Production Of Species Research Articles

Abstract 11457: Overexpression of Thioredoxin-1 Ameliorates the Progression of Diabetic Cardiomyopathy by Preservation of Cardiac Function, Increased Survivability and Decreased Fibrosis in Aged Transgenic Mice

Introduction: Increased production of reactive oxygen species contributes to the etiology of diabetic complications. It acts as an intracellular antioxidant by scavenging reactive oxygen species (ROS) and protect the cells against oxidative stress and cell death. Hypothesis: To determine the long-term effect of streptozotocin (STZ) induced type 1 diabetes on aging Trx-1 transgenic mice related to survivability, heart function, fibrosis, and apoptosis up to 180 days (d) compared to its corresponding wild-type (WT C57Bl/6J). Method: WT and Trx-1 overexpressed mice (Trx-1 Tg/+ ) (8-12 weeks old) were injected with STZ (i.p. 50mg/kg.body weight(bw)-male; 75mg/kg.bw-female for 5 consecutive days) for induction of diabetes (Dia). Fasting blood sugar level (FBS) was measured 30d after the final injection, and FBS >220mg/dL is used as the cut-off for diabetes. Echocardiography, Survival, FBS measurements were performed on 30, 60, 90, 120, 150, and 180d. Immunohistochemistry was performed on heart tissues (30, 90, and 180d). Results: Dia-Trx-1 Tg/+ mice maintained EF at 60d (61.14±2.3%), 120d (56.1±1.94%) and 180d (60.8±4.3%), when compared to 30d (64.4±1.94%) representing preserved cardiac functions. On the other hand, Dia-WT mice show significant progressive depression of EF at 60d (53.8±1.9%), 120d (48.8±2.1%), and 180d (48.9±1.5%), (p<0.05) when compared to 30d (68.4±6.1%) Dia-WT. Moreover, Dia-Trx-1 Tg/+ mice show significantly higher EF than their corresponding Dia-WT counterparts at each time point measured after diabetic induction (n=8-12, p<0.05). Dia-Trx-1 Tg/+ mice (n=71) also exhibited increased survival rate over Dia-WT (n=47) counterparts with a median survival of 150 days compared to 60 days (Log Rank test: p=0.004). Dia-Trx-1 Tg/+ mice also showed a significant reduction in cardiac fibrosis (4.18±1.08% vs. 17.5±5.1%; p=0.03, n=4) and TUNEL positive cells (3.81±1.49 vs. 7.66±1.46; p=0.03, n=4) when compared to their counterpart (Dia-WT) after 180d of diabetes. Conclusion: These results show that Trx-1 Tg/+ mice preserved cardiac functions, increased survival, decreased apoptosis and fibrosis up to 180d under diabetic conditions, and hence suggesting Trx-1 based treatment strategy could prevent heart failure in aging.

Abstract 13283: Mitochondrial DNA Damage and Remodelling of Nucleoid Bodies in Pulmonary Artery Smooth Muscle Cells From Human Patients With Pulmonary Arterial Hypertension

Introduction: Pulmonary arterial hypertension (PAH) is an obstructive pulmonary vasculopathy characterized by pulmonary artery smooth muscle cell (PASMC) hyperproliferation. PAH-PASMC have mitochondrial defects, including increased fragmentation, and increased production of reactive oxygen species (ROS). Mitochondrial-derived ROS can cause DNA damage. Mitochondrial DNA (mtDNA) is surrounded by replication proteins which form nucleoid bodies (NB). Hypothesis: We hypothesize that mtDNA damage is increased and NB structure altered in human PAH PASMC. Methods & Results: We profiled mtDNA and NB composition and integrity in control and PAH PASMC (n = 5/group). mtDNA content was assessed via confocal immunofluorescence of discrete extranuclear double-stranded DNA colocalized with mitochondria. Lower mtDNA content was observed in PAH PASMC vs control (1.8±0.2 vs 3.5±0.2 mtDNAs/μm 3 mitochondria, p<0.0001). Reduced mtDNA in PAH vs control was validated by qPCR (0.69±0.07 vs 1.00±0.07 ng mtDNA/μg cellular protein, n=4-5, p=0.0182). siRNA targeting of Mitofusin2 (Mfn2) in normal PASMC to mimic the mitochondrial fragmentation of PAH, reduced total mtDNA compared to control (1.05±0.06 vs 3.5±0.2 mtDNA/μm 3 mitochondria, n=9, p<0.0001), suggesting increased fission reduces mtDNA expression. Amplification of an 8.8kb mtDNA fragment, a measure of mtDNA damage, revealed increased damage in PAH vs control (14.3±3.9 vs 28.7±2.4 pg mtDNA, n=4, p=0.02). PAH PASMC exhibited higher expression of NB proteins vs control (n=5/measurement): TFAM (+96.7±23.3%, p=0.0032), SSBP (+63.4±14.4%, p=0.0023), and POLG (+59.0±24.0%, p=0.0397). These results were confirmed via immunoblotting for TFAM (+161.9±39.3%, n=4-5, p=0.0045) . Immunofluorescence verified increased TFAM in PAH PASMC vs control. Conclusions: We report a paradoxical upregulation of NB proteins despite reduced mtDNA content and elevated mtDNA damage in PAH. We speculate there is an unsuccessful compensatory attempt to restore mtDNA levels by increasing the expression of mtDNA’s replication apparatus in PAH PASMC.

Abstract 10163: Ca 2+ Within the Mitochondria Can Affect the Occurrence of Triggered Arrhythmias in Rats with Right Ventricular Hypertrophy

Background: Ca 2+ within the sarcoplasmic reticulum affects the propagation of intracellular Ca 2+ (Ca i ) waves, and the velocity of Ca i waves play important roles in the occurrence of triggered arrhythmias. It is still unknown, however, whether Ca 2+ within the mitochondria (Ca m ) affects the velocity of Ca i wave and the occurrence of arrhythmias in right ventricular (RV) hypertrophy. Objective: To know the roles of Ca m in the velocity of Ca i waves and the occurrence of triggered arrhythmias in rats with RV hypertrophy. Methods: Rats were subcutaneously injected with 60 mg/kg monocrotaline (MCT-rats) or solvent (Ctr-rats). Four weeks after the injection, trabeculae were dissected from the RVs. The force was measured using a silicon strain gauge, Ca m using rhod-2, Ca i using microinjected fura-2 and a CCD camera, and production of reactive oxygen species (ROS) from the slope DCF fluorescence during electrical stimulation. Ca 2+ waves and arrhythmias were induced by electrical stimulation (24°C). Results: MCT-rats showed a higher RV pressure, a larger weight ratio of RV free wall to left ventricle (RV/LV), and faster and larger Ca i waves than Ctr-rats (p<0.01). Among MCT-rats, Ru360 (10 μM), a mitochondrial calcium uniporter (MCU) inhibitor, decreased the occurrence of arrhythmias in trabeculae from rats with a higher RV pressure and a larger RV/LV (n=19). Ru360 decreased rhod-2 signal within trabeculae depending on the RV/LV (r=-0.85, p<0.01, n=10) without changes in Ca i transients, suggesting that inhibition of MCU decreases Ca m in MCT-rats with more severe RV hypertrophy. In addition, Ru360 decreased the velocity of Ca i waves depending on systolic RV pressure (r=-0.60, n=6) and the RV/LV (r=-0.64, n= 7), suggesting that inhibition of MCU decreased the velocity of Ca i waves in MCT-rats with higher RV pressure and more severe RV hypertrophy. Ru360 decreased the slope of DCF fluorescence (p<0.01, n=13). Conclusions: In the hearts with severe RV hypertrophy, inhibition of MCU can cause larger reduction of Ca m , decrease ROS production, and decrease the velocity of Ca i waves and arrhythmias. Thus, in the hearts with severe RV hypertrophy, more Ca 2+ may enter the mitochondria through the MCU, and higher Ca m may be involved in the occurrence of arrhythmias.

Abstract 11752: Pitavastatin Treatment Ameliorates HIV-Nef Containing Extracellular Vesicle-Mediated Cardiomyocyte Dysfunction

Rationale: The prevalence of cardiovascular diseases such as heart failure and left ventricular diastolic dysfunction remains high in People Living With HIV (PLWH) even with the advent of modern anti-retroviral therapy (ART). Sustained plasma persistence of the pro-inflammatory HIV protein Nef within extracellular vesicles (EV) of ART-treated patients may contribute to continued disease progression. We hypothesized that HIV-Nef induces cardiomyocyte dysfunction by elevating Reactive Oxygen Species (ROS) production which then increases apoptosis and senescence in cardiomyocytes. Results: Network analysis using Metacore of cytosolic and nuclear proteomics of human cardiomyocytes stimulated with HIV-Nef EV demonstrated changes in metabolic processes, DNA damage, oxidative stress and cytoskeletal remodeling. HIV-Nef EV promotes cardiomyocyte cellular dysfunction as evidenced by increased ROS production, DNA damage, apoptosis and premature senescence in both AC-16 cardiomyocyte cell line (n=5, p<0.001) and iPSC-derived cardiomyocytes (n=4, p<0.01). HIV-Nef EV stimulation induced cardiomyocyte metabolic switch by increasing consumption of anaerobic substrate (Lactic Acid) and decreasing consumption of TCA substrate (Citric Acid). [AE1] Network-science powered pathway analysis of HIV-Nef EV stimulated cardiomyocytes identified a key hub in Rac1-GTPase mediated pathways. Proteomic analysis of HIV-Nef transfected HEK293T cells indicated pitavastatin treatment altered Rac1 trafficking to inhibit interaction with HIV-Nef, suggesting a potential protective mechanism. Subsequently, cardiomyocyte dysfunction was abrogated by clinically relevant doses of pitavastatin (10-100nM ,n=4, p<0.01). Addition of HIV-Nef EV supernatant to AC-16 cardiomyocytes but not to those treated with pitavastatin increased chemotaxis of primary human macrophages (n=4, p<0.05). Conclusion: Pitavastatin treatment may prevent Nef EV-mediated cardiac dysfunction and inflammation in HIV patients. [AE1]I suggest including numbers of technical or biological replicates and p values when possible.

Abstract 9481: Inhibition of Apoptosis and Ferroptosis Signaling Pathways Alleviates Myocardial Ischemia-Reperfusion Injury in Rats Through Modulation of Mitochondrial Function

Introduction: Apoptosis and ferroptosis have been implicated in cardiac ischemia/reperfusion (I/R) injury. Inhibitors of apoptosis (Z-VAD) and ferroptosis (FER-1) have been shown to protect the heart in several cardiovascular disorders. However, the impacts of Z-VAD, FER-1, and combined therapy during cardiac I/R injury have not been elucidated. Hypothesis: Z-VAD and FER-1 provide cardioprotection by reducing myocardial cell injury and improving cardiac function. The combined treatment exerts better benefits than either single regimen. Methods: Male rats (n=25) were equally divided into 5 groups: sham, vehicle, Z-VAD at 3.3 mg/kg, FER-1 at 2 mg/kg, and combined FER-1 and Z-VAD. All treatments were intravenously injected to the rats 15 min before ischemia. Left anterior descending coronary was ligated for 30 min, followed by 120-min reperfusion in all animals except the sham group. Left ventricular ejection fraction (LVEF), infarct size, mitochondrial reactive oxygen species (ROS) production, and protein expressions of apoptosis and ferroptosis were determined. Results: Cardiac I/R injury led to cardiac mitochondrial dysfunction and cell death, contributed to infarct size expansion and LV dysfunction (Fig 1A-D). Z-VAD, FER-1 and combined treatments markedly increased %LVEF, reduced ROS levels and infarct size (the %reduction is 42.9%, 41.4%, and 47.7%, respectively, compared with vehicle). Cleaved caspase 3 (apoptosis) and ACSL4 (ferroptosis) expression levels were also reduced in those rats. The combined treatment did not show a significant advantage, compared with either single regimen (Fig 1A-D). Conclusions: Inhibitors of either apoptosis or ferroptosis similarly reduced cardiac mitochondrial dysfunction, apoptosis and ferroptosis, resulting in decreased infarct size, and finally leading to improved LV function. These findings indicate that ferroptosis could be a novel therapeutic target for cardioprotection in cardiac I/R injury.

Abstract 13389: Critical Role of Electron Leak From Mitochondrial Electron Transport Chain Complex I in Oxygen Sensing in the Rabbit Ductus Arteriosus

Introduction: The ductus arteriosus (DA) allows placentally oxygenated blood to bypass the non-ventilated fetal lungs and enter the systemic circulation. The fetus’ systemic pO 2 is ~40 mmHg, maintaining the DA in a state of hypoxic vasodilation. As pO 2 rises, with the first breath, the DA constricts and achieves functional closure within minutes. We have identified a mitochondrial O 2 -sensor controlling functional closure within DA smooth muscle cells (DASMC). Increased pO 2 leads to increased reactive oxygen species (ROS) production by complexes I and III of the electron transport chain (ETC). These ROS are diffusible signaling molecules and inhibit redox sensitive potassium channels causing DASMC depolarization and vasoconstriction. We propose this reflects electron leak from ETC complexes. Using the term rabbit model, we investigated the differential function of ETC I vs III complex-specific electron leak suppressors (S1QEL vs S3QEL) on vascular tone in DA rings. Methods: DA’s from term rabbits (n=11 rabbits, n=29 individual DA rings) were mounted on a force transducer at ~800 mg in Krebs at a PO 2 of ~40mmHg. DA’s were subject to varying pO 2 levels (from 35-155 mmHg) and at peak O 2 constriction were treated with ETC leak suppressors, ETC complex inhibitors or vehicle controls (DMSO). Results: Exposure to normoxia caused an average 1100±62 mg increase in DA tension. S1QEL (50 μM) caused an average 20.9±6.2% reduction in DA tension (n=8) whereas S3QEL (100 μM) only reduced DA tension by 4.4±2.2% (n=4). Rotenone (complex I inhibitor) and antimycin A (complex III inhibitor) also relaxed the DA while vehicle had minimal effect (-2.6±1.5%). Conclusions: Electron leak from complex 1, but not complex 3, is implicated in DA O 2 -sensing.

Simulation of Wood Combustion in PATO Using a Detailed Pyrolysis Model Coupled to fireFoam

The numerical simulation of fire propagation requires capturing the coupling between wood pyrolysis, which leads to the production of various gaseous species, and the combustion of these species in the flame, which produces the energy that sustains the pyrolysis process. Experimental and numerical works of the fire community are targeted towards improving the description of the pyrolysis process to better predict the rate of production and the chemical nature of the pyrolysis gases. We know that wood pyrolysis leads to the production of a large variety of chemical species: water, methane, propane, carbon monoxide and dioxide, phenol, cresol, hydrogen, etc. With the idea of being able to capitalize on such developments to study more accurately the physics of fire propagation, we have developed a numerical framework that couples a detailed three-dimensional pyrolysis model and fireFoam. In this article, we illustrate the capability of the simulation tool by treating the combustion of a wood log. Wood is considered to be composed of three phases (cellulose, hemicellulose and lignin), each undergoing parallel degradation processes leading to the production of methane and hydrogen. We chose to simplify the gas mixture for this first proof of concept of the coupling of a multi-species pyrolysis process and a flame. In the flame, we consider two separate finite-rate combustion reactions for methane and hydrogen. The flame evolves during the simulation according to the concentration of the two gaseous species produced from the material. It appears that introducing different pyrolysis species impacts the temperature and behavior of the flame.

Photoactivated cell-killing amino-based flavylium compounds

Photodynamic therapy (PDT) is a well-established therapeutic for the treatment of different diseases. The growing interest of this technique required the development of new photosensitizers with better photo-features. This work reports the study of the potential of five nature-inspired amino-based flavylium compounds with different structural features as photosensitizers towards topical PDT. In terms of dark cytotoxicity the five pigments were tested towards confluent skin cells in both fibroblasts and keratinocytes. In the range of concentrations tested (6.3–100 μM), keratinocytes were more prone to growth inhibition and the IC50 values for 5OH4′NMe2, 7NEt2st4′NMe2 and 7NEt24′NH2 were determined to be 47.3 ± 0.3 μM; 91.0 ± 0.8 μM and 29.8 ± 0.8 μM, respectively. 7NEt24′NMe2, 7NEt2st4′NMe2 and 7NEt24′NH2 showed significant fluorescence quantum yields (from 3.40 to 20.20%) and production of singlet oxygen (1O2). These latter chromophores presented IC50 values of growth inhibition of keratinocytes between 0.9 and 1.5 µM, after 10 min of photoactivation with white light. This cellular damage in keratinocyte cells upon white light activation was accompanied with the production of reactive oxygen species (ROS). It was also found that the compounds can induce damage by either type I (ROS production) or type II (singlet oxygen) PDT mechanism, although a higher cell survival was observed in the presence of 1O2 quenchers. Overall, a structure–activity relationship could be established, ranking the most important functional groups for the photoactivation efficiency as follows: C7-diethylamino > C4′-dimethylamino > C2-styryl.

Silencing of vitellogenin gene contributes to the promise of controlling red palm weevil, Rhynchophorus ferrugineus (Olivier)

Red palm weevil [Rhynchophorus ferrugineus (Olivier)], is native to South Asia and expanding its distribution range globally. Recent invasions of red palm weevil around the world, including Saudi Arabia, has become a global constraint for the production of palm species. Although, several control measures have been tested, none of them seemed successful against this invasive species. Therefore, we focused on silencing the reproduction control gene vitellogenin (Vg) based on RNA interference (RNAi) strategy for its possible application in the management of R. ferrugineus. The Vg is a major yolk protein precursor critical for oogenesis. To do this, fat body transcriptome of R. ferrugineus female adults was sequenced, which provided partial Vg gene transcript (FPKM 5731.60). A complete RfVg gene transcript of 5504 bp encoding 1787 amino acids was then sequenced using RCAE-PCR strategy and characterized. Phylogenetic analysis suggested that RfVg has closer ancestry to the coleopteran insects. The RfVg-based RNAi significantly suppressed the expressions of Vg gene. The 15, 20 and 25 days post-injection periods suppressed Vg expressions by 95, 96.6 and 99%, respectively. The suppressed Vg expressions resulted in the dramatic failure of Vg protein expression, which caused atrophied ovaries or no oogenesis and ultimately eggs were not hatched. These results suggest that knockdown of Vg gene involved in R. ferrugineus reproduction is a promising target for RNAi-based management of R. ferrugineus.

Characterization of Biogas-Syngas Turbulent MILD ‎Combustion in the Jet in Hot Co-Flow Burner

Moderate or Intense Low–oxygen Diluted (MILD) combustion is a promising technology with interesting ‎properties such as high efficiency and zero-emission. The biogas-syngas mixture is also considered a ‎promising new renewable biofuel with low emissions. This work aims to examine the effects of several ‎parameters on the biogas-syngas flame structure and emissions under MILD conditions in the Jet in Hot ‎Co flow (JHC) burner. The turbulence is modeled by the modified standard k-ε model; whereas ‎combustion-turbulence interaction is handled by the Eddy Dissipation Concept (EDC) in conjunction with ‎three detailed reaction mechanisms, namely: GRI-Mech 3.0, GRI-Mech 2.11, and DRM 2.11. Effects of ‎biogas-syngas composition, temperature, and oxygen concentration in the hot co-flow and Reynolds ‎number of the fuel jet have been elucidated. Results show that flame structure is more sensitive to the ‎increase of hydrogen in syngas than that of methane in biogas. An increase of oxygen concentration or ‎temperature in the co-flow stream leads to more NO formation whereas Reynolds number augmentation ‎reduced them. Furthermore, NO species production is globally governed by the NNH route‎‎.

Increased Post-Hypoxic Oxidative Stress and Activation of the PERK Branch of the UPR in Trap1-Deficient Drosophila melanogaster Is Abrogated by Metformin

Hypoxia is known to impair mitochondrial and endoplasmic reticulum (ER) homeostasis. Post-hypoxic perturbations of the ER proteostasis result in the accumulation of misfolded/unfolded proteins leading to the activation of the Unfolded Protein Response (UPR). Mitochondrial chaperone TNF receptor-associated protein 1 (TRAP1) is reported to preserve mitochondrial membrane potential and to impede reactive oxygen species (ROS) production thereby protecting cells from ER stress as well as oxidative stress. The first-line antidiabetic drug Metformin has been attributed a neuroprotective role after hypoxia. Interestingly, Metformin has been reported to rescue mitochondrial deficits in fibroblasts derived from a patient carrying a homozygous TRAP1 loss-of-function mutation. We sought to investigate a putative link between Metformin, TRAP1, and the UPR after hypoxia. We assessed post-hypoxic/reperfusion longevity, mortality, negative geotaxis, ROS production, metabolic activity, gene expression of antioxidant proteins, and activation of the UPR in Trap1-deficient flies. Following hypoxia, Trap1 deficiency caused higher mortality and greater impairments in negative geotaxis compared to controls. Similarly, post-hypoxic production of ROS and UPR activation was significantly higher in Trap1-deficient compared to control flies. Metformin counteracted the deleterious effects of hypoxia in Trap1-deficient flies but had no protective effect in wild-type flies. We provide evidence that TRAP1 is crucially involved in the post-hypoxic regulation of mitochondrial/ER stress and the activation of the UPR. Metformin appears to rescue Trap1-deficiency after hypoxia mitigating ROS production and downregulating the pro-apoptotic PERK (protein kinase R-like ER kinase) arm of the UPR.

Supplementation of Beltsville extender with quercetin improves the quality of frozen-thawed rooster semen

ABSTRACT 1. Excessive reactive oxygen species (ROS) production during the sperm freeze-thawing process leads to membrane lipid peroxidation, DNA damage, and motility loss. 2. This study examined the effect of supplementation of Beltsville poultry semen extender with different concentrations of quercetin (and antioxidants) on the cryopreservation of rooster sperm. 3. Semen samples were collected from six Ross broiler breeders via abdominal massage twice a week for 4 weeks (eight replicates), and were divided into five equal aliquots to be diluted in Beltsville extenders that contained different concentrations of quercetin: 0, 5, 10, 15, and 20 mM. Motility, membrane functionality, abnormal morphology, lipid peroxidation, mitochondrial activity, viability, apoptosis status, and fertility potential were assessed post thaw. 4. The addition of 10 and 15 mM quercetin to the semen extender significantly increased the total motility, straight-line velocity (VSL), and sperm membrane functionality compared with the other groups (P ≤ 0.05). Moreover, 10 mM quercetin caused higher progressive motility (34.86 ± 3.80%), curvilinear velocity (VCL; 175.11 ± 3.20 µm/s), average path velocity (VAP; 44.35 ± 11.06 µm/s), viability (59.14 ± 1.36%), mitochondrial activity (80.14 ± 2.07%), lower abnormal morphology (19.21 ± 0.45%), and lower lipid peroxidation (2.7 ± 0.13 nmol/ml) compared with the other groups (P < 0.05). The rate of fertility and hatchability after artificial insemination was not affected by experimental groups. 5. In conclusion, supplementation of Beltsville extender with 10 mM quercetin could be a suitable method to improve post-thawed rooster sperm quality resulting in better freeze/thaw characteristics.

Targeting the complex I and III of mitochondrial electron transport chain as a potentially viable option in liver cancer management

Liver cancer is one of the most common and lethal types of oncological disease in the world, with limited treatment options. New treatment modalities are desperately needed, but their development is hampered by a lack of insight into the underlying molecular mechanisms of disease. It is clear that metabolic reprogramming in mitochondrial function is intimately linked to the liver cancer process, prompting the possibility to explore mitochondrial biochemistry as a potential therapeutic target. Here we report that depletion of mitochondrial DNA, pharmacologic inhibition of mitochondrial electron transport chain (mETC) complex I/complex III, or genetic of mETC complex I restricts cancer cell growth and clonogenicity in various preclinical models of liver cancer, including cell lines, mouse liver organoids, and murine xenografts. The restriction is linked to the production of reactive oxygen species, apoptosis induction and reduced ATP generation. As a result, our findings suggest that the mETC compartment of mitochondria could be a potential therapeutic target in liver cancer.

Overview of Multiple Applications of Basil Species and Cultivars and the Effects of Production Environmental Parameters on Yields and Secondary Metabolites in Hydroponic Systems

Basil (Ocimum basilicum L.), including other species and cultivars, is an excellent source of nutritional compounds, the accumulation of which can be stimulated by exogenous factors (environmental and nutritional conditions). Although best practices are relatively established for mature basil plants, microgreens production requires further research to optimize quality and quantity. The study objectives are (i) to provide an overview of the many uses of basil, (ii) collate and present common hydroponic systems available in the market, (iii) review effects of key production environment parameters on basil yields in hydroponic systems, and (iv) summarize the effects of the growth environments on yield quantity and quality of basil microgreens. The paper analyzes in detail key production parameters of basil microgreens in hydroponic systems, such as temperature, humidity, pH, electrical conductivity, dissolved oxygen, carbon dioxide, nutrient solutions, and the influence of light (quantity, quality, and photoperiods). The collated literature review has shown that basil, grown hydroponically, can tolerate high variations of environmental parameters: pH 5.1–8.5, temperature 15–24 °C, relative humidity 60–70%, electrical conductivity up to 1.2 mS cm−1, depending on the developmental stage, dissolved oxygen at 4 mg L−1 (optimally 6.5 mg L−1), and light intensity between 200 and 400 μmol m−2 s−1. The study has synthesized an overview of different production parameters to provide guidance on the optimization of environmental conditions to ensure the quantity and quality production of basil microgreens. Improving the quality of basil microgreens can ideally spur continued gastronomic interest in microgreens in general, which will encourage more entrepreneurs to grow basil and other microgreens. Hence, the study findings are a great resource to learn about the effects of different environments on basil microgreen production. This information can inform research for successful production of different species and cultivars of basil microgreens, and establishing testing protocols to improve the quantity and quality of the harvest.

Activation of adenosine receptors improved cardiac dysfunction induced by myocardial infarction

Abstract Introduction Acute myocardial infarction (MI) is characterized by an intense inflammatory response leading to cardiac remodeling and subsequent cardiac dysfunction. Purpose Activation of adenosine A2A and A3 receptors induced by LASSBio-1027 could improve cardiac function, through the reduction of the inflammatory response and the activated cardiac fibroblasts (CF). Methods The Animal Care and Use Committee approved the experimental protocols. MI was induced in male Wistar rats by ligation of the anterior descending coronary artery and confirmed by transthoracic echocardiography. The experimental groups were Sham and MI treated orally with either vehicle or LASSBio-1027 (30 or 70 μmol/kg) during 7 days. At the endpoint, hemodynamic parameters were evaluated and hearts were prepared for histological and immunohistochemical analysis for fibrosis and cellular infiltrate detection and for α-SMA, TNF-α, iNOs, c-fos, p38MAPK expression, respectively. Production of reactive oxygen species and collagen in CF with H2O2–induced oxidative stress was observed in the presence or absence of LASSBio-1027 (10 μM). Results Diastolic dysfunction was detected in MI group because the filling pressure (E/e') and the left ventricular end diastolic pressure (LVEDP) were increased from 22.9±1.6 to 37.0±3.7 and from 3.2±0.9 to 18.2±2.2 mmHg (p&amp;lt;0.01), respectively. LASSBio-1027 (70 μmol/kg) prevented diastolic dysfunction since E/e' and LVEDP reduced to 23.8±5.3 and 6.4±1.6 mmHg (p&amp;lt;0.01). Adenosine receptor agonist improved systolic dysfunction in MI group with increase of ejection fraction from of 30.1±3.1 to 48.4±4.8% (p&amp;lt;0.05). As shown in figure 1, MI promoted an increase in collagen deposition, cellular infiltrate, expression of SMA-α, TNF-α, iNOs, c-fos and p38MAPK which confirmed the intense fibrosis, cell proliferation and inflammation. Oral administration of LASSBio-1027 reduced those parameters in a dose dependent manner. LASSBio-1027 reduced H2O2-induced fibroblast-myofibroblast transformation with the decrease of stain density of α-SMA from 1794.4 to 574.4 staining area/cell and collagen production from 33.7±9.6 to 22.1±18.9% (p&amp;lt;0.05). Effects of LASSBio-1027 were inhibited by the antagonists of A2A (ZM-241385) and A3 (MRE3008F20) receptors. Conclusions Activation of adenosine A2A and A3 receptors by LASSBio-1027 improved cardiac function and remodeling induced by MI. Funding Acknowledgement Type of funding sources: Foundation. Main funding source(s): CNPq. CAPES, FAPERJ Histological AnalysisImmunohistochemistry analysis

Prevention of contrast-induced nephropathy with urine alkalinization: the TEATE study

Abstract Background Contrast-induced acute kidney injury (CI-AKI) after coronary angiography and percutaneous interventions (PCI) impacts on hospitalization duration and mortality. Pre-procedural hydration is the sole strategy currently recommended for preventing CI-AKI. The role of sodium bicarbonate (SB) although attractive, since urine alkalinization suppresses the production of reactive oxygen species, is still controversial, and the optimal dosing to attain adequate urine alkalinization is still undefined. Purpose The PrevenTion of contrast-inducEd nephropathy with urine alkalinization (TEATE) study was a prospective 3-center 3-arm single-blind randomized controlled trial testing the hypothesis that adequate urine alkalinization is associated with CI-AKI prevention. Secondary endpoints were the efficacy of SB vs saline in achieving adequate urine alkalinization and reducing the incidence of CI-AKI compared with saline. Methods Patients candidate to coronary angiography and/or PCI with moderate-to-severe chronic kidney disease [eGFR of 15 to 60 mL/min/1.73 m2, by the Modification of Diet in Renal Disease Study equation (MDRD)] were randomly assigned to saline hydration (control), oral SB or i.v. SB. The study protocol was registered. We evaluated urinary pH at the time of hospitalization, immediately before coronary angiography and 24–48 hours after angiography. According to urine pH immediately before the procedure, patients were divided in two groups above or below a pH cut-off of 6. Results We enrolled a total of 241 patients: 81 were randomly assigned to the control group, 82 to i.v. SB and 78 to oral SB. Patients achieving a urinary pH &amp;gt;6 before angiography had a lower incidence of CI-AKI (46%) than patients with urinary pH ≤6 (54%) [OR=0.48 (95% CI 0.25–0.9) p=0.023]. The number of patients with urine pH &amp;gt;6 was higher in both the i.v. (71%) and the oral SB (65%) groups compared to the hydration-only group (44%, p=0.004). We found however no difference in the incidence of CI-AKI in the 3 treatment arms (20% in hydration alone, 21% in oral SB group and 22% in iv SB group) (p=0.94). Subgroup analyses according to basal urine pH and eGFR ranges failed to identify statistically significant differences in the development of CI-AKI according to treatment allocation. Conclusions Urinary pH before the administration of contrast medium is an inverse correlate of CI-AKI incidence, and SB is superior to hydration alone in achieving urinary alkalinization. Since, however, SB did not reduce the incidence of CI-AKI, we conclude that urinary pH is a marker and not a mediator of CI-AKI. Funding Acknowledgement Type of funding sources: None.

Sex-related differential susceptibility to ponatinib-induced cardiotoxicity and its relationship to modulation of Notch signaling in a murine model

Abstract Background Ponatinib (PON), a tyrosine kinase inhibitor approved in chronic myeloid leukemia, has proven cardiovascular toxicity. Although sex is a risk factor for PON-induced cardiotoxicity in humans, little is known about its mechanisms in general, and sex-related mechanisms in particular. Objectives To determine the mechanisms of sex-related PON-induced cardiotoxicity and identify potential rescue strategies in a murine model. Methods 24-months-old male and female C57B5 mice were treated with 3 mg/kg/day of PON or vehicle via oral gavage for 28 days, with/without siRNA-Notch1 or siRNA-scrambled via tail vein every 3 days. Results PON + scrambled siRNA-treated male mice had a higher number of TUNEL-positive cells, a higher percentage of senescence-associated β-galactosidase positive senescent cardiac areas, as well as a lower reactivity degree for the survival marker Bmi1 than female counterparts. Proteomics analysis of cardiac tissue showed upstream activation of nitric oxide synthase (NOS) type 2, downstream activation of cell death and production of reactive oxygen species in PON + scrambled siRNA- compared to vehicle or PON + Notch1 siRNA-treated male mice. Upstream analysis showed beta-estradiol activation, while downstream analysis showed activation of cell survival and inhibition of cell death in PON + scrambled siRNA compared to vehicle-treated female mice. PON + scrambled siRNA-treated mice also showed a downregulation of cardiac actin, which was more marked in male; as well as vessel density, which was more marked in female mice. Female hearts showed a greater extent of cardiac fibrosis than male counterparts at baseline, with no significant changes after PON treatment. In contrast, PON + scrambled siRNA-treated mice had less fibrosis than vehicle or PON + Notch1-siRNA-treated mice. Left ventricular systolic dysfunction shown in PON + scrambled siRNA-treated male mice and - to a lesser extent - by female mice was similarly reversed in both PON + Notch1-siRNA-treated male and female mice (Table 1). Conclusions We found a sex-related differential susceptibility and Notch1 modulation in PON-induced cardiotoxicity. This can improve our understanding of sex-related differences and help identify biomarkers in PON cardiotoxicity. Funding Acknowledgement Type of funding sources: None.

Stamp2 Protects From Maladaptive Structural Remodeling and Systolic Dysfunction in Post-Ischemic Hearts by Attenuating Neutrophil Activation.

The six-transmembrane protein of prostate 2 (Stamp2) acts as an anti-inflammatory protein in macrophages by protecting from overt inflammatory signaling and Stamp2 deficiency accelerates atherosclerosis in mice. Herein, we describe an unexpected role of Stamp2 in polymorphonuclear neutrophils (PMN) and characterize Stamp2’s protective effects in myocardial ischemic injury. In a murine model of ischemia and reperfusion (I/R), echocardiography and histological analyses revealed a pronounced impairment of cardiac function in hearts of Stamp2-deficient- (Stamp2-/- ) mice as compared to wild-type (WT) animals. This difference was driven by aggravated cardiac fibrosis, as augmented fibroblast-to-myofibroblast transdifferentiation was observed which was mediated by activation of the redox-sensitive p38 mitogen-activated protein kinase (p38 MAPK). Furthermore, we observed increased production of reactive oxygen species (ROS) in Stamp2-/- hearts after I/R, which is the likely cause for p38 MAPK activation. Although myocardial macrophage numbers were not affected by Stamp2 deficiency after I/R, augmented myocardial infiltration by polymorphonuclear neutrophils (PMN) was observed, which coincided with enhanced myeloperoxidase (MPO) plasma levels. Primary PMN isolated from Stamp2-/- animals exhibited a proinflammatory phenotype characterized by enhanced nuclear factor (NF)-κB activity and MPO secretion. To prove the critical role of PMN for the observed phenotype after I/R, antibody-mediated PMN depletion was performed in Stamp2 -/- mice which reduced deterioration of LV function and adverse structural remodeling to WT levels. These data indicate a novel role of Stamp2 as an anti-inflammatory regulator of PMN and fibroblast-to-myofibroblast transdifferentiation in myocardial I/R injury.

Effect of Silver Nanoparticles on Blue Light Phototoxicity against Fusobacterium nucleatum

A previous study showed that sub-lethal exposure of blue light caused cell membrane damage in Fusobacterium nucleatum (Fn). The aim of the present study was to test the combined effect of blue light and silver nanoparticles against Fn. Bacterial suspensions were exposed to blue light (400–500 nm) with or without silver nanoparticles (10 nm). Exposed and non-exposed samples were studied for malodor production (Odor judge scores), VSC levels (Halimeter), reactive oxygen species (ROS) production (fluorimeter), and bacterial cell membrane damage (fluorescence microscopy). The results showed that combining blue light exposure and silver nanoparticles significantly reduced malodor and VSC production by Fn concomitant with increased ROS levels and bacterial cell membrane damage. These results suggest that silver nanoparticles may increase blue light phototoxicity against Fn.

Biosafety assessment of Acinetobacter strains isolated from the Three Gorges Reservoir region in nematode Caenorhabditis elegans

Acinetobacter has been frequently detected in backwater areas of the Three Gorges Reservoir (TGR) region. We here employed Caenorhabditis elegans to perform biosafety assessment of Acinetobacter strains isolated from backwater area in the TGR region. Among 21 isolates and 5 reference strains of Acinetobacter, exposure to Acinetobacter strains of AC1, AC15, AC18, AC21, A. baumannii ATCC 19606T, A. junii NH88-14, and A. lwoffii DSM 2403T resulted in significant decrease in locomotion behavior and reduction in lifespan of Caenorhabditis elegans. In nematodes, exposure to Acinetobacter strains of AC1, AC15, AC18, AC21, A. baumannii, A. junii and A. lwoffii also resulted in significant reactive oxygen species (ROS) production. Moreover, exposure to Acinetobacter isolates of AC1, AC15, AC18, and AC21 led to significant increase in expressions of both SOD-3::GFP and some antimicrobial genes (lys-1, spp-12, lys-7, dod-6, spp-1, dod-22, lys-8, and/or F55G11.4) in nematodes. The Acinetobacter isolates of AC1, AC15, AC18, and AC21 had different morphological, biochemical, phylogenetical, and virulence gene properties. Our results suggested that exposure risk of some Acinetobacter strains isolated from the TGR region exists for environmental organisms and human health. In addition, C. elegans is useful to assess biosafety of Acinetobacter isolates from the environment.