Beslenme ve Diyetetik Bölümü Koleksiyonu

Permanent URI for this collectionhttps://hdl.handle.net/20.500.12514/223

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Antioxidant, Ache Inhibitory, and Anticancer Effects of Verbascum Thapsus Extract
(C M B Assoc, 2023) Zhang, Na; Baran, Ayse; Valioglu, Ferzane; Teng, Lei; Atalar, Mehmet Nuri; Keskin, Cumali; Beilerli, Aferin
Verbascum thapsus (Mullein) is a medicinal plant used in folk medicine to treat various ailments. For this study, the biological functions of Verbascum thapsus (VT) methanol extract were determined in vitro. The plant's methanol extract was created through the maceration process. The phytochemical composition of plant extracts was investigated using liquid chromatography-electrospray ionization tandem mass spectrometry. The antioxidant capacity of the extract was determined using the 2,2-diphenyl-1-picrylhydrazil (DPPH radical) and 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS radical). Cell lines Caco-2 (human colorectal adenocarcinoma cells), LNCaP (Lymph Node Carcinoma of Prostate), and HEK293 (Human embryonic kidney 293 cells) were used to model colon, prostate, and non-cancerous cells. The cytotoxic activity of the plant extract on the proliferation of these cells was determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole) assay protocol. VT extract showed moderate DPPH and ABTS radical scavenging activities at 30 mg/ml concentration. With this, VT extract was determined to inhibit acetylcholinesterase (AChE) enzyme and had strong cytotoxic activity on cancerous cell lines. In addition, our findings clearly showed that the plant extract had greater cytotoxic activity on the viability of cancerous cells compared to non-cancerous (Human embryonic kidney cells; HEK293) cells. The current findings showed that V. thapsus might be a promising anti-cancer medication candidate for the treatment of human colorectal adenocarcinoma and colon cancer, as well as a good source antioxidans. Copyright: (c) 2023 by the C.M.B. Association. All rights reserved.
Assessment of renal function in obese and overweight children with ngal and kim-1 biomarkers
(Nutricion Hospitalaria, 2020) Unsal, Velid; Gül, Ali; Yılmaz, Resul; Özmen, Zeliha Cansel; Demir, Osman; Gümüşer, Rüveyda
Aim and background:the incidence of obesity has increased among children, and obesity has been considered an independent risk factor for chronic kidney disease. We aimed to determine the degree of kidney function impairment by evaluating urine neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) levels. Materials and methods: in total, 15 obese, 26 overweight, and 26 control adolescents aged 10 to 16 years were enrolled into the study. Urine samples were evaluated for NGAL and KIM-1 levels using enzyme-linked immunosorbent assay kits. We investigated the association between obesity and related comorbidities with urinary NGAL and KIM-1 excretion. Results: no significant differences were noted between the obese, overweight, and control groups in urinary NGAL and KIM-1 excretion (p = 0.327 and p = 0.917, respectively). In the obese and overweight groups urinary NGAL levels were 50.39 [30.88-74.22] in females and 26.67 [23.24-45.59] in males (p = 0.013). Also, urinary NGAL levels were increased in obese and overweight adolescents with LDL dyslipidemia at 64.12 [30.98-114.32] as compared to those without LDL dyslipidemia: 39.51 [25.59.56.37] (p = 0.024). Furthermore, a correlation was observed between insulin and homeostasis model assessment of insulin resistance levels with the NGAL/creatinine ratio in the overweight group (r = 0.515; p = 0.008, and r = 0.483; p = 0.014, respectively). Such correlation was not found in the obese group. Conclusion: the effect of obesity on renal function could not be determined in children. A longer exposure may be required for obesity-induced disruption of renal function in children. Renal function may be disrupted by dyslipidemia in obese adolescents. Furthermore, obesity impaired renal function in female adolescents. The normalization of these urinary markers as related to urine creatinine should be discussed.
Effect of seasons on the fatty acid profile of total lipid, phospholipid and triacylglycerol in Mastacembelus mastacembelus (Atatürk Dam Lake, Turkey).
(2018) Kaçar, Semra; Başhan, Mehmet; Oymak, Ahmet
The present study investigated the lipid content and FA profiles of total lipids, phospholipids (PL) and triacylglycerols (TAG) of Mastacembelus mastacembelus from the Atatürk Dam Lake (Turkey). The results showed that the total lipid content of the dorsal muscle varied seasonally from 0.50% to 3.59%. Wide ranges of saturated fatty acids (SFAs) (36.21 to 50.52%) and polyunsaturated fatty acids (PUFAs) (25.22 to 42.02%) were found in the PL fraction. However, higher monounsaturated fatty acids (MUFAs) (37.63 to 45.07%) and SFA (34.71 to 38.82%) were found in TAG (triacylglycerol). The ratios of ω-3 to ω-6 PUFA ranged from 0.65 to 1.32 and 1.07 to 3.48 in PL (phospholipid) and TAG fraction, respectively. The results also showed that the major components were palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1ω-9), palmitoleic acid (C16:1ω-7), docosahexaenoic acid (DHA, C22:6ω-3), arachidonic acid (AA, C20:4ω-6) and docosapentaenoic acid (C22:5ω-3) in total lipid, C16:0, C18:1ω-9, AA and DHA in PL, C16:0, C16:1ω-7, C18:1ω-9, linoleic acid (LA, C18:2ω-6), and DHA in TAG extracted from the muscle of M. mastacembelus in all seasons.
Effects of ketamine on penile tissues in an experimental priapism model in rats
(Ali Cangül, 2024) Kölükçü, Vildan; Gürler Balta, Mehtap; Tapar, Hakan; Karaman, Tuğba; Karaman, Serkan; Ünsal, Velid; Gevrek, Fikret; Yalçın, Kenan; Fırat, Fatih
Background: This study aimed to evaluate the histopathological and biochemical effects of ketamine on penile tissues following ischemia-reperfusion injury induced by priapism. Methods: Twenty-four male rats were randomized into three groups. Group 1 served as the control group. Group 2 underwent the priapism model to induce ischemia-reperfusion injury. Group 3, the treatment group, experienced a similar ischemia-reperfusion model as Group 2; additionally, 50 mg/kg of ketamine was administered intraperitoneally just before reperfusion. Blood biochemical analyses and penile histopathological evaluations were performed. Results: In Group 3, significant improvements were observed in all histopathological scores, including desquamation, edema, inflammation, and vasocongestion compared to Group 2 (p<0.001). Blood biochemical analyses showed that the malondialdehyde (MDA) levels were recorded as 10 in Group 2, with a significant decrease in Group 3 (p=0.013). Similarly, proinflammatory cytokine levels, including interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α), were found to be suppressed in Group 3 compared to Group 2 (p=0.003, p=0.022, and p=0.028, respectively). Antioxidant enzyme activities, such as glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD), were higher in Group 3 compared to Group 2 (p=0.016 and p=0.024, respec-tively). Conclusion: Ketamine is an effective anesthetic agent in alleviating the effects of penile ischemia-reperfusion injury.
Experimental and theoretical investigation of adsorption and inhibition properties of 2-Amino-1,3,5-triazine-4,6-dithiol against corrosion in hydrochloric acid solution on mild steel
(Elsevier, 2023) Arslanhan, Selim; Yıldız, Reşit; Döner, Ali; Yıldız, Reşit
In this work, 2-Amino-1,3,5-triazine-4,6-dithiol (2-ATD) as novel and high efficiency corrosion inhibitor has been investigated for mild steel (MS) corrosion in 0.5 M HCl solution using electrochemical methods, scanning electron microscopy (SEM), energy disperse X-ray spectroscopy (EDX), atomic force microscopy (AFM) and quantum chemical calculation methods. Potentiodynamic polarization (PDP) curves indicate that 2-ATD is mixed type inhibitor, corrosion inhibition efficiency increased with increasing inhibitor concentration and reached its value of 96.5%. Evolution of exposure time versus corrosion behavior of 2-ATD is examined in corrosive medium. While corrosion potential (Ecorr) shifted more negative values, polarization resistances (Rp) decreased after 120 h exposure time due to the corrosion process. H2 volume is measured in uninhibited and inhibited solutions (10 mM 2-ATD) after 120 h exposure time. Very low volume (3.6 mL cm-2) of H2 is obtained on MS electrode in inhibited solution after 120 h of exposure, indicating that 2-ATD covers the entire surface against aggressive attack and retards the both anodic dissolution of MS and cathodic hydrogen evolution reactions. The adsorption process proposal is the Langmuir isotherm which is most suitable. Adsorption and thermodynamic parameters show that 2-ATD has a strong adsorption effect onto MS surface and includes mixed adsorption style (physical and chemical). Corrosion current density increases with increasing temperature and high activation energy (Ea) proves the strong adsorption of 2-ATD on the MS surface. Anti-corrosion mechanism of 2-ATD is described more detail with the potential of zero charge method. SEM, EDX and AFM analysis support the obtained results of electrochemical methods and confirm the existence of protective layer and strong adsorption of 2-ATD on the MS surface. Chronoamperometry test shows that current densities are almost constant whole experiment in the presence of organic film. Finally, quantum chemical calculation method of 2-ATD in blank solution is performed to investigate the active sites for possible attachment with MS surface.
Experimental and theoretical investigation of pyrazinecarboxamide against mild steel corrosion
(Journal of Dispersion Science and Technology, 2021) Keleşoğlu, Ayşen; Sığırcı, Gökmen; Yıldız, Reşit; Dehri, İlyas; Yıldız, Reşit
Pyrazinecarboxamide was examined against mild steel corrosion in 0.5 M HCl solution. The electrochemical techniques like electrochemical impedance spectroscopy, linear polarization resistance and potentiodynamic polarization measurements were carried out in the presence of various concentrations of pyrazinecarboxamide ranging from 0.25 mM to 1.0 mM. The inhibition effect of pyrazinecarboxamide on the mild steel surface was studied by atomic force microscopy and scanning electron microscopy. The adsorption of this inhibitor was determined to fit Langmuir adsorption isotherm. The adsorption/desorption equilibrium constant was calculated as 2.985 × 104 M−1, indicating that high proportion of pyrazinecarboxamide takes place on mild steel surface. The inhibition efficiency was found as 93.5% from electrochemical impedance spectroscopy measurement in an acid solution containing 1.0 mM pyrazinecarboxamide. Quantum chemical parameters that were in good agreement with experimental results were determined by density functional theory.
Fatty acid compositions of colostrum and mature breast milk in Turkey (Mardin)
(Acta Alimentaria, 2023) Kaçar, Semra; Başhan, Mehmet; Keskin, Cumali
In this study, 25-25 milk samples (25 colostral and 25 mature) collected at different lactational stages were used to analyse the fatty acid composition of breast milk. A gas-chromatographic method was used to perform and analyse the transmethylation of total milk lipid extracts. The milk samples contained 20 different fatty acids. Palmitic acid (C16:0), stearic acid (C18:0), myristic acid (C14:0), oleic acid (C18:1n-9), and linoleic acid (LA, C18:2n-6) were the major components of total lipid, phospholipid (PL), and triacylglycerol (TAG) fractions. Colostrum had a lower percentage of polyunsaturated fatty acids (PUFAs), a higher percentage of saturated fatty acids (SFAs), and a lower level of eicosapentaenoic acid (EPA, C20:5 n-3) and docosahexaenoic acid (DHA, C22:6 n-3) in total lipid than mature milk. Triacylglycerol and PL fractions between colostrum and mature milk samples did not differ statistically. © 2023 Akadémiai Kiadó, Budapest.
Green biosynthesis of silver nanoparticles using Prunus cerasifera pissardii nigra leaf and their antimicrobial activities against some food pathogens
(Czech Journal of Food Sciences, 2022) Hatipoğlu, Abdulkerim
In this study, silver nanoparticles (AgNPs) were synthesised using the Prunus cerasifera pissardii nigra (PC) leaf extract in an easy, low-cost and environmentally friendly way. According to the ultraviolet-visible (UV-vis) spectrophotometer analysis data, the nanocrystals demonstrated a characteristic peak at 456 nm. Scanning electron microscopy (SEM), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM) and energy-dispersive X-ray (EDX) spectroscopy analyses revealed that the morphological structures of the biosynthesised AgNPs were mostly spherical. According to the results of X-ray diffraction (XRD) analysis, it was determined that the crystal structures of AgNPs were cubic. The size of the nanoparticles was calculated as 23.60 nm using the Debye-Scherrer equation. The zeta potential of the synthesised nanomaterial was measured as –15.5 mV. The minimum inhibitory concentration (MIC) values of AgNPs on Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Bacillus subtilis ATCC 11774, Pseudomonas aeruginosa ATCC 27853 and Candida albicans ATCC 10231 were determined to be 0.062, 0.250, 0.125, 0.500 and 0.125 µg mL–1, respectively
Green synthesis of gold nanoparticles from prunus cerasifera pissardii nigra leaf and their antimicrobial activities on some food pathogens
(Progress in Nutrition, 2021) Hatipoğlu, Abdulkerim
In this study, a new and easy method for the biosynthesis of gold nanoparticles (AuNPs) with Prunus cerasifera pissardii nigra (PC) leaf extract as a reducing and stabilizing agent was presented. The nanoparticles were demonstrated a characteristic peak at the maximum wavelength of 535 nm with colour change as a result of the ultraviolet (UV)-visible spectrophotometer analysis data. Emission Scanning Electron Microscopy (FE-SEM), Transmission Electron Microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDX) analyzes revealed that the crystal size of the synthesized AuNPs was below 20 nm and the morphological structure was mostly spherical. The size of the crystal structures of AuNPs was calculated as 17.94 nm from the X-ray diffraction (XRD) analysis data. Fourier Transform Infrared (FT-IR) Spectroscopy results confirmed the involvement of various biomolecules in the reduction and stabilization of PC-AuNPs. The zeta potential of the synthesized nanomaterial was measured as -27 mV. The average size of AuNPs was determined as 103.8 nm with Zetasizer. It was determined that AuNPs have strong inhibitory activity against Escherichia coli, Staphylococcus aureus, Bacillus subtillis and Pseudomonas aeruginosa and Candida albicans.
The interaction methylene blue and glutathione-S-transferase purified from human erythrocytes
(Journal of Planar Chromatography - Modern TLC, 2020) Acay, Hilal; Uzan, Serhat; Baran, Mehmet Fırat; Bilden, Alican; Aygün, Hüsamettin
It is known that textile dyes have various risks on human health. Glutathione-S-transferase enzymes play a critical role in the detoxification of xenobiotics in living systems. This study aimed to examine the interaction of methylene blue with human erythrocyte glutathione-S-transferase purified in one step. Human erythrocyte glutathione-S-transferase was purified with approximately 750-fold purification and 30% efficiency by glutathione agarose affinity chromatography. The results showed that the enzyme was inhibited by methylene blue with an IC50 value of 1.40 mmol/L. The Ki constant of methylene blue was 1.17 mmol/L. The Lineweaver–Burk graph of the methylene blue showed that the type of inhibition was compatible with mixed type inhibition. A new third spot was also detected by thin-layer chromatography. Furthermore, the cytotoxicity of methylene blue on human erythrocytes was evaluated and it was found that the haemolysis per cent of methylene blue on erythrocytes was approximately 14%.
Investigation of the Relationship Cellular and Physiological Degeneration in the Mandible With Aqp1 and Aqp3 Membrane Proteins
(De Gruyter Open Ltd, 2020) Çiçek, M.; Unsal, V.; Tümer, M.K.
Objective: In this study, we aimed to investigate the changes in the levels of oxidative stress and antioxidant enzymes on the mandibular bone caused by the expression of aquaporin-1 and aquaporin-3 proteins. Material and method: 14 Balb/C white mice were divided into two groups of seven, based on whether they are young or old. Mandibular tissue samples were taken for biochemical and histological analysis. Results: Findings of our study has shown that, AQP-1 and AQP-3 immunoreactivity significantly decreased in mandibular bone tissues of aged mice in comparison to younger mice (p < 0.05). MDA and AOPP levels, which are the indicators of oxidative stress, increased in elderly mice and antioxidant defense system SOD enzyme activity was decreased (p < 0.05). The TNF-α cytokine level, which is the indicator for inflammations, was found to be higher in older mice than in young mice (p < 0.05). Conclusion: As a result, it was observed that cellular damage, disruption in water – electrolyte balance and increased inflammation that occur during the natural process of aging had caused serious and irreversible disturbances. © 2020 De Gruyter. All rights reserved.
Protective effects of apocynin and melatonin on ovarian ischemia/reperfusion injury in rats
(Clinical and Experimental Obstetrics and Gynecology, 2021) Unsal, Velid; İşgüder, Çiğdem Kunt; Can, Özlem Koşar; Pektaş, Mine Kanat; Arici, Akgül
Objective: This study aims to determine the protective effects of apocynin, a NADPH oxidase inhibitor, and melatonin, an endogenous anti-oxidant, in an animal model of ovarian ischemia/reperfusion (I/R) injury. Materials/Methods: Thirty-five female rats were randomly divided into five groups, namely group I (sham), group II (I/R), group III (I/R + 10 mg/kg apocynin), group IV (I/R + 20 mg/kg apocynin), and group V (I/R + 10 mg/kg melatonin). Ovarian tissue and serum superoxide dismutase (SOD) and catalase (CAT) activities and malondialdehyde (MDA) and protein carbonyl (PC) levels were measured. Ovarian histopathology was examined and Bax, caspase 3, and iNOS immunoreactivities were evaluated. Results: Preoperative apocynin and melatonin significantly increased SOD and CAT activities (P < 0.05, P < 0.05, P < 0.01, P < 0.01, P < 0.01, P < 0.05, P < 0.05, and P<0.01, respectively, for both apocynin and melatonin). In addition, preoperative apocynin and melatonin significantly decreased the ovarian I/R injury score (P < 0.01 for both). Bax, caspase 3, and iNOS immunoreactivities were significantly lower in the I/R + 10 mg/kg apocynin and I/R + 10 mg/kg melatonin groups than in the I/R group (P<0.01, P<0.01, P<0.01, P<0.01, P<0.05, and P<0.01, respectively). Conclusions: Apocynin and melatonin are powerful antioxidant agents with considerable bioavailability and safety. Preoperative apocynin and melatonin administration might protect ovarian tissue from I/R injury after surgical adnexal detorsion.
The protective effects of sinapic acid on acute renal ischemia/reperfusion injury
(Turkish Journal of Biochemistry, 2021) Unsal, Velid; kolukcu, Engin; Firat, Fatih; Gevrek, Fikret
Objectives: The aim of this research was to investigate whether sinapic acid (SA) can alleviate oxidative damage, apoptosis, and inflammation in I/R induced renal injury. Methods: A total of 24 male rats were randomly separated into four groups as six rats in each group. Group 1 (Sham), Group 2 (I/R), Group 3 (I/R + SA, 10mg/kg), Group 4 (I/R + SA, 20 mg/kg). In order to evaluate kidney function serum BUN, Cr, and AST were measured in an autoanalyzer. SOD, GSH-Px, MDA, PC and NO oxidative stress parameters were measured with spectrophotometric methods and TNF-α, IL-1β, IL-6, KIM-1 and NGAL parameters were measured with the ELISA method. In addition, H&E method and immunohistochemical examinations were performed for histological evaluations of kidney tissue. Results: SA significantly decreases the increase in kidney damage, inflammation, oxidative stress, cell death and restore the decrease in antioxidant enzyme activities (p<0.05). Pre-treatment of the rats with SA reduces kidney dysfunction and morphological changes. Conclusions: The development of oxidative stress and lipid peroxidation seems to be the leading factors that accelerate inflammation and cell death during renal IRI. The antioxidant, anti-inflammatory, and anti-apoptotic features of SA displayed a renoprotective effect.
Rapid green synthesis of gold nanoparticles: synthesis, characterization, and antimicrobial activities
(Progress in Nutrition, 2021) Hatipoğlu, Abdulkerim
In this research, an easy and environmentally friendly method was presented for the biosynthesis of gold nanoparticles (AuNPs) with Allium ampeloprasum (AA) leaf extract as a reducing and stabilizing agent. The maximum absorption peak was found to be around 535 nm with ultraviolet (UV)-visible spectrophotometer. When the Transmission Electron Microscopy (TEM), Emission Scanning Electron Microscopy (FE-SEM) and energy dispersive X-ray (EDX) analyzes were examined, it was seen that the crystal size of the synthesized AuNPs was between 6.02-17.36 nm and their shape was mostly spherical. The size of the crystal structures of AuNPs was calculated as 22.76 nm from the X-ray diffraction (XRD) analysis data. Fourier Transform Infrared (FT-IR) Spectroscopy results showed that phenolics, aromatic compounds and proteins were effective in the reduction and stabilization of AA-AuNPs. The average size of AuNPs was determined as 96.46 nm with Zetasizer. It was determined that AuNPs have a very strong inhibitory effect on pathogenic bacteria (Escherichia coli, Staphylococcus aureus, Bacillus subtillis and Pseudomonas aeruginosa) and Candida albicans by the minimum inhibitory concentration (MIC) method.
Renoprotective effect of diacerein in rats with partial unilateral ureteral obstruction model
(WALTER DE GRUYTER, 2024) Kölükçü, Engin; Unsal, Velid; Fırat, Fatih; Gevrek, Fikret; Katar, Muzaffer
Objectives: We aimed to analyze the effects of diacerein in a rat model of partial unilateral ureteral obstruction (PUUO). Methods: We randomly divided 24 female rats into three groups. Control group, PUUO group and PUUO + diacerein group. The PUUO group was subjected to the PUUO model for seven days. The PUUO + diacerein group received oral diacerein (80 mg/kg) for seven days. Spectrophotometric methods were employed to measure oxidative stress parameters, including malondialdehyde (MDA), protein carbonyl (PC) and antioxidant enzyme levels, including glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD), while indicators of renal function, such as kidney injury molecule-1 (KIM-1) and neutrophil gelatinous-associated lipocalin (NGAL), along with inflammatory parameters interleukin-1 beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6), were assessed using the ELISA method. Inflammatory parameters were measured in blood samples, and other parameters were analyzed in kidney tissue. Hematoxylin-eosin method examinations were used for histological analyses. Results: IL-1beta, TNF-alpha, and IL-6 levels were found to be significantly decreased in the PUUO + diacerein group compared to the PUUO group (p=0.006, p=0.002 and p=0.001, respectively). In the PUUO + diacerein group, GSH-Px and SOD activities increased compared with the PUUO group (p=0.031 and p=0.037, respectively). We also observed a significant improvement in renal function parameters, such as KIM-1 and NGAL levels in the PUUO + diacerein group compared to PUUO (p=0.002 and p=0.012, respectively). The PC and MDA levels were highest in the PUUO group (p<0.001). Similarly, the histopathologic tissue damage was the most prominent PUUO group (p<0.001). Conclusions: Our study found that diacerein is a highly effective pharmacologic agent in alleviating oxidative damage in PUUO model rats.
Seasonal effects of the fatty acid composition of phospholipid and triacylglycerol in the muscle and liver of male Salmo trutta macrostigma
(GRASAS Y ACEITES, 2021) Kaçar, Semra; Kaya, H.; Başhan, M.
The seasonal effects on the fatty acid composition of triacylglycerol (TAG) and phospholipid (PL) in the muscle and liver of male Salmo trutta macrostigma were determined using the gas chromatographic (GC) method. The fatty acid (FA) compositions of total lipid, PL and TAG fractions were determined in muscle and liver tissues of S. trutta macrostigma. The phospholipids contained a higher proportion of 16:0 compared to the TAG in the muscle tissue of S. trutta macrostigma. Docosahexaenoic acid (22:6 ω-3) and eicosapentaenoic acid (20:5 ω-3) contents were high in both muscle and liver tissues. The total lipid contents in the muscle and liver were 1.07-2.45 and 3.00-4.64%, respectively. S. trutta macrostigma is a rich source of ω-3 and ω-6, polyunsaturated fatty acids (PUFA) with numerous benefits to human health. © 2021 CSIC Consejo Superior de Investigaciones Cientificas. All rights reserved.

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