Serum Components Research Articles

Studies on Antifungal Properties of Methacrylamido Propyl Trimethyl Ammonium Chloride Polycations and Their Toxicity In Vitro.

The biological activity of polycations is usually associated with their biocidal properties. Their antibacterial features are well known, but in this work, observations on the antifungal properties of macromolecules obtained by methacrylamido propyl trimethyl ammonium chloride (MAPTAC) polymerization are presented. The results, not previously reported, make it possible to correlate antifungal properties directly with the structure of the macromolecule, in particular the molecular mass. The polymers described here have antifungal activity against some filamentous fungi. The strongest effect occurs for polymers with a mass of about 0.5 mDa which have confirmed activity against the multidrug-resistant species Scopulariopsis brevicaulis, Fusarium oxysporum, and Fusarium solani, as well as the dermatophytes Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton interdigitale, and Trichophyton tonsurans. In addition, this publication describes the effects of these macromolecular systems on serum and blood components and provides a preliminary assessment of toxicity on cell lines of skin-forming cells, i.e., fibroblasts and keratinocytes. Additionally, using a Franz diffusion chamber, a negligibly low transport of the active polymer through the skin was demonstrated, which is a desirable effect for externally applied antifungal drugs. IMPORTANCE Infectious diseases are a very big medical, social, and economic problem. Even before the COVID-19 pandemic, certain infections were among of the most common causes of death. The difficulties in the treatment of infectious diseases concern in particular fungal diseases, against which we have only a few classes of drugs represented by a few substances. The publication presents the preliminary results of the in vitro antifungal activity studies of four MAPTAC polymers on different fungal species and their cytotoxicity to human cells (fibroblasts and keratinocytes). The paper also compares these properties with analogous ones of two commonly used antifungal drugs, ciclopirox and terbinafine.

Fluorescence turn-on detection of human serum albumin based on the assembly of gold nanoclusters and bromocresol green.

As the most abundant protein in plasma, human serum albumin plays a vital role in physiological processes, such as maintaining blood osmotic pressure and carrying small-molecule ligands. Since the content of albumin in the human serum can reflect the status of liver and renal function, albumin quantitation is significant in clinical diagnosis. In this work, fluorescence turn-on detection of human serum albumin (HSA) had been performed based on the assembly of gold nanoclusters and bromocresol green. Gold nanoclusters (AuNCs) capped by reduced glutathione (GSH) were assembled with bromocresol green (BCG), and the assembly was used as a fluorescent probe for HSA. After BCG assembling, the fluorescence of gold nanoclusters was nearly quenched. In acidic solution, HSA can selectively bind to BCG on the assembly and recover the fluorescence of the solution. Based on this turn-on fluorescence, ratiometric HSA quantification was realized. Under optimal conditions, HSA detection by the probe possessed agood linear relationship in the range of 0.40-22.50mg·mL-1, and the detection limit was 0.27 ± 0.04mg·mL-1 (3σ, n = 3). Common coexisting components in serum and blood proteins did not interfere with the detection of HSA. This method has the advantages of easy manipulation and high sensitivity, and the fluorescent response is insensitive to reaction time.

Development of a Serum-Free Medium To Aid Large-Scale Production of Mycoplasma-Based Therapies.

To assist in the advancement of the large-scale production of safe Mycoplasma vaccines and other Mycoplasma-based therapies, we developed a culture medium free of animal serum and other animal components for Mycoplasma pneumoniae growth. By establishing a workflow method to systematically test different compounds and concentrations, we provide optimized formulations capable of supporting serial passaging and robust growth reaching 60 to 70% of the biomass obtained in rich medium. Global transcriptomic and proteomic analysis showed minor physiological changes upon cell culture in the animal component-free medium, supporting its suitability for the production of M. pneumoniae-based therapies. The major contributors to growth performance were found to be glucose as a carbon source, glycerol, cholesterol, and phospholipids as a source of fatty acids. Bovine serum albumin or cyclodextrin (in the animal component-free medium) were required as lipid carriers to prevent lipid toxicity. Connaught Medical Research Laboratories medium (CMRL) used to simplify medium preparation as a source of amino acids, nucleotide precursors, vitamins, and other cofactors could be substituted by cysteine. In fact, the presence of protein hydrolysates such as yeastolate or peptones was found to be essential and preferred over free amino acids, except for the cysteine. Supplementation of nucleotide precursors and vitamins is not strictly necessary in the presence of yeastolate, suggesting that this animal origin-free hydrolysate serves as an efficient source for these compounds. Finally, we adapted the serum-free medium formulation to support growth of Mycoplasma hyopneumoniae, a swine pathogen for which inactivated whole-cell vaccines are available. IMPORTANCE Mycoplasma infections have a significant negative impact on both livestock production and human health. Vaccination is often the first option to control disease and alleviate the economic impact that some Mycoplasma infections cause on milk production, weight gain, and animal health. The fastidious nutrient requirements of these bacteria, however, challenges the industrial production of attenuated or inactivated whole-cell vaccines, which depends on the use of animal serum and other animal raw materials. Apart from their clinical relevance, some Mycoplasma species have become cellular models for systems and synthetic biology, owing to the small size of their genomes and the absence of a cell wall, which offers unique opportunities for the secretion and delivery of biotherapeutics. This study proposes medium formulations free of serum and animal components with the potential of supporting large-scale production upon industrial optimization, thus contributing to the development of safe vaccines and other Mycoplasma-based therapies.

Effect of Different Levels of Chlorogenic Acid on Growth Performance, Immunological Responses, Antioxidant Defense, and Disease Resistance of Rainbow Trout (Oncorhynchus mykiss) Juveniles.

The current study is designed to assay the efficacy of chlorogenic acid (ChA) in the diet on growth performance, digestive enzyme activity, serum immunological, biochemical, and antioxidant variables, and mucosal immune response as well as disease resistance of rainbow trout (Oncorhynchus mykiss) juveniles. Rainbow trout juveniles received diets supplemented with different inclusion levels of ChA (0 (ctrl), 200 (CA1), 400 (CA2), 600 (CA3), and 800 (CA4) mg kg-1 diet) for 60 days. According to the findings, fish from CA3 and CA4 groups demonstrated the best results considering the final weight (FW) and weight gain (WG) (P < 0.05). Also, the group that received 600 mg kg-1 ChA-supplemented diet showed the lowest feed conversion ratio (FCR) and the highest specific growth rate (SGR) compared to other groups (P < 0.05). Moreover, the minimum survival rate (SR) was only detected in the CA4 treatment (P < 0.05). Regression analysis exhibited that rainbow trout growth indices were polynomially linked to dietary chlorogenic acid concentrations. In this regard, the optimal levels of chlorogenic acid according to growth parameters (FCR and SGR) were 0.71 and 0.62 gr kg-1 diet, respectively. The results exhibited superior performance of protease and amylase activities in CA2, CA3, and CA4 groups with the maximum amount in the group receiving 600 mg kg-1 ChA-enriched diet (P < 0.05). Serum lysozyme (LYZ), immunoglobulin (Ig), and components 3 and 4 (C3 and C4) values of CA2, CA3, and CA4 groups were significantly higher than others with the highest amount in the CA3 group (P <0.05). Additionally, serum nitroblue tetrazolium (NBT) value in the CA3 and CA4 groups and myeloperoxidase (MPO) in the CA3 group were notably more than others (P < 0.05). Moreover, the lowest aspartate aminotransferase (AST), alkaline phosphatase (ALP), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) and the highest total protein (TP) and globulin (GLO) values were observed in CA3 treatment (P < 0.05). CA2 and CA3 groups demonstrated increased serum catalase (CAT) and decreased malondialdehyde (MDA) values compared to the control while the highest CAT and lowest MDA values were observed in CA3 treatment (P < 0.05). Considering mucus immunity, the significantly maximum LYZ and protease values were demonstrated in CA2 and CA3 groups, and the highest ALP, Ig, and esterase values were demonstrated in the CA3 group. In comparison with the control, the mortality rates of the groups that received the ChA diets were remarkably (P < 0.05) lower postchallenge with Y. ruckeri, and the highest survival and relative percentage of survival (RPS) (P < 0.05) belonged to the CA3 group. Results obtained from the current study suggested ChA as a functional dietary additive to raise growth parameters, immune indices, antioxidant capacity, and resistance to disease in rainbow trout.

Human Serum Supplementation Promotes Streptococcus mitis Growth and Induces Specific Transcriptomic Responses.

Streptococcus mitis is a normal member of the human oral microbiota and a leading opportunistic pathogen causing infective endocarditis (IE). Despite the complex interactions between S. mitis and the human host, understanding of S. mitis physiology and its mechanisms of adaptation to host-associated environments is inadequate, especially compared with other IE bacterial pathogens. This study reports the growth-promoting effects of human serum on S. mitis and other pathogenic streptococci, including S. oralis, S. pneumoniae, and S. agalactiae. Using transcriptomic analyses, we identified that, with the addition of human serum, S. mitis downregulates uptake systems for metal ions and sugars, fatty acid biosynthetic genes, and genes involved in stress response and other processes related with growth and replication. S. mitis upregulates uptake systems for amino acids and short peptides in response to human serum. Zinc availability and environmental signals sensed by the induced short peptide binding proteins were not sufficient to confer the growth-promoting effects. More investigation is required to establish the mechanism for growth promotion. Overall, our study contributes to the fundamental understanding of S. mitis physiology under host-associated conditions. IMPORTANCE S. mitis is exposed to human serum components during commensalism in the human mouth and bloodstream pathogenesis. However, the physiological effects of serum components on this bacterium remain unclear. Using transcriptomic analyses, S. mitis biological processes that respond to the presence of human serum were revealed, improving the fundamental understanding of S. mitis physiology in human host conditions.

Cancer cell-derived CD69 induced under lipid and oxygen starvation promotes ovarian cancer progression through fibronectin.

Cancer tissues generally have molecular oxygen and serum component deficiencies because of poor vascularization. Recently, we revealed that ICAM1 is strongly activated through lipophagy in ovarian clear cell carcinoma (CCC) cells in response to starvation of long-chain fatty acids and oxygen and confers resistance to apoptosis caused by these harsh conditions. CD69 is a glycoprotein that is synthesized in immune cells and is associated with their activation through cellular signaling pathways. However, the expression and function of CD69 in nonhematological cells is unclear. Here, we report that CD69 is induced in CCC cells as in ICAM1. Mass spectrometry analysis of phosphorylated peptides followed by pathway analysis revealed that CD69 augments CCC cell binding to fibronectin (FN) in association with the phosphorylation of multiple cellular signaling molecules including the focal adhesion pathway. Furthermore, CD69 synthesized in CCC cells could facilitate cell survival because the CD69-FN axis can induce epithelial-mesenchymal transition. Experiments with surgically removed tumor samples revealed that CD69 is predominantly expressed in CCC tumor cells compared with other histological subtypes of epithelial ovarian cancer. Overall, our data suggest that cancer cell-derived CD69 can contribute to CCC progression through FN.

Cellular uptake of biotransformed graphene oxide into lung cells

Due to its high surface area and convenient functionalization, graphene oxide has many potential applications in biomedicine, especially as a drug carrier. However, knowledge about its internalization inside mammalian cells is still limited. Graphene oxide cellular uptake is a complex phenomenon affected by factors such as the size of the particle and modifications of its surface. Moreover, nanomaterials introduced into living organisms interact with biological fluids’ components. It may further alter its biological properties. All these factors must be considered when the cellular uptake of potential drug carriers is considered. In this study, the effect of graphene oxide particle sizes on internalization efficiency into normal (LL-24) and cancerous (A549) human lung cells was investigated. Moreover, one set of samples was incubated with human serum to determine how the interaction of graphene oxide with serum components affects its structure, surface, and interaction with cells. Our findings indicate that samples incubated with serum enhance cell proliferation but enter the cells with lesser efficiency than their counterparts not incubated with human serum. What is more affinity towards the cells was higher for larger particles.

Hidden in plain sight: How helminths manage to thrive in host blood

Parasitic helminths have evolved a plethora of elegant stratagems to regulate and evade the host immune system, contributing to their considerable persistence and longevity in their vertebrate hosts. Various mechanisms to achieve this state have been described, ranging from interfering with or actively modulating host immune responses to hiding from immune recognition. Because they damage surrounding vessels and disturb blood flow, blood-borne and blood-feeding parasites in particular must deal with much more than immune effector cells. Management of the host complement system and coagulation cascade, as well as the development of processes of hiding and masking, represent hallmarks of life in blood. Here we review recent findings on putative evasion strategies employed by blood-borne parasitic helminths, focusing on the interaction with and utilisation of host serum components by nematodes and trematodes.

The Probiotic Lactobacillus sakei Subsp. Sakei and Hawthorn Extract Supplements Improved Growth Performance, Digestive Enzymes, Immunity, and Resistance to the Pesticide Acetamiprid in Common Carp (Cyprinus carpio).

This study evaluated the impacts of the probiotic, Lactobacillus sakei (L. sakei), and the extract of hawthorn, Crataegus elbursensis, on growth and immunity of the common carp exposed to acetamiprid. Fish (mean ± SE: 11.48 ± 0.1 g) feeding was done with formulated diets (T 1 (control): no supplementation, T 2: 1 × 106 CFU/g LS (Lactobacillus sakei), T3: 1 × 108 CFU/g LS, T 4: 0.5% hawthorn extract (HWE), and T 5: 1% HWE) for 60 days and then exposed to acetamiprid for 14 days. The growth performance improved in the fish fed LS at dietary level of 1 × 108 CFU/g, even after exposure to acetamiprid (P < 0.05). Intestinal Lactobacillus sakei (CFU/g) load increased (P < 0.05), following supplementation with the probiotic-enriched diet. The LS-treated fish had increases in the activity of digestive enzymes (P < 0.05). Both LS and HWE stimulated antioxidant enzymes and immune system components in serum and mucus (alkaline phosphatase (ALP), protease, total Ig, and lysozyme) (P < 0.05). However, the changes were different depending on the kind of the supplement. The malondialdehyde (MDA) levels decreased in HWE-treated fish after acetamiprid exposure (P < 0.05). Both LS and HWE reduced the liver metabolic enzymes (LDH, ALP, AST, ALT, and LDH) in serum both before and after exposure to the pesticide (P < 0.05). However, each enzyme exhibited a different change trend depending on the type of the supplement. HWE showed a stress-ameliorating effect, as glucose and cortisol levels declined in the HWE-treated fish (P < 0.05). This study indicated the immunomodulatory impacts of LS (1 × 108 CFU/g) and HWE (at dietary levels of 0.5-1%). The probiotic showed more performance compared to HWE. However, the HWE mitigated oxidative stress more efficiently than the probiotic.

C-reactive protein - My perspective on its first half century, 1930-1982.

C-reactive protein (CRP) was discovered in 1930 in the sera of patients during the acute phase of pneumococcal pneumonia and was so named because it bound to the C-polysaccharide of the pneumococcal cell wall. During the next half century many questions raised by this discovery were answered. Phosphorylcholine was found to be the moiety of the C-polysaccharide to which CRP bound. The molecular structure of CRP was elucidated: five identical subunits arranged in cyclic symmetry, giving rise to the term pentraxin. Initially felt to be not normally present in the blood, CRP was found to be a component of normal serum in trace amounts. Its site of origin was determined to be the hepatocyte. It became clear that the presumed humoral mediator responsible for CRP induction was of leukocytic origin. Binding of CRP to its ligand activated the complement system, one of the important effector mechanisms of innate immunity. CRP was found to stimulate phagocytosis of some bacterial species via binding to Fc receptors and was found to be protective in vivo against the pneumococcus in mice. It appeared likely that a related function of CRP was clearance of necrotic tissue. CRP was recognized as being a highly evolutionary conserved molecule. Its discovery during the acute phase of pneumococcal pneumonia led to its being dubbed an acute phase protein. What we today call "the acute phase response", refers to the large number of behavioral, physiologic, biochemical, and nutritional changes that occur during inflammatory states.

Abalone Viscera Fermented with Aspergillus oryzae 001 Prevents Pressure Elevation by Inhibiting Angiotensin Converting Enzyme.

Abalone viscera, which accounts for more than 20% of the total weight of abalone, is generally regarded as waste in the food industry, and effective methods are required to utilize it productively. In this study, the viscera were fermented with Aspergillus oryzae 001 to add functionality. Fermented abalone viscera exhibited increased angiotensin I-converting enzyme (ACE) inhibitory activity and enhanced inhibition of blood pressure elevation in spontaneously hypertensive rats (SHRs). Abalone viscera administration had no significant effect on body weight, food intake, liver and kidney weights, or serum components in SHRs. ACE inhibitors specific to fermented abalone viscera were identified through extraction, fractionation, purification, and analysis. The identified substance was L-m-tyrosine, which non-competitively inhibited ACE and, in a single oral administration, significantly reduced blood pressure in SHRs compared to that in the control. This study identified that abalone viscera fermented by A. oryzae 001 has an inhibitory effect on blood pressure elevation, suggesting its potential use as a functional food. In addition, L-m-tyrosine, a unique substance in fermented abalone viscera, was isolated for the first time as a single ACE-inhibitory amino acid.

Effect of PEG Anchor and Serum on Lipid Nanoparticles: Development of a Nanoparticles Tracking Method.

Polyethylene glycol (PEG) is used in Lipid Nanoparticles (LNPs) formulations to confer stealth properties and is traditionally anchored in membranes by a lipid moiety whose length significantly impacts the LNPs fate in vivo. C18 acyl chains are efficiently anchored in the membrane, while shorter C14 lipids are quickly desorbed and replaced by a protein corona responsible for the completely different fate of LNPs. In this context, a method to predict the biological behavior of LNPs depending on the lipid-PEG dissociation was developed using the Nanoparticle Tracking Analysis (NTA) method in serum. Two formulations of siRNA-containing LNPs were prepared including CSL3 or SM-102 lipids and were grafted with different lipids-PEG (C18, C14 lipids-PEG, and Ceramide-PEG). The impact of the lipid-PEG on the interactions between LNPs and serum components was demonstrated by monitoring the mean particle size and the concentration over time. In vitro, these formulations demonstrated low toxicity and efficient gene knockdown on tumor MDA-MB-231 cells, but serum was found to significantly impact the efficiency of C18-PEG-based LNPs, while it did not impact the efficiency of C14-PEG-based LNPs. The NTA method demonstrated the ability to discriminate between the behaviors of LNPs according to serum proteins' interactions. CSL3 lipid and Cer-PEG were confirmed to have promise for LNP formulation.

In vitro culture of bovine fibroblasts using select serum-free media supplemented with Chlorella vulgaris extract

Standard cell culture practices require the addition of animal-derived serum to culture media to achieve adequate cell growth. Typically, 5–10% by volume of fetal bovine serum (FBS) is used, which accounts for a vast majority of the media cost while also imposing environmental and ethical concerns associated with the use of animal serum. Here we tested the efficacy of culturing cells by replacing serum in the media with algae extract and select additives. Using LC–MS, we compared molecular signatures of FBS to Chlorella algae extracts and identified NAD(H)/NADP(H) as common and relatively abundant features in their characteristic profiles. Bovine fibroblasts, cultured in serum-free media supplemented with C. vulgaris extract and just two growth factors plus insulin, showed significant growth with enhanced viability compared to control cells cultured without serum, albeit still lower than that of controls cultured with 10% FBS. Moreover, C. vulgaris extract enhanced cell viability beyond that of cells cultured with the two growth factors and insulin alone. These results suggest that key components in serum which are essential for cell growth may also be present in C. vulgaris extract, demonstrating that it may be used at least as a partial alternative to serum for cell culture applications.

RETRACTED: Effects of the application of a food processing-based classification system in obese women: A randomized controlled pilot study.

Background: Obesity is one of the major public health problems worldwide and contributes to the onset of many diseases, especially the ones related to the metabolic syndrome. The new Dietary Guidelines for the Brazilian population bring a new food classification based on food processing and prioritizes the consumption of fresh or minimally processed foods. Aim: This study analyzed the effects of an educational intervention on obese women, on their weight loss, quality of life, components of the metabolic syndrome and pain. Methods: Randomized controlled pilot study, including 40 obese women, randomized into 2 groups: control group and intervention group. An educational intervention with 5 biweekly meetings of 90 min based on the Dietary Guidelines for the Brazilian population (2014) was carried out involving the intervention group. Parameters related to weight loss, quality of life (SF-36), pain (McGill), bioelectrical impedance analysis, cardiorespiratory fitness, and serum and clinical components of metabolic syndrome, as well as serum concentrations of cytokines were assessed. Results: Significantly decrease of body mass, waist and hip circumferences, basal metabolic rate, extracellular water, body capacitance, and body cell mass were observed in the intervention group after 3 months. Reduction of pain and improvement in quality of life and cardiorespiratory fitness were also observed in the intervention group. There were reductions in waist circumference and glycemia, components of metabolic syndrome. Conclusions: This study showed that the educational intervention can be associated with weight loss, increase in quality of life, reduction of pain, and better metabolic syndrome parameters in obese women.

Cysteinyl leukotriene receptor 1 is a potent regulator of the endosomal-lysosomal system in the ARPE-19 retinal pigment epithelial cell line.

The endosomal-lysosomal system is central for cell homeostasis and comprises the functions and dynamics of particular organelles including endosomes, lysosomes and autophagosomes. In previous studies, we found that the cysteinyl leukotriene receptor 1 (CysLTR1) regulates autophagy in the retinal pigment epithelial cell line ARPE-19 under basal cellular conditions. However, the underlying mechanism by which CysLTR1 regulates autophagy is unknown. Thus, in the present study, the effects of CysLTR1 inhibition on the endosomal-lysosomal system are analyzed in detail to identify the role of CysLTR1 in cell homeostasis and autophagy regulation. CysLTR1 inhibition in ARPE-19 cells by Zafirlukast, a CysLTR1 antagonist, depleted the lysosomal pool. Furthermore, CysLTR1 antagonization reduced endocytic capacity and internalization of epidermal growth factor and decreased levels of the transferrin receptor, CD71. Serum starvation abolished the effect of Zafirlukast on the autophagic flux, which identifies the endocytic regulation of serum components by CysLTR1 as an important autophagy-modulating mechanism. The role of CysLTR1 in inflammation and cell stress has been exceedingly studied, but its involvement in the endosomal-lysosomal pathway is largely unknown. This current study provides new insights into basal activity of CysLTR1 on cellular endocytosis and the subsequent impact on downstream processes like autophagy.

Quince extract resists atherosclerosis in rats by down-regulating the EGFR/PI3K/Akt/GSK-3β pathway

We identified the effective components and the underlying mechanisms of Quince (Cydonia oblonga Mill, COM) extract against atherosclerosis. The effective components of COM extract were identified with UHPLC-Q-TOF-MS/MS. Network pharmacology was performed. A rat model of atherosclerosis induced by high-fat emulsion combined with vitamin D3 was established. The anti-atherosclerosis effect of COM extract was evaluated from various aspects such as blood lipid regulation, anti-oxidative stress, anti-inflammatory response, and vascular protection function. We identified 14 serum components of COM extract using UHPLC-Q-TOF-MS/MS. Through prediction, 573 targets were obtained, among which 224 targets were atherosclerosis specific targets. The key targets included GSK3β, ESR1, EGFR, and HSP90AA1. The key signaling pathway was PI3K-Akt signaling pathway. Pharmacodynamics analysis showed that COM extract reduced the levels of TC, TG, and LDL-C as well as ALT and AST, while increased the level of HDL-C. Mechanistically, COM extract significantly increased serum SOD and GSH-Px activities, but decreased MDA content in atherosclerosis rats, showing antioxidant effects. Meanwhile, COM extract significantly down-regulated the levels of pro-inflammatory factors IL-1β, IL-6, TNF-α and CRP, but up-regulated anti-inflammatory factor IL-10. Additionally, COM extract increased the levels of NO, eNOS, and 6-keto-PGF1α; whereas, decreased the levels of ET-1 and TXB2. Furthermore, COM extract significantly inhibited the mRNA and protein levels of EGFR, p-PI3K, p-AKT, GSK-3β, Bax, and Caspase-3 as well as the Bax/Bcl-2 ratio. Conclusively, COM extract exerts hypolipidemic, anti-oxidative, anti-inflammatory, anti-thrombotic and vascular endothelium protective effects on atherosclerosis rat model, which may be related to the inhibition of EGFR/PI3K/AKT/GSK-3β signaling pathway.

Human Serum Mediated Bacteriophage Life Cycle Switch in Aggregatibacter actinomycetemcomitans Is Linked to Pyruvate Dehydrogenase Complex.

Antimicrobial resistance is rising as a major global public health threat and antibiotic resistance genes are widely spread among species, including human oral pathogens, e.g., Aggregatibacter actinomycetemcomitans. This Gram-negative, capnophilic, facultative anaerobe is well recognized as a causative agent leading to periodontal diseases, as well as seriously systemic infections including endocarditis. A. actinomycetemcomitans has also evolved mechanisms against complement-mediated phagocytosis and resiliently survives in serum-rich in vivo environments, i.e., inflamed periodontal pockets and blood circulations. This bacterium, however, demonstrated increasing sensitivity to human serum, when being infected by a pseudolysogenic bacteriophage S1249, which switched to the lytic state as a response to human serum. Concomitantly, the pyruvate dehydrogenase complex (PDHc), which is composed of multiple copies of three enzymes (E1, E2, and E3) and oxidatively decarboxylates pyruvate to acetyl-CoA available for tricarboxylic acid (TCA) cycle, was found up-regulated 10-fold in the bacterial lysogen after human serum exposure. The data clearly indicated that certain human serum components induced phage virion replication and egress, resulting in bacterial lysis. Phage manipulation of bacterial ATP production through regulation of PDHc, a gatekeeper linking glycolysis to TCA cycle through aerobic respiration, suggests that a more efficient energy production and delivery system is required for phage progeny replication and release in this in vivo environment. Insights into bacteriophage regulation of bacterial fitness in a mimic in vivo condition will provide alternative strategies to control bacterial infection, in addition to antibiotics.

Material basis and mechanism of Curcuma longa tuberous roots with and without vinegar processing in treating primary dysmenorrhea

Liquid chromatography-mass spectrometry was employed to analyze the chemical components in Curcuma longa tuberous roots(HSYJ), C. longa tuberous roots processed with vinegar(CHSYJ), and rat serum after the administration. The active components of HSYJ and CHSYJ absorbed in serum were identified based on the secondary spectrum of database and literature. The targets of primary dysmenorrhea was screened out from database. The protein-protein interaction network analysis, gene ontology(GO) functional annotation, and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were performed for the common targets shared by the drug active components in serum and primary dysmenorrhea, and the component-target-pathway network was constructed. AutoDock was used to conduct molecular docking between the core components and targets. A total of 44 chemical components were identified from HSYJ and CHSYJ, including 18 absorbed in serum. On the basis of network pharmacology, we identified 8 core components(including procurcumenol, isobutyl p-hydroxybenzoate, ferulic acid, and zedoarondiol) and 10 core targets \[including interleukin-6(IL-6), estrogen receptor 1(ESR1), and prostaglandin-endoperoxide synthase 2(PTGS2)\]. The core targets were mainly distributed in the heart, liver, uterus, and smooth muscle. The molecular docking results showed that the core components were well bound to the core targets, indicating that HSYJ and CHSYJ may exert therapeutic effect on primary dysmenorrhea via estrogen, ovarian steroidogenesis, tumor necrosis factor(TNF), hypoxia-inducible factor-1(HIF-1), IL-17 and other signaling pathways. This study clarifies the HSYJ and CHSYJ components absorbed in serum, as well as the corresponding mechanism, providing a reference for further elucidating the therapeutic material basis and clinical application of HSYJ and CHSYJ.

Effect of increasing valine level in the diet on performance, egg quality, and serum components in laying quails.

This research was conducted to determine the effect of increasing dietary valine level on performance parameters, egg quality criteria, and blood and serum components. In the study, 120 female Japanese quails at the age of 10weeks (241.4 ± 8.6g) did randomly allocate to six experimental groups contained five subgroups. Treatment diets were prepared to contain 0.93% (control), 1.00%, 1.10%, 1.20%, 1.30%, or 1.40% valine. The egg mass decreased linearly with the increasing level of valine in the diet, while egg weight decreased in both linear and quadratic manner (P < 0.05). Increased levels of dietary valine increased eggshell breaking strength in cubic manner (P < 0.05). The administration of the valine to the diet caused a linear decrease in the albumin index and Haugh unit (P < 0.05). Other egg quality parameters were not affected (P > 0.05). Serum glucose and triglyceride concentrations of laying quails were reduced by the increasing dietary valine levels; the effect on serum glucose was linear, while both linear and quadratic effects were significant on triglyceride concentration (P < 0.05). White blood cell, lymphocyte, and neutrophil counts increased with the increasing dietary levels of valine (P < 0.05). Both linear and quadratic effects were significant on white blood cells and lymphocytes, while the effect on neutrophil counts was linear. However, erythrocyte and haematocrit (P < 0.05) decreased quadratically with increment in the dietary levels of valine. In conclusion, the increased valine levels in the diet adversely affected the egg weight, egg mass, and Haugh unit. but it reduced the serum triglyceride and glucose. Results may suggest that dietary valine at 0.93% level is sufficient for egg production and egg quality in laying quails, but positive effects of increased dietary valine levels on white blood cells may worth to investigate further.

Determination of endogenous sphingolipid content in stroke rats and HT22 cells subjected to oxygen-glucose deprivation by LC‒MS/MS

BackgroundStroke is the leading cause of death in humans worldwide, and its incidence increases every year. It is well documented that lipids are closely related to stroke. Analyzing the changes in lipid content in the stroke model after absolute quantification and investigating whether changes in lipid content can predict stroke severity provides a basis for the combination of clinical stroke and quantitative lipid indicators.MethodsThis paper establishes a rapid, sensitive, and reliable LC‒MS/MS analytical method for the detection of endogenous sphingolipids in rat serum and brain tissue and HT22 cells and quantifies the changes in sphingolipid content in the serum and brain tissue of rats from the normal and pMCAO groups and in cells from the normal and OGD/R groups. Using sphingosine (d17:1) as the internal standard, a chloroform: methanol (9:1) mixed system was used for protein precipitation and lipid extraction, followed by analysis by reversed-phase liquid chromatography coupled to triple quadrupole mass spectrometry.ResultsBased on absolute quantitative analysis of lipids in multiple biological samples, our results show that compared with those in the normal group, the contents of sphinganine (d16:0), sphinganine (d18:0), and phytosphingosine were significantly increased in the model group, except sphingosine-1-phosphate, which was decreased in various biological samples. The levels of each sphingolipid component in serum fluctuate with time.ConclusionThis isotope-free and derivatization-free LC‒MS/MS method can achieve absolute quantification of sphingolipids in biological samples, which may also help identify lipid biomarkers of cerebral ischemia.Graphical