iNOS-derived Nitric Oxide Modulates Infection-stimulated Bone Loss

Nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) plays an important role in host defense, as well as in inflammation-induced tissue lesions. Here we evaluated the role of NO in bone loss in bacterial infection-induced apical periodontitis by using iNOS-deficient mice (iNOS(-/-)). The iNOS(-/-) mice developed greater inflammatory cell recruitment and osteolytic lesions than WT mice. Moreover, tartrate-resistant acid-phosphatase-positive (TRAP(+)) osteoclasts were significantly more numerous in iNOS-/- mice. Furthermore, the increased bone resorption in iNOS(-/-) mice also correlated with the increased expression of receptor activator NF-kappaB (RANK), stromal-cell-derived factor-1 alpha (SDF-1 alpha/CXCL12), and reduced expression of osteoprotegerin (OPG). These results show that NO deficiency was associated with an imbalance of bone-resorption-modulating factors, leading to severe infection-stimulated bone loss.

Noninvasive Evaluation of Oral Lesions Using Depth-sensitive Optical Spectroscopy

BACKGROUND: Optical spectroscopy is a noninvasive technique with potential applications for diagnosis of oral dysplasia and early cancer. In this study, we evaluated the diagnostic performance of a depth-sensitive optical spectroscopy (DSOS) system for distinguishing dysplasia and carcinoma from non-neoplastic oral mucosa. METHODS: Patients with oral lesions and volunteers without any oral abnormalities were recruited to participate. Autofluorescence and diffuse reflectance spectra of selected oral sites were measured using the DSOS system. A total of 424 oral sites in 124 subjects were measured and analyzed, including 154 sites in 60 patients with oral lesions and 270 sites in 64 normal volunteers. Measured optical spectra were used to develop computer-based algorithms to identify the presence of dysplasia or cancer. Sensitivity and specificity were calculated using a gold standard of histopathology for patient sites and clinical impression for normal volunteer sites. RESULTS: Differences in oral spectra were observed in: (1) neoplastic versus nonneoplastic sites, (2) keratinized versus nonkeratinized tissue, and (3) shallow versus deep depths within oral tissue. Algorithms based on spectra from 310 nonkeratinized anatomic sites (buccal, tongue, floor of mouth, and lip) yielded an area under the receiver operating characteristic curve of 0.96 in the training set and 0.93 in the validation set. CONCLUSIONS: The ability to selectively target epithelial and shallow stromal depth regions appeared to be diagnostically useful. For nonkeratinized oral sites, the sensitivity and specificity of this objective diagnostic technique were comparable to that of clinical diagnosis by expert observers. Thus, DSOS has potential to augment oral cancer screening efforts in community settings. Cancer 2009;115:1669-79. (C) 2009 American Cancer Society.

Molecular characterization of Mycobacterium massiliense and Mycobacterium bolletii in isolates collected from outbreaks of infections after laparoscopic surgeries and cosmetic procedures

An outbreak of infections affecting 311 patients who had undergone different invasive procedures occurred in 2004 and 2005 in the city of Belem, in the northern region of Brazil. Sixty-seven isolates were studied; 58 were from patients who had undergone laparoscopic surgeries, 1 was from a patient with a postinjection abscess, and 8 were from patients who had undergone mesotherapy. All isolates were rapidly growing nonpigmented mycobacteria and presented a pattern by PCR-restriction enzyme analysis of the hsp65 gene with BstEII of bands of 235 and 210 bp and with HaeIII of bands of 200, 70, 60, and 50 bp, which is common to Mycobacterium abscessus type 2, Mycobacterium bolletii, and Mycobacterium massiliense. hsp65 and. rpoB gene sequencing of a subset of 20 isolates was used to discriminate between these three species. hsp65 and rpoB sequences chosen at random from 11 of the 58 isolates from surgical patients and the postinjection abscess isolate presented the highest degrees of similarity with the corresponding sequences of M. massiliense. In the same way, the eight mesotherapy isolates were identified as M. bolletii. Molecular typing by pulsed-field gel electrophoresis (PFGE) grouped all 58 surgical isolates, while the mesotherapy isolates presented three different PFGE patterns and the postinjection abscess isolate showed a unique PFGE pattern. In conclusion, molecular techniques for identification and typing were essential for the discrimination of two concomitant outbreaks and one case, the postinjection abscess, not related to either outbreak all of which were originally attributed to a single strain of M. abscessus.

Behandlingsmetoder och amningsförekomst vid bröstböld

Bakgrund: Amningens fördelar för mor, barn och samhälle är välkända. En komplikation vid amning är bröstböld. Medvetenhet om fördelar och nackdelar med olika behandlingsmetoder vid bröstböld och deras inverkan på fortsatt amning är viktig. Syftet: Att studera behandlingsmetoder och amningsförekomst vid bröstböld. Metod: Retrospektiv design från en journalgranskning genomförd på Södersjukhuset mellan januari 2006 och september 2008. Statistiska analyser genomfördes med Chi-två test, Independent samples T-test samt Fishers exakta test. Resultat: Totalt 139 kvinnor med bröstböld behandlades med nålpunktions-, pigtail- eller dränagebehandling, eller en kombination av dessa. Sextiotre procent pigtailbehandlades enbart och dessa behövde i mindre omfattning sjukhusinläggning jämfört med ej enbart pigtailbehandlade, samt resulterade i större andel av ett till tre sjukhusbesök. Enbart dränagebehandling och flera metoder resulterade i större andel sjukhusinläggningar än ej enbart dränagebehandling och en metod. Amningsförekomsten sjönk från 96 procent före behandling till 69 procent en månad efteråt. Konklusion: Enbart pigtailbehandling var den vanligaste metoden och resulterade i färre sjukhusinläggningar och sjukhusbesök. Förekomsten av amning hade sjunkit efter behandling av bröstböld

Avaliação do erro das coordenadas de localização estereotáctica guiada por tomografia computadorizada do encéfalo

São analisados 106 pacientes submetidos a localização estereotáctica. Os procedimentos variaram de biópsias cerebrais, orientação de craniotomias, colocação de cateter em cavidade tumoral, drenagem de hematoma intracerebral e drenagem de abscesso cerebral. As orientações de craniotomias foram para MAVs, tumores e processos inflamatórios, em 21 pacientes. As biópsias cerebrais estereotácticas para diagnóstico anatomopatológico apresentaram um índice de positividade de 87,50 % com complicações em 1,20 %, em 82 casos. São analisadas estatisticamente as variáveis como: idade, sexo, procedimento realizado, diagnóstico anatomopatológico e volume das lesões. É discutida a imprecisão na aquisição e cálculo das coordenadas estereotácticas com a TC do encéfalo e verificada a precisão do método estereotomográfico com a utilização de um phanton. O maior erro das coordenadas foi de 6,8 mm.

Abscesso pulmonar de aspiração : dados diagnósticos e resultados terapêuticos em 241 casos

O abscesso de pulmão continua sendo hoje, em plena era dos antibióticos, um importante problema médico. O presente trabalho se propõe mostrar aspectos diagnósticos e terapêuticos da doença em uma série de pacientes coletada nos últimos 34 anos em um hospital universitário especializado em doenças pulmonares. No período de 1968 a 2002 foram reunidos e estudados 241 casos de abscesso pulmonar de aspiração - 199 em homens e 42 em mulheres, com média de idade de 41,3 anos. Em 69,0% dos pacientes esteve presente a ingestão de álcool e em 64,0% o hábito tabágico. Tosse, expectoração, febre e comprometimento do estado geral foram os achados clínicos mais freqüentes, encontrados na quase totalidade dos casos; 62,5% tinham dor torácica e 30,0% hipocratismo digital. Verificaram-se dentes em mau estado de conservação em 81,7 % dos pacientes, episódio de perda de consciência em 78,0% e presença de odor fétido em 66,0%. Em 85,5% das vezes as lesões localizaram-se em segmento posterior de lobo superior ou segmento superior de lobo inferior, 97,1% delas unilaterais, ocorrendo com igual freqüência tanto no pulmão direito como no esquerdo. A maioria das lesões (66,0%) mediram entre 4,0 e 8,0 cm de diâmetro. Em 25 pacientes (10,4%) houve a associação de empiema pleural. Flora mista, indicativa da presença de germes anaeróbios, foi identificada em secreções broncopulmonares ou pleurais em 172 pacientes (71,4 %). Estreptococos e Gram negativos aeróbios foram também algumas vezes encontrados. Todos os pacientes foram inicialmente tratados com antibióticos (penicilina em 78,0% das vezes) e submetidos a sessões de drenagem postural. Em 51 (21,2%) acabou sendo necessário algum procedimento cirúrgico (24 drenagens de empiema, 21 ressecções pulmonares e 6 pneumostomias). Cura foi obtida em 231 pacientes (95,8%) e 10 (4,2%) foram ao óbito, estes em geral com grandes abscessos, três dos quais também com empiema.

Abscesso esplênico

Splenic abscess is a rare disease usually associated with immunosuppressed states. Its diagnosis may be difficult due to non-specific symptoms . The authors report a case of a healthy woman, 42 years old, bearer of splenic and renal abscesses , treated with antibiotics and splenectomy. A review of the literature is presented with emphasis on the diagnosis and treatment

Análise da estabilidade genética de células-tronco mesenquimais humanas

Human multipotent mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, have become an important and attractive therapeutic tool since they are easily isolated and cultured, have in vitro expansion potential, substantial plasticity and secrete bioactive molecules that exert trophic effects. The human umbilical cord as a cell source for cell therapy will help to avoid several ethical, political, religious and technical issues. One of the main issues with SC lines from different sources, mainly those of embryonic origin, is the possibility of chromosomal alterations and genomic instability during in vitro expansion. Cells isolated from one umbilical cord exhibited a rare balanced paracentric inversion, likely a cytogenetic constitutional alteration, karyotype: 46,XY,inv(3)(p13p25~26). Important genes related to cancer predisposition and others involved in DNA repair are located in 3p25~26. Titanium is an excellent biomaterial for bone-implant integration; however, the use can result in the generation of particulate debris that can accumulate in the tissues adjacent to the prosthesis, in the local bone marrow, in the lymph nodes, liver and spleen. Subsequently may elicit important biological responses that aren´t well studied. In this work, we have studied the genetic stability of MSC isolated from the umbilical cord vein during in vitro expansion, after the cryopreservation, and under different concentrations and time of exposition to titanium microparticles. Cells were isolated, in vitro expanded, demonstrated capacity for osteogenic, adipogenic and chondrogenic differentiation and were evaluated using flow cytometry, so they met the minimum requirements for characterization as MSCs. The cells were expanded under different concentrations and time of exposition to titanium microparticles. The genetic stability of MSCs was assessed by cytogenetic analysis, fluorescence in situ hybridization (FISH) and analysis of micronucleus and other nuclear alterations (CBMN). The cells were able to internalize the titanium microparticles, but MSCs preserve their morphology, differentiation capacity and surface marker expression profiles. Furthermore, there was an increase in the genomic instability after long time of in vitro expansion, and this instability was greater when cells were exposed to high doses of titanium microparticles that induced oxidative stress. It is necessary always assess the risks/ benefits of using titanium in tissue therapy involving MSCs, considering the biosafety of the use of bone regeneration using titanium and MSCs. Even without using titanium, it is important that the therapeutic use of such cells is based on analyzes that ensure quality, security and cellular stability, with the standardization of quality control programs appropriate. In conclusion, it is suggested that cytogenetic analysis, FISH analysis and the micronucleus and other nuclear alterations are carried out in CTMH before implanting in a patient

Tratamento de fístulas enterocutâneas através de sistema a vácuo com alta pressão e dieta oral normal

Enterocutaneous fistulas are associated with prolonged hospital stay, high morbidity/mortality, and increase in hospital costs. This study aims to describe the use of a vacuum system and normal oral diet in dealing with this problem. Methods: Seventy-four consecutive patients with recent and defined external postoperative fistulas were analyzed. Abdominal imaging was used to exclude abscess and distal obstruction. The fistula tract was sealed with Foley catheter, connected to a negative pressure flask, changed daily for 5, 10 or 15 days, as necessary. Normal oral diet was permitted. Results: No patient died. Serum albumin and transferrin showed significantly higher levels at the end of treatment than at the beginning. The moderate and low-output fistulas had the best results (97% closed). Forty-eight (65%) fistulas closed after five days, 16(22%) after 10 days and 4(5%) after 15 days. Treatment failed in 6(8%) patients, who subsequently underwent surgery. Only one patient with low-output did not close her fistula. The cost of the treatment was US$ 41.75/day and it was considered cost effective. Conclusions: The vacuum system demonstrated good results in the treatment of fistulas. It included simplicity, low cost, short hospital stay, absence of skin breakdown, normal eating, good nutrition and activity patterns

Avaliação histológica da próstata de cães adultos sexualmente intactos

Avaliou-se histologicamente a próstata de 30 cães adultos e idosos sexualmente intactos que apresentavam ou não sintomatologia clínica de doença prostática, e verificou-se a incidência de possíveis alterações da glândula. Dentre as alterações encontradas, a hiperplasia prostática benigna constituiu o diagnóstico mais comum, 85,6% (n=24), seguida por prostatite crônica, 64,3% (n=18), displasia do epitélio glandular, 42,8% (n=12), atrofia do epitélio glandular, 39,3% (n=11), infiltrado inflamatório focal, 25% (n=7), dilatação glandular focal, 21,4% (n=6), prostatite aguda, 7,1% (n=2), metaplasia escamosa, 3,6%, (n=1), metástase de neoplasia sistêmica, 3,6% (n=1) e abscesso prostático, 3,6% (n=1). Como em muitos casos os cães são assintomáticos, ressalta-se a importância da realização rotineira de exames clínicos específicos, como o toque retal e a ultrassonografia, para o diagnóstico precoce e o tratamento das afecções prostáticas.

Viabilidade celular da fração mononuclear da medula óssea e fração vascular estromal do tecido adiposo de equinos após o processo de congelamento e descongelamento

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nandrolone Decanoate and Physical Effort: Histological and Morphometrical Assessment in Adult Rat Uterus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental dermatophytosis in hamsters inoculated with Trichophyton mentagrophytes in the cheek pouch

Esse estudo apresenta os resultados obtidos quando da inoculação de Trychophyton mentagrophytes na bolsa jugal do hamster, local imunologicamente privilegiado. Foram utilizados 42 animais: 21 inoculados com 10(6) fungos na bolsa jugal (grupo 1) e, 21 inicialmente inoculados com 10(6) fungos no coxim plantar e, 15 dias após, na bolsa jugal com a mesma quantidade fúngica (grupo 2). Os animais foram sacrificados às 20 h, 3, 7, 14, 30, 60 e 120 dias; foram coletadas amostras da bolsa jugal inoculada, e das patas submetidas ao teste do coxim plantar (TCP). Independente do grupo e do tempo de evolução da infecção, os animais não desenvolveram hipersensibilidade tardia avaliada através do TCP. A pré-inoculação de fungos no coxim plantar não alterou a morfologia das lesões induzidas na bolsa jugal. Assim, nos animais do grupo 1 e grupo 2, a introdução do fungo na bolsa jugal, resultou em lesão focal, constituída por infiltrado inflamatório agudo estéril, com formação de abscesso, que evoluiu para reação macrofágica e, posteriormente, para a resolução mesmo na ausência de resposta imune detectável pelo TCP. Nossos resultados indicam que, apesar do importante papel da resposta imune na regressão espontânea da dermatofitose, outros fatores são, também, parte integral da defesa contra esta infecção fúngica.

Histological evaluation of the lesion induced by inoculation of Leishmania mexicana in the cheek pouch of the hamster

No presente estudo, investigamos o papel da resposta imune na morfologia do granuloma leishmaniótico induzido na bolsa jugal do hamster, um local imunologicamente privilegiado, após inoculação de 3x10(5) Leishmania mexicana. Os animais foram avaliados histológica e imunologicamente até os 120 dias da inoculação. Independente da época do sacrifício, os animais foram sempre não reatores ao teste do coxim plantar. Histologicamente, a inoculação de Leishmania mexicana na bolsa jugal resultou na formação de abcesso que evoluiu para reação granulomatosa rica em formas amastigotas e, posteriormente, para resolução. Esses resultados sugerem que o desenvolvimento da resposta imune não é preponderante no controle da infecção induzida pela Leishmania mexicana inoculada subcutaneamente na bolsa jugal do hamster. Sugerem ainda que os macrófagos que compõe os granulomas leishmanióticos são capazes de eliminar esse parasita, independente da presença de resposta imune avaliável pelo teste do coxim plantar.

Avaliação imuno-histoquímica da BMP-2, BMP-4 e seus receptores (BMPRIA e BMPRII) em ameloblastomas e tumor odontogênico adenomatóide

BMPs are components superfamily ligands transformation growth fator-β (TGF-β) secreted into the extracellular environment, with mechanisms of intercellular communication through specific ligands and receptors in various target cells, being recognized for its influence in osteogenic induction, also play an important role in tissue homeostasis, cell proliferation, differentiation control , in addition to being present in the development of various malignancies. The aim of this study was to compare the immunohistochemical expression of BMP-2, BMP-4 and its receptors BMPRIA and BMPRII in cases of ameloblastoma and adenomatoid odontogenic tumor. The sample consisted of 20 cases of solid ameloblastoma (SA), 10 cases of ameloblastoma unicystic (UA) and 16 cases of adenomatoid odontogenic tumor (AOT). The expression of BMPs and their receptors was evaluated in the parenchyma and stroma of lesions, establishing the percentage of immunopositive cells (0 - negative; 1-1 % to 10 % of cells positive; 2 - 11% to 25% of positive cells; 3 - 26% to 50% of cells positive; 4 - 51% to 75 % of positive cells; 5 - more than 75% positive cells). Analysis of the expression of BMP-2 revealed no statistically significant differences in parenchymal (p = 0.925) and stromal component (p = 0.345) between the groups, as well as BMP-4 (p = 0.873 / p = 0.131). In the epithelial component, SA and AOT had a higher frequency of score 5. In turn, all cases of UA were classified as score 5. The analysis of the stromal component showed no statistically significant difference between groups with respect to median scores BMPRIA positivity (p = 0.768) and BMPRII (p = 0.779). In the epithelial component of SA and UA, no statistically significant correlations between imunoexpression proteins analyzed were observed. In turn, the group of AOT, statistically significant positive correlations between the scores of expression of all studied proteins were found. In the stromal component, statistically significant positive correlations were found only in the SA group in BMP -4 and BMPRII (r = 0.476; p = .034), in the UA in BMP-4 and BMPRIA (r = 0.709; p = 0.022). The results of this study suggest that the BMPs and their receptors are involved in the development process odontogenic tumors. BMP-4, in turn, besides being present in odontogenic tumors have the capacity to form mineralized material.