Evaluating the Incorporation of Myrtus communis L. Leaves Infusion in Alginate-Based Films and Spheres to Enhance the Oxidative Stability of Oil-in-Water Emulsions.

Myrtus communis L. is a species of the Myrtaceae family that is found in the Mediterranean region, and it is traditionally recognized for its importance and different uses. The objective of this study was to determine the effect of M. communis L. leaf extract (MCLE), which was incorporated directly into alginate spheres and films, on preserving oil-in-water emulsions from oxidation. For this purpose, the solvent extraction (with ethanol at 40, 60, and 80%) of the antioxidant compounds was optimized (total phenolic compounds (TPCs) and total flavonoid content (TFC)) along with the scavenging activity. The best condition for the extraction corresponded with 60% ethanol (MCLE60), with a TPC of ~66.06 g GAE/L and a TFC of ~18.91 g QE/L, which was selected for use in the following assays. MCLE60 showed a considerable radical scavenging activity (24.85 mmol TE/L in FRAP, 28.75 mmol TE/L in DPPH, 30.61 mmol TE/L in ABTS, and 14.94 mmol TE/L in ORAC), which was probably due to its content in the phenolic compounds arbutin (122.08 mg/L), epicatechin (73.89 mg/L), sinapic acid (51.85 mg/L), and gallic acid (36.72 mg/L). The oil-in-water emulsions with the MCLE60 spheres showed the best oxidative stability (TBARS ~2.64 mg MDA/kg of emulsion, PV ~35.7 meq hydroperoxides/kg of emulsion) in comparison to the control. The film was also able to protect the emulsion from oxidation for more than a week at 30 °C (TBARS ~1.9 mg MDA/kg of emulsion). The alginate films with MCLE60 presented an important release of phenolic compounds in water and acetic food simulants, while in both ethanol simulants, the release of TPC remained more stable over time. Thus, this study highlights the potential uses of MCLE as a natural ingredient for emulsion oxidative preservation and the production of alginate delivery systems (spheres and films).

Piezo1 activates noncanonical EGFR endocytosis and signaling.

EGFR-ERK signaling controls cell cycle progression during development, homeostasis, and disease. While EGF ligand and mechanical inputs can activate EGFR-ERK signaling, the molecules linking mechanical force to this axis have remained mysterious. We previously found that stretch promotes mitosis via the stretch-activated ion channel Piezo1 and ERK signaling. Here, we show that Piezo1 provides the missing link between mechanical signals and EGFR-ERK activation. While both EGF- and Piezo1-dependent activation trigger clathrin-mediated EGFR endocytosis and ERK activation, EGF relies on canonical tyrosine autophosphorylation, whereas Piezo1 involves Src-p38 kinase-dependent serine phosphorylation. In addition, unlike EGF, ex vivo lung slices treated with Piezo1 agonist promoted cell cycle re-entry via nuclear ERK, AP-1 (FOS and JUN), and YAP accumulation, typical of regenerative and malignant signaling. Our results suggest that mechanical activation via Piezo1, Src, and p38 may be more relevant to controlling repair, regeneration, and cancer growth than tyrosine kinase signaling via canonical EGF signaling, suggesting an alternative therapeutic approach.

Study of mitochondrial function in thawed bull spermatozoa using selective electron transfer chain inhibitors.

Bull spermatozoa depend equally on glycolysis and oxidative phosphorylation for the maintenance of the energy necessary for their proper functioning. The aim of the present work was to delineate the mitochondrial activity of bull spermatozoa after incubation with specific inhibitors of the different mitochondrial complexes and evaluate their ROS production. Thawed bull sperm cells (30 × 106 mL-1 in Tyrode's extender) were incubated 1 and 3h at 37 °C with rotenone 5 µM (ROT), complex I inhibitor; dimethyl-malonate 10 mM (DMM), complex II inhibitor; carbonyl cyanide m-chlorophenyl hydrazine 5 µM (CCCP), uncoupling agent; antimycin A 1 µg/mL (ANTI), complex III inhibitor; oligomycin 5 µM (OLIGO), ATP synthase inhibitor, and 0.5% DMSO, vehicle (CTR). Sperm motility and kinematics were assessed by Hamilton Thorn IVOS 12.0. Mitochondrial membrane potential, mitochondrial O2•- production and H2O2 intracellular content were evaluated by BD FACSCalibur flow cytometer, and sperm viability (SYBR-14/PI) and mitochondrial activity (JC-1/SYBR-14/PI) were assessed by epifluorescence microscopy. A multivariate analysis was performed on the results. In addition, sperm kinematic features, registered for each motile spermatozoon, were studied by cluster analysis. The incubation during 1 or 3 h in presence of the inhibitors of mitochondrial functionality only had a minor effect on motility parameters, decreasing the proportion of the SP1 (fast progressive) subpopulation after 3 h of incubation with ROT, ANTI or OLIGO. The percentage of live spermatozoa with active mitochondria was reduced under the effect of ANTI and CCCP both at 1 and 3 h. In conclusion, mitochondrial function is somehow impaired in frozen thawed bull sperm as not all live cells showed active mitochondria. These results support the findings that bull spermatozoa can alternatively rely on oxidative phosphorylation or glycolysis for energy obtainment and that their mitochondria are less affected by ETC inhibitors.

Sustainable Food Support during an Ultra-Endurance and Mindfulness Event: A Case Study in Spain.

The present industrial food-production system is not suitably ecological for the environment. Mindful nutrition in sport is a relevant emergent sub-discipline that could help reduce environmental degradation. This case study describes a sustainable support diet during an ultra-endurance running (UR) event called the "Indoor Everest Challenge". This UR challenge involved attaining the altitude of Mount Everest (8849 m) in a simulated way, in less than 24 h, without using ultra-processed food and without wasting plastics. During this challenge, a male athlete (34 years, weight: 78 kg, and height: 173 cm) wore a SenseWear Armband® (BodyMedia Inc., Pittsburg, PA, USA) accelerometer on his right arm to estimate energy expenditure. To supply his nutritional requirements, the athlete consumed only specially prepared homemade and organic food. All consumption was weighed and recorded in real-time; we determined nutrients using two databases: a food composition software, Dial Alce Ingenieria® (Madrid, Spain), to measure energy and macro- and micro-nutrients, and Phenol Explorer Database® (INRA Institut National de Recherche pour l'Alimentation, Paris, France) precisely to determine polyphenolic content. Most energy intake (up to 96%) came from plant foods. We found that subject consumed 15.8 g/kg-1/d-1 or 1242 g of carbohydrates (CHO), (2.4 g/kg-1/d-1) or 190 g of proteins (P), and 10,692 mL of fluid. The total energy intake (7580 kcal) showed a distribution of 65% CHO, 10% P, and 25% lipids (L). Furthermore, this sustainable diet lead to a high antioxidant intake, specifically vitamin C (1079 mg), vitamin E (57 mg), and total polyphenols (1910 mg). This sustainable approach was suitable for meeting energy, CHO, and P recommendations for UR. Physical and mental training (mindfulness) were integrated from the specific preliminary phase to the day of the challenge. The athlete completed this challenge in 18 h with a low environmental impact. This sports event had an educational component, as it awakened curiosity towards food sustainability.

Activation of AMP-activated protein kinase during sepsis/inflammation improves survival by preserving cellular metabolic fitness.

The purpose was to determine the role of AMPK activation in the renal metabolic response to sepsis, the development of sepsis-induced acute kidney injury (AKI) and on survival. In a prospective experimental study, 167 10- to 12-week-old C57BL/6 mice underwent cecal ligation and puncture (CLP) and human proximal tubule epithelial cells (TEC; HK2) were exposed to inflammatory mix (IM), a combination of lipopolysaccharide (LPS) and high mobility group box 1 (HMGB1). Renal/TEC metabolic fitness was assessed by monitoring the expression of drivers of oxidative phosphorylation (OXPHOS), the rates of utilization of OXPHOS/glycolysis in response to metabolic stress, and mitochondrial function by measuring O2 consumption rates (OCR) and the membrane potential (Δψm ). Sepsis/IM resulted in AKI, increased mortality, and in renal AMPK activation 6-24 hours after CLP/IM. Pharmacologic activation of AMPK with 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) or metformin during sepsis improved the survival, while AMPK inhibition with Compound C increased mortality, impaired mitochondrial respiration, decreased OCR, and disrupted TEC metabolic fitness. AMPK-driven protection was associated with increased Sirt 3 expression and restoration of metabolic fitness. Renal AMPK activation in response to sepsis/IM is an adaptive mechanism that protects TEC, organs, and the host by preserving mitochondrial function and metabolic fitness likely through Sirt3 signaling.

Oligomerization of equilibrative nucleoside transporters: a novel regulatory and functional mechanism involving PKC and PP1.

Equilibrative nucleoside transporters (ENTs) translocate nucleosides and nucleobases across plasma membranes, as well as a variety of anti-cancer, -viral, and -parasite nucleoside analogs. They are also key members of the purinome complex and regulate the protective and anti-inflammatory effects of adenosine. Despite their important role, little is known about the mechanisms involved in their regulation. We conducted membrane yeast 2-hybrid and coimmunoprecipitation studies and identified, for the first time to our knowledge, the existence of protein-protein interactions between human ENT1 and ENT2 (hENT1 and hENT2) proteins in human cells and the formation of hetero- and homo-oligomers at the plasma membrane and the submembrane region. The use of NanoLuc Binary Technology allowed us to analyze changes in the oligomeric status of hENT1 and hENT2 and how they rapidly modify the uptake profile for nucleosides and nucleobases and allow cells to respond promptly to external signals or changes in the extracellular environment. These changes in hENTs oligomerization are triggered by PKC activation and subsequent action of protein phosphatase 1.-Grañe-Boladeras, N., Williams, D., Tarmakova, Z., Stevanovic, K., Villani, L. A., Mehrabi, P., Siu, K. W. M., Pastor-Anglada, M., Coe, I. R. Oligomerization of equilibrative nucleoside transporters: a novel regulatory and functional mechanism involving PKC and PP1.

Simvastatin and metformin inhibit cell growth in hepatitis C virus infected cells via mTOR increasing PTEN and autophagy.

Hepatitis C virus (HCV) infection has been related to increased risk of development of hepatocellular carcinoma (HCC) while metformin (M) and statins treatment seemed to protect against HCC development. In this work, we aim to identify the mechanisms by which metformin and simvastatin (S) could protect from liver cancer. Huh7.5 cells were infected with HCV particles and treated with M+S. Human primary hepatocytes were treated with M+S. Treatment with both drugs inhibited Huh7.5 cell growth and HCV infection. In non-infected cells S increased translational controlled tumor protein (TCTP) and phosphatase and tensin homolog (PTEN) proteins while M inhibited mammalian target of rapamycin (mTOR) and TCTP. Simvastatin and metformin co-administered down-regulated mTOR and TCTP, while PTEN was increased. In cells infected by HCV, mTOR, TCTP, p62 and light chain 3B II (LC3BII) were increased and PTEN was decreased. S+M treatment increased PTEN, p62 and LC3BII in Huh7.5 cells. In human primary hepatocytes, metformin treatment inhibited mTOR and PTEN, but up-regulated p62, LC3BII and Caspase 3. In conclusion, simvastatin and metformin inhibited cell growth and HCV infection in vitro. In human hepatocytes, metformin increased cell-death markers. These findings suggest that M+S treatment could be useful in therapeutic prevention of HCV-related hepatocellular carcinoma.

Greenhouse gas emissions and soil organic matter dynamics in woody crop orchards with different irrigation regimes.

Water scarcity in arid, semiarid and dry regions is a limiting factor for the development of sustainable agriculture. As a consequence, the adoption of new strategies such as regulated deficit irrigation (RDI) to reduce water and energy consumption will be essential. Decreases in irrigation water content may also have positive effects on soil C cycle. Thus, an experiment was setup in three woody crop orchards during two years, with the objective of assessing if RDI can reduce soil CO2 and N2O emissions, modify soil inorganic C and organic C quality and stability and affect soil aggregation. Soil CO2 and N2O emissions were measured every two weeks while soil samplings were carried out every three months. Results indicated that decreases in soil moisture by RDI implementation were related to significant decreases in CO2 emissions in all crops. RDI contributed to an average decrease, compared with full irrigation, of 1088-1664 g CO2 m-2 in the experimental period. Furthermore, CO2 emission was negatively correlated with inorganic C, suggesting the protective effect of soil carbonates towards organic matter. RDI also contributed to significantly decrease soil N2O emissions. However, N2O emission patterns did not directly follow soil moisture patterns and were constant in the experimental period. RDI contributed to an average decrease, compared with full irrigation, of 90-409 mg N2O m-2. No physicochemical property was significantly affected by irrigation regime. Although microbial biomass was not significantly affected by RDI, ß-glucosidase activity was significantly higher under full irrigation during the warm seasons, with significant positive correlation with CO2 emissions. This seems to suggest that a significant fraction of CO2 emitted from soil derives from organic matter degradation, which is limited with low water content. So, RDI could contribute to promote soil C sequestration by reduced greenhouse gas emissions, with no negative effects on soil structure at short-term.

An Allosteric Inhibitor Scaffold Targeting the PIF-Pocket of Atypical Protein Kinase C Isoforms.

There is a current and pressing need for improved cancer therapies. The use of small molecule kinase inhibitors and their application in combinatorial regimens represent an approach to personalized targeted cancer therapy. A number of AGC kinases, including atypical Protein Kinase C enzymes (PKCs), are validated drug targets for cancer treatment. Most drug development programs for protein kinases focus on the development of drugs that bind at the ATP-binding site. Alternatively, allosteric drugs have great potential for the development of future innovative drugs. However, the rational development of allosteric drugs poses important challenges because the compounds not only must bind to a given site but also must stabilize forms of the protein with a desired effect at a distant site. Here we describe the development of a new class of compounds targeting a regulatory site (PIF-pocket) present in the kinase domain and provide biochemical and crystallographic data showing that these compounds allosterically inhibit the activity of atypical PKCs. PS432, a representative compound, decreased the rate of proliferation of non-small cell lung cancer cells more potently than aurothiomalate, an atypical PKCι inhibitor currently under evaluation in clinical trials, and significantly reduced tumor growth without side effects in a mouse xenograft model. The druglike chemical class provides ample possibilities for the synthesis of derivative compounds, with the potential to allosterically modulate the activity of atypical PKCs and other kinases.

Clinically used selective estrogen receptor modulators affect different steps of macrophage-specific reverse cholesterol transport.

Selective estrogen receptor modulators (SERMs) are widely prescribed drugs that alter cellular and whole-body cholesterol homeostasis. Here we evaluate the effect of SERMs on the macrophage-specific reverse cholesterol transport (M-RCT) pathway, which is mediated by HDL. Treatment of human and mouse macrophages with tamoxifen, raloxifene or toremifene induced the accumulation of cytoplasmic vesicles of acetyl-LDL-derived free cholesterol. The SERMs impaired cholesterol efflux to apolipoprotein A-I and HDL, and lowered ABCA1 and ABCG1 expression. These effects were not altered by the antiestrogen ICI 182,780 nor were they reproduced by 17ß-estradiol. The treatment of mice with tamoxifen or raloxifene accelerated HDL-cholesteryl ester catabolism, thereby reducing HDL-cholesterol concentrations in serum. When [(3)H]cholesterol-loaded macrophages were injected into mice intraperitoneally, tamoxifen, but not raloxifene, decreased the [(3)H]cholesterol levels in serum, liver and feces. Both SERMs downregulated liver ABCG5 and ABCG8 protein expression, but tamoxifen reduced the capacity of HDL and plasma to promote macrophage cholesterol efflux to a greater extent than raloxifene. We conclude that SERMs interfere with intracellular cholesterol trafficking and efflux from macrophages. Tamoxifen, but not raloxifene, impair M-RCT in vivo. This effect is primarily attributable to the tamoxifen-mediated reduction of the capacity of HDL to promote cholesterol mobilization from macrophages.

Oxidized low-density lipoprotein antibodies/high-density lipoprotein cholesterol ratio is linked to advanced non-alcoholic fatty liver disease lean patients.

BACKGROUND AND AIM: A small but significant proportion of patients with normal body mass index show non-alcoholic fatty liver disease (NAFLD). Oxidized low-density lipoprotein (LDL) is a powerful immunogenic molecule, which causes oxidative stress and produces antibodies (oxLDL-ab). We aimed to analyze the role of oxLDL-ab on histological features in lean-NAFLD patients. METHODS: Seventy-two biopsy-proven NAFLD patients were included. Lean patients showed body index mass of <30 kg/m(2) . Liver biopsies were assessed by one pathologist blinded to clinical data. Histological features were non-alcoholic steatohepatitis (NASH), steatosis, hepatocellular ballooning, and liver fibrosis. Metabolic and hepatic profiles were analyzed, and lipid-lowering medication was recorded. OxLDL-ab levels were measured by ELISA. OxLDL-ab-based lipid indexes analyzed: oxLDL-ab/total cholesterol ratio; oxLDL-ab/LDL-c ratio; oxLDL-ab/high-density lipoprotein cholesterol (HDL-c) ratio; and oxLDL-ab/oxLDL ratio. RESULTS: Lean-NAFLD patients presented 26.5% (9/34) of NASH. OxLDL-ab/HDL-c ratio (r = 0.570; n = 34; P = 0.001) correlated with NAS score and was the only variable associated with NASH in the multivariate analysis [odds ratio, OR, 1.10 (95% confidence interval, CI: 1.01-1.21); P = 0.039]. Severe steatosis was present in 41.2% (14/34) of lean-NAFLD patients. OxLDL-ab/HDL-c ratio was higher in patients with grade-III steatosis (54.9 (37.3-124.6)) than those with grade II (37.1 (20.2-71.1)) and grade I (17.7 (13.1-22.8)) (P = 0.018). Hepatocellular ballooning was present in 20.6% (7/34) of lean-NAFLD patients, and OxLDL-ab/HDL-c ratio (OR 1.03 [95% CI: 1.01-1.05]; P = 0.050) was independently associated with histological features. OxLDL-ab/HDL-c ratio was higher in patients with advanced fibrosis (39.8 (22.9-121.6) vs 17.7 (13.9-30.9); P = 0.025), increasing gradually with the fibrosis stage (P = 0.042) and remained in the final multivariate model [OR 1.05 (95% CI: 1.00-1.11); P = 0.05]. However, in obese-NAFLD patients, oxLDL/HDL-c ratio was not associated with histological features. CONCLUSIONS: Oxidized low-density lipoprotein antibodies/high-density lipoprotein cholesterol ratio could represent an interesting biomarker associated with NASH, hepatocellular ballooning, and liver fibrosis, in lean patients. OxLDL-ab/HDL-c could play an important role for distinguishing patients with and without NAFLD complications.

Shelf-life of fresh blueberries coated with quinoa protein/chitosan/sunflower oil edible film.

BACKGROUND: The aim of this study was to evaluate quinoa protein (Q), chitosan (CH) and sunflower oil (SO) as edible film material as well as the influence of this coating in extending the shelf-life of fresh blueberries stored at 4 °C and 75% relative humidity. These conditions were used to simulate the storage conditions in supermarkets and represent adverse conditions for testing the effects of the coating. The mechanical, barrier, and structural properties of the film were measured. The effectiveness of the coating in fresh blueberries (CB) was evaluated by changes in weight loss, firmness, color, molds and yeast count, pH, titratable acidity, and soluble solids content. RESULTS: The tensile strength and elongation at break of the edible film were 0.45 ± 0.29 MPa and 117.2% ± 7%, respectively. The water vapor permeability was 3.3 × 10(-12) ± 4.0 × 10(-13) g s(-1) m(-1) Pa(-1). In all of the color parameters CB presented significant differences. CB had slight delayed fruit ripening as evidenced by higher titratable acidity (0.3-0.5 g citric acid 100 g(-1)) and lower pH (3.4-3.6) than control during storage; however, it showed reduced firmness (up to 38%). CONCLUSION: The use of Q/CH/SO as a coating in fresh blueberries was able to control the growth of molds and yeasts during 32 days of storage, whereas the control showed an increasing of molds and yeast, between 1.8 and 3.1 log cycles (between 20 and 35 days).

Cystatin C-based CKD-EPI equations and N-terminal pro-B-type natriuretic peptide for predicting outcomes in acutely decompensated heart failure.

BACKGROUND: In patients with acute decompensated heart failure (ADHF), both natriuretic peptides and renal impairment predict adverse outcomes. Our aim was to evaluate the complementary prognosis role of N-terminal pro-B-type natriuretic peptide (NT-proBNP) and the newly developed Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations based on cystatin C (CysC) for glomerular filtration rate (GFR) estimation in ADHF patients. HYPOTHESIS: Renal impairment assessed by CysC-based CKD-EPI equations and natriuretic peptides have complementary prognostic value in ADHF patients. METHODS: The study included 613 consecutive patients presenting with ADHF. At admission, plasma levels of NT-proBNP and CysC were determined. The GFR was estimated using CysC-based CKD-EPI equations. The primary endpoint was death from any cause and heart failure readmission. RESULTS: During the median follow-up of 365 days (interquartile range, 227-441 days), 323 patients (0.65 %patient-year) died or were readmitted for heart failure. After multivariate adjustment, estimated GFR <60 mL/min/1.73 m(2) and NT-proBNP >3251 pg/mL were independent predictors of adverse outcomes (P < 0.01). The combination of GFR <60 mL/min/1.73 m(2) and NT-proBNP >3251 pg/mL was associated with the highest risk of adverse outcomes. Furthermore, reclassification analyses demonstrated that use of both NT-proBNP and CysC-based CKD-EPI equations resulted in improving the accuracy for adverse outcomes prediction. CONCLUSIONS: In patients with ADHF, the combination of NT-proBNP with estimated GFR using CysC-based CKD-EPI equations better predicts outcomes than either parameter alone and adds valuable complementary prognosis information to other established risk factors.

Cardiometabolic Profile Related to Body Adiposity Identifies Patients Eligible for Bariatric Surgery More Accurately than BMI.

BACKGROUND: Eligibility criteria for bariatric surgery (BS) are based on BMI and the presence of major comorbidities. Our aim was to analyze the usefulness of body adiposity determination in establishing the indication for BS. METHODS: In order to analyze the cardiometabolic risk according to eligibility criteria for BS, four groups were studied. Morbidly obese patients with BMI ≥ 40 kg/m(2) (n = 360), and obese subjects with BMI ≥ 35 kg/m(2) and at least one comorbidity (n = 431), represented two groups of patients meeting original NIH criteria for BS. A third group included two cohorts of patients with a high body fat (BF)% that do not meet the original NIH eligibility criteria for BS: patients with either a BMI <35 kg/m(2) or a BMI ≥ 35 kg/m(2) without comorbidities (n = 266, NEHF). Lean subjects by BMI were the reference group (n = 140). BMI, BF% and markers of insulin sensitivity, lipid profile, and cardiovascular risk were measured. RESULTS: Individuals from the NEHF group exhibited increased HbA1c (P < 0.05) and decreased insulin sensitivity evidenced by a significant reduction in QUICKI (P < 0.001). Triglyceride concentrations were similarly increased (P < 0.05) in the three groups of obese patients. Uric acid concentrations were significantly elevated (P < 0.01) to a similar extent in the obese groups. Levels of the inflammatory marker CRP and hepatic enzymes were significantly increased in the three obese groups. CONCLUSION: The present study provides evidence for the existence of an adverse cardiometabolic profile in subjects currently considered to be outside traditional NIH guidelines but exhibiting a highly increased adiposity. It is concluded that body composition analysis yields valuable information to be incorporated into indication criteria for BS and that adiposity may be an independent indicator for BS.

Delimitación de fronteras como estrategia de control social: el caso de la violencia homicida en Medellín, Colombia / Marking bourdaries as a strategy of social control: the case of homicidal violence in Medellin, Colombia

En el marco de un estudio realizado entre 2003 y 2011 para comprender escenarios de violencia homicida a partir de la percepción del hecho violento y su contexto, se reflexiona sobre el sentido de las "fronteras invisibles" en barrios de Medellín (Colombia). Desde un enfoque cualitativo que combina revisión documental y entrevistas, se analiza la experiencia vivida por ocho participantes. Entre los principales resultados se destaca que el control barrial es ejercido por distintos actores; que las fronteras no son visibles para el común de las personas, sino que son demarcaciones en las que son reclutadas y controladas, y que consolidan estrategias para recaudar recursos económicos de forma ilegal y regular las actividades culturales y sociales de los habitantes, lo cual repercute en la dinámica y los imaginarios sociales. De este modo, se controlan los territorios, las amistades y los afectos de víctimas jóvenes -que no se vinculan a grupos ilegales y/o no tienen "información"- y de adultos mayores indefensos.

As part of a research study undertaken in the period 2003-2011 to understand situations of homicidal violence based in perceptions regarding the act of violence and the surrounding context, we reflect on the meaning of "invisible bourdaries" in the neighborhoods of Medellin (Colombia). Using a qualitative approach that combines documentary sources and interviews, the experiences of 8 participants are analyzed. In the primary results we can see how control over neighborhoods is exercised by different actors through bourdaries not visible to ordinary people. Nevertheless, around these lines people are recruited and controlled and strategies to illegally generate economic resources and to regulate the cultural and social activities of inhabitants are consolidated, thus affecting the social dynamics and imaginary of the neighborhood. In this way, the territories, friendships, and affects of young victims - who are not linked to illegal groups and/or do not have "information" - and of defenseless older adults are controlled.

Essential oil from Chenopodium ambrosioides and main components: activity against Leishmania, their mitochondria and other microorganisms.

Chenopodium ambrosioides is an aromatic herb used by native people to treat parasitic diseases. The aim of this work is to compare the in vitro anti-leishmanial activity of the essential oil (EO) from C. ambrosioides and its major components (ascaridole, carvacrol and caryophyllene oxide) and study their mechanism of action and activity against a panel of microorganism. Antileishmanial activity and cytotoxicity of the EO and major components was study. In addition, experiments to elucidate the mechanism of action were perform and activities against other microorganisms (bacteria, fungi and protozoa) were evaluate. All products were active against promastigote and amastigote forms of Leishmania. Ascaridole exhibited the better antileishmanial activity and the EO the highest selectivity index. The exploration of the mechanism suggests that the products cause a breakdown of mitochondrial membrane potential and a modification of redox indexes. Only EO showed antiprotozoal effect against Plasmodium falciparum and Trypanosoma brucei; while no activity against bacteria and fungi was observed. Our results demonstrate the potentialities of EO in cellular and molecular system, which could be consider in future studies to develop new antileishmanial drugs with a wide anti-parasitic spectrum.

Effect of several antioxidants on thawed ram spermatozoa submitted to 37°C up to four hours.

Thawed ram spermatozoa were incubated at 37°C in the presence of dehydroascorbic acid (DHA), TEMPOL (TPL), N-acetyl-cysteine (NAC) and rutin (RUT), at 0.1 and 1 mm, in order to test their effects on sperm physiology. Cryopreserved spermatozoa from four rams were thawed, pooled, washed and incubated in TALP-Hepes with 1 mm or 0.1 mm of each antioxidant, performing a replicate with induced oxidative stress (Fe(2+) /ascorbate). Motility (CASA), viability and mitochondrial membrane potential (flow cytometry) were analysed at 2 and 4 h. Lipoperoxidation (MDA production), intracellular reactive oxygen species (ROS) and DNA status (TUNEL) were analysed at 4 h. Antioxidants, except DHA 0.1 mm, decreased motility and kinematic parameters, but had little effect on viability or mitochondrial activity. Except 1 mm DHA, the antioxidants reduced ROS at 4 h. Moreover, NAC 1 mm, rutin and TEMPOL reduced ROS and DNA damage in the presence of oxidative stress. N-acetyl-cysteine, rutin 1 mm and TEMPOL reduced lipoperoxidation in the presence of oxidative stress. However, DHA did not affect lipoperoxidation. At 1 mm, DHA increased DNA damage in the absence of oxidative stress. Dehydroascorbic acid effects could arise from spermatozoa having a low capacity for reducing it to ascorbic acid, and it may be tested in the presence of other antioxidants or reducing power. Future research should focus in testing whether the inhibition of motility observed for NAC, rutin and TEMPOL is reversible. These antioxidants might be useful at lower temperatures (refrigerated storage or cryopreservation) when their protective effects could be advantageous.

Proteomic approach to the study of statin pleiotropy in kidney transplant patients.

BACKGROUND/AIMS: Statins are prescribed in kidney transplant recipients in order to manage dyslipidemia, a common complication in these patients. The efficacy of statins in reducing cholesterol levels has been accompanied by pleiotropic effects. Fifty-four kidney transplant patients were included in the present study, the objective of which was to ascertain the effect of 12 weeks of atorvastatin therapy (10 mg/day) on the patients' lipid profile, renal function, markers of inflammation and plasma peptide profile. METHODS: Biochemical variables were determined with a routine clinical laboratory analyzer, and the proteomic approach was based on magnetic particle-assisted sample processing coupled to mass spectrometry readout. RESULTS: Atorvastatin therapy improved the lipid profile of patients and caused significant changes in their plasma peptide profile; peptides with m/z 1063 and 1898 decreased after treatment and were identified as fragments derived from molecules involved in vascular inflammation, i.e. high-molecular-weight kininogen and complement factor C4, respectively. CONCLUSION: These findings may contribute to the growing body of evidence of the anti-inflammatory actions attributed to statins, by which these drugs could improve these patients' clinical status.

ß-trace protein and cystatin C as predictors of long-term outcomes in patients with acute heart failure.

OBJECTIVES: The purpose of this study was to evaluate the prognostic importance of novel markers of renal dysfunction among patients with acutely destabilized heart failure (ADHF). BACKGROUND: ß-trace protein (BTP) and cystatin C are newer biomarkers for renal dysfunction; the prognostic importance of these tests, particularly BTP, relative to standard measures of renal function remains unclear. METHODS: A total of 220 consecutive hospitalized patients with ADHF were prospectively studied. Blood samples were collected on presentation. In-hospital worsening renal function, as well as mortality and/or heart failure (HF) hospitalization, over a median follow-up period of 500 days was examined as a function of BTP or cystatin C concentrations; results were compared with creatinine, estimated glomerular filtration rate, and blood urea nitrogen. RESULTS: Neither BTP nor cystatin C was associated with worsening renal function during the index hospitalization. A total of 116 patients (53%) either died or were hospitalized for HF during follow-up. Those with adverse outcomes had higher BTP (1.04 mg/l [range 0.80 to 1.49 mg/l] vs. 0.88 mg/l [range 0.68 to 1.17 mg/l], p = 0.003) and cystatin C (1.29 mg/l [range 1.00 to 1.71 mg/l] vs. 1.03 mg/l [range 0.86 to 1.43 mg/l], p = 0.001). After multivariable adjustment, both BTP (hazard ratio: 1.41, 95% confidence interval: 1.06 to 1.88; p = 0.018) and cystatin C (hazard ratio: 1.50, 95% confidence interval: 1.13 to 2.01; p = 0.006) were significant predictors of death/HF hospitalization, whereas serum creatinine, estimated glomerular filtration rate, and blood urea nitrogen were no longer significant. In patients with an estimated glomerular filtration rate >60 ml/min/1.73 m(2), elevated concentrations of BTP and cystatin C were still associated with significantly higher risk of adverse clinical events (p < 0.05). Net reclassification index analysis suggested cystatin C and BTP deliver comparable information regarding prognosis. CONCLUSIONS: Among patients hospitalized with ADHF, BTP and cystatin C predict risk of death and/or HF hospitalization and are superior to standard measures of renal function for this indication.

Effect of nitric oxide on mitochondrial activity of human synovial cells.

BACKGROUND: Nitric oxide (NO) is a messenger implicated in the destruction and inflammation of joint tissues. Cartilage and synovial membrane from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) have high levels of NO. NO is known to modulate various cellular pathways and, thus, inhibit the activity of the mitochondrial respiratory chain (MRC) of chondrocytes and induce the generation of reactive oxygen species (ROS) and cell death in multiple cell types. For these reasons, and because of the importance of the synovial membrane in development of OA pathology, we investigated the effects of NO on survival, mitochondrial function, and activity of fibroblastic human OA synovial cells. METHODS: Human OA synovia were obtained from eight patients undergoing hip joint replacement. Sodium nitroprusside (SNP) was used as a NO donor compound and cell viability was evaluated by MTT assays. Mitochondrial function was evaluated by analyzing the mitochondrial membrane potential (Δψm) with flow cytometry using the fluorofore DePsipher. ATP levels were measured by luminescence assays, and the activities of the respiratory chain complexes (complex I: NADH CoQ1 reductase, complex II: succinate dehydrogenase, complex III: ubiquinol-cytochrome c reductase, complex IV: cytochrome c oxidase) and citrate synthase (CS) were measured by enzymatic assay. Protein expression analyses were performed by western blot. RESULTS: SNP at a concentration of 0.5 mM induced cell death, shown by the MTT method at different time points. The percentages of viable cells at 24, 48 and 72 hours were 86.11 ± 4.9%, 74.31 ± 3.35%, and 43.88 ± 1.43%, respectively, compared to the basal level of 100% (*p < 0.05). SNP at 0.5 mM induced depolarization of the mitochondrial membrane at 12 hours with a decrease in the ratio of polarized cells (basal = 2.48 ± 0.28; SNP 0.5 mM = 1.57 ± 0.11; *p < 0.01). The time course analyses of treatment with SNP at 0.5 mM demonstrated that treatment reliably and significantly reduced intracellular ATP production (68.34 ± 14.3% vs. basal = 100% at 6 hours; *p < 0.05). The analysis of the MRC at 48 hours showed that SNP at 0.5 mM increased the activity of complexes I (basal = 36.47 ± 3.92 mol/min/mg protein, SNP 0.5 mM = 58.08 ± 6.46 mol/min/mg protein; *p < 0.05) and III (basal = 63.87 ± 6.93 mol/min/mg protein, SNP 0.5 mM = 109.15 ± 30.37 mol/min/mg protein; *p < 0.05) but reduced CS activity (basal = 105.06 ± 10.72 mol/min/mg protein, SNP at 0.5 mM = 66.88 ± 6.08 mol/min/mg protein.; *p < 0.05), indicating a decrease in mitochondrial mass. Finally, SNP regulated the expression of proteins related to the cellular cycle; the NO donor decreased bcl-2, mcl-1 and procaspase-3 protein expression. CONCLUSIONS: This study suggests that NO reduces the survival of OA synoviocytes by regulating mitochondrial functionality, as well as the proteins controlling the cell cycle.