Computational and experimental investigations of a novel aptamer targeting oxidized low-density lipoprotein.

Oxidized low-density lipoprotein (oxLDL) induces the formation of atherosclerotic plaques. Apolipoprotein B100 (apoB100) is a crucial protein component in low-density lipoprotein (LDL), which includes oxLDL. The oxidation of amino acids and subsequent alterations in their structure generate oxLDL, which is a significant biomarker for the initial phases of coronary artery disease. This study employed molecular docking and molecular dynamics utilizing the MM/GBSA method to identify aptamers with a strong affinity for oxidized apoB100. Molecular docking and molecular dynamics were performed on two sequences of the aptamer candidates (aptamer no.11 (AP11: 5'-CTTCGATGTAGTTTTTGTATGGGGTGCCCTGGTTCCTGCA-3') and aptamer no.26 (AP26: 5'-GCGAACTCGCGAATCCAGAACGGGCTCGGTCCCGGGTCGA-3')), yielding respective binding free energies of -149.08 kcal/mol and -139.86 kcal/mol. Interaction modeling of the simulation revealed a strong hydrogen bond between the AP11-oxidized apoB100 complexes. In an aptamer-based gold nanoparticle (AuNP) aggregation assay, AP11 exhibits a color shift from red to purple with the highest absorbance ratio, and shows strong binding affinity to oxLDL, correlating with the simulation model results. AP11 demonstrated the potential for application as a novel recognition element in diagnostic methodologies and may also contribute to future advancements in preventive therapies for coronary artery disease.

Comparative effects of heat stress on photosynthesis and oxidative stress in Halophila ovalis and Thalassia hemprichii under different light conditions.

This study investigated the physiological responses of two tropical seagrass species, Halophila ovalis and Thalassia hemprichii, to heat stress under varying light conditions in a controlled 5-day experiment. The experimental design included four treatments: control, saturating light, heat stress under sub-saturating light, and heat stress under saturating light (combined stress). We assessed various parameters, including chlorophyll fluorescence, levels of reactive oxygen species (ROS), antioxidant enzyme activities, and growth rates. In H. ovalis, heat stress resulted in a significant reduction in the maximum quantum yield of photosystem II (Fv/Fm) regardless of the light condition. However, the effects of heat stress on the effective quantum yield of photosystem II (ɸPSII) were more pronounced under saturating light conditions. In T. hemprichii, saturating irradiance exacerbated the heat stress effects on Fv/Fm and ɸPSII, although the overall photoinhibition was less severe than in H. ovalis. Heat stress led to ROS accumulation in H. ovalis and reduced the activity of superoxide dismutase (SOD) and ascorbate peroxidase in the sub-saturating light condition. Conversely, T. hemprichii exhibited elevated SOD activity under saturating light. Heat stress suppressed the growth of both seagrass species, regardless of the light environment. The Biomarker Response Index indicated that H. ovalis displayed severe effects in the heat stress treatment under both light conditions, while T. hemprichii exhibited moderate effects in sub-saturating light and major effects in saturating light conditions. However, the Effect Addition Index revealed an antagonistic interaction between heat stress and high light in both seagrass species. This study underscores the intricate responses of seagrasses, emphasizing the importance of considering both local and global stressors when assessing their vulnerability.

A simple aptamer/gold nanoparticle aggregation-based colorimetric assay for oxidized low-density lipoprotein determination.

Oxidized low-density lipoprotein (oxLDL) is the leading cause of atherosclerosis and cardiovascular diseases. Here, we created a simple colorimetric assay for sensitive and specific determination of oxLDL using a selective aptamer coupled with salt-induced gold nanoparticle (AuNP) aggregation. The aptamer was chosen by Systematic Evolution of Ligands by Exponential Enrichment to obtain a novel selective sequence towards oxLDL (as 5'-CCATCACGGGGCAGGCGGACAAGGGGTAAGGGCCACATCA-3'). Mixing a 5 µM aptamer solution with an aliquot of a sample containing oxLDL followed by adding AuNP solution (OD = 1) and 80 mmol L-1 NaCl achieved rapid results within 19 min: linear response to oxLDL from 0.002 to 0.5 µmol L-1 with high selectivity, a recovery accuracy of 100-111% at the 95% confidence interval, and within-run and between-run precision of 1-6% and 1-5% coefficient variations, respectively. Artificial serum diluted at least 1:8 with distilled water, analyzed by the aptamer-based colorimetric assay, showed excellent correlation with conventional thiobarbituric acid reactive substances (TBARS) (R2 = 0.9792) as a rapid colorimetric method without the need for sample preparation other than dilution.

The complete mitogenome of the Thai soldier crab Mictyris thailandensis Davie, Wisespongpand & Shih, 2013 (Crustacea: Decapoda: Mictyridae).

Mictyris thailandensis has been described recently in the family Mictyridae which is found only in the Andaman Sea, west coast of Thailand. In this study, we performed shotgun genome sequencing of a male M. thailandensis using a paired-end (150 bp) sequencing chemistry on MGISEQ-2000RS and report the complete mitochondrial genome of M. thailandensis (15,557 bp). A total of 37 genes have been annotated: 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and a control region. Comparative phylogenetic analysis with 29 crustaceans based on 13 conserved genes demonstrated that M. thailandensis is closely related to other soldier crabs in the family Mictyridae. The mitogenome of M. thailandensis presented here provides useful genetic information to help understand the evolutionary relationships among the Mictyridae family members.

Copper and zinc differentially affect root glutathione accumulation and phytochelatin synthase gene expression of Rhizophora mucronata seedlings: Implications for mechanisms underlying trace metal tolerance.

Mangroves are susceptible to contamination due to their proximity to shores and human activities. Exposure to excessive trace metals can disturb their physiological functions and may eventually lead to death. Rhizophora mucronata is a common species growing in the mangrove forests of Thailand. Previous studies have shown that seedlings of R. mucronata are tolerant of trace metal and that they accumulate a large metal content in their root tissue. However, knowledge of their tolerance mechanisms is still lacking. To elicit the role of metal detoxification and sequestration by phytochelatins (PC) in the roots of R. mucronata seedlings, the impacts of Cu and Zn exposure were assessed on 1) physiological characteristics 2) the concentration of glutathione (GSH), a precursor of PC and 3) the level of the transcripts encoding phytochelatin synthase (PCS), the key enzyme for PC biosynthesis. Seedlings of R. mucronata were exposed to Cu and Zn in a hydroponic experiment (200 mg Cu or Zn/L in 1/4× Hoagland solution containing 8‰ NaCl, single addition). We found that both trace metals were largely accumulated in the roots. Only Cu-treated seedlings showed a decrease in the photosynthetic efficiency, in line with observed toxicity symptoms (i.e. bent stems and slight wilting of leaves). Metal accumulation, however, did not induce oxidative stress in the roots as indicated by similar level of total reactive species and lipid peroxidation across treatments. The GSH content in the roots exposed to Cu was significantly reduced while no change was observed in Zn-exposed roots. Coordinated semi-quantitative PCR and RT-qPCR revealed pcs down-regulation in Cu-treated roots, whereas Zn-treated roots showed a down-regulation on day 1 and a subsequent recovery on day 5. Failure of detoxification and sequestration of excess Cu due to GSH limitation and down-regulation of pcs may lead to the phytotoxic effects observed in Cu-treated plants. Our results suggest that both GSH and PC play an important role in trace metal tolerance in R. mucronata seedlings.

Treatment with Pluchea indica (L.) Less. leaf ethanol extract alleviates liver injury in multiple low-dose streptozotocin-induced diabetic BALB/c mice.

Hyperglycemia-induced oxidative stress and inflammation are hallmarks of liver damage in diabetes mellitus. Accumulating evidence has demonstrated that Pluchea indica leaf ethanol extract (PILE) possesses strong antioxidant and anti-inflammatory properties. However, studies of its effects on liver damage in streptozotocin (STZ)-induced diabetic animals remain insufficient. To the best of our knowledge, the present study was the first to illustrate that PILE mitigated liver injury in STZ animals. Mice were first pretreated with PILE at either 50 mg/kg (PILE 50) or 100 mg/kg (PILE 100) 2 weeks prior to the induction of hyperglycemia by multiple low doses of STZ. The mice were then fed with PILE 50 or PILE 100 for 4 or 8 weeks, following which liver weight, pathological changes, oxidative stress parameters, inflammation-related markers and caspase-mediated apoptosis were measured at each time point. Untreated STZ mice exhibited abnormal increases in liver weight and severe pathological changes. However, PILE 100 reduced the severity of the STZ-induced diabetic phenotype at both time points. A significant decrease in the levels of superoxide dismutase and catalase, in addition to an increase in malondialdehyde, were observed in the livers of untreated STZ mice, all of which were significantly reversed by treatment with PILE 100 for 8 weeks. Western blot analysis revealed reduced levels of liver inflammatory markers, including interleukin-6, tumor necrosis factor-α, NF-κB p65, transforming growth factor-ß1 and protein kinase C following PILE 100 treatment. Additionally, changes in the levels of apoptotic markers indicated that PILE 100 significantly attenuated caspase-9 and -3 expression, whilst preserving that of the Bcl-2 protein. In conclusion, the present study revealed that PILE alleviates hyperglycemia-induced liver injury by normalizing the various mediators of oxidative stress, inflammation and apoptosis.

Ethanolic extracts of Pluchea indica (L.) leaf pretreatment attenuates cytokine-induced ß-cell apoptosis in multiple low-dose streptozotocin-induced diabetic mice.

Loss of ß-cell mass and function is a fundamental feature of pathogenesis for type 1 and type 2 diabetes. Increasing evidence indicates that apoptosis is one of the main mechanisms of ß-cell death in both types. Ethanolic extracts of Pluchea indica leaf (PILE) have been reported to possess blood glucose lowering actions in vivo. Nevertheless, further study is required to determine the underlying mechanisms. In this report, we have investigated the preventive effects of PILE on multiple low doses of streptozotocin (MLDS)-induced ß-cell apoptosis. Mice were pre-treated with PILE at 50 mg/kg (PILE 50) or 100 mg/kg (PILE 100) for 2 weeks before streptozotocin (STZ) stimulation, and the treatment continued for 4 or 8 weeks. Results revealed that PILE 100 mice exhibited improved blood biochemistry, maintained a higher body weight, had decreased hyperglycemia, and restored islet architectures compared to non-treated STZ mice. Significantly, PILE 100 decreased levels of inflammatory response markers interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interlukin1-ß (IL-1ß), concomitant with the inhibition of caspase-3, caspase-8, capsepase-9, phosphorylation of signal transducer and activator of transcription 1 (pSTAT1), nuclear factor-κBp65 (NF-κBp65), and inducible nitric oxide synthase (iNOS). Additionally, survival and proliferative ability of ß-cells was mediated by up-regulated Bcl-2 and Ki67, respectively. These results provide strong evidence that pretreatment with PILE 100 effectively attenuated STZ-induced diabetes-related symptoms and these effects could be associated with the inhibition of cytokine-induced ß-cell apoptosis.

A purple non-sulfur bacterium producing polyhydroxybutyrate and the conserved region of pha synthase gene / Bactéria púrpura não sulfurosa produtora de polihidroxibutirato e a região conservada do gene pha sintase

This study aimed to screen purple non-sulfur bacteria capable of accumulating granules or polyhydroxybutyrate (PHB) inside the cells, identify the potent strain, assay the enzyme or PHA synthase, and compare the PHB synthase gene with that of related strains. A total of 58 strains of purple non-sulfur bacteria were isolated from 108 samples of chicken feces in the chicken-egg farm of the Department of Animal Science, Faculty of Natural Resources at Prince of Songkla University, Hat Yai, Thailand. After cultivating the bacteria in glutamate malate (GM) medium without added glutamic acid under light (3,000 Lux) at 35oC for 5 days, the intracellular biopolymer granules of the bacteria were observed by using a Confocal Laser Scanning Microscope (CLSM) with excitation and emission wavelength of 530 and 605 nm, respectively. Gas chromatography (GC) was carried out for quantitative analysis of PHB. There were five strains, CH12, CH52, CH72, CH90 and CH92, showed biopolymer granules under CLSM, and accumulated PHB 5, 1.7, 1.5, 1.4 and 1.8% (w w-1) of the cell dry weight (CDW), respectively. The 16S rDNA sequence analysis of CH12 strain showed a high homology of 100% correlation to that of Rhodopseudomonas palustris strain NCIB8288. Regarding the taxonomic characteristics and 16S rDNA sequence analysis, CH12 strain was identified as Rps. palustris NCIB8288. The PHA synthase activity of the crude extract from CH12 strain was 25 units/mL. The conserved regions could be aligned and selected among 5 strains of Rhodopseudomonas palustris (strains BisA53, TIE-1, CGA009, HaA2 and BisB18). The purified PCR product was obtained for further studies.

Este estudo teve como objetivo rastrear bactérias púrpuras não sulfurosas capazes de acumular grânulos ou polihidroxibutirato (PHB) dentro das células, identificar a estirpe potente, ensaiar a enzima ou PHA sintaxe, e comparar com o gene PHB sintase com aquele de estirpes relacionadas. Um total de 58 estirpes de bactérias púrpuras não sulfurosas foram isoladas a partir de 108 amostras de fezes de galinhas na granja produtora de ovos do Departamento de Ciência Animal, Faculdade de Recursos Naturais da Universidade Prince of Songkla, Hat Yai, Tailândia. Depois de cultivar as bactérias em um substrato de glutamato/malato (GM), sem ácido glutâmico adicionado, sob luz (3000 lux) a 35 ºC durante 5 dias, os grânulos de biopolímeros intracelulares das bactérias foram observados utilizando um microscópio confocal (do inglês Confocal Laser Scanning Microscope - CLSM) com comprimentos de onda de excitação e emissão de 530 e 605 nm, respectivamente. A cromatografia gasosa (do inglês Gas chromatography - GC) foi realizada para uma análise quantitativa de PHB. Havia 5 estirpes, CH12, CH52, CH72, CH90 e CH92, que mostraram grânulos biopoliméricos quando submetidos ao CLSM, e PHB-5 acumulado de 1.7, 1.5, 1.4 and 1.8% (w w-1) do peso celular seco (do inglês cell dry weight - CDW), respectivamente. A análise da sequência do rDNA 16S da estirpe CH12 demonstrou uma alta correlação de homologia de 100% para aquela da estirpe NCIB8288 da Rhodopseudomonas palustris. Em relação às características taxonômicas e da análise da sequência do rDNA 16S, a estirpe CH12 foi identificada como Rps. palustris NCIB8288. A atividade da PHA sintase do extrato bruto da estirpe CH12 foi de 25 unidades/mL. As regiões conservadas puderam ser alinhadas e selecionadas entre 5 estirpes de Rhodopseudomonas palustris (BisA53, TIE-1, CGA009, HaA2 e BisB18). O produto purificado da reação em cadeia da polimerase - PCR foi obtido para estudos futuros.