Histopathological and immunohistochemical study of antiosteoporotic efficacy of the earthworm Allolobophora caliginosa extract in orchiectomized rats / Estudio histopatológico e inmunohistoquímico de la eficacia antiosteoporótica del extracto de lombriz de tierra Allolobophora caliginosa en ratas orquiectomizadas

SUMMARY: Osteoporosis is a bone condition marked by a loss of bone mass and a disruption of bone microarchitecture. Men lose bone density as they age, resulting in brittle bones. The loss of free testosterone is one of the key factors. The objective of present study was to evaluate Allolobophora caliginosa extract (AcE) for its anti-osteoporotic and antiapoptotic activity in orchiotomized rat model at two different dose levels. Twenty eight male rats were divided into two groups. The first group represented sham operated rats while the second group underwent bilateral orchidectomy (OCX). After one week of recovery from orchidectomy surgery, the second group was randomly subdivided into 3 subgroups. The first OCX subgroup was administered orally distilled water daily for 10 weeks. The other two OCX subgroups were administered AcE (100 or200 mg/kg body weight/day) orally for 10 weeks. Orchiectomy induces remarkable loss of the cortical as well as trabecular bone loss; which, could be counterbalanced by Allolobophora caliginosa extract (AcE) that prevented cortical as well as trabecular bone loss. Allolobophora caliginosa extract (AcE) at Dose 200 mg/kg/day was found to be effective at a highly significant level in osteoporotic bone, as determined by histological images and immunohistochemical study, where Dose (100 mg/kg/day) was found to be moderately significant.In the present study, it is suggested that AcE may inhibit steroid-induced osteoblasts apoptosis, potentially via upregulation of Bcl-2 and downregulation of caspase-3. Allolobophora caliginosa extract demonstrates anti-apoptotic and anti-oxidant properties. Therefore, AcE may be used for the prevention of steroid-induced bone damage.

RESUMEN: La osteoporosis es una afección ósea caracterizada por una pérdida de masa ósea y una alteración de la microarquitectura ósea. Los hombres pierden densidad ósea a medida que envejecen, lo que resulta en huesos quebradizos. La pérdida de testosterona libre es factor clave en este proceso. El objetivo del presente estudio fue evaluar el extracto de Allolobophora caliginosa (AcE) debido a su actividad antiosteoporótica y antiapoptótica en un modelo de rata orquiectomizadas con dos niveles de dosis diferentes. Se dividieron veintiocho ratas macho en dos grupos. El primer grupo incluyó ratas con operación simulada, mientras que el segundo grupo se sometió a orquidectomía bilateral (OCX). Después de una semana de recuperación de la orquidectomía, el segundo grupo fue subdividido en 3 subgrupos. Al primer subgrupo de OCX se administró diariamente agua destilada por vía oral durante 10 semanas. Los otros dos subgrupos de OCX se administraron por vía oral AcE (100 o 200 mg / kg de peso corporal / día) durante 10 semanas. La orquidectomía induce una pérdida notable del hueso cortical y trabecular; el cual podría ser contrarrestado por el extracto de Allolobophora caliginosa (AcE) que previno la pérdida de hueso tanto cortical como trabecular visualizado en imágenes histológicas y estudio inmuno- histoquímico, donde se encontró que la dosis (100 mg / kg / día) era moderadamente significativa. En el presente estudio, se sugiere que la AcE puede inhibir la apoptosis de los osteoblastos inducida por esteroides, potencialmente a través de la regulación al alza de Bcl 2 y la regulación a la baja de caspasa 3. El extracto de Allolobophora caliginosa demuestra propiedades anti apoptóticas y antioxidantes. Por lo tanto, AcE puede usarse para la prevención del daño óseo inducido por esteroides.

Effect of powdered shells treatment of the snail Megalobulimus lopesi on wounds of diabetic rats

Abstract Purpose: To analyzed the healing effect of the powdered shell of the Megalobulimus lopesi snail on wounds of diabetic rats, since in non-diabetic rats the powdered shell presented healing potential. Methods: Seventy-two Wistar rats (Rattus norvegicus albinus) were divided into three groups: Control group (GC.diab), no therapeutic intervention on the wound; Vehicle's Control group, topical via, in diabetic rats (GCvt.diab): Powder Shell Group (PC) applied topically (GPCvt.diab): Experimental group was administered topically shortly after wound dressing and once a day during the experimental period (3, 7, 14 and 21 days) the composition containing the powdered shell of the snail. The following variables related to the healing potential were analyzed: macroscopic one, where the capacity of reduction of the wound area was evaluated; histological analysis in HE, angiogenic activity, morphometric analysis (re-epithelization), leukocyte inflammatory infiltrate; leukocyte count and also differentiation in peripheral blood. Results: The topical application in wounds of diabetic rats presented healing activity, accelerating wound closure, stimulating angiogenesis and being pro-inflammatory in the early and anti-inflammatory stages in the final times of the healing process. Conclusion: The topical administration of the powdered shell on wounds of diabetic patients becomes a therapeutic option of low cost, with ease in the administration and access as well.

Beetle (Ulomoides dermestoides) fat improves diabetes: effect on liver and pancreatic architecture and on PPARgamma expression

Ulomoides dermestoides is a beetle traditionally consumed to treat diabetes. In this study, we performed a composition analysis of U. dermestoides to obtain the principal fractions, which were used to assess the effect on glycemia, liver and pancreatic architecture, and PPARγ and GLUT4 expression. Normal mice and alloxan-induced diabetic mice were administered fractions of chitin, protein or fat, and the acute hypoglycemic effect was evaluated. A subacute study involving daily administration of these fractions to diabetic mice was also performed over 30 days, after which the liver and pancreas were processed by conventional histological techniques and stained with hematoxylin and eosin to evaluate morphological changes. The most active fraction, the fat fraction, was analyzed by gas chromatography-mass spectrometry (GC-MS), and PPARγ and GLUT4 mRNA expressions were determined in 3T3-L1 adipocytes. The protein and fat fractions exhibited hypoglycemic effects in the acute as well as in the 30-day study. Only the fat fraction led to elevated insulin levels and reduced glycemia, as well as lower intake of water and food. In the liver, we observed recovery of close hepatic cords in the central lobule vein following treatment with the fat fraction, while in the pancreas there was an increased density and percentage of islets and number of cells per islet, suggesting cellular regeneration. The GC-MS analysis of fat revealed three fatty acids as the major components. Finally, increased expression of PPARγ and GLUT4 was observed in 3T3-L1 adipocytes, indicating an antidiabetic effect.

Aplicação da membrana de látex natural e do extrato da pele de rã (Lithobates catesbiana) em feridas cirúrgicas cutâneas de ratos Wistar / Application of natural latex membrane and extract of frog skin (Lithobates catesbiana) (Shaw, 1802) in cutaneous surgical wounds of Wistar rats

Implantou-se a membrana de látex natural e o extrato da pele de rã individualmente e em conjunto em feridas cutâneas de ratos com o intuito de se avaliar o processo de reparação tecidual e possíveis complicações. Utilizou-se 60 ratos da linhagem Wistar divididos em grupos experimentais: grupo controle (GC), grupo membrana de látex (GM), grupo óleo do extrato da pele de rã (GO) e grupo membrana de látex e óleo de rã (GMO), cada um com 15 animais. Aos três, cinco, sete, 14 e 21 dias de pós-operatório, foi realizada eutanásia dos animais para avaliações macroscópicas e histológicas da região lesionada. Nas feridas dos animais do GM e do GC a cicatrização ocorreu mais cedo, no último tempo de avaliação o sinal de lesão era mínimo, já no GMO, a cicatrização não foi completa e no GO a ferida teve o pior resultado, com presença de crosta no 21º dia. Foram observadas, à microscopia de luz, células normais envolvidas no processo de reparação tecidual e formação de neovasos em todos os grupos.  Conclui-se que em todos os grupos não houve rejeição dos biomateriais testados, todavia o grupo GM proporcionou melhor cicatrização com menos efeitos adversos quando comparada aos demais grupos testados.

In this study, the membrane of the natural latex and extract of the skin frog, isolated and together, where implanted to replacea cutaneous in the mice, with purpose for evaluating the tissue repair process and possible complication. Sixty Wistar rats eredivided in four experimental groups: control group (GC), membrane of the latex group (GM), oil of the frog group (GO) and groupmembrane of latex with oil of the frog group (GMO), each one with 15 animals. The animals were euthanized at three, five, seven,14 and 21 days post-operative, followed by macroscopic and microscopic evaluations of the area of the lesion. In the wounds ofthe GM and GC, healing occurred earlier, in the latter evaluation time the signal was minimal injury, in the GMO healing was notcomplete and the wound GO had the worst outcome with presence of crust on the 21st day. There were observed by microscopic,normal cells involved in the tissue repair process and neovascularization in all groups. We conclude that in all groups there wasno rejection of the biomaterials tested, however the GM group gave better healing with fewer adverse effects when compared to the other groups tested.

Effect of a new remineralizing biomaterial on the color of dental enamel

El objetivo de este trabajo fue describir el efecto de la composición de una sustancia remineralizante (SRM) y de la presión osmótica sobre el color dental mediante espectrofotometría. Se tomaron 104 premolares y molares humanos repartidos aleatoriamente en 2 grupos, cada uno de 52 especímenes. El grupo 1 se trató con la sustancia remineralizante SRM 55 (agente remineralizante 1) mezcla de 50 por ciento - 50 por ciento de mineral de grano fino y otro mineral de grano grueso y el grupo 2 se trató con la sustancia remineralizante SRM 91(agente remineralizante 2) contienen los mismos minerales en proporción 90 por ciento - 10 por ciento. A su vez cada grupo se dividió en 2 subgrupos, cada uno de 26 especímenes que se almacenaron así: Un subgrupo en saliva sintética con presión osmótica isotónica (PI) y el otro con presión osmótica hipotónica (PH). Se tomaron registros iniciales y finales con el espectrofotómetro Vita Easy Shade®. Con las lecturas se calcularon losparámetros de color (L*; a*; b*) y los índices de blanqueamiento (WIC; WIO; W). Los cambios de color (ΔL; ΔA; ΔB; yΔE) y los índices de blanqueamiento se compararon y se trataron todos mediante un análisis descriptivo. Las variables ΔA, ΔL, ΔB, ΔE e índice de blanqueamiento W se trataron con ANOVA y los índices WIC y WIO con un análisis de varianza no paramétrico Kruskal Wallis. Los resultados indican que la combinación A2 (SRM 91 y PI) afectó las variables ΔB y ΔE. La combinación B1 (SRM 55 Y PH) afectó las variables ΔA, ΔB y el índice de blanqueamiento WIO. Solamente SRM 91afectó la variable ΔL. La presión osmótica de la saliva y la sustancia remineralizante afectan el color del esmalte dental...

Characterisation of divergent flavivirus NS3 and NS5 protein sequences detected in Rhipicephalus microplus ticks from Brazil

Transcripts similar to those that encode the nonstructural (NS) proteins NS3 and NS5 from flaviviruses were found in a salivary gland (SG) complementary DNA (cDNA) library from the cattle tick Rhipicephalus microplus. Tick extracts were cultured with cells to enable the isolation of viruses capable of replicating in cultured invertebrate and vertebrate cells. Deep sequencing of the viral RNA isolated from culture supernatants provided the complete coding sequences for the NS3 and NS5 proteins and their molecular characterisation confirmed similarity with the NS3 and NS5 sequences from other flaviviruses. Despite this similarity, phylogenetic analyses revealed that this potentially novel virus may be a highly divergent member of the genus Flavivirus. Interestingly, we detected the divergent NS3 and NS5 sequences in ticks collected from several dairy farms widely distributed throughout three regions of Brazil. This is the first report of flavivirus-like transcripts in R. microplus ticks. This novel virus is a potential arbovirus because it replicated in arthropod and mammalian cells; furthermore, it was detected in a cDNA library from tick SGs and therefore may be present in tick saliva. It is important to determine whether and by what means this potential virus is transmissible and to monitor the virus as a potential emerging tick-borne zoonotic pathogen.

Physiological and biochemical characterization of egg extract of black widow spiders to uncover molecular basis of egg toxicity

BACKGROUND: Black widow spider (L. tredecimguttatus) has toxic components not only in the venomous glands, but also in other parts of the body and its eggs. It is biologically important to investigate the molecular basis of the egg toxicity. RESULTS: In the present work, an aqueous extract was prepared from the eggs of the spider and characterized using multiple physiological and biochemical strategies. Gel electrophoresis and mass spectrometry demonstrated that the eggs are rich in high-molecular-mass proteins and the peptides below 5 kDa. The lyophilized extract of the eggs had a protein content of 34.22% and was shown to have a strong toxicity towards mammals and insects. When applied at a concentration of 0.25 mg/mL, the extract could completely block the neuromuscular transmission in mouse isolated phrenic nerve-hemidiaphragm preparations within 12.0 ± 1.5 min. Using whole-cell patch-clamp technique, the egg extract was demonstrated to be able to inhibit the voltage-activated Na+, K+and Ca2+ currents in rat DRG neurons. In addition, the extract displayed activities of multiple hydrolases. Finally, the molecular basis of the egg toxicity was discussed. CONCLUSIONS: The eggs of black widow spiders are rich in proteinous compounds particularly the high-molecular-mass proteins with different types of biological activity The neurotoxic and other active compounds in the eggs are believed to play important roles in the eggs' toxic actions.

Immune enhancement activities of silk lutein extract from Bombyx mori cocoons

BACKGROUND: Declining immune function poses an important clinical challenge worldwide and supplementation with natural products that possessing immune enhancing properties is a promising approach for preventing or delaying immune function decline. Cocoons from yellow silkworms are a significant source of lutein, and this unexplored silk extract could be a viable alternative source for dietary lutein. This study assessed immunomodulatory activities of the silk lutein extract. Female BALB/c mice orally received lutein, either as silk or marigold extracts (10 or 20 mg/kg daily), or vehicle only (1% tween 80 in PBS pH 7.4) for 4 weeks. Natural killer (NK) cell activity, specific antibody production, lymphocyte subpopulations, mitogen-induced lymphocyte proliferation, and cytokine production were examined. RESULTS: Silk lutein extract increased NK cell activity, and the effect was dose-related whereas marigold lutein extract was ineffective. Silk lutein extract dose-dependently enhanced antibody production in pre-immunized mice but marigold lutein extract had no effect. Feeding with silk lutein extract increased the populations of CD3+ and CD4 + CD3 + cells. Silk lutein extract also stimulated concanavalin A- and lipopolysaccharide-induced proliferations of T and B lymphocytes, respectively. Moreover, silk lutein extract increased IL-2 and IFN-γ production while the effect of marigold lutein extract was undetectable. CONCLUSIONS: Together, silk lutein extract enhanced both innate and adaptive immune functions. This preparation may prove to be an effective supplement for strengthened immunity.

Leishmania amazonensis exhibits phosphatidylserine-dependent procoagulant activity, a process that is counteracted by sandfly saliva

Leishmania parasites expose phosphatidylserine (PS) on their surface, a process that has been associated with regulation of host's immune responses. In this study we demonstrate that PS exposure by metacyclic promastigotes of Leishmania amazonensis favours blood coagulation. L. amazonensis accelerates in vitro coagulation of human plasma. In addition, L. amazonensis supports the assembly of the prothrombinase complex, thus promoting thrombin formation. This process was reversed by annexin V which blocks PS binding sites. During blood meal, Lutzomyia longipalpis sandfly inject saliva in the bite site, which has a series of pharmacologically active compounds that inhibit blood coagulation. Since saliva and parasites are co-injected in the host during natural transmission, we evaluated the anticoagulant properties of sandfly saliva in counteracting the procoagulant activity of L. amazonensis . Lu. longipalpis saliva reverses plasma clotting promoted by promastigotes. It also inhibits thrombin formation by the prothrombinase complex assembled either in phosphatidylcholine (PC)/PS vesicles or in L. amazonensis . Sandfly saliva inhibits factor X activation by the intrinsic tenase complex assembled on PC/PS vesicles and blocks factor Xa catalytic activity. Altogether our results show that metacyclic promastigotes of L. amazonensis are procoagulant due to PS exposure. Notably, this effect is efficiently counteracted by sandfly saliva.

Composición química y capacidad anti-irritante de extractos de cuerpo entero de Ulomoides dermestoides (Coleoptera, Tenebrionidae) / Chemical composition and anti-irritant capacity of whole body extracts of Ulomoides dermestoides (Coleoptera, Tenebrionidae)

Background: In traditional medicine of Central and South America, the tenebrionid beetle Ulomoides dermestoides is used as an a phrodisiac, for the treatment of inflammatory diseases and cancer. Recently was reported cytotoxic and genotoxic properties of non-polar extract of U. dermestoides; also anti-inflammatory and immunomodulatory activity of aqueous whole body extract of beetle was reported, it suggests the existence of components with potential pharmacology use. On the other hand, it is necessary to identify those polar and non-polar extracts of U. dermestoides with anti-irritant properties for the membranes and blood vessels, which will be used in subsequence biological test and clinical assays. Objectives: The purpose of this research was to identify the chemical composition of methanolic and hexanic extracts of U. dermestoides, and to assess their anti-irritant capacity. Methods: The extracts were obtained from adult beetles of U. dermestoides. The chemical composition of the extracts was determined by gas chromatography-mass spectrometry (GC-MS) and the anti-irritant effect of each extract was evaluated by means of a modified assay of irritation of the chorioallantoic membrane (CAM) of fertilized chicken eggs (HET-CAM); the results were expressed as irritation index (IR). Results: Six common compounds were identified in both extracts: limonene, myristic, palmitic, estearic, oleic, and linoleic fatty acids. But in the alone methanolic extract were found: 1-pentadecanol, alpha-pinene, beta-phellandrene and alpha-terpinene, whereas in the hexanic extract were found: 2-methyl-p-benzoquinone, 2,4-dihidroxy-1-ethylbenzene, 2,5-dimethyl-quinone, saturated and unsaturated hydrocarbons and alcohols. The methanolic extract of U. dermestoides showed potential anti-irritant effect in the HET-CAM test (IR = 3.09 ± 0.11), similar to that observed with Nimesulida (IR = 2.05 ± 0.14)...

Estudio del perfil lipídico y tejido cardiovascular en ratas alimentadas con grasa Light / Study of lipid profile and cardiovascular tissue in rats fed with fat Light

OBJETIVO: Determinar los efectos deletéreos de los ácidos grasas constituyentes del aceite y/o grasa denominada comercialmente "Light" sobre el perfil lipídico y tejido cardiovascular de ratas albinas sometidas a dietas con requerimientos normales durante 45 días de experimentación. MATERIAL y METODOS: Se trabajó con 30 ratas machos de 21 días de edad, de raza Wistar, distribuidas en tres grupos. Se les proporcionó dietas con energía y proteínas de acuerdo a sus requerimientos durante 45 días. El primer grupo recibió como fuente de ácidos grasas los provenientes de Aceite Vegetal de Cocina (dieta A), el segundo grupo Aceite Vegetal de Cocina denominada "Light" (dieta B) y el tercer grupo Margarina Vegetal denominada "Light" (dieta C). RESULTADOS: Se registró un incremento de peso en los tres grupos y hubo diferencias significativas en la composición corporal (p<0.05) en el grupo alimentado con Margarina Vegetal "Light" (dieta C) comparados con los otros dos grupos (dietas A y B). En los animales alimentados con ácidos grasas constituyentes de la Margarina "Light" (Dieta C) se encontró histológicamente en las fibras musculares una reducción de los paquetes musculares en relación con los otros grupos de Animales, presentando significancia estadística (p<0.01), de igual forma se presentó Disminución de la capa media (compuesta de músculo liso) y luz de la arteria con significancia estadística (p<0.01). En relación a los componentes del perfil lipídico, en los animales alimentados con la dieta A (Colesterol Total, Triglicéridos, Colesterol HDL, Colesterol LDL y Colesterol VLDL), también se presentaron cambios con significancia estadística (p<0.05). CONCLUSIONES: Ratas alimentadas con dietas que incluían ácidos grasas provenientes de la grasa tipo Margarina Vegetal "Light" (Dieta C), presentaron trastornos en la composición corporal, variaciones en el perfil lipídico, con discreto foco degenerativo y reducción del paquete muscular cardíaco.

OBJECTIVE: To determine the deleterious effect of fatty acids included in commercially called 'Light' oil and/or fats upon the lipid profile and cardiovascular tissue in albino rats who received diet complying with the normal requirement during 45 days. MATERIAL AND METHODS: We carried out the trial in 30 male Wistar rats aged 21 days old. The animals were divided in three groups. They received a diet complying with energy and protein requirements during 45 days. The first group received vegetable cooking oil (diet A) as a source of fatty acids, the second group received 'Light' cooking vegetable oil (diet B), and the third group received 'Light' vegetable margarine (diet C). RESULTS: Weight gain was observed in the three groups, and there were significant differences in the body composition (p<0.05) in the group of animals receiving 'Light' vegetable margarine (diet C), compared with the other two groups (diets A and B).Histological studies in the animals that received fatty acids included in 'Light' margarine (diet C) there was a reduction in muscle bundles compared to the other groups, with statistical significance (p<0.01), and also there was a reduction in the media layer (with smooth muscle) and in the arterial lumen, also with statistical significance (p<0.01). With respect to the lipid profile (total cholesterol, HDL-LDL-, and VLDL-cholesterol), the animals that received diet A also had statistically significant changes (p<0.05). CONCLUSIONS: Rats fed with diets containing fatty acids derived from 'Light' vegetable margarine (diet C) had changes in their body composition, in the lipid profile, and mild degenerative changes and a reduction in cardiac muscle mass.

Castration-resistant prostate cancer: systemic therapy in 2012

Prostate cancer is the most common non-cutaneous neoplasm in the male population worldwide. It is typically diagnosed in its early stages, and the disease exhibits a relatively indolent course in most patients. Despite the curability of localized disease with prostatectomy and radiation therapy, some patients develop metastatic disease and die. Although androgen deprivation is present in the majority of patients with metastatic prostate cancer, a state of androgen resistance eventually develops. Castration-resistant prostate cancer, defined when there is progression of disease despite low levels of testosterone, requires specialized care, and improved communication between medical and urologic oncologists has been identified as a key component in delivering effective therapy. Despite being considered a chemoresistant tumor in the past, the use of a prostate-specific antigen has paved the way for a new generation of trials for castration-resistant prostate cancer. Docetaxel is a life-prolonging chemotherapy that has been established as the standard first-line agent in two phase III clinical trials. Cabazitaxel, a novel taxane with activity in cancer models resistant to paclitaxel and docetaxel, is the only agent that has been compared to a chemotherapy control in a phase III clinical trial as a second-line therapy; it was found to prolong the overall survival of patients with castration-resistant prostate cancer previously treated with docetaxel when compared to mitoxantrone. Other agents used in this setting include abiraterone and sipuleucel-T, and novel therapies are continually being investigated in an attempt to improve the outcome for patients with castration-resistant prostate cancer.

Behavioural responses to human skin extracts and antennal phenotypes of sylvatic first filial generation and long rearing laboratory colony Rhodnius prolixus

Chagas disease is a major public health issue and is mainly spread by Triatominae insects (Hemiptera: Reduviidae). Rhodnius prolixus is the main vector species in Northern South America. Host-seeking behaviour in R. prolixus is mediated by different compounds that are produced by and emanate from the host or microbiota on the host's skin. We tested the behavioural responses of sylvatic first filial generation (F1) and colony insects to extracts of human skin with a dual choice olfactometer. In addition, we compared the antennal phenotypes in both populations. No statistical differences were found between the two populations at the behavioural level. Both showed a preference for face and feet extracts and this effect was abolished for face extracts after treatment with an antibacterial gel. The observation of the antennal phenotype showed that there were differences between both groups in the total length, total surface area and number and density of bristles. However, the number and density of chemoreceptive sensilla (basiconic and thin and thick-walled trichoids) and the total density of sensilla did not show statistically significant differences. These results demonstrate that colony insects, which have only been fed with living hens for the last 30 years, are attracted by human skin extracts in a similar way as F1 sylvatic insects.

Classification of brain tumor extracts by high resolution ¹H MRS using partial least squares discriminant analysis

High resolution proton nuclear magnetic resonance spectroscopy (¹H MRS) can be used to detect biochemical changes in vitro caused by distinct pathologies. It can reveal distinct metabolic profiles of brain tumors although the accurate analysis and classification of different spectra remains a challenge. In this study, the pattern recognition method partial least squares discriminant analysis (PLS-DA) was used to classify 11.7 T ¹H MRS spectra of brain tissue extracts from patients with brain tumors into four classes (high-grade neuroglial, low-grade neuroglial, non-neuroglial, and metastasis) and a group of control brain tissue. PLS-DA revealed 9 metabolites as the most important in group differentiation: γ-aminobutyric acid, acetoacetate, alanine, creatine, glutamate/glutamine, glycine, myo-inositol, N-acetylaspartate, and choline compounds. Leave-one-out cross-validation showed that PLS-DA was efficient in group characterization. The metabolic patterns detected can be explained on the basis of previous multimodal studies of tumor metabolism and are consistent with neoplastic cell abnormalities possibly related to high turnover, resistance to apoptosis, osmotic stress and tumor tendency to use alternative energetic pathways such as glycolysis and ketogenesis.

Amino and fatty acid compositions in Haruan traditional extract (HTE) / Composición de aminoácidos y ácidos grasos en extractos de Haruan tradicional

Evaluation of amino and fatty acids compositions in Haruan Traditional Extracts (HTE) was done using HPLC and GC methods. The HTE contained at least 17 amino acids with glutamic acid, glycine, leusine, aspartic acid, proline, alanine and arginine are the most, with values 1.87 - 43.13 mg/g, 21.80 - 80.85 mg/g, 7.85- 40.19 mg/g, 13.85 - 44.07 mg/g, 9.49 - 45.46 mg/g, 11.38 - 35.25 mg/g and 5.99 - 21.79 mg/g, respectively. Meanwhile, the highest percentage of fatty acids is palmitic acid; 3.54 - 26.84 percent of total protein. The others major fatty acids are stearic acid, oleic acid and linoleic acid with values 3.25 - 15.90 percent, 1.40 - 27.68 percent, 0.51 - 7.82 percent of total protein, respectively. HTE also found to have 4 extra bioactive compounds labelled as 1 to 4 on chromatographic tracing which in line with previously finding. It is concluded that the HTE is containing all the important amino acids plus some fatty acids, which is the basis to conduct antioxidant composition in both fresh Haruan and the HTE which was claimed to have wound healing properties. Comparative study was also carried out in various other extraction protocols, including commercial product.

Evaluación de las composiciones de aminoácidos y ácidos grasos en Haruan Extractos tradicional (HTE) se realizó mediante métodos de HPLC y GC. La HTE contenía al menos 17 aminoácidos con ácido glutámico, glicina, leucina, ácido aspártico, prolina, alanina y arginina como mayoritarios, con valores de 1.87 - 43.13 mg/g, 21.80 - 80.85 mg/g, 7.85 - 40.19 mg/g, 13.85 - 44.07 mg/g, 9.49 - 45.46 mg/g, 11.38 - 35.25 mg/g and 5.99 - 21.79 mg/g, respectivamente. Mientras tanto, el mayor porcentaje de ácidos grasos es el ácido palmítico; 3.54 - 26.84 por ciento de la proteína total. Otros ácidos grasos importantes son el ácido esteárico, ácido oleico y ácido linoleico con valores de 3.25 - 15.90 por ciento, 1.40 - 27.68 %, 0.51 - 7.82 por ciento de la proteína total, respectivamente. HTE también encontró cuatro compuestos bioactivos adicionales etiquetados de 1 a 4 en el seguimiento cromatográfico que está de acuerdo con resultados previos. Se concluye que la HTE contiene todos los aminoácidos importantes además de algunos ácidos grasos, que es la base para llevar a cabo la composición antioxidante, tanto en fresco Haruan y la HTE que se afirma poseen propiedades curativas. Estudios comparativos se llevaron a cabo con otros protocolos de extracción, incluido el producto comercial.

Desenvolvimento e validação de método analítico em CLAE-UV para a quantificação de ácido retinóico em microcápsulas de alginato e quitosana / Development and validation of analytical method on HPLC-UV for quantification of retinoic acid in microcapsules of alginate and chitosan

O ácido retinóico (AR) tem sido utilizado para o tratamento de acne severa, rugas, estrias e celulite, no entanto, provoca irritação na pele e sofre rápida degradação quando exposto à luz e ao calor. Métodos analíticos rápidos para quantificação do AR são, portanto, necessários para ensaios de cinética de liberação in vitro. Nesse contexto, o objetivo deste trabalho foi desenvolver e validar um método rápido e sensível para o doseamento do AR em microcápsulas de alginato/quitosana contendo óleo de babaçu dispersas em gel natrosol® por cromatografia líquida de alta eficiência associada à espectroscopia UV e aplicá-lo na avaliação do perfil de liberação in vitro dessas formulações. As análises foram realizadas em modo isocrático utilizando coluna C18 de fase reversa 150 x 4,6 mm (5 μm) com detecção a 350 nm. A fase móvel foi constituída de metanol e ácido acético 1 por cento (85:15 v/v) com vazão de 1,8 mL/minuto. A faixa de linearidade do método foi de 0,5 a 60 μg/mL (r² = 0,999). O método validado mostrou-se sensível, específico, exato, preciso, de baixo custo e o tempo de retenção do AR foi de 5,8 ± 0,4 minutos sendo, desta forma, mais rápido do que os relatados na literatura.

Retinoic acid (RA) has been used in the treatment of severe acne, wrinkles and cellulite. However, it induces skin irritation and rapidly suffers degradation under light and high temperate exposure. Rapid analytical methods to quantify retinoic acid are therefore mandatory for in vitro drug release studies. In this framework, the aim of this study was to develop and validate a rapid and responsive method to quantify the RA in microcapsules of chitosan and alginate containing babassu oil dispersed in natrosol® hydrogel using high performance liquid chromatography (HPLC). Furthermore this method was used to quantify in vitro release kinetics of RA from microcapsules. The analyses have been carried through an isocratic HPLC-UV method using a reversed phase 150 x 4.6 mm C18 (5μm) column, a mobile phase constituted of methanol and 1 percent acetic acid (85:15) at a flow rate of 1.8 mL/min and detection at 350 nm. The linearity range was 0.5-60 μg/mL (r² = 0.999). The validated HPLC-UV method was responsive, specific, accurate, precise, and economic and the RA retention time was 5.8 ± 0.4 minutes, being therefore, faster than that previously reported.

Concentración natural de compuestos antimaláricos en artrópodos tropicales (in vitro)

Natural concentration of antimalaric components in Tropical arthropods (in vitro). Alcohol, hexane and dichlorometane extracts of 751 samples of Costa Rican arthropods were studied for the presence of antimalaric components. With Plasmodium berghei we set an in vitro model in which the effect of the extract was determined by staining of the parasites with cresil brilliant blue. Active extracts at concentration of 50 mg or less, were considered positive. Promissory extracts were found in the orders Lepidoptera (24.1%), Coleoptera (32.8%), Hemiptera (38.5%) and Polydesmida (81.3%). Since most of the Lepidoptera samples were in the immature stages, the relation with the host plant was analyzed. Cannaceae, Flacourtiaceae, Crisobalanaceae, Lauraceae, Fagaceae, Ulmaceae, Rosaceae, Asteraceae, Rubiaceae, Lauraceae and Caprifoliaceae were related with the Lepidoptera larvae, and an antimalaric effect has been reported in most of these families. In the orders Polydesmida, Opiliones and Blattodea, the extract from adults also had some important effect, probably because all of them fed on plants. Polydesmida and Opiliones have chemical substances that probably serve as defensive purposes; these chemicals could also have some antiparasitic effect. Therefore, the detection of antimalaric components in arthropod species led to the identification of plants with promissory antimalaric components. Rev. Biol. Trop. 56 (2): 473-485. Epub 2008 June 30.

Extractos alcohólicos, hexánicos y diclorometánicos de 751 muestras de artrópodos fueron estudiados por la presencia de actividad antimalárica. En este trabajo se empleó un modelo murino usando el Plasmodium berghei, modelo que es biológicamente similar a la malaria humana. El estudio fue realizado determinando el efecto del extracto sobre el parásito por la inclusión o no del colorante azul de cresil brillante. Estimando como positivos aquellos extractos cuya actividad antimalárica se mostró en concentraciones no mayores de 50 mg, se encontró que los órdenes más promisorios fueron Lepidoptera (24.1%), Polydesmida (81.3%), Blattodea (25%) y Opiliones, entre otros. Las formas inmaduras de Lepidoptera fueron las más positivas, por lo que se analizaron las plantas hospederos de donde se alimentaban dichos organismos. Las familias de estas plantas eran Malvaceae, Acanthaceae, Rutaceae, Myrtaceae, Solanaceae, Fabaceae, Urticaceae, Anacardiaceae, Rosaceae, Asteraceae, Rubiaceae, Lauraceae y Caprifoliaceae. Especies de casi todas estas familias han sido reportadas con actividad antimalárica. En el caso de los órdenes Polydesmida, Opiliones y Blattodea, cuyas formas adultas presentaron alguna actividad contra P. berghei, encontramos que todos esos grupos se alimentan también de plantas. En el caso de Opiliones sus especies son predadores de lepidópteros, coleópteros, hemípteros fitófagos y otros artrópodos, además de que producen sustancias de defensas tales como alcoholes, cetonas y quinonas, entre otros, todo lo cual podría explicar la actividad encontrada. Algunas especies del Orden Polydesmida, también secretan ciertas sustancias químicas, las cuales podrían tener un efecto antiparasitario. Así, a través de este trabajo en artrópodos hemos llegado a identificar fuentes vegetales potenciales para componentes antimaláricos.

Immunoblot analysis and comparative IgE responses of atopic patients to extracts of the domestic mite Blomia tropicalis

BACKGROUND: The domestic mite Blomia tropicalis is found in subtropical and tropical environments, and its clinical importance as a sensitizing agent in allergic disease is widely accepted. OBJECTIVE: To investigate the IgE reactivity to allergens present in extracts of the domestic mite B. tropicalis, and compare the IgE responses to these allergens by asthmatics, patients with atopic dermatitis and allergic rhinitis, as well as nonatopic controls. METHODS: Extracts from B. tropicalis were used for skin tests. The B. tropicalis specific IgE in the serum were measured using the FAST Plus Test and immunoblot analysis. RESULTS: A total of 199 volunteers participated in the study. The data show that 18 out of 29 polypeptide bands present in extracts of this mite species were recognized by the allergic and control sera. Of these allergens, four showed a high IgE binding frequency and had relative molecular weights of 104, 80, 68 and 14 kDa. The 14 kDa allergen demonstrated the highest IgE binding frequency. CONCLUSION: Sera from atopic patients reacted to more allergens than sera from patients controls. Extracts from pure bodies of B. tropicalis contain one immunodominant and three important allergens. A common characteristic between all of the sera tested was the high degree of serum IgE reactivity observed to the 14 kDa allergen.

Detection of poliovirus type 2 in oysters by using cell culture and RT-PCR

Devido ao hábito alimentar filtrante, os moluscos bivalves são contaminados por vírus presentes em águas contaminadas por esgoto. Os enterovírus são geralmente usados como modelos para a detecção de vírus em moluscos bivalves devido a sua importância em saúde pública. No presente estudo, ostras foram colocadas em aquários de vidro contendo água do mar adicionada de algas unicelulares. Dois tipos de experimentos foram realizados: a) ostras bioacumulando quatro diferentes concentrações de poliovírus: 5 x 104, 2,5 x 104, 5 x 103, 5 x 102 PFU/mL durante 20h; b) tecidos de ostras inoculados diretamente com 6,0 x 105 e 1,0 x 105 PFU/mL. Após a semeadura, os tecidos foram processados por um método de adsorção-eluição-precipitação. Controles positivos foram realizados por inoculação de 6,0 x 105 PFU/mL de poliovírus diretamente nos tecidos processados das ostras. Os extratos teciduais foram testados para presença do vírus por ensaios de placa de lise (PFU), RT-PCR e cultura celular integrada ao PCR (ICC/PCR). Este último consistiu na inoculação das amostras sobre monocamadas de células VERO seguida de RT-PCR do fluido celular infeccioso. No primeiro experimento (ensaio de bioacumulação por 20h), foram detectados até 5 x 103 PFU de poliovírus, após 24 e 48h de replicação nas células. Os ensaios de RT-PCR e ICC/PCR foram capazes de detectar 3 e 0,04 PFU de poliovírus, respectivamente nos ensaios de bioacumulação. Quando os extratos teciduais processados foram semeados, os ensaios de placa de lise demonstraram recuperação de vírus infecciosos em todas as concentrações testadas. Pudemos concluir que partículas viáveis de poliovírus podem ser detectadas em ostras após bioacumulação e que estas técnicas podem ser diretamente aplicadas na detecção de vírus em amostras ambientais.