Identification of carboxymethyl (CM)-binding proteins derived from Lolium multiflorum pollen extract and antibody reactivity in Brazilian allergic patients

Lolium multiflorum grass is the major pollen allergen source in the southern region of Brazil, but most of its allergens remain poorly characterized. The aim of this study was to investigate antibody reactivity to L. multiflorum crude and carboxymethyl-ligand extracts in allergic patients and healthy individuals. Ion exchange carboxymethyl (CM) chromatography (CM-Sepharose) was used to isolate proteins (S2) from L. multiflorum crude extract (S1), which were assessed by SDS-PAGE. S1- and S2-specific IgE and IgG4 levels were measured by ELISA using sera from 55 atopic and 16 non-atopic subjects. Reactive polypeptide bands in S1 and S2 were detected by immunoblotting, and the most prominent bands in S2 were analyzed by mass spectrometry (MS-MS). Similar IgE and IgG4 levels were observed to both S1 (IgE median absorbance: 1.22; IgG4 median absorbance: 0.68) and S2 (IgE median absorbance: 1.26; IgG4 median absorbance: 0.85) in atopic subjects. S1 and S2 had positive correlations for IgE and IgG4 (IgE: r=0.9567; IgG4: r=0.9229; P<0.0001) levels. Homology between S1 and S2 was confirmed by IgE (84%) and IgG4 (83%) inhibition. Immunoblotting revealed that the 29-32 kDa band was recognized by 100% of atopic subjects in both S1 and S2. MS-MS analysis identified similarity profile to groups 1 and 5 grass allergens. This study revealed that carboxymethyl-ligand fraction played an important role for pollen allergy diagnosis by containing clinically relevant allergens and constituted a promising candidate for allergen-specific immunotherapy.

Single-walled carbon nanotubes functionalized with sodium hyaluronate enhance bone mineralization

The aim of this study was to evaluate the effects of sodium hyaluronate (HY), single-walled carbon nanotubes (SWCNTs) and HY-functionalized SWCNTs (HY-SWCNTs) on the behavior of primary osteoblasts, as well as to investigate the deposition of inorganic crystals on titanium surfaces coated with these biocomposites. Primary osteoblasts were obtained from the calvarial bones of male newborn Wistar rats (5 rats for each cell extraction). We assessed cell viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay and by double-staining with propidium iodide and Hoechst. We also assessed the formation of mineralized bone nodules by von Kossa staining, the mRNA expression of bone repair proteins, and the deposition of inorganic crystals on titanium surfaces coated with HY, SWCNTs, or HY-SWCNTs. The results showed that treatment with these biocomposites did not alter the viability of primary osteoblasts. Furthermore, deposition of mineralized bone nodules was significantly increased by cells treated with HY and HY-SWCNTs. This can be partly explained by an increase in the mRNA expression of type I and III collagen, osteocalcin, and bone morphogenetic proteins 2 and 4. Additionally, the titanium surface treated with HY-SWCNTs showed a significant increase in the deposition of inorganic crystals. Thus, our data indicate that HY, SWCNTs, and HY-SWCNTs are potentially useful for the development of new strategies for bone tissue engineering.

Aplicações da cultura de tecidos em plantas medicinais / Applications of tissue culture in medicinal plants

Esta revisão tem por objetivo levantar dados de literatura sobre o histórico e a situação atual das técnicas de cultura de tecidos em plantas medicinais. Para tanto, foi realizada uma revisão de publicações do período de 1976 a 2009. A cultura de tecidos é muito utilizada em pesquisas envolvendo plantas medicinais, com destaque para a técnica de micropropagação. A aplicação das técnicas de cultura de tecidos em plantas medicinais tem como perspectivas a obtenção de germoplasma competitivo e adaptado a diversos métodos de cultivo, escolha de novas espécies que servirão como fonte de compostos biologicamente ativos e aprimoramento da produção de fitofármacos, a fim de assegurar exploração sustentável destas espécies.

The aim of this literature review is to conduct a survey concerning the history and current situation of tissue culture techniques in medicinal plants. Therefore, a review was done considering the period from 1976 to 2009. Tissue culture is widely applied in medicinal plants researches, especially micropropagation. The perspectives of tissue culture techniques in medicinal plants are related to the development of competitive germoplasm adapted to diverse methods of cultivation, the election of new species that will serve as source of biological active composts, and the improvement of phytochemicals production, in order to assure sustainable exploration of these species.

Concentrações de BAP sobre a proliferação in vitro de brotos de Lippia alba [(Mill. )N. E. Brown] / BAP concentrations on in vitro proliferation of Lippia alba [(Mill. ) N. E. Brown] shoots

Lippia alba é uma planta de origem brasileira, pertencente à família Verbenaceae. É conhecida por combater a insônia e a asma, além de possuir ação anticonvulsivante e antifúngica. A micropropagação tem sido utilizada para multiplicação de várias espécies com propriedades medicinais, e o BAP é a citocinina sintética mais utilizada para a multiplicação in vitro. O objetivo deste trabalho foi estabelecer a concentração adequada de BAP na multiplicação in vitro de L. alba. Explantes constituídos de segmentos nodais provenientes de plântulas já estabelecidas in vitro com aproximadamente 0,5 cm foram inoculados em meio MS, suplementado com 0,0; 0,5; 1,0 e 1,5 mg L-1 de BAP em adição de 30 g L-1 de sacarose. A utilização de 1,5 mg L-1 de BAP promove a multiplicação in vitro de L. alba. A ausência deste regulador propicia maior número de folhas e o uso na concentração de 0,5 mg L-1 aumentou as massas fresca e seca de parte aérea nesta espécie.

Lippia alba is a Brazilian plant, belonging to the Verbenaceae family. It is known to combat insomnia and asthma, also has antifungal and anticonvulsant actions. Micropropagation has been used for propagation of various species with medicinal properties, and BAP is the most widely used synthetic cytokinin for in vitro multiplication. The aim of this study was to determine the appropriate concentration of BAP on in vitro multiplication of L. alba. Explants consisting of nodal segments from seedlings already established in vitro with approximately 0.5 cm were inoculated on MS medium supplemented with 0; 0.5; 1.0 and 1.5 mg L-1 of BAP and 30 g L-1 of sucrose. The use of 1.5 mg L-1 of BAP promotes the in vitro multiplication of L. alba. The absence of this regulator provides a greater number of leaves and its use at the concentration of 0.5 mg L-1 increased shoots' fresh and dry weights in this specie.

Citocininas na multiplicação in vitro de hortelã-pimenta (Mentha x Piperita L. ) / Cytokinins on in vitro multiplication of peppermint (Mentha x Piperita L. )

O hortelã-pimenta é um hibrido triplo, Mentha x piperita, utilizado de forma medicinal no tratamento de náuseas, cólicas gastrointestinais, flatulências, cálculos biliares, icterícia, ansiedade, expectoração e expulsão de vermes intestinais. Na micropropagação, o BAP (6-benzilaminopurina) e KIN (cinetina) têm sido as fontes de citocininas mais empregadas. O objetivo deste trabalho foi avaliar o efeito de reguladores vegetais na multiplicação in vitro de hortelã-pimenta. Explantes constituídos de segmentos nodais provenientes de plântulas já estabelecidas in vitro com aproximadamente 0,5 cm foram inoculados em meio MS, suplementado com diferentes associações de BAP e KIN, em adição de 30 g L-1 de sacarose. As concentrações de citocininas utilizadas foram 0,0; 1,0 e 2,0 mg L-1 de BAP e 0,0; 0,5 e 1,0 mg L-1 de KIN. O delineamento experimental foi o inteiramente casualizado, com nove tratamentos consistindo de cinco frascos cada um, sendo que cada frasco continha quatro explantes. A utilização de 2,0 mg L-1 de BAP promove a multiplicação in vitro de hortelã-pimenta, porém, diminui o índice de sobrevivência. O uso de citocininas aumenta a massa fresca e seca dos explantes e a ausência destes reguladores propicia o alongamento nesta espécie.

Peppermint is used for medical treatments of nausea, gastrointestinal cramps, flatulence, gallstones, jaundice, anxiety, sputum and expulsion of intestinal worms. In micropropagation, BAP (6-benzylaminopurine) and KIN (kinetin) have been the most widely used sources of cytokinins. The aim of this study was to evaluate the effect of plant growth regulators in multiplication of peppermint. Explants consisting of nodal segments from seedlings already established in vitro with approximately 0,5 cm were inoculated on MS medium supplemented with different combinations of BAP and KIN and 30 g L-1 of sucrose. The concentrations of cytokinins used were 0,0; 1,0 and 2,0 mg L-1 of BAP and 0,0; 0,5 and 1,0 mg L-1 of KIN. The experiment was set up in a completely randomized design with nine treatments consisting of five bottles each, and each vial contained four explants. The use of 2,0 mg L-1 of BAP promotes the in vitro multiplication of peppermint, but decreases the survival rate. Cytokinins increase fresh and dry weight of explants and the absence of these regulators provides the elongation in this specie.

Familial osteopetrosis in Agouti paca: report of nine cases / Osteopetrose familiar em Agouti paca: relato de nove casos

Nine cases of familial osteopetrosis were studied in Agouti paca rodents maintained in captivity. Animals were distributed in three groups depending on the severity of their skeletal lesions. Based upon clinical, radiological, and microscopic findings, it was concluded that one animal had level I lesions, three animals had level II lesions, and five animals had level III osteopetrosis and osteonecrosis. Throughout the entire axial and appendicular skeleton, there was an increased amount of both trabecular and cortical bone tissue. All analyzed bones showed thickened cortex and reduced medullary canals. Bone trabeculae were thick and confluent. Cortex showed a narrowing of Haversian canals. Numerous cementing lines resulted in typical mosaic patterns. Osteocytes were pycnotic. Osteonecrosis was characterized by the disappearance of osteocytes and bone matrix decomposition.

Descreveram-se nove casos de osteopetrose familiar em Agouti paca mantidas em cativeiro. Os animais foram distribuídos em três grupos de acordo com a gravidade das lesões do esqueleto. Com base nos exames clínico, radiológico e microscópico, foi concluído que um animal apresentou lesões de nível I, três animais tiveram lesões de nível II e cinco animais tiveram osteopetrose de nível III. Por todo o esqueleto axial e apendicular, a quantidade de osso trabecular e osteônico estava aumentada. Todos os ossos analisados mostraram córtex espesso e canais medulares reduzidos. As trabéculas ósseas eram espessas e confluentes. No córtex, verificou-se um estreitamento de canais de Havers. Numerosas linhas de cimentação resultaram em um padrão de mosaico típico. Osteócitos estavam picnóticos e a osteonecrose foi caracterizada pela morte dos osteócitos, com desintegração da matriz óssea.

Responses of IgE, IgG1, and IgG4 to concanavalin A-binding Blomia tropicalis antigens in allergic patients

Blomia tropicalis (Bt) and Dermatophagoides pteronyssinus (Dp) are the prevalent house dust mites in tropical countries and are associated with allergic diseases. Glycosylated antigens are highly immunogenic and involved in different pathologies. We evaluated the presence of IgE, IgG1, and IgG4 to concanavalin A-binding antigens (Bt-Con-A) isolated from Bt-total extract in sera of allergic and non-allergic subjects. Bt-total and Bt-Con-A extracts were evaluated by SDS-PAGE and ELISA for reacting with IgE, IgG1, and IgG4 in sera of 121 patients with allergic rhinitis and 36 non-allergic individuals. All subjects were skin prick tested with Bt-total extract and inhibition tests were performed for IgE, IgG1, and IgG4 using both extracts (Bt-total and Bt-Con-A). Skin prick test showed that 58 percent of the patients were sensitized to Bt (Bt+), with 52 percent reactive to both mites (Bt and Dp) and 6 percent to Bt only. A broad spectrum of proteins (14-152 kDa) was visualized in Bt-total and components >27 kDa for the Bt-Con-A extract. ELISA showed a similar profile of IgE, IgG1 and IgG4 levels in response to Bt-total and Bt-Con-A extracts in different groups, although Bt+ patients showed a lower IgG4 reactivity to Bt-Con-A extract. Specific IgG1 levels were higher in Bt+ patients than in control subjects, and IgG4 levels showed no significant difference among groups. ELISA inhibition showed a partial IgE and total IgG1 and IgG4 cross-reactivity with Dp extract for Bt-total and Bt-Con-A extracts. We conclude that Con-A-binding components isolated from Bt constitute major allergens and are involved in both allergen sensitization (IgE response) and homeostasis maintenance (IgG1 and IgG4 responses).

Competência social de psicóticos: validaçäo social de habilidades específicas / Social skills of psychiatric patients: socialvalidation of specific skills

Esta pesquisa identifica as dificuldades de interaçäo social de pacientes psiquiátricos, em termos de medidas moleculares da competência social, referentes aos componentes verbais, näo-verbais, paralingüísticos, de expressividade e da habilidade de soluçäo de problemas dos sujeitos em quatro situaçöes cotidianas. Foi utilizada a estratégia de validaçäo social, na qual 35 pacientes psiquiátricos foram comparados a um grupo de referência de sua própria comunidade. Dois juízes previamente treinados observaram e cotaram a presença e ausência de comportamento específicos para cada um dos componentes descritos acima, utilizando uma grade de observaçäo sistemática do comportamento (EEHS). Os resultados mostram que as dificuldades dos pacientes säo mais acentuadas com relaçäo ao componente verbal e desoluçäo de problemas, mas envolvem também os componentes paralingüísticos, näo verbal e de expressividade. As dificuldades observadas, dentro de cada um dos componentes acima, se referem a alguns comportamentos particulares e näo a outros, identificando a especificifdade do déficit apresentado por esta clientela e portanto a necessidade de intervençöes clínicas específicas. Em alguns aspectos da competência social, o grupo de referência apresenta dificuldades semelhantes ao do grupo clínico. Os resultados säo discutidos em termos da importância da estratégia de validaçäo social para a escolha de objetivos socialmente pertinenetes em sua comunidade