Fatores litogênicos em pacientes com litíase urinária de Fortaleza, Ceará

Lithiasis is considered a public health issue due to its high prevalence and rates of recurrence. Objective: To identify risk factors for lithiasis in kidney stone patients from Fortaleza, Brazil. In the first stage of the study, the medical records of 197 patients with urinary lithiasis covering the period 1996 2006 were analyzed with regard to clinical and metabolic data. In the second stage, 340 kidney stones were submitted to morphological examination under 10x magnification. According to the external morphology and the cut surface, the stones were classified as pure or mixed, and major and minor components were identified. In addition, the stone fragments of 25 patients treated with lithotripsy were submitted to morphological analysis. In the third stage, a subsample of 50 stones was used in a double-blind comparison of morphological and chemical findings. Results were expressed as concordant, partly concordant (discordant for minor components) or discordant (discordant for major components). The average age of first symptoms was 35.8±13.3 years, with no significant difference between the genders. The male/female ratio was 1:1.7. Recurrence was reported in 53.3% of cases. The main metabolic changes observed were hypernatriuria (80.7%), hypercalciuria (48.7%), low urine volume (43.7%), hyperoxaluria (30.5%) and hyperuricosuria (17.3%). Pure stones represented 34.7% of the total sample of 340 stones. The most common route of elimination was spontaneous for pure stones (49.1%) and surgical for mixed stones (50.5%). Pure stones consisted most frequently of calcium oxalate (OxCa) (59.3%) and uric acid (UA) (23.7%), the former prevalent in women, the latter prevalent in men. The most frequently observed component in mixed stones was OxCa (67.1%), followed by carbapatite (11.2%) and struvite (7.9%). The main components were OxCa and UA for men, and carbapatite and struvite for women. Nearly half (48%) the 25 analyzed fragments were pure, consisting of calcium oxalate dihydrate (COD) (56%), calcium oxalate monohydrate (COM) (48%), phosphate (32%) and UA (20%). Four patients (16%) had infectious stones. In the chemical analysis of the subsample of 50 stones, the most 64 frequently observed major components were calcium (70%), oxalate (66%), ammonium (56%), urate (28%) and carbonate (24%). In the morphological analysis, the main components were calcium and magnesium phosphate (32%), COM (24%), UA (20%), COD (18%) and cystine (6%). Morphological and chemical findings were totally concordant for 38% of the stones, partly concordant in 52% and discordant in 10%. Conclusion: The risk factors for lithiasis in kidney stone patients from Fortaleza (Brazil) were hyperoxaluria, hypercalciuria with or without hypernatriuria, hyperuricosuria and low urine volume

Propriedades bioativas dos polissacarídeos sulfatados da alga comestível Gracilaria birdiae

Red marine algae of the genus Gracilaria synthesize sulfated polysaccharides (PS) bioactive. But many of these PS were not properly assessed, as is the case of PS synthesized by edible seaweed Gracilaria birdiae. Previous studies showed that sulfated galactans this alga has anti-inflammatory effect. In this work, a galactan (GB) of G. birdiae was obtained and evaluated by different tests. GB showed anticoagulant activity in APTT assay. GB showed no toxicity to normal cells (3T3), but inhibited the survival of cells of adenocarcinoma of the cervix (HeLa) and human pancreatic cancer (Panc-1) 80% (1.5 mg / ml). GB was not able to hijack the OH radical or the superoxide radical. However, showed activity electron donor in two different tests and presented iron chelator activity (70% and 1.0 mg / ml) and Copper (70% at 0.5 mg / ml). The presence of a higher GB promotes formation of crystals of calcium oxalate dihydrate small size, which is less aggressive, because GB is able to interact with and stabilize the crystal that form. Furthermore, GB (2.0 mg / mL) was not cytotoxic to human renal cells (HEK-293). The data lead us to propose that GB has a great potential for the treatment of urolithiasis

Quintosana e seus derivados conjugados com ácido gálico: avaliação de seu potencial antioxidante e de interferência na formação de cristais de oxalato de cálcio

Chitin is the second most abundant polysaccharide in nature and its derivative chitosan has been widely studied due to its unique chemical and pharmacological properties. However, studies show that when this molecule is used as food, drug, etc. it tends to accumulate in renal tissue and promotes an increase in calcium excretion. Nevertheless, the effect of chitosan on the formation of calcium oxalate (OxCa) crystals has never been evaluated. The formation of kidney stones (urolithiasis) is the disease that most often affects the kidneys and the urinary system. In addition, this is a disease with high prevalence and recurrence. Many molecules with antioxidant activity have been shown to decrease the potential for in vitro OxCa crystals formation. Thus, the aim of this study was to evaluate the effect of low molecular weight chitosan and its derivatives conjugated to gallic acid (AG) as antioxidant and inhibitor of OxCa crystals formation. The physico-chemical analysis confirmed the identity of chitosan. This molecule was subjected to five antioxidant tests and showed an excellent copper chelating activity. However, chitosan did not show other significant antioxidant activity. When chitosan was subjected to in vitro crystal formation tests, it increased the number of OxCa monohydrate crystals, modified the morphology of the crystals, modified the proportions between populations of crystals in solution and increased the zeta potential of these crystals formed. Four molecules of chitosan conjugated with GA were obtained. The physico-chemical analysis confirmed that chitosan and AG were covalently bonded. However, the amount of GA liked to chitosan did not increase even when 10 times more GA was used in experiment. When these derivatives were subjected to antioxidant tests, all chitosan conjugates showed higher antioxidant potential than their precursors. However, they showed different activity between them, which indicating that the position where AG is conjugated is an important factor for chitosan-GA activity. When conjugated chitosans were submitted to in vitro crystal formation tests, a reduction in the crystals number was observed when compared with those formed in the presence of unconjugated chitosan. Chitosan has a strong capacity for inducing OxCa monohydrate crystal formation, as well as modify their morphology and zeta potential. Over all, the process of conjugating AG to chitosan led to an increase in antioxidant potential of this molecule and was also able to decrease its capacity of inducing in vitro crystal formation

Extração, caracterização e prospecção de atividades farmacológicas de polissacarídeos sulfatados da macroalga verde Caulerpa prolifera

This study aimed to extract, characterize and conduct a prospective analysis of pharmacological activities of sulfated polysaccharides from green seaweed Caulerpa prolifera. Seven fractions (CP-0.3/CP-0.5/CP-0.7/CP-0.9/CP-1.1/CP-1.5/CP-2.0) were obtained from C. prolifera by alkaline proteolysis followed by sequential precipitation in acetone. The physicochemical analyzes indicated that C. prolifera synthesizes a homogalactan (CP-0.9) and different populations of sulfated heteropolysaccharides. In the analysis of anticoagulant activity, all fractions except CP-0.3, influenced the intrinsic coagulation pathway. All fractions showed antioxidant activity in six different assays being more pronounced in hydrogen peroxide scavenging assay, especially CP-0.3, CP-0.7 and CP-0.9 (which obtained 61% of hydrogen peroxide scavenging), in ferric chelation assay (especially CP-0.9 with 56% chelation) and cupric chelation assay (especially CP-2.0 with 78% chelation). With respect to immunomodulatory activity, the presence of CP-0.3, CP-0.7 and CP-0.9 showed an immunogenic potential, increasing the production of nitric oxide (NO) by 48, 142 and 163 times, respectively. Conversely, the NO synthesis fell 73% after the activation of macrophages by LPS, incubated concurrently with CP-2.0. The anti-adipogenic activity of the fractions was also evaluated and CP-1.5 was able to reduce the differentiation of pre-adipocytes (3T3-L1) into adipocytes by 60%, without affecting the cell viability. The fractions CP-0.3, CP-0.5 and CP-0.9 reduced the viability of the HeLa cells (human cervical adenocarcinoma) by 55% and CP-1.5 reduced the viability of the 786-0 cells (human renal adenocarcinoma) by 75%. Leishmanicidal activity and microbicide effect against Carbapenem-resistant Klebsiella pneumoniae (KPC) have not been identified. However, the viability of Staphylococcus epidermidis was reduced by 23.8% in the presence of CP -1.5. All fractions were able to change the formation of calcium oxalate crystals. CP-0.3, CP-0.5 and CP-1.1 only promoted the formation of COD type crystals with a very small size (1 μm). Confocal microscopy and zeta potential data of crystals formed in the presence of the samples showed that the polysaccharides present in the fractions must interact with calcium ions present throughout the crystal lattice, affecting the growth and morphology of crystals The results described herein indicate that the fractions rich in polysaccharides obtained from the green seaweed C. prolifera present a multi therapeutic potential, and subsequent purification steps, as well as research on the mechanisms of action by which these polymers act should be investigated.

Avaliação das atividades farmacológicas de fucoidans obtidos do extrato bruto de fucoidan comercial de Fucus vesiculosus

Fucoidan is a term used to define heteropolysaccharides that are composed of less than 90% L-fucose. The exception to this rule is the homofucoidan obtained from the seaweed Fucus vesiculosus. This fucoidan can be purchased from SIGMA Co. and have been used in various research for evaluation of their pharmacological activities. However, it is not a pure molecule. In fact, it is a mix of several fucoidan molecules. In this work, were obtained, from acetone precipitation, and biochemically characterized, four fucoidan molecules from SIGMA-ALDRICH Co. fucoidan to evaluate their anticoagulant, antioxidant, antiadipogenic, immunomodulatory and antiurolithiatic activities. In anticoagulant activity, evaluated by aPTT assay, fucoidans F0.9, F1.1 and F2.0 increased eightfold the coagulation time, compared to the control, when a mass of 10 μg was used. To PT test, only fucoidan F0.9 was capable of increase the coagulation time, compared to control. In the total antioxidant capacity assay (TAC), the fucoidan F2.0 showed 400 ascorbic acid equivalents, while fucoidan F0.5, the lest effective, 38 equivalents. In respect to the effect on pre-adipocyte cell lines (3T3-L1) adipogenesis, was observed that fucoidan F1.1 and F2.0 reduced the adipogenesis and this effect was associated to the reduction in the expression of regulatoy proteins C/EBPα, C/EBPβ and PPARγ. On the other hand, fucoidans F0.5 and F0.9 induced increased expression of these regulatory proteins. Furthermore, fucoidan F2.0 induced hydrolysis of triglycerides present in the interior of adipocytes. The immunomodulatory effect was evaluated and observed that the presence of fucoidans F0.5 , F1.1 and F2.0 significantly reduced the production of nitric oxide by activated macrophages with LPS specially fucoidan F2.0 that in 100 μg/mL, reduced about 55% the effect caused by LPS. Relative to the effect upon the formation of calcium oxalate crystals, fucoidan F0.5 was more effective in reduce the aggregation of the crystals and this effect it was not significantly different regarding the effect caused by the crude. Besides, fucoidan F0.5 only promoted the formation of COD type crystals, while fucoidans F1.1 and F2.0 did not influence the formation of crystals compared with the control. The results described in this study indicate that the commercial crude fucoidan of Fucus vesiculosus it’s a mix of several fucoidan which, in turn, have different chemical compositions besides having different pharmacological activities. The use of these fucoidans it´s indicated according the pharmacological activity to be evaluated.

Avaliação das atividades farmacológicas de fucoidans obtidos do extrato bruto de fucoidan comercial de Fucus vesiculosus

Fucoidan is a term used to define heteropolysaccharides that are composed of less than 90% L-fucose. The exception to this rule is the homofucoidan obtained from the seaweed Fucus vesiculosus. This fucoidan can be purchased from SIGMA Co. and have been used in various research for evaluation of their pharmacological activities. However, it is not a pure molecule. In fact, it is a mix of several fucoidan molecules. In this work, were obtained, from acetone precipitation, and biochemically characterized, four fucoidan molecules from SIGMA-ALDRICH Co. fucoidan to evaluate their anticoagulant, antioxidant, antiadipogenic, immunomodulatory and antiurolithiatic activities. In anticoagulant activity, evaluated by aPTT assay, fucoidans F0.9, F1.1 and F2.0 increased eightfold the coagulation time, compared to the control, when a mass of 10 μg was used. To PT test, only fucoidan F0.9 was capable of increase the coagulation time, compared to control. In the total antioxidant capacity assay (TAC), the fucoidan F2.0 showed 400 ascorbic acid equivalents, while fucoidan F0.5, the lest effective, 38 equivalents. In respect to the effect on pre-adipocyte cell lines (3T3-L1) adipogenesis, was observed that fucoidan F1.1 and F2.0 reduced the adipogenesis and this effect was associated to the reduction in the expression of regulatoy proteins C/EBPα, C/EBPβ and PPARγ. On the other hand, fucoidans F0.5 and F0.9 induced increased expression of these regulatory proteins. Furthermore, fucoidan F2.0 induced hydrolysis of triglycerides present in the interior of adipocytes. The immunomodulatory effect was evaluated and observed that the presence of fucoidans F0.5 , F1.1 and F2.0 significantly reduced the production of nitric oxide by activated macrophages with LPS specially fucoidan F2.0 that in 100 μg/mL, reduced about 55% the effect caused by LPS. Relative to the effect upon the formation of calcium oxalate crystals, fucoidan F0.5 was more effective in reduce the aggregation of the crystals and this effect it was not significantly different regarding the effect caused by the crude. Besides, fucoidan F0.5 only promoted the formation of COD type crystals, while fucoidans F1.1 and F2.0 did not influence the formation of crystals compared with the control. The results described in this study indicate that the commercial crude fucoidan of Fucus vesiculosus it’s a mix of several fucoidan which, in turn, have different chemical compositions besides having different pharmacological activities. The use of these fucoidans it´s indicated according the pharmacological activity to be evaluated.