μM Concentration Research Articles

Allelopathic studies with furanocoumarins isolated from Ducrosia anethifolia. In vitro and in silico investigations to protect legumes, rice and grain crops

Six different furanocoumarins were isolated from the aerial parts of Ducrosia anethifolia and tested in vitro for plant cell elongation in etiolated wheat coleoptile. They were also tested for their ability to control three different weeds: ribwort plantain, annual ryegrass, and common purslane. These compounds exhibited strong inhibition of plant cell elongation. In the case of (+)-heraclenin, the IC50 was lower than 20 μM, indicating a better inhibition than the positive control Logran®. Computational experiments for docking and molecular dynamics revealed for the investigated furanocoumarins bearing an epoxide moiety an improved fitting and stronger interaction with the auxin-like TIR1 ubiquitin ligase. Furthermore, the formed inhibition complex remained also stable during dynamic evaluation. Bidental interaction at the active site, along with an extended hydrogen-bond lifetime, explained the enhanced activity of the epoxides. The in vitro weed bioassay results showed that Plantago lanceolata was the most affected weed for germination, root, and shoot development. In addition, (+)-heraclenin displayed better inhibition values than positive control even at 300 μM concentration.

Wnt signaling pathway inhibitor promotes mesenchymal stem cells differentiation into cardiac progenitor cells in vitro and improves cardiomyopathy in vivo.

Cardiovascular diseases particularly myocardial infarction (MI) are the leading cause of mortality and morbidity around the globe. As cardiac tissue possesses very limited regeneration potential, therefore use of a potent small molecule, inhibitor Wnt production-4 (IWP-4) for stem cell differentiation into cardiomyocytes could be a promising approach for cardiac regeneration. Wnt pathway inhibitors may help stem cells in their fate determination towards cardiomyogenic lineage and provide better homing and survival of cells in vivo. Mesenchymal stem cells (MSCs) derived from the human umbilical cord have the potential to regenerate cardiac tissue, as they are easy to isolate and possess multilineage differentiation capability. IWP-4 may promote the differentiation of MSCs into the cardiac lineage. To evaluate the cardiac differentiation ability of IWP-4 and its subsequent in vivo effects. Umbilical cord tissue of human origin was utilized to isolate the MSCs which were characterized by their morphology, immunophenotyping of surface markers specific to MSCs, as well as by tri-lineage differentiation capability. Cytotoxicity analysis was performed to identify the optimal concentration of IWP-4. MSCs were treated with 5 μM IWP-4 at two different time intervals. Differentiation of MSCs into cardiomyocytes was evaluated at DNA and protein levels. The MI rat model was developed. IWP-4 treated as well as untreated MSCs were implanted in the MI model, then the cardiac function was analyzed via echocardiography. MSCs were labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) dye for tracking, while the regeneration of infarcted myocardium was examined by histology and immunohistochemistry. MSCs were isolated and characterized. Cytotoxicity analysis showed that IWP-4 was non-cytotoxic at 5 μM concentration. Cardiac specific gene and protein expression analyses exhibited more remarkable results in fourteen days treated group that was eventually selected for in vivo transplantation. Cardiac function was restored in the IWP-4 treated group in comparison to the MI group. Immunohistochemical analysis confirmed the homing of pre-differentiated MSCs that were labeled with DiI cell labeling dye. Histological analysis confirmed the significant reduction in fibrotic area, and improved left ventricular wall thickness in IWP-4 treated MSC group. Treatment of MSCs with IWP-4 inhibits Wnt pathway and promotes cardiac differentiation. These pre-conditioned MSCs transplanted in vivo improved cardiac function by cell homing, survival, and differentiation at the infarcted region, increased left ventricular wall thickness, and reduced infarct size.

Acetylsalicylic acid-sulfa drugs conjugates as potential urease inhibitors and anti-inflammatory agents: bio-oriented drug synthesis, molecular docking, and dynamics simulation studies

To explore the new mode of action and reduce side effects, making conjugates of existing drugs is becoming an attractive tool in the realm of medicinal chemistry. In this work, we exploited this approach and synthesized new conjugates to assess their activities against the enzymes involved in different pathological conditions. Specifically, we design and synthesized conjugates involving acetylsalicylic acid and sulfa drugs, validating the newly crafted conjugates using techniques like IR, 1HNMR, 13CNMR, and elemental analysis. These conjugates underwent assessment for their ability to inhibit cyclooxygenase-2 (COX-2), urease enzymes, and their anti-inflammatory potential. A competitive mode of urease inhibition was observed for acetylsalicylic acid conjugated with sulfanilamide, sulfacetamide, and sulfadiazine with IC50 of 2.49 ± 0.35 µM, 6.21 ± 0.28 µM, and 6.57 ± 0.44 µM, respectively. Remarkably, the acetylsalicylic acid-sulfamethoxazole conjugate exhibited exceptional anti-inflammatory activity, effectively curtailing induced edema by 83.7%, a result akin to the reference anti-inflammatory drug indomethacin’s performance (86.8%). Additionally, it demonstrated comparable COX-2 inhibition (75.8%) to the reference selective COX-2 inhibitor celecoxib that exhibited 77.1% inhibition at 10 µM concentration. To deepen our understanding, we employed molecular docking techniques to predict the binding interactions of competitive inhibitors with COX-2 and urease receptors. Additionally, MD simulations were carried out, confirming the stability of inhibitor-target complexes throughout the simulation period, devoid of significant conformational changes. Collectively, our research underscores the potential of coupling approved medicinal compounds to usher in novel categories of pharmacological agents, holding promise for addressing a wide spectrum of pathological disorders involving COX-2 and urease enzymes. Communicated by Ramaswamy H. Sarma

Flavonoids as potential reactivators of structural mutation p53Y220C by computational and cell-based studies

The p53 Y220C is one of the most frequently observed structural mutants in various human cancers. The substitution of residue Tyr to Cys makes the p53 DNA binding domain susceptible to solvent entry into the hydrophobic core of the domain thereby destabilizing p53, which results in loss of its tumor suppressor activity. The mutation creates a structural crevice at the region between S3/S4 and S7/S8 loops in the DNA binding domain which can be targeted by small molecules. Studies have shown that the synthetic and natural compounds could bind to this crevice and restore the structure and function of the mutant p53Y220C to the wild type. In our previous study, we have shown Curcumin could rescue the function of mutant p53Y220C in pancreatic cancer cell line BxPC-3 harboring genomic mutation. In this study, we explored six flavonoids structurally similar to Curcumin such as Apigenin, Isoliquiritigenin, Liquiritigenin, Luteolin, Methylophiopogonanone A (MPA), and Methylophiopogonanone B (MPB) to test their potency to restore p53Y220C by molecular docking, molecular dynamics simulations and cytotoxicity assay. The secondary structure analysis after the MD simulations suggested that these compounds could stabilize the mutant p53 DNA binding domain to the wild type. In the cell-based cytotoxicity studies using p53Y220C harbouring BxPC-3 cell lines, the compounds MPA and MPB showed 75% cell death at 100 µM concentration. We proposed that the flavonoids MPA and MPB have the therapeutic potential to restore p53Y220C and could be used as a combinatorial therapy to reduce the dosage burden. Communicated by Ramaswamy H. Sarma

Short-term exposure to reduced specific-size ambient particulate matter increase the risk of cause-specific cardiovascular disease: A national-wide evidence from hospital admissions

Evidence for the health effects of ambient PM1 (particulate matter with an aerodynamic diameter ≤ 1 µm) pollution is limited, and it remains unclear whether a smaller particulate matter has a greater impact on human health. We conducted a time-series study in 184 major cities by extracting daily hospital data on admissions for ischemic heart disease, heart failure, heart rhythm disturbances, and stroke between 2014 and 2017 from a medical insurance claims database of 0.28 billion beneficiaries. City-specific associations were estimated with over-dispersed generalized additive models. A random-effects meta-analysis was used to estimate regional and national average associations. We conducted stratified and meta-regression analyses to explore potential effect modifiers of the association. We recorded 8.83 million cardiovascular admissions during the study period. At the national-average level, a 10-μg/m3 increase in same-day PM1, PM2.5(particulate matter with an aerodynamic diameter ≤ 2.5 µm) and PM10(particulate matter with an aerodynamic diameter ≤ 10 µm) concentrations corresponded to a 1.14% (95% confidence interval 0.88–1.41%), 0.55% (0.40–0.70%), and 0.45% (0.36–0.55%) increase in cardiovascular admissions, respectively. PM1 exposure was also positively associated with all cardiovascular disease subtypes, including ischemic heart disease (1.28% change; 0.99–1.56%), heart failure (1.30% change; 0.70–1.91%), heart rhythm disturbances (1.11% change; 0.65–1.58%), and ischemic stroke (1.29% change; 0.88–1.71%). The associations between PM1 and cardiovascular admissions were stronger in cities with lower PM1 levels, higher air temperatures and relative humidity, as well as in subgroups with elder age (all P < 0.05). This study provides robust evidence of short-term associations between PM1 concentrations and increased hospital admissions for all major cardiovascular diseases in China. Our findings suggest a greater short-term impact on cardiovascular risk from PM1 in comparison to PM2.5 and PM10.

Novel azole-urea hybrids as VEGFR-2 inhibitors: Synthesis, in vitro antiproliferative evaluation and in silico studies

The vascular endothelial growth factor receptor-2 (VEGFR-2) is a receptor tyrosine kinase known to be abnormally expressed in various malignant tumors, including breast cancer, and is considered one of the most important contributors to tumor angiogenesis. Sorafenib is one of many VEGFR-2 inhibitors that have received approval for clinical use from the US FDA in recent years. Accordingly, in this study, the synthesis of two new pyrazoles, six 1,3,4-oxadiazoles, four 1,3,4-thiadiazoles, and ten 1,2,4-triazole-3-thione derivatives having structural characteristics similar to sorafenib was carried out. A preliminary screening of synthesized compounds and known inhibitors sorafenib and staurosporine at 10 µM concentration on in vitro activity of VEGFR-2 was performed, and compounds 10c, 8a, and 11 g were identified as the most potent derivatives with% VEGFR-2 residual activities lower than 30%, and dose-dependent inhibition studies was carried out to determine the IC50 values of these inhibitors. Compound 10c was found to be the most potent inhibitor of VEGFR-2 activity with an IC50 value of 0.664 µM. The anti-proliferative activity of synthesized derivatives was assessed against a breast carcinoma (MCF-7) cell line, a triple negative human breast adenocarcinoma (MDA-MB-231) cell line, and noncancerous fibroblast cells (L929). Compound 8a displayed superior activity when compared to sorafenib against MCF-7 (7.69 fold) and MDA-MB-231 (1.52 fold) cell lines while displaying 3.75-fold less toxicity against the normal L929 cell line. Annexin V binding assay revealed that compound 8a significantly increased early and late apoptosis in MCF-7 cells and late apoptosis and necrosis in MDA-MB-231 cells. Computational studies such as molecular docking and ADMET evaluation were performed to elucidate the binding interactions and drug-likeness of the synthesized compounds. The results indicate that compound 8a could be a promising candidate for the development of a novel anti-angiogenic and anti-proliferative agent.

Toxic effects of benfluralin on zebrafish embryogenesis via the accumulation of reactive oxygen species and apoptosis

The dinitroaniline herbicide benfluralin is used weed control in conventional systems and poses a high risk of accumulation in aquatic systems. Previous studies have shown the toxic effects of benfluralin on non-target organisms; however, its developmental toxicity in vertebrates has not yet been reported. This study demonstrated the developmental toxicity of benfluralin and its mechanism of action, using zebrafish as an aquatic vertebrate model. Benfluralin induces morphological and physiological alterations in body length, yolk sac, and heart edema. We also demonstrated a reactive oxygen species (ROS) increase of approximately 325.53 % compared with the control group after 20 μM benfluralin-treatment. In addition, the malformation of the heart and vascular structures was identified using transgenic flk1:eGFP zebrafish models at 20 μM concentration benfluralin exposure. Moreover, benfluralin induced small livers, approximately 59.81 % of normal liver size, via abnormal development of the liver as observed in the transgenic L-fabp:dsRed zebrafish. Benfluralin also inhibits normal growth via abnormal expression of cell cycle regulatory genes and increases oxidative stress, inflammation, and apoptosis. Collectively, we elucidated the mechanisms associated with benfluralin toxicity, which lead to various abnormalities and developmental toxicities in zebrafish. Therefore, this study provides information on the parameters used to assess developmental toxicity in other aquatic organisms, such as herbicides, pesticides, and environmental contaminants.

Synthesis and bioactivity evaluation of novel acetamide-derived compounds bearing a methylsulfonyl unit

This study reports the synthesis and structural characterization of a series of novel acetamide-derived compounds containing a methylsulfonyl unit. The in vitro antibacterial activity results demonstrated that compound 2-((4-fluorophenyl)sulfonyl)-N-(4-(methylsulfonyl)phenyl)acetamide (6b) exhibited superior antibacterial activities against Xanthomonas oryzae pv. oryzicolaby (Xoc), Xanthomonas oryzae pv. oryzae (Xoo), and Xanthomonas axonopodis pv. citri (Xac) at 100 μg/mL concentration, with the inhibition rates of 86.24%, 81.59%, and 75.65%, respectively, compared to Bismerthiazol and Thiodiazole copper. Meanwhile, the in vitro antifungal activities of all target compounds were found to be lower against Mucor fragilis (M. fragilis), Mucor bainieri (M. bainieri), and Trichoderma atroviride (T. atroviride) at 50 μg/mL concentration, with the inhibition rates of 0–45.21%, 0–30.24%, and 0–12.65%, respectively, compared to Carbendazim. Moreover, bioassay results of the target compounds against prostate cancer (PC3), hepatocellular carcinoma (HepG2), and leukemia (K562) cells at 10 μM concentration demonstrated that, compared to 5-fluorouracil, compound 2-((4-fluorophenyl)thio)-N-(4-(methylsulfonyl)phenyl)acetamide (4c) illustrated obvious antitumor activity (56.87%) against K562. This work first highlights the effect of the modification of the acetamide-derived compounds bearing a methylsulfonyl unit on the bioactivity evaluation.

Novel 5-aminopyrazoles endowed with anti-angiogenetic properties: Design, synthesis and biological evaluation

The promising anti-angiogenetic properties of previously synthesized pyrazolyl ureas provided the rationale for the synthesis of novel 5-aminopyrazoles 2–5, differently decorated on the pyrazole nucleus. All the derivatives were tested by MTT assays and proved to be non-cytotoxic against eight different tumor cell lines and normal fibroblasts. An EdU proliferation assay was carried out on human foreskin fibroblasts and VEGF stimulated human umbilical vein endothelial cells which confirmed the absence of cytotoxicity of the compounds on human cells up to 20 μM concentration. To evaluate the influence of the newly synthesized pyrazoles on MAPK and PI3K signaling pathways, the phosphorylation of ERK1/2 and Akt was analyzed by Western blots from HFF and HUVEC cell lysates stimulated with growth factors in the presence or absence of the compounds. Pyrazoles 3b and 3c showed a significant inhibition of Akt phosphorylation in both tested cell lines with lower phosphorylation levels than the reference compound GeGe-3 in HUVEC. Furthermore, derivatives 2 and 3 appeared to strongly affect the migration of HFF cells in a wound healing assay, confirming their potential ability to interfere with the angiogenesis process. The new pyrazole library extends the structure-activity relationships of the previously isolated compounds and highlights the attractiveness of this chemical class for pathological cell migration and angiogenesis.

Microplastics and Tire Wear Particles in Urban Stormwater: Abundance, Characteristics, and Potential Mitigation Strategies.

Stormwater has been identified as a pathway for microplastics (MPs), including tire wear particles (TWPs), into aquatic habitats. Our knowledge of the abundance of MPs in urban stormwater and potential strategies to control MPs in stormwater is still limited. In this study, stormwater samples were collected from microlitter capture devices (inlet and outlet) during rain events. Sediment samples were collected from the material captured in the device and from the inlet and outlet of a constructed stormwater wetland. MP (>25 μm) concentration in stormwater varied across different locations ranging from 3.8 to 59 MPs/L in raw and 1.8 to 32 MPs/L in treated stormwater, demonstrating a decrease after passage through the device (35-88% removal). TWPs comprised ∼95% of all particles, followed by polypropylene (PP) and poly(ethylene terephthalate) (PET). The concentration of TWPs ranged from 2.5 to 58 TWPs/L and 1450 to 4740 TWPs/kg in stormwater and sediment, respectively. A higher abundance of MPs was found in the sediment at the inlet of the constructed wetland compared to the outlet, indicating a potential role of wetlands in removing MPs from stormwater. These findings suggest that both constructed wetlands and microlitter capture devices can mitigate the transport of MPs from stormwater to the receiving waterways.

Enhancing the reliability of particulate matter sensing by multivariate Tobit model using weather and air quality data

Low-cost particulate matter (PM) sensors have been widely used following recent sensor-technology advancements; however, inherent limitations of low-cost monitors (LCMs), which operate based on light scattering without an air-conditioning function, still restrict their applicability. We propose a regional calibration of LCMs using a multivariate Tobit model with historical weather and air quality data to improve the accuracy of ambient air monitoring, which is highly dependent on meteorological conditions, local climate, and regional PM properties. Weather observations and PM2.5 (fine inhalable particles with diameters ≤ 2.5 μm) concentrations from two regions in Korea, Incheon and Jeju, and one in Singapore were used as training data to build a visibility-based calibration model. To validate the model, field measurements were conducted by an LCM in Jeju and Singapore, where R2 and the error after applying the model in Jeju improved (from 0.85 to 0.88) and reduced by 44% (from 8.4 to 4.7 μg m−3), respectively. The results demonstrated that regional calibration involving air temperature, relative humidity, and other local climate parameters can efficiently correct the bias of the sensor. Our findings suggest that the proposed post-processing using the Tobit model with regional weather and air quality data enhances the applicability of LCMs.

Influence of natural and anthropogenic aerosols on cloud base droplet size distributions in clouds over the South China Sea and West Pacific

Abstract. Cumulus clouds are common over maritime regions. They are important regulators of the global radiative energy budget and global hydrologic cycle, as well as a key contributor to the uncertainty in anthropogenic climate change projections due to uncertainty in aerosol–cloud interactions. These interactions are regionally specific owing to their strong influences on aerosol sources and meteorology. Here, our analysis focuses on the statistical properties of marine boundary layer (MBL) aerosol chemistry and the relationships of MBL aerosol to cumulus cloud properties just above cloud base as sampled in 2019 during the NASA Cloud, Aerosol and Monsoon Processes Philippines Experiment (CAMP2Ex). The aerosol and clouds were sampled by instruments on the NASA P-3 aircraft over three distinct maritime regions around the Philippines: the West Pacific, the South China Sea, and the Sulu Sea. Our analysis shows three primary sources influenced the aerosol chemical composition: clean marine (ocean source), industrial (Southeast Asia, Manila, and cargo and tanker ship emissions), and biomass burning (Borneo and Indonesia). The clean marine aerosol chemical composition had low values of all sampled chemical signatures, specifically median values of 2.2 µg m−3 of organics (ORG), 2.3 µg m−3 of SO4, 0.3 µg m−3 of NO3, 1.4 µg m−3 of NH4, 0.04 µg m−3 of Cl, and 0.0074 µg m−3 of refractory black carbon (BC). Chemical signatures of the other two aerosol source regions were industrial, with elevated SO4 having a median value of 6.1 µg m−3, and biomass burning, with elevated median concentrations of ORG 21.2 µg m−3 and BC 0.1351 µg m−3. Based on chemical signatures, the industrial component was primarily from ship emissions, which were sampled within 60 km of ships and within projected ship plumes. Normalized cloud droplet size distributions in clouds sampled near the MBL passes of the P-3 showed that clouds impacted by industrial and biomass burning contained higher concentrations of cloud droplets, by as much as 1.5 orders of magnitude for diameters &lt; 13 µm compared to clean marine clouds, while at size ranges between 13.0–34.5 µm the median concentrations of cloud droplets in all aerosol categories were nearly an order of magnitude less than the clean marine category. In the droplet size bins centered at diameters &gt; 34.5 µm concentrations were equal to, or slightly exceeded, the concentrations of the clean marine clouds. These analyses show that anthropogenic aerosols generated from industrial and biomass burning sources significantly influenced cloud base microphysical structure in the Philippine region enhancing the small droplet concentration and reducing the concentration of mid-sized droplets.

Pyrrolidinedione-thiazolidinone hybrid molecules with potent cytotoxic effect in squamous cell carcinoma SCC-15 cells

The hybrid heterocyclic molecules are perspective materials in the development of anticancer drugs. Here, the pyrrolidinedione-thiazolidinone hybrid molecules were designed as potent anticancer agents. This study aimed to investigate the cytotoxic effect of three derivatives 1-(4-hydroxyphenyl)-, 1-(4-chlorophenyl)- and 1-(4-bromophenyl)-3-[5-[2-chloro-3-(4-nitrophenyl)prop-2-enylidene]-4-oxo-2-thioxothiazolidine-3-yl]pyrrolidine-2,5-diones (Les-6287, Les-6294, and Les-6328, respectively), their effect on the production of the reactive oxygen species (ROS), apoptosis induction, and expression of genes - PPARγ, AHR, and NRFL2 – whose products are important in metabolism in human tongue squamous cell carcinoma cells of SCC-15 line. The results of resazurin reduction and lactate dehydrogenase (LDH) release assays proved the toxicity of the tested derivatives for the SCC-15 cells. Les-6287, Les-6294, and Les-6328 inhibited the viability of SCC-15 cells with the half-maximal effective concentration (EC50) in the range of 10.18–32.75 µM at 24 and 48 h treatment. These derivatives reduced the metabolism of SCC-15 cells with the half-maximal inhibitory concentration (IC50) of 6.72–39.85 µM at 24 and 48 h treatment. Les-6287, Les-6294, and Les-6328 reduced the metabolism of normal human keratinocytes of HaCaT line murine fibroblasts of Balb/c 3T3 line to a lesser extent. The compounds used in a range from 50 to 100 µM concentrations decreased ROS production in the SCC-15 cells. The derivatives Les-6287 and Les-6328 decreased the level of expression of mRNA of PPARγ, AHR, and NRFL2 genes in these cells at PPARγ siRNA knockdown and without it. Thus, the anticancer effect of studied hybrid pyrrolidinedione-thiazolidinones in the SCC-15 carcinoma cells is accompanied by a reduction of their metabolic activity and ROS level, and increase in caspase 3 activity. However, these changes are not the result of direct interaction of Les-6287, Les-6294, and Les-6328 with the PPARγ molecule.

Suppression of human lysozyme aggregation by a novel copper-based complex of 3,4-dimethoxycinnamic acid

In this work, a new Cu(II)-based complex as a chemotherapeutic drug agent, formulated as[Cu(DCA)4(H2O)2]⋅4H2O⋅4MeOH, (DCA = 3,4-dimethoxycinnamic acid), namely 1 was successfully synthesized utilizing DCA as a ligand to arrest fibrillation in Human lysozyme. The 1 was thoroughly characterized by single crystal X-ray diffraction (SC-XRD), spectroscopic (UV-Vis and FTIR) techniques, PXRD, and TGA analysis. Its crystal structure reveals a paddle wheel network around central copper metal ions. The Cu(II) metal ions exhibit a distorted square pyramidal configuration. The fluorescence titration studies showed moderate binding interaction of 1 with HuL with Ka of 6.3x105 M−1 at pH-2, 25 °C due to its interaction withAsp53, Tyr63, Val110, and Ala111 as shown by docking and simulation studies. 1suppresses the HuL fibrillation in a concentration-dependent manner, as demonstrated by ThT assay. At 200 µM concentration, it leads to the formation of smaller species of the protein in comparison to the control sample, as suggested by Light Scattering studies. The species formed are less hydrophobic and retain their native α-helix structure compared to the control samples, which are hydrophobic and form β-sheet rich amyloids as shown by ANS hydrophobicity assay and CD spectroscopy, respectively. Furthermore, morphological analysis of the species by AFM has demonstrated that, unlike mature amyloid fibrils in the control sample, HuL forms small-size aggregates in the presence of 1 under similar fibrillation conditions. It can be concluded that 1 effectively suppresses HuL fibrillation due to moderate binding to the protein. Communicated by Ramaswamy H. Sarma

Autophagy perturbation upon acute pyrethroid treatment impacts adipogenic commitment of mesenchymal stem cells

Environmental chemical exposure can cause dysregulation in adipogenesis that can result in metabolic syndrome, which includes insulin resistance, type 2 diabetes, cardiovascular disease, as well as excessive body weight. The role of autophagy in adipocyte differentiation is debatable since both positive and negative effects have been reported. Type-I and type-II synthetic pyrethroids α-cypermethrin (CPM) and permethrin (PER), respectively, are reported to increase adipogenesis in vitro and in vivo. However, it is not known how these pyrethroids affect mesenchymal stem cells (MSCs). Thus, this study focused on evaluating the effect of pyrethroids (CPM and PER) pre-treatment (24 h) on MSC commitment and the regulatory role of autophagy in adipogenic lineage commitment. The formation of adipocytes was observed through nile red staining, perilipin expression by immunoflourescence, and adipogenic markers PPARγ, C/EBPα, and FABP4 by western blotting. It was found that the adipogenic differentiation ability of MSCs was significantly increased upon CPM or PER pre-treatment at 100 μM concentration as evident by lipid accumulation and enhanced expression of adipogenic markers. To assess the involvement of autophagy, the expression of p62 and LC3II were evaluated following pre-treatment. Immunoblotting results revealed an increased expression of p62 and LC3II in CPM or PER pretreated MSCs suggesting CPM and PER mediated inhibition of autophagy at 24 h. Further, an increase was observed in adipogenesis upon CPM or PER pre-treatment in combination with chloroquine, while use of rapamycin during pre-treatment abrogated the effect of CPM and PER. Thus, this study concludes that CPM or PER pre-treatment increases the adipogenic differentiation of MSCs. Since chloroquine also demonstrated similar adipogenic response, it further highlights that 24 h pre-treatment with autophagy modulators to inhibit basal autophagy primes MSCs towards adipogenic lineage.

Sulfate-rich spring seeps and seasonal formation of terraced, crystalline mirabilite mounds along the shores of Great Salt Lake, Utah: Hydrologic and chemical expression during declining lake elevation

As Great Salt Lake (GSL) elevation declined below ∼1278.6 m (4195 ft) beginning in the fall of 2019, crystalline mounds formed on the southeastern shore, near Great Salt Lake State Park, Utah. Soon after, several more mound complexes were discovered on nearby Antelope Island. Recent historic low lake levels permitted sulfate-saturated spring brines to emerge on the exposed shoreline. When the spring waters encountered cold winter air, sodium sulfate precipitated in the form of the mineral mirabilite (Na2SO4·10H2O). Mirabilite-saturated spring waters pool in shallow sediment-collapse depressions or, with sufficient hydraulic head, precipitate subaerial crystalline spring terraced mounds. The mounds are composed of clear, bladed, and tabular mirabilite crystals (1 to 10 cm in length; 1 to 5 cm in width) that form mini pools and rimstone dams due to cascading spring water flow. The waters have elavated μM concentrations of sulfide, an oxidation-reduction potential (ORP) of −250 mV, pH of 7, densities of 1.103–1.1169 g/cm3, and temperatures of 10°–15 °C, as compared to nearby lake waters that had undetectable sulfide, an ORP of 12 mV, pH of 8.2, density of 1.0872 g/cm3, and temperatures of 0°–3 °C. Green algae and probable sulfide oxidizing bacteria thrive in the spring waters and some are occluded within the mirabilite matrix and fluid inclusions. Sulfur δ34S and oxygen δ18O isotopes of brine and mirabilite sulfate are enriched +18.8–20.41‰ and + 12.2–14.5‰, respectively, indicating groundwater and mirabilite are in equilibrium for oxygen. Brine sulfide is heavily fractionated (δ 34S of ∼ − 30 ‰) relative to sulfate, indicating active sulfate reduction in subsurface aquifers that feed the spring seeps and which is consistent with higher HS− concentrations relative to adjacent lake waters. The recognition of such biosignatures retained in mirabilite provides further insights for the preservation potential of biological material in extraterrestrial sulfate minerals.Today, Gilbert Bay (south arm of GSL) is undersaturated with respect to mirabilite. However, pre-earthen causeway historical reports (prior to 1959) indicate mirabilite precipitated in the south arm during the winter and through wave action accumulated along the leeward southeastern oolitic shore (similar to current processes occurring in the hypersaline Gunnison Bay, the north arm of GSL). Upon solar irradiance in the spring, the shoreline mirabilite dissolved and reprecipitated in the shallow subsurface (<1 m) as an ooid cement. We hypothesize that this subsurface layer of mirabilite is the source for active lake margin sodium sulfate-saturated spring brines. The emergence and surface exposure of the brines and associated chemical sediments are an additional contributing solute source that may increase GSL salinity during lake level decline more than evaporation alone.

Effects of melatonin foliar application on hot pepper growth and stress tolerance

PurposeDrought and waterlogging are one of the most severe abiotic stresses towards plant growth and development. The experiment was implemented in the Isfahan University of Technology's research greenhouse to examine the effect of melatonin foliar spraying in hot pepper plants under waterlogging and drought stresses. MethodsThis factorial experiment was performed as a complete randomized design (CRD) in triplicate. The treatments included controls, waterlogging, drought stress, and melatonin foliar spraying at 0 and 500 µM concentrations. ResultsThe results demonstrated that the foliar melatonin spraying was more advantageous under drought and waterlogging stress conditions via increasing ammonium content, NR (nitrate reductase activity), chlorophyll content, proline, protein, transpiration, shoot length, root water content, amino acid alterations, and PIP gene expression. ConclusionsMelatonin influenced the hot pepper's growth by affecting enzyme activity NR hormonal changes (ABA), amino acid changes, and PIP gene expression.

Insights into thiram fungicide: A comparative study of solution and solid phases through Raman, DCDR, and SERS measurements and DFT simulations

AbstractThiram, a widely known fungicide, has evinced a negative impact on the environment and human health that led to the banning of its use in plant protection products on all commodities in the European Union. Therefore, there is a need for monitoring possible illicit use, which requires sensitive and accurate detection methods. In this study, the behavior of thiram in different phases was investigated in order to better understand its properties. Raman spectra were obtained from thiram in the solid state as a crystalline powder, dissolved in different solvents, and from dried deposits using the drop coating deposition Raman (DCDR) method. The analyses of acquired Raman spectra revealed evidence of hydrogen bonding between thiram and chloroform in the solution phase. The reliability and sensitivity of DCDR measurements were also highlighted, with high‐quality spectra obtained from a dried pattern from droplet at a 40 μM concentration. Moreover, the study identified the most abundant conformer of intact (undegraded) thiram molecule and enabled reliable band assignment and vibrational analysis based on DFT simulations, providing a better understanding of the compound's properties. The results also suggested that thiram can be coordinated to silver in nanostructured SERS active substrates not only in the degraded form but also in the undegraded form. Overall, this study provides valuable insights into thiram's behavior and can be helpful in the further development of efficient and effective methods for its detection and analysis.

Efficiency of some Jasmonic Acid Concentrations on Mortality of Two-Spotted Spider Mite Tetranychus urticae Koch on Eggplant

The efficiency of Jasmonic acid concentrations 0, 5, 50 and 100μM was examined mortality of adult female and nymphs of the two-spotted spider mite Tetranychus urticae fed on leaves of over four time intervals of 12, 24, 36 and 48 hours. The results showed the significant effect of the treatments on the corrected mortality ratios for both stages, as well as for the interaction of the treatments of variety, concentration and time. In general, the results indicated the increase of mortality with the increase of the time period. It was observed that the concentration of 100μM during the time periods and all varieties and the concentration of 50μM after the time periods 24, 36 and 48 hours on the Barcelona variety and the concentration of 5μM during the last two time periods on the Bowie variety had the highest corrected mortality ratio, which was 100%, while the lowest mortality ratio was 0% at the concentration 5μM with 12 hours after treatment on adult females fed on the leaves of Jawaher variety. Regarding the effect of treatment on nymphal stage, In general, the results of the triple interaction between treatments indicated that the concentration of 5 μM had a synergistic effect along with Bowie variety and at all time periods in obtaining of the highest mortality ratio of individuals of nymphal stage (100%) and with significant differences from the other treatment while the same concentration after 12 hours of treatment achieved lowest mortality ratio of the nymph fed on the leaves of the variety Jawaher which amounted 0%.

Preclinical Pharmacokinetics and CYP Modulation Activity of Chebulinic Acid: A Potent Molecule Against Metabolic Disease.

Chebulinic acid (CA) is an active constituent of Terminalia chebula fruits with therapeutic potential against multiple metabolic diseases, including dementia, benign prostate hyperplasia, and osteoporosis. The present work intends to explore the preclinical pharmacokinetics, including the absolute bioavailability of CA and its influence on the gene expression of cytochrome P450 enzymes in the liver. Quantifying CA and probe drugs in vitro samples and preclinical serum samples of male SD rats were performed using LC-MS/MS. The influence of CA on the hepatic CYPs and their gene expression was analyzed in rat liver by quantitative real-time polymerase chain reaction. The plasma protein binding was found to be 84.81 ± 7.70 and 96.34 ± 3.12, blood-to-plasma ratio of 0.62 ± 0.16 and 0.80 ± 0.23 at 1 μM and 10 μM concentrations, respectively. Again, the absolute oral bioavailability of CA at 100 mg/kg was found to be 37.56 ± 7.3%. The in-vivo pharmacokinetic profile of probe drugs revealed CA to have significant inducing effects on CYP1A2, 2C11, 2D2, and 2E1 after 14 days, which correlates to both in-vitro rat microsomal data and gene expression results. Altogether, pharmacokinetic parameters reveal CA to have an affinity to distribute across different extravascular tissues and induce rat liver CYP enzymes.