N-acetyl Glucosamine Distribution and Mitochondrial Activity of Tumor Cell Exposed to Photodynamic Therapy.

The use of lectins can play an important role for tracking modification on cell surface components, since lectins can be easily complexed with radioisotopes, biotin or fluorescein, facilitating the evaluation of carbohydrates distribution in the cell and mitochondrial activity. The aim of this study was to evaluate photodynamic therapy effects on indirect distribution of N-acetyl-glucosamine terminal glycoproteins, in human laryngeal carcinoma HEp-2 cell line surface, using lectin wheat germ agglutinin (WGA) and on mitochondrial activity, for the same cell line, using MitoTracker. The photosensitizer Aluminum Phthalocyanine Tetrasulfonate (AlPcS4) was administrated at 10 µM/mL, followed by an incubation period for its accumulation in the tumor cells, which were irradiated with laser diode λ = 685 nm and energy density of 4.5 J/cm2. Our results indicated that, after Photodynamic Therapy (PDT), it was observed N-acetyl glucosamine terminal glycoprotein expression and mitochondrial O2 production, compared to the control group. Based on these results, we suggest that PDT influences the O2 mitochondrial production and the presence of surface glycoproteins N-acetyl glucosamine terminals.

Method of isolation and characterization of Girardia tigrina stem cells.

Tissue regeneration is widely studied due to its importance for understanding the biology of stem cells, aiming at their application in medicine for therapeutic and various other purposes. The establishment of experimental models is necessary, as certain invertebrates and vertebrates have different regeneration abilities depending on their taxon position on the evolutionary scale. Planarians are an efficacious in vivo model for stem cell biology, but the correlation between planarian cellular and molecular neoblast pluripotency mechanisms and those of mammalian stem cells is unknown. The present study had the following objectives: i) Establish Girardia tigrina cell culture, ii) determine the time required for complete cell disintegration and iii) obtain neoblasts by cell subdivision. Twenty-four specimens were deprived of food for seven days. After this time, disintegration was performed by incubation protected at three temperatures for 48 h in an antibiotic, antimycotic and trypsin solution, after which the suspension was homogenized and centrifuged. Histopaque® 1077 was used for cell separation and interphases were collected and monitored by optical and fluorescence microscopy. Optical microscopy analysis informed the nucleus-to-cytoplasm ratio, cell morphology and cell size. Under fluorescence microscopy, interphase 1 (I1) was subdivided into two groups and neoblasts were marked for characterization; one group was stained with 4',6-diamidino-2-phenylindole and the other was immunolabeled with octamer-binding transcription factor 4 (OCT4) and isolated and observed after 10 days of cultivation. Neoblasts predominated in I1 with a small amount of other cell types. In conclusion, sample disintegration with a trypsin and antibiotic solution was effective at 18˚C and Iscove's modified Dulbecco's medium supplemented with fetal bovine serum was adequate for the establishment of primary cell cultures after 48-h incubation and centrifugation. Antibody anti-OCT4 was used for the characterization of stem cells and was successfully labeled with concentrated neoblasts on interphase 1.

A study of low power laser on the regenerative process of Girardia tigrina (Girard,1850) (Turbellaria; Tricladida; Dugesiidae).

The mechanism of regeneration does not start to restore the wound until its corresponding epimorphic phase. A bioestimulation of tissues and cells by laser radiation depends on the wavelength, on the dose, and on the intensity of the light. The goal of this work was to verify the effect of the low power laser at 660 nm on the regenerative process of Girardia tigrina. The specimens were maintained in the laboratory under a temperature ranging from 19 degrees up to 24 degrees C for 21 days. The planarians were anesthetized by placing them on ice and then cut them with a scalpel. The three treatments were as following: animals individually irradiated with 14 sessions with 1 minute duration (treatment 1), 14 sessions with 3 minutes duration (treatment 2), and without irradiation (control). The planarians were amputated and divided in three study treatments: a control group (without radiation), and two other treatments: irradiated for 1 minute, and irradiated for 3 minutes. The animals were irradiated with diode laser (660 nm) with 3.3 +/- 0.3 mW of power, using 0.94 mW.mm-2 power density for each irradiation procedure. During the experiment, 14 irradiation sessions were undertaken. The specimens were fixed in Bouin, and stained with hematoxyline and eosin. From observation and histological analysis, it was possible to assess the effects of interaction between laser and tissue. The head fragment after 1 minute of irradiation presented a better organized tissue scheme, when compared with the other treatments. Aspects of the body fragments submitted to 3 minutes of light treatment were very similar to fragments that had not been injured. It can be concluded that there are changes in the quality of regeneration when treated with low power laser under the conditions mentioned above.

A study of low power laser on the regenerative process of Girardia tigrina (Girard, 1850) (Turbellaria; Tricladida; Dugesiidae) / Um estudo de laser de baixa potência no processo regenerativo de Girardia tigrina (Girard, 1850) (Turbellaria; Tricladida; Dugesiidae)

The mechanism of regeneration does not start to restore the wound until its corresponding epimorphic phase. A bioestimulation of tissues and cells by laser radiation depends on the wavelength, on the dose, and on the intensity of the light. The goal of this work was to verify the effect of the low power laser at 660 nm on the regenerative process of Girardia tigrina. The specimens were maintained in the laboratory under a temperature ranging from 19° up to 24 °C for 21 days. The planarians were anesthetized by placing them on ice and then cut them with a scalpel. The three treatments were as following: animals individually irradiated with 14 sessions with 1 minute duration (treatment 1), 14 sessions with 3 minutes duration (treatment 2), and without irradiation (control). The planarians were amputated and divided in three study treatments: a control group (without radiation), and two other treatments: irradiated for 1 minute, and irradiated for 3 minutes. The animals were irradiated with diode laser (660 nm) with 3.3 ± 0.3 mW of power, using 0.94 mW.mm-2 power density for each irradiation procedure. During the experiment, 14 irradiation sessions were undertaken. The specimens were fixed in Bouin, and stained with hematoxyline and eosin. From observation and histological analysis, it was possible to assess the effects of interaction between laser and tissue. The head fragment after 1 minute of irradiation presented a better organized tissue scheme, when compared with the other treatments. Aspects of the body fragments submitted to 3 minutes of light treatment were very similar to fragments that had not been injured. It can be concluded that there are changes in the quality of regeneration when treated with low power laser under the conditions mentioned above.

O mecanismo de regeneração não restaura a ferida até sua fase epimórfica correspondente. A bioestimulação de tecidos e células por radiação laser depende do comprimento de onda, da dose e intensidade de luz. O objetivo deste trabalho foi verificar o efeito do laser de baixa potência (660 nm) no processo regenerativo de Girardia tigrina. Estes vermes têm uma elevada capacidade de regeneração. Os espécimes foram mantidos em laboratório sob uma temperatura de 19° a 24 °C por 21 dias. As planárias foram anestesiadas com gelo e amputadas com bisturi. Os 3 tratamentos foram compostos de: animais individualmente irradiados com 14 sessões de 1 minuto de duração (tratamento 1), com 14 sessões de três minutos de duração (tratamento 2) e não irradiados (controle). Elas foram irradiadas com laser diodo (660 nm) 3,3 ± 0,3 mW potência, usando 0.94 mW.mm-2 de densidade de potência para cada irradiação. Durante o experimento foram realizadas 14 sessões. Os espécimes foram fixados em Bouin e corados com hematoxilina e eosina. Com base nas observações e análise histológica, foi possível avaliar os efeitos da interação entre laser e tecido. O fragmento cabeça após 1 minuto de irradiação mostrou um tecido melhor organizado quando comparado com outros tratamentos. Os aspectos dos fragmentos corpo após 3 minutos de irradiação foram muito similares para os fragmentos que não sofreram injúria. Existem mudanças na qualidade de regeneração quando tratadas com laser nas condições acima mencionadas.

Morphometry of the eyes in regenerant of genus Dugesia (Platyhelminthes, Turbellaria, Dugesiidae)

A técnica de fotodocumentação foi utilizada para observar semanalmente os olhos de regenerantes de três espécies de Dugesia até o final do processo regenerativo. Os dados morfométricos analisados foram baseados na mensuração de 96 exemplares das espécies: Dugesia tigrina, D. schubarti e D. anderlani. Uma análise comparativa foi realizada por meio de representação gráfica de parâmetros estatísticos dos seguintes caracteres morfométricos: comprimento dos eixos transversal e longitudinal da mácula alba, cálice do pigmento e abertura ocular. D. schubarti apresenta um aspecto interessante: a média da abertura ocular na fase do processo regenerativo é maior do que nos exemplares originais.