive potential of beverages was statistically correlated to pH, titratable acidity, calcium, phosphate and fluoride concentrations.

Dynamic flux of microvesicles modulate parasite-host cell interaction of Trypanosoma cruzi in eukaryotic cells.

Extracellular vesicles released from pathogens may alter host cell functions. We previously demonstrated the involvement of host cell-derived microvesicles (MVs) during early interaction between Trypanosoma cruzi metacyclic trypomastigote (META) stage and THP-1 cells. Here, we aim to understand the contribution of different parasite stages and their extracellular vesicles in the interaction with host cells. First, we observed that infective host cell-derived trypomastigote (tissue culture-derived trypomastigote [TCT]), META, and noninfective epimastigote (EPI) stages were able to induce different levels of MV release from THP-1 cells; however, only META and TCT could increase host cell invasion. Fluorescence resonance energy transfer microscopy revealed that THP-1-derived MVs can fuse with parasite-derived MVs. Furthermore, MVs derived from the TCT-THP-1 interaction showed a higher fusogenic capacity than those from META- or EPI-THP-1 interaction. However, a higher presence of proteins from META (25%) than TCT (12%) or EPI (5%) was observed in MVs from parasite-THP-1 interaction, as determined by proteomics. Finally, sera from patients with chronic Chagas disease at the indeterminate or cardiac phase differentially recognized antigens in THP-1-derived MVs resulting only from interaction with infective stages. The understanding of intracellular trafficking and the effect of MVs modulating the immune system may provide important clues about Chagas disease pathophysiology.

COMPARISON OF AGGLUTINATING AND GROWTH INHIBITION ANTIBODIES LEVELS IN SOWS IMMUNIZED WITH TWO EXPERIMENTAL INACTIVATED MONOVALENT VACCINES OF CANICOLA SEROVAR: WHOLE CULTURE WITH ALUMINUM HYDROXIDE AND LIPOPOLYSACCHARIDE SUBUNIT WITH LIPID A

ABSTRACT It was compared the antibody response of sows immunized with two experimental vaccines produced with L.interrogans, serovar Canicola, strain LO-4, isolated in Brazil.One of the vaccines was the usual bacterin (whole culture inactivated with phenol and adjuvanted with aluminum hydroxide -WC-AlOH3) and the other one was a subunit vaccine produced with a lipopolysaccharide (LPS) fraction extracted from the bacteria outer envelop and with the lipid A, also extracted from the leptospira wall as adjuvant (LPS-MPLA). Experiment was as follows: group 1 (n = 11), not immunized control, group 2, (n = 11): two immunization with 30 days interval of LPS-MPLA vaccine and group 3 (n = 11): two immunization with 30 days interval of WC-AlOH3 vaccine All three groups were simultaneously immunized, independently of pregnancy stage. Both agglutinin and neutralizing post vaccination antibodies levels were measured respectively by the microscopic sera agglutination with live antigens test (MAT) and the in vitro leptospira growth inhibition test (GIT). Sera collections were performed each 30 days during four months after the first vaccination. Non vaccinated control group animals presented no agglutinating antibodies against Canicola serovar during the whole experiment. At 32 and 68 post vaccination days the agglutinating antibodies levels of group 2 (LPS-MPLA) were significantly higher than the observed in group 3 (WC AlOH3), respectively, p = 0.013 and p = 0.031. The differences observed in the growth inhibition antibodies titers of the two vaccines tested were not significant (p > 0.05). Despite the peak of post-vaccination agglutinins have been registered at 68 days after first immunization, higher levels growth inhibition antibodies were detected at 30 days of first vaccination. Subunit vaccine presented the same immunogenic capacity for the production of neutralizing antibodies as the whole culture one.

RESUMO Foi comparada a resposta imune de fêmeas suínas adultas imunizadas contra a leptospirose, com vacinas monovalentes produzidas com L.interrogans, sorovar Canicola estirpe LO4, isolada no Brasil. A vacina foi empregada em duas formas: cultura de bactérias totais inativada e acrescida do adjuvante de hidróxido de alumínio (WC-AlOH3) e a do tipo de subunidade constituída apenas por uma fração de lipopolisacarídios (LPS) extraídos do envelope externo da bactéria tendo como adjuvante o monofosforil lipídio A, também extraído da parede da leptospira (LPS-MPLA). O delineamento experimental incluiu: grupo 1 (n = 11): controle não imunizado; grupo 2 (n = 11): imunizado com duas aplicações em intervalo de 30 dias da vacina LPS MFLA; Grupo 3 (n = 11): imunizado com duas aplicações em intervalo de 30 dias da vacina WC-AlOH3. Todos os grupos foram imunizados simultaneamente sem ser considerado o estágio de gestação dos animais. Os níveis de anticorpos pós-vacinais, aglutinantes e neutralizantes foram avaliados, respectivamente, pelos testes de soroaglutinação microscópica com antígenos vivos (SAM) e o de inibição do crescimento de leptospiras in vitro (ICLIV). O monitoramento sorológico foi efetuado a cada 30 dias durante quatro meses após aplicação da primeira dose da vacina. Os animais do grupo controle, não vacinados, não apresentaram anticorpos aglutinantes para o sorovar Canicola durante todo o período experimental. Aos 32 e 68 dias da primo-vacinação, os níveis de anticorpos aglutinantes do grupo 2 (LPS-MPLA) foram significativamente superiores aos observados no grupo 3 (WC AlOH3), respectivamente p = 0,013 e p = 0,031. As diferenças observadas nos níveis de anticorpos inibidores do crescimento de leptospiras in vitro, induzidos pelas duas vacinas, não foram significativas (p > 0,05). A despeito do pico de anticorpos aglutinantes pós-vacinais ter sido registrado aos 68 dias da primeira imunização, os níveis mais elevados de anticorpos inibidores do crescimento de leptospiras já foram observados aos 30 dias da primo-vacinação. A vacina de subunidade apresentou a mesma capacidade de indução de anticorpos neutralizantes que a vacina de bactérias totais.

Comparison of agglutinating and growth inhibition antibodies levels in sows immunized with two experimental inactivated monovalent vaccines of Canicola serovar: whole culture with aluminum hydroxide and lipopolysaccharide subunit with lipid A / Comparação dos níveis de anticorpos pós-vacinais, aglutinantes e inibidores do crescimento de leptospiras em fêmeas suínas imunizadas com duas vacinas inativadas experimentais monovalentes do sorovar Canicola: cultivo total inativado com hidróxido de alumínio e subunidade de lipopolissacarídios com monofosforil lipídio A

It was compared the antibody response of sows immunized with two experimental vaccines produced with L.interrogans, serovar Canicola, strain LO-4, isolated in Brazil.One of the vaccines was the usual bacterin (whole culture inactivated with phenol and adjuvanted with aluminum hydroxide -WC-AlOH3) and the other one was a subunit vaccine produced with a lipopolysaccharide (LPS) fraction extracted from the bacteria outer envelop and with the lipid A, also extracted from the leptospira wall as adjuvant (LPS-MPLA). Experiment was as follows: group 1 (n = 11), not immunized control, group 2, (n = 11): two immunization with 30 days interval of LPS-MPLA vaccine and group 3 (n = 11): two immunization with 30 days interval of WC-AlOH3 vaccine All three groups were simultaneously immunized, independently of pregnancy stage. Both agglutinin and neutralizing post vaccination antibodies levels were measured respectively by the microscopic sera agglutination with live antigens test (MAT) and the in vitro leptospira growth inhibition test (GIT). Sera collections were performed each 30 days during four months after the first vaccination. Non vaccinated control group animals presented no agglutinating antibodies against Canicola serovar during the whole experiment. At 32 and 68 post vaccination days the agglutinating antibodies levels of group 2 (LPS-MPLA) were significantly higher than the observed in group 3 (WC AlOH3), respectively, p = 0.013 and p = 0.031. The differences observed in the growth inhibition antibodies titers of the two vaccines tested were not significant (p > 0.05). Despite the peak of post-vaccination agglutinins have been registered at 68 days after first immunization, higher levels growth inhibition antibodies were detected at 30 days of first vaccination. Subunit vaccine presented the same immunogenic capacity for the production of neutralizing antibodies as the whole culture one.

Foi comparada a resposta imune de fêmeas suínas adultas imunizadas contra a leptospirose, com vacinas monovalentes produzidas com L.interrogans, sorovar Canicola estirpe LO4, isolada no Brasil. A vacina foi empregada em duas formas: cultura de bactérias totais inativada e acrescida do adjuvante de hidróxido de alumínio (WC-AlOH3) e a do tipo de subunidade constituída apenas por uma fração de lipopolisacarídios (LPS) extraídos do envelope externo da bactéria tendo como adjuvante o monofosforil lipídio A, também extraído da parede da leptospira (LPS-MPLA). O delineamento experimental incluiu: grupo 1 (n = 11): controle não imunizado; grupo 2 (n = 11): imunizado com duas aplicações em intervalo de 30 dias da vacina LPS MFLA; Grupo 3 (n = 11): imunizado com duas aplicações em intervalo de 30 dias da vacina WC-AlOH3. Todos os grupos foram imunizados simultaneamente sem ser considerado o estágio de gestação dos animais. Os níveis de anticorpos pós-vacinais, aglutinantes e neutralizantes foram avaliados, respectivamente, pelos testes de soroaglutinação microscópica com antígenos vivos (SAM) e o de inibição do crescimento de leptospiras in vitro (ICLIV). O monitoramento sorológico foi efetuado a cada 30 dias durante quatro meses após aplicação da primeira dose da vacina. Os animais do grupo controle, não vacinados, não apresentaram anticorpos aglutinantes para o sorovar Canicola durante todo o período experimental. Aos 32 e 68 dias da primo-vacinação, os níveis de anticorpos aglutinantes do grupo 2 (LPS-MPLA) foram significativamente superiores aos observados no grupo 3 (WC AlOH3), respectivamente p = 0,013 e p = 0,031. As diferenças observadas nos níveis de anticorpos inibidores do crescimento de leptospiras in vitro, induzidos pelas duas vacinas, não foram significativas (p > 0,05). A despeito do pico de anticorpos aglutinantes pós-vacinais ter sido registrado aos 68 dias da primeira imunização, os níveis mais elevados de anticorpos inibidores do crescimento de leptospiras já foram observados aos 30 dias da primo-vacinação. A vacina de subunidade apresentou a mesma capacidade de indução de anticorpos neutralizantes que a vacina de bactérias totais.

Follow-up of the development of femoral degeneration lesions in broilers / Seguimiento del desarrollo de lesiones por degeneración en pollos de engorde

A study was carried out in the experimental facilities of FMVZ/UNESP-Botucatu, with the aim of following-up the development and the incidence of femoral degeneration (FD). A total of 305 one-day-old male broilers were housed in six pens of 5m² each. Histological analyses of femur head collected when broilers were 0, 7, 14, 21, 28, 35, and 42 days of age were carried out. At 42 days of age, 30 birds were taken to the experimental processing plant of FMVZ for leg gross examination. Ten legs per FD score where selected, and histologically analyzed to determine the most probable age at the beginning of the lesions, and to standardize femoral degeneration lesion scores. The histological results showed that cell architecture started to disorganize at 21 days of age in the resting and proliferation zones, and that angiogenesis increased, invading the joint cartilage, The gross lesion indexes due to femoral degeneration were 22.5 percent, 42.5%, and 65% at 28, 35, and 42 days of age, respectively.

Se realizó un estudio en las instalaciones experimentales de FMVZ/UNESP-Botucatu, con el objetivo de seguir el desarrollo y la incidencia de degeneración femoral (DF) en pollos. Se utilizaron 305 polluelos de un día, machos, distribuidos en seis corrales de 5m² cada uno. Se analizaron cortes histológicos de cabezas de fémur recolectadas a los 0, 7, 14, 21, 28, 35 y 42 días de edad. A los 40 días de edad, se llevaron 30 aves al Matadero Experimental de FMVZ, para análisis macroscópico de las piernas. Se escogieron 10 muslos por escore de DF, y se analizaron histológicamente para determinar la edad más probable del inicio de la lesión y estandarizar los escores de lesión por degeneración femoral. Los resultados histológicos indicaron que a los 21 días ocurre el inicio de la desorganización celular en la zona de reposo y de proliferación, además del aumento de la angiogénesis, invadiendo el cartílago articular. Microscópicamente, el índice de lesión por degeneración femoral fue del 22.5 por ciento, 42.5% y 65% a los 28, 35 y 42 días de edad, respectivamente.

Femur mineral density of broilers with femoral degeneration fed high nutritional density diets / Densidad mineral del fémur de pollos de engorde con degeneración femoral alimentados con dietas de alta densidad nutricional

A study was carried out in the experimental facilities of FMVZ/UNESP-Botucatu, with the aim of following-up the development and the incidence of femoral degeneration (FD). A total of 305 one-day-old male broilers were housed in six pens of 5m² each. A completely randomized experimental design, with 3 treatments (T1traditional nutritional density diet; T2high nutritional density diet) of 3 replicates each was applied. Femoral head of the broilers were submitted to gross examination at 0, 7, 14, 21, 28, 35, and 42 days of aged. At 42 days of age, 60 birds (30 per treatment) were submitted to the Veterinary Hospital of FMVZ to determine bone mineral density by radiography. Birds were then sacrificed for gross examination of the legs, and FD scoring. Five legs per treatment within each FD score were submitted to computed tomography for femur head integrity and bone mineral density. Treatments did not influence FD incidence, and the first gross FD lesions appeared when birds were 28 days old. It was concluded that radiographic optical densitometry and computed tomography are efficient methods to evaluate femoral degeneration, and both techniques expressed the same profile. In addition, using radiographic optical densitometry and computed tomography, these results also allowed us to establish bone mineral density value ranges within each gross FD score. These finding may provide an excellent non-invasive tool to describe femoral degeneration.

Se realizó un estudio en las instalaciones experimentales de FMVZ/UNESP-Botucatu, con el objetivo de seguir el desarrollo y la incidencia de degeneración femoral en pollos. Se utilizaron 305 polluelos de un día, machos, distribuidos en seis corrales de 5m² cada uno. Se adoptó un delineamiento experimental totalmente al azar, con dos tratamientos de 3 repeticiones cada uno. Se alimentaron las aves del T1 con dietas con densidad nutricional convencional, mientras el T2 consistió de una dieta con alta densidad nutricional. Se realizaron análisis macroscópicos de la cabeza del fémur de aves de 0, 7, 14, 21, 28, 35 y 42 días de edad. A los 42 días de edad, se llevaron 60 aves (30 por tratamiento) al Hospital Veterinario de FMVZ, para hacer radiografías para el análisis de la densidad mineral ósea. Posteriormente, se sacrificaron los pollos para el análisis macroscópico de las piernas y se atribuyeron puntajes para DF. Se seleccionaron cinco muslos por tratamiento dentro de cada puntaje de DF, que fueron sometidas a tomografía para evaluación de la integridad y de la densidad ósea de la cabeza del fémur. Los tratamientos no tuvieron influencia en la incidencia de DF, y a partir de los 28 días de vida, las aves presentaron lesiones macroscópicas. Se estableció que la densitometría ósea y la tomografía son métodos eficaces para evaluar la DF, además que ambos expresan el mismo perfil. Por otra parte, se encontraron intervalos de valores para densidad mineral ósea obtenida por densitometría óptica radiográfica y por tomografía en función de los puntajes macroscópicos de DF. Esos hallazgos son una importante herramienta no invasiva para la caracterización de degeneración femoral.

Erosive effect of an antihistamine-containing syrup on primary enamel and its reduction by fluoride dentifrice.

AIM: This study evaluated the action of an antihistamine-containing syrup (Claritin D) on enamel that was subsequently submitted or not to applications of fluoride dentifrice. METHODS; Two hundred sixty-four slices (n=44 per subgroup) prepared from exfoliated primary molars were evaluated in hardness tests. Six subgroups were submitted to different treatments for 10 days. The controls underwent pH cycling with (positive control) or without (negative control) three daily immersions in fluoride dentifrice/distilled water slurry. The test subgroups related to daytime use of the antihistamine syrup underwent pH cycling and two 5-min applications of Claritin D, coupled or not to the three daily immersions in the fluoride slurry. The subgroups related to nocturnal use of the syrup were submitted to the same procedures of daytime subgroups, respectively, but with one of the applications of Claritin D lasting for 8 h. RESULTS: The median hardness values obtained after use of the syrup were significantly lower than the initial ones. Equivalent values for subgroups submitted to fluoride applications in addition to treatment with the syrup were significantly higher. CONCLUSION: It was concluded that the antihistamine-containing syrup reduced the hardness of primary enamel and that, in this experiment, the use of fluoride dentifrice was able to diminish this erosive effect.

Human autoantibodies to diacyl-phosphatidylethanolamine recognize a specific set of discrete cytoplasmic domains.

The aim of this study was to characterize a novel human autoantibody-autoantigen system represented as cytoplasmic discrete speckles (CDS) in indirect immunofluorescence (IIF). A distinct CDS IIF pattern represented by 3-20 discrete speckles dispersed throughout the cytoplasm was identified among other cytoplasmic speckled IIF patterns. The cytoplasmic domains labelled by human anti-CDS-1 antibodies did not co-localize with endosome/lysosome markers EEA1 and LAMP-2, but showed partial co-localization with glycine-tryptophan bodies (GWB). CDS-1 sera did not react with several cellular extracts in immunoblotting and did not immunoprecipitate recombinant GW182 or EEA1 proteins. The typical CDS-1 IIF labelling pattern was abolished after delipidation of HEp-2 cells. Moreover, CDS-1 sera reacted strongly with a lipid component co-migrating with phosphatidylethanolamine (PE) in high performance thin-layer chromatography (HPTLC)-immunostaining of HEp-2 cell total lipid extracts. The CDS-1 major molecular targets were established by electrospray ionization-mass spectrometry (ESI-MS), HPTLC-immunostaining and chemiluminescent enzyme-linked immunosorbent assay as diacyl-PE species, containing preferentially a cis-C18 : 1 fatty acid chain at C-2 of the glycerol moiety, namely 1,2-cis-C18 : 1-PE and 1-C16 : 0-2-cis-C18 : 1-PE. The clinical association of CDS-1 sera included a variety of systemic and organ-specific autoimmune diseases but they were also observed in patients with no evidence of autoimmune disease.

[Prevalence, treatment needs, and predisposing factors for traumatic injuries to permanent dentition in 11-13-year-old schoolchildren]. / Prevalência, necessidade de tratamento e fatores predisponentes do traumatismo na dentição permanente de escolares de 11 a 13 anos de idade.

The objective of this study was to assess the prevalence of traumatic injuries to the permanent incisors and the association with clinical predisposing factors and parents' schooling. A cross-sectional survey was conducted with schoolchildren aged 11 to 13 years in Biguaçu, Brazil. Dental examinations were conducted by a dentist, and the criteria for traumatic dental injuries used in the children's dental health survey in the United Kingdom were adopted. the study recorded the type of damage sustained, treatment performed or needed, the size of incisal overjet, and whether lip coverage was adequate. Socio-demographic data included sex, age, and parents' level of schooling. a total of 2,260 children were examined, and prevalence rates were 10.4%, 10.6%, and 11.2% in 11, 12, and 13-year-old children, respectively. Treatment need was 6.3 interventions per thousand incisors. Male gender and overjet greater than 5mm were significantly related to having a traumatic dental injury. Inadequate lip coverage and parents' educational level were not associated with dental trauma. The study concluded that male gender and incisal overjet greater than 5mm are associated with the occurrence of dental injury.

Prevalência, necessidade de tratamento e fatores predisponentes do traumatismo na dentição permanente de escolares de 11 a 13 anos de idade / Prevalence, treatment needs, and predisposing factors for traumatic injuries to permanent dentition in 11-13-year-old schoolchildren

Este estudo objetivou determinar a prevalência do traumatismo dentário na dentição permanente e observar associações com fatores predisponentes. Foi realizado um estudo transversal com escolares de 11 a 13 anos de Biguaçu, Santa Catarina, Brasil. Os exames foram realizados por um cirurgião-dentista utilizando os mesmos critérios do Children's Dental Health Survey do Reino Unido. Foram observados tipo de dano, tratamento providenciado e necessidade, overjet incisal, adequabilidade da cobertura labial, idade, sexo e nível de educação dos pais. Foram examinadas 2.260 crianças e as prevalências encontradas foram: 10,4 por cento, 10,6 por cento e 11,2 por cento aos 11, 12 e 13 anos, respectivamente. A necessidade de tratamento foi de 6,3 incisivos por mil examinados. Escolares do sexo masculino e com overjetmaior que 5mm tiveram mais traumatismo dentário do que escolares do sexo feminino e com overjetincisal até 5mm. Cobertura labial inadequada e nível de educação dos pais não estiveram estatisticamente associados com o traumatismo dentário. Concluiu-se que ser do sexo masculino ou ter um overjetincisal maior do que 5mm aumenta a chance de sofrer traumatismo dentário.

Multiserotype protection of mice against pneumococcal colonization of the nasopharynx and middle ear by killed nonencapsulated cells given intranasally with a nontoxic adjuvant

Intranasal challenge of C57BL/6 mice with Streptococcus pneumoniae serotypes 6B, 14, and 23F produced colonization of the middle ear and NP. Intranasal vaccination with ethanol-killed nonencapsulated cells with adjuvant protected both sites. Of four nontoxic adjuvants tested, the cholera toxin B subunit was most effective and least nonspecifically protective

On the biosynthesis of Rhodnius prolixus heme-binding protein.

The biosynthesis of Rhodnius prolixus heme-binding protein (RHBP), which is present in the hemolymph and oocytes of Rhodnius prolixus, was investigated. Fat bodies of female insects incubated in vitro with 14C-leucine were able to synthesize and secrete 14C-RHBP to the culture medium. Titrtion of synthesized RHBP with hemin showed that the protein secreted by the fat bodies is bound to heme, despite the presence of apo-RHBP in the hemolymph. The sequence of the RHBP cDNA encodes a pre-protein of 128 amino acids with no significant homology to any known protein. Northern-blot assays revealed that RHBP expression was limited to fat bodies. The levels of both RHBP mRNA and secreted protein increased in response to blood meal. In addition, the time-course of RHBP secretion in vitro paralleled mRNA accumulation observed in vivo. The inhibition of the de novo heme biosynthesis by treatment of fat bodies with succinyl acetone (SA), an irreversible inhibitor of delta-aminolevulinic acid-dehydratase, led to a significant decrease of heme-RHBP secretion. Nevertheless, the levels of RHBP mRNA were not modified by SA treatment, suggesting that the heme availability is involved in a post-transcriptional control of the RHBP synthesis.

Proinflammatory activity of glycosylphosphatidylinositol anchors derived from Trypanosoma cruzi: structural and functional analyses.

A strong activation of macrophages is observed during acute infection with Trypanosoma cruzi. Little is known, however, about the parasite molecules that are responsible for this early activation of innate immunity. Recent studies have shown the stimulatory activity of protozoan-derived glycosylphosphatidylinositol (GPI) anchors on cultured macrophages. In this review, we provide a detailed analysis of the correlation between structure and proinflammatory activity by T. cruzi-derived GPI anchors. We also cover the studies that have identified the Toll-like receptor 2 as a functional GPI receptor and have partially characterized signaling pathways triggered by T. cruzi-derived GPI anchors, which lead to the synthesis of proinflammatory cytokines in macrophages. Finally, we discuss the implications of these findings in resistance and pathogenesis during the infection with T. cruzi.

Activation of Toll-like receptor-2 by glycosylphosphatidylinositol anchors from a protozoan parasite.

Glycosylphosphatidylinositol (GPI) anchors and glycoinositolphospholipids (GIPLs) from parasitic protozoa have been shown to exert a wide variety of effects on cells of the host innate immune system. However, the receptor(s) that are triggered by these protozoan glycolipids has not been identified. Here we present evidence that Trypanosoma cruzi-derived GPI anchors and GIPLs trigger CD25 expression on Chinese hamster ovary-K1 cells transfected with CD14 and Toll-like receptor-2 (TLR-2), but not wild-type (TLR-2-deficient) Chinese hamster ovary cells. The protozoan-derived GPI anchors and GIPLs containing alkylacylglycerol and saturated fatty acid chains or ceramide were found to be active in a concentration range of 100 nM to 1 microM. More importantly, the GPI anchors purified from T. cruzi trypomastigotes, which contain a longer glycan core and unsaturated fatty acids in the sn-2 position of the alkylacylglycerolipid component, triggered TLR-2 at subnanomolar concentrations. We performed experiments with macrophages from TLR-2 knockout and TLR-4 knockout mice, and found that TLR-2 expression appears to be essential for induction of IL-12, TNF-alpha, and NO by GPI anchors derived from T. cruzi trypomastigotes. Thus, highly purified GPI anchors from T. cruzi parasites are potent activators of TLR-2 from both mouse and human origin. The activation of TLR-2 may initiate host innate defense mechanisms and inflammatory response during protozoan infection, and may provide new strategies for immune intervention during protozoan infections.

Requirement of mitogen-activated protein kinases and I kappa B phosphorylation for induction of proinflammatory cytokines synthesis by macrophages indicates functional similarity of receptors triggered by glycosylphosphatidylinositol anchors from parasitic protozoa and bacterial lipopolysaccharide.

In the present study, we evaluated the ability of GPI-anchored mucin-like glycoproteins purified from Trypanosoma cruzi trypomastigotes (tGPI-mucin) to trigger phosphorylation of different mitogen-activated protein kinases (MAPKs) and related transcription factors in inflammatory macrophages. Kinetic experiments show that the peak of extracellular signal-related kinase (ERK)-1/ERK-2, stress-activated protein kinase (SAPK) kinase-1/mitogen-activated protein kinase (MAPK) kinase-4, and p38/SAPK-2, phosphorylation occurs between 15 and 30 min after macrophage stimulation with tGPI-mucin or GPI anchors highly purified from tGPI-mucins (tGPI). The use of the specific inhibitors of ERK-1/ERK-2 (PD 98059) and p38/SAPK-2 (SB 203580) phosphorylation also indicates the role of MAPKs, with possible involvement of cAMP response element binding protein, in triggering TNF-alpha and IL-12 synthesis by IFN-gamma-primed-macrophages exposed to tGPI or tGPI-mucin. In addition, tGPI-mucin and tGPI were able to induce phosphorylation of I kappa B, and the use of SN50 peptide, an inhibitor of NF-kappa B translocation, resulted in 70% of TNF-alpha synthesis by macrophages exposed to tGPI-mucin. Finally, the similarity of patterns of MAPK and I kappa B phosphorylation, the concentration of drugs required to inhibit cytokine synthesis, as well as cross-tolerization exhibited by macrophages exposed to tGPI, tGPI-mucin, or bacterial LPS, suggest that receptors with the same functional properties are triggered by these different microbial glycoconjugates.

The Rubino test for leprosy is a beta2-glycoprotein 1-dependent antiphospholipid reaction.

We describe the isolation and identification of three components required for the Rubino reaction (RR), which is the rapid sedimentation of formalinized sheep red-blood cells (SRBC) initiated by serum from leprosy patients with defective Mycobacterium leprae-specific cell immunity. The Rubino reaction factor (RRF) required for this phenomenon, previously identified as an immunoglobulin M (IgM), was purified from leprosy patient serum by adsorption to formalinized SRBC. Purified RRF IgM, when added to formalinized SRBC, did not produce a positive RR. However, when the contact was carried out in the presence of normal human serum (NHS), cells rapidly sedimented. The purified cofactor from NHS contained two components of 70 000 and 50 000 molecular weight (MW), as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The latter was recognized by the RRF IgM on immunoblot and its N-terminal sequence indicated that it was beta2-glycoprotein 1 (beta2-GP1), an anionic phospholipid-binding protein. Methanol-treated formalinized SRBC did not support the RR. Thin-layer chromatography of an extract of membranes indicated that the SRBC ligand was a cell-surface phospholipid. Cardiolipin inhibited the RR. These data demonstrate that the RR involves a trimolecular interaction in which IgM, beta2-GP1 and an SRBC phospholipid participate. By analogy with the antiphospholipid antibodies (anti-PL) that occur in autoimmune processes, serum samples from 29 systemic lupus erythematosus patients with high levels of anticardiolipin antibodies were submitted to the RR. A positive RR was obtained for 45% (13 of 29 patients). These results modify the paradigm of the absolute specificity of the RR for leprosy and demonstrate that RRF IgM is a beta2-GP1-dependent anti-PL.