Chlorophyll treatment combined with photostimulation increases glycolysis and decreases oxidative stress in the liver of type 1 diabetic rats

Photodynamic therapy (PDT) promotes cell death, and it has been successfully employed as a treatment resource for neuropathic complications of diabetes mellitus (T1DM) and hepatocellular carcinoma. The liver is the major organ involved in the regulation of energy homeostasis, and in pathological conditions such as T1DM, changes in liver metabolic pathways result in hyperglycemia, which is associated with multiple organic dysfunctions. In this context, it has been suggested that chlorophyll-a and its derivatives have anti-diabetic actions, such as reducing hyperglycemia, hyperinsulinemia, and hypertriglyceridemia, but these effects have not yet been proven. Thus, the biological action of PDT with chlorophyll-a on hepatic parameters related to energy metabolism and oxidative stress in T1DM Wistar rats was investigated. Evaluation of the acute effects of this pigment was performed by incubation of isolated hepatocytes with chlorophyll-a and the chronic effects were evaluated by oral treatment with chlorophyll-based extract, with post-analysis of the intact liver by in situ perfusion. In both experimental protocols, chlorophyll-a decreased hepatic glucose release and glycogenolysis rate and stimulated the glycolytic pathway in DM/PDT. In addition, there was a reduction in hepatic oxidative stress, noticeable by decreased lipoperoxidation, reactive oxygen species, and carbonylated proteins in livers of chlorophyll-treated T1DM rats. These are indicators of the potential capacity of chlorophyll-a in improving the status of the diabetic liver.

Aquaculture biological waste as culture medium to cultivation of Ankistrodesmus gracilis (Reinsch) Korshikov / Resíduos biológicos de aquicultura como meio de cultura para o cultivo de Ankistrodesmus gracilis (Reinsch) Korshikov

Abstract Current study investigated the effectiveness of different macrophytes as culture media for Ankistrodesmus gracilis in laboratory conditions. Significant difference (p < 0.05) was reported in cell density with regard to conventional culture medium (CHU12) and macrophytes culture media. Mean cell density in NPK, Eichhornia crassipes and E. azurea media was higher (p < 0.05) than in conventional culture medium. Chlorophyll-a was higher than 1 g.L-1, except in CHU12 (0.7 ± 0.4 g.L-1) and T. domingensis (0.8 ± 0.3 g.L-1) media. Nitrate decreased sharply as from the 7th-day of the experiment. Ammonium and total phosphorus were highest in culture media and ranged between 0.4 g.L-1 (P. cordata medium) and 1.7 g.L-1 (CHU12 medium) for ammonium, and between 0.8 g.L-1 (CHU12 medium) and 1.9 g.L-1 (T. domingensis medium) for total phosphorus. Results revealed inorganic fertilizer and macrophytes combined with vitamins may be effective as culture media and strongly supports the growth of Ankistrodesmus gracilis, since cell density and biochemical composition are similar to or higher than conventional culture medium (CHU12). Macrophyte is a tool for aquaculture since biological wastes may be used with nutrients to improve the cultivation of microalgae.

Resumo O presente estudo investigou a eficácia na utilização de diferentes macrófitas como meio de cultura para Ankistrodesmus gracilis em condições laboratoriais. Foi observada diferença significativa (p < 0,05) entre a densidade celular em relação aos meios de cultura convencional (CHU12) e de macrófitas. Os meios de cultura com Eichhornia crassipes, E. azurea e NPK apresentaram densidade celular maiores (p < 0,05) que o meio de cultura convencional. Os teores de clorofila-a foram superiores a 1 g.L-1, exceto nos meios de cultura CHU12 (0.7 ± 0.4 g.L-1) e de T. domingensis (0.7 ± 0.3 g.L-1). O nitrato diminuiu acentuadamente a partir do 7º dia de experimento. Dentre os nutrientes, amônia e o fósforo total foram os mais elevados nos meios de cultura, variando entre 0.4 g.L-1 (meio de P. cordata) a 1,7 g.L-1 (meio CHU12) para amônia e, entre 0.8 g.L-1 (meio CHU12) e 1,9 gL-1 (meio de T. domingensis) para o fósforo total. Os resultados do presente estudo mostraram que o uso de fertilizantes inorgânicos e macrófitas, combinados com vitaminas, pode ser eficaz como meio de cultura no crescimento de Ankistrodesmus gracilis, uma vez que a densidade celular e a composição bioquímica foram semelhantes ou superiores ao meio de cultura convencional (CHU12). As macrófitas são ferramentas a serem adotadas na aquicultura, desde que os resíduos biológicos podem ser usados como nutrientes para melhorar o cultivo de microalgas.

Efeito do déficit hídrico em características químicas e bioquímicas da soja e na degradação da clorofila, com ênfase na formação de metabólitos incolores / Influence of drought stress on chemical and biochemical characteristics of soybean and on chlorophyll degradation, with focus on the formation of non-coloured chlorophyll catabolites

O Brasil é o segundo maior produtor e exportador de soja (Glycine max L. Merrill) no mundo. A produção é altamente dependente de fatores climáticos, incluindo a temperatura e quantidade de chuva. A soja cultivada no sul do país em 2005 sofreu déficit hídrico causado por temperaturas altas acompanhado por umidade baixa durante o estádio reprodutivo. Pouco se sabe sobre a influência do déficit hídrico na degradação da clorofila e na qualidade dos grãos em geral. Nesse trabalho foram analisadas, numa primeira etapa, as características químicas e bioquímicas de cinco amostras de soja, provenientes de três cultivares que cresceram sob déficit hídrico e que não atendiam aos padrões para comercialização por conter altas quantidades de sementes verdes. Os grãos foram analisados quanto a diversos parâmetros físico-químicos e bioquímicos incluindo a análise dos pigmentos verdes imediatamente após a colheita e após 20 meses de armazenamento. A acidez foi medida adicionalmente após 30 meses de armazenamento. A atividade de água e umidade foram 0,6-0,7 e 8,7 %-11,9 %, respectivamente, e não mudaram durante a estocagem, mas houve um aumento em acidez o que indica atividade de lipases. A atividade da lipoxigenase 1 foi significativamente prejudicada. Imediatamente após a colheita os pigmentos verdes correspondiam a feofitina a, feofitina b e pequenas quantidades de clorofila a e b, e traços de outros derivados da clorofila, em ordem decrescente. Após 20 meses de estocagem quase todos os pigmentos haviam desaparecido. O déficit hídrico provavelmente aumentou a permeabilidade das membranas, o que levou a um aumento do pH e promoveu a transformação das clorofilas para feofitinas. Na segunda etapa do trabalho foi estudado o estágio avançado da degradação natural da clorofila na qual desaparece a coloração esverdeada dos grãos. Esta etapa corresponde à formação de catabólitos incolores (NCC), mas existem controvérsias se essas substâncias são os produtos finais da degradação...

Brazil is the second largest soybean (Glycine max L. Merrill) producer and exporter in the world. The production depends on climatic factors, like temperature and rain volume. Soybeans cultivated in the south of the country in 2005 suffered drought stress imposed by adverse high ambient temperature, accompanied by low humidity during the reproductive stage. Little information is available regarding drought stress on quality of grains. In this study, firstly, chemical and biochemical characteristics of five soybean samples belonging to three cultivars grown under drought stress and did not meet standards for marketing due to high amounts of green seeds, were evaluated. Grains were analyzed for several physicochemical ad biochemical parameters, including analysis of pigment contents, immediately after harvest and after 20 months of storage at room temperature. Acidity was measured additionally after 30 month of storage. Water activity and humidity were 0.6 - 0.7 and 8.7 % - 11.9 %, respectively, and did not change during storage time, but there was an increase in acidity, which alludes to lipase activity. The activity of lipoxygenase 1 was significantly prejudiced. Immediately after harvest, green pigments corresponded mainly to pheophytin a, followed by pheophytin b, small quantities of chlorophyll b and chlorophyll a and traces of other chlorophyll derivatives. After 20 months of storage almost all green pigments had disappeared. Drought stress probably enhanced membrane permeability, which led to a lower pH and promoted transformation of chlorophylls to pheophytins. In the second part of the study, the advanced stage of natural chlorophyll degradation was investigated, in which the green colour of the seeds disappears. This stage corresponds to the formation of non-coloured chlorophyll catabolites (NCC), but there are controversies if those are the final products. Thus, the formation and degradation of NCC during soybean (Glycine max L. Merrill)...

Chlorophyll derivatives: a new photosensitizer for photodynamic therapy of cancer in mice

The in vivo photosensitizing efficacy of chlorophyll derivatives(CpD), which had been developed as a new photosensitizer, was compared with that of hematoporphyrin derivatives (HpD). A murine tumor model implanted subcutaneously with S-180 cells on the abdomen was used. The CpD or HpD was administered by intratumoral injection, and light of appropriate wavelength was irradiated on the tumor areas for 10 minutes at 1h and 24h or 24h and 48h after the injection of photosensitizer. When CpD was injected, the early irradiation group (1h and 24h) showed a 100% tumor cure rate; however, the late irradiation group (24h and 48h) showed a 60% tumor cure rate (p less than 0.01). This showed that the early irradiation with light after injection of CpD was an important factor for obtaining better results. With HpD, there was no difference in tumor cure rate between early (1h and 24h, 80%) and late irradiation (24h and 48h, 80%) groups. Thus, in early irradiation groups, the tumor cure rate using CpD (100%) was superior to that of HpD (80%) (p less than 0.05). However, in late irradiation groups, the tumor cure rate using CpD (60%) was inferior to that of HpD (80%), but this difference was not statistically significant (p greater than 0.1). Pathologic sections of these tumors were made before treatment and 48h and 3 weeks after treatment. These showed geographic necrosis at 48h after treatment and no viable tumor tissue at 3 weeks after treatment. Our results showed that CpD was as effective as HpD as a photosensitizer for in vivo photodynamic therapy.

Chlorophyll derivatives: a new photosensitizer for photodynamic therapy of cancer in mice

The in vivo photosensitizing efficacy of chlorophyll derivatives(CpD), which had been developed as a new photosensitizer, was compared with that of hematoporphyrin derivatives (HpD). A murine tumor model implanted subcutaneously with S-180 cells on the abdomen was used. The CpD or HpD was administered by intratumoral injection, and light of appropriate wavelength was irradiated on the tumor areas for 10 minutes at 1h and 24h or 24h and 48h after the injection of photosensitizer. When CpD was injected, the early irradiation group (1h and 24h) showed a 100% tumor cure rate; however, the late irradiation group (24h and 48h) showed a 60% tumor cure rate (p less than 0.01). This showed that the early irradiation with light after injection of CpD was an important factor for obtaining better results. With HpD, there was no difference in tumor cure rate between early (1h and 24h, 80%) and late irradiation (24h and 48h, 80%) groups. Thus, in early irradiation groups, the tumor cure rate using CpD (100%) was superior to that of HpD (80%) (p less than 0.05). However, in late irradiation groups, the tumor cure rate using CpD (60%) was inferior to that of HpD (80%), but this difference was not statistically significant (p greater than 0.1). Pathologic sections of these tumors were made before treatment and 48h and 3 weeks after treatment. These showed geographic necrosis at 48h after treatment and no viable tumor tissue at 3 weeks after treatment. Our results showed that CpD was as effective as HpD as a photosensitizer for in vivo photodynamic therapy.