Complex Blood Research Articles

A novel method for characterizing cell-cell interactions at single-cell resolution reveals unique signatures in blood T cell-monocyte complexes during infection.

Communication between immune cells through direct contact is a critical feature of immune responses. Here, we developed a novel high-throughput method to study the transcriptome and adaptive immune receptor repertoire of single cells forming complexes without needing bioinformatic deconvolution. We found that T cells and monocytes forming complexes in blood during active tuberculosis (TB) and dengue hold unique transcriptomic signatures indicative of TCR/MCH-II immune synapses. Additionally, T cells in complexes showed enrichment for effector phenotypes, imaging and transcriptomic features of active TCR signaling, and increased immune activity at diagnosis compared to after anti-TB therapy. We also found evidence for bidirectional RNA exchange between T cells and monocytes, since complexes were markedly enriched for "dual-expressing" cells (i.e., co-expressing T cell and monocyte genes). Thus, studying immune cell complexes at a single-cell resolution offers novel perspectives on immune synaptic interactions occurring in blood during infection.

Study of nanolayer on red blood cells as drug carrier in an artery with stenosis

Abstract This article discusses a novel idea from cell therapy in which nanoparticles (NPs) are adsorbed on red blood cells (RBCs). RBCs serve as a drug carrier for NPs or nanodrugs adsorbed on the cell membrane of RBC. For the purpose of examination, Fe 3 O 4 {{\rm{Fe}}}_{3}{{\rm{O}}}_{4} NPs are adsorbed on RBCs, collectively called NP-RBC complex. RBCs being a natural vascular carrier, have high transfusion rates and biocompatibility. This mathematical study provides a basis to attempt nanodrug delivery via RBCs, as carriers for nanodrugs, to the stenosed sites in an artery. The mathematical model is developed for an artery with stenosis and a catheter that regards the temperature and velocity of the NP-RBC complex. Catheter coated with the NP-RBC complex is inserted into the lumen of the stenosed artery. The mathematical problem is solved numerically using Bernstein polynomials. The physical features were discussed through graphs plotted using MATLAB. The influence of parameters such as volume fraction, radius of the NP-RBC complex in blood, and the thickness of the nanolayer on RBCs was studied. A noticeable outcome states that the nanolayer of optimum thickness about 50–40 nm is suitable for this purpose. Thus, this is an attempt to study the delivery of NPs adsorbed on the surface of RBCs to develop newfangled strategies in nanomedicine bearing high precision and efficiency.

Characterizing blood hysteresis via tensorial thixo-elasto-viscoplastic modeling

A characteristic feature of human blood rheology is a distinctive stress hysteresis during shear ramp up in the shear rate from zero, followed by a ramp back to zero. This is a result of the fact that human blood has a longer characteristic time of shear-induced rouleaux breakdown compared to the shear aggregation of the rouleaux. We demonstrate this telltale phenomenon of human blood rheology during the triangle ramp, time-dependent change in the shear rate. The unique hysteresis data are then used along with steady state data to fit parameters of a recently published thixo-elasto-viscoplastic rheological model, the tensorial enhanced structural stress thixotropic-viscoelastic (t-ESSTV) model. These best-fit parameter values from the hysteresis ramps are then used to predict step-up/down in shear, small amplitude oscillatory shear, uni-directional large amplitude oscillatory shear, and large amplitude oscillatory shear flow. Additionally, correlations between the calculated fitting parameters and physiological data are analyzed to inform the interpretation of model behavior in physical terms. The fit adherence to the triangle ramp and rheological hysteresis data are then evaluated alongside recently developed techniques to assess thixotropy via hysteresis loop area, indicating the efficacy of the t-ESSTV model in potentially predicting blood's complex characteristics in useful ways for future use in modeling blood flows under a variety of mechanical and biological conditions and predicting pathologies.

Performance of IS6110-LAMP assay for detection of Mycobacterium tuberculosis complex in blood and urine samples from patients with extrapulmonary tuberculosis

BackgroundThe diagnosis of extrapulmonary tuberculosis (EPTB) shows numerous difficulties because of non-specific symptomatology and low sensitivity of conventional methods. Loop-mediated isothermal amplification (LAMP) is a fast and low-cost technique, which can amplify under isothermal conditions an amount of target DNA copies into approximately a billion copies. ObjectiveThe present study aimed to evaluate a IS6110-LAMP system for Mycobacterium tuberculosis detection in blood and urine samples from patients with EPTB. MethodsThe collected samples (n = 122) were stratified in two groups: Group EPTB – patient samples with confirmed EPTB (n = 61); Group non-TB – patient samples without TB (n = 61). The urine samples underwent decontamination, and the components of blood samples were separated (plasma and PBMC). DNA extractions were performed in all biological samples followed by IS6110-LAMP assay technique. The detection limit was evaluated through dilution curves (1:10) using Mtb reference strain (H37Rv) genomic DNA. FindingsThe detection limit of IS6110-LAMP was 10 fg/μL (∼10–20 bacilli/μL). The IS6110-LAMP technique sensitivity and specificity were 95.65 % and 79.25 %, respectively, with a general kappa agreement index of 0.762. Main conclusionsBased on these results, IS6110-LAMP test showed considerable diagnostic parameters, being able to aid in the speed and accuracy of the final EPTB diagnosis.

The state of the humoral and cellular links of immunity of recipient rabbits following allogeneic transfusion of erythrocyte mass

The study of activation of the humoral link of immunity of recipient rabbits following allogeneic transfusion of erythrocyte mass is important for a fundamental understanding of the formation of immunity in model species of animals. We measured the contents of B-lymphocytes, immunoglobulins of M, G, A classes, circulating immune complexes in blood of the recipient rabbits and antibody-dependent cytotoxic activity of lymphocytes. Number of B-lymphocytes in the blood of the rabbits was determined according to Bianco. Immunoglobulin content in blood serum of the rabbits was measured based on the Mancini method. The spectrophotometry method was used to measure circulating immune complexes. The antibody-dependent cytotoxic activity of lymphocytes was identified by the colorimetric method. Modeling of erythrocyte mass transfusion was performed on five clinically healthy rabbits by intravenous injection of allogeneic erythrocyte mass in the dose of 5.5 mL/kg of body weight. The materials for the study were the blood samples, gathered on the 3rd, 7th, and 23rd days after transfusion. We found that the content of B-lymphocytes in blood of the recipient rabbits increased throughout the research: on the 3rd day the content of B lymphocytes, compared with the initial state (11.4%), increased by 1.94 times and was 22.2%; on the 7th day their content increased by 2.22 times and was 25.4%; and on the 23rd day of the experiment the content of lymphocytes increased by 2.46 times, compared with the initial state, equaling 28.6%. The content of IgM on the 3rd day after transfusion of erythrocyte mass increased by 54.9% and was 2.20 ± 0.70 g/L. On the 7th day it increased by 19.0%, equaling 1.69 ± 0.44 g/L, and on the 23rd day the content of immunoglobulins decreased by 54.2% compared with the initial state (1.42 ± 0.18 g/L, intact rabbits), accounting for 0.77 ± 0.25 g/L. The content of immunoglobulins of class G on the 3rd, 7th, and 23rd days of the experiment was nearly the same as in the initial state. The content of class A immunoglobulins decreased 4.16-fold on the 3rd day of the experiment, approaching the control values afterwards. After the transfusion of erythrocyte mass to the recipient rabbits, the content of circulating immune complexes was higher compared with the initial state throughout the research: on the 3rd day the CICs content increased by 2.13 times, measuring 6.40 conventional units, and on the 7th and 23rd days it approached the initial values of the control group. On the 3rd day after transfusion, the antibody-dependent cytotoxic activity of blood lymphocytes of erythrocyte mass of the rabbits increased compared with the initial state (15.9%), measuring 17.3%; on the 7th day it was 19.4%; and on the 23rd day it increased to 27.9%. The results revealed presence of characteristic immunological changes after allogeneic transfusion of erythrocyte mass to the recipient rabbits, which were a consequence of progressive increase in the activity of the humoral link of immunity.

Placental Barrier on Cadmium Transfer from Mother to Fetus in Related to Pregnancy Complications.

As two of the most severe and common medical disorders during pregnancy, gestational diabetes mellitus (GDM) and hypertensive disorder complicating pregnancy (HDCP) cause adverse effects on placental barrier function and thus may lead to a high risk of intrauterine exposure to toxic metals from mother to fetus. This study investigates the impact of the placental barrier on the transfer of cadmium (Cd) from mother to fetus and the relationship between pregnancy complications. A total of 107 pairs of samples were collected in Kunming, China; 29 were from healthy pregnant women, and 78 were from patients with pregnancy complications. Cd was measured in each mother's placenta and maternal and umbilical cord blood. The expressions of MT and Cd-MT complex in blood and placental tissue samples were determined by enzyme-linked immunosorbent assay (ELISA). The cesarean section rate in the whole pathological group (60.7%) was higher than that in the normal group (20.7%), and the ratio of the effective barrier (ratio of maternal blood to umbilical cord blood>1) in the pathological group (74%) was lower than that in the normal group (79%). In addition, the proportion of practical placental barriers in women aged 20-25 years was 83.3%, 76.3% in women aged 26-30 years, 74.3% in women aged 31-35 years, 70% in women aged 36-40 years, and 71% in women aged 40-45 years. The Cd content in the placenta of the three pathological groups was significantly higher than that in maternal and umbilical cord blood (P<0.05), and the distribution of Cd was the same as that in the normal group. However, there was no significant difference between maternal and umbilical cord blood Cd concentrations in the pathological group. The Cd concentration in the normal group's maternal blood was significantly higher than that in cord blood (P<0.05). In addition, the expression levels of both metallothionein (MT) and Cd-MT complex in placenta is much higher than in maternal and umbilical blood, and which in normal group are significantly higher than those in pathological group. Both mothers and fetuses are at increased health risk for pregnancy disorders when maternal age, BMI, or body weight increases. Increased maternal age increases the likelihood of Cd transfer from the mother to the fetus. Pregnancy complications may induce lower expression of MT, thus reducing the Cd-MT complex in the placenta, weakening the placental barrier, and increasing the risk of Cd transfer and exposure to the fetus.

PLATELET- NEUTROPHIL COMPLEXES – DEFINITION, MECHANISMS AND IMPLICATIONS (REVIEW)

Neutrophils are innate immune cells that protect the host from infection by eliminating the invadingpathogens. In recent years, neutrophils and platelets have been shown to be active not only in inflammatoryprocesses but also in some pathological conditions, including the development of neoplastic diseases andthrombosis. The interaction between circulating platelets and neutrophils is characterised by the intensive exchangeof intracellular signals. Overexpression of some of these molecules has been reported, with increased leukocyte andplatelet activation as well as increased adhesion of blood cells to the vascular endothelium. Platelet-neutrophilinteractions are increased in thrombo-inflammatory diseases and are sensitive biomarkers for platelet activation andtargets for the development of new therapies. The crosstalk between platelets and innate immune cells promotesthrombosis, inflammation, and tissue damage.A key step in the formation of platelet-neutrophil complexes (PNCs) is the process of interaction between theadhesion molecule P-selectin (CD62P) on the platelet membrane and the PSGL-1 ligand on neutrophils. Theformation of PNCs modulates a wide range of pathological conditions, including both innate and acquired immunesystems. PNCs are involved in the pathogenesis of inflammatory diseases, malignancies and thrombotic processes.The formation of PNCs is associated with the initiation of a number of processes at the cellular and molecular levels.Malignant diseases are risk factors for the occurrence of arterial, venous and microthrombosis, which are most oftenassociated with hypercoagulable processes that form platelet aggregates. By modulating the immune system throughdirect and indirect interactions with leukocytes, platelets regulate several aspects of tumor-related pathology.Thisreview aims at summarizing the role of platelet-neutrophil interactions in the development and progression of cancerand puts its focus on cancer-related alterations of platelet and leukocyte functions and their impact on cancerpathology.The analysis and evaluation of the levels of the circulating platelet- neutrophil complexes in blood in neoplasticdiseases can be used as potential predictors of tumor progression, metastasis and the occurrence of thrombosis.

Preparation of radiolabeled erlotinib analogues and analysis of the effect of linkers

Erlotinib is a first generation epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) which was granted Food and Drug administration (FDA) approval for treatment of patients with locally advanced or metastatic NSCLC. The present study aimed at development of radiolabeled erlotinib variants as tyrosine kinase inhibitors. Three DOTA-erlotinib conjugates were prepared for radiolabeling with 177Lu. The terminal alkyne group of erlotinib was modified by performing Cu-catalyzed click chemistry and three different linkers were introduced which were then conjugated to the chelator, DOTA. The DOTA-erlotinib conjugates were characterized by 1H NMR and ESI-MS. 177Lu-DOTA-erlotinib complexes were characterized using natLu-DOTA-erlotinib conjugates. The 177Lu-complexes exhibited high in vitro stability in human serum up to 48 h. They were highly hydrophilic in nature as observed from their log Po/w values (177Lu-DOTA-propyl-Er: −2.5 ± 0.1; 177Lu-DOTA-PEG3-Er: −3.0 ± 0.1; 177Lu-DOTA-PEG6-Er: −3.3 ± 0.1). The MTT assay in A431 human epidermoid carcinoma cell lines indicates that the chemical modification at the terminal alkyne group of the erlotinib molecule does not have significant effect on its TKI property. Biodistribution studies in normal Swiss mice demonstrated fast clearance and excretion of 177Lu-labeled erlotinib complexes. These studies indicate that erlotinib variants with hydrophobic pharmacokinetic modifiers/chelators may enhance the retention of 177Lu-labeled complexes in blood thereby increasing the probability to reach EGFR-expressing tumor.

Development and validation of an LC-MS/MS method for determination of B vitamins and some its derivatives in whole blood.

Obligate symbiotic bacteria associated with the insects feeding exclusively on vertebrate blood are supposed to complement B vitamins presumably lacking in their diet. Recent genomic analyses revealed considerable differences in biosynthetic capacities across different symbionts, suggesting that levels of B vitamins may vary across different vertebrate hosts. However, a rigorous determination of B vitamins content in blood of various vertebrates has not yet been approached. A reliable analytical method focused on B vitamin complex in blood can provide valuable informative background and understanding of general principles of insect symbiosis. In this work, a chromatographic separation of eight B vitamins (thiamine, riboflavin, niacin, pantothenic acid, pyridoxine, biotin, folic acid, and cyanocobalamine), four B vitamin derivatives (niacinamide, pyridoxal-5-phosphate, 4-pyridoxic acid, and tetrahydrofolic acid), and 3 stable isotope labelled internal standards was developed. Detection was carried out using dual-pressure linear ion trap mass spectrometer in FullScan MS/MS and SIM mode. Except for vitamin B9 (tetrahydrofolic acid), the instrument quantitation limits of all analytes were ranging from 0.42 to 5.0 μg/L, correlation coefficients from 0.9997 to 1.0000, and QC coefficients from 0.53 to 3.2%. Optimization of whole blood sample preparation step was focused especially on evaluation of two types of protein-precipitation agents: trichloroacetic acid and zinc sulphate in methanol. The best results were obtained for zinc sulphate in methanol, but only nine analytes were successfully validated. Accuracy of the procedure using this protein-precipitating agent was ranging from 89 to 120%, precision from 0.5 to 13%, and process efficiency from 65 to 108%. The content of B vitamins in whole blood samples from human and various vertebrates is presented as an application example of this newly developed method.

In silico design of a TLR4-mediating multiepitope chimeric vaccine against amyotrophic lateral sclerosis via advanced immunoinformatics.

Amyotrophic lateral sclerosis (ALS) is the most prevalent motor neuron disorder worldwide. In ALS, progressing disease can result from misfolding and aggregation of superoxide dismutase-1 (SOD1) or TAR DNA-binding protein 43kDa (TDP43). An efficient immunotherapy for ALS should spare intact SOD1 while eliminating its dysfunctional variant. We utilized advanced immunoinformatics to suggest a potential vaccine candidate against ALS by proposing a model of dynamic TLR4 mediation and induction of a specific Th2-biased shift against mutant SOD1, TDP43, and TRAF6, a protein that specifically interacts with dysfunctional SOD1. SOD1, TDP43, and TRAF6 were retrieved in FASTA. Immune Epitopes Database and CTLpred suggested T/B-cell epitopes from disease-specific regions of selected antigens. A TLR4-mediating adjuvant, RS01, was used. Sequences were assembled via suitable linkers. Tertiary structure of the protein was calculated. Refined protein structure and physicochemical features of the 3D structure were verified in silico. Differential immune induction was assessed via C-ImmSim. GROningen MAchine for Chemical Simulation was used to assess evolution of the docked vaccine-TLR4 complex in blood. Our protein showed high structural quality and was nonallergenic and immune inducing. Also, the vaccine-TLR4 complex stability was verified by RMSD, RMSF, gyration, and visual analyses of the molecular dynamic trajectory. Contact residues in the vaccine-TLR4 complex showed favorable binding energies. Immune stimulation analyses of the proposed candidate demonstrated a sustained memory cell response and a strong adaptive immune reaction. We proposed a potential vaccine candidate against ALS and verified its physicochemical and immune inducing features. Future studies should assess this vaccine in animal studies.

Leishmaniasis in wild, synanthropic and domestic animals on the Island of Itamaracá, Pernambuco, Northeastern Brazil

Leishmaniasis are notifiable diseases, characterized by severe clinical manifestations, being caused by obligate intracellular protozoa, belonging to the genus Leishmania, where the species L. (L.) infantum and L. (V.) braziliensis are responsible for most cases. of leishmaniasis in Brazil. Many studies indicate the need for investigations into the exact role of animal species in the epidemiology of leishmaniasis, particularly of skunks and rodents, where the presence of these animals increases the chances of humans and other animals becoming infected. Considering the scarcity of information on the epidemiology of leishmaniasis in some regions of the State of Pernambuco, the objective of this study was to evaluate the occurrence of infection by Leishmania spp. in wild, synanthropic and domestic animals. For this, the parasitological diagnosis was carried out through exfoliative cytology of the skin of cutaneous lesions, in addition to PCR of the L. donovani and L. braziliensis complex of blood and spleen, liver and skin biopsy of rodents and skunks captured in an endemic area for leishmaniasis in the state of Pernambuco. In the locations where synanthropic animals positive for Leishmania were captured, blood, skin, bone marrow and lymph nodes were collected from the canine population. As a result, amastigote forms and DNA from the L. donovani complex were detected in 8.3% (2/24) of synanthropic animals (D. albiventris and O. nigripes). In the canine population, 29.4% (15/51) of dogs reactive for Visceral Leishmaniasis, and 6.6% were parasitologically and molecularly positive for the L. donovani complex. Thus, it is concluded that skunks, rodents and dogs participate in the transmission cycle of the L. donovani complex.

The von Willebrand factor A-1 domain binding aptamer BT200 elevates plasma levels of von Willebrand factor and factor VIII: a first-in-human trial.

von Willebrand factor (VWF) and factor VIII (FVIII) circulate in a noncovalent complex in blood and promote primary hemostasis and clotting, respectively. A new VWF A1-domain binding aptamer, BT200, demonstrated good subcutaneous bioavailability and a long half-life in non-human primates. This first-in-human, randomized, placebo-controlled, double-blind trial tested the hypothesis that BT200 is well tolerated and has favorable pharmacokinetic and pharmacodynamic effects in 112 volunteers. Participants received one of the following: a single ascending dose of BT200 (0.18-48 mg) subcutaneously, an intravenous dose, BT200 with concomitant desmopressin or multiple doses. Pharmacokinetics were characterized, and the pharmacodynamic effects were measured by VWF levels, FVIII clotting activity, ristocetin-induced aggregation, platelet function under high shear rates, and thrombin generation. The mean half-lives ranged from 7-12 days and subcutaneous bioavailability increased dose-dependently exceeding 55% for doses of 6-48 mg. By blocking free A1 domains, BT200 dose-dependently decreased ristocetin-induced aggregation, and prolonged collagen-adenosine diphosphate and shear-induced platelet plug formation times. However, BT200 also increased VWF antigen and FVIII levels 4-fold (P<0.001), without increasing VWF propeptide levels, indicating decreased VWF/FVIII clearance. This, in turn, increased thrombin generation and accelerated clotting. Desmopressin-induced VWF/FVIII release had additive effects on a background of BT200. Tolerability and safety were generally good, but exaggerated pharmacology was seen at saturating doses. This trial identified a novel mechanism of action for BT200: BT200 dose-dependently increases VWF/FVIII by prolonging half-life at doses well below those which inhibit VWF-mediated platelet function. This novel property can be exploited therapeutically to enhance hemostasis in congenital bleeding disorders.

Secrets, puzzles and mysteries of macroamylasemia

Physiological features of amylase synthesis and excretion are considered in the article, presence of other sources of amylase synthesis different from pancreas and salivary glands is emphasized. Definitions of hyperenzymemia and macroamylasemia (MAE) are given. MAE is a state characterized by presence of circulating complexes of normal serum amylase with protein or carbohydrates in blood. There are 3 types of MAE: first — classical (constant hyperamylasemia, decreased amylase level in urine, high blood concentration of macroamylase complexes); second — hyperamylasemia with slightly decreased amylase activity in urine, macroamylase/normal amylase ratio is less than in the first type; third — normal blood and urine amylase activity, low macroamylase/normal amylase ratio. Pathogenesis is explained by connection of blood amylase and acute phase protein in different inflammatory, infectious diseases, malabsorption. MAE clinical manifestations could be absent, sometimes abdominal pain is possible. Hyperamylasemia and reduced urine amylase activity are typical. MAE diagnostics means determination of macroamylase complexes in blood (chromatography, calculation of the clearance ratio of amylase and creatinine). The article presents clinical cases describing extra-pancreatic MAE in women with malignant ovarian lesions. The question of expediency of thorough diagnostic examination in asymptomatic MAE is raised, which may turn out to be a symptom of cancer. The lack of specific treatment for MAE is emphasized.

Rapid detection of Mycobacterium tuberculosis in children using blood and urine specimens.

INTRODUCTION: Laboratory and clinical features of childhood tuberculosis (TB) are non-specific and establishing an accurate diagnosis remains a challenge. This study evaluated a Single tube nested-PCR (STNPCR) to detect genomic DNA of Mycobacterium tuberculosis complex in blood and urine. METHODS: Biological samples were obtained from children (<15 years old) with clinical suspicion of pulmonary and extrapulmonary TB at public hospitals in Recife-Pernambuco, Brazil. Cultures yielded negative results in a majority of childhood TB cases, which are generally paucibacillary. A set of clinical, epidemiological, radiological, and laboratory criteria with evident clinical improvement after anti-TB treatment were frequently used to define childhood TB cases. RESULTS: Ninety children with clinical suspicion were enrolled in this study (44 with TB and 46 without TB). The pulmonary TB group had 20 confirmed cases and 46 negative controls, while the extrapulmonary TB group had 24 confirmed cases. The STNPCR showed sensitivities to pulmonary and extrapulmonary TB of 47.4% and 52.2% (blood) and 38.8% and 20% (urine), respectively. Considering the low performance of STNPCR on separate samples, we decided to perform a combined analysis (parallel sensitivity analysis) of the results from blood and urine samples. The parallel sensitivity increased to 65% in blood and 62.5% in urine. The specificity in both samples ranged from 93.5-97.8%. CONCLUSIONS: Although STNPCR showed moderate sensitivity, the specificity is high; therefore, the test can be used as an auxiliary tool to diagnose TB in children. It is a rapid test that demonstrated better performance than other diagnostic tests in paucibacillary samples as it does in childhood tuberculosis.

The complement cascade in Alzheimer's disease: a systematic review and meta-analysis.

Genetic evidence implicates a causal role for the complement pathway in Alzheimer's disease (AD). Since studies have shown inconsistent differences in cerebrospinal fluid (CSF) and peripheral blood complement protein concentrations between AD patients and healthy elderly, this study sought to summarize the clinical data. Original peer-reviewed articles measuring CSF and/or blood concentrations of complement or complement regulator protein concentrations in AD and healthy elderly control (HC) groups were included. Of 2966 records identified, means and standard deviations from 86 studies were summarized as standardized mean differences (SMD) by random effects meta-analyses. In CSF, concentrations of clusterin (NAD/NHC = 625/577, SMD = 0.53, Z8 = 8.81, p < 0.005; I2 < 0.005%) and complement component 3 (C3; NAD/NHC = 299/522, SMD = 0.45, Z3 = 3.21, p < 0.005; I2 = 68.40%) were significantly higher in AD, but differences in C1q, C-reactive protein (CRP), serum amyloid protein (SAP), and factor H concentrations were not significant. In peripheral blood, concentrations of CRP were elevated in AD (NAD/NHC = 3404/3332, SMD = 0.44, Z43 = 3.43, p < 0.005; I2 = 93.81%), but differences between groups in C3, C4, C1-inhibitor, SAP, factor H and clusterin concentrations were not significant,and inconsistent between studies. Of 64 complement pathway proteins or regulators in the quantitative synthesis, trends in C1q, factor B, C4a, and late-stage complement pathway components (e.g. C9) in blood, C4 in CSF, and the membrane attack complex in blood and CSF, might be investigated further. The results collectively support elevated complement pathway activity in AD, which was best characterized by increased CSF clusterin concentrations and less consistently by CSF C3 concentrations. Complement activity related to anADdiagnosis was not reflected consistently by the peripheral blood proteins investigated.

A new hydrophilic polysulfone hemodialysis membrane can prevent platelet-neutrophil interactions and successive neutrophil activation.

Purpose:Microaggregates have often been observed during hemodialysis and are clearly associated with complications of hemodialysis therapy. In this study, we aimed to clarify the effects of two polysulfone membranes, with different abilities to activate blood cells, on the formation of these microaggregates; we also investigated their molecular mechanisms.Methods:Human whole blood was circulated through a mini-module dialyzer using the membranes in vitro; platelet–neutrophil complexes in blood were determined by flow cytometry. Isolated human neutrophils were incubated with the membranes in plasma, in the presence or absence of platelets, followed by flow cytometric analysis of intracellular reactive oxygen species and cell-surface activated CD11b on neutrophils.Results:CX-U, a conventional polysulfone membrane with remarkable cell activation, induced the formation of platelet–neutrophil complexes; however, NV-U, a new hydrophilic polysulfone membrane with slight or no cell activation, did not cause complex formation. Moreover, CX-U-induced reactive oxygen species production and the increase in activated CD11b expression on neutrophils were enhanced by platelets. On the other hand, NV-U hardly affected neutrophil activation, regardless of whether platelets were present or not. The enhancement of CX-U-induced neutrophil activations by platelets was greatly inhibited by anti-CD62P antibody.Conclusion:The ability of polysulfone membranes to activate blood cells is closely related to platelet–neutrophil interaction. Therefore, a biocompatible membrane, like NV-U, can be expected to prevent microaggregate formation during hemodialysis and avoid subsequent cell activation.

The biochemical characterization of a missense mutation m.8914C>T in ATP6 gene associated with mitochondrial encephalomyopathy

Mutations in ATP6 gene are frequent causes of mitochondrial encephalomyopathies. ATP6 gene encodes one subunit of complexⅤ. The present study described a missense mutation in ATP6 gene in a 8-year-old Chinese boy with mitochondrial encephalomyopathy. We identified one missense mutation in ATP6 gene (m.8914C>T) by mitochondrial DNA sequencing. This mutation altered the amino acid proline in serine, and alterative protein is predicted to be harmful. The mutation load in blood sample of patient is 59.49%. Activity of all mitochondrial complexes in blood are normal, however, the total function of mitochondrial oxidative phosphorylation were declined (including pathwayⅠ, pathwayⅡ and pathwayⅣ). The missense mutation (m.8914C>T) in ATP6 gene could result in abnormal function of complexV and is related with mitochondrial encephalomyopathy.

ОСОБЛИВОСТІ ІМУННОГО ГОМЕОСТАЗУ В ДИНАМІЦІ РОЗВИТКУ ЕКСПЕРИМЕНТАЛЬНОЇ БРОНХІАЛЬНОЇ АСТМИ

In this paper, it is shown that the modeling process of bronchial asthma (BA ) is accompanied by changes, in comparison with the control group, in the indicators of immune system response: T- &amp; B-lymphocytes, circulating immune complexes in blood of guinea pig males on the 4th, 18th, 25th day of experiment.The aim of the study – determination of some indices of immune system in blood of guinea pigs in the modeling process of BA on 4th, 18th, 25th day of the experiment.Materials and Methods. Experiments were conducted on 40 guinea pigs (males), with body weight 0.25–0.27 kg. Animals were divided into four groups of ten animals in each. Intact guinea pigs were the first group. Animals with experimental BA – the second, third, fourth group respectively on the 4th, 18th, 25th day of the experiment. Experimental BA was reproduced by V. I. Babych method. In blood of intact guinea pigs and animals with experimental BA , the number of T- and B-lymphocytes was determined by the method of E. F. Chernushenko, L. S. Kohosov, determination of the level of circulating immune complexes was carried out by the method of V. Haskova and co-authors. The results of the study were processed by the method of variation statistics using Student's criterion.Results and Discussion. The results of the studies showed unidirectional changes in individual parameters of the immune system, depending on the periods of the formation of BA : an increase in the number of B-lymphocytes and circulating immune complexes, a decrease in the number of T-lymphocytes for all of the studied days of the experiment.Conclusions. The obtained results indicate significant changes in the immune system parameters in the blood of experimental animals with BA and are important for understanding the pathogenesis of BA . These studies provide an opportunity to find the more perfect and effective methods of diagnosis of BA.

Efficient capture, rapid killing and ultrasensitive detection of bacteria by a nano-decorated multi-functional electrode sensor

In this work, we demonstrated a nano-decorated porous impedance electrode sensor for efficient capture, rapid killing and ultrasensitive detection of bacteria. The multi-functional sensor was prepared by a facile sonochemical method via in situ deposition of antibacterial prickly Zn-CuO nanoparticles and graphene oxide (GO) nanosheets on a Ni porous electrode. Due to the surface burr-like nanostructures, the nano-decorated impedance sensor exhibited very good bacterial-capture efficiency (70 − 80% in 20min) even at a low concentration of 50 CFU mL−1, rapid antibacterial rate (100% killing in 30min) and high detection sensitivity (as low as 10 CFU mL−1). More importantly, the nano-decorated sensor has proven to be highly effective in quantitative detection of bacteria in a biological sample, for example, a rat blood sample spiked with E. coli. Despite the complexity of blood, the sensor still exhibited excellent detection precision within 30min at bacteria concentrations ranging from 10 − 105 CFU mL−1. The simplicity, rapidity, sensitivity, practicability and multifunctionality of this impedance sensor would greatly facilitate applications in portable medical devices for on-the-spot diagnosis and even the possibility for simultaneous therapy of diseases caused by bacterial infections.

First study on iron complexes in blood and organ samples from thalassaemic and normal laboratory mice using Mössbauer spectroscopy.

Measurements of iron complexes and iron stores in the body are crucial for evaluation and management of chelation therapy targeted against iron accumulation or overload in blood and organs. In this work, blood and tissue samples from one normal and one thalassaemic laboratory mouse were studied using 57Fe Mössbauer spectroscopy at 78K for the first time. In contrast to human patients, these laboratory mice did not receive any medical treatment, thus the iron components present in the samples are not altered from their natural state. The Mössbauer spectra of blood, liver and spleen samples of the thalassaemic mouse were found to differ in shape and iron content compared with corresponding spectra of the normal mouse. These results demonstrate a basis for further exploitation of the thalassaemic mouse model to study thalassaemia and its treatment in more detail using Mössbauer spectroscopy.