Serum Components Research Articles

A cationic aggregation-induced emission luminogen for colorimetric and fluorimetric detection of heparin with a dual-read approach, stability and applicability in a 10% serum matrix

Accurate detection of heparin (Hep) is essential to ensure the safety of Hep medication in clinical treatment. Although a large number of Hep optical probes have been reported in the past decade, it is still an arduous task to design a Hep optical probe with satisfactory overall performance, such as dual-mode response, stability and applicability in high-concentration serum substrates. Herein, a double-charged cationic aggregation-induced emission (AIE) luminogen, PYNN, was rationally designed and readily synthesized for efficient colorimetric and fluorimetric detection of Hep. PYNN exhibits typical AIE characteristics and good water solubility, thus emitting faint fluorescence in aqueous media. In the presence of negatively charged Hep, PYNN molecules are immobilized on the Hep chain through electrostatic and van der Waals interactions to produce a PYNN-Hep complex, causing remarkable changes in the absorption and fluorescence spectra of PYNN. The spectral response of PYNN to Hep is highly sensitive, selective, rapid, reversible and stable to ambient temperature and storage time. The absorbance ratio of PYNN at 430 nm and 370 nm and its fluorescence intensity at 580 nm meet good linear relationships with Hep concentration, with a detection limit of 6.8 ng/mL, colorimetrically and fluorimetrically, 5.9 ng/mL. PYNN can serve as a highly efficient optical probe for the dual-readout approach of Hep levels in a 10% serum matrix without interference from serum components. Our work provides an efficient, convenient and practical tool for the rapid identification and precise detection of Hep, which is of great significance for its clinical detection.

Oxidized Low-Density Lipoprotein Links Hypercholesterolemia and Bladder Cancer Aggressiveness by Promoting Cancer Stemness.

Hypercholesterolemia is a prevalent metabolic disorder that has been implicated in the development of steroid-targeted cancers. However, the link between hypercholesterolemia and urinary bladder cancer (UBC), a non-steroid-targeted cancer, remains unresolved. Here we show that diet-induced and Ldlr deficiency-induced hypercholesterolemia enhances both UBC stemness and progression. Inhibition of intestinal cholesterol absorption by ezetimibe reversed diet-induced hypercholesterolemia and cancer stemness. As a key component in hypercholesterolemic sera, oxidized low-density lipoprotein (ox-LDL), but not native low-density lipoprotein-cholesterol or metabolite 27-hydroxycholesterol, increased cancer stemness through its receptor CD36. Depletion of CD36, ectopic expression of an ox-LDL binding-disabled mutant form of CD36(K164A), and the neutralization of ox-LDL and CD36 via neutralizing antibodies all reversed ox-LDL-induced cancer stemness. Mechanistically, ox-LDL enhanced the interaction of CD36 and JAK2, inducing phosphorylation of JAK2 and subsequently activating STAT3 signaling, which was not mediated by JAK1 or Src in UBC cells. Finally, ox-LDL levels in serum predicted poor prognosis, and the ox-LDLhigh signature predicted worse survival in patients with UBC. These findings indicate that ox-LDL links hypercholesterolemia with UBC progression by enhancing cancer stemness. Lowering serum ox-LDL or targeting the CD36/JAK2/STAT3 axis might serve as a potential therapeutic strategy for UBCs with hypercholesterolemia. Moreover, elevated ox-LDL may serve as a biomarker for UBC. SIGNIFICANCE: This study demonstrates how hypercholesterolemia-induced oxidized LDL promotes urinary bladder cancer stemness via a CD36/STAT3 signaling axis, highlighting these factors as biomarkers and potential therapeutic targets of aggressive disease.

Rapid and Sensitive SERS-Based Lateral Flow Test for SARS-CoV2-Specific IgM/IgG Antibodies.

As an immune response to COVID-19 infection, patients develop SARS-CoV-2-specific IgM/IgG antibodies. Here, we compare the performance of a conventional lateral flow assay (LFA) with a surface-enhanced Raman scattering (SERS)-based LFA test for the detection of SARS-CoV-2-specific IgM/IgG in sera of COVID-19 patients. Sensitive detection of IgM might enable early serological diagnosis of acute infections. Rapid detection in serum using a custom-built SERS reader is at least an order of magnitude more sensitive than the conventional LFAs with naked-eye detection. For absolute quantification and the determination of the limit of detection (LOD), a set of reference measurements using purified (total) IgM in buffer was performed. In this purified system, the sensitivity of SERS detection is even 7 orders of magnitude higher: the LOD for SERS was ca. 100 fg/mL compared to ca. 1 μg/mL for the naked-eye detection. This outlines the high potential of SERS-based LFAs in point-of-care testing once the interference of serum components with the gold conjugates and the nitrocellulose membrane is minimized.

Potent antitumor activity of a glutamyltransferase-derived peptide via an activation of oncosis pathway

Hepatocellular carcinoma (HCC) still presents poor prognosis with high mortality rate, despite of the improvement in the management. The challenge for precision treatment was due to the fact that little targeted therapeutics are available for HCC. Recent studies show that metabolic and circulating peptides serve as endogenous switches for correcting aberrant cellular plasticity. Here we explored the antitumor activity of low molecular components in human umbilical serum and identified a high abundance peptide VI-13 by peptidome analysis, which was recognized as the part of glutamyltransferase signal peptide. We modified VI-13 by inserting four arginines and obtained an analog peptide VI-17 to improve its solubility. Our analyses showed that the peptide VI-17 induced rapid context-dependent cell death, and exhibited a higher sensitivity on hepatoma cells, which is attenuated by polyethylene glycol but not necrotic inhibitors such as z-VAD-fmk or necrostatin-1. Morphologically, VI-17 induced cell swelling, blebbing and membrane rupture with release of cellular ATP and LDH into extracellular media, which is hallmark of oncotic process. Mechanistically, VI-17 induced cell membrane pore formation, degradation of α-tubulin via influx of calcium ion. These results indicated that the novel peptide VI-17 induced oncosis in HCC cells, which could serve as a promising lead for development of therapeutic intervention of HCC.

Highly sensitive detection of dengue biomarker using streptavidin-conjugated quantum dots

A highly sensitive immunosensor using streptavidin-conjugated quantum dots (QDs/SA) was developed to detect dengue biomarker of non-structural protein 1 (NS1) at very low concentration, so that it can probe dengue infection even in the early stage. The QDs/SA were first bound to biotinylated NS1 antibody (Ab) and the QDs/SA-Ab conjugates were then used to detect the NS1 antigen (Ag) in the Ag concentration range of 1 pM to 120 nM. The formation of QDs/SA-Ab and QDs/SA-Ab-Ag conjugates was confirmed by the measurements of field emission scanning electron microscopy (FF-SEM), field emission transmission electron microscopy (FE-TEM), dynamic light scattering (DLS), and zeta-potential. Fluorescence emission spectra of QDs/SA-Ab-Ag conjugates showed that the magnitude of fluorescence quenching was linearly proportional to the NS1 Ag concentration and it nicely followed the Stern–Volmer (SV) equation in phosphate buffer solution. However, in human plasma serum solution, the fluorescence quenching behavior was negatively deviated from the SV equation presumably due to interference by the serum component biomolecules, and it was well explained by the Lehrer equation. These results suggest that the current approach is promising because it is highly sensitive, fast, simple, and convenient, and thus it has a potential of application for point-of-care.

Drug-containing serum of rhubarb-astragalus capsule inhibits the epithelial-mesenchymal transformation of HK-2 by downregulating TGF-β1/p38MAPK/Smad2/3 pathway

Ethnopharmacological relevanceRheum palmatum L; Astragalus membranaceus (Fisch.), is referred to as ‘Dahuang, Huangqi’ in China. As an important medicinal plant, the rhizome of rhubarb and astragalus is traditionally used in the treatment of kidney diseases associated with renal failure, inflammation and tumors.Aim of the study: This study aimed to investigate the effect of a drug-containing serum of rhubarb-astragalus capsules (composed of rhubarb and astragalus) and to elucidate its mechanism in the epithelial-mesenchymal transformation of renal tubular epithelial cells. Materials and methodsEpithelial-mesenchymal transformation (EMT) of HK-2 cells was induced by TGF-β1, and rhubarb-astragalus and losartan drug-containing serum from rats, as well as SB203580 (a specific inhibitor of p38 MAPK), were used. High-performance liquid chromatography analysis was performed to determine the main components of the drug-containing serum of rhubarb-astragalus from rats. Western blotting and immunofluorescence analysis were used to determine the levels of protein expression, and real-time quantitative PCR analysis was used to detect the levels of gene expression. ResultsThe drug-containing serum of rhubarb-astragalus contained emodin (0.36 μg/ml) and danthraquinone (0.96 μg/ml). Rhubarb-astragalus significantly decreased the protein expression levels of α-SMA, FN, vimentin and N-cadherin in HK-2 cells that were increased by TGF-β1, while it significantly increased the E-cadherin protein expression level that was decreased by TGF-β1. Rhubarb-astragalus also significantly decreased the protein expression levels of TGF-β1 and p38 MAPK and the mRNA expression levels of α-SMA, vimentin, TGF-β1, p38 MAPK, Smad2 and Smad3 in HK-2 cells that were increased by TGF-β1. It is worth noting that SB203580 (a p38 MAPK inhibitor) had similar effects as rhubarb-astragalus in this study. ConclusionThe drug-containing serum of rhubarb-astragalus can inhibit EMT in HK-2 cells by downregulating the TGF-β1/p38 MAPK/Smad2/3 pathway.

Functional characterization of complement factor D on the defense against Gyrodactylus kobayashii (Monogenea) infection in goldfish (Carassius auratus)

Complement factor D (CfD), a member of serine peptidases superfamily, acts as the rate-limiting enzyme of alternative complement pathway (AP) and is important for the AP-initiated C3 activation and subsequent amplification of complement cascade. The functions of the CfD in antibacterial and antiviral have been demonstrated; however, much less is known about the anti-parasite effect of CfD in teleost fish. In this study, we firstly demonstrated that the complements from goldfish serum showed high lethal effect on Gyrodactylus kobayashii acting as the AP. The serum activity against G. kobayashii was abolished by addition of EDTA, but not by EGTA + Mg2+. Subsequently, we cloned and characterized the goldfish (Carassius auratus) CfD (CaCfD), whose open reading frame consists of 753 bp, encoding 250 amino acids for a 48 kDa polypeptide. Bioinformatics analysis results showed that CfD had only one serine protease trypsin domain, and the serine active sites were completely conserved in teleost fish during evolution. The ubiquitous expression of CaCfD in all tested tissues was displayed, with the highest expression in the liver. After G. kobayashii infection, the mRNA levels of CaCfD were significantly up-regulated in blood, liver, spleen and kidney. The hemolysis percentage of the rabbit red blood cells (RRBCs) was enhanced by serum from G. kobayashii-infected goldfish, indicating that more complement components in serum were stimulated. Besides, we confirmed that recombinant CaCfD protein ((r)CaCfD) significantly improved the RRBCs lysis mediated by the complements, probably the increase of CaCfD level in complement system. (r)CaCfD also showed high anti-parasitic effect against G. kobayashii in vitro. Taken together, this study enriches the function of CfD participating in host defense against fish parasites, and the role of the complements in the protection of fishes against gyrodactylids infection deserves further investigation.

Pomalidomide (Pom) Dexamethasone (D) With Or Without Oral Weekly Cyclophosphamide (Cy) For Lenalidomide Refractory Multiple Myeloma (LRMM): A Multicenter Randomized Phase II Trial

IntroductionPomD results in an overall response rate of 30% and median PFS of 3.8 months in patients with prior lenalidomide (L) and bortezomib (Vij et al. ASCO 2012). We thought to investigate the addition of oral weekly Cy to PomD in patients with LRMM. We have previously reported that the recommended phase II dose of Cy with standard dose Pom D was 400 mg PO D1, 8, 15. We herein report aggregated results of the phase I/II trial. PatientsEligible patients had relapsed and refractory myeloma after at least 2 prior therapies and were L refractory. Patients had a platelet count≥50,000/ mm3 and ANC≥1,000/mm3 (patients with ≥50% bone marrow plasmacytosis were allowed if platelet count ≥30,000/mm3 and ANC could be supported with GCSF during screening and therapy). In the phase II portion, patients were randomized (1:1) to receive pomalidomide 4mg PO D1-21, dexamethasone 40 mg PO D1,8,15,22 (20mg if older than 75 years) with or without oral cyclophosphamide 400 mg PO D1, 8, 15 of a 28 days cycle. Patients randomized to PomD were allowed to cross over to the PomCyD in the event of disease progression. Thromboprophylaxis was mandated with aspirin, warfarin or LMWH. Responses were assessed per the IMWG criteria with the addition of minimal response (MR, ≥25% decrease in serum component). ResultsBetween 1/2012 and 7/2013, 70 patients were enrolled (10 in the phase I portion) and 30 patients were randomized to the PomCyD and 30 to PomD. The median age was 65 years (range 44-80) and 41 (58%) were male. The median number of prior therapies was 5 (range 2-14). All patients were lenalidomide refractory and 55 (79%) were refractory to proteasome inhibitors. 19 patients (27%) had high risk cytogenetics (del 17p, t(4;14), t(14;16)) of which 12 patients (17%) had del 17p. In addition, 15 (21%) had 1q21 trisomy or tetrasomy. Aggregated responses are reported for the cohort. Of the 66 patients evaluable for response, 1 achieved a CR, 10 achieved a VGPR (15%), 21 achieved a PR (32%), 13 achieved a MR (20%) and 11 achieved SD (16%). The overall response rate (PR and better) was 48.5% and the clinical benefit rate (MR and better) was 68.2%. The median progression free survival (PFS) was 6.4 months (95% CI 2.6-10.2). Patients with ≥PR had a median duration of response of 10.4 months (95% CI 6.1-14.7). The most common grade ¾ adverse event regardless of attribution were neutropenia (30% of patients), febrile neutropenia (13%), pneumonia (13%), thrombocytopenia (11%) while grade 3/4 thromboembolic events were noted in 4%. ConclusionThe aggregate response rates (≥PR in 48%) and PFS of this all oral regimen compare favorably with published results using PomD in LRMM. Enrollment on the randomized phase II trial continues. Per protocol and to avoid alpha-spending, non-aggregated results of the two arms will be presented at the meeting. Disclosures:Baz:Sanofi: Research Funding; Karyopharm: Research Funding; Novartis: Research Funding; Bristol Myers Squibb: Research Funding; Millenium: Research Funding; Celgene: Research Funding. Off Label Use: pomalidomide dexamethasone in combination with cyclophosphamide for relapsed refractory myeloma. Alsina:Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding. Shain:Celgene: Membership on an entity's Board of Directors or advisory committees; Onyx: Membership on an entity's Board of Directors or advisory committees, Research Funding. Wolf:Celgene: Honoraria, Research Funding; Millenium: Honoraria; Onyx: Honoraria. Mahindra:millenium: Membership on an entity's Board of Directors or advisory committees. Chari:Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees; Millenium: Membership on an entity's Board of Directors or advisory committees; Onyx: Membership on an entity's Board of Directors or advisory committees. Jagannath:Millennium: Honoraria; Celgene: Honoraria.

Comprehensive investigation of pharmacodyamic material basis of Wikstroemia indica (L.) C. A. Mey. by serum pharmacochemistry and bivariate correlation analysis

In this study, the theory of serum pharmacochemistry of traditional Chinese medicine was used to analyze the constituents absorbed into serum after oral administration of Wikstroemia indica (L.) C. A. Mey. by ultra high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). The micro-liquid dilution method was used to determine the minimum inhibitory concentration of the serum containing Wikstroemia indica. The bivariate correlation analysis method was used to study the spectral-efficiency relationship between the drug-containing serum and the antibacterial activity, and find the main antibacterial active components in serum containing Wikstroemia indica. A total of 26 serum migration components were identified or speculated in the samples, including 11 prototype components and 15 metabolites. Of which, syringic acid, caffeic acid, dihydrocaffeic acid, 4-hydroxybenzoic acid, hippuric acid, 3-hydroxy-3-(4-hydroxy-3-methoxyphenyl)propanoic acid, triumbelletin, (7R)-3-hydroxy-1-methyl-2-oxo-7-(prop-1-en-2-yl)-2,3,5,6,7,8- hexahydroazulene-4-carbaldehyde and (1S,3aS,8aS)-1,3,5-trihydroxy-1,4-dimethyl-7-(propan-2- ylidene)octahydroazulen-6(1H)-one were bacteriostatic active substances. It is the first time to study the constituents in serum containing Wikstroemia indica and reveal its antibacterial pharmacodyamic material basis. The above works provide scientific reference for the in-depth study of Wikstroemia indica.

A total number of 192 male one-day-old broilers chickens were randomly divided into four treatment groups of 48 chickens. Chickens of group one fed a plain diet without any supplement (control), while the diets ib groups two, three, and four were supplemented with Whole Yeast (WY, Saccharomyces cerevisiae, 0.1%), Yeast Cell Wall (YCW, 0.3 %), and Yeast Extract (YE, 0.07

A total number of 192 male one-day-old broilers chickens were randomly divided into four treatment groups of 48 chickens. Chickens of group one fed a plain diet without any supplement (control), while the diets ib groups two, three, and four were supplemented with Whole Yeast (WY, Saccharomyces cerevisiae, 0.1%), Yeast Cell Wall (YCW, 0.3 %), and Yeast Extract (YE, 0.07 %), respectively. At the end of the experimental period (35 days), the bodyweight of chickens and the feed intake of each cage were measured, and then the feed conversation ratio was calculated. Blood samples were also collected to measure the serum components and relative spleen, bursa of Fabricius, and thymus gland. The results obtained indicated that all productive performance parameters improved in response to the feeding supplementation. Blood parameters indicated that the treated groups had a significantly higher level of serum total protein and albumin as well as significantly lower serum total lipids and cholesterol. The enzyme activities of ALT, AST, and ALP were significantly reduced by WY, YCW, and YE supplementation. The relative organ weights of the spleen, bursa of Fabricius, and thymus increased significantly in broilers fed with WY, YCW, and YE, and the highest values were observed in the chickens fed with WY. It can be demonstrated that the supplementation of WY or its derivatives in the diet of broiler chickens improves the production performance as well as the physiological and immunological parameters, and consequently produce a healthier chicken.

Antibody Surface Coverage Drives Matrix Interference in Microfluidic Capillary Immunoassays.

The performance of biosensors is often optimized in buffers, which brings inconsistencies during applications with biological samples. Current strategies for minimizing sample (matrix) interference are complex to automate and miniaturize, involving, e.g., sample dilution or recovery of serum/plasma. This study shows the first systematic analysis using hundreds of actual microfluidic immunoassay fluoropolymer strips to understand matrix interference in microflow systems. As many interfering factors are assay-specific, we have explored matrix interference for a range of enzymatic immunoassays, including a direct mIgG/anti-mIgG, a sandwich cancer biomarker PSA, and a sandwich inflammatory cytokine IL-1β. Serum matrix interference was significantly affected by capillary antibody surface coverage, suggesting for the first time that the main cause of the serum matrix effect is low-affinity serum components (e.g., autoantibodies) competing with high-affinity antigens for the immobilized antibody. Additional experiments carried out with different capillary diameters confirmed the importance of antibody surface coverage in managing matrix interference. Building on these findings, we propose a novel analytical approach where antibody surface coverage and sample incubation times are key for eliminating and/or minimizing serum matrix interference, consisting in bioassay optimization carried out in serum instead of buffer, without compromising the performance of the bioassay or adding extra cost or steps. This will help establishing a new route toward faster development of modern point-of-care tests and effective biosensor development.

Insulin-Like Growth Factor 1 (IGF-1) Signaling in Glucose Metabolism in Colorectal Cancer.

Colorectal cancer (CRC) is one of the most common aggressive carcinoma types worldwide, characterized by unfavorable curative effect and poor prognosis. Epidemiological data re-vealed that CRC risk is increased in patients with metabolic syndrome (MetS) and its serum components (e.g., hyperglycemia). High glycemic index diets, which chronically raise post-prandial blood glucose, may at least in part increase colon cancer risk via the insulin/insulin-like growth factor 1 (IGF-1) signaling pathway. However, the underlying mechanisms linking IGF-1 and MetS are still poorly understood. Hyperactivated glucose uptake and aerobic glycolysis (the Warburg effect) are considered as a one of six hallmarks of cancer, including CRC. However, the role of insulin/IGF-1 signaling during the acquisition of the Warburg metabolic phenotypes by CRC cells is still poorly understood. It most likely results from the interaction of multiple processes, directly or indirectly regulated by IGF-1, such as activation of PI3K/Akt/mTORC, and Raf/MAPK signaling pathways, activation of glucose transporters (e.g., GLUT1), activation of key glycolytic enzymes (e.g., LDHA, LDH5, HK II, and PFKFB3), aberrant expression of the oncogenes (e.g., MYC, and KRAS) and/or overexpression of signaling proteins (e.g., HIF-1, TGF-β1, PI3K, ERK, Akt, and mTOR). This review describes the role of IGF-1 in glucose metabolism in physiology and colorectal carcinogenesis, including the role of the insulin/IGF system in the Warburg effect. Furthermore, current therapeutic strategies aimed at repairing impaired glucose metabolism in CRC are indicated.

Comparison of polypeptides that bind the transferrin receptor for targeting gold nanocarriers.

The ability to target therapeutic agents to specific tissues is an important element in the development of new disease treatments. The transferrin receptor (TfR) is one potential target for drug delivery, as it expressed on many dividing cells and on brain endothelium, the key cellular component of the blood-brain barrier. The aim of this study was to compare a set of new and previously-described polypeptides for their ability to bind to brain endothelium, and investigate their potential for targeting therapeutic agents to the CNS. Six polypeptides were ranked for their rate of endocytosis by the human brain endothelial cell line hCMEC/D3 and the murine line bEnd.3. One linear polypeptide and two cyclic polypeptides showed high rates of uptake. These peptides were investigated to determine whether serum components, including transferrin itself affected uptake by the endothelium. One of the cyclic peptides was strongly inhibited by transferrin and the other cyclic peptide weakly inhibited. As proof of principle the linear peptide was attached to 2nm glucose coated gold-nanoparticles, and the rate of uptake of the nanoparticles measured in a hydrogel model of the blood-brain barrier. Attachment of the TfR-targeting polypeptide significantly increased the rates of endocytosis by brain endothelium and increased movement of nanoparticles across the cells.

Shenzhiling oral liquid protects the myelin sheath against Alzheimer's disease through the PI3K/Akt-mTOR pathway

Ethnopharmacological relevanceShenzhiling oral liquid (SZL), a traditional Chinese medicine (TCM) compound, is firstly approved by the Chinese Food and Drug Administration (CFDA) for the treatment of mild to moderate Alzheimer's disease (AD). SZL is composed of ten Chinese herbs, and the precise therapy mechanism of its action to AD is far from fully understood. Aim of the studyThe purpose of this study was to observe whether SZL is an effective therapy for amyloid-beta (Aβ)-induced myelin sheath and oligodendrocytes impairments. Notably, the primary aim was to elucidate whether and through what underlying mechanism SZL protects the myelin sheath through the PI3K/Akt-mTOR signaling pathway in Aβ42-induced OLN-93 oligodendrocytes in vitro. Materials and methodsAPP/PS1 mice were treated with SZL or donepezil continuously for three months, and Aβ42-induced oligodendrocyte OLN-93 cells mimicking AD pathogenesis of myelin sheath impairments were incubated with SZL-containing serum or with donepezil. LC-MS/MS was used to analysis the active components of SZL and SZL-containing serum. The Y maze test was administered after 3 months of treatment, and the hippocampal tissues of the APP/PS1 mice were then harvested for observation of myelin sheath and oligodendrocyte morphology. Cell viability and toxicity were assessed using CCK-8 and lactate dehydrogenase (LDH) release assays, and flow cytometry was used to measure cell apoptosis. The expression of the myelin proteins MBP, PLP, and MAG and that of Aβ42 and Aβ40 in the hippocampi of APP/PS1 mice were examined after SZL treatment. Simultaneously, the expression of p-PI3K, PI3K, p-Akt, Akt, p-mTOR, and mTOR were also examined. The expression of proteins, including CNPase, Olig2, NKX2.2, MBP, PLP, MAG, MOG, p-PI3K, PI3K, p-Akt, Akt, p-mTOR, and mTOR, was determined by immunofluorescence and Western blot, and the corresponding gene expression was evaluated by qPCR in Aβ42-induced OLN-93 oligodendrocytes. ResultsLC-MS/MS detected a total of 126 active compounds in SZL-containing serum, including terpenoids, flavones, phenols, phenylpropanoids and phenolic acids. SZL treatment significantly improved memory and cognition in APP/PS1 mice and decreased the G-ratio of myelin sheath, alleviated myelin sheath and oligodendrocyte impairments by decreasing Aβ42 and Aβ40 accumulation and increasing the expression of myelin proteins MBP, PLP, MAG, and PI3K/Akt-mTOR signaling pathway associated protein in the hippocampi of APP/PS1 mice. SZL-containing serum also significantly reversed the OLN-93 cell injury induced by Aβ42 by increasing cell viability and enhanced the expression of MBP, PLP, MAG, and MOG. Meanwhile, SZL-containing serum facilitated the maturation and differentiation of oligodendrocytes in Aβ42-induced OLN-93 cells by heightening the expression of CNPase, Olig2 and NKX2.2. SZL-containing serum treatment also fostered the expression of p-PI3K, PI3K, p-Akt, Akt, p-mTOR, and mTOR, indicating an activating PI3K/Akt-mTOR signaling pathway in OLN-93 cells. Furthermore, the effects of SZL on myelin proteins, p-Akt, and p-mTOR were clearly inhibited by LY294002 and/or rapamycin, antagonists of PI3K and m-TOR, respectively. ConclusionsOur findings indicate that SZL exhibits a neuroprotective effect on the myelin sheath by promoting the expression of myelin proteins during AD, and its mechanism of action is closely related to the activation of the PI3K/Akt-mTOR signaling pathway.

Protective effect of the ethyl acetate fraction of Qinghai-Tibet Plateau medicinal plant Chrysosplenium axillare Maxim. against ANIT-induced cholestatic liver injury in mice

BackgroundChrysosplenium axillare Maxim. has been used as a traditional Tibetan herb medicine for hepatobiliary disorder in Qinghai-Tibet Plateau for more than one thousand years. Hypothesis/purposewe hypothesized that the ethyl acetate-soluble portion extracted from C. axillare (ECA) could exert hepatoprotection effect against α-naphthylisothiocyanate (ANIT)-induced hepatotoxity and acute intrahepatic cholestasis in mice and aimed to investigate its potential mechanism. Study designmale KM mice with acute intrahepatic cholestasis induced by ANIT were orally administered with the ECA (100 and 50 mg/kg B.W.). Mice receiving vehicle (0.5% CMC-Na) served as control. Methods48 h after ANIT administration, mice were sacrificed. Blood was collected for serum enzymes and components analysis using commercial kits. Livers were removed for histopathological examination and protein preparation. The protein expression levels of farnesoid X receptor (FXR), bile-salt export pump (BSEP), multidrug resistance associated protein2 (MRP2), Na+-taurocholate co-transporting polypeptide (NTCP), small heterodimer partner (SHP), and cholesterol 7α-hydroxylase (CYP7A1) were evaluated by a western blot analysis. A phytochemical analysis was conducted to characterize the chemical constituents of ECA. Resultsoral administration of ECA at both doses (P < 0.05 or P < 0.01) can prevent ANIT-induced increases in levels of serum enzymes and components, including AST, ALT, ALP, DBIL, TBA, and TBIL and improve the liver pathologic change induced by ANIT exposure. Western blot analysis showed that the expression levels of FXR, BSEP, MRP2, NTCP, and SHP in ECA (100 and 50 mg/kg) groups were up-regulated versus model group (P < 0.05 or P < 0.01), and those of CYP7A1 in both ECA treated groups were down-regulated versus model group (P < 0.01). One new highly O-methylated flavonoids, chrysosplenoside I (1), together with fourteen analogues (2–15), was isolated from ECA and structurally identified by spectra analysis. Among them, the two abundant ingredients, chrysosplenosides A and I (11 and 1), were quantitatively determined to have a total content account for 32.2% of ECA. ConclusionECA protected against mice liver damage with acute intrahepatic cholestasis induced by ANIT via regulation of the expression of hepatic proteins involved in bile acids synthesis, transportation and re-absorption. These results confirmed the hepatoprotective efficacy of traditional Tibet herb medicine C. axillare and provided the first insight into its related mechanism as well as the potential active ingredients.

Metabolic plasticity enables lifestyle transitions of Porphyromonas gingivalis

Our understanding of how the oral anaerobe Porphyromonas gingivalis can persist below the gum line, induce ecological changes, and promote polymicrobial infections remains limited. P. gingivalis has long been described as a highly proteolytic and asaccharolytic pathogen that utilizes protein substrates as the main source for energy production and proliferation. Here, we report that P. gingivalis displays a metabolic plasticity that enables the exploitation of non-proteinaceous substrates, specifically the monocarboxylates pyruvate and lactate, as well as human serum components, for colonization and biofilm formation. We show that anabolism of carbohydrates from pyruvate is powered by catabolism of amino acids. Concomitantly, the expression of fimbrial adhesion is upregulated, leading to the enhancement of biofilm formation, stimulation of multispecies biofilm development, and increase of colonization and invasion of the primary gingival epithelial cells by P. gingivalis. These studies provide the first glimpse into the metabolic plasticity of P. gingivalis and its adaptation to the nutritional condition of the host niche. Our findings support the model that in response to specific nutritional parameters, P. gingivalis has the potential to promote host colonization and development of a pathogenic community.

Hyaluronic Acid-Based Nanocapsules as Efficient Delivery Systems of Garlic Oil Active Components with Anticancer Activity.

Diallyl disulfide (DADS) and diallyl trisulfide (DATS) are garlic oil compounds exhibiting beneficial healthy properties including anticancer action. However, these compounds are sparingly water-soluble with a limited stability that may imply damage to blood vessels or cells after administration. Thus, their encapsulation in the oil-core nanocapsules based on a derivative of hyaluronic acid was investigated here as a way of protecting against oxidation and undesired interactions with blood and digestive track components. The nuclear magnetic resonance (1H NMR) technique was used to follow the oxidation processes. It was proved that the shell of the capsule acts as a barrier limiting the sulfur oxidation, enhancing the stability of C=C bonds in DADS and DATS. Moreover, it was shown that the encapsulation inhibited the lysis of the red blood cell membrane (mainly for DADS) and interactions with serum or digestive track components. Importantly, the biological functions and anticancer activity of DADS and DATS were preserved after encapsulation. Additionally, the nanocapsule formulations affected the migration of neoplastic cells—a desirable preliminary observation concerning the inhibition of migration. The proposed route of administration of these garlic extract components would enable reaching their higher concentrations in blood, longer circulation in a bloodstream, and thus, imply a better therapeutic effect.

Establishment of a reliable scheme for obtaining highly stable SERS signal of biological serum

As a rapid and non-destructive biological serum detection method, SERS technology was widely used in the screening and medical diagnosis of various diseases by combining the analysis of serum SERS spectrum and multivariate statistical algorithm. Because of the high complexity of serum components and the variability of SERS spectra, which often resulted in the phenomenon that the SERS spectrum of the same biological serum was significantly different due to the different test conditions. In this experiment, through the dilution treatment of the serum and the systematic test of the serum of all concentration gradients with lasers of wavelength of 785, 633 and 532 nm, the most suitable conditions for detecting the serum were investigated. The experimental results showed that only when the serum is diluted to low concentration (10 ppm), the SERS spectrum with high reproducibility and stability could be obtained, furthermore, the low concentration serum had weak tolerance to laser, and 532 nm laser was not suitable for serum detection. In this paper, a set of test scheme for obtaining highly stable serum SERS spectra was established by using high-performance gold nanoparticles (Au NPs) as the active substrate of SERS. Through comparative analysis of SERS spectrum of serum of normal people and cervical cancer, the reliability of the established low-concentration serum test program was verified, as well as its great potential advantages in disease screening and diagnosis.

Reference Intervals in Combined Veterinary Clinical Examinations of Male Black-Spotted Pond Frogs (Pelophylax nigromaculatus).

Simple SummaryThe reference intervals (RIs) of immunity, serum components, bone mineral density (BMD), and body composition in 151 males of Pelophylax nigromaculatus were established. These analyses are easily replicable and can safely and accurately diagnose the physiological condition of animals. The use of combined examination allows for the establishment of a successful conservation strategy through the identification of conservation problems in many vertebrate groups.In conservation physiology, analyzing the physiological response of an organism to understand its ability to adapt to environmental changes is a key technique in establishing a successful conservation strategy. Veterinary clinical examinations determine the physiological condition of animals accurately and safely, and this examination is synergistic when combined. The accuracy and safety of a clinical examination makes it advantageous for use in amphibians with high species diversity and numerous endangered species. However, it is necessary to establish a reference interval (RI) for precise interpretations and identification of animals with abnormalities through individual unit testing. We have established RIs for the immunity, serum components, bone mineral density (BMD), and body composition of black-spotted pond frogs (Pelophylax nigromaculatus). Black-spotted pond frogs are a common species and are widely distributed in East Asia, with suitable characteristics for environmental monitoring. Serum was extracted from 151 male frogs to establish the RI for bacterial killing ability in order to represent immunity. We also used the serum to establish an RI of ten additional serum components to determine the nutritional status, organ function status, body osmotic pressure, and homeostasis conditions. The BMD and three body composition measures for diagnosing food intake status and nutritional condition were measured using dual energy X-ray absorptiometry. The RI was recorded as the mean ± standard deviation, median, first (25%) to third (75%) quantile range, 95% confidence interval of the mean and median, and the 95% percentile (2.5%–97.5% range) of all components. The use of combined clinical veterinary examinations aids our understanding of the physiological conditions of an individual according to biotic and abiotic factors on a complex spatiotemporal scale in an ecosystem.

Evaluation of dietary inclusion of Bunium persicum, Bunium persicum essential oil on growth, immune components, immune‐related gene expressions and resistance to Aeromonas hydrophila , in rainbow trout, Oncorhynchus mykiss

The immune-enhancing and antibacterial properties of Black cumin, Bunium persicum have been reported in the literatures. The present work examined the effects of black cumin essential oil (BEO) administration on immunity and resistance of rainbow trout, Oncorhynchus mykiss against the pathogen, Aeromonas hydrophila. To this end, fish were fed different levels of BEO [0%, 1% (10 g/kg diet), 2% (20 g/kg diet) and 3% (30 g/kg diet)] for 60 days. At the end of feeding trial, the growth parameters and immunological components of mucus and serum assayed and then fish challenged with A. hydrophila for an additional 10 days. No significant changes were observed in growth parameters (FCR, total weight gain, total final weight) between experimental groups after feeding period (p > 0.05). The BEO in 2% and 3% levels enhanced the serum and mucosal levels of lysozyme and bactericidal activity (p < 0.01). The serum immunoglobulin (Ig) levels, alternative complement activity (ACH50) and peroxidase activity elevated in fish administered 3% BPO (p < 0.01). Total albumin in blood, total protein in mucus and blood increased in all BPO-administered fish (p < 0.01). The mucosal protease and ALP activities elevated in fish fed 2% and 3% BPO (p < 0.01). The immune-related genes (iNOS and C3) were up-regulated in BEO-administered fish, with maximum expression in fish fed 3% BEO (p < 0.01). After the challenge with A. hydrophila, the IL-1b and TNFα expression significantly elevated in all groups (p < 0.01), although it was less pronounced in the EBO administrated fish. The diets containing 2% and 3% BEO significantly decreased the fish mortality following the challenge with A. hydrophila. In addition, less hepatocyte necrosis was observed in these treatments compared with the control group. Based on GC–Mass analysis of the essential oil, some components with antioxidant and antibacterial properties such as cuminaldehyde, ρ-cymene, limonene and γ-terpinene were recognized. In conclusion, the results of the present study demonstrated that dietary BEO can enhance immunity and increase the resistance of the rainbow trout against A. hydrophila.