Avaliação do impacto de poluentes estrogénicos em peixes

Dissertação de Mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Experience of a Single Center in NTBC Use in Management of Hereditary Tyrosinemia Type I in Libya

Background: Hereditary Tyrosinemia type I (HTI) is a metabolic disease caused by deficiency of fumarylacetoacetate hydrolase enzyme. Objectives: This study reports beside its clinical and biochemical presentation, the outcome of NTBC [2- (2-nitro-4-trifloro-methylbenzoyl)-1, 3-cyclohexanedion] treatment of the disease and evaluates its biochemical markers in 16 pediatric Libyan patients. Patients and Methods: The diagnosis was based on presence of high tyrosine levels in blood and succinylacetone in urine. Results: The consanguinity rate was 81.2%, the median age at onset, at diagnosis and at starting treatment were 4.5, 8, and 9.5 months respectively. At presentation hepatomegaly, jaundice, rickets and high gamma glutamyl transferase (GGT) were observed in 87.5% of patients. All patients had extremely high alpha fetoprotein (AFP) and high alkaline phosphatase (ALP) levels. Fifteen patients were treated with NTBC, normalization of PT (Prothrombine time) was achieved in average in 14 days. The other biochemical parameters of liver function (transaminases, GGT, ALP, bilirubin and albumin) took longer to improve and several months to be normalized. Survival rate with NTBC was 86.6%. Patients who started treatment in a median of 3 months post onset observed a fast drop of AFP in 90.6% of patients (P = 0.003). Abnormal liver function and rickets were the common presentations, GGT was an early cholestatic sensitive test. ALP was constantly high even in asymptomatic patients. Conclusions: In HT1 a faster dropping of AFP is a marker of good prognosis.

The Role of Epstein-Barr Virus LMP-1 Immunohistochemical Staining in Childhood Hodgkin Lymphoma

Background: There are a few published studies about prognostic markers of Epstein-B virus (EBV) related to outcomes in pediatric Hodgkin Lymphoma (HL). Objectives: We aimed to investigate the prognostic value and effect of EBV on survival by using biopsy materials in children and adolescents diagnosed with HL. Patients and Methods: EBV LMP-1 expression was examined using immunohistochemical methods in 58 tumor samples. Clinical features, overall survival (OS) and failure free survival time (FFS) were compared between EBV LMP-1 positive and negative patients. Results: In 20 (35%) patients tumors were LMP-1 positive. When compared with patients above 10 years old, EBV LMP-1 was often positive in patients under 10 years old (30% vs. 70%, P = 0.02). In our most cases having B symptoms and advanced stage, EBV positiveness in Hodgkin Reed-Stenberg cells (H-RS) was not a significant determinant for survival (P = 0.78). Half of the past clinical trials in childhood HL reported longer survival rates in EBV LMP-1 positive patients. In some trials similar to our results there was no significant relationship between EBV and prognosis. Conclusions: The reason of diminished EBV positiviness may be related to technical methods such as not using immunohistochemical and in situ hybridization for EBER antigen but in laboratory conditions painting of control tissues with EBV impair this probability. In addition, cases enrolled to our study were living in Istanbul where social and economical factors are improved rather than generally.

Liver injury from ampicillin-induced intestinal microbiota distresses in rats fed carbohydrate- and protein-rich diets

Purpose: To investigate the effect of ampicillin on rat intestinal microflora and liver in the presence of high carbohydrate and protein diets. Methods: Male Wistar albino rats were divided into four groups. The first group served as the control, the second group was treated with ampicillin (50 mg/kg for 3 weeks) and fed with a standard diet, while the third and fourth groups were treated with the same dose of ampicillin and fed with acarbohydrateand protein-rich diets, respectively, to observe the effect of diet on gut flora and liver. Fecal specimens were collected and used for qualitative determination of gut microbiota composition. Serum hepatospecific markers (AST, ALT and ALP) were estimated. The antioxidant status of liver tissues was estimated for GSH, MDA, GST, LDH and vitamin C l, in addition to sodium and potassium. Results: Administration of orogastric dose of ampicillin for 3 weeks induced inhibition of E.coli, yeasts, total anaerobes, and anaerobic lactobacilli with new growth of P. vulgaris and K. pneumonia. The levels of serum AST, ALT and ALP showed significant (p ˂ 0.05) increase to 163, 112.38 and 115.35 %, respectively in ampicillin-treated animals, compared to control. Also significant (p ˂ 0.05) increase in lipid peroxidation (120 %) and LDH (111 %) coupled with significant (p ˂ 0.05) decrease in glutathione (74.57 %), vitamin C (63.49 %) and glutathione S-transferase (41.51 %) were observed in ampicillintreated groups. No significant variation (p ˂ 0.05) in sodium and potassium levels were found between control and the treated group after 3 weeks of treatment. Conclusion: These results confirm that extended ampicillin therapy disrupts gut flora, which results in liver injury; hence, overuse of antibiotics should be avoid.

X-ray fluorescence spectroscopic determination of heavy metals and trace elements in aerial parts of Origanum sipyleum L from Turkey

Purpose: To determine the heavy metal and trace element composition of the powdered aerial parts of Origanum sipyleum L. and its water extract. Methods: The heavy metal and trace elements content of the powdered plant material and 2 % aqueous extract were evaluated by x-ray fluorescence spectroscopy with silicon drift detector SDD at a resolution of 145 eV and 10,000 pulses. The process conditions were 0.1 g sample weight, process time of 300 s at a voltage of 25 kV and 50 kV, and at a current of 0.5 and 1.0 mA under helium atmosphere. Results: The major elements, K, Ca and Na, known as macronutrients, constituted 11990, 10490 and 970 ppm of the powdered drug and 8910, 2991 and 810 ppm of the water extract, respectively. Among other constituents, arsenic, lead and uranium levels were < 1, 2.1 and < 3 ppm, respectively, in the powdered material while in the aqueous extract, the levels were < 1, < 2 and 200 ppm, respectively. Conclusion: O. sipyleum is a potential source of macro- and micronutrients from which useful food additives and health supplements can be derived.

Investigaci?n del mercado de vidrio y aluminio como materiales de construcci?n y remodelaci?n en el Cant?n Central de Lim?n, que permita a la empresa Inversiones Garita S.A. determinar la posible apertura de una nueva divisi?n de materias primas

Universidad de Costa Rica. Posgrado en Administraci?n y Direcci?n de Empresas. Maestr?a Profesional en Administraci?n y Direcci?n de Empresas, 2015

Protective effect of Rhizoma drynariae extract on osteoporosis in ovariectomized rat model

Purpose: To investigate the therapeutic effect of Rhizoma drynariae extract (RDE) on ovariectomyinduced osteoporosis in rats. Methods: Female Sprague-Dawley rats were randomly assigned to a sham-operated group (control) and five ovariectomy (OVX) subgroups: OVX with vehicle (OVX), OVX with 17ß-estradiol (E2, 25 μg/kg/day), and OVX with RDE doses (40, 80, and 160 mg/kg/day). Daily oral administration of E2 or RDE started 4 weeks after OVX and lasted for 16 weeks. The bone mineral density (BMD) of the L4 vertebrae and right femurs was estimated. The length of each femur was measured with a micrometer gauge, and the center of the diaphysis determined. Three representatives L4 vertebrae were selected to evaluate the trabecular microarchitecture. Serum alkaline phosphatase (ALP), urinary calcium (U-Ca), urinary phosphorus (U-P), urinary creatinine (Cr) and osteocalcin (OC) levels were measured. Results: The study showed that high-dose of RDE significantly inhibited the bone mineral density (BMD) reduction of L4 vertebrae (0.20 ± 0.02 g/cm3, p < 0.05) and femurs (0.18 ± 0.02 g/cm3, p < 0.05) caused by OVX and prevented the deterioration of trabecular microarchitecture (p < 0.05), which were accompanied by a significant decrease in skeletal remodeling (p < 0.05) as evidenced by the lower levels of bone turnover markers. High-dose of RDE improved morphometric parameters, namely, Tb-N (3.8 ± 0.2 mm, p < 0.05), Tb-Th (0.083 ± 0.011 mm, p < 0.05) and Tb-Sp (0.19 ± 0.01 mm, p < 0.05) in L4 vertebrae significantly. The present study indicates that the administration of RDE at higher doses over a 16-week period can prevent OVX-induced osteoporosis in rats without hyperplastic effects on the uterus. Conclusion: Thus, RDE is a potential natural alternative for postmenopausal osteoporosis treatment in elderly women.

Development of hydroxyapatite-based nanomaterials to enhance biological response of osteoblast cells for clinical application

Purpose: To develop a novel chitosan/gelatin-hydroxyapatite (CGHaP) microspheres for evaluating the biological response of pre-osteoblast cells. Methods: The microsphere was prepared by water-in-oil emulsion method. Cell proliferation was studied using AlamarBlue colorimetric assay and DAPI staining while alkaline phosphatase assay was carried out by colorimetric assay method. Chitosan microspheres as well as chitosan-hydroxyapatite microspheres was prepared and tested for biological response from MC3T3-E1 cell line. Results: The results showed that CGHaP promotes MC3T3-E1 cell proliferation and spread on the surface of microspheres. The cells were clustered with more actin filaments and well-linked with neighbouring cells or adjacent cells when cultured in CGHaP microspheres whereas fewer cells were spread on chitosan (CH) microspheres. CGHaP microspheres significantly (p < 0.05) promoted cell attachment, proliferation and extracellular matrix mineralization. CGHaP microspheres presented significantly (p < 0.02) higher calcium deposition (0.5 ng) than CH microspheres (0.28 ng). Specifically, CGHaP microspheres exhibited high ALP activity (8 units; 2-fold) compared to CH with 3 units, after 7 days of incubation. The results suggest that CGHaP possesses a great ability to facilitate bone ingrowth formation and possibility of good osteointegration in vivo. Conclusion: The nanomaterial enhances the proliferation of pre-osteoblast cells in tissue engineering microspheres. The outcome of this study may have a major impact on the development of novel nanomaterials for bone tissue engineering.

Anti-osteoporosis activity of Astragalus membranaceus Bunge extract in experimental rats

Purpose: To investigate the anti-osteoporosis effect of Astragalus membranaceus (Fisch.) Bunge. extract (AMBE) in experimental rats. Method: Female Sprague-Dawley rats were randomly divided into six groups: control group, ovariectomy (OVX) with vehicle group, OVX with 17β-estradiol (E2, 25 μg/kg/day) group, and OVX with AMBE doses (60, 120 and 240 mg/kg/day) groups. Daily oral administration of AMBE or E2 was started 4 weeks after OVX and lasted for 16 weeks. The bone mineral density (BMD) of L4 vertebrae and right femurs was evaluated. The length of each femur was measured with a micrometer, and the center of diaphysis was determined. Three representative L4 vertebrae were selected to evaluate trabecular microarchitecture. Serum alkaline phosphatase (ALP), urinary calcium (U-Ca), urinary phosphorus (UP), urinary creatinine (Cr) and osteocalcin (OC) levels were measured. Results: AMBE dose-dependently inhibited the bone mineral density (BMD) reduction of L4 vertebrae (0.27 ± 0.03 g/cm2, p < 0.05) and femurs (0.23 ± 0.03 g/cm2, p < 0.05) caused by OVX and prevented the deterioration of trabecular microarchitecture (p < 0.05), which were accompanied by a significant decrease in skeletal remodeling (p < 0.05) as evidenced by the lower levels of bone turnover markers. A higher dosage of AMBE treatment (240 mg/kg/day) increased U-Ca/Cr (0.27 ± 0.03 mmol/mmol), ALP (137.23 ± 16.72 U/L), U-P/Cr (4.18 ± 0.27 mmol/mmol) and OC (8.47 ± 0.26 mmol/L) levels (both p < 0.05). Conclusion: The findings of this study indicate that AMBE prevents OVX-induced osteoporosis in rats.

Metais, ambiente e vida: Interferência dovanádio nos sistemas vivos e sua contextualização no ensino das ciências experimentais

A actividade aqui descrita teve como objectivo aproximar alunos do ensino secundário a uma abordagem fisiológica e molecular sobre os efeitos dos metais nos seres vivos. Os alunos ficaram familiarizados com abordagens teóricas e experimentais que permitiram avaliar o crescimento celular por observação qualitativa, determinação do conteúdo proteico e da actividade específica fosfatase alcalina (ALP), recorrendo a centrifugação diferencial, leituras de absorvência e tratamento informático dos resultados. Os estudantes concluíram que a presença de metavanadato de amónio inibiu o crescimento da levedura vínica Saccharomyces cerevisiae UE-ME3, causando um decréscimo significativo do conteúdo proteico e da actividade específica ALP, um marcador da proliferação celular. A avaliação de competências adquiridas, bem como, os inquéritos de opinião mostraram que a maior parte dos alunos do 10° e 12° ano da Escola Secundária Dom Manuel Martins de Setúbal, envolvidos nesta acção, atingiram com sucesso e satisfação os objectivos previamente traçados, revelando uma aprendizagem significativa. ABSTRACT: The work described here aimed to bring high school students to a physiological and molecular approach on the effects of metals in living organisms. The students were familiar with theoretical and experimental approaches that allowed them to evaluate cell growth by qualitative observation, protein contents and alkaline phosphatase (ALP) determination, using differential centrifugation, absorbance readings and computational analysis of data. The students concluded that the presence of ammonium metavanadate inhibited the wine-wild yeast Saccharomyces cerevisiae UE-ME3 growth, and cause a significant decrease of protein content and ALP specific activity, a cell proliferation marker. The assessment of acquired skills, as well as opinion surveys showed that the vast majority of students of 10th and 12nd years of High School Dom Manuel Martins of Setúbal, involved in this project have reached with success and satisfaction the goals previously set for this action and reveals a significant learning.

Biomarker evaluation and toxicological study of essential oils of Foeniculum vulgare and Calamintha nepeta

The aim of this study was to evaluate the toxicological properties of EOs of F. vulgare and C. nepeta, widespread in Mediterranean agrosilvopastoral systems and often used as food condiments in Alentejo. EOs were obtained from aerial part of plants by hydrodistillation and chemical composition was evaluated by GC-FID. Toxicity of essential oils was evaluated by the estimation of LC50 in brine shrimp and LD50 in mice. Oral toxicity assays were performed in mice. Histological analyses and quantification of biomarkers aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma glutamyltransferase (γ-GT), bilirubin, creatinine and urea were performed for monitoring liver and kidney functions. the EOs of C. nepeta and F. vulgare showed very low toxicity suggesting their potential use as food supplement. Additionally, our studies point out the importance of the integration of C. nepeta and F. vugare in silvopastoral agroforestry systems, contributing to the animal health and profitability of livestock.