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.

Anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide in mice

Several isatin derivatives have shown important biological activities, which have attracted interest from researchers. For this reason, the present study aimed to evaluate the anti-inflammatory and antinociceptive effects of the isatin derivative (Z)-2-(5-chloro-2-oxoindolin-3-ylidene)-N-phenyl-hydrazinecarbothioamide (COPHCT) in mice. Three doses of this compound were tested: 1.0, 2.5, and 5.0 mg/kg. The anti-inflammatory activity was assessed using the carrageenan-induced paw edema model and the zymosan-induced air pouch model. The evaluation of the antinociceptive effect was performed through the formalin test and the acetic acid-induced abdominal writhing test. The paw edema assay demonstrated that all doses of the compound showed a significant reduction of the edema in the second hour evaluated, but a better response was observed in the fourth hour. The zymosan-induced air pouch model indicated that the compound, in all doses, significantly reduced leukocyte migration and total protein concentration levels. In the formalin test, the doses 1.0, 2.5, and 5.0 mg/kg of COPHCT showed activity only in the second phase, with reduction in paw pain time of 73.61, 79.46, and 73.85%, respectively. The number of abdominal writhings decreased with the increasing dose, but only 5.0 mg/kg COPHCT exhibited a significant response, with a reduction of 24.88%. These results demonstrated the ability of this compound to interfere in the anti-inflammatory activity of edema, vascular permeability, and cell migration. In addition, its possible antinociceptive effect may be related to the dose used.

Pros and cons of insulin administration on liver glucose metabolism in strength-trained healthy mice

Non-diabetic individuals use hormones like insulin to improve muscle strength and performance. However, as insulin also leads the liver and the adipose tissue to an anabolic state, the purpose of this study was to investigate the effects of insulin on liver metabolism in trained non-diabetic Swiss mice. The mice were divided into four groups: sedentary treated with saline (SS) or insulin (SI) and trained treated with saline (TS) or insulin (TI). Training was made in a vertical stair, at 90% of the maximum load, three times per week. Insulin (0.3 U/kg body weight) or saline were given intraperitoneally five times per week. After eight weeks, tissue and blood were collected and in situ liver perfusion with glycerol+lactate or alanine+glutamine (4 mM each) was carried out. The trained animals increased their muscle strength (+100%) and decreased body weight gain (-11%), subcutaneous fat (-42%), mesenteric fat (-45%), and peritoneal adipocyte size (-33%) compared with the sedentary groups. Insulin prevented the adipose effects of training (TI). The gastrocnemius muscle had greater density of muscle fibers (+60%) and less connective tissue in the trained groups. Liver glycogen was increased by insulin (SI +40% and TI +117%), as well as liver basal glucose release (TI +40%). Lactate and pyruvate release were reduced to a half by training. The greater gluconeogenesis from alanine+glutamine induced by training (TS +50%) was reversed by insulin (TI). Insulin administration had no additional effect on muscle strength and reversed some of the lipolytic and gluconeogenic effects of the resistance training. Therefore, insulin administration does not complement training in improving liver glucose metabolism.

Effects of nanoparticles of hydroxy-aluminum phthalocyanine on markers of liver injury and glucose metabolism in diabetic mice

Photodynamic therapy, by reducing pain and inflammation and promoting the proliferation of healthy cells, can be used to treat recurrent lesions, such as diabetic foot ulcers. Studies using the photosensitizer phthalocyanine, together with the nanostructured copolymeric matrix of Pluronic® and Carbopol® for the treatment of diabetic foot ulcers and leishmaniosis lesions, are showing promising outcomes. Despite their topical or subcutaneous administration, these molecules are absorbed and their systemic effects are unknown. Therefore, we investigated the effect of the subcutaneous administration of the hydroxy-aluminum phthalocyanine hydrogel without illumination on systemic parameters, markers of liver injury, and liver energy metabolism in type 1 diabetic Swiss mice. Both the hydrogel and the different doses of phthalocyanine changed the levels of injury markers and the liver glucose release, sometimes aggravating the alterations caused by the diabetic condition itself. However, the dose of 2.23 µg/mL caused less marked plasmatic and metabolic changes and did not change glucose tolerance or insulin sensitivity of the diabetic mice. These results are indicative that the use of hydroxy-aluminum phthalocyanine hydrogel for the treatment of cutaneous ulcers in diabetic patients is systemically safe.

Sentinel node biopsy using blue dye and technetium99 in advanced gastric cancer: anatomical drainage and clinical application

Lymph node metastases are an independent prognosis factor in gastric carcinoma (GC) patients. Radical lymphadenectomy can improve survival but it can also increase surgical morbidity. As a principle, sentinel node (SN) navigation surgery can avoid unnecessary lymphadenectomy without compromising prognosis. In this pilot study, 24 patients with untreated GC were initially screened for SN navigation surgery, of which 12 were eligible. Five patients had T2 tumors, 5 had T3 tumors and 2 had T1 tumors. In 33% of cases, tumor diameter was greater than 5.0 cm. Three hundred and eighty-seven lymph nodes were excised with a median of 32.3 per patient. The SN navigation surgery was feasible in all patients, with a median of 4.5 SNs per patient. The detection success rate was 100%. All the SNs were located in N1 and N2 nodal level. In 70.9% of cases, the SNs were located at lymphatic chains 6 and 7. The SN sensitivity for nodal staging was 91.6%, with 8.3% of false negative. In 4 patients who were initially staged as N0, the SNs were submitted to multisection analyses and immunohistochemistry, confirming the N0 stage, without micrometastases. In one case initially staged as negative for nodal metastases based on SN analyses, metastases in lymph nodes other than SN were found, resulting in a 20% skip metastases incidence. This surgery is a reproducible procedure with 100% detection rate of SN. Tumor size, GC location and obesity were factors that imposed some limitations regarding SN identification. Results from nodal multisection histology and immunohistochemistry analysis did not change initial nodal staging.

Evaluation of multiplex Polymerase chain reaction utilising multiple targets in Mycobacterium tuberculosis direct test negative but culture positive cases: A potential method for enhancing the diagnosis of tuberculosis.

Purpose: To evaluate multiplex Polymerase Chain Reaction (MPCR) utilising multiple targets (IS6110, Protein b [Pab] and MPB64 genes) in Mycobacterium tuberculosis Direct Test (MTD) negative but culture positive cases and comparison of MPCR with Real-Time polymerase chain reaction (RT-PCR) for diagnosis of tuberculosis. Materials and Methods: MPCR was carried out on 28 culture positive sputum samples. Out of 28 culture positive samples, 17 were originally reported, as MTD test negative and 11 were MTD test positive, respectively. The results of MPCR were compared with RT-PCR. To check the specifi city of the tests, MPCR and RT-PCR were also evaluated with 16 non-tuberculous mycobacterial (NTM) isolates. Results: Out of 28 culture positive sputum samples, MPCR was positive in all 28/28 samples, whereas RT-PCR was positive in 27/28 samples and MTD test was originally tested positive in six sputum samples and on repeating MTD testing, fi ve more sputum samples were positive and thus total number of MTD positive were 11/28 sputum samples, respectively. All the tests were negative on evaluation with all the 16 NTMs, thus giving specifi city of 100% to all the tests; sensitivity of MPCR, RT-PCR and MTD tests were 100%, 96.42% and 39.28%, respectively, in these specifi cally selected samples. Conclusions: MPCR may be an important tool in the rapid diagnosis of tuberculosis especially in disease endemic, resource limited countries.

Identificação botânica e química de espécies vegetais de uso popular no Rio Grande do Norte, Brasil / Botanical and chemical identification of plant species of popular use in Rio Grande do Norte, Brazil

Plantas medicinais são utilizadas mundialmente como uma das principais formas de cuidado primário de saúde. No entanto, a literatura indica que muitas espécies podem apresentar composição química variável, toxicidade ou difícil identificação. O objetivo do presente estudo foi obter critérios úteis para o controle de qualidade farmacognóstico das principais espécies vegetais de uso popular no estado do Rio Grande do Norte, utilizando metodologias de análise botânica (morfodiagnose macro e microscópica) e química (triagem fitoquímica e cromatografia em camada delgada), evitando assim adulterações ou uso inadequado dessas plantas medicinais no estado. No total, sete espécies foram analisadas Acmella oleracea, Chenopodium ambrosioides, Lippia alba, Mentha piperita, Ocimum gratissimum, Peumus boldus e Rosmarinus officinalis. Diversos marcadores botânicos e fitoquímicos foram identificados, contribuindo dessa forma para a correta identificação destas espécies de plantas medicinais importantes no estado do Rio Grande do Norte.

Medicinal plants are worldwide used as one of the main forms of primary healthcare. However, the literature indicates that many species may have variable chemical composition, toxicity, or even difficult identification. The aim of this study was to obtain useful criteria for pharmacognostic quality control of the main plant species of popular use in the state of Rio Grande do Norte, using methods of botanical (macro and microscopic morphodiagnosis) and chemical (phytochemical screening and thin-layer chromatography) analysis, thus preventing adulteration or inappropriate use of these medicinal plants in the state. In total, seven species were analyzed Acmella oleracea, Chenopodium ambrosioides, Lippia alba, Mentha piperita, Ocimum gratissimum, Peumus boldus and Rosmarinus officinalis. Several botanical and phytochemical markers were identified, thereby contributing to the correct identification of these important medicinal plant species in the state of Rio Grande do Norte.

Validation of quantitative analysis method for triamcinolone in ternary complexes by UV-Vis spectrophotometry / Validação de método de análise quantitativa para a triancinolona a partir de complexo ternário por espectrofotometria de UV-Vis

Triamcinolone (TRI), a drug widely used in the treatment of ocular inflammatory diseases, is practically insoluble in water, which limits its use in eye drops. Cyclodextrins (CDs) have been used to increase the solubility or dissolution rate of drugs. The purpose of the present study was to validate a UV-Vis spectrophotometric method for quantitative analysis of TRI in inclusion complexes with beta-cyclodextrin (B-CD) associated with triethanolamine (TEA) (ternary complex). The proposed analytical method was validated with respect to the parameters established by the Brazilian regulatory National Agency of Sanitary Monitoring ANVISA). The analytical measurements of absorbance were made at 242nm, at room temperature, in a 1-cm path-length cuvette. The precision and accuracy studies were performed at five concentration levels (4, 8, 12, 18 and 20µg.mL-1. The B-CD associated with TEA did not provoke any alteration in the photochemical behavior of TRI. The results for the measured analytical parameters showed the success of the method. The standard curve was linear (r2 > 0.999) in the concentration range from 2 to 24 µg.mL-1. The method achieved good precision levels in the inter-day (relative standard deviation-RSD <3.4%) and reproducibility (RSD <3.8%) tests. The accuracy was about 80% and the pH changes introduced in the robustness study did not reveal any relevant interference at any of the studied concentrations. The experimental results demonstrate a simple, rapid and affordable UV-Vis spectrophotometric method that could be applied to the quantitation of TRI in this ternary complex.

A triancinolona (TRI) é um fármaco amplamente utilizado no tratamento de doenças inflamatórias do globo ocular e é praticamente insolúvel em água, o que limita sua utilização na forma de colírio. As ciclodextrinas (CDs) têm sido utilizadas com sucesso para aumentar a solubilidade ou velocidade de dissolução de fármacos. O presente estudo teve como objetivo a validação de uma metodologia analítica para a TRI a partir de complexos de inclusão com beta-ciclodextrina (B-CD) associada com a trietanolamina (TEA) (complexo ternário) por espectrofotometria de UV-Vis. A validação do método proposto foi realizada de acordo com os parâmetros analíticos estabelecidos pela Agência Nacional de Vigilância Sanitária (ANVISA). As análises quantitativas foram realizadas a 242nm a temperatura ambiente, utilizando cubeta de quartzo de 1cm. Os estudos de precisão e exatidão foram realizados para cinco níveis de concentração (4, 8, 12, 18 e 20µg.mL-1). A B-CD associada a TEA não alterou o comportamento fotoquímico da TRI. Os resultados da avaliação dos parâmetros analíticos demonstraram o sucesso da metodologia. A curva padrão apresentou linearidade (r2 > 0.999) na faixa de concentração de 2 a 24µg.mL-1. A metodologia apresentou bons níveis de precisão para o estudo inter dia (desvio padrão relativo-DPR <3.4%) e reprodutibilidade (DPR<3.8%). A exatidão ficou em torno de 80% e a variação de pH inserida no estudo de robustez não revelou uma interferência significativa em todas as concentrações estudadas. Os resultados experimentais demonstraram um simples, rápido e viável método de espectrofotometria de UV-Vis com aplicabilidade para a análise quantitativa da TRI a partir do complexo ternário.

Baroreflex function in conscious rats submitted to iron overload

Our hypothesis is that iron accumulated in tissue, rather than in serum, may compromise cardiovascular control. Male Fischer 344 rats weighing 180 to 220 g were divided into 2 groups. In the serum iron overload group (SIO, N = 12), 20 mg elemental iron was injected ip daily for 7 days. In the tissue iron overload group (TIO, N = 19), a smaller amount of elemental iron was injected (10 mg, daily) for 5 days followed by a resting period of 7 days. Reflex heart rate responses were elicited by iv injections of either phenylephrine (0.5 to 5.0 µg/kg) or sodium nitroprusside (1.0 to 10.0 µg/kg). Baroreflex curves were determined and fitted to sigmoidal equations and the baroreflex gain coefficient was evaluated. To evaluate the role of other than a direct effect of iron on tissue, acute treatment with the iron chelator deferoxamine (20 mg/kg, iv) was performed on the TIO group and the baroreflex was re-evaluated. At the end of the experiments, evaluation of iron levels in serum confirmed a pronounced overload for the SIO group (30-fold), in contrast to the TIO group (2-fold). Tissue levels of iron, however, were higher in the TIO group. The SIO protocol did not produce significant alterations in the baroreflex curve response, while the TIO protocol produced a nearly 2-fold increase in baroreflex gain (-4.34 ± 0.74 and -7.93 ± 1.08 bpm/mmHg, respectively). The TIO protocol animals treated with deferoxamine returned to sham levels of baroreflex gain (-3.7 ± 0.3 sham vs -3.6 ± 0.2 bpm/mmHg) 30 min after the injection. Our results indicate an effect of tissue iron overload on the enhancement of baroreflex sensitivity.

Cardiovascular responses to locomotor activity and feeding in unrestrained three-toed sloths, Bradypus variegatus

Heart rate (HR) and systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure were recorded by biotelemetry in nine conscious unrestrained sloths for 1 min every 15 min over a 24-h period. The animals were allowed to freely move in an acoustically isolated and temperature-controlled (24 ± 1ºC) experimental room with light-dark cycle (12/12 h). Behavior was closely monitored through a unidirectional visor and classified as resting (sitting or suspended), feeding (chewing and swallowing embauba leaves, Cecropia adenops), or locomotor activity around the tree trunk or on the room floor. Locomotor activity caused statistically significant increases in SBP (+8 percent, from 121 ± 22 to 131 ± 18 mmHg), DBP (+7 percent, from 86 ± 17 to 92 ± 10 mmHg), MBP (+8 percent, from 97 ± 19 to 105 ± 12 mmHg), and HR (+14 percent, from 84 ± 15 to 96 ± 15 bpm) compared to resting values, indicating a possible major influence of the autonomic nervous system on the modulation of cardiac function during this behavior. During feeding, the increase in blood pressure was even higher (SBP +27 percent, from 119 ± 21 to 151 ± 21 mmHg; DBP +21 percent, from 85 ± 16 to 103 ± 15 mmHg; MBP +24 percent, from 96 ± 17 to 119 ± 17 mmHg), while HR remained at 14 percent (from 84 ± 15 to 96 ± 10 bpm) above resting values. The proportionally greater increase in blood pressure than in HR during feeding suggests an increase in peripheral vascular resistance as part of the overall response to this behavior.

The effect of feeding on the respiratory activity of the sloth

The aim of the present study was to confirm whether feeding influences the resting breathing rate and to observe possible alterations in blood gas and pH levels produced by feeding in unanesthetized sloths (Bradypus variegatus). Five adult male sloths (4.1 ± 0.6 kg) were placed daily in an experimental chair for a period of at least 4 h for sitting adaptation. Five measurements were made for each sloth. However, the sloths one, two and five were studied once and the sloths three and four were studied twice. Breathing rate was determined with an impedance meter and the output signal was digitized. Arterial blood samples were collected for blood gas analysis with a BGE electrolytes analyzer and adjusted for the animal's body temperature and hemoglobin content. The data are reported as mean ± SD and were collected during the resting period (8:00-10:00 h) and during the feeding period (16:00-18:00 h). The mean breathing rate increased during mastication of ymbahuba leaves (rest: 5.0 ± 1, feeding: 10 ± 1 bpm). No significant alterations were observed in arterial pH (rest: 7.42 ± 0.05, feeding: 7.45 ± 0.03), PCO2 (rest: 35.2 ± 5.3, feeding: 33.3 ± 4.4 mmHg) or PO2 (rest: 77.5 ± 8.2, feeding: 78.4 ± 5.2 mmHg) levels. These results indicate that in unanesthetized sloths 1) feeding evokes an increase in breathing rate without a significant change in arterial pH, PCO2 or PO2 levels, and 2) the increase in breathing rate produced by feeding probably is due to the act of mastication

Neuronal protein biosynthesis by neonatally malnourished and nutritionally recovered rats

1. The objective of the present study was to examine the reversibility of the effect of neonatal malnutritiona on in vivo neuronal protein synthesis. 2. Neonatal malnutrition was achieved by letting the pups suckle mothers fed a low casein diet (6.4%) which decreased the amount of milk produced. The control group was composed of pups from mothers fed a 17% casein diet and kept on the same diet after weaning. A group of pups was undernourished during the suckling period, as above, but fed a 17% casein dietafter weaning (recovered group). All animals were killed at 60 days of age. 3. Neuronal cell bodies were separated from forebraim by mechanical disruption of the tissue and centrifugation in a sucrose gradient. 4. Protein biosynthesis was measured by the incorporation of [14C] - leucine into proteins of neuronal cell bodies, after correcting for the specific activity of the precursor. 5. Forebrain cell number and cell size were evaluated by DNA content and the protein/ DNA ratio, respectively. 6. The forebrains of the recovered group weighed less than those of the controls, perhaps due to a lower number of cells, which exhibited an average size similar to that of the control group. 7. The rate of protein biosynthesis was similar in malnourished and recovered groups, but both were only 57% of the rate of the control group. 8. We conclude that neonatal malnutrition promotes an irreversible adverse effect on in vivo neuronal protein synthesis by rats