Search results for: Aurelio Pedroza Sandoval
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 35

Search results for: Aurelio Pedroza Sandoval

5 Comparison of Two Methods of Cryopreservation of Testicular Tissue from Prepubertal Lambs

Authors: Rensson Homero Celiz Ygnacio, Marco Aurélio Schiavo Novaes, Lucy Vanessa Sulca Ñaupas, Ana Paula Ribeiro Rodrigues

Abstract:

The cryopreservation of testicular tissue emerges as an alternative for the preservation of the reproductive potential of individuals who still cannot produce sperm; however, they will undergo treatments that may affect their fertility (e.g., chemotherapy). Therefore, the present work aims to compare two cryopreservation methods (slow freezing and vitrification) in testicular tissue of prepubertal lambs. For that, to obtain the testicular tissue, the animals were castrated and the testicles were collected immediately in a physiological solution supplemented with antibiotics. In the laboratory, the testis was split into small pieces. The total size of the testicular fragments was 3×3x1 mm³ and was placed in a dish contained in Minimum Essential Medium (MEM-HEPES). The fragments were distributed randomly into non-cryopreserved (fresh control), slow freezing (SF), and vitrified. To SF procedures, two fragments from a given male were then placed in a 2,0 mL cryogenic vial containing 1,0 mL MEM-HEPES supplemented with 20% fetal bovine serum (FBS) and 20% dimethylsulfoxide (DMSO). Tubes were placed into a Mr. Frosty™ Freezing container with isopropyl alcohol and transferred to a -80°C freezer for overnight storage. On the next day, each tube was plunged into liquid nitrogen (NL). For vitrification, the ovarian tissue cryosystem (OTC) device was used. Testicular fragments were placed in the OTC device and exposed to the first vitrification solution composed of MEM-HEPES supplemented with 10 mg/mL Bovine Serum Albumin (BSA), 0.25 M sucrose, 10% Ethylene glycol (EG), 10% DMSO and 150 μM alpha-lipoic acid for four min. The VS1 was discarded and then the fragments were submerged into a second vitrification solution (VS2) containing the same composition of VS1 but 20% EG and 20% DMSO. VS2 was then discarded and each OTC device containing up to four testicular fragments was closed and immersed in NL. After the storage period, the fragments were removed from the NL, kept at room temperature for one min and then immersed at 37 °C in a water bath for 30 s. Samples were warmed by sequentially immersing in solutions of MEM-HEPES supplemented with 3 mg/mL BSA and decreasing concentrations of sucrose. Hematoxylin-eosin staining to analyze the tissue architecture was used. The score scale used was from 0 to 3, classified with a score 0 representing normal morphologically, and 3 were considered a lot of alteration. The histomorphological evaluation of the testicular tissue shows that when evaluating the nuclear alteration (distinction of nucleoli and condensation of nuclei), there are no differences when using slow freezing with respect to the control. However, vitrification presents greater damage (p <0.05). On the other hand, when evaluating the epithelial alteration, we observed that the freezing showed scores statistically equal to the control in variables such as retraction of the basement membrane, formation of gaps and organization of the peritubular cells. The results of the study demonstrated that cryopreservation using the slow freezing method is an excellent tool for the preservation of pubertal testicular tissue.

Keywords: cryopreservation, slow freezing, vitrification, testicular tissue, lambs

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4 Peers' Alterity in Inverted Inclusion: A Case Study

Authors: Johanna Sagner, María José Sandoval

Abstract:

At the early stages of adolescence, young people, regardless of a disability or not, start to establish closer friendship ties. Unlike previous developmental phases, these ties are rather reciprocal, more committed, and require more time. Friendship ties during adolescence allow the development of social and personal skills, specifically the skills to start constructing identity. In an inclusive context that incorporates young people with a disability, friendship among peers also takes place. Nonetheless, the relation is shaped, among others, by the alterity construction about the other with disability. Research about peers’ relation between young people with and without disability in an inclusive context has shown that the relation tends to become a helper-helpee relation, where those with a disability are seen as people in need. Prejudices about the others’ condition or distancing from the other because of his/hers disability are common. In this sense, the helper-helpee relation, as a non-reciprocal and protective relation, will not promote friendship between classmates, but a rather asymmetric alterity. Our research is an explorative case study that wants to know how the relation between peers is shaped within a different inclusive program, were also the integrated group has special educational needs. Therefore, we analyze from a qualitative and quantitative approach the data of an inverted inclusive program. This is a unique case of a special public school for visual disability in Germany that includes young people from a mainstream school who had learning difficulties. For the research, we analyze data from interviews, focal interviews and open-ended questions with an interpretative phenomenological analysis approach. The questionnaires include a five point Likert scale, for which we calculate the acceptance rate. The findings show that the alterity relation between pupils is less asymmetrical and represents a rather horizontal alterity. The helper-helpee relation is marked by exchange, since both groups have special educational needs and therefore, those with visual disability and those with learning difficulties help each other indistinctly. Friendship is more present among classmates. The horizontal alterity peers’ relation is influenced by a sort of tie, where none of the groups need more or less help than other groups. Both groups identify that they themselves and the other have special needs. The axiological axe of alterity is not of superiority or inferiority, recognizing each other’s differences and otherness. Another influential factor relates with the amount of time they spend together, since the program does not have a resource room or a teacher who teaches parallel lessons. Two probable causes for that rather equal peer relation might be the constellation of fewer pupils per classroom and the differentiated lessons taught by teachers with a special educational formation.

Keywords: alterity, disability, inverted inclusion, peers’ relation

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3 Sustainability from Ecocity to Ecocampus: An Exploratory Study on Spanish Universities' Water Management

Authors: Leyla A. Sandoval Hamón, Fernando Casani

Abstract:

Sustainability has been integrated into the cities’ agenda due to the impact that they generate. The dimensions of greater proliferation of sustainability, which are taken as a reference, are economic, social and environmental. Thus, the decisions of management of the sustainable cities search a balance between these dimensions in order to provide environment-friendly alternatives. In this context, urban models (where water consumption, energy consumption, waste production, among others) that have emerged in harmony with the environment, are known as Ecocity. A similar model, but on a smaller scale, is ‘Ecocampus’ that is developed in universities (considered ‘small cities’ due to its complex structure). So, sustainable practices are being implemented in the management of university campus activities, following different relevant lines of work. The universities have a strategic role in society, and their activities can strengthen policies, strategies, and measures of sustainability, both internal and external to the organization. Because of their mission in knowledge creation and transfer, these institutions can promote and disseminate more advanced activities in sustainability. This model replica also implies challenges in the sustainable management of water, energy, waste, transportation, among others, inside the campus. The challenge that this paper focuses on is the water management, taking into account that the universities consume big amounts of this resource. The purpose of this paper is to analyze the sustainability experience, with emphasis on water management, of two different campuses belonging to two different Spanish universities - one urban campus in a historic city and the other a suburban campus in the outskirts of a large city. Both universities are in the top hundred of international rankings of sustainable universities. The methodology adopts a qualitative method based on the technique of in-depth interviews and focus-group discussions with administrative and academic staff of the ‘Ecocampus’ offices, the organizational units for sustainability management, from the two Spanish universities. The hypotheses indicate that sustainable policies in terms of water management are best in campuses without big green spaces and where the buildings are built or rebuilt with modern style. The sustainability efforts of the university are independent of the kind of (urban – suburban) campus but an important aspect to improve is the degree of awareness of the university community about water scarcity. In general, the paper suggests that higher institutions adapt their sustainability policies depending on the location and features of the campus and their engagement with the water conservation. Many Spanish universities have proposed policies, good practices, and measures of sustainability. In fact, some offices or centers of Ecocampus have been founded. The originality of this study is to learn from the different experiences of sustainability policies of universities.

Keywords: ecocampus, ecocity, sustainability, water management

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2 Stereological and Morphometric Evaluation of Wound Healing Burns Treated with Ulmo Honey (Eucryphia cordifolia) Unsupplemented and Supplemented with Ascorbic Acid in Guinea Pig (Cavia porcellus)

Authors: Carolina Schencke, Cristian Sandoval, Belgica Vasquez, Mariano Del Sol

Abstract:

Introduction: In a burn injury, the successful repair requires not only the participation of various cells, such as granulocytes and fibroblasts, but also of collagen, which plays a crucial role as a structural and regulatory molecule of scar tissue. Since honey and ascorbic acid have presented a great therapeutic potential to cellular and structural level, experimental studies have proposed its combination in the treatment of wounds. Aim: To evaluate stereological and morphometric parameters of healing wounds, caused by burns, treated with honey Ulmo (Eucryphia cordifolia) unsupplemented, comparing its effect with Ulmo honey supplemented with ascorbic acid. Materials and Methods: Fifteen healthy adult guinea pigs (Cavia porcellus) were used, of both sexes, average weight 450 g from the Centro de Excelencia en Estudios Morfológicos y Quirúrgicos (CEMyQ) at the Universidad de La Frontera, Chile. The animals were divided at random into three groups: positive control (C+), honey only (H) and supplemented honey (SH) and were fed on pellets supplemented with ascorbic acid and water ad libitum, under ambient conditions controlled for temperature, ambient noise and a cycle of 12h light–darkness. The protocol for the experiment was approved by the Scientific Ethics Committee of the Universidad de La Frontera, Chile. The parameters measured were number density per area (NA), volume density (VV), and surface density (SV) of fibroblast; NA and VV of polymorphonuclear cells (PMN) and, evaluation of the content of collagen fibers in the scar dermis. One-way ANOVA was used for statistics analysis and its respective Post hoc tests. Results: The ANOVA analysis for NA, VV and SV of fibroblasts, NA and VV of PMN, and evaluation of collagen content, type I and III, showed that at least one group differs from other (P≤ 0.001). There were differences (P= 0.000) in NA of fibroblast between the groups [C+= 3599.560 mm-2 (SD= 764.461), H= 3355.336 mm-2 (SD= 699.443) and SH= 4253.025 mm-2 (SD= 1041.751)]. The VV and SV of fibroblast increased (P= 0.000) in the SH group [20.400% (SD= 5.897) and 100.876 mm2/mm3 (SD= 29.431), respectively], compared to the C+ [16.324% (SD= 7.719) and 81.676 mm2/mm3 (SD= 28.884), respectively). The mean values of NA and VV of PMN were higher (P= 0.000) in the H [756.875 mm-2 (SD= 516.489) and 2.686% (SD= 2.380), respectively) group. Regarding to the evaluation of the content of collagen fibers, type I and III, the one-way analysis of ANOVA showed a statistically significant difference (P< 0.05). The content of collagen fibers type I was higher in C+ (1988.292 μm2; SD= 1312.379), while the content of collagen fibers type III was higher in SH (1967.163 μm2; SD= 1047.944 μm2) group. Conclusions: The stereological results were correlated with the stage of healing observed for each group. These results suggest that the combination of honey with ascorbic acid potentiate the healing effect, where both participated synergistically.

Keywords: ascorbic acid, morphometry, stereology, Ulmo honey

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1 Effects of Delphinidin on Lipid Metabolism in HepG2 Cells and Diet-Induced Obese Mice

Authors: Marcela Parra-Vargas, Ana Sandoval-Rodriguez, Roberto Rodriguez-Echevarria, Jose Dominguez-Rosales, Juan Armendariz-Borunda

Abstract:

Non-alcoholic fatty liver disease (NAFLD) is characterized by an excess of hepatic lipids, and it is to author’s best knowledge, the most prevalent chronic liver disorder. Anthocyanin-rich food consumption is linked to health benefits in metabolic disorders associated with obesity and NAFLD, although the precise functional role of anthocyanidin delphinidin (Dp) has yet to be established. The aim of this study was to investigate the effect of the Dp in NAFLD metabolic alterations by evaluating prevention or amelioration of hepatic lipid accumulation, as well as molecular mechanisms in two experimental obesity-related models of NALFD. In vitro: HepG2 cells were incubated with sodium palmitate (PA, 1 mM) to induce lipotoxic damage, and concomitantly treated with Dp (180 uM) for 24 h. Subsequently, total lipid accumulation was measured by colorimetric staining with Oil Red O, and total intrahepatic triglycerides were determined by an enzymatic assay. To assess molecular mechanisms, cells were pre-treated with PA for 24 h and then exposed to Dp for 1 h. In vivo: four-week-old male C57BL/6Nhsd mice were allocated in two main groups. Mice were fed with standard diet (control) or high-fat and high-carbohydrate diet (45% fat, HFD) for 16 wk to induce NAFLD. Then HFD was divided into subgroups: one treated orally with Dp (15 mg/kg bw, HFD-Dp) every day for 4 wk, while HFD group treated with vehicle (DMSO). Weight and fasting glucose were recorded weekly, while dietary ingestion was measured daily. Insulin tolerance test was performed at the end of treatment. Liver histology was evaluated with H&E and Masson’s trichrome stain. RT-PCR was used to evaluate gene expression and Western Blot to determine levels of protein in both experimental models. Parametric data were analyzed with one-way ANOVA and Tukey’s post-hoc test. Kruskal-Wallis and Mann-Whitney U test for non-parametric data, and P < 0.5 were considered significant. Dp prevented hepatic lipid accumulation by PA in HepG2 hepatocytes. Furthermore, Dp down-regulated gene expression of SREBP1c, FAS, and CPT1a without modifying AMPK phosphorylation levels. In vivo, Dp oral administration did not ameliorate lipid metabolic alterations raised by HFD. Adiposity, dietary ingestion, fasting glucose, and insulin sensitivity after Dp treatment remained similar to HFD group. Histological analysis showed hepatic damage in HFD groups and no differences between HFD and HFD-Dp groups were found. Hepatic gene expression of ACC and FAS were not altered by HFD. SREBP1c was similar in both HFD and HFD-Dp groups. No significant changes were observed in SREBP1c, ACC, and FAS adipose tissue gene expression by HFD or Dp treatment. Additionally, immunoblotting analysis revealed no changes in pathway SIRT1-LKB-AMPK and PPAR alpha by both HFD groups compared to control. In conclusion, the antioxidant Dp may provoke beneficial effects in the prevention of hepatic lipid accumulation. Nevertheless, the oral dose administrated in mice that simulated the total intake of anthocyanins consumed daily by humans has no effect as a treatment on hepatic lipid metabolic alterations and histological abnormalities associated with exposure to chronic HFD. A healthy lifestyle with regular intake of antioxidants such as anthocyanins may prevent metabolic alterations in NAFLD.

Keywords: anthocyanins, antioxidants, delphinidin, non-alcoholic fatty liver disease, obesity

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