Total Fluorescence Research Articles

How do metals interact with periphytic biofilms?

Extracellular matrix of periphyton has complex structural and chemical composition regulating metal transfer within biofilms with consequences for metal transfer to aquatic food webs. We investigated which metal species were retained in the loosely (LB) and the tightly bound (TB) fractions of the periphyton matrix from three pristine lakes at different growth stages. We measured the fluorescent dissolved organic matter (FDOM) composition with parallel factor analysis (PARAFAC) and the co-occurrence of essential and non-essential metals with FDOM in the two matrix fractions. The LB and TB fractions of periphyton had distinct fluorescence composition from the water column. The PARAFAC model identified five components, including two (C2 and C4) appearing to be of periphytic origin. The humic-like C2 was almost exclusive to periphyton and the tryptophan-like C4, associated to biofilm phototrophy, represented up to 47.0 ± 7.3 % of total fluorescence in the LB fraction. Most metals had significant positive relationships with four FDOM components in the LB fraction while C2 was the only component in the TB fraction to show such significant relationships. Components in the LB fraction seemed to act as scavengers for metals, preventing them from reaching the cellular fraction, while C2 from the TB fraction was likely promoting the bioavailability of essential metals for microorganisms inside periphyton. This study highlights the contrasting roles of the extracellular matrix on metal mobility beyond the usually proposed protection mechanisms. We suggest an experimental model for the study of metal regulation processes of the periphytic extracellular polymeric substances with a focus on the components produced by microorganisms within periphyton and their distribution in the different matrix fractions.

Rapid, simultaneous and non-destructive determination of multiple adulterants in Panax notoginseng powder by front-face total synchronous fluorescence spectroscopy

The authentication of traditional herbal medicines in powder form is of great significance, as they are always of high values but vulnerable to adulteration. Based on the distinct fluorescence of protein tryptophan, phenolic acids and flavonoids, front-face synchronous fluorescence spectroscopy (FFSFS) was applied for the fast and non-invasive authentication of Panax notoginseng powder (PP) adulterated with the powder of rhizoma curcumae (CP), maize flour (MF) and whole wheat flour (WF). For either single or multiple adulterants in the range of 5–40% w/w, prediction models were built based on the combination of unfolded total synchronous fluorescence spectra and partial least square (PLS) regression, and were validated by both five-fold cross-validation and external validation. The constructed PLS2 models simultaneously predicted the contents of multiple adulterants in PP and gave suitable results, with most of the determination coefficients of prediction (Rp2) >0.9, the root mean square error of prediction (RMSEP) no >4% and residual predictive deviation (RPD) >2. The limits of detections (LODs) were 12.0, 9.1 and 7.6% for CP, MF and WF, respectively. All the relative prediction errors for simulated blind samples were between −22% and + 23%. FFSFS offers a novel alternative to the authentication of powdered herbal plants.

Carbon-based heterostructure from multi-photo-active nanobuilding blocks SrTiO3@NiFe2O4@Fe0@Ni0@CNTs with derived nanoreaction metallic clusters for enhanced solar light-driven photodegradation of harmful antibiotics

We reported on the sequential development of a high-operative photocatalyst with penta-component inorganic bulk heterojunction for improved charge trapping characteristics at particle–particle interfaces for enhanced solar light-driven photocatalytic degradation of active tetracycline antibiotic. Structural, morphological, optical, and electronic properties of the synthesized samples were investigated using a series of complementary characterization techniques, such as XRD, FE-SEM, HR-TEM, XPS, as well as hard and soft XAS in both total electron yield (TEY) and fluorescence yield (TFY). For the case of the carbon composite material, SrTiO3@NiFe2O4@Fe0@Ni0@CNTs, a reduced crystallinity when compared to the starting support material was noticed, although this translated into a significant improvement of the morphology and the photocatalytic performance. The SrTiO3@NiFe2O4@Fe0@Ni0@CNTs fibrous photocatalyst can efficiently achieve a high-to-total degradation of tetracycline antibiotic under visible light irradiation in less than two hours, following a non-linear PFO kinetic model with an apparent reaction rate of about 0.0606 min−1 and an 98% photodegradation activity. The XPS and XAS analysis demonstrated unequivocally the appearance of nanoscale zero-valent iron (Fe0) and zero-valent nickel (Ni0) on the photocatalyst surface, which facilitates the separation of photogenerated e+ and h+ pairs, and the appearance of more active sites.

Effects of drought stress on growth, physiological and biochemical parameters of two Ethiopian red pepper (Capsicum annum L.) cultivars

Drought is the most predominant abiotic factor that affects plant growth and development. The present research examined at how drought stress affected the growth, physiological, and biochemical responses of Local and Markofana red pepper (Capsicum annum L.) cultivars. Five-week-old cultivars were exposed to well-watered, low, moderate, and severe drought conditions, kept at 100, 80, 60 and 40% of field capacity, respectively. Nine-week-old cultivars were sampled to analyze the effects of the stress on different parameters of both cultivars. Compared with the control group, drought stress caused a reduction in growth, physiological and biochemical parameters; nevertheless, adverse effects of the stress were more noticeable in the Local cultivar. Severe drought stress significantly reduced shoot length in Markofana (53.71 %) compared to the control group. Significant variation was observed in relative water content in the Local cultivar (20.26 %) exposed to drought. In the Local cultivar, the total chlorophyll content and chlorophyll fluorescence declined significantly by 77.28 and 3.33 %, respectively. Therefore, the cultivar Markofana was relatively less affected by drought stress. In general, these differences in cultivar responses to drought stress may aid in developing drought tolerance genotypes that can withstand drought stress conditions with minimal yield losses.

Effects of sea-salt stress on morphological and biochemical indices of Grand Nain (Musa spp.) banana variety

Abstract Banana is cultivated abundantly in seaside regions, which are highly prone to rises in sea level or where seawater mixes with groundwater. This study aimed to investigate the effect of sea-salt stress on banana seedlings. Three different saline solutions, irrigation water, and distilled water were prepared for treatments; 15.3 dS m-1, 20.4 dS m-1, 25.7 dS m-1, 3.22 dS m-1 (irrigation water), 0.0 dS m-1 (control), respectively. Although there was no statistical difference in plant length after the treatment, plant diameter difference was observed in seedlings subjected to salt stress for 30 days. The most damaged leaves (4.8) were determined at the highest sea-salt solution (25.7 dS m-1). Compared with the root length (RL) results of control, irrigation water, and 15.3 dS m-1 treatments, the mean of RL was significantly higher in the 25.7 dS m-1 treatment (40.0 cm). Except for the 20.40 dS m-1 treatment, the other treatments had values close to each other. Contrary to the total chlorophyll (TC) and chlorophyll fluorescence (CF) values determined in control plants, the highest TC content and CF were found in irrigation water treatment with 34.8 mg g-1 Fw and 46.0, respectively. The levels of malondialdehyde (MDA) and proline were significantly higher in the 25.7 dS m-1 treatment than in the other treatments. Results demonstrated that the root length of the Grand Nain cultivar increased although the sea-salt stress had negative effects on plant morphology and biochemical content, indicating that it could adapt to short-term salt stress.

Microglial Phagocytosis/Cell Health High-Content Assay.

We report a microglial phagocytosis/cell health high-content assay that has been used to test small molecule chemical probes and support our drug discovery projects targeting microglia for Alzheimer's disease therapy. The assay measures phagocytosis and cell health (cell count and nuclear intensity) simultaneously in 384-well plates processed with an automatic liquid handler. The mix-and-read live cell imaging assay is highly reproducible with capacity to meet drug discovery research needs. Assay procedures take 4 days including plating cells, treating cells, adding pHrodo-myelin/membrane debris to cells for phagocytosis, staining cell nuclei before performing high-content imaging, and analysis. Three selected parameters are measured from cells: 1) mean total fluorescence intensity per cell of pHrodo-myelin/membrane debris in phagocytosis vesicles to quantify phagocytosis; 2) cell counts per well (measuring compound effects on proliferation and cell death); and 3) average nuclear intensity (measuring compound induced apoptosis). The assay has been used on HMC3 cells (an immortalized human microglial cell line), BV2 cells (an immortalized mouse microglial cell line), and primary microglia isolated from mouse brains. Simultaneous measurements of phagocytosis and cell health allow for the distinction of compound effects on regulation of phagocytosis from cellular stress/toxicity related changes, a distinguishing feature of the assay. The combination of cell counts and nuclear intensity as indicators of cell health is also an effective way to measure cell stress and compound cytotoxicity, which may have broad applications as simultaneous profiling measurements for other phenotypic assays. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Microglial phagocytosis/cell health high-content assay protocol Support Protocol: Procedures to isolate myelin/membrane debris from mouse brain and label with pHrodo.

A practical application of front-face synchronous fluorescence spectroscopy to rapid, simultaneous and non-destructive determination of piperine and multiple adulterants in ground black and white pepper (Piper nigrum L.)

Based on the distinct fluorescence of piperine and tryptophan, and their different profiles in pepper and several possible adulterants, front-face synchronous fluorescence spectroscopy (FFSFS) was applied for the fast and non-invasive authentication of ground black pepper adulterated with papaya seed powder and buckwheat flour, and ground white pepper adulterated with whole wheat and maize flours. For either single adulterant or dual adulterants in the range of 10–40% w/w, prediction models were constructed based on the combination of unfolded total synchronous fluorescence spectra and partial least square (PLS) regression, and were validated by both five-fold cross-validation and external validation. The built PLS2 models produced suitable results, with most of the determination coefficients of prediction (Rp2) greater than 0.8, the root mean square error of prediction (RMSEP) < 5% and residual predictive deviation (RPD) greater than 2. The limits of detection (LODs) were 11.1, 5.5, 10.6 and 12.0% for papaya seed powder, buckwheat, whole wheat and maize flours, respectively. Most relative prediction errors for simulated blind samples were within ± 30%. Besides, piperine in ground black and white pepper was also determined with acceptable PLS results.

Correlation Between Indocyanine Green Fluorescence Patterns and Grade of Differentiation of Hepatocellular Carcinoma: A Western Prospective Cohort Study.

Background. Most of the available evidence on the use of indocyanine green (ICG) fluorescence in clinical practice consists of articles published by surgeons of the Asian-Pacific area. We performed a prospective cohort study to assess the patterns of ICG fluorescence in Western hepatocellular carcinoma (HCC) counterparts.Methods. From April 2019 to January 2022, a total of 31 consecutive patients who underwent laparoscopic liver resection (LLR) for superficial HCC were enrolled in this prospective study. All patients underwent laparoscopic staging with both laparoscopic ultrasound (LUS) and ICG fluorescence imaging.Results. A total of 38 hepatocellular carcinomas (HCCs) were enrolled: 23 superficial (surfacing at the liver's Glissonian capsule), 5 exophytic, 5 shallow (<8mm from the hepatic surface) and 5 deep (>10mm from the hepatic surface). The detection rate with preoperative imaging (abdominal CT/MRI), LUS, ICG fluorescence and combined modalities (ICG and LUS) was 97.4%, 94.9%, 89.7% and 100%, respectively. The five deep seated lesions underwent ultrasound-guided laparoscopic thermal ablation. The other 33 HCCs were treated with minimally invasive liver resection. Intraoperative ultrasound patterns were registered for each single nodule resected. The ICG fluorescence pattern was classified in two types: total fluorescence (all the tumoral tissue showed strong and homogeneous fluorescence), n = 9/33 (27.3%), and non-total fluorescence (partial and rim fluorescence), n = 24/33 (72.7%). There was a statistical correlation between ICG patterns and grade of differentiation. Almost all lesions with uniform fluorescence pattern were well-differentiated HCCs (G1-G2), while partial and rim-type fluorescence pattern were more common among moderately and poorly differentiated HCCs (G3-G4) (88.9% vs 11.1%, 37.5% vs 62.5%, P = .025, respectively).Conclusions. ICG fluorescence imaging could be used to identify early the grade of HCC, ie intraoperatively, thus influencing the intraoperative treatment.

Specific fluorescent signatures for body fluid identification using fluorescence spectroscopy

Non-invasive, rapid, on-site detection and identification of body fluids is highly desired in forensic investigations. The use of fluorescence-based methods for body fluid identification, have so far remain relatively unexplored. As such, the fluorescent properties of semen, serum, urine, saliva and fingermarks over time were investigated, by means of fluorescence spectroscopy, to identify specific fluorescent signatures for body fluid identification. The samples were excited at 81 different excitation wavelengths ranging from 200 to 600 nm and for each excitation wavelength the emission was recorded between 220 and 700 nm. Subsequently, the total emitted fluorescence intensities of specific fluorescent signatures in the UV–visible range were summed and principal component analysis was performed to cluster the body fluids. Three combinations of four principal components allowed specific clustering of the body fluids, except for fingermarks. Blind testing showed that 71.4% of the unknown samples could be correctly identified. This pilot study shows that the fluorescent behavior of ageing body fluids can be used as a new non-invasive tool for body fluid identification, which can improve the current guidelines for the detection of body fluids in forensic practice and provide the robustness of methods that rely on fluorescence.

Elemental and spectral chemometric analyses of Octopus vulgaris beaks as reliable markers of capture location.

The high demand and economic relevance of cephalopods make them prone to food fraud, including related to harvest location. Therefore, there is a growing need to develop tools to unequivocally confirm their capture location. Cephalopod beaks are nonedible, making this material ideal for traceability studies as it can also be removed without a loss of commodity economic value. Within this context, common octopus (Octopus vulgaris) specimens were captured in five fishing areas along the Portuguese coast. Untargeted multi-elemental total X-ray fluorescence analysis of the octopus beaks revealed a high abundance of Ca, Cl, K, Na, S, and P, concomitant with the keratin and calcium phosphate nature of the material. We tested a suite of discrimination models on both elemental and spectral data, where the elements contributing most to discriminate capture location were typically associated with diet (As), human-related pressures (Zn, Se, and Mn), or geological features (P, S, Mn, and Zn). Among the six different chemometrics approaches used to classify individuals to their capture location according to their beaks' element concentration, classification trees attained a classification accuracy of 76.7%, whilst reducing the number of explanatory variables for sample classification and highlighting variable importance for group discrimination. However, using X-ray spectral features of the octopus beaks further improved classification accuracy, with the highest classification of 87.3% found with partial least-squares discriminant analysis. Ultimately, element and spectral analyses of nonedible structures such as octopus beaks can provide an important, complementary, and easily accessible means to support seafood provenance and traceability, whilst integrating anthropogenic and/or geological gradients.

Composition of organic matter‑iron‑phosphorus associations in sediments of algae- and macrophyte-dominated zones in Lake Taihu

The biogeochemical cycles of phosphorus (P) and organic matter (OM) are significantly influenced by iron (Fe) through forming OM-Fe-P associations. The sources and compositions of organic matter in the two typical states (algae-dominated and macrophyte-dominated) of shallow lakes are different, which could modulate the formation of OM-Fe-P associations and influence the internal loadings of P in lakes. In this study, OM and P bound with Fe were extracted from the sediments of algae-dominated zone (A-zone) and macrophyte-dominated zone (M-zone) in Lake Taihu via the citrate-bicarbonate-dithionite (CBD) reduction method, and were analyzed to elucidate the differences in OM-Fe-P associations between the two zones. The results showed that OM-Fe-P associations in the sediments of M-zone had higher a molar C/Fe ratio but a lower molar P/Fe ratio. Four components identified by excitation-emission matrix fluorescence coupled with parallel factor analysis (EEM-PARAFAC) in OM-Fe-P associations in the two zones were all humic-like substances and the relative abundance of the corresponding PARAFAC components had no significant difference between the two zones. However, the total fluorescence intensity of the humic-like components was higher in M-zone than those in A-zone. This could be attributed to the higher aromaticity of OM produced from macrophyte than that from algae, as evidenced by the difference in the easily-desorbed OM between two zones. In M-zone, high aromatic compounds which are preferentially associated with Fe, could be more produced from macrophyte plants than from algae and resulted in the higher C/Fe molar ratio. The higher C/Fe molar ratio (2.1 ± 0.9) of OM-Fe-P associations in M-zone reflected that more OM originated from macrophyte could be involved in the coprecipitation with Fe (III) and OM-Fe-P associations in M-zone were probably more resistant to the microbial and chemical reduction. Our results from the real situation confirmed the mechanism related to the role of OM composition in the reduction of OM-Fe-P associations to explain the lower internal loading of P in M-zone than that in A-zone.

Nano and micro lupin protein-grape seed extract conjugates stabilizing oil-in-water emulsions

Conjugation of plant proteins with phenolic compounds has been an efficient strategy to improve the limited capacity of this kind of protein to stabilize emulsions. In this sense, the stabilizing capacity of lupin protein-grape seed extract conjugates in the form of nano (NC) or micro (MC) particles was evaluated in oil-in-water emulsions. Covalent binding between lupin protein isolate (LPI) and grape seed extract (GSE) was confirmed by quantification of total free sulfhydryl groups and fluorescence assays in both NC and MC for all GSE concentrations tested (0.04–1.2 wt%). The main differences between MC and NC were the larger size and surface hydrophobicity of the former, reflecting different stabilizing capacities. The droplet size and microstructure of all emulsions after 28 days of storage suggest that NC- and MC-stabilized emulsions were highly stable to coalescence. However, NC produced emulsions with a higher tendency to flocculation. Increasing the GSE concentration in the conjugates attenuated the droplet flocculation, resulting in emulsions with improved kinetic stability. Results indicate that the dominant stabilizing mechanism of the conjugates is Pickering-type, although others may occur simultaneously. Indeed, MC produced emulsions with better stability with no visible creaming and more complex rheological properties. A gel-like behavior (G’>G’’) was observed from oscillatory rheology, which was considered an additional stabilizing mechanism. In contrast, NC produced emulsions with predominantly liquid-like behavior (G’’>G’). Shear resistance was evaluated after homogenization of the emulsions, showing that the 3D network formed in the emulsions was rapidly recovered, including the highly structured MC-stabilized emulsions. Different properties of emulsions stabilized by NC or MC indicate potential varied applications in the food industry. The conjugation of GSE with LPI showed improvements in the overall stability of the emulsions, demonstrating the great potential for usage of these conjugates in the formulation of highly stable emulsion-based systems, even using a protein of limited stabilizing capacity.

Calcium signaling consequences of RyR2-S4938F mutation expressed in hiPSC-CMs.

Ryanodine receptor 2 (RyR2) is the major Ca2+ release channel of cardiac sarcoplasmic reticulum (SR) that regulates the rhythm and strength of the heartbeat, but its malfunction generates arrhythmia leading to heart failure. Here we report on a catecholaminergic polymorphic ventricular tachycardia (CPVT1)-associated RyR2 mutation (S4938F) located in RyR2 carboxyl-terminal that was expressed in human induced pluripotent stem cells derived cardiomyocytes (hiPSC-CMs) using CRISPR/Cas9 gene-editing. Ca2+ signaling and electrophysiological properties of beating cardiomyocytes carrying S4938F mutation were studied using total internal fluorescence microscopy (TIRF) and patch clamp technique. ICa densities measured either by depolarizations to zero mV or repolarizations from +100 mV to −50 mV in mutant hiPSC-CMs were comparable to WT cells, but their accompanying Ca2+-transients detected by Fura-2 (cytosolic Ca2+) or ER-GCaMP6 (SR Ca2+ release) were significantly suppressed, suggesting ICa induced Ca2+ release (CICR) was compromised. Even though 5mM caffeine triggered Fura-2 or ER-GCaMP6 signals and the frequency of spontaneously generated calcium transients of mutant cells were comparable to those of WT cells, the 4-CmC triggered cytosolic or SR Ca2+ signals and magnitude of spontaneously occurring Ca2+-transients were significantly suppressed. Spontaneously occurring Ca2+ sparks in S4938F hiPSC-CMs had longer durations but their frequency was comparable to those of WT cells. Isoprenaline increased the beating frequency that led to arrhythmic beating and cessation of excitability. Our data suggests that S4938F-RyR2 mutation does not significantly affect Ca2+content of SR, but decreases its Ca2+ sensitivity resulting in suppressed ICa-gated and spontaneous Ca2+ release from SR. Supported by NIH grant R01HL153504.

Assessing testicular germ cell DNA damage in the comet assay; introduction of a proof-of-concept.

The in vivo comet assay is widely used to measure genotoxicity; however, the current OECD test guideline (TG 489) does not recommend using the assay to assess testicular germ cells, due to the presence of testicular somatic cells. An adapted approach to specifically assess testicular germ cells within the comet assay is certainly warranted, considering regulatory needs for germ cell-specific genotoxicity data in relation to the increasing global production of and exposure to potentially hazardous chemicals. Here, we provide a proof-of-concept to selectively analyze round spermatids and primary spermatocytes, distinguishing them from other cells of the testicle. Utilizing the comet assay recordings of DNA content (total fluorescence intensity) and DNA damage (% tail intensity) of individual comets, we developed a framework to distinguish testicular cell populations based on differences in DNA content/ploidy and appearance. Haploid round spermatid comets are identified through (1) visual inspection of DNA content distributions, (2) setting DNA content thresholds, and (3) modeling DNA content distributions using a normal mixture distribution function. We also describe an approach to distinguish primary spermatocytes during comet scoring, based on their high DNA content and large physical size. Our concept allows both somatic and germ cells to be analyzed in the same animal, adding a versatile, sensitive, rapid, and resource-efficient assay to the limited genotoxicity assessment toolbox for germ cells. An adaptation of TG 489 facilitates accumulation of valuable information regarding distribution of substances to germ cells and their potential for inducing germ cell gene mutations and structural chromosomal aberrations.

Properties and metal binding behaviors of sediment dissolved organic matter (SDOM) in lakes with different trophic states along the Yangtze River Basin: A comparison and summary

The nature of sediment dissolved organic matter (SDOM) can reflect the environmental background, nutritional status and human activities and is an important part of lakes. The differences in the binding capacity of heavy metals and organic matter in lake sediments with different trophic states at the catchment scale and the mechanism of the differences in binding are still unclear. To solve this problem, we collected bulk SDOMs (< 0.7 μm) from 6 respective lakes (from upstream to downstream) in the Yangtze River Basin (YRB) to qualitatively and quantitatively characterize their properties and metal binding behaviors using excitation-emission matrix spectroscopy combined with parallel factor analysis (EEM-FARAFAC) and two-dimensional correlation spectroscopy of synchronous fluorescence spectroscopy and Fourier transform infrared spectroscopy (2D-SF-COS and 2D-FTIR-COS). The results showed that sediment dissolved organic carbon (SDOC) was mainly enriched in low molecular weight (LMW: < 1 kDa) fractions. The total fluorescence intensity (Fmax) of SDOM from upstream was larger than that from downstream (p = 0.033), and humic-like fluorophores were dominant in these lakes. The Fmax of sediment humic-like components (C1+C2) was closely related to the trophic levels of the lakes. Protein-like substances and oxygen-containing functional groups (C−OH, C=O, and C−O) were preferred in the reaction between SDOM and copper (Cu2+) or cadmium (Cd2+), while a unique binding path was exhibited in the moderately eutrophic DCL. In terms of fluorophore types, higher Cu2+-binding abilities (LogKCu) were observed in the humic-like matter for the lakes in the upper reaches and tryptophan-like matter for the lakes from the midstream and downstream areas of the YRB. Although Cd2+ complexed only with humic-like matter, LogKCd was higher than LogKCu. In terms of molecular weight (MW), the LogKCu/Cd of components were enhanced after MW fractionation. The HMW (0.7 μm – 1 kDa) components possessed higher LogKCu in most lakes (except for CHL and C4). The different fluorophores and molecular weight fractions in SDOM make an important contribution to reducing the ecological risks of heavy metals in lakes.

Light trapping by porous TiO2 hollow hemispheres for high efficiency photoelectrochemical water splitting.

Photocatalytic water splitting has recently received increasing attention as a green fuel source. The controlled nano-geometry of the photocatalytic material can improve light harvesting. In this study, as a proof of concept, hollow hemisphere (HHS)-based films of TiO2 material were created by a conventional electrospray method and subsequently applied for photoelectrochemical (PEC) water splitting. To preserve the morphology of the HHS structure, a hydrolysis precipitation phase separation method (HPPS) was developed. As a result, the TiO2 HHS-based thin films presented a maximum PEC water splitting efficiency of ca. 0.31%, almost two times that of the photoanode formed by TiO2 nanoparticle-based films (P25). The unique morphology and porous structure of the TiO2 HHSs with reduced charge recombination and improved light absorption are responsible for the enhanced PEC performance. Light scattering by the HHS was demonstrated with total reflection internal fluorescence microscopy (TRIFM), revealing the unique light trapping phenomenon within the HHS cavity. This work paves the way for the rational design of nanostructures for photocatalysis in fields including energy, environment, and organosynthesis.

Evaluation of Antifungal Potential of Indigenous Plant Extracts against Grey Mould and HPLC And LC-MS Based Identification of Phytochemical Compounds in Polygonum amplexicaule D. Don Extracts

Grey mould disease is one of the highly destructive post-harvest strawberry disease caused by the fungus Botrytis cinerea. Several synthetic compounds are being used against B. cinerea but due to resistance development by synthetic fungicide many alternative management strategies have explored nowadays. In this study, antifungal potential of indigenous plant extracts against grey mould was evaluated and amongst these plant extracts HPLC and LC-MS based identification of phytochemical compounds in Polygonum amplexicaule was also done. In this regards, firstly in vitro evaluation of the antifungal properties of twelve plant extracts was undertaken against B. cinerea using fungal growth medium, of which five plants extracts (P. amplexicaule, T. vulgaris, D. viscosa S. nigrum and E. globules) indicated the percent mycelia inhibition in fungal growth is greater than 75% which were then used for in vivo experiment. P. amplexicaule showed (81%) the highest growth inhibition in vitro as well as in in vivo (80%) activity during storage conditions (on strawberry fruits) against the fungus. HPLC analysis of methanolic extracts of P. amplexicaule showed, the total phenolics 6.176 (µg GAE/mg SW) including 0.157 (µg GAE/mg SW) gallic acid and protocatechic acid. While total fluorescence were 1.85 (µg cate/mg SW) including catechin, procyanidin and epicatechin. Total hyrdoxycinnamates were found to be 7.696 (µg CAE/mg SW) comprising on chlorogenic acids, neochlorogenic acids and 4-caffeoylquinic acids. LC-MS based identification also showed the presence of acids like caffeic, and gallic acid. Other showed the presence of rutin, quercetin, catechin, kaemferol and myricetin. It was concluded that Polygonum amplexicaule extract has effective against grey mould amongst all indigenous plant extracts and detected known and unknown compounds from the plant are well known for antimicrobial activity. Therefore in future further investigation can carried out for synthesis of bio based fungicide from these compounds of Polygonum plant extract on commercial scale against post-harvest pathogens of strawberry.

Straightforward FeOOH nanografting of Al-based SrTiO3 perovskite material as core–shell nanoflower-like heteronanostructure with enhanced solar light-driven photodegradation capability

Extensive attention and considerable efforts have been made to construct efficient heterogeneous nanoparticulate systems for surface chemical reactions to be active in solar light-driven photodegradation. This work addresses current deficiencies of the nanoparticles-focused systems intended for visible light photodegradation by developing a newly-formulated innovative chemically-engineered multi-component system that functions as a recyclabe, nontoxic, active and inexpensive catalyst for photodegradation of tetracyclne antibiotic. Here, we show a straightforward FeOOH nanografting of Al-based SrTiO3 perovskite material as core–shell nanoflower-like heteronanostructure with enhanced solar light-driven photodegradation capability over harmful antibiotics. A persuasive surface formation mechanism is proposed based on systematic investigation of the assembly process. In-depth caracterization of structural, optical and morphological properties of the prepared samples was investigated using a series of complementary analytical techniques, such as XRD, FE-SEM, HR-TEM, synchrotron XPS, as well as hard and soft XAS in both total electron yield (TEY) and fluorescence yield (TFY). The oxygen-deficient nature of core and shell interface indicates its n-doping and the availability of free charges in core which can be either transferred to the shell or create localized absorption levels into the valence band. This study provides a real opportunity to rationally photocatalysts design with very promising performance in water treatment.

Performance of electrochemical treatment of refractory organic matter in printing and dyeing reverse osmosis concentrate

The printing and dyeing wastewater treatment process produces a reverse osmosis concentrate (ROC) that contains high organic matter content and requires proper treatment before discharge. Electrochemical oxidation was applied in this study to treat organic pollutants in the ROC. In addition, the concentration variation of phthalate esters (PAEs) in ROC was investigated. The results demonstrated that the electrochemical oxidation process could effectively reduce the organic content in ROC; the total organic carbon (TOC) removal efficiency reached 83.38% at pH 2.4 and a current density of 40 mA/cm2. Increasing the current density can markedly enhance the oxidation of the bulk organic matter (i.e., TOC, chemical oxygen demand (COD), UV254, and fluorescence). The effect of pH on the decomposition of organic materials was slightly less as than that of current density. The molecular weight (MW) distribution in the ROC indicated that the percentage of components larger than 5 kDa was approximately 10%, but decreased to 2.16% with persistent electrolysis. Further, 3D fluorescence spectroscopy suggests that the organic matter in ROC was mainly aromatic proteins (APⅡ) and soluble microbial products (SMP), which account for 25.78% and 51.39% of the total fluorescence, respectively. Notably, acidic conditions are more favorable for the breakdown of fluorescence, which is attributed to the higher oxidative capacity of·OH. The decomposition of PAEs was enhanced by lowering the pH, and degradation efficiency increased by 36.45% when the pH was lowered from 9.8 to 2.4. Qualitative analysis by gas chromatography-mass spectrometry (GC-MS) showed that the abundance of organic matter in the treated ROC was significantly decreased.

Apical-Out Enteroids as an Innovative Model for Necrotizing Enterocolitis

IntroductionNecrotizing enterocolitis (NEC) is a gastrointestinal disease of premature neonates. We previously validated a NEC enteroid model derived from human infant intestinal tissue. Typical enteroid configuration is basolateral-out (BO) without direct access to the luminal (apical) surface. Apical access is necessary to allow physiologic comparison of pathogen interaction with the intestinal epithelial barrier. We hypothesize that apical-out (AO) enteroids will provide a relevant NEC model to study this relationship. MethodsFollowing the institutional review board approval (#11610-11611), neonatal intestinal tissue was collected from surgical specimens. Stem cells were collected; enteroids were generated and grown to maturity in BO conformation then everted to AO. Enteroids were untreated or treated for 24 h with 100 μg/mL lipopolysaccharide and hypoxia. Protein and gene expression were analyzed for inflammatory markers, tight junction (TJ) proteins and permeability characteristic of NEC. ResultsApical TJ protein zonula occludens-1 and basolateral protein β-catenin immunofluorescence confirmed AO configuration. Treated AO enteroids had significantly increased messenger RNA (P = 0.001) and protein levels (P < 0.0001) of tumor necrosis factor-α compared to controls. Corrected total cell fluorescence of toll-like receptor 4 was significantly increased in treated AO enteroids compared to control (P = 0.002). Occludin was found to have significantly decreased messenger RNA in treated AO enteroids (P = 0.003). Expression of other TJ proteins claudins-1, -4 and zonula occludens-1 was significantly decreased in treated AO enteroids (P < 0.05). ConclusionsAO enteroids present an innovative model for NEC with increased inflammation and gut barrier restructuring. This model allows for a biologically relevant investigation of the interaction between the pathogen and the intestinal epithelial barrier in NEC.