DEHP exposure impairs human skeletal muscle cell proliferation in primary culture conditions: preliminary study.

The plasticizer di (2-ethylhexyl) phthalate (DEHP) inhibits differentiation, impairs glucose metabolism, and decreases mitochondrial function in murine muscle satellite cells; however, if these effects are translated to human cells is unknown. The goal of this study was to evaluate changes in morphology and proliferation of primary human skeletal muscle cells exposed to DEHP. Rectus abdominis muscle samples were obtained from healthy women undergoing programed cesarean surgery. Skeletal muscle cells were isolated and grown under standard primary culture conditions, generating two independent sample groups of 25 subcultures each. Cells from the first group were exposed to 1 mM DEHP for 13 days and monitored for changes in cell morphology, satellite cell frequency and total cell abundance, while the second group remained untreated (control). Differences between treated and untreated groups were compared using generalized linear mixed models (GLMM). Cell membrane and nuclear envelope boundary alterations, loss of cell volume and presence of stress bodies were observed in DEHP-treated cultures. DEHP-treated cultures also showed a significant reduction in satellite cell frequency compared to controls. Exposure to DEHP reduced human skeletal muscle cell abundance. Statistical differences were found between the GLMM slopes, suggesting that exposure to DEHP reduced growth rate. These results suggest that exposure to DEHP inhibits human skeletal muscle cell proliferation, as evidenced by reduced cell abundance, potentially compromising long-term culture viability. Therefore, DEHP induces human skeletal muscle cell deterioration potentially inducing an inhibitory effect of myogenesis by depleting satellite cells.

Dolphin leukocytes exhibit an attenuated cytokine response and increase heme oxygenase activity upon exposure to lipopolysaccharides.

Cetaceans exhibit physiological adaptations that allowed the transition to aquatic life, including a robust antioxidant defense system that prevents injury from repeated exposure to ischemia/reperfusion events associated with breath-hold diving. The signaling cascades that characterize ischemic inflammation in humans are well characterized. In contrast, cetaceans' molecular and biochemical mechanisms that confer tolerance to inflammatory events are poorly understood. Heme oxygenase (HO) is a cytoprotective protein with anti-inflammatory properties. HO catalyzes the first step in the oxidative degradation of heme. The inducible HO-1 isoform is regulated by various stimuli, including hypoxia, oxidant stress, and inflammatory cytokines. The objective of this study was to compare the response of HO-1 and cytokines to a proinflammatory challenge in leukocytes isolated from humans and bottlenose dolphins (Tursiops truncatus). We measured changes in HO activity, and abundance and expression of interleukin 1 beta (IL-1ß), interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-α), and heme oxygenase 1 (HMOX1) in leukocytes treated with lipopolysaccharide (LPS) for 24 and 48 h. HO activity increased (p < 0.05) in dolphin (48 h) but not human cells. TNF-α expression increased in human (24 h, 48 h), but not dolphin cells following LPS stimulation. LPS-induced cytokine expression was lower in dolphin than in human leukocytes, suggesting a blunted cytokine response in bottlenose dolphin leukocytes treated with LPS. Results suggest species-specific regulation of inflammatory cytokines in leukocytes treated with LPS, which may lead to differential responses to a pro-inflammatory challenge between marine and terrestrial mammals.

Spatial and Temporal Variability of Trace and Macro Elements in the Red Crab Pleuroncodes planipes in the Pacific Coast of the Baja California Peninsula, Mexico.

The red crab, Pleuroncodes planipes, is a decapod crustacean abundant off the Pacific coast of the Baja California Peninsula. This species is caught and used in preparing animal feed, such as flour, particularly for aquaculture. Levels of calcium (Ca), cadmium (Cd), copper (Cu), iron (Fe), lead (Pb), magnesium (Mg), manganese (Mn), nickel (Ni), phosphorus (P), and zinc (Zn) were measured in red crabs collected from three geographic zones during three cruises in different seasons. Significant differences were found in the levels of Ca, Cd, Cu, Fe, Mg, Ni, P, and Zn between the two El Niño years (cruises C1 and C3, based on a threshold of ±0.5 °C for the Oceanic Niño Index). The highest concentrations of most elements were observed in the south of the Baja California Peninsula, a highly productive area influenced by upwelling events. Our findings suggest that while environmental temperature plays a central role in the benthic or pelagic distribution of red crabs, their content and variability of trace and macro elements appear to be associated with the presence of oceanic conditions, such as upwelling and potential changes in the composition of their diet associated with the depth in which these crustaceans are collected.

"Oxidative stress induced by phthalates in mammals: State of the art and potential biomarkers".

BACKGROUND: Phthalates, plasticizers that are widely used in consumer products including toys, cosmetics, and food containers, have negative effects in liver, kidney, brain, lung and reproductive system of humans and other mammals. OBJECTIVES: To summarize, describe and discuss the available information on the effects of phthalate exposure in mammals, with emphasis on oxidative stress, and to suggest potential biomarkers of the health risks associated with phthalate exposure. METHODS: An assessment of scientific journals was performed using the PRISMA model for systematic reviews. Manuscripts reporting effects of phthalate exposure on mammalian health published in the last decade were selected according to originality, content, and association to health hazards. RESULTS AND DISCUSSION: We identified 25 peer-reviewed articles published between January 1st, 2010 and June 1st, 2021 that fit the aims and selection criteria. Phthalates induce oxidative stress and cell degenerative processes by increasing intracellular reactive species. Antioxidant cytoprotective systems decrease with time of exposure; conversely, oxidative damage markers, including thiobarbituric acid-reactive substances (TBARS), 8-hydroxy-2'-desoxyguanosine (8-OHdG) and malondialdehyde (MDA), increase. Phthalates were associated with endocrine system disfunction, metabolic disorders, infertility, nonviable pregnancy, cell degeneration, growth impairment, tumor development, and cognitive disorders. Phthalates can also aggravate health conditions such as asthma, hepatitis, diabetes, allergies, chronic liver and kidney diseases. Among humans, the more vulnerable subjects to phthalate exposure effects were children and individuals with a prior health condition. CONCLUSION: Chronic exposure to phthalates induces oxidative stress in mammals with concomitant adverse effects in reproductive, respiratory, endocrine, circulatory, and central nervous systems in both in vitro and in vivo trials. Oxidative damage markers and phthalate metabolites levels were the most common biomarkers of phthalate exposure effects. Studies in free-ranging and wild mammals are nil. Further studies on the pathways that lead to metabolic disruption are needed to identify potential treatments against phthalate-induced detrimental effects.

Morphological alterations in human skeletal muscle cells exposed to di-(2-Ethylhexyl) Phthalate (dehp) in primary cell culture conditions / Alteraciones morfológicas en células musculares esqueléticas humanas expuestas a Ftalato de di-(2-etilhexilo) (dehp) en condiciones de cultivo celular primario

SUMMARY: Di-(2-ethylhexyl) phthalate (DEHP) is among the most common plasticizer additives that humans are in contact with daily. DEHP can be released from plastic and enter the human body, whereby it is metabolized and transformed into oxidative hydrophilic molecules. Clinical follow-ups in patients exposed to this phthalate and investigations in cultures of several cell types have provided information on its effects. For example, it is associated with inhibition of diploid human cell development and morphological changes in cultured germ cells. Although skeletal muscle represents around 50 % of the human body mass, knowledge about the effects of DEHP on this tissue is poor. Cultured skeletal muscle cells were exposed to DEHP (1 mM) for 13 days with the aim of exploring and evaluating some of the potential morphological effects. Three culture development parameters and nine cell characteristics were monitored during the bioassay. At 13 days, growth area, cell viability, and concentration of total proteins were lower in DEHP exposed than in control cells. Cell width and area, as well as the diameter of the nucleus and nucleolus, were greater in exposed cells than in control cells. These are interpreted as signs of cytotoxicity and suggest potential adverse effects on the development of skeletal muscle cells from DEHP exposure, as reported for other cell types.

RESUMEN: Diariamente los seres humanos tenemos contacto con aditivos plastificantes, el di-(2-etilhexil) ftalato (DEHP) se encuentra entre los más comunes. El DEHP puede liberarse del plástico e ingresar al cuerpo humano, donde es metabolizado y transformando en moléculas hidrofílicas oxidativas. Seguimientos en pacientes expuestos a este ftalato e investigaciones en cultivos de varios tipos celulares han aportado información sobre sus efectos. El DEHP es asociado con la inhibición del desarrollo de células humanas diploides y cambios morfológicos en células germinales en cultivo. Sin embargo, aún es poco lo que se sabe sobre los efectos en el músculo esquelético, a pesar de que este tejido representa alrededor del 50 % de la masa corporal del humano. Para explorar y evaluar algunos efectos morfológicos en células de músculo esquelético, cultivos primarios fueron expuestos a DEHP (1 mM) durante 13 días. Se dio seguimiento a tres parámetros de desarrollo del cultivo y nueve características celulares. Al término de 13 días de exposición, los valores del área de crecimiento, viabilidad celular y concentración de proteínas totales fueron inferiores con respecto a los cultivos control. Se observaron cambios morfométricos en las células expuestas. Particularmente, el ancho y área celular, así como los diámetros del núcleo y nucleolos, fueron mayores a los registros en las células control. Estos resultados se interpretan como signos de citotoxicidad y sugieren efectos potencialmente adversos en el desarrollo de las células del músculo esquelético ante una exposición al DEHP, como se ha registrado para otros tipos celulares.

Morphometric characteristics of human skeletal muscle cells in primary culture / Características morfométricas de células musculares esqueléticas humanas en cultivo primario

SUMMARY: The study of cell morphology has contributed to the innovation of clinical techniques and biomedical research. Primary cell culture techniques are well standardized; however, knowledge about morphometric parameters under cell culture conditions is scarce. Variations in morphology can affect cell physiology and responses. The aim of this study was to use morphometric tools to describe the growth and development of skeletal muscle cells under standard cell culture conditions. A photographic database was generated, and morphometric data was obtained for nine cell characteristics (n = 559 cells). Four muscular cell shapes (spherical, irregular outline, triangular and spindle/fusiform) were characterized with wide ranges in variation. The maximum cell length (110-262 µm), width (35-66 µm), area (2,642 - 9,480 µm2), projection lengths (45 - 127 µm), and nucleus diameter (28 ± 11 µm) were obtained by day 23 of culture. A single centrally positioned nucleus was observed in each cell; nucleoli diameter (5 ± 2 µm) and number (1 - 5) varied. In general, cyclic changes in cell sizes were identified during culture, whereas cell length, width, and area increased in spurts. These results suggest that morphometric parameters can be used to monitor skeletal muscle cell development under standard culture conditions.

RESUMEN: A partir de células madre musculares, surgen los mioblastos que se dividen y fusionan entre sí para formar a los miocitos. Estas células ya diferenciadas son precursoras de miocitos que maduran en fibras musculares y posteriormente forman los músculos. La implementación de cultivos celulares de mioblastos ha permitido obtener conocimiento detallado del tejido muscular. Particularmente, algunas de las aportaciones morfológicas fueron el punto de partida de técnicas clínicas, terapias o investigaciones biomédicas. Sin embargo, los estudios morfométricos en condiciones de cultivo celular son escasos. Por lo cual, realizamos seguimientos fotográficos a cultivos desarrollados bajo condiciones estándar, registramos datos para nueve características celulares y aplicamos técnicas morfométricas para analizar estas células (n = 559). Se caracterizaron cuatro formas celulares adoptadas por los mioblastos (esférica, irregular, triangular y huso) y se registraron intervalos amplios de variación en los caracteres. Hacia el día 23 de cultivo se presentaron los valores máximos en la longitud (110-262 µm), el ancho (35-66 µm) y el área celular (2,642-9,480 µm2), así como en el tamaño máximo de las proyecciones celulares (45-127 µm) y el diámetro del núcleo (28±11 µm). El núcleo se observó como único y en posición central; los nucleolos variaron poco en diámetro (5±2 µm), aunque no en número (1 a 5). En términos generales, se identificaron cambios cíclicos en la talla de las células durante los cultivos, esto es, períodos intercalados de incremento y decremento en el largo, ancho y área celular. Debido a que estas características reflejaron los cambios generales sufridos por los mioblastos durante el cultivo, se proponen para monitorear sus etapas de desarrollo en cultivo.

Antibacterial effect of acetic acid during an outbreak of carbapenem-resistant Acinetobacter baumannii in an ICU (II).

INTRODUCTION: Acetic acid (AA) has been commonly used in medicine as an antiseptic agent for the past 6000 years. This study evaluated the antibacterial effect of AA during an outbreak in an intensive care unit (ICU) facility in Baja California Sur, México. METHODOLOGY: Thirty-five environmental samples were collected, subsequently, disinfection with AA (4%) was performed, and two days later the same areas were sampled inside the ICU facility. Carbapenem-resistant A. baumannii (CRAB) was detected with loop-mediated isothermal amplification assay (Garciglia-Mercado et al. companion paper), targeting blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaIMP and blaVIM genes. CRAB isolates before and after disinfection were compared by PFGE. RESULTS: Eighteen (54.5%) and five (14.3%) of thirty-five environmental samples were identified as Acinetobacter baumannii before and after disinfection, respectively, showing a significant decrease of 85.7% (p < 0.05) both by Loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR). Furthermore, the presence of blaOXA-23-like and blaOXA-58-like genes significantly decreased (p < 0.05) both by LAMP and PCR methods. PFGE genotype showed high similarity among CRAB isolates before and after disinfection, suggesting wide clonal dissemination in the ICU facility. CONCLUSIONS: This study demonstrated the novel application of AA with the LAMP assays developed for detecting CRAB. AA promises to be a cheap and efficacious disinfectant alternative to both developed and especially developing countries, preventing the spread of this organism in the environment and to other susceptible patients in health care settings.

In silico Characterization of the Heme Oxygenase 1 From Bottlenose Dolphin (Tursiops truncatus): Evidence of Changes in the Active Site and Purifying Selection.

Cetacea is a clade well-adapted to the aquatic lifestyle, with diverse adaptations and physiological responses, as well as a robust antioxidant defense system. Serious injuries caused by boats and fishing nets are common in bottlenose dolphins (Tursiops truncatus); however, these animals do not show signs of serious infections. Evidence suggests an adaptive response to tissue damage and associated infections in cetaceans. Heme oxygenase (HO) is a cytoprotective protein that participates in the anti-inflammatory response. HO catalyzes the first step in the oxidative degradation of the heme group. Various stimuli, including inflammatory mediators, regulate the inducible HO-1 isoform. This study aims to characterize HO-1 of the bottlenose dolphin in silico and compare its structure to the terrestrial mammal protein. Upstream HO-1 sequence of the bottlenose dolphin was obtained from NCBI and Ensemble databases, and the gene structure was determined using bioinformatics tools. Five exons and four introns were identified, and proximal regulatory elements were detected in the upstream region. The presence of 10 α-helices, three 310 helices, the heme group lodged between the proximal and distal helices, and a histidine-25 in the proximal helix serving as a ligand to the heme group were inferred for T. truncatus. Amino acid sequence alignment suggests HO-1 is a conserved protein. The HO-1 "fingerprint" and histidine-25 appear to be fully conserved among all species analyzed. Evidence of positive selection within an α-helix configuration without changes in protein configuration and evidence of purifying selection were found, indicating evolutionary conservation of the coding sequence structure.

Mercury concentrations in Baja California Sur fish: Dietary exposure assessment.

Total mercury concentrations ([THg]) in muscle were determined in commercial finfish and elasmobranchs from Baja California Sur (BCS), Mexico to evaluate dietary Hg exposure for BCS communities, including the relationship of trophic ecology, length and mass with [THg] that might drive future consumption advice (e.g., recommend limited consumption of large fish for some species). The [THg] ranged from 0.06 to 528.02 µg kg-1 ww in finfish and 17.68-848.26 µg kg-1 ww in elasmobranchs. Relative to the consumption threshold set for predatory fish in Mexico, all species had a concentration below 1000 µg kg-1 ww. As expected, 16 (4.02%) and 75 (18.84%) individual fish were above advisory thresholds of 500 and 200 µg kg-1 ww, respectively. The hazard quotients (HQs) in most species were significantly <1.0, only banded guitarfish showed a significant median HQ > 1.0. Thus, the relative level of risk of high Hg exposure is low for most species.

Development of a LAMP method for detection of carbapenem-resistant Acinetobacter baumannii during a hospital outbreak.

INTRODUCTION: Carbapenem-resistant A. baumannii (CRAB) represents a public health threat increasing worldwide. We assess the suitability of a loop-mediated isothermal amplification (LAMP) method for on-site screening of CRAB in a hospital facility. METHODOLOGY: A set of six primers were designed for recognizing eight distinct sequences on six targets: blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaIMP, and blaVIM. A LAMP method was developed, optimized and evaluated for the identification of CRAB in thirty-three environmental samples from an outbreak in an Intensive Care Unit (ICU) facility. RESULTS: The sensitivity of the LAMP assay for the detection of A. baumannii was ten-fold higher than the PCR assay (1.0 ng.µL-1). The LAMP assays showed a higher detection rate for CRAB samples and robust diagnosis performance in comparison to a conventional PCR, with clinical sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 100% for blaOXA-23-like, blaOXA-51-like and blaVIM. CONCLUSIONS: The developed LAMP assays are powerful tools that can be useful in on-site screening of CRAB causing local outbreaks in clinics and hospitals facilities where costs and equipment restraints are imperative.

Antioxidant response to cadmium exposure in primary skeletal muscle cells isolated from humans and elephant seals.

Cadmium (Cd) occurs naturally; however, its concentration can increase with anthropogenic activities. Excess Cd increases reactive oxygen species (ROS) production and oxidative damage, which can lead to pathological conditions. Marine mammals accumulate Cd in the liver and the kidney; yet, there are no reports of Cd-associated tissue damage in whales, seals or dolphins. Response to Cd exposure (0-5.0 µM CdCl2 for 1-12 h) was analyzed and compared in primary skeletal muscle cells isolated from northern elephant seals (Mirounga angustirostris) and humans (Homo sapiens). Antioxidant enzyme activities (glutathione S-transferase, glutathione reductase, glutathione peroxidase), glutathione concentration, and protein carbonyl levels (an indicator of oxidative damage) were quantified. Glutathione levels were higher in northern elephant seal than in human cells. Protein carbonyl content in cells exposed to Cd was lower and had a smaller variability range in elephant seals than in humans. Generalized linear models (GLIM) identified Cd exposure and antioxidant defenses as significant contributors to protein carbonyl variability in human but not in elephant seal cells. These results suggest that the previously observed differences in circulating and tissue glutathione levels between marine and terrestrial mammals are maintained under cell culture conditions and that northern elephant seal and human muscle cells respond differently to Cd exposure. The results also suggest that the observed differences could potentially be associated with the protective mechanisms that allow northern elephant seals to tolerate extreme conditions that result in increased ROS generation (e.g. diving, sleep apnea, fasting) with no oxidative damage.

Using carbon and nitrogen stable isotope modelling to assess dietary mercury exposure for pregnant women in Baja California Sur, Mexico.

INTRODUCTION: Previous studies of mercury (Hg) in pregnant women in the area of La Paz, Baja California Sur (BCS), Mexico found a proportion of individuals had concentrations of total Hg ([THg]) above some thresholds of concern set by health agencies. The [THg] were associated with fish and seafood consumption as well as other factors; although it was unclear which marine diet items could potentially be contributing to the concentrations observed. METHOD: We examined [THg] and monomethylmercury concentration ([MeHg+]) in the archived hair of 70 pregnant women from BCS as well as in diet items including fish, shellfish, and staple items (rice, beans, corn, and flour). We measured stable isotopes of carbon and nitrogen and employed a Bayesian stable isotope mixing model to investigate the proportion of fish and seafood in the isotopic profiles of archived hair samples. RESULTS: Concentrations of Hg species were low in staple foods and ranged from below detection limit to 5.71 parts per billion (ppb) wet weight. In hair, geometric mean [THg] was 658 ppb and [MeHg+] was 395 ppb, which were lower than previous reports. Percent MeHg+ was positively correlated with higher δ15N values. CONCLUSIONS: The largest carbon contributors to the diet of the study participants were corn and rice, and our analysis of fish contribution to diet varyingly agreed with the self-reported fish consumption. This report highlights the ability to discriminate potential sources of Hg from a diverse diet and the limitations of dietary recall studies.

Micronutrient content and antioxidant enzyme activities in human breast milk.

Breast milk contains micronutrients that function as cofactors of antioxidant enzymes. High concentrations of iron (Fe) and copper (Cu) can increase the production of reactive oxygen species (ROS). This study aimed to assess the relationship between the activity of antioxidant enzymes (superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S-transferase (GST)) and the concentration of the micronutrients Fe, Cu and zinc (Zn) in breast milk. Breast milk samples were collected from 108 mothers (7-10 days postpartum, transitional milk). The samples were grouped into three groups according to the number of pregnancies (one, two and three or more pregnancies), also grouped according to the body mass index (BMI) suggested by the World Health Organization (WHO) in underweight, normal weight, overweight and obese. Breast milk Fe, Cu and Zn concentrations were determined by atomic absorption spectrophotometry and the activity of the antioxidant enzymes was determined by spectrophotometry. An increase in GPx, SOD and GST activities in relation to the number of pregnancies was found (p = 0.05, p = 0.04 and p < 0.01, respectively). An inverse relationship between GST activity and BMI was found (p = 0.05). A positive correlation was observed between Cu and Zn concentrations (r = 0.52, p < 0.05). A negative correlation was found between Cu concentration and catalase activity (r = -0.22, p < 0.05); Fe content was negatively correlated with GPx and GST activities (r = -0.32, r = -0.22, respectively, p < 0.05). The activities of antioxidant enzymes (GPx, SOD and GST) may be affected by the number of pregnancies and contribute to prevent oxidation of nutritional molecules in breast milk.

Oxidative damage to proteins related to metals and antioxidant defenses in breastmilk.

INTRODUCTION: Breast milk contains molecules needed for the development of children; the integrity and function of these molecules is affected by the presence of pro-oxidants. Protein carbonyls are mainly produced as a result of the interaction of metals with reactive oxygen species (ROS), which may initiate a chain reaction that promotes molecular oxidation. OBJECTIVE: This study aimed to determine the association between the concentration of protein carbonyls with the concentration of trace elements (lead [Pb], cadmium [Cd] and selenium [Se]), superoxide radical (O2•-) production, and glutathione (GSH) content, as well with the activity of the main antioxidant enzymes (superoxide dismutase [SOD], catalase [CAT], glutathione peroxidase [GPx], glutathione reductase [GR] and glutathione S-transferase [GST]) in breast milk. METHODS: In this study 108 transitional milk samples (7-10 days) were analyzed. Antioxidant enzyme activities, O2•-production, protein carbonyl and GSH concentrations were analyzed by spectrophotometry. Trace element concentration was quantified by atomic absorption spectrophotometry. Generalized linear modelling was used to assess the relationship between protein carbonyls concentration with oxidative stress indicators and trace elements concentration. RESULTS: Cd and Pb were detected in 21.3 and 55.6% of breast milk samples, respectively. The median concentration of Cd was 0.01 µg L-1 (0.01-3.52 µg L-1) and Pb concentration was 2.61 µg L-1 (0.08-195.20 µg L-1). According to the best-fit model, the main factors contributing to protein carbonyl concentrations were the activity of GPx, GR, and concentration of GSH, Se, Pb and Cd. CONCLUSIONS: According to the generalized linear model, the activity of GPx and GR, could help explain protein oxidation induced by Pb and Cd in breast milk.

Oxidative damage to proteins related to metals and antioxidant defenses in breastmilk / Daño oxidativo a las proteínas relacionado con metales y defensas antioxidantes en la leche materna

Introduction: Breast milk contains molecules needed for the development of children; the integrity and function of these molecules is affected by the presence of pro-oxidants. Protein carbonyls are mainly produced as a result of the interaction of metals with reactive oxygen species (ROS), which may initiate a chain reaction that promotes molecular oxidation. Objective: This study aimed to determine the association between the concentration of protein carbonyls with the concentration of trace elements (lead [Pb], cadmium [Cd] and selenium [Se]), superoxide radical (O2 •-) production, and glutathione (GSH) content, as well with the activity of the main antioxidant enzymes (superoxide dismutase [SOD], catalase [CAT], glutathione peroxidase [GPx], glutathione reductase [GR] and glutathione S-transferase [GST]) in breast milk. Methods: In this study 108 transitional milk samples (7-10 days) were analyzed. Antioxidant enzyme activities, O2 •- production, protein carbonyl and GSH concentrations were analyzed by spectrophotometry. Trace element concentration was quantified by atomic absorption spectrophotometry. Generalized linear modelling was used to assess the relationship between protein carbonyls concentration with oxidative stress indicators and trace elements concentration. Results: Cd and Pb were detected in 21.3 and 55.6% of breast milk samples, respectively. The median concentration of Cd was 0.01 μg L-1 (0.01-3.52 μg L-1) and Pb concentration was 2.61 μg L-1 (0.08-195.20 μg L-1). According to the best-fi t model, the main factors contributing to protein carbonyl concentrations were the activity of GPx, GR, and concentration of GSH, Se, Pb and Cd. Conclusions: According to the generalized linear model, the activity of GPx and GR, could help explain protein oxidation induced by Pb and Cd in breast milk (AU)

Introducción: la leche materna contiene las moléculas necesarias para el desarrollo de los niños; la integridad y función de estas moléculas se afecta por la presencia de prooxidantes. Los carbonilos proteicos se producen principalmente como resultado de la interacción de metales con especies reactivas de oxígeno (ERO), los cuales pueden iniciar una reacción en cadena que promueve la oxidación molecular. Objetivo: este estudio tiene como objetivo determinar la asociación entre la concentración de carbonilos proteicos con la concentración de elementos traza (plomo [Pb], cadmio [Cd] y selenio [Se]), producción de radical superóxido (O2 •-), y contenido de glutatión (GSH), así como con la actividad de las principales enzimas (superóxido dismutasa [SOD], catalasa [CAT], glutatión peroxidasa [GPx], glutatión reductasa [GR] y glutatión S-transferasa [GST]) en leche materna. Métodos: en este estudio se analizaron 108 muestras de leche de transición (7-10 días). La actividad de las enzimas antioxidantes, producción de O2 •-, concentración de carbonilos proteicos y GSH se analizaron por espectrofotometría. La concentración de elementos traza se cuantificó por espectrometría de absorción atómica. Se utilizó un modelo lineal generalizado para evaluar la relación entre la concentración de carbonilos proteicos con los indicadores de estrés oxidativo y las concentraciones de elementos traza. Resultados: Cd y Pb fueron detectados en 21,3 y 55,6% de las muestras de leche materna, respectivamente. La mediana de la concentración de Cd fue 0,01 μg l-1 (0,01-3,52 μg l-1) y para la concentración de Pb fue 2,61 μg l-1 (0,08-195,20 μg l-1). De acuerdo con el modelo de mejor ajuste, los principales factores de afectan la concentración de carbonilos proteicos, son la actividad de GPx y GR, y las concentraciones de GSH, Se, Pb y Cd. Conclusiones: de acuerdo con el modelo lineal generalizado, la actividad de GPx y GR podría ayudar a explicar la oxidación proteica, inducida por Pb y Cd en leche materna (AU)

Plasma Hypoxanthine-Guanine Phosphoribosyl Transferase Activity in Bottlenose Dolphins Contributes to Avoiding Accumulation of Non-recyclable Purines.

Marine mammals are exposed to ischemia/reperfusion and hypoxia/reoxygenation during diving. During oxygen deprivation, adenosine triphosphate (ATP) breakdown implies purine metabolite accumulation, which in humans is associated with pathological conditions. Purine recycling in seals increases in response to prolonged fasting and ischemia. Concentrations of metabolites and activities of key enzymes in purine metabolism were examined in plasma and red blood cells from bottlenose dolphins (Tursiops truncatus) and humans. Hypoxanthine and inosine monophosphate concentrations were higher in plasma from dolphins than humans. Plasma hypoxanthine-guanine phosphoribosyl transferase (HGPRT) activity in dolphins suggests an elevated purine recycling rate, and a mechanism for avoiding accumulation of non-recyclable purines (xanthine and uric acid). Red blood cell concentrations of hypoxanthine, adenosine diphosphate, ATP and guanosine triphosphate were lower in dolphins than in humans; adenosine monophosphate and nicotinamide adenine dinucleotide concentrations were higher in dolphins. HGPRT activity in red blood cells was higher in humans than in dolphins. The lower concentrations of purine catabolism and recycling by-products in plasma from dolphins could be beneficial in providing substrates for recovery of ATP depleted during diving or vigorous swimming. These results suggest that purine salvage in dolphins could be a mechanism for delivering nucleotide precursors to tissues with high ATP and guanosine triphosphate requirements.

[Not Available]. / Antioxidantes, especies reactivas de oxígeno y daño oxidativo asociado a la presencia de plaguicidas organoclorados en la leche materna.

Introducción: los plaguicidas organoclorados (POC) pueden incrementar la producción de especies reactivas de oxígeno (ERO). Tales efectos pueden ser contrarrestados por el sistema antioxidante, el cual se encuentra también en la leche materna. Objetivo: evaluar los indicadores de estrés oxidativo en leche materna asociados a la presencia de POC y su relación con la ingesta de alimentos marinos. Métodos: la leche materna fue colectada de 108 mujeres lactantes. Los indicadores de estrés oxidativo (actividad enzimática antioxidante, concentración de glutatión, producción de radical superóxido [O 2•- ], concentración de peroxidación de lípidos y carbonilos proteicos) se analizaron por espectrofotometría. Las concentraciones de POC se midieron por cromatografía de gases. Resultados y discusión: la producción de O 2•- no presentó relación significativa con las concentraciones de POC. Se encontraron correlaciones significativas entre las concentraciones de POC y la actividad de las enzimas antioxidantes (actividad de glutatión reductasa [GR] y concentración de aldrín [r = - 0,5], actividad de superóxido dismutasa [SOD] y concentración de α-HCH [r = 0,45]). El daño oxidativo mostró baja correlación con el contenido de POC (r < 0,30, p < 0,05). Es posible que los niveles de POC no sean suficientes para incrementar la producción de O 2•- , ya sea que el incremento en la producción de ERO se deba a especies reactivas diferentes a O 2•- o debido a que la capacidad antioxidante es suficiente para evitar el daño oxidativo en leche materna. Conclusión: los resultados de este estudio sugieren que la dieta marina no es un factor determinante en el nivel de contaminación por POC, ni en el daño oxidativo presente en leche materna.

Antioxidants, reactive oxygen species and oxidative damage associated to the presence of organochlorine pesticides in breast milk / Antioxidantes, especies reactivas de oxígeno y daño oxidativo asociado a la presencia de plaguicidas organoclorados en la leche materna

Introduction: Organochlorine pesticides (OCPs) can increase the production of reactive oxygen species (ROS), the effects of which can be countered by the antioxidant system, also present in breast milk. Objective: To evaluate the indicators of oxidative stress in breast milk associated to the presence of OCPs, and their relationship with seafood intake. Methods: Breast milk was collected from 108 lactating women. Indicators of oxidative stress (antioxidant enzyme activity, glutathione concentration, superoxide radical [O2 •-] production, lipid peroxidation and carbonylated protein concentration) were analyzed by spectrophotometry. OCPs concentrations were measured by gas chromatography. Results and discussion: O2 •- production had no significant relationship with OCPs concentrations. Significant correlations between OCPs concentrations and antioxidant enzyme activities (glutathione reductase [GR] activity and aldrin concentration [r = - 0.5], superoxide dismutase [SOD] activity and α-HCH concentration [r = 0.45]) were found. Oxidative damage had a low correlation with OCPs content (r < 0.30, p < 0.05). It is possible that OCP’s levels are not sufficient to increase O2 •- production, that production of ROS other than O2 •- is increased, or that the antioxidant capacity is sufficient to avoid oxidative damage in breast milk. Conclusion: Results from this study suggest that marine diet is not a determinant factor in the level of contamination by OCP’s or in the oxidative damage in breast milk (AU)

Introducción: los plaguicidas organoclorados (POC) pueden incrementar la producción de especies reactivas de oxígeno (ERO). Tales efectos pueden ser contrarrestados por el sistema antioxidante, el cual se encuentra también en la leche materna. Objetivo: evaluar los indicadores de estrés oxidativo en leche materna asociados a la presencia de POC y su relación con la ingesta de alimentos marinos. Métodos: la leche materna fue colectada de 108 mujeres lactantes. Los indicadores de estrés oxidativo (actividad enzimática antioxidante, concentración de glutatión, producción de radical superóxido [O2 •-], concentración de peroxidación de lípidos y carbonilos proteicos) se analizaron por espectrofotometría. Las concentraciones de POC se midieron por cromatografía de gases. Resultados y discusión: la producción de O2 •- no presentó relación significativa con las concentraciones de POC. Se encontraron correlaciones significativas entre las concentraciones de POC y la actividad de las enzimas antioxidantes (actividad de glutatión reductasa [GR] y concentración de aldrín [r = - 0,5], actividad de superóxido dismutasa [SOD] y concentración de α-HCH [r = 0,45]). El daño oxidativo mostró baja correlación con el contenido de POC (r < 0,30, p < 0,05). Es posible que los niveles de POC no sean suficientes para incrementar la producción de O2 •-, ya sea que el incremento en la producción de ERO se deba a especies reactivas diferentes a O2 •- o debido a que la capacidad antioxidante es suficiente para evitar el daño oxidativo en leche materna. Conclusión: los resultados de este estudio sugieren que la dieta marina no es un factor determinante en el nivel de contaminación por POC, ni en el daño oxidativo presente en leche materna (AU)

The Neuroprotective Effect of Erythropoietin in Rat Hippocampus in an Endotoxic Shock Model.

BACKGROUND: Sepsis is characterized by an early systemic inflammation in response to infection. In the brain, inflammation is associated with expression of pro-inflammatory cytokines (e.g. tumor necrosis factor-α, interleukin-1ß and interleukin-6, among others) that may induce an overproduction of reactive oxygen and nitrogen species. The constitutive expression of cytokines in the brain is low, but may be induced by various stimuli, including lipopolysaccharide, which causes neuronal damage. Erythropoietin, among other effects, acts as a multifunctional neurotrophic factor implicated in neurogenesis, angiogenesis, vascular permeability, and immune regulation in the central nervous system. In an experimental model of endotoxic shock, we studied the neuroprotective capacity of erythropoietin in the rat hippocampus and compared with melatonin, a neurohormone with an important antioxidant and immunomodulatory effect. METHODS: In 21-day-old male Wistar rats divided into eight groups, we administered by intraperitoneal injection lipopolysaccharide, erythropoietin, melatonin, or combinations thereof. The hippocampus was dissected and morphological (histological analysis) and biochemical (cytokine levels) studies were conducted. RESULTS: The number of dead neuronal cells in histological sections in groups treated with lipopolysaccharide was higher compared to the erythropoietin group. There was a greater decrease (70%) in interleukin-1ß concentrations in rats with endotoxic shock that received erythropoietin compared to the lipopolysaccharide group. CONCLUSIONS: The neuronal cell loss caused by endotoxic shock and interleukin-1ß levels were reduced by the administration of the hematopoietic cytokine erythropoietin in this experimental model.

Purine nucleoside phosphorylase and the enzymatic antioxidant defense system in breast milk from women with different levels of arsenic exposure.

Purine nucleoside phosphorylase (PNP) is an ubiquitous enzyme which plays an important role in arsenic (As) detoxification. As is a toxic metalloid present in air, soil and water; is abundant in the environment and is readily transferred along the trophic chain, being found even in human breast milk. Milk is the main nutrient source for the growth and development of neonates. Information on breast milk synthesis and its potential defense mechanism against As toxicity is scarce. In this study, PNP and antioxidant enzymes activities, as well as glutathione (GSH) and total arsenic (TAs) concentrations, were quantified in breast milk samples. PNP, superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR) activities and GSH concentration were determined spectrophotometrically; TAs concentration ([TAs]) was measured by atomic absorption spectrometry. Data suggest an increase in PNP activity (median = 0.034 U mg protein-1) in the presence of TAs (median = 1.16 g L(-1)). To explain the possible association of PNP activity in breast milk with the activity of the antioxidant enzymes as well as with GSH and TAs concentrations, generalized linear models were built. In the adjusted model, GPx and GR activities showed a statistically significant (p<0.01) association with PNP activity. These results may suggest that PNP activity increases in the presence of TAs as part of the detoxification mechanism in breast milk.

Purina nucleósido fosforilasa (PNP) es una enzima ubicua que desempeña un papel importante en la desintoxicación del arsénico (As). As es un metaloide tóxico presente en el aire, el suelo y el agua; es abundante en el medio ambiente y se transfiere fácilmente a lo largo de la cadena trófica, encontrándose incluso en la leche materna humana. Información sobre la síntesis de la leche materna y su potencial mecanismo de defensa contra tóxicos es escasa. En este estudio, se cuantificó la actividad de PNP y de las enzimas antioxidantes así como la concentración de glutatión (GSH) y de arsénico total ([TAs]) en muestras de leche materna. La actividad de PNP, superóxido dismutasa (SOD), catalasa (CAT), glutatión S-transferasa (GST), glutatión peroxidasa (GPx), glutatión reductasa (GR) y la concentración de GSH se determinaron por espectrofotometría; la [TAs] se midió por espectrometría de absorción atómica. Los datos sugieren un incremento en la actividad de PNP (mediana= 0.034 U mg proteína-1) con la presencia de TAs (mediana= 1.16 g L-1). Para explicar la posible asociación de la actividad de las enzimas antioxidantes y la concentración de GSH, así como [TAs], con la actividad de PNP en la leche materna, se construyeron modelos lineales generalizados. En el modelo ajustado, la actividad de GPx y GR presentó una asociación estadística (p.