Serum Fractions Research Articles

Karakterisasi Antibodi Poliklonal terhadap Aflatoksin M1

The aim of the research was characterized the polyclonal antibody at AFM for diagnostic reagens on track 1 and measure the levels of AFM in milk. The series of experiments are given a polyclonal antibody testing 1 dimetilaminobenzidin (DAB) by dot blot immunoassay (DBIA) so it will look brown color display on the positive control sample; purification and estimation antibody of AFM concentration with ammonium sulfate; 1 dialysis; fractionation using HiTrap Protein A HP column, measured using a spectrophotometer at a wavelength of 280 nm; and characterized of IgG using SDS-PAGE. DBIA test results showed a typical reaction betweenantigen AFM1-BSA with AFM1-BSA antibody to rabbit serum in the form of brown dots after addition of DAB substrate. The results of spectrophotometric against rabbit serum fractionation showed the type of IgG heavy chain.

Antioxidant defense system in familial hypercholesterolemia and the effects of lipoprotein apheresis

Oxidative stress is thought to play an important role in the pathogenesis of disorders associated with atherosclerosis. Alpha-tocopherol is considered to be an effective lipophilic antioxidant, which protects lipid membranes against peroxidation and thus prevents cell damage by reaction with free radicals. However, measurement of alpha-tocopherol concentration in serum does not reflect the content of α-tocopherol in membranes whereas erythrocyte alpha-tocopherol may be good indicator of antioxidative status. Therefore a simple isocratic reversed phase HPLC method has been developed and validated for the determination of alpha-tocopherol in human erythrocytes in a clinical setting. The content of alpha-tocopherol in human erythrocyte membrane and lipoperoxidation were studied in patients with severe hypercholesterolemia treated by lipoprotein apheresis. The group of hypercholesterolemic patients (n = 14) treated by lipoprotein apheresis was compared to healthy adult normolipidemic controls. After lipoprotein apheresis, the content of in membrane alpha-tocopherol did not change significantly despite decreased tocopherol in serum and lipoprotein fractions. We observed significantly decreased lipoperoxidation as revealed by serum TBARS, representing end products of lipid peroxidation, which increased from third day afterwards and remained significantly higher in comparison to controls until the next LDL-apheresis. We conclude that aggressive lipid lowering procedure with lipoprotein apheresis was associated with favorable transient decrease of lipoperoxidation. Simultaneously the cell membrane bound antioxidative defense mechanisms as reflected by the content of alpha-tocopherol in human erythrocyte membrane where not depressed in spite of its decreased plasma lipid carrier. Another variables involved remain to be investigated.

Effects of dietary supplementation with detoxified Rhus verniciflua sap on egg production, yolk lipid and intestinal microflora in laying hens

ObjectiveThis study was conducted to investigate the effects of dietary detoxified Rhus verniciflua sap (RVS) on production performance, egg quality, lipid fractions of egg yolk, liver and serum, and the profile of cecal microflora in laying hens.MethodsTwo hundred 52-week-old Hy-Line Brown layers were randomly divided into 4 groups with 5 replicates per group (2 hens per cage, 5 cages per replicate) and were provided with one of 4 experimental diets containing 0%, 0.05%, 0.1%, or 0.2% RVS, for 6 weeks. Due to unequal intervals of RVS doses, the interactive matrix language procedure of the SAS program was used to correct the contrast coefficients of orthogonal polynomials.ResultsThere were no differences in feed intake and egg weight among the groups. Egg production increased (linearly and quadratically, p<0.05) with increasing levels of RVS. Eggshell thickness increased (linear, p<0.05) as the level of RVS in diets increased. The levels of blood cholesterol and activities of glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase were not altered by dietary treatments. Increasing level of RVS increased (linear, p<0.05) the populations of cecal lactic acid bacteria. The content of yolk cholesterol decreased (linear, p< 0.05) with increasing levels of dietary RVS, although there were no significant differences in each lipid fraction of the liver.ConclusionThis study indicates that dietary RVS could improve laying performance and eggshell quality, and affect cecal lactic acid bacteria in a dose-dependent manner.

The effect of dietary fat on fatty acid composition, gene expression and vitamin status in pre-ruminant calves

Dietary saturated (SFA) and unsaturated fatty acids (UFA) alter fatty acid (FA) composition of various tissues, serum, and circulating immune cells. The objective of this study was to examine the effect of dietary SFA and UFA on adipose, liver, serum, polymorphonuclear (PMN) and peripheral blood mononuclear cells’ (PBMC) FA profiles, gene expression of selected inflammatory mediators, and their relation with serum lipid-soluble vitamin in pre-ruminant calves. Twelve Holstein male calves were randomly assigned to one of two treatments. Starting at 3 d of age, six calves received 120mL palm oil/day (SFA), and six calves received 80mL flaxseed oil plus 40mL conjugated linoleic acid/day (UFA). After 50 d, all animals were euthanized and samples were obtained. Fatty acid composition of non-esterified fatty acid (NEFA), neutral lipid (NL), and phospholipids (PL) was analyzed by gas-chromatography. High-performance liquid chromatography was used to detect α-tocopherol and retinol in liver, as well as α-tocopherol, retinol, and β-carotene in serum. In addition, liver and adipose tissue were analyzed for gene expression of interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12, interferon-γ, peroxisome proliferator-activated receptor-γ, TNF-α, retinol binding protein-4, and NF-κB. The PBMC were examined for IL-1β, IL-6, TNF-α and intercellular adhesion molecule-1; PMN cells were analyzed for caspase-1, IL-8 receptor, and l-selectin (l-SEL) expression. Data were analyzed using the Proc TTEST of SAS with significance declared at P≤0.05. Results showed that the UFA had greater α-linolenic acid compared to SFA in all three (NEFA, NL, and PL) lipid fractions of liver, adipose and serum, as well as PBMC and PMN. The greater content of α-LA in calves fed UFA resulted in greater eicosapentaenoic acid (EPA) in all three lipid fractions of serum, as well as NL and PL in adipose tissue. The UFA however, had lower γ-linolenic acid compared with SFA in all three lipid fractions of liver, as well as NL and PL in serum. Dietary UFA also increased total PUFA in all lipid fractions of serum and adipose. In addition, PBMC and PMN had greater EPA in calves fed UFA. Lipid-soluble vitamins in serum were reduced by dietary UFA. In contrast, UFA resulted in upregulation of l-SEL expression in PMN. This may indicate that UFA elevated the substrate for PUFA biosynthesis, but possibly degraded lipid-soluble vitamins to protect these FA from oxidation. Greater circulating PUFA may influence the migration of PMN from the blood to tissues, affecting overall inflammatory responses.

Impact of Neuro-Behçet Disease Immunoglobulin G on Neuronal Apoptosis.

Parenchymal neuro-Behçet disease (NBD) is encountered in 5%-15% of Behçet disease (BD) patients and is characterized by inflammation of the brainstem and diencephalon structures. Neuronal apoptosis has been shown to participate in neuronal cell loss. Anti-neuronal antibodies have been identified in NBD patients. However, the pathogenic properties of these antibodies have not been studied. To delineate the potential pathogenic activity of serum antibodies on neurons, pooled sera from seven NBD patients and seven healthy controls were divided into purified immunoglobulin G (IgG) and IgG-depleted serum fractions, and each fraction was administered to cultured SH-SY5Y neuroblastoma cells. Cell death was evaluated with a toxicity assay and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Moreover, expression levels of several apoptosis markers were evaluated with real time polymerase chain reaction (PCR). Administration of NBD IgG to cultured SH-SY5Y cells induced significantly increased cell death and apoptosis compared with other treatments. NBD IgG also enhanced the mRNA expression levels of major apoptosis and cell survival pathway factors. Our results suggest that IgGs isolated from the sera of NBD patients have a neurotoxic activity that is presumably mediated by apoptotic mechanisms.

Evidence for host genetic regulation of altered lipid metabolism in experimental toxoplasmosis supported with gene data mining results.

Toxoplasma gondii is one of the most successful parasites on Earth, infecting a wide array of mammals including one third of the global human population. The obligate intracellular protozoon is not capable of synthesizing cholesterol (Chl), and thus depends on uptake of host Chl for its own development. To explore the genetic regulation of previously observed lipid metabolism alterations during acute murine T. gondii infection, we here assessed total Chl and its fractions in serum and selected tissues at the pathophysiological and molecular level, and integrated the observed gene expression of selected molecules relevant for Chl metabolism, including its biosynthetic and export KEGG pathways, with the results of published transcriptomes obtained in similar murine models of T. gondii infection. The serum lipid status as well as the transcript levels of relevant genes in the brain and the liver were assessed in experimental models of acute and chronic toxoplasmosis in wild-type mice. The results showed that acute infection was associated with a decrease in Chl content in both the liver and periphery (brain, peripheral lymphocytes), and a decrease in Chl reverse transport. In contrast, in chronic infection, a return to normal levels of Chl metabolism has been noted. These changes corresponded to the brain and liver gene expression results as well as to data obtained via mining. We propose that the observed changes in Chl metabolism are part of the host defense response. Further insight into the lipid metabolism in T. gondii infection may provide novel targets for therapeutic agents.

Mass Spectrometry Analysis of Chromium-Binding Low-Molecular-Weight Serum Fractions

The current work is focused on the characterization of the chromium(III)-containing low-molecular-weight (LMW) fractions of blood serum by mass spectrometry. The LMW fractions of serum binding-Cr(III) were collected by organic solvent precipitation using acetonitrile. The chromium(III) compounds used were [Cr(pic) 3 ], where pic = 2-pyridinecarboxylato(-) or A , and the Cr(III) propionate complex, [Cr 3 O(OCOEt) 6 (OH 2 ) 3 ]) + or B . The new signals appeared in the mass spectra in the presence of added Cr(III) compounds, either A or B . This indicated that common LMW Cr containing serum fractions were formed on addition of Cr(III) to serum. The new signals contained three distinct peaks with a shift of 16, corresponding to ions of m/z +1083, +1099, and +1115 Da, which consisted of 4 Cr atoms with bridging ligands.

Abstract 192: Intra-Individual Temporal Variation of Lipid and Apolipoprotein Levels in Patients With Type II Diabetes Mellitus and Normolipidemic Individuals

Hypertriglyceridemia is frequently observed with diabetes mellitus (DM) and is associated with increased cardiovascular disease (CVD) risk. In conditions of insulin deficiency or resistance, increased levels of free fatty acids are seen by the liver, which trigger increased lipogenesis and secretion of triglyceride (TG) rich lipoprotein (TRL) particles. However, a high number of specific ApoC-III on TRL particles suppress their return to the liver leading to a continuous circulation of highly atherogenic TRL remnants. To better understand the effect of intra-individual temporal variations on the differentiation of lipid and apolipoprotein profiles in DM vs. normal individuals, a small time course study over a 4 week period was performed on 3 normolipidemic, and 3 type II DM patients. Our laboratory has developed a method to separate lipoproteins based on hydrodynamic size coupled with mass spectrometry based analysis. Whole serum and lipoprotein size fractions were analyzed for 8 apolipoproteins (ApoA-I, ApoA-II, ApoA-IV, B-100, ApoC-I, ApoC-II, ApoC-III and E) and non-polar lipids (FC, CE, TG) while monitoring other proteins (ApoD, ApoM, CETP, LCAT, PLTP, PON1 and SAA4). In whole serum over the course of 4 weeks, ApoA-IV levels of DM vs. normal subjects differed the most, 3.2 (0.7) vs. 1.8 (0.4) μM, respectively, more significantly than intra-group or intra-individual variations. On the contrary, significantly lower whole serum levels were found for FC, CE, ApoM and PON1 (Prob&lt;0.01). HDL-ApoC-III/LDL-ApoC-III was also lower in DM vs. normal subjects, 3.0 (1.9) vs. 7.2 (4.1). By metrics of molar ratio measured in 20-35 nm LDL fractions, the most significant difference was found between DM vs. normal subjects in ApoC-III/ApoB-100 (1.8 (0.6) vs. 0.9 (0.3)), C-II/ApoB-100 (0.8 (0.4) vs. 0.4 (0.2)) and ApoE/ApoC-III (0.11 (0.09) vs. 0.24 (0.15)). These LDL composition differences indicate lower lipase activity and inhibition of LDL uptake. However, no significant differences were observed in these molar ratios as function of LDL particle size. In conclusion, due to less significant temporal deviations, apolipoprotein molar ratios are a more useful and informative measure of CVD risk in DM subjects than LDL size distribution characteristics.

Abstract 194: Lipid and Apolipoprotein Size Profile Differences Between Low and High Body Mass Index Individuals in Response to Fat Challenge

Clearance of elevated triglycerides (TG) level in response to fat challenge is a clinically significant indicator of CVD risk. HDL can play a key role in facilitating TG metabolism explaining the reverse correlation of HDL levels with CVD risk. In this study 6 individuals, low BMI (21-25) and 3 high BMI (30-49) were challenged with a meal containing ~90 g of total fat. Blood samples were drawn in fasting state before meal and 3 times after meal in 2, 4 and 6 hours. Serum was analyzed by a quantitative, multiplexed analytical workflow which included analysis of whole serum and the separation of lipoproteins by asymmetric flow field-flow fractionation where fractions were collected with 1 nm increments of 7-15 nm (HDL), 20-30 nm (LDL) and &gt;30 nm lipoprotein classes. Dynamic light scattering was used to determine hydrodynamic size in each size fraction. Whole serum and individual fractions were analyzed to determine concentrations of apolipoproteins (apo A-I, A-II, A-IV, B, C-I, C-II, C-III and E), and non-polar lipids (FC, CE and TG), using two parallel liquid chromatography tandem mass spectrometry methods developed in our laboratory. In whole serum, the total TG levels for low and high BMI donors, relative to pre-meal levels (57-137 mg/dL and 111-183 md/dL), peaked at between the 2 and 4 hours post meal (103-214 mg/dL and 210-309 mg/dL). LCAT were lower (20%) and CETP levels were higher (30%), in low vs high BMI groups, but did not show change in repose to the fat challenge. In both low and high BMI groups, the most significant change between pre-meal and post-meal after 2 hours occurred in HDL-ApoC-I, HDL-ApoC-II, and HDL-ApoC-III levels mainly in the HDL size range of 9-12 nm (medium HDL) and at 13-16 nm (large HDL), with concurrent TG increase observed in LDL and remnant fractions (Prob-

Dissecting the polyclonal antibody response of nurse sharks (Ginglymostoma cirratum) to immunization with influenza A virus suggests different roles for IgNAR and IgM, but a common immunodominance hierarchy with hemagglutinin on top

Abstract Cartilaginous fishes are the earliest extant phylogenic group to harness germline recombination and somatic hypermutation of immunoglobulins antibodies (Igs) for immunity. In this study we immunized and 1-year-later boosted, two adult nurse sharks (Ginglymostoma cirratum) with inactivated H1 influenza A virus (IAV). After immunization, polyclonal sera from these sharks had strong antiviral properties. Primary immune and recall sera neutralized viral growth in cell culture, caused hemagglutination inhibition, and inhibited neuraminidase (NA) activity. By ELISA and protein microarray, we examined the contribution of two shark Ig isotypes, IgNAR and IgM. Unique to cartilaginous fishes, IgNAR are heavy-chain-only Igs. IgNAR from immunized sharks bound only the immunizing strain of virus. But, flow cytometry of spleen and blood revealed ~1% of IgNAR-positive cells bound both H1 and H5 HA subtypes. While these crossreactive cells exist, any crossreactive antibodies made by cells in these lineages were below our limit of detection. By contrast, both preimmune and immune serum IgM was highly crossreactive, binding H1, H3, H5, and even headless HA. In cartilaginous fish, secreted IgM comes in two independently-produced forms high affinity monomeric (7s) and pentameric (19s). Serum fractionation determined these crossreactive IgM antibodies were 19s. While capable of binding virus, we found 19s alone could not neutralize virus. Despite these differences, regardless of isotype, and in-line with our previous work with mice and lamprey, the immunodominance hierarchy of shark vaccine-induced IgNAR and IgM antibodies to IAV is consistent—HA is dominant, followed by NA, and then core proteins, such as, nucleoprotein and matrix.

Algal supplementation of vegetarian eating patterns improves plasma and serum docosahexaenoic acid concentrations and omega-3 indices: a systematic literature review.

Vegetarians are likely to have lower intakes of preformed docosahexaenoic acid (DHA) than omnivorous populations who consume fish and animal products. As such, vegetarian populations have omega-3 indices up to 60% lower than those who consume marine products. Algae, the primary producer of DHA in the marine food chain, offer an alternative source of DHA for those who do not consume marine or animal products. This systematic review aims to examine the evidence for the relationship between supplementation with algal forms of DHA and increased DHA concentrations in vegetarian populations. The SCOPUS, Science Direct and Web of Science scientific databases were searched to identify relevant studies assessing the effect of algal DHA consumption by vegetarian (including vegan) populations. Four randomised controlled trials and two prospective cohort studies met the inclusion criteria. All included studies reported algal sources of DHA significantly improve DHA concentrations (including plasma, serum, platelet and red blood cell fractions), as well as omega-3 indices, in vegetarian populations. An evident time or dose response was not apparent given the small number of studies to date. Future studies should address long chain n-3 polyunsaturated fatty acid deficiencies in vegetarian populations using algal DHA and explore the potential physiological and health improvements in these individuals.

A serum and platelet-rich plasma serotonin assay using liquid chromatography tandem mass spectrometry for monitoring of neuroendocrine tumor patients

BackgroundSerotonin is used for the diagnosis and follow-up of neuroendocrine tumors (NET). We describe the analytical and clinical validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) based serotonin assay for serum and platelet-rich plasma (PRP). MethodsAn LC-MS/MS based method for serum and PRP serotonin was validated by determination of assay imprecision, carry-over, linearity, interference, recovery, sample stability and a matrix/method comparison of serum and PRP serotonin was made with whole blood serotonin. Furthermore, upper limits of normal were determined and serotonin concentrations of healthy individuals, 14 NET patients without evidence of disease and 51 NET patients with evidence of disease were compared. ResultsFor serum and PRP fractions, total assay imprecision was <5%. All correlation coefficients were 0.98 and the serum and platelet-rich serotonin upper limit of normal were 5.5nmol/109 platelet and 5.1nmol/109 platelet, respectively. NET patients with confirmed evidence of disease had significantly higher serum and PRP serotonin levels when compared to NET patients without evidence of disease and healthy volunteers. ConclusionsLC-MS/MS based serum and PRP serotonin assays were developed with suitable analytical characteristics. Furthermore, serum and PRP serotonin was found to be useful for monitoring NET patients.

MP06-02 SERUM SIALYL HYBRID TYPED N-GLYCAN LEVELS PREDICTS EARLY ABMR IN LIVING DONOR KIDNEY TRANSPLANT PATIENTS

You have accessJournal of UrologyTransplantation & Vascular Surgery: Renal Transplantation & Vascular Surgery I1 Apr 2017MP06-02 SERUM SIALYL HYBRID TYPED N-GLYCAN LEVELS PREDICTS EARLY ABMR IN LIVING DONOR KIDNEY TRANSPLANT PATIENTS Daisuke Noro, Tohru Yoneyama, Shingo Hatakeyama, Yuki Tobisawa, Kazuyuki Mori, Yasuhiro Hashimoto, Takuya Koie, Masakazu Tanaka, Shinichiro Nishimura, Hideo Sasaki, Mitsuru Saito, Hiroshi Harada, Tatsuya Chikaraishi, Hideki Ishida, Kazunari Tanabe, Shigeru Satoh, and Chikara Ohyama Daisuke NoroDaisuke Noro More articles by this author , Tohru YoneyamaTohru Yoneyama More articles by this author , Shingo HatakeyamaShingo Hatakeyama More articles by this author , Yuki TobisawaYuki Tobisawa More articles by this author , Kazuyuki MoriKazuyuki Mori More articles by this author , Yasuhiro HashimotoYasuhiro Hashimoto More articles by this author , Takuya KoieTakuya Koie More articles by this author , Masakazu TanakaMasakazu Tanaka More articles by this author , Shinichiro NishimuraShinichiro Nishimura More articles by this author , Hideo SasakiHideo Sasaki More articles by this author , Mitsuru SaitoMitsuru Saito More articles by this author , Hiroshi HaradaHiroshi Harada More articles by this author , Tatsuya ChikaraishiTatsuya Chikaraishi More articles by this author , Hideki IshidaHideki Ishida More articles by this author , Kazunari TanabeKazunari Tanabe More articles by this author , Shigeru SatohShigeru Satoh More articles by this author , and Chikara OhyamaChikara Ohyama More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.247AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES ABMR is a diagnostic challenge in living donor kidney transplant (LKTx) medicine, and there is a need to identify predictive markers of ABMR to improve graft survival. The use of serum N-glycans as a predictive biomarker of ABMR has not yet been tested. In the present study, we performed serum N-glycomics in transplant patients and evaluated its potential as a predictive serum-based biomarker of early ABMR. METHODS N-glycomics in whole serum and immunoglobulins (Igs) fraction were performed in randomly selected 16 recipients with biopsy-proven ABMR occurred within 1 month after LKTx, 40 recipients with biopsy-proven TCMR, and 141 recipients without any adverse events. The putative structure of N-glycans was analyzed by MALDI-TOF-MS analysis. RESULTS Serum sialyl hybrid-type N-glycans (m/z 1709, 1871, and 2033) levels before LKTx and on postoperative Day 1 (POD1) was significantly lower in recipients who developed ABMR than non-ABMR group. The m/z 2033 N-glycan <1.3 μM and the presence of preformed donor-specific antibodies (DSA) on POD1 were found to yield a higher odds ratio for prediction of ABMR than did other factors according to logistic regression analysis. Receiver-operating characteristic area under the curve for m/z 2033 < 1.3 μM combined with preformed-DSA status was 0.86. Combined preformed DSA with m/z 2033 N-glycan status; thus, double-positive patients (preformed-DSA positive and m/z 2033 < 1.3 μM) or single-positive patients (preformed-DSA positive or negative and m/z 2033 > 1.3 μM or < 1.3 μM) can cover all 16 ABMR cases. These results suggest that the combined indicator holds promise for identification of patients who will not develop ABMR by means of serum samples collected on POD1. Although, N-glycan profile of immunoglobulin (Ig)s fractions compared with those of whole-serum, ABMR-related N-glycans in Igs fractions were not detected. Therefore, the ABMR-related N-glycans carrying proteins are not an Igs. CONCLUSIONS The serum m/z 2033 sialyl hybrid-type N-glycan combined with preformed DSA status may predict acute ABMR in patients undergoing LKTx. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e72-e73 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Daisuke Noro More articles by this author Tohru Yoneyama More articles by this author Shingo Hatakeyama More articles by this author Yuki Tobisawa More articles by this author Kazuyuki Mori More articles by this author Yasuhiro Hashimoto More articles by this author Takuya Koie More articles by this author Masakazu Tanaka More articles by this author Shinichiro Nishimura More articles by this author Hideo Sasaki More articles by this author Mitsuru Saito More articles by this author Hiroshi Harada More articles by this author Tatsuya Chikaraishi More articles by this author Hideki Ishida More articles by this author Kazunari Tanabe More articles by this author Shigeru Satoh More articles by this author Chikara Ohyama More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

A Comparative Biochemical Study of Proteins Profile in Iraqi Children and Adolescent with ?-Thalassemia

The aim of the present research is to study different protein fractions in sera of children and adolescent with β –thalassemia major and minor and to compare the results with that of healthy control.One hundred fifty children and adolescents were enrolled in this study,including 50 patients with β- thalassemia major , 50 patients with β- thalassemia minor as pathological control group and another apparently 50 healthy individuals as a control group. The age of all studied groups ranged from (4-18)years.Total protein, albumin and immunoglobulins were estimated in sera of all subjects. A Significant decrease was found in the total protein and albumin levels in sera of both major and minor thalassemic patients compared to normal groups. A Significant increase in immunoglobulin levels (IgG, IgM and IgA) was found in the sera of major and minor β-thalassemia patients compared to control.Different protein parts in sera of all subjects were detected using cellulose acetate electrophoresis.The results revealed significant reduction in β- globulin fractions in β- thalassemia major patients compared to the normal and pathological control groups. Significant elevations in γ- globulins fractions were observed in both major β- thalassemia and minor β- thalassemia as compared to normal control group. As a Conclusion the alteration in some protein parts occurred which is more obvious in major thalassemia patients compared to the normal and pathological control groups.&#x0D; Key words: Protein Parts , Elecrtrophoresis , β-Thalassemia

Evaluation of lipid fractions in serum of premenopausal and postmenopausal women in Kirkuk city

Evaluation of lipid fractions in serum of premenopausal and postmenopausal women in Kirkuk city

Clinical implications of serum N-glycan profiling as a diagnostic and prognostic biomarker in germ-cell tumors.

Serum biomarker monitoring is essential for management of germ‐cell tumors (GCT). However, not all GCT are positive for conventional tumor markers. We examined whether serum N‐glycan‐based biomarkers can be applied for detection and prognosis in patients with GCT. We performed a comprehensive N‐glycan structural analysis of sera from 54 untreated GCT patients and 103 age‐adjusted healthy volunteers using glycoblotting methods and mass spectrometry. Candidate N‐glycans were selected from those with the highest association; cutoff concentration values were established, and an N‐glycan score was created based on the number of positive N‐glycans present. The validity of this score for diagnosis and prognosis was analyzed using a receiver operating characteristic (ROC) curve. We identified five candidate N‐glycans significantly associated with GCT patients. The accuracy of the N‐glycan score for GCT was significant with an area‐under‐the‐curve (AUC) value of 0.87. Diagnostically, the N‐glycan score detected 10 of 12 (83%) patients with negative conventional tumor markers. Prognostically, the N‐glycan score comprised four candidate N‐glycans. The predictive value of the prognostic N‐glycan score was significant, with an AUC value of 0.89. A high value prognostic N‐glycan score was significantly associated with poor prognosis. Finally, to identify a potential carrier protein, immunoglobulin (Ig) fractions of sera were subjected to N‐glycan analysis and compared to whole sera. Candidate N‐glycans in Ig‐fractions were significantly decreased; therefore, the carrier protein for candidate N‐glycans is likely not an immunoglobulin. In summary, our newly developed N‐glycan score seems to be a practical diagnostic and prognostic method for GCT.

PreImplantation factor (PIF) protects cultured embryos against oxidative stress: relevance for recurrent pregnancy loss (RPL) therapy.

Recurrent pregnancy loss (RPL) affects 2-3% of couples. Despite a detailed work-up, the etiology is frequently undefined, leading to non-targeted therapy. Viable embryos and placentae express PreImplantation Factor (PIF). Maternal circulating PIF regulates systemic immunity and reduces circulating natural killer cells cytotoxicity in RPL patients. PIF promotes singly cultured embryos’ development while anti-PIF antibody abrogates it. RPL serum induced embryo toxicity is negated by PIF. We report that PIF rescues delayed embryo development caused by <3 kDa RPL serum fraction likely by reducing reactive oxygen species (ROS). We reveal that protein disulfide isomerase/thioredoxin (PDI/TRX) is a prime PIF target in the embryo, rendering it an important ROS scavenger. The 16F16-PDI/TRX inhibitor drastically reduced blastocyst development while exogenous PIF increased >2 fold the number of embryos reaching the blastocyst stage. Mechanistically, PDI-inhibitor preferentially binds covalently to oxidized PDI over its reduced form where PIF avidly binds. PIF by targeting PDI/TRX at a distinct site limits the inhibitor’s pro-oxidative effects. The >3kDa RPL serum increased embryo demise by three-fold, an effect negated by PIF. However, embryo toxicity was not associated with the presence of putative anti-PIF antibodies. Collectively, PIF protects cultured embryos both against ROS, and higher molecular weight toxins. Using PIF for optimizing in vitro fertilization embryos development and reducing RPL is warranted.

Quantifying glucose and lipid components in human serum by Raman spectroscopy and multivariate statistics.

Raman spectroscopy has been employed in the quantitative analysis of biochemical components in human serum. This study aimed to develop a spectral model to estimate the concentration of glucose and lipid fractions in human serum, thus evaluating the feasibility of Raman spectroscopy technique for diagnostic purposes. A total of 44 samples of blood serum were collected from volunteers submitted to routine blood biochemical assay analysis. The biochemical concentrations of glucose, triglycerides, cholesterol, and high-density and low-density lipoproteins (HDL and LDL) were obtained by colorimetric method. Serum samples (200μL) were submitted to Raman spectroscopy (830nm, 250mW, 50-s accumulation). The spectra of sera present peaks related to the main constituents, particularly proteins and lipids. A quantitative model based on partial least squares (PLS) regression has been developed to estimate the concentration of these compounds, taking the biochemical concentrations assayed by the colorimetric method as sample's actual concentrations. The PLS model based on leave-one-out cross-validation approach estimated the concentration of triglycerides and cholesterol with r = 0.98 and 0.96, and root mean square error of 35.4 and 15.9mg/dL, respectively. For the other biochemicals, the r was ranging from 0.75 to 0.86. These results evidenced the possibility of performing biochemical assay in blood serum samples by Raman spectroscopy and PLS regression and may be employed as a means of diagnosis in routine clinical analysis.

Effects of in vivo and in vitro administration of neuro-Behcet’s disease IgG

Antibodies directed against membrane antigens of neuronal axonal processes (neuropil) have been recently identified in neuro-Behcet's disease (NBD) patients. To delineate the potential pathogenic action of these antibodies, pooled sera from seven NBD patients with neuropil antibodies and seven healthy controls were divided into purified IgG and IgG-depleted serum (IgG-DS) fractions and each fraction was administered into lateral ventricles of rats. NBD IgG-injected rats showed reduced locomotor activity in the open field test as compared to NBD IgG-DS, healthy control IgG, healthy control IgG-DS and PBS injected rats (n=10 for each group). There were no significant differences among treatment groups by means of anxiety-like behaviors (assessed by elevated plus maze test) and learning/memory functions (assessed by passive avoidance test). Administration of NBD IgG on cultured SH-SY5Y neuroblastoma cells induced significantly increased cell death and apoptosis (as measured by nucleosome levels in the supernatants) as compared to other treatment groups. Our results suggest that IgGs isolated from sera of neuropil antibody-positive NBD patients have a neurotoxic action, which is presumably mediated by apoptotic mechanisms. Motor deficits frequently observed in NBD patients might at least partially be caused by the pathogenic action of anti-neuronal IgG.

Green and roasted coffee extracts as antioxidants in \u03b2TC3 cells with induced oxidative stress and lipid accumulation inhibitors in 3T3L1 cells, and their bioactivity in rats fed high fat diet

Chlorogenic acids from coffee show high antioxidant activity that contributes to inhibition of oxidative stress, which results in anticancer, antidiabetic and antiobesity coffee activities. Coffee roasting causes their partial decomposition and partial incorporation into Maillard reaction products also of high antioxidant activity, and it is still not clear whether more favorable impact on health is associated with the consumption of green or roasted coffee. The aim of the study was to evaluate the impact of coffee roasting process and also of the degree of chlorogenic acid esterification on bioactivity of coffee extracts. Extracts of green and roasted coffee and their fractions containing mono- and dichlorogenic acids (six extracts) were obtained by purification and optional fractionation of crude extracts using preparative centrifugal partition chromatography technique. The extracts were evaluated in vitro in terms of scavenging capacity, influence on lipids accumulation in 3T3L1 adipocytes, cytoprotective potential decreasing oxidative stress induced by cellular pro-oxidants in βTC3 cells, and in vivo by selected physiological indices in rats fed diets supplemented with the extracts. Roasted coffee extracts, in particular hydrophilic fraction containing monochlorogenic acids and Maillard reaction products, showed the highest scavenging capacity and contributed to the highest level of antioxidants in the hydrophilic fraction of rats serum and may be potentially used as anticancer nutraceutical with oxidative stress inhibitor activity. Non-fractionated green coffee extract exhibited antidiabetic activities including inhibition of adipogenesis, while in vivo it significantly lowered the levels of triglycerides, glucose and oxidized glutathione.