Search results for: Michaela Quintero

Authors: Michaela Storen, Michelle Harvey, Xavier Conlan

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Gun violence internationally is increasing at an unprecedented level, becoming a favoured means for executing violence against another individual. Not only is this putting a strain on forensic scientists who attempt to determine the cause of death in circumstances where firearms have been involved in the death of an individual, but it also highlights the need for an alternative technique of identification of a gunshot wound when other established techniques have been exhausted. A corpse may be colonized by necrophagous insects following death, and this close association between the time of death and insect colonization makes entomological samples valuable evidence when remains become decomposed beyond toxicological utility. Entomotoxicology provides the potential for the identification of toxins in a decomposing corpse, with recent research uncovering the capabilities of entomotoxicology to detect gunshot residue (GSR) in a corpse. However, shortcomings of the limited literature available on this topic have not been addressed, with the bioaccumulation, detection limits, and sensitivity to gunshots not considered thus far, leaving questions as to the applicability of this new technique in the forensic context. Larvae were placed on meat contaminated with GSR at different concentrations and compared to a control meat sample to establish the uptake of GSR by the larvae, with bioaccumulation established by placing the larvae on fresh, uncontaminated meat for a period of time before analysis using ICP-MS. The findings of Pb, Ba, and Sb at each stage of the lifecycle and bioaccumulation in the larvae will be presented. In addition, throughout these previously mentioned experiments, larvae were washed once, twice and three times to evaluate the effectiveness of existing entomological practices in removing external toxins from specimens prior to entomotoxicologyical analysis. Analysis of these larval washes will be presented. By addressing these points, this research extends the utility of entomotoxicology in cause-of-death investigations and provides an additional source of evidence for forensic scientists in the circumstances involving a gunshot wound on a corpse, in addition to advising the effectiveness of current entomology collection protocols.

Keywords: bioaccumulation, chemistry, entomology, gunshot residue, toxicology

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Authors: Petra Borilova Linhartova, Michaela Ruckova, Sabina Sevcikova, Natalie Mlcuchova, Jan Bohm, Katerina Zukalova, Monika Vlachova, Jiri Dolina, Lumir Kunovsky, Radek Kroupa, Zdenek Pavlovsky, Zdenek Danek, Tereza Deissova, Lydie Izakovicova Holla, Ondrej Slaby, Zdenek Kala

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Esophageal adenocarcinoma (EAC) is a highly aggressive malignancy that frequently develops from Barrett's esophagus (BE), a premalignant pathologic change occurring in the lower end of the esophagus. Specific microRNAs (miRNAs), small non-coding RNAs that function as posttranscriptional regulators of gene expression, were repeatedly proved to play key roles in the pathogenesis of these diseases. This pilot study aimed to analyze four selected miRNAs in esophageal tissues from healthy controls (HC) and patients with reflux esophagitis (RE)/BE/EAC, as well as to compare expression at the site of Barrett's mucosa/adenocarcinoma and healthy esophageal tissue outside the area of the main pathology in patients with BE/EAC. In this pilot study, 22 individuals (3 HC, 8 RE, 5 BE, 6 EAC) were included and endoscopically examined. RNA was isolated from the fresh-frozen esophageal tissue (stored in the RNAlater™ Stabilization Solution −70°C) using the AllPrep DNA/RNA/miRNA Universal Kit. Subsequent RT-qPCR analysis was performed using selected TaqMan MicroRNA Assays for miR-21, miR-34a, miR-196a, miR-196b, and endogenous control (RNU44). While the expression of miR-21 in the esophageal tissue with the main pathology was decreased in BE and EAC patients in comparison to the group of HC and RE patients (p=0.01), the expression of miR-196a was increased in the BE and EAC patients (p<0.01). Correlations between those miRNAs expression in tissue and severity of diagnosis were observed (p<0.05). In addition, miR-196a was significantly more expressed at the site with the main pathology than in paired adjacent esophageal tissue in BE and EAC patients (p<0.01). In conclusion, our pilot results showed that miR-196a, which regulates the proliferation, invasion, and migration (and was previously associated with esophageal squamous cell carcinoma and marked as a potential therapeutic target), could be a diagnostic tissue biomarker for BE and EAC as well.

Keywords: microRNA, barrett´s esophagus, esophageal adenocarcinoma, biomarker

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Authors: Jason Clarke, Michaela Porubanova, Angela Mazzoli, Gulsah Kut

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What accounts for our perception that time inexorably passes in one direction, from the past to the future, the so-called arrow of time, given that the laws of physics permit motion in one temporal direction to also happen in the reverse temporal direction? Modern physics says that the reason for time’s unidirectional physical arrow is the relationship between time and entropy, the degree of disorder in the universe, which is evolving from low entropy (high order; thermal disequilibrium) toward high entropy (high disorder; thermal equilibrium), the second law of thermodynamics. Accordingly, our perception of the direction of time, from past to future, is believed to emanate as a result of the natural evolution of entropy from low to high, with low entropy defining our notion of ‘before’ and high entropy defining our notion of ‘after’. Here we explored this proposed relationship between entropy and the perception of time’s arrow. We predicted that if the brain has some mechanism for detecting entropy, whose output feeds into processes involved in constructing our perception of the direction of time, presentation of violations to the expectation that low entropy defines ‘before’ and high entropy defines ‘after’ would alert this mechanism, leading to measurable behavioral effects, namely a disruption in duration perception. To test this hypothesis, participants were shown briefly-presented (1000 ms or 500 ms) computer-generated visual dynamic events: novel 3D shapes that were seen either to evolve from whole figures into parts (low to high entropy condition) or were seen in the reverse direction: parts that coalesced into whole figures (high to low entropy condition). On each trial, participants were instructed to reproduce the duration of their visual experience of the stimulus by pressing and releasing the space bar. To ensure that attention was being deployed to the stimuli, a secondary task was to report the direction of the visual event (forward or reverse motion). Participants completed 60 trials. As predicted, we found that duration reproduction was significantly longer for the high to low entropy condition compared to the low to high entropy condition (p=.03). This preliminary data suggests the presence of a neural mechanism that detects entropy, which is used by other processes to construct our perception of the direction of time or time’s arrow.

Keywords: time perception, entropy, temporal illusions, duration perception

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Authors: Eimar Yadir Quintero Tapias

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Background: Science is not generally self-correcting; many clinical studies end with the same conclusion "more research is needed." This study hypothesizes that first, we need a better appraisal of the available (and unavailable) evidence instead of creating more of the same false inquiries. Methods: Systematic review of ClinicalTrials.gov study records using the following Boolean operators: (food OR nut OR milk OR egg OR shellfish OR wheat OR peanuts) AND (allergy OR allergies OR hypersensitivity OR hypersensitivities). Variables included the status of the study (e g., active and completed), availability of results, sponsor type, sample size, among others. To determine the rates of non-publication in journals indexed by PubMed, an advanced search query using the specific Number of Clinical Trials (e.g., NCT000001 OR NCT000002 OR...) was performed. As a prophylactic measure to prevent P-hacking, data analyses only included descriptive statistics and not inferential approaches. Results: A total of 2092 study records matched the search query described above (date: September 13, 2019). Most studies were interventional (n = 1770; 84.6%) and the remainder observational (n = 322; 15.4%). Universities, hospitals, and research centers sponsored over half of these investigations (n = 1208; 57.7%), 308 studies (14.7%) were industry-funded, and 147 received NIH grants; the remaining studies got mixed sponsorship. Regarding completed studies (n = 1156; 55.2%), 248 (21.5%) have results available at the registry site, and 417 (36.1%) matched NCT numbers of journal papers indexed by PubMed. Conclusions: The internal and external validity of human research is critical for the appraisal of medical evidence. It is imperative to analyze the entire dataset of clinical studies, preferably at a patient-level anonymized raw data, before rushing to conclusions with insufficient and inadequate information. Publication bias and non-registration of clinical trials limit the evaluation of the evidence concerning therapeutic interventions for food allergy, such as oral and sublingual immunotherapy, as well as any other medical condition. Over half of the food allergy human research remains unpublished.

Keywords: allergy, clinical trials, immunology, systematic reviews

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Authors: Alvarado M. Lady G., Ortiz V. Yaylenne, Quintero B. Quelbis R.

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The oil palm sector has seen significant growth in Colombia after the insertion of policies to stimulate the use of biofuels, which eventually contributes to the reduction of greenhouse gases (GHG) that deteriorate not only the environment but the health of people. However, the policy of using biofuels has been strongly questioned by the impacts that can generate; an example is the increase of other more harmful GHGs like the CH₄ that underlies the amount of solid waste generated. Casanare's department is estimated be one of the major producers of palm oil of the country given that has recently expanded its sowed area, which implies an increase in waste generated primarily in the industrial stage. For this reason, the following study evaluated the agronomic potential of the biochar obtained from empty fruit bunches and its nutritional contribution in cultivated soils with Elaeis guineensis in Casanare, Colombia. The biochar was obtained by slow pyrolysis of the clusters in a retort oven at an average temperature of 190 °C and a residence time of 8 hours. The final product was taken to the laboratory for its physical and chemical analysis as well as a soil sample from a cultivation of Elaeis guineensis located in Tauramena-Casanare. With the results obtained plus the bibliographical reports of the nutrient demand in this cultivation, the possible nutritional contribution of the biochar was determined. It is estimated that the cultivation requirements of nitrogen is 12.1 kg.ha⁻¹, potassium is 59.3 kg.ha⁻¹, magnesium is -31.5 kg.ha⁻¹ and phosphorus is 5.6 kg.ha⁻¹ obtaining a biochar contribution of 143.1 kg.ha⁻¹, 1204.5 kg.ha⁻¹, 39.2 kg.ha⁻¹ and 71.6 kg.ha⁻¹ respectively. The incorporation of biochar into the soil would significantly improve the concentrations of N, P, K and Mg, nutrients considered important in the yield of palm oil, coupled with the importance of nutrient recycling in agricultural production systems sustainable. The biochar application improves the physical properties of soils, mainly in the humidity retention. On the other hand, it regulates the availability of nutrients for plants absorption, with economic savings in the application of synthetic fertilizers and water by irrigation. It also becomes an alternative to manage agricultural waste, reducing the involuntary emissions of greenhouse gases to the environment by decomposition in the field, reducing the CO₂ content in the atmosphere.

Keywords: biochar, nutrient recycling, oil palm, pyrolysis

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Authors: Carlos A. Garcia-Mogollon, Juan C. Quintero-Diaz, Claudio Avignone-Rossa

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Clostridium saccharoperbutylacetonicum 1-4N (Csb 1-4N) is an industrial reference strain for Acetone-Butanol-Ethanol (ABE) fermentation. Csb 1-4N is a solventogenic clostridium and H₂ producer with a metabolic profile that makes it a good candidate for Bioelectrosynthesis System (BES). The aim of this study was to evaluate the electroactivity of Csb 1-4N by cyclic voltammetry technique (CV). The Bioelectrosynthesis fermentation (BES) started in a Triptone-Yeast extract (TY) medium with trace elements and vitamins, Complex Nitrogen Source (CNS), and bicarbonate (NaHCO₃, 4g/L) as a carbon source, run at -600mVAg/AgCl and adding 200uM NADH. The six BES batches were performed with different media composition with and without NADH, CNS, HCO₃⁻ , and applied potential. The CV was performed as three-electrode system: platinum slice working electrode (WE), nickel contra electrode (CE) and reference electrode Ag/AgCl (ER). CVs were run in a potential range of -0.7V to 0.7V vs. VAg/AgCl at a scan rate 10mV/s. A CV recorded using different NaHCO₃ concentrations (0.25; 0.5; 1.0; 4g/L) were obtained. BES fermentation samples were centrifuged (3000 rpm, 5min, 4C), and supernatant (7mL) was used. CVs were obtained for Csb1-4N BES culture cell-free supernatant at 0h, 24h, and 48h. The electrochemical analysis was carried out with a PalmSens 4.0 potentiostat/galvanostat controlled with the PStrace 5.7 software, and CVs curves were characterized by reduction and oxidation currents and reduction and oxidation peaks. The CVs obtained for NaHCO₃ solutions showed that the reduction current and oxidation current decreased as the NaHCO₃ concentration was decreased. All reduction and oxidation currents decreased until exponential growth stop (24h), independence of initial cathodic current, except in medium with trace elements, vitamins, and NaHCO3, in which reduction current was around half at 24h and followed decreasing at 48. In this medium, Csb1-4N did not grow, but pH was increased, indicating that NaHCO₃ was reduced as the reduction current decreased. In general, at 48h reduction currents did not present important changes between different mediums in BES cultures. In terms of intensities in the peaks (Ip) did not present important variations; except with Ipa and Ipc in BES culture with NaHCO₃ and NADH added are higher than peaks in other cultures. Based on results, cathodic and anodic currents changes were induced by NaHCO₃ reduction reactions during Csb1-4N metabolic activity in different BES experiments.

Keywords: clostridium saccharoperbutylacetonicum 1-4N, bioelectrosynthesis, carbon dioxide fixation, cyclic voltammetry

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Authors: Homyr L. Garcia, Jr., Anne Margaret A. Rendon, Carla Michaela B. Taguinod

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Nothing is constant in this world but change. This is the reality wherein a lot of people fail to recognize and maybe, it is because of the fact that some see things that are happening with little value or no value at all until it’s gone. For the past years, Egypt was known for its stable government. It was able to withstand a lot of problems and crisis which challenged their country in ways which can never be imagined. In the present time, it seems like in just a snap of a finger, the said stability vanished and it was immediately replaced by a crisis which resulted to a failure in some parts of their government. In addition, this problem continued to worsen and the current situation of Egypt is just a reflection or a result of it. On the other hand, as the researchers continued to study the reasons why the government of Egypt is unstable, they concluded that there might be a possibility that they will be able to produce ways in which their country could be helped or improved. The instability of the government of Egypt is the product of combining all the problems which affects the lives of the people. Some of the reasons that the researchers found are the following: 1) unending doubts of the people regarding the ruling capacity of elected presidents, 2) removal of President Mohamed Morsi in position, 3) economic crisis, 4) a lot of protests and revolution happened, 5) resignation of the long term President Hosni Mubarak and 6) the office of the President is most likely available only to the chosen successor. Also, according to previous researches, there are two plausible scenarios for the instability of Egypt: 1) a military intervention specifically the Supreme Council of the Armed Forces or SCAF, resulting from a contested succession and 2) an Islamist push for political power which highlights the claim that religion is a hindrance towards the development of their country and government. From the eight possible reasons, the researchers decided that they will be focusing on economic crisis since the instability is more clearly seen in the country’s economy which directly affects the people and the government itself. In addition, they made a hypothesis which states that stable economy is a prerequisite towards a stable government. If they will be able to show how this claim is true by using the Social Autopsy Research Design for the qualitative method and Pearson’s correlation coefficient for the quantitative method, the researchers might be able to produce a proposal on how Egypt can stabilize their government and avoid such problems. Also, the hypothesis will be based from the Rational Action Theory which is a theory for understanding and modeling social and economy as well as individual behavior.

Keywords: Pearson’s correlation coefficient, rational action theory, social autopsy research design, supreme council of the armed forces (SCAF)

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Authors: Mauricio Quintero Angel, Alexander Pereira, Selene Alarcón

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One of the elements of greatest importance in agricultural production is the harvesting; an activity associated to different occupational health risks such as harvesting in high altitudes, the transport of heavy materials and the application of excessive muscle strain that leads to muscular-bone disorders. Therefore, there is an urgent necessity to improve and validate interventions to reduce exposition and risk to harvesters. This article has the objective of describing the co-development under the ergonomic analysis framework of an assisted manual harvesting tool for peach palm oriented to reduce the risk of death and accidents as it avoid the harvest in heights. The peach palm is a palm tree that is cultivated in Colombia, Perú, Brasil, Costa Rica, among others and that reaches heights of over 20 m, with stipes covered with spines. The fruits are drupes of variable size. For the harvesting of peach palm, in Colombia farmers use the “Marota” or “Climber”, a tool in a closed X shape built in wood, that has two supports adjusted at the stipe, that elevate alternately until reaching a point high enough to grab the bunch that is brought down using a rope. An activity of high risk since it is done at a high altitude without any type of protection and safety measures. The Marota is alternated with a rod, which as variable height between 5 and 12 Meters with a harness system at one end to hold the bunch that is lowered with the whole system (bamboo bunch). The rod is used from the ground or from the Marota in height. As an alternative to traditional tools, the Bajachonta was co-developed with farmers, a tool that employs a traditional bamboo hook system with modifications, to be able to hold it with a rope that passes through a pulley. Once the bunch is hitched, the hook system is detached and this stays attached to the peduncle of the palm tree, afterwards through a pulling force being exerted towards the ground by tensioning the rope, the bunch comes loose to be taken down using a rope and the pulley system to the ground, reducing the risk and efforts in the operation. The bajachonta was evaluated in tree productive zones of Colombia, with innovative farmers, were the adoption is highly probable, with some modifications to improve its efficiency and effectiveness, keeping in mind that the farmers perceive in it an advantage in the reduction of death and accidents by not having to harvest in heights.

Keywords: assisted harvesting, ergonomics, harvesting in high altitudes, participative design, peach palm

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Authors: Genevieve Jorolan Quintero

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In Mindanao, the southern island of the Philippines, two provinces, Davao Oriental and Tboli of South Cotabato, respectively, are homes to indigenous communities known for their dream weavers. Davao Oriental is home to the Mandaya, while Lake Sebu is home to the Tboli. The dream weavers are mostly women who have continued the tradition of weaving, a spiritual practice of handicraft embodying the beliefs of the community. It is believed that a weaver is guided by the Tagamaling, or the nature spirit in Mandaya mythology, and Fu Dalu, or the spirit of the abaca among the Tboli. In the dream, the Tagamaling or Fu Dalu reveals to the weaver the design or the pattern of the dagmay as the abaca woven cloth is called among the Mandaya and the tnalak among the Tboli. The weaver then undertakes the production of this nature-spirit-inspired fabric based on her memory of the dream. This interaction between the spirit world and the human world inspired the theme of the short story with the title Loom of Dreams, published in 2015 by Kritika Kultura, an international peer-reviewed journal of language and literary/cultural studies of the Ateneo de Manila University in the Philippines. In Lake Sebu, a collection of the legendary tnalak with various designs is preserved by the cultural advocate and tnalak collector Reden S. Ulo. About a hundred tnalak designs are housed in a mini museum. The paper discusses how the dagmay and the tnalak of the two Philippine indigenous communities, the Mandaya and the Tboli, embody their folklore and cultural heritage. The specific objectives are: 1. To describe the role of the dreamweavers among the Mandaya and Tboli communities in the Philippines; 2. To analyse how folklore influences the designs on the woven fabric, the dagmay, and the tnalak, and 3. To discuss how dream-weaving helps preserve culture legacy. Ethnography was used in the conduct of this research. Specifically, the following data collection methods were done: 1. a series of visits to the Mandaya and Tboli communities; 2. face-to-face interviews with the respondents from the communities, and 3. the recording of the interviews with the knowledge-bearers and material culture keepers from both communities, the narratives of which were used as a basis for the data analysis. The influence of folklore in the culture and the arts of the indigenous communities is significantly evident in the designs of the dagmay and the tnalak. As the dream weavers continue to weave the dagmay and the tnalak, this cultural legacy will continue to prosper and be preserved for posterity.

Keywords: dreamweaver's, Mandaya, mindanao, Philippine folklore, Tboli

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Authors: Guillermo E. Quintero, Edwin G. Perez, Oriel Sanchez, Christian Espinosa-Bustos, Denis Fuentealba, Margarita E. Aliaga

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Bisulfite ion (HSO3-) has been used as a preservative in food, drinks, and medication. However, it is well-known that HSO3- can cause health problems like asthma and allergic reactions in people. Due to the above, the development of analytical methods for detecting this ion has gained great interest. In line with the above, the current use of colorimetric and/or fluorescent probes as a detection technique has acquired great relevance due to their high sensitivity and accuracy. In this context, 2-quinolinone derivatives have been found to possess promising activity as antiviral agents, sensitizers in solar cells, antifungals, antioxidants, and sensors. In particular, 7-(diethylamino)-2-quinolinone derivatives have attracted attention in recent years since their suitable photophysical properties become promising fluorescent probes. In Addition, there is evidence that photophysical properties and reactivity can be affected by the study medium, such as micellar media. Based on the above background, 7-(diethylamino)-2-quinolinone derivatives based chalcone will be able to be incorporated into a cationic micellar environment (Cetyltrimethylammonium bromide, CTAB). Furthermore, the supramolecular control induced by the micellar environment will increase the reactivity of these derivatives towards nucleophilic analytes such as HSO3- (Michael-type addition reaction), leading to the generation of new colorimetric and/or fluorescent probes. In the present study, two derivatives of 7-(diethylamino)-2-quinolinone based chalcone DQD1-2 were synthesized according to the method reported by the literature. These derivatives were structurally characterized by 1H, 13C NMR, and HRMS-ESI. In addition, UV-VIS and fluorescence studies determined absorption bands near 450 nm, emission bands near 600 nm, fluorescence quantum yields near 0.01, and fluorescence lifetimes of 5 ps. In line with the foregoing, these photophysical properties aforementioned were improved in the presence of a cationic micellar medium using CTAB thanks to the formation of adducts presenting association constants of the order of 2,5x105 M-1, increasing the quantum yields to 0.12 and the fluorescence lifetimes corresponding to two lifetimes near to 120 and 400 ps for DQD1 and DQD2. Besides, thanks to the presence of the micellar medium, the reactivity of these derivatives with nucleophilic analytes, such as HSO3-, was increased. This was achieved through kinetic studies, which demonstrated an increase in the bimolecular rate constants in the presence of a micellar medium. Finally, probe DQD1 was chosen as the best sensor since it was assessed to detect HSO3- with excellent results.

Keywords: bisulfite detection, cationic micelle, colorimetric probes, quinolinone derivatives

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Authors: Genevieve Jorolan-Quintero

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The recording as well as the preservation of indigenous literature is an important task as it deals with a significant heritage of pre-colonial culture. The beliefs and traditions of a people are reflected in their oral narratives, such as the folk epic, which must be written down to insure their preservation. The epic poem for instance, known as dawot among the Mandaya, one of the indigenous communities in the southern region of the Philippines, narrates the customs, the ways of life, and the adventures of an ancient people. Nabayra, an expert on Philippine folkloric studies, stresses that still extant after centuries of unknown origin, the dawot was handed down to the magdadawot (bard) by word of mouth, forming the greatest bulk of Mandaya oral tradition. Unhampered by modern means of communication to distract her/him, the magdadawot has a sharp memory of the intricacies of the ancient art of chanting the panayday (verses) of the epic poem. The dawot has several hullubaton (episodes), each of which takes several nights to chant . The language used in these oral traditions is archaic Mandaya, no longer spoken or clearly understood by the present generation. There is urgency to the task of recording and writing down what remain of the epic poem since the singers and storytellers who have retained the memory and the skill of chanting and narrating the dawot and other forms of oral tradition in their original forms are getting fewer. The few who are gifted and skilled to transmit these ancient arts and wisdom are old and dying. Unlike the other Philippine epics (i.e. the Darangen, the Ulahingan, the Hinilawod, etc.), the Mandaya epic is yet to be recognized and given its rightful place among the recorded epics in Philippine Folk Literature. The general aim of this study was to put together and preserve an intangible heritage, the Mandaya hullubaton (episodes of the dawot), in order to preserve and promote appreciation for the oral traditions and cultural legacy of the Mandaya. It was able to record, transcribe, and translate four hullubaton of the folk epic into two languages, Visayan and English to insure understanding of their contents and significance among non-Mandaya audiences. Evident in the contents of the episodes are the cultural practices, ideals, life values, and traditions of the ancient Mandaya. While the conquests and adventures of the Mandaya heroes Lumungtad, Dilam, and Gambong highlight heroic virtues, the role of the Mandaya matriarch in family affairs is likewise stressed. The recording and the translation of the hullubaton and the dawot into commonly spoken languages will not only promote knowledge and understanding about their culture, but will also stimulate in the members of this cultural community a sense of pride for their literature and culture. Knowledge about indigenous cultural system and philosophy derived from their oral literature will serve as a springboard to further comparative researches dealing with indigenous mores and belief systems among the different tribes in the Philippines, in Asia, in Africa, and other countries in the world.

Keywords: Dawot, epic poem, Mandaya, Philippine folk literature

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Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

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Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

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