Histology and histochemistry of the accessory gland of the female reproductive tract of Rhodnius neglectus Lent, 1954 (Hemiptera: Reduviidae).

Rhodnius neglectus is a wild triatomine, vector of the protozoan Trypanosoma cruzi, which causes Chagas' disease, and feeds on the blood of small mammals, being essential for its growth and reproduction. Accessory glands of the female reproductive tract are important in insect reproduction, but their anatomy and histology in R. neglectus are poorly studied. The aim of this work was to describe the histology and histochemistry of the accessory gland of the female reproductive tract of R. neglectus. The reproductive tract of five females of R. neglectus was dissected and the accessory glands transferred to Zamboni's fixative solution, dehydrated in a crescent series of ethanol, embedded in historesin, sectioned at 2 µm thick, stained with toluidine blue for histological analysis or mercury bromophenol blue for detection of total proteins. The accessory gland R. neglectus is tubular, without branches, opening in the dorsal region of the vagina and differing along its length in proximal and distal regions. In the proximal region, the gland is lined by the cuticle with a layer of columnar cells associated with muscle fibers. In the distal region of the gland, the epithelium has spherical secretory cells with terminal apparatus and conducting canaliculi opening in the lumen through pores in the cuticle. Proteins were identified in the gland lumen, terminal apparatus, nucleus and cytoplasm of secretory cells. The histology of the R. neglectus gland is similar to that found in other species of this genus, but with variations in the shape and size of its distal region.

Evaluation of cell damage and modulation of cytokines TNF-α, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy / Avaliação do dano celular e da modulação das citocinas TNF-α, IL-6 e IL-10 em macrófagos expostos à terapia fotodinâmica mediada pela PpIX

Abstract Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (MΦ), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated MΦ cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal MΦ from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-α, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 10-30%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF-α and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in MΦ.

Resumo Pouco se sabe se a morte celular induzida pela terapia fotodinâmica (PDT) compromete os macrófagos (MΦ), envolvidos nas respostas imunes induzidas pela PDT. Neste estudo, foram avaliados parâmetros de citotoxicidade dos MΦ mediada pela PDT e a produção de citocinas, frente à protoporfirina IX (PpIX). MΦ peritoneais de camundongos BALB/c foram estimulados in vitro com PDT, luz, PpIX ou lipopolissacarídeo (LPS). Após isto, a viabilidade celular (VC), a lipoperoxidação, os níveis de óxido nítrico (NO), de DNA degradado, de TNF-α, IL-6 e IL-10 foram avaliados. A exposição curta à PDT reduziu a VC em 10-30%. Os níveis de NO e de DNA degradado duplicaram, sem aumento da lipoperoxidação. Houve aumento de TNF-α e IL-10, sendo maior na presença de LPS. Já a produção de IL-6 reduziu em dez vezes. A PDT induz estresse celular, gera radicais NO e causa dano ao DNA, tornando o microambiente citotóxico. Ainda, modula citocinas pró e anti-inflamatórias em MΦ.

Evaluation of cell damage and modulation of cytokines TNF-α, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy.

Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (MΦ), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated MΦ cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal MΦ from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-α, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 10-30%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF-α and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in MΦ.

Morphology of the spermatheca of Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) / Morfologia da espermateca de Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859)

Abstract Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) is a potential vector of Chagas's disease and the comprehension of its reproductive biology is an important tool to control this insect. In the reproductive tract of female insects, the spermatheca plays a crucial role storing male spermatozoa after mating. Whithin insects the spermatheca shows a wide morphological diversity and the analysis of this characteristic can contribute to understand the reproductive biology of the species. This study describes the histology and histochemistry of the spermatheca of T. lecticularia. Females have a pair of elongated spermathecal reservoirs without associated accessory gland. The reservoir opens into the common oviduct via a narrow muscular duct. The reservoir epithelium has single layer of columnar secretory cells. The control of the release of spermatozoa from the spermatheca occurs via the muscular duct. The anatomical features of the spermatheca of T. lecticularia resemble those described of other Reduviidae. However, the histological and histochemical features of spermatheca observed in T. lecticularia were important to explain the maintenance of the viability of the spermatozoa stored.

Resumo Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) é um potencial vetor da doença de Chagas e a compreensão de sua biologia reprodutiva é um importante fator para seu controle populacional. No aparelho reprodutor feminino dos insetos, a espermateca desempenha a importante funcão de armazenar os espermatozoides após cópula. Nos insetos, a espermateca apresenta uma ampla diversidade morfológica e a análise destas características pode contribuir com o entendimento da biologia reprodutiva das espécies. Este estudo descreve histológica e histoquimicamente a espermateca de T. lecticularia. As fêmeas tem um par de espermatecas alongadas sem glândulas acessórias associadas. O reservatório conecta-se ao oviduto comum através de um ducto muscular curto que controla a liberação dos espermatozoides. O epitélio do reservatório possui uma camada de células secretoras colunares. As características anatômicas da espermateca de T. lecticularia são semelhantes às encontradas em outros Reduviidae. Entretanto, as características histológicas e histoquímicas observadas na espermateca são importantes para explicar a manutenção da viabilidade dos espermatozoides armazenados.

Morphology of the spermatheca of Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859).

Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) is a potential vector of Chagas's disease and the comprehension of its reproductive biology is an important tool to control this insect. In the reproductive tract of female insects, the spermatheca plays a crucial role storing male spermatozoa after mating. Whithin insects the spermatheca shows a wide morphological diversity and the analysis of this characteristic can contribute to understand the reproductive biology of the species. This study describes the histology and histochemistry of the spermatheca of T. lecticularia. Females have a pair of elongated spermathecal reservoirs without associated accessory gland. The reservoir opens into the common oviduct via a narrow muscular duct. The reservoir epithelium has single layer of columnar secretory cells. The control of the release of spermatozoa from the spermatheca occurs via the muscular duct. The anatomical features of the spermatheca of T. lecticularia resemble those described of other Reduviidae. However, the histological and histochemical features of spermatheca observed in T. lecticularia were important to explain the maintenance of the viability of the spermatozoa stored.

Evaluation of cell damage and modulation of cytokines TNF-, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy

Abstract Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (M), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated M cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal M from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 1030%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF- and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in M.

Resumo Pouco se sabe se a morte celular induzida pela terapia fotodinâmica (PDT) compromete os macrófagos (M), envolvidos nas respostas imunes induzidas pela PDT. Neste estudo, foram avaliados parâmetros de citotoxicidade dos M mediada pela PDT e a produção de citocinas, frente à protoporfirina IX (PpIX). M peritoneais de camundongos BALB/c foram estimulados in vitro com PDT, luz, PpIX ou lipopolissacarídeo (LPS). Após isto, a viabilidade celular (VC), a lipoperoxidação, os níveis de óxido nítrico (NO), de DNA degradado, de TNF-, IL-6 e IL-10 foram avaliados. A exposição curta à PDT reduziu a VC em 10-30%. Os níveis de NO e de DNA degradado duplicaram, sem aumento da lipoperoxidação. Houve aumento de TNF- e IL-10, sendo maior na presença de LPS. Já a produção de IL-6 reduziu em dez vezes. A PDT induz estresse celular, gera radicais NO e causa dano ao DNA, tornando o microambiente citotóxico. Ainda, modula citocinas pró e anti-inflamatórias em M.