P1077PERSISTENCE OF CIRCULATING RESIDUAL HEPARIN IN ESRD PATIENTS UNDERGOING MAINTENANCE HEMODIALYSIS

Abstract Background and Aims ESRD patients who receive routine maintenance hemodialysis are administered with unfractionated heparin to prevent thrombotic complications. The hemostatic dysregulation along with detectable levels of circulating heparin may cause them to be in a hypocoagulable state. The purpose of this study is to determine the circulating levels of heparin in ESRD patients and its characterization using heparinase digestion methods. Method Blood plasma samples collected at routine pre-dialysis sessions from 95 ESRD patients undergoing maintenance hemodialysis were analyzed for the presence of residual heparin utilizing standard laboratory methods such as aPTT, Anti-Xa and Anti-IIa activities On a centrifugal analyzer (ACL-Elite: Instrumentation Laboratory, Bedford, MA). The levels of heparin were calculated in terms of units per ml relative to the USP reference standard. Heparinase digestion was used to confirm the presence of heparin. Results Blood plasma samples collected at routine pre-dialysis sessions from 95 ESRD patients undergoing maintenance hemodialysis were analyzed for the presence of residual heparin utilizing standard laboratory methods such as aPTT, Anti-Xa and Anti-IIa activities On a centrifugal analyzer (ACL-Elite: Instrumentation Laboratory, Bedford, MA). The levels of heparin were calculated in terms of units per ml relative to the USP reference standard. Heparinase digestion was used to confirm the presence of heparin. Conclusion The presence of residual heparin was demonstrated by both clot- based and amidolytic assays in the plasma samples collected from ESRD patients prior to their next-dialysis session. Since these samples were obtained 3 days following the last dialysis session the presence of significant levels of heparin was surprising. Upon heparinase treatment of these samples, the aPTT and the anti-Xa and IIa tests were restored to near normal levels. Our studies confirm the presence of residual heparin in pre-dialysis plasma samples obtained from ESRD patients. The Anti-IIa activity was greater pre-heparinase and it was not decreased to the same extent as Anti-Xa after heparinase digestion. These results suggest that heparin found in ESRD patient’s plasma is of high molecular weight origin with delayed clearance.

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Role of Platelet Activation in the Pathogenesis of Heart Failure in End-Stage Renal Disease Patients

Blood ◽

10.1182/blood.v126.23.4636.4636 ◽

2015 ◽

Vol 126 (23) ◽

pp. 4636-4636

Author(s):

Ryan McMillan ◽

Vinod Bansal ◽

Leonidas Skiadopoulos ◽

Debra Hoppensteadt ◽

Omer Iqbal ◽

...

Keyword(s):

Hemodialysis Patients ◽

Maintenance Hemodialysis ◽

Dialysis Session ◽

Positive Correlation ◽

Activated Platelets ◽

Stage Renal Disease ◽

End Stage ◽

Esrd Patients ◽

Maintenance Hemodialysis Patients

Abstract Introduction: Heart failure (HF) is prevalent in patients with End-Stage Renal Disease (ESRD) with an incidence of approximately 40%. ESRD patients exhibit both quantitative and qualitative changes in platelets due to the uremic milleu and / or stress caused by the dialysis filtration membranes. Platelet Factor 4 (PF4) and Platelet Derived Growth Factor (PDGF) are released from the alpha granules of activated platelets and play a role in heparin neutralization and angiogenesis, respectively. The purpose of this study was to determine the interplay of platelets and their released mediators and their relevance to the pathogenesis of HF in ESRD patients. Material and Methods: Under IRB approval, ninety blood samples from maintenance hemodialysis patients at Loyola University outpatient dialysis unit were collected prior to dialysis session and stored at -70°C. Twenty-five male and twenty-five female plasma samples from healthy individuals were obtained as a control (George King Biomedical Overland Park, KS). Maintenance hemodialysis patients' and healthy volunteers' plasma samples were used to profile PDGF-BB, Heparin anti Xa, and PF4 using commercial sandwich and competitive ELISA kits (R&D Systems, Minneapolis, MN | Hyphen Biomed, Neuville-sur-oise, France). In addition, patients' HF diagnoses, comorbidities, medications, and clinical laboratory parameters were reviewed through the patients' medical records. Results: Plasma biomarkers PF4 (P < 0.0001; % change = 247.76) and heparin level as measured by an anti Xa methods were significantly increased (P < 0.05) in patients with ESRD in comparison to the values obtained in normal healthy volunteers. PDGF-BB was not found to be significantly increased (P = 0.4045 ; % change = 40.3%). PF4, Heparin levels, and PDGF-BB were not found to be significantly elevated in ESRD (+) HF vs ESRD (-) HF (P > 0). In male ESRD (+) CHF vs ESRD (-) HF, platelet counts were significantly decreased (P = 0.0476 ; % change = -16.5). Systolic BP and PLT count had a positive correlation in both male and female ESRD patients and in male ESRD (+) HF patients (P = 0.016 and P = 0.044, respectively), but not in female ESRD HF (+) patients (P = 0.172). PLT count and PDGF were found to have a positive correlation in ESRD HF (+) patients (P = 0.045). In ESRD (+) HF patients, PF4 had a positive correlation with proBNP (P = 0.045) and Heparin levels had a negative correlation with proBNP (P = 0.045). Discussion: Elevated PF4 in the ESRD patients compared to normals suggest that there may be increased platelet granule release or an upregulation of PF4. The platelets may be activated due to dialysis procedure stress from the previous session, the uremic environment, or heparin-PF4 complexed with antibodies binding to the platelet surface. Elevated heparin levels in the ESRD patients compared to normal suggests that heparinization at dialysis session may lead to an anticoagulated state between sessions in several of these patients. Since these patients are continually heparinized, it is expected to see increased levels of platelet release products such as PF4 due to their mobilization from platelets. Activated platelets may lead to platelet exhaustion and thrombocytopenic responses. Interestingly thrombocytopenia is only noticed in male ESRD (+) HF patients suggesting gender differences and the potential role of hormonal regulation. These results suggest that both the quantitative and qualitative defects in platelets play an important role in the mediation of the pathogenesis of HF in ESRD patients underscoring the potential benefit of antiplatelet drugs. Disclosures No relevant conflicts of interest to declare.

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Alterations in lipid profile upon uterine fibroids and its recurrence

Scientific Reports ◽

10.1038/s41598-021-89859-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Narine M. Tonoyan ◽

Vitaliy V. Chagovets ◽

Natalia L. Starodubtseva ◽

Alisa O. Tokareva ◽

Konstantin Chingin ◽

...

Keyword(s):

Mass Spectrometry ◽

Blood Plasma ◽

Lipid Profile ◽

Plasma Levels ◽

Uterine Fibroids ◽

Control Group ◽

Cholesterol Esters ◽

Plasma Samples ◽

Diagnostic Models ◽

First Time

AbstractUterine fibroids (UF) is the most common (about 70% cases) type of gynecological disease, with the recurrence rate varying from 11 to 40%. Because UF has no distinct symptomatology and is often asymptomatic, the specific and sensitive diagnosis of UF as well as the assessment for the probability of UF recurrence pose considerable challenge. The aim of this study was to characterize alterations in the lipid profile of tissues associated with the first-time diagnosed UF and recurrent uterine fibroids (RUF) and to explore the potential of mass spectrometry (MS) lipidomics analysis of blood plasma samples for the sensitive and specific determination of UF and RUF with low invasiveness of analysis. MS analysis of lipid levels in the myometrium tissues, fibroids tissues and blood plasma samples was carried out on 66 patients, including 35 patients with first-time diagnosed UF and 31 patients with RUF. The control group consisted of 15 patients who underwent surgical treatment for the intrauterine septum. Fibroids and myometrium tissue samples were analyzed using direct MS approach. Blood plasma samples were analyzed using high performance liquid chromatography hyphened with mass spectrometry (HPLC/MS). MS data were processed by discriminant analysis with projection into latent structures (OPLS-DA). Significant differences were found between the first-time UF, RUF and control group in the levels of lipids involved in the metabolism of glycerophospholipids, sphingolipids, lipids with an ether bond, triglycerides and fatty acids. Significant differences between the control group and the groups with UF and RUF were found in the blood plasma levels of cholesterol esters, triacylglycerols, (lyso) phosphatidylcholines and sphingomyelins. Significant differences between the UF and RUF groups were found in the blood plasma levels of cholesterol esters, phosphotidylcholines, sphingomyelins and triacylglycerols. Diagnostic models based on the selected differential lipids using logistic regression showed sensitivity and specificity of 88% and 86% for the diagnosis of first-time UF and 95% and 79% for RUF, accordingly. This study confirms the involvement of lipids in the pathogenesis of uterine fibroids. A diagnostically significant panel of differential lipid species has been identified for the diagnosis of UF and RUF by low-invasive blood plasma analysis. The developed diagnostic models demonstrated high potential for clinical use and further research in this direction.

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The Complex Biochemical Study of the Blood Plasma, Obtained by Manual Plasmapheresis

Family Medicine ◽

10.30841/2307-5112.5.2016.248853 ◽

2016 ◽

pp. 155-157

Author(s):

Andrii Korzh

Keyword(s):

Blood Plasma ◽

Biochemical Parameters ◽

Biochemical Study ◽

Plasma Samples ◽

Fluorometric Method ◽

Plasma Donation ◽

Individual Index ◽

First Time ◽

Age And Sex ◽

Donor Plasma

The plasma samples of 34 primary donors (22 men and 12 women) for the first time given the plasma by automated plasmapheresis (control surveillance), and 54 active donors of blood plasma (40 men and 14 women) being donors with non-less 14 days interval between donations, have been examined. The active male donors’ plasma averaged at 18,63±1,71 with individual index fluctuations from 2 to 78, female donors’ – 14,09±1,95 with individual index fluctuations from 2 to 45. The method of plasma obtaining is a manual plasmapheresis method. The surveyed groups were homogeneous for age and sex. Hematologic and biochemical parameters of all those persons have been examined and, basing on the conclusion of the professionals, everyone was admitted to the plasma donation. The content of middle mass molecules in plasma were determined by method of N.I. Gabrieljan, V.I. Lipatovoj (1984). The content of biogenic amines free fractions in plasma were determined by fluorometric method of B.V. Mikhailichenko, S.V. Vydyborets (1999). Analysis of the results showed that in the donor plasma samples obtained by manual plasmapheresis level of middle mass molecules, histamine, serotonin is significantly higher. The significance of obtained results has also been discussed.

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Estimation of the presence of small dense lipoprotein cholesterol in acute ischemic stroke

Neurology International ◽

10.4081/ni.2015.5973 ◽

2015 ◽

Vol 7 (1) ◽

Cited By ~ 5

Author(s):

Yasuhiro Manabe ◽

Ryuta Morihara ◽

Kosuke Matsuzono ◽

Yumiko Nakano ◽

Yoshiaki Takahashi ◽

...

Keyword(s):

Ischemic Stroke ◽

Acute Ischemic Stroke ◽

Poor Prognosis ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Standard Laboratory ◽

Laboratory Methods ◽

Established Risk Factor ◽

The One ◽

Lipid Parameters

Small dense low-density lipoprotein (sdLDL) is an established risk factor in ischemic heart disease. However, its clinical significance in acute ischemic stroke (AIS) is uncertain. This study evaluates the prognostic value of the presence of sdLDL in patients with AIS by determining whether it contributes to clinical outcome or not. We studied 530 consecutive patients admitted within the first 48 hours after onset of ischemic stroke and 50 corresponding controls. Serum lipid parameters were measured on admission by standard laboratory methods. The percentage of AIS patients with sdLDL was significantly higher than the one of matched controls with sdLDL. Concerning comparisons between AIS patients with or without sdLDL, the percentages of males and patients with histories of smoking, hypertension, and cardiovascular disease were significantly higher in AIS patients with sdLDL. Concerning the grade of severity, modified Rankin Scale (mRS) on discharge was significantly higher in AIS patients with sdLDL. On logistic regression analysis, age (OR=2.29, P3). Our study showed that the presence of sdLDL might be independently associated with a poor prognosis after AIS.

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Low-Bias RNA Sequencing of the HIV-2 Genome from Blood Plasma

Journal of Virology ◽

10.1128/jvi.00677-18 ◽

2018 ◽

Vol 93 (1) ◽

Cited By ~ 3

Author(s):

Katherine L. James ◽

Thushan I. de Silva ◽

Katherine Brown ◽

Hilton Whittle ◽

Stephen Taylor ◽

...

Keyword(s):

Genetic Diversity ◽

Blood Plasma ◽

De Novo ◽

A Priori ◽

Pcr Amplification ◽

Hiv Vaccine ◽

Whole Genome ◽

Plasma Samples ◽

Target Enrichment ◽

Rna Seq

ABSTRACTAccurate determination of the genetic diversity present in the HIV quasispecies is critical for the development of a preventative vaccine: in particular, little is known about viral genetic diversity for the second type of HIV, HIV-2. A better understanding of HIV-2 biology is relevant to the HIV vaccine field because a substantial proportion of infected people experience long-term viral control, and prior HIV-2 infection has been associated with slower HIV-1 disease progression in coinfected subjects. The majority of traditional and next-generation sequencing methods have relied on target amplification prior to sequencing, introducing biases that may obscure the true signals of diversity in the viral population. Additionally, target enrichment through PCR requiresa priorisequence knowledge, which is lacking for HIV-2. Therefore, a target enrichment free method of library preparation would be valuable for the field. We applied an RNA shotgun sequencing (RNA-Seq) method without PCR amplification to cultured viral stocks and patient plasma samples from HIV-2-infected individuals. Libraries generated from total plasma RNA were analyzed with a two-step pipeline: (i)de novogenome assembly, followed by (ii) read remapping. By this approach, whole-genome sequences were generated with a 28× to 67× mean depth of coverage. Assembled reads showed a low level of GC bias, and comparison of the genome diversities at the intrahost level showed low diversity in the accessory genevpxin all patients. Our study demonstrates that RNA-Seq is a feasible full-genomede novosequencing method for blood plasma samples collected from HIV-2-infected individuals.IMPORTANCEAn accurate picture of viral genetic diversity is critical for the development of a globally effective HIV vaccine. However, sequencing strategies are often complicated by target enrichment prior to sequencing, introducing biases that can distort variant frequencies, which are not easily corrected for in downstream analyses. Additionally, detaileda priorisequence knowledge is needed to inform robust primer design when employing PCR amplification, a factor that is often lacking when working with tropical diseases localized in developing countries. Previous work has demonstrated that direct RNA shotgun sequencing (RNA-Seq) can be used to circumvent these issues for hepatitis C virus (HCV) and norovirus. We applied RNA-Seq to total RNA extracted from HIV-2 blood plasma samples, demonstrating the applicability of this technique to HIV-2 and allowing us to generate a dynamic picture of genetic diversity over the whole genome of HIV-2 in the context of low-bias sequencing.

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Plasma and white adipose tissue lipid composition in marmots

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1990.258.5.r1123 ◽

1990 ◽

Vol 258 (5) ◽

pp. R1123-R1131 ◽

Cited By ~ 18

Author(s):

G. L. Florant ◽

L. C. Nuttle ◽

D. E. Mullinex ◽

D. A. Rintoul

Keyword(s):

Adipose Tissue ◽

White Adipose Tissue ◽

Plasma Free Fatty Acid ◽

Laboratory Animals ◽

Plasma Samples ◽

Standard Laboratory ◽

Marmota Flaviventris ◽

Fa Composition ◽

Omega 6 ◽

Standard Laboratory Diet

White adipose tissue biopsies and plasma samples were obtained from hibernating yellow-bellied marmots (Marmota flaviventris) maintained in the laboratory. In addition, biopsies and plasma samples were obtained from normothermic animals in the field and laboratory. Measurement of plasma free fatty acid (FA) levels indicated that winter laboratory animals exhibited increased lipolysis. Additionally, analysis of white adipose tissue triacylglycerol revealed that the FA composition of the storage fat in animals maintained on the standard laboratory diet is remarkably simple and uniform between different adipose depots in the same animal. Three FAs (palmitic, oleic, and linoleic acids) made up greater than 95% of the total. Triene (alpha-linolenate) was found in newly captured animals, but the percentage of this FA decreased rapidly when the animals were maintained on the standard laboratory diet. Throughout the hibernation season (October to April), white adipose tissue-saturated FA percentage decreased, monoene percentage remained constant, and diene percentage increased. Analysis of plasma FA composition suggested that these animals tended to metabolize saturated FAs from stored lipid during hibernation and that dienes were mobilized briefly after the last arousal from hibernation in spring. From these observations, we hypothesize that marmots preferentially metabolize saturated fats during the hibernation period and that essential FAs of the omega 6 series tend to be metabolized more slowly than other FAs. These characteristics suggest that marmots are a valuable animal model in which to study lipid metabolism.

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Monitoring Radiotherapeutic Response in Prostate Cancer Patients Using High Throughput FTIR Spectroscopy of Liquid Biopsies

Cancers ◽

10.3390/cancers11070925 ◽

2019 ◽

Vol 11 (7) ◽

pp. 925 ◽

Cited By ~ 4

Author(s):

Dinesh K.R. Medipally ◽

Thi Nguyet Que Nguyen ◽

Jane Bryant ◽

Valérie Untereiner ◽

Ganesh D. Sockalingum ◽

...

Keyword(s):

Prostate Cancer ◽

Blood Plasma ◽

Ftir Spectroscopy ◽

Cancer Patients ◽

Radiation Toxicity ◽

Plasma Samples ◽

Analysis Model ◽

Liquid Biopsies ◽

Time Points ◽

Wide Range

Radiation therapy (RT) is used to treat approximately 50% of all cancer patients. However, RT causes a wide range of adverse late effects that can affect a patient’s quality of life. There are currently no predictive assays in clinical use to identify patients at risk of normal tissue radiation toxicity. This study aimed to investigate the potential of Fourier transform infrared (FTIR) spectroscopy for monitoring radiotherapeutic response. Blood plasma was acquired from 53 prostate cancer patients at five different time points: prior to treatment, after hormone treatment, at the end of radiotherapy, two months post radiotherapy and eight months post radiotherapy. FTIR spectra were recorded from plasma samples at all time points and the data was analysed using MATLAB software. Discrimination was observed between spectra recorded at baseline versus follow up time points, as well as between spectra from patients showing minimal and severe acute and late toxicity using principal component analysis. A partial least squares discriminant analysis model achieved sensitivity and specificity rates ranging from 80% to 99%. This technology may have potential to monitor radiotherapeutic response in prostate cancer patients using non-invasive blood plasma samples and could lead to individualised patient radiotherapy.

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The Nutritive Value of Commonly Consumed Processed and Unprocessed Vegetables in South-Southern Nigeria

European Journal of Nutrition & Food Safety ◽

10.9734/ejnfs/2019/v11i430174 ◽

2020 ◽

pp. 263-273

Author(s):

Lilian C. Aburime ◽

Onot O. Ekpe ◽

Gift O. Okpuruwu ◽

Chidera T. Nweke

Keyword(s):

Social Science ◽

Dietary Fibre ◽

Nutrient Intake ◽

Nutritive Value ◽

Chemical Analyses ◽

Standard Laboratory ◽

Laboratory Methods ◽

Significant Difference ◽

Southern Nigeria ◽

Mineral Compositions

Aim: The nutritive value, proximate, vitamin and mineral compositions of seven commonly consumed vegetables in South-Southern Nigeria were determined. Methodology: Pods and leaves of vegetables were divided into two parts. One part was analyzed raw and the remaining part was cooked by boiling. The raw vegetable leaves and okro pods were separately washed and dried using a food dehydrator (40ºC) for 24hrs. After drying, it was milled into fine flour using an electric blender. The second part of the vegetables were processed by boiling using variable time, after boiling, they were separately drained and dried using a food dehydrator at a temperature of about 50ºC for 48 hrs. Chemical analyses were carried out using standard laboratory methods. Means and standard deviations were calculated using the statistical package for social science. The least significant difference (LSD) was accepted at P = .05 significance. Results: For the raw vegetables the results were as follows: Moisture 71.2 – 91.2%, ash 0.9- 2.9%, dietary fibre 9.2 – 13.1%, fat 0.3 -1.4%, protein 2.1 – 8.2%, available Carbohydrate 1.0 – 8.7%, phosphorus 8 -52 mg/100 g, potassium 263- 1152 mg/100 g, sodium 3-23 mg/100 g, calcium 183-815 mg/100 g, magnesium 67-217 mg/100 g, iron 0.26-1.27 mg/100 g, zinc 0.26-1.10 mg, folate 3-13 mcg/100 g. The results of the boiled vegetables were: moisture 80.9– 93.8%, protein 1.7 – 3.2%, fat 0.2 – 0.5%, ash 0.5– 1.1%, dietary fibre 3.9 – 9.3%, available carbohydrate 3.8 – 9.9%, phosphorus 7 – 35 mg/100 g, Potassium 0.33 – 300 mg/100 g, sodium 0.3 – 20 mg/100 g, magnesium 45 – 132 mg/100 g, calcium 82 – 200 mg/100 g, iron 0.21 -0.60 mg/100 g, zinc 0.13 – 0.30mg/100g. Folate 3 – 6 mcg/100 g. The range of the percentage contributions of the vegetables that are consumed raw to the recommended nutrient intake or recommended dietary allowance of adults are as follows: protein 5-18%, fat 1-3%, carbohydrates 1-6%, dietary fibre 3-52%, iron 2-14%, zinc 6-36%, calcium 26 -116%, phosphorus 8-31%, sodium 0.3-1%, Potassium 8-32%, Magnesium 26-99%, Folate 3-4%. Conclusion: Boiling significantly reduced most nutrient studied. Bitter leaves are more nutrient dense than other studied vegetables. Knowing the food compositions of these vegetables will promote their use.

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CREATININE AND CREATINE KINASE RATIO IN BLOOD OF DIFFERENT BODY TYPES - A NEW APPROACH.

STADIUM - Hungarian Journal of Sport Sciences ◽

10.36439/shjs/2021/2/10561 ◽

2021 ◽

Vol 4 (2) ◽

Author(s):

Anupam Bandyopadhyay ◽

Pritha Chatterjee

Keyword(s):

Creatine Kinase ◽

Creatinine Level ◽

Creatine Kinase Level ◽

Fat Content ◽

Standard Laboratory ◽

Laboratory Methods ◽

New Approach ◽

Modifiable Factors ◽

Post Hoc ◽

One Way Anova

Purpose: The inconsistencies and variations of creatine kinase level due to modifiable and non-modifiable factors were the basis of this study. The aim was to find out the relationships between creatinine and creatine kinase in the blood of somatotypes. Methods: The 122 males, aged 10 to 20 years, were classified according to their somatotypes. Somatotypes were measured by the ISAK method. By standard laboratory methods, creatinine and creatine kinase estimate. The IBM SPSS version 24 is used for calculation. One way ANOVA followed by post hoc tests was performed to compare the variables among the three groups (p<0.05). Results: Creatinine level in the blood insignificantly deferred among the three somatotypes. The significant differences (p<0.05) were found in creatine kinase level in the blood and creatinine/creatine kinase ratio among the three dominant Somatotypes. Creatine Kinase was significantly higher in Ectomorphs (212 U/L) than Endomorphs. Ectomorphs and mesomorphs have crossed normal creatine kinase levels (35 -175 U/L). The creatinine/creatine kinase ratio was found highest in endomorphs and lowest in the ectomorphs and significantly differed in three Somatotypes. Conclusion: Creatinine production remains the same, indicating production of Creatinine is independent of specific body types. A significant higher Creatine Kinase level in Ectomorphs over Endomorphs showed fat content was not associated with it. Significant differences in Creatinine / Creatine Kinase ratio among Somatotypes suggested its relevance between cellular and morphological relationships and might uses as biomarkers.

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