Host inflammatory response to polypropylene implants: insights from a quantitative immunohistochemical and birefringence analysis in a rat subcutaneous model

To describe acute and sub acute aspects of histological and immunohistochemical response to PP implant in a rat subcutaneous model based on objective methods. Thirty rats had a PP mesh subcutaneously implanted and the same dissection on the other side of abdomen but without mesh (sham). The animals were euthanized after 4 and 30 days. Six slides were prepared using the tissue removed: one stained with hematoxylin-eosin (inflammation assessment); one unstained (birefringence evaluation) and four slides for immunohistochemical processing: IL-1 and TNF-α (pro-inflammatory cytokines), MMP-2 (collagen metabolism) and CD-31 (angiogenesis). The area of inflammation, the birefringence index, the area of immunoreactivity and the number of vessels were objectively measured. A larger area of inflammatory reaction was observed in PP compared to sham on the 4th and on the 30th day (p=0.0002). After 4 days, PP presented higher TNF (p=0.0001) immunoreactivity than sham and no differences were observed in MMP-2 (p=0.06) and IL-1 (p=0.08). After 30 days, a reduction of IL-1 (p=0.010) and TNF (p=0.016) for PP and of IL-1 (p=0.010) for sham were observed. Moreover, area of MMP-2 immunoreactivity decreased over time for PP group (p=0.018). Birefringence index and vessel counting showed no differences between PP and sham (p=0.27 and p=0.58, respectively). The implantation of monofilament and macroporous polypropylene in the subcutaneous of rats resulted in increased inflammatory activity and higher TNF production in the early post implant phase. After 30 days, PP has similar cytokines immunoreactivity, vessel density and extracellular matrix organization423585593FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2011/11522-

Importancia da atividade da pseudocolinesterase em pacientes portadores de hepatopatias cronicas

Orientador: Luis Alberto MagnaTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaDoutoradoGenetica e EvoluçãoDoutor em Ciências Biológica

Investigação da atividade e da concentração da anidrase carbonica eritrocitaria em pacientes com quadro clinico sugestivo da doença de Alzheimer

Orientador : Luis Alberto MagnaDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BilogiaResumo: A anidrase carbônica (AC) (EC 4.2.1.1) é uma metaloenzima que contém um átomo de zinco. Sua função de maior importância é a hidratacão do dióxido de carbono e a desidratação do ácido carbônico, sendo o seu substrato natural o dióxido de carbono. Na espécie humana foram reconhecidas até opresente. sete isozimas da anidrase carbônica, as quais diferem entre. si quanto à estrutura molecular propriedades cinéticas, especificidade, pelo substrato. Padrões de inibição, de eficiência catalitica. além da distribuição tecidual. A mais alta concentração da anidrase carbônica é encontrada nos eritrócitos, nos quais se reconhecem três isozimas (CAI, CAII e CAIII). Sabe-se, atualmente. que algumas doenças estão associadas a variações quantitativas e/ou qualitativas das isozimas eritrocitárias da anidrase carbônica. Visto que as pessoas com trissomia do cromossomo 21 (síndrome de Down) mostram aumento significativo da quantidade da CAI eritrocitária, passou a ser de crucial importância investigar se ocorreria fenômeno idêntico nos pacientes com a doença de Alzheimer, pois é notória a coincidência de alguns sinais nessas duas enfermidades,tais como a degeneração granulovacuolar de neurônios, placas senis. enovelados neurofibri1ares, calci;icaç:ões cerebrais e alterôções do metabolismo dos peróxidos. A atividade total da anidrase carbônica eritrocitária e as concentrações de suas isolimas e do zinco plasmático foram investigadas em 10 pacientes com diagnóstico clínico confinante à certeza de doença de Alzheimer e em 10 indivíduos sadios, que se assemelhavam aos pacien tes quanto à idade, sexo e raça (grupo controle). A determinação da atividade total da anidrase carbônica baseou-se na sua inibição por acetazolamida, usando o acetato de alfa-naftila como substrato. Âs concentrações das isozimas foram determinadas por densitometria, após eletro forese, em fitas de acetato de celulose, das soluções parcialmente purificadas. A concentração de zinco plasmático foi obtida por intermédio de espectrofotômetro de absorção atômica com chama. A atividade total da anidrase carbônica dos pacientes não diferiu da observada no grupo controle. Esses dois grupos também não diferiram quanto à distribuição das isozimas eritrocitârias e a concentração de zinco plasmático. Tais resultados conduzem à seguinte conclusão:o padrão da anidrase carbônica eritrocitária dos pacientes co. quadro clínico sugestivo de doença de Alzheimer é diferente daquele encontrado em indivíduos com sindrome de Down, mas não difere daquele apresentado por pessoas normaisAbstract: Carbonic anhydrase, (CA) (EC 4.2.1.1) is a zinc-containing mettalloenzym, wich central function is the hydration of carbon dioxide and the dehydration of carbonic acid, and íts naturalsubstrate is carbon dioxide. So far, in human species only seven isozymes of carbonic Inhydrase have been identified. wich differ from each other concerning their molecular structure, kinect propertíes, specifit_ from substrate, ínhibiton sensitivity and catalytíc in addition to tissue distribuition. The highest concentration of carbonic anhydrase is founded in erythrocytes. in wich three isozymes <CAI, CAII and CAIII) are recognized. As is well know , some diseases are aS5cciated to quantitative or qualitative variations of carbonic anhydrases erythrocytes isozymes. Cocerning that people with Down.s Syndrome (chrom0550me 21 tryssomy) show significative increase of erythrocyte CAI, it seemed extremely Important to investigate if the same occurs in pacients with Alzheimer¿s disease, since there is a prominant coincidence in some of the signs of the two diseases, such a5 granulovacuolar degeneration of neurons, senile plaques, neurofibrillary tang1es, cerebelary ca1cifications and alterations of peroxide metabolism. The whole activity of erythrocytic carbonic anhydrase and its isozymes and plasmatic zinc concentrations were investigated in 10 diseased with cl1inical diagnostic suggesting Alzheimer's disease and 10 healthy.people, with simmilar caracteristics of the diseased as for age, sex and race (control group). The determination of the whole activity of carbonic anhydrase was based on its inhibition by acetazolamide, using alpha-naphtyl acetate as 5ubstrate. Concentration of isozymes was determinated by densitometry, after electrophoresis in cel1u1ose acetate membrane of partial1y purified so1utions. Concentration of plasmatic Zinc was measured by atomic abSoption spectrophotometry (with f1ame).Carbonic anhydrase total activity was similar in diseased group and the control group, as well as both groups did not differed concerning the distribuition of erythrocytic isozymes and plasmatic zinc . These results conduct to the follow conclusion : the role of erythrocytic carbonic anhydrase of the people with clinical diagnostic suggesting Alzheimer's disease is different to that the found in people with Down's syndromeMestradoMestre em Ciências Biológica

Host inflammatory response to polypropylene implants: insights from a quantitative immunohistochemical and birefringence analysis in a rat subcutaneous model

ABSTRACT Objectives To describe acute and sub acute aspects of histological and immunohistochemical response to PP implant in a rat subcutaneous model based on objective methods. Materials and Methods Thirty rats had a PP mesh subcutaneously implanted and the same dissection on the other side of abdomen but without mesh (sham). The animals were euthanized after 4 and 30 days. Six slides were prepared using the tissue removed: one stained with hematoxylin-eosin (inflammation assessment); one unstained (birefringence evaluation) and four slides for immunohistochemical processing: IL-1 and TNF-&#945; (pro-inflammatory cytokines), MMP-2 (collagen metabolism) and CD-31 (angiogenesis). The area of inflammation, the birefringence index, the area of immunoreactivity and the number of vessels were objectively measured. Results A larger area of inflammatory reaction was observed in PP compared to sham on the 4th and on the 30th day (p=0.0002). After 4 days, PP presented higher TNF (p=0.0001) immunoreactivity than sham and no differences were observed in MMP-2 (p=0.06) and IL-1 (p=0.08). After 30 days, a reduction of IL-1 (p=0.010) and TNF (p=0.016) for PP and of IL-1 (p=0.010) for sham were observed. Moreover, area of MMP-2 immunoreactivity decreased over time for PP group (p=0.018). Birefringence index and vessel counting showed no differences between PP and sham (p=0.27 and p=0.58, respectively). Conclusions The implantation of monofilament and macroporous polypropylene in the subcutaneous of rats resulted in increased inflammatory activity and higher TNF production in the early post implant phase. After 30 days, PP has similar cytokines immunoreactivity, vessel density and extracellular matrix organization