Serum Components Research Articles

Comparative study of rat serum pharmacochemistry between Puerariae Lobatae Radix and Puerariae Thomsonii Radix based on UPLC-Q-TOF-MS

UPLC-Q-TOF-MS and serum pharmacochemistry were employed to study the migrating components in rat sera after intragastric administration of the water extracts of Puerariae Lobatae Radix(PLR) and Puerariae Thomsonii Radix(PTR). After the respective intragastric administration of PLR and PTR extracts, blood samples were collected from the orbital vein. The serum samples were treated by protein precipitation method with methanol and acetonitrile at a ratio of 1∶1 and then passed through Agilent ZORBAX RRHD SB-C_(18) column(3 mm×100 mm, 1.8 μm) and Agilent SB-C_(18) pre-column(3 mm×5 mm, 1.8 μm) with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase. The elution was performed at the flow rate of 0.25 mL·min~(-1), the column temperature of 40 ℃, and the injection volume of 2 μL. By comparison of the total ion chromatogram and secondary fragment ion information of PLR and PTR water extracts, PLR-and PTR-containing sera, and blank serum, we found 42 migrating components(including 17 prototype components and 25 metabolites) in the sera of rats treated with PLR and 35 migrating components(including 15 prototype components and 20 metabolites) in the sera of rats treated with PTR. Thirty-three common components were shared by the two treatments, including 13 prototype components and 20 metabolites. The differences of migrating components in the PLR-and PTR-treated rat sera provide a scientific basis for further study of the active components and quality markers of PLR and PTR.

Spurious Serum Hormone Immunoassay Results: Causes, Recognition, Management.

For over 50 years, immunoassays have been extensively used to quantitate hormones in blood, other fluids and tissues. Each assay has its own sensitivity, specificity and other analytical components. Despite the differences between commercial products, these assays provide important clinical information about hormone levels in patients. However, inaccurate results can occur because of technical issues, as well as patient-specific factors that can interfere with immunoassay hormone measurements. The latter include excessive normal blood or serum components, the presence of cross-reacting substances, extremely high levels of hormones leading to the high-dose hook effect, and interference from a variety of endogenous factors such as human antibodies that interact with the assay components or high levels of biotin in the serum from exogenous ingestion. This article briefly reviews the sources and recognition of endogenous interference, and describes methods to determine the correct serum hormone concentration.

BPIFA2 as a Novel Early Biomarker to Identify Fatal Radiation Injury After Radiation Exposure

BackgroundCurrent dosimeters cannot cope with the two tasks of medical rescue in the early stage of nuclear accident, the accurate determination of radiation exposure and the identification of patients with fatal radiation injury. As radiation can cause alterations in serum components, it is feasible to develop biomarkers for radiation injury from serum. This study aims to investigate whether serum BPIFA2 could be used as a potential biomarker of predicting fatal radiation injury in the early stage after nuclear accident.MethodsA rabbit anti-mouse BPIFA2 polyclonal antibody was prepared to detect the expression of BPIFA2. C57BL/6J female mice were exposed to total body radiation (TBI) at different dose and Partial body radiation (PBI) at lethal dose to detect the dynamic changes of BPIFA2 in serum at different time points after irradiation by Western blot assay.ResultsBPIFA2 in mice serum were significantly increased at 1–12 h post-irradiation at .5–10 Gy, and increased again significantly at 3 d after 10 Gy irradiation with associated with mortality closely. It also increased rapidly after PBI and was closely related to injury degree, regardless whether the salivary glands were irradiated.ConclusionsThe increase of serum BPIFA2 is a novel early biomarker not only for identifying radiation exposure, but also for fatal radiation injury playing a vital role in rational use of medical resources, and greater efficiency of medical treatment to minimize casualties.

Comparative study on the impact on mouse livers of different amounts of Chinese Baijiu, beer, and wine consumption

Due to chronic alcohol consumption, alcoholic liver disease (ALD) has become the second most common liver disease in the world. The study was to explore the effects of consuming different amounts of various alcoholic beverages on the liver of mice. Results showed wine contained far more polyphenols and organic acids than beer and Chinese Baijiu. When daily alcohol consumption was low (less than 100 mL/60 kg/day), the liver and serum indicators of mice treated with alcohol, beer, Chinese Baijiu, and wine were not significantly different from those in saline group. At 250 mL/60 kg/day, beer used significantly damaged the liver and caused adverse changes in serum components, while wine did not cause changes in serum, indicating that it may cause less damage due to the existence of a large number of polyphenols. When the daily consumption exceeded 500 mL/60 kg/day, all four alcoholic beverages significantly increased serum markers of hepatic injury and decreased serum total antioxidant capacity and liver antioxidant enzyme levels, resulting in liver fat accumulation and serious pathological damage. At this level, beer used caused the most damage and wine the least. This study explores the appropriate drinking amount of different alcoholic beverages and provides a scientific reference for alcohol consumption.

Analysis of Proteins, Biologics, and Nanoparticles in Biological Fluids Using Asymmetrical Flow Field-Flow Fractionation

With the increasing interest in biopharmaceuticals such as proteins, antibodies, and nucleic acids, there is a corresponding increase in the need for characterizing such components. Much effort is spent on characterization in the early drug development phases as well as during formulation development and quality control. One parameter that is commonly investigated is the size distribution of the macromolecular components to deduce if there is aggregation or degradation occurring, if conformational changes occur, or if there are interactions with excipients. While the properties of the protein drug in the buffer system or in the pharmaceutical formulation are important, possibly even more interesting are the properties of the drug once it enters the body. Size characterization of macromolecules in biological fluids has traditionally been an area hampered by the complexity of the matrix. The large amount of indigenous components can interfere with commonly applied analytical techniques for size characterization. However, the separation technique asymmetrical flow field-flow fractionation (AF4) has recently shown increasing applicability for the characterization of components in blood plasma and serum. This article reviews some aspects of applying AF4 to plasma, serum, milk, and cerebrospinal fluid in the field of analysis and characterization of proteins, biologics, and nanoparticles in biological fluids.

IPS Cell Differentiation into Brain Microvascular Endothelial Cells.

The blood-brain barrier is a tissue structure that modulates the selective entry of molecules into the brain compartment. This barrier offers protection to the brain microenvironment from toxins or any fluctuations in the composition of the blood plasma via a layer of endothelial cells connected by tight junctions and supported by pericytes and astrocytes. Disruption of the barrier can be either a cause or a consequence of central nervous system pathogenesis. Therefore, research based on understanding the structure, function, and the mechanisms of breaching the blood-brain barrier is of primary interest for diverse disciplines including drug discovery, brain pathology, and infectious disease. The following protocol describes a detailed differentiation method that uses defined serum components during stem cell culture to deliver cellular cues in order to drive the cells towards brain endothelial cell lineage. This method can be used to obtain reproducible and scalable cultures of brain microvascular endothelial cells with barrier characteristics and functionality. These endothelial cells can also be stored long term or shipped frozen.

Modulating the pharmacokinetic profile of Actinium-225-labeled macropa-derived radioconjugates by dual targeting of PSMA and albumin.

Rationale: Small 225Ac-labeled prostate-specific membrane antigen (PSMA)-targeted radioconjugates have been described for targeted alpha therapy of metastatic castration-resistant prostate cancer. Transient binding to serum albumin as a highly abundant, inherent transport protein represents a commonly applied strategy to modulate the tissue distribution profile of such low-molecular-weight radiotherapeutics and to enhance radioactivity uptake into tumor lesions with the ultimate objective of improved therapeutic outcome. Methods: Two ligands mcp-M-alb-PSMA and mcp-D-alb-PSMA were synthesized by combining a macropa-derived chelator with either one or two lysine-ureido-glutamate-based PSMA- and 4-(p-iodophenyl)butyrate albumin-binding entities using multistep peptide-coupling chemistry. Both compounds were labeled with [225Ac]Ac3+ under mild conditions and their reversible binding to serum albumin was analyzed by an ultrafiltration assay as well as microscale thermophoresis measurements. Saturation binding studies and clonogenic survival assays using PSMA-expressing LNCaP cells were performed to evaluate PSMA-mediated cell binding and to assess the cytotoxic potency of the novel radioconjugates [225Ac]Ac-mcp-M-alb-PSMA and [225Ac]Ac-mcp-D-alb-PSMA, respectively. Biodistributions of both 225Ac-radioconjugates were investigated using LNCaP tumor-bearing SCID mice. Histological examinations of selected organs were performed to analyze the occurrence of necrosis using H&E staining, DNA damage via γH2AX staining and proliferation via Ki67 expression in the tissue samples. Results: Enhanced binding to serum components in general and to human serum albumin in particular was revealed for [225Ac]Ac-mcp-M-alb-PSMA and [225Ac]Ac-mcp-D-alb-PSMA, respectively. Moreover, the novel derivatives are highly potent PSMA ligands as their KD values in the nanomolar range (23.38 and 11.56 nM) are comparable to the reference radioconjugates [225Ac]Ac-mcp-M-PSMA (30.83 nM) and [225Ac]Ac-mcp-D-PSMA (10.20 nM) without albumin binders. The clonogenic activity of LNCaP cells after treatment with the 225Ac-labeled ligands was affected in a dose- and time-dependent manner, whereas the bivalent radioconjugate [225Ac]Ac-mcp-D-alb-PSMA has a stronger impact on the clonogenic cell survival than its monovalent counterpart [225Ac]Ac-mcp-M-alb-PSMA. Biodistribution studies performed in LNCaP tumor xenografts showed prolonged blood circulation times for both albumin-binding radioconjugates and a substantially increased tumor uptake (46.04 ± 7.77 %ID/g for [225Ac]Ac-mcp-M-alb-PSMA at 128 h p.i. and 153.48 ± 37.76 %ID/g at 168 h p.i. for [225Ac]Ac-mcp-D-alb-PSMA) with favorable tumor-to-background ratios. Consequently, a clear histological indication of DNA damage was discovered in the tumor tissues, whereas DNA double-strand break formation in kidney and liver sections was less pronounced. Conclusion: The modification of the PSMA-based 225Ac-radioconjugates with one or two albumin-binding entities resulted in an improved radiopharmacological behavior including a greatly enhanced tumor accumulation combined with a rather low uptake in most non-targeted organs combined with a high excretion via the kidneys.

Response of broiler chicks fed by clove and cardamom alcoholic extracts

This study investigated the effectiveness of clove and cardamom alcoholic extracts as growth promoters on performance, carcass traits, some blood serum components, immunity parameters, intestinal microbial population and intestinal morphology of broiler chicks. To this purpose, a total of 240 one-day-old broiler chicks (Ross 308 strain) were distributed randomly into three groups in a form of completely randomized design. The experimental groups were subdivided into four replicates with 20 chicks each. The experiment was continued 6 weeks. Chicks were fed by basal diet as control, basal diet contain 300 (ml.ton) clove and basal diet contain 300 (ml.ton) cardamom alcoholic extract. The result showed that significant increase in FI and BW in clove and cardamom groups while FCR decreased significantly. Using these herbal extracts already reduced abdominal fat and bursa of fabricius percentage while liver and spleen weight percentage increased significantly. Triglyceride and cholesterol levels decreased in treatments and high-density lipoprotein HDL increased. Also, low density lipoprotein LDL level decreased significantly in clove and cardamom groups. As result reveled antibody titers (ND and SRBC) increased significantly in treated groups compared to the control. It was showed that clove and cardamom extracts had significant effects on intestinal microbial population compared to control group. Data showed that Escherichia coli population decreased while Lactobacillus increased significantly compared to the control. We could demonstrate that inclusion of clove and cardamom extracts may useful and have beneficial effects on performance, blood serum components, values of immunity parameters and intestinal morphology in experimental Ross 308 broiler chicks.

Methods to improve the accuracy of spectrophotometer determination of serum creatinine

In order to solve a series of problems that affect the accuracy of spectral quantitative analysis of complex solution components, based on the “M + N” theory, this paper proposes a system for quantitative analysis of serum components by spectroscopy. At the same time, feasible methods are used in each step of the system to effectively reduce the influence of noise and interference factors, and obtain the spectral signal of serum components with high signal-to-noise ratio, so that the model has good prediction performance. In this paper, serum creatinine was taken as the research object. This paper collects multi-location and multi-mode spectral data, and uses the wavelength elimination method to optimize the wavelength to avoid over-fitting, then, based on PLS modeling, the cubic fitting is used to optimize the model, so as to reduce the influence of spectral nonlinearity caused by the scattering characteristics of solution components. After using the above methods, the prediction ability and robustness of the model are improved. Compared with the results without these methods, the correlation coefficient of creatinine concentration of all samples predicted by the model established by using the reasonable methods is increased by 32.43%, and the root mean square error is reduced by 53.61%. The above experiments show that this article combines multi-mode spectrum, the wavelength variable optimization method, the cubic equation optimization model method and other methods to quantitatively analyze the complex solution components based on the spectrum, which can improve the accuracy and robustness of the results.

Different response of Acipenser gueldenstaedtii CRP/SAP and SAA to bacterial challenge and chronic thermal stress sheds light on the innate immune system of sturgeons

Sturgeons are chondrostean fish critically endangered due to anthropogenic loss and degradation of natural habitat and overfishing for meat and caviar production. Consequently, sturgeon aquaculture has extensively developed lately, being Russian sturgeon (Acipenser gueldenstaedtii) the second most important species reared for caviar production. However, Russian sturgeon aquaculture in subtropical countries, such as Uruguay, confronts difficulties because fish have to endure excessive summertime warm temperatures, which weaken their innate defences facilitating opportunistic infections. To address this problem, we look for identifying putative acute phase proteins (APPs), which might be robust serum biomarkers of both infection and chronic thermal stress, applied to monitoring Russian sturgeon health status in farms. We focused on the C-Reactive Protein/Serum Amyloid P (CRP/SAP) pentraxin since the pentraxin family includes well-known APPs, better characterised in mammals than fish. We identified A.gueldenstaedtii CRP/SAP (AgCRP/SAP), as a member of the universal CRP/SAP pentraxin sub-family, and studied AgCRP/SAP involvement in sturgeon response to bacterial challenge and chronic thermal stress, in comparison with A. gueldenstaedtii Serum Amyloid A (AgSAA), a previously described positive APP. Results showed that AgCRP/SAP is a constitutive serum component that remained constant upon Aeromonas hydrophila challenge and chronic thermal stress. Contrastingly, serum AgSAA was subjected to regulation by bacterial and thermal stress challenges, showing a 50-fold increase and 3-fold decline in serum levels, respectively. Overall, results highlight the potential value of AgSAA, but not of AgCRP/SAP, as a biomarker of bacterial infection and the need to continue searching for robust chronic thermal stress biomarkers in sturgeons.

Induced hemocompatibility of polyethersulfone (PES) hemodialysis membrane using polyvinylpyrrolidone: Investigation on human serum fibrinogen adsorption and inflammatory biomarkers released

When blood comes in contact with dialyzer membranes, the interfacial bioactivation reactions occurring at blood/membrane boundaries detect the gross mechanism of thrombogenicity. In the presence of biocompatible polymers, the adhesion and subsequent interactions of platelets, as well as other serum components (e.g., proteins), are altered. It delays thrombus formation. In this study, the hemocompatibility of polyvinylpyrrolidone (PVP) modified polyethersulphone (PES) hemodialysis membrane fabricated using UV-assisted photochemical synthesis was investigated. To obtain varying degrees of PVP modification on each membrane, each coated surface was aged within the viscous polymer suspension for a predetermined duration. The correlation between the membrane induced hemocompatibility with membrane roughness, chemical composition, and surface charge at different aging duration of coating. In our study, we performed a comprehensive investigations on membrane morphology, hydrophilicity and its chemical composition using Atomic Force Microscopy (AFM), Fourier Transform Infra-Red (FTIR) Spectroscopy, Nuclear Magnetic Resonance (NMR), Zeta potential, and Scanning Electron Microscopy (SEM). The coated samples were analyzed using Luminex assays for the inflammatory biomarkers of Serpin/Antithrombin-III, C5a, 1L-1α, 1L-1β, IL6, and C5b-9. Modified membranes showed significant resistance against fibrinogen adsorption compared to their bare membrane counterpart. This inherent antifouling activity against fibrinogen adsorption was observed more on modified membranes that aged longer within the PVP suspension during synthesis. A longer ageing period contributed to the enhanced PVP modification, hence, more stable polymeric hydration layers. In addition, both bare and coated membranes triggered both complement system activation and inflammatory responses, but to a mild degree in the latter case. Coated membranes at longer coating duration, around ∼4 min, had smoother surfaces and lower magnitudes of surface charge and consequently exhibited reduced release of all biomarkers (except IL-1α), indicating enhanced biocompatibility. Higher levels of C5a were observed in membranes coated at 1 and 2 min, promoting modest reduction toward the release of other components.

Speciation study and biological activity of copper (II) complexes with picolinic and 6-methylpicolinic acid with different components of blood serum of low molecular mass in KNO3 1.0 mol·L−1 at 25 °C

In the present work, the chemical speciation of ternary complexes of copper (II) with picolinic acid and 6-methylpicolinic acid and different ligands, such as, components of the blood plasma of low molecular mass (lactic acid, oxalic acid, citric acid, and phosphoric acid) were studied by measurements of emf(H) in KNO3 1.00 mol·L−1. Potentiometric studies showed a predilection towards the formation of ternary species in solution, except for the copper (II)-picolinate-phosphate systems. The biological activity of the binary and ternary complexes isolated in situ, against reactive oxygen species was studied showing a concentration-dependent effect due to a possible mechanism of electron transfer. Finally, for the complexes Cu(Pic)2 and for the ligands were studied the ligand-receptor interaction on a PI3k of human origin by molecular docking, showing that, by themselves, the ligands are not capable of interacting with the active site of the enzyme. The metallic center is fundamental to generate reversible electrostatic interactions, that can be key to the indirect hypoglycemic effect exhibited by the Cu(Pic)2 complex reported in the literature.

Huiyang Shengji decoction promotes wound healing in diabetic mice by activating the EGFR/PI3K/ATK pathway

BackgroundCommon chronic wounds include diabetic ulcers, venous ulcers, and pressure ulcers. The traditional Chinese medicine Huiyang Shengji decoction (HYSJD) has been shown to promote the healing of diabetic chronic wounds, however, its pharmacological mechanism is still unclear.PurposeThis study aimed to determine the mechanism of HYSJD in promoting the healing of diabetic chronic skin ulcers.MethodsUltra-performance liquid chromatography was combined with tandem mass spectrometry (UPLC-MS/MS) to analyze the main components of HYSJD and the absorbed components in mouse serum at 30 min after oral administration of HYSJD. db/db mouse models for chronic skin ulcers were constructed by full-thickness skin resection. Wound tissues at day 7 post wound formation were used to perform microarray analysis of growth factors and chemokine expression. GO and KEGG enrichment analysis was performed on differentially expressed proteins. ELISA assays were used to measure differential expressed cytokines in the serum and Western blot analysis was used to determine the expression levels of related pathway proteins in the skin wounds.ResultsUPLC-MS/MS analysis showed that the main chemical components of HYSJD were flavonoids, terpenes, alkaloids, phenylpropanoids, and carbohydrates. At 30 min after oral administration of HYSJD, five absorbed components were detected in the serum, these included formononetin, calycosin, hypaconitine, calycosin-7-glucoside, and sinapic acid. HYSJD was found to increase the wound healing rate in chronic skin ulcers in db/db mice at days 3, 7, and 14 post wound formation, and promote the proliferation of epidermal cells. Two proteins that were differentially expressed between the different groups, i.e., IGF-1 and EGFR, were further validated. Serum ELISA assays showed that serum EGFR in the HYSJD treatment group was significantly increased. KEGG pathway analysis suggested that the PI3K/AKT pathway involved in HYSJD promoting the proliferation of epidermal cells in chronic wounds in db/db mice. Experimental verification showed that HYSJD activated the PI3K/AKT signaling pathway in mouse wound skin.ConclusionHYSJD promotes the proliferation of epidermal cells in chronic diabetic wounds by increasing EGFR expression in the wounds and activating the PI3K/AKT signaling pathway. Our study provides an experimental basis for the pharmacological mechanism of HYSJD.

Analysis of Vitamin D Components in Serum of Minors by UHPLC-MS/MS.

Serum 25-hydroxyvitamin D [25(OH)D] concentration represents the body's reserves of vitamin D, which is mostly used by clinicians to evaluate the storage status of vitamin D in the body. The present study aimed to investigate the serum vitamin D components in different health status of minors to correctly evaluate the vitamin D storage in vivo. A total of 2,270 minors were included in the study, which was divided into healthy group (1,204 cases) and disease group (1,066 cases, including 270 short stature, 433 respiratory infections, 175 malnutrition and 188 tic disorder subjects). The levels of 25-hydroxyvitamin D2 [25(OH)D2] and 25-hydroxyvitamin D3 [25(OH)D3] were measured by UHPLC-MS/MS in all subjects, and the 25(OH)D3 activity equivalents [25(OH)D3-AE] and 25(OH)D were calculated. In addition, the 3-epi-25-hydroxyvitamin D3 [3-epi-25(OH)D3] concentrations of 278 subjects (including 147 healthy and 131 disease subjects) were measured by random sampling. 25(OH)D2, 25(OH)D3, 25(OH)D and 25(OH)D3-AE levels in disease group were significantly lower than those in healthy group (p<0.001). According to the level of 25(OH)D, the sufficiency of vitamin D [25(OH)D≥30 ng/mL] was 65.4% in healthy group and 50.5% in disease group. When the 25(OH)D2 activity was converted into 25(OH)D3-AE, 53.2% of the patients in the healthy group had sufficiency vitamin D, and 39.1% in the disease group. The 3-epi-25(OH)D3 level in the disease group was significantly lower than that in the healthy group (p<0.001). Not only the 25(OH)D, but also the both of 25(OH)D2 and 25(OH)D3 levels may overestimate the vitamin D status in subjects. For accurate evaluation, at least the serum levels of 25(OH)D2, 25(OH)D3 and 3-epi-25(OH)D3 should be determined simultaneously.

Lambda monoclonal free light chain abnormalities detected by a serum immunofixation electrophoresis assay are underrepresented by quantitative serum free light chain results

Abstract Protein and immunofixation (IFIX) electrophoresis are used to diagnose and monitor monoclonal gammopathies. While IFIX detects clonal production of intact immunoglobulins and free light chains (FLC), the latter can also be quantified using a serum free light chain (SFLC) assay, in which polyclonal antisera detects epitopes specific for free kappa (KFLC) or lambda light chains (LFLC). An abnormal KFLC: LFLC ratio (KLR) serves as a surrogate for clonality. While the SFLC assay is highly sensitive, normal LFLC (&amp;lt;2.63mg/dL) and KLR results (&amp;gt;0.26 &amp; &amp;lt;1.65) were found in samples with distinct lambda monoclonal free light chains visualized by IFIX (X-LMFLC). To investigate this discordance, contemporaneous SFLC or KLR values were evaluated for their ability to accurately classify monoclonal FLCs identified by IFIX. We performed a retrospective analysis of serum and urine IFIX (Sebia Hydrasys) and SFLC (Freelite®, Binding Site) results from our institution between July 2010 through December 2020, using R 4.0.2 and Tidyverse packages. From among 9,594 encounters in which a single monoclonal component was initially identified by IFIX, 157 X-LMFLC and 131 X-KMFLC samples were analyzed. Elevated LFLC with normal KFLC was identified in 105/157 X-LMFLC samples (67%), while both LFLC and KFLC were elevated in 42/157 samples (27%). Concordance between X-KMFLC and KFLC was markedly higher, where 122/131 samples (93%) displayed elevated kappa FLC (&amp;gt;1.94mg/dL) with normal LFLC, and only 7/131 X-KMFLC samples (5%) possessed both elevated KFLC and LFLC. The use of KLR to identify pathogenic monoclonal free light chains improved lambda concordance to 85%; however, 19/157 (12%) of X-LMFLC samples still exhibited normal KLR. High concordance of 98% was again observed for X-KMFLC with abnormal KLR. When samples were segregated according to normal or impaired renal function (eGFR &amp;gt; or ≤60mL/min/1.73m², respectively), this disparate identification of X-LMFLC and X-KMFLC by the SFLC assay persisted, suggesting that renal dysfunction (as measured by eGFR) does not underlie this phenomenon. Lastly, we corroborated the above findings in a larger sample population by examining patients with urine Bence Jones FLC identified by IFIX who had free or intact monoclonal components in serum (N=724), grouped by lambda or kappa light chain involvement. The cause(s) of the discrepant performance by the Freelite® SFLC assay, relative to the Sebia Hydrasys IFIX assay, for identifying lambda FLC components is currently unclear. Possible contributory factors include assay reference range cutoffs, other patient disease parameters, and differences in assay-specific polyclonal antisera. Future analyses of these factors will help to further characterize SFLC assay performance and elucidate how interpretation of composite serum FLC test results can be improved to better guide patient management.

Characterization and functional analysis of hybrid pearl gentian grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) complement C3 against Cryptocaryon irritans infection

Pearl gentian grouper is a new aquacultural hybrid resulted from breeding of female tiger grouper (Epinephelus fuscogutattus) and male giant grouper (E. lanceolatus). Our preliminary study found that pearl gentian grouper exhibits less susceptible to the primary infection of Cryptocaryon irritans, which is an important parasitic ciliate in marine aquaculture, indicated that pearl gentian grouper might own a strong innate immune system. Complement system play key roles in innate immunity, whether pearl gentian grouper's complement component contribute for the defensing against the C. irritans infection remain unclear. In the present study, we found that C. irritans can be immobilized by untreated serum but not heat-treated serum from pearl gentian grouper, suggested that the heat-labile components in serum are responsible for the immobilization of C. irritans. Moreover, we cloned and characterized the encoding sequence of pearl gentian grouper complement C3 (PGC3), a key component in complement system. We also found that the expression level of PGC3 was increased in infected grouper serum when compared with that of control grouper. Furthermore, the binding of PGC3 on the surface of C. irritans trophonts located on the grouper skin was detected. These data suggested that pearl gentian grouper's complement system indeed play roles in the immune response against the C. irritans infection.

Comment on "Emissive H-Aggregates of an Ultrafast Molecular Rotor: A Promising Platform for Sensing Heparin".

For heparin sensing, Mudliar and Singh developed fluorescence and absorption spectroscopic approaches by utilizing emissive H-aggregates of thioflavin T (ThT) formed upon heparin binding. It has been proposed that the methods work not only in pure aqueous solution but also in complex biological media such as human serum. However, the optical features used to detect and quantify heparin are very sensitive to the ionic strength of the solution and completely vanish at 1.45 mM NaCl. Curiously, the authors were able to determine the heparin content of 1% serum samples containing the same level of electrolyte. In addition, the experimental conditions employed for heparin detection in serum samples were substantially modified, reducing the optical path length from 1 to 0.1 cm and increasing the dye concentration by an unknown measure. ThT shows a concentration-dependent tendency for aqueous aggregation, which markedly modifies its absorption and fluorescence properties. The authors have failed to verify that spectral characteristics of the ThT-heparin system observed in pure aqueous medium remain unchanged at higher dye concentrations and in the presence of serum components. Taking these issues into consideration, the heparin detection scheme offered for serum samples cannot be reproduced.

Examination of Physiological and Morphological Differences between Farm-Bred and Wild Black-Spotted Pond Frogs (Pelophylax nigromaculatus).

Simple SummaryWe compared the health status and ecological characteristics of farm-bred and wild frogs using veterinary clinical examinations. Wild frogs showed a better nutritional status and superior traits of locomotory performance than farm-bred frogs. However, wild animals are not necessarily in good health. This comparison can help determine the condition of animals and suggest improvements to the breeding environment.Due to the decline in the population and the difficulty of in situ conservation, several anuran species are being reared in captivity. In this study, we identified physiological and morphological differences between farm-bred and wild frogs. Nine different serum components were used as indicators of osmotic pressure, homeostatic state, organ function, and nutritional status of farm-bred frogs and wild frogs, while radiographic techniques were used to visualize differences in bone mineral density and body composition ratio. Additionally, X-ray skeletal images were used for morphological analysis to estimate differences in locomotory performance between the two groups. Wild frogs harbor traits that aid in better locomotory performance than farm-bred frogs. They also have a relatively lower fat content ratio and higher calcium and phosphorus serum levels than farm-bred frogs, suggesting a difference in nutritional status. However, hepatic stress was higher in wild frogs than in farm-bred frogs. Veterinary clinical examinations allow for the identification of differences in nutritional and morphological conditions between farm-bred and wild frogs. Determining the health of animals can help improve their living conditions, eliminate conditions that can negatively affect them, and effectively manage them on farms, in zoos, and at ex situ conservation institutes.

Serum and Glomerular Complement Components as Biomarkers in the First South Asian Prospective Longitudinal Observational IgA Nephropathy Cohort (GRACE-IgANI)

Serum and Glomerular Complement Components as Biomarkers in the First South Asian Prospective Longitudinal Observational IgA Nephropathy Cohort (GRACE-IgANI)

Supramolecular organization and biological interaction of squalenoyl siRNA nanoparticles

Small interfering RNAs (siRNA) are attractive and powerful tools to inhibit the expression of a targeted gene. However, their extreme hydrophilicities combined with a negative charge and short plasma half-life counteract their use as therapeutics. Previously, we chemically linked siRNA to squalene (SQ) which self-assembled as nanoparticles (NPs) with pharmacological efficiency in cancers and recently in a hereditary neuropathy. In order to understand the siRNA-SQ NP assembly and fate once intravenously injected, the present study detailed characterization of siRNA-SQ NP structure and its interaction with serum components.From SAXS and SANS analysis, we propose that the siRNA-SQ bioconjugate self-assembled as 11-nm diameter supramolecular assemblies, which are connected one to another to form spherical nanoparticles of around 130-nm diameter. The siRNA-SQ NPs were stable in biological media and interacted with serum components, notably with albumin and LDL. The high specificity of siRNA to decrease or normalize gene expression and the high colloidal stability when encapsulated into squalene nanoparticles offer promising targeted therapy with wide applications for pathologies with gene expression dysregulation.