841 results on '"BLOOD sampling"'
Search Results
2. Molecular prevalence, associated risk factors and phylogenetic evaluation of Theileria lestoquardi in the blood samples of small ruminants.
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Ashraf, Sehrish, Hikal, Wafaa M., Almahallawi, Ruoa, Muqaddas, Hira, and Iqbal, Furhan
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BLOOD sampling , *RUMINANTS , *THEILERIA , *ANIMAL herds , *CELL surface antigens , *RUMEN fermentation , *SHEEP breeding , *DAIRY farm management - Abstract
Raising small ruminants is the main source of income for farmers in Pakistan especially in rural areas of Dera Ghazi Khan in Punjab. Despite having large sheep population, the prevalence of intra-erythrocytic protozoa, Theileria (T.) lestoquardi, has never been reported from this area. This study was conducted to fill this knowledge gap and 333 blood samples of apparently healthy small ruminants (168 sheep and 165 goats) along with their epidemiological data were collected from Dera Ghazi Khan district during August till November 2022. The polymerase chain reaction (PCR) analysis amplified a 785 base pair amplicon specific for the Merozoite surface antigen (ms 1–2) gene of T. lestoquardi in 2 out of the 168 (3.3%) sheep blood samples, while no goat blood sample out of 165 was found to be infected with T. lestoquardi. DNA sequencing confirmed the presence of Theileria lestoquardi in both samples and phylogenetic analysis revealed that these amplicon resembled the partial ms 1–2 gene sequences detected in small ruminants from Pakistan, India Iran and Egypt. All the studied epidemiological factors (age, sex, breed, size of herd, dogs with herd, composition of herd, size of herd and Tick burden on sheep) were not found associated with the prevalence of T. lestoquardi. In conclusion, this study reports a low prevalence of T. lestoquardi infection in the Dera Ghazi Khan District of Punjab, Pakistan. The data generated from this work will help pave the way for the prophylactic detection and control of ovine and caprine theileriosis in the region. [ABSTRACT FROM AUTHOR]
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- 2024
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3. Drugs in blood and urine samples from victims of suspected exposure to drink spiking: A prospective observational study from Oslo, Norway.
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Dalaker, Vivian M., Furuhaugen, Håvard, Brekke, Mette, Bjørnaas, Mari Asphjell, Krpo, Maja, Øiestad, Elisabeth Leere, and Vallersnes, Odd Martin
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URINALYSIS , *LIQUID chromatography-mass spectrometry , *BLOOD sampling , *BEVERAGES , *DRUG abuse , *LONGITUDINAL method - Abstract
Objective: People regularly contact emergency medicine services concerned that they have been exposed to drink spiking, i.e., exposure to drugs without their knowledge or permission. We identified drugs in blood and urine samples from patients suspecting exposure to drink spiking, with special consideration for drugs not reported taken by the patient (unreported drugs). Methods: From September 2018 to May 2019, we collected blood and urine samples from patients 16 years or older presenting at an emergency clinic in Oslo, Norway, within 48 hours of suspected exposure to drink spiking. We also collected information on ethanol ingestion and drugs taken. Blood samples were analyzed for 20 classical recreational drugs using ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) and an automated enzymatic method for ethanol. Urine samples were analyzed using immunoassay methods and a specific gas chromatography mass spectrometry (GCMS) method for gammahydroxybutyrate (GHB). Results: From 100 included patients (median age 24 years, 62 females), we collected 100 blood samples and 72 urine samples. Median time since exposure was 5 hours. Unreported drugs were found in 15 patients. Unreported drugs in the blood samples were clonazepam in 3, methylenedioxymethamphetamine (MDMA) in 3, amphetamine in 2, tetrahydrocannabinol (THC) in 2, tramadol in 1, cocaine in 1, and methamphetamine in 1. Unreported drugs in the urine samples were cocaine in 5, amphetamine in 4, ecstasy in 3, and cannabis in 2. Ethanol was found in 69 patients, all reporting ethanol ingestion. Median blood ethanol concentration was higher in patients with no unreported drugs detected, 1.00‰ (interquartile range (IQR) 0–1.52) vs. 0‰ (IQR 0–0.46) (p<0.001). GHB was not detected. Conclusion: Unreported drugs, possibly used for drink spiking, were found in 15% of patients. Blood ethanol concentration was higher when no unreported drugs were found. GHB was not detected in any patient. [ABSTRACT FROM AUTHOR]
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- 2024
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4. Feasibility, acceptability, and safety of a novel device for self-collecting capillary blood samples in clinical trials in the context of the pandemic and beyond.
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Dasari, Harika, Smyrnova, Anna, Leng, Jing, and Ducharme, Francine M.
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PAIN perception , *BLOOD volume , *BLOOD sampling , *CLINICAL trials , *CAPILLARIES , *PAIN management - Abstract
Background: Home blood self-collection devices can enable remote monitoring, but their implementation requires validation. Our objectives were to explore (i) the impact of sampling sites and topical analgesia on capillary blood volume and pain perception and (ii) the safety, acceptability, and failure of capillary self-collection among adults and children using the Tasso-SST device. Methods: We conducted a two-phase study. The investigational phase consisted of two on-site cross-sectional studies in healthy adult participants (≥ 12 years) and children (1–17 years) with their accompanying parent. Adults received 4 capillary samplings, where puncture sites and topical analgesia were randomized in a factorial design, and a venipuncture; children (and one parent) had one capillary sampling. The two co-primary outcomes were blood volume and pain. The implementation phase was conducted in two multicentre trials in participants choosing remote visits; blood volume, collection failure, adverse events, and satisfaction were documented. Results: In the investigational phase, 90 participants and 9 children with 7 parents were enrolled; 15 adults and 2 preschoolers participated in the implementation phase. In the adult investigational study, the device collected a median (25%, 75%) of 450 (250, 550) μl of blood with no significant difference between the puncture site, topical analgesia, and its interaction. Using topical analgesia reduced pain perception by 0.61 (95% CI: 0.97, 0.24; P <0.01) points on the 11-point scale; the pain reduction varied by puncture site, with the lower back showing the most significant decrease. Overall, combining all studies and phases, the median volume collected was 425 (250, 500) μl, and the device failure rate was 4.4%; minor adverse effects were reported in 8.9% of the participants, all were willing to use the device again. Conclusion: Capillary blood self-collection, yielding slightly less than 500 μl, proves to be a safe and relatively painless method for adults and children, with high satisfaction and low failure rates. The puncture site and topical analgesia do not affect blood volume, but topical analgesia on the lower back could reduce pain. [ABSTRACT FROM AUTHOR]
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- 2024
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5. Age estimation of captive Asian elephants (Elephas maximus) based on DNA methylation: An exploratory analysis using methylation-sensitive high-resolution melting (MS-HRM).
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Arai, Kana, Qi, Huiyuan, and Inoue-Murayama, Miho
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ASIATIC elephant , *DNA methylation , *AGE , *MELTING , *BLOOD sampling - Abstract
Age is an important parameter for bettering the understanding of biodemographic trends—development, survival, reproduction and environmental effects—critical for conservation. However, current age estimation methods are challenging to apply to many species, and no standardised technique has been adopted yet. This study examined the potential use of methylation-sensitive high-resolution melting (MS-HRM), a labour-, time-, and cost-effective method to estimate chronological age from DNA methylation in Asian elephants (Elephas maximus). The objective of this study was to investigate the accuracy and validation of MS-HRM use for age determination in long-lived species, such as Asian elephants. The average lifespan of Asian elephants is between 50–70 years but some have been known to survive for more than 80 years. DNA was extracted from 53 blood samples of captive Asian elephants across 11 zoos in Japan, with known ages ranging from a few months to 65 years. Methylation rates of two candidate age-related epigenetic genes, RALYL and TET2, were significantly correlated with chronological age. Finally, we established a linear, unisex age estimation model with a mean absolute error (MAE) of 7.36 years. This exploratory study suggests an avenue to further explore MS-HRM as an alternative method to estimate the chronological age of Asian elephants. [ABSTRACT FROM AUTHOR]
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- 2023
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6. Development of a sensitive real-time quaking-induced conversion (RT-QuIC) assay for application in prion-infected blood.
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Thomas, Charlotte M., Salamat, M. Khalid F., de Wolf, Christopher, McCutcheon, Sandra, Blanco, A. Richard Alejo, Manson, Jean C., Hunter, Nora, and Houston, E. Fiona
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CREUTZFELDT-Jakob disease , *BOVINE spongiform encephalopathy , *PRION diseases , *HUMAN-to-human transmission , *FERRIC oxide , *BLOOD sampling - Abstract
Efforts to prevent human-to-human transmission of variant Creutzfeldt-Jakob disease (vCJD) by contaminated blood would be aided by the development of a sensitive diagnostic test that could be routinely used to screen blood donations. As blood samples from vCJD patients are extremely rare, here we describe the optimisation of real-time quaking-induced conversion (RT-QuIC) for detection of PrPSc (misfolded prion protein, a marker of prion infection) in blood samples from an established large animal model of vCJD, sheep experimentally infected with bovine spongiform encephalopathy (BSE). Comparative endpoint titration experiments with RT-QuIC, miniaturized bead protein misfolding cyclic amplification (mb-PMCA) and intracerebral inoculation of a transgenic mouse line expressing sheep PrP (tgOvARQ), demonstrated highly sensitive detection of PrPSc by RT-QuIC in a reference sheep brain homogenate. Upon addition of a capture step with iron oxide beads, the RT-QuIC assay was able to detect PrPSc in whole blood samples from BSE-infected sheep up to two years before disease onset. Both RT-QuIC and mb-PMCA also demonstrated sensitive detection of PrPSc in a reference vCJD-infected human brain homogenate, suggesting that either assay may be suitable for application to human blood samples. Our results support the further development and evaluation of RT-QuIC as a diagnostic or screening test for vCJD. [ABSTRACT FROM AUTHOR]
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- 2023
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7. New erythrocyte parameters derived from the Coulter principle relate with red blood cell properties—A pilot study in diabetes mellitus.
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Bourguignon, Chloé, Ansel, Clémentine, Gineys, Jean-Philippe, Schuldiner, Sophie, Isèbe, Damien, Geitner, Michael, Taraconat, Pierre, and Gris, Jean-Christophe
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BLOOD cell count , *DIABETES , *PILOT projects , *ERYTHROCYTES , *BLOOD sampling , *PEOPLE with diabetes - Abstract
In routine hematological instruments, blood cells are counted and sized by monitoring the impedance signals induced during their passage through a Coulter orifice. However, only signals associated with centered paths in the aperture are considered for analysis, while the rejected measurements, caused by near-wall trajectories, can provide additional information on red blood cells (RBC), as recent publications suggest. To assess usefulness of two new parameters in describing alterations in RBC properties, we performed a pilot study to compare blood samples from patients with diabetes mellitus (DM), frequent pathological condition associated with impairment in RBC deformability, versus controls. A total of 345 blood samples were analyzed: 225 in the DM group and 120 in the control group. A diagram of R and P , the two new parameters derived from the analysis of impedancemetry pulses, was used to compare distribution of RBC subpopulations between groups. To discriminate RBC from DM and control individuals, based on our multiparametric analysis, we built a score from variables derived from R/P matrix which showed good performances: area under the receiving operating characteristic curve 0.948 (0.920–0.970), p<0.0001; best discriminating value: negative predictive value 94.7%, positive predictive value was 78.4%. These results seem promising to approach RBC alterations in routine laboratory practice. The related potential clinically relevant outcomes remain to be investigated. [ABSTRACT FROM AUTHOR]
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- 2023
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8. Nicotine flux as a powerful tool for regulating nicotine delivery from e-cigarettes: Protocol of two complimentary randomized crossover clinical trials.
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El-Hellani, Ahmad, Hanna, Elyana, Sharma, Mehak, Blohowiak, Reagan, Joseph, Phillip, Eid, Tore, Nadim, Haleh, El-Hage, Rachel, Salman, Rola, Karaoghlanian, Nareg, Adeniji, Ayomipo, Salam, Sally, Talih, Farid, Elbejjani, Martine, Breland, Alison, Eissenberg, Thomas, Shihadeh, Alan, Baldassarri, Stephen R., and Talih, Soha
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CIGARETTES , *CLINICAL trials , *NICOTINE , *ELECTRONIC cigarettes , *BLOOD sampling , *DRUG withdrawal symptoms - Abstract
Introduction: Electronic cigarette (EC) use has increased rapidly in the last decade, especially among youth. Regulating nicotine delivery from ECs could help curb youth uptake and leverage EC use in harm reduction yet is complicated by varying device and liquid variables that affect nicotine delivery. Nicotine flux, the nicotine emission rate, is a parameter that incorporates these variables and focuses on the performance rather than the design of an EC. Nicotine flux therefore could be a powerful regulatory tool if it is shown empirically to predict nicotine delivery and subjective effects related to dependence. Methods and analysis: This project consists of two complementary clinical trials. In Trial I, we will examine the relationship between nicotine flux and the rate and dose of nicotine delivery from ECs, hence, impacting abuse liability. It will also examine the extent to which this relationship is mediated by nicotine form (i.e., freebase versus protonated). At Yale School of Medicine (YSM), study participants will puff EC devices under conditions that differ by flux and form, while arterial blood is sampled in high time resolution. In Trial II, we will assess the relationship between nicotine flux, form, and subjective effects. At the American University of Beirut (AUB), participants will use EC devices with varying nicotine fluxes and forms, while dependency measures, such as the urge to use ECs, nicotine craving, and withdrawal symptoms, will be assessed. We will also monitor puffing intensity and real-time exposure to toxicants. Ethics and dissemination: The protocol of Trial I and Trial II was approved by YSM and AUB IRBs, respectively. We will disseminate study results through peer-reviewed publications and conference presentations. Trial registration: NCT05706701 for Trial I and NCT05430334 for Trial II. [ABSTRACT FROM AUTHOR]
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- 2023
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9. Pre-analytical variables influence zinc measurement in blood samples.
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Killilea, David W. and Schultz, Kathleen
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INDUCTIVELY coupled plasma atomic emission spectrometry , *BLOOD sampling , *ZINC , *SAMPLING (Process) , *BLOOD collection , *CARBOXYHEMOGLOBIN - Abstract
Zinc deficiency continues to be a major concern for global public health. The zinc status of a target population is typically estimated by measuring circulating zinc levels, but the sampling procedures are not standardized and thus may result in analytical discrepancies. To examine this, we designed a study that controlled most of the technical parameters in order to focus on five pre-analytical variables reported to influence the measurement of zinc in blood samples, including (1) blood draw site (capillary or venous), (2) blood sample matrix (plasma or serum), (3) blood collection tube manufacturer (Becton, Dickinson and Company or Sarstedt AG & Co), (4) blood processing time (0, 4, or 24 hours), and (5) blood holding temperatures (4°C, 20°C, or 37°C). A diverse cohort of 60 healthy adults were recruited to provide sequential capillary and venous blood samples, which were carefully processed under a single chain of custody and measured for zinc content using inductively coupled plasma optical emission spectrometry. When comparing blood draw sites, the mean zinc content of capillary samples was 0.054 mg/L (8%; p<0.0001) higher than venous blood from the same donors. When comparing blood sample matrices, the mean zinc content of serum samples was 0.029 mg/L (5%; p<0.0001) higher than plasma samples from the same donors. When comparing blood collection tube manufacturer, the mean zinc content from venous blood samples did not differ between venders, but the mean zinc content from BD capillary plasma was 0.036 mg/L (6%; p<0.0001) higher than Sarstedt capillary plasma from the same donors. When comparing processing times, the mean zinc content of plasma and serum samples was 5–12% higher (p<0.0001) in samples processed 4–24 hour after collection. When comparing holding temperatures, the mean zinc content of plasma and serum samples was 0.5–7% higher (p = 0.0007 or p = 0.0061, respectively) in samples temporarily held at 20°C or 37°C after collection. Thus even with the same donors and blood draws, significant differences in zinc content were observed with different draw sites, tube types, and processing procedures, demonstrating that key pre-analytic variables can have an impact on zinc measurement, and subsequent classification of zinc status. Minimizing these pre-analytical variables is important for generating best practice guidelines for assessment of zinc status. [ABSTRACT FROM AUTHOR]
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- 2023
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10. Commercial hatchery processing may affect susceptibility to stress in laying hens.
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Van Poucke, Enya, Suchánková, Hedvika, and Jensen, Per
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HENS , *HATCHERY fishes , *LIFE change events , *CHICKS , *BODY temperature , *BLOOD sampling , *PSYCHOLOGICAL stress - Abstract
Directly upon hatching, laying hen chicks are exposed to multiple stressful events during large-scale hatchery processing, which may affect their later coping abilities. Commercial hatchery chicks (HC) were compared to chicks that were incubated and hatched simultaneously under calm conditions (CC). After being raised under similar, non-stressful conditions for 36 days, all chicks were exposed to a series of stressors: transportation and introduction into a novel environment followed by a regrouping event in order to characterize long-lasting consequences of hatchery treatment. Tonic immobility, corticosterone levels, and peripheral body temperature were used to assess reactions to the stress events. Tonic immobility was not affected by treatment but was significantly reduced in CC after transport. Corticosterone levels did not differ between treatments when assessed two days before and two days after regrouping. Comb temperature was significantly higher in HC following regrouping, indicating stress-induced hyperthermia. Furthermore, comb temperature dropped more following blood sampling in HC than in CC, indicating a stronger autonomic response to acute stress. In conclusion, the results suggest possible long-term negative effects of commercial hatchery processing, compared to hatching under silent and less stressful conditions, on the coping ability of laying hens to later stressful experiences. [ABSTRACT FROM AUTHOR]
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- 2023
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11. A sensitive tissue factor activity assay determined by an optimized thrombin generation method.
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Kristensen, Søren Risom and Nybo, Jette
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THROMBIN , *BLOOD volume , *THROMBOEMBOLISM , *EXTRACELLULAR vesicles , *BLOOD sampling - Abstract
Background: Tissue factor (TF) is the principal activator of the coagulation system, but an increased concentration in the blood in cancer and inflammatory diseases has been suggested to play a role increasing the risk of venous thromboembolism. However, measurement of the TF concentration is difficult, and quantitation of activity is the most valid estimation. The objective of this study was to establish a sensitive method to measure TF activity based on thrombin generation. Methods: The assay is based on thrombin generation (TG) measured on the Calibrated Automated Thrombogram (CAT). Various low concentrations of TF were prepared from reagents containing 1 pM TF and 4 μM phospholipid (PPL), and no TF and 4 μM PPL, and a calibration curve was produced from Lagtime vs TF concentration. TF in blood samples was measured after isolation and resuspension of extracellular vesicles (EVs) in a standard plasma from which EVs had been removed. The same standard plasma was used for the calibrators. Results: Contact activation of the coagulation system was avoided using CTI plasma samples in Monovette tubes. EVs contain procoagulant phospholipids but addition of PPL only reduced lagtime slightly at very low concentrations of TF resulting in overestimation to a lesser extent at 10 fM but no interference at 30 fM or higher. Addition of EVs to the TG analysis induced a small unspecific TF-independent activity (i.e., an activity not inhibited by antibodies against TF) which also may result in a smaller error in estimation of TF activity at very low levels but the effect was negligible at higher concentrations. It was possible to measure TF activity in healthy controls which was found to be 1–6 fM (EVs were concentrated, i.e. solubilized in a lower volume than the original volume plasma). Coefficient of variation (CV) was below 20% at the low level, and below 10% at a level around 100 fM TF. However, the step with isolation of EVs have a higher inherent CV. Conclusion: A sensitive and rather precise one-stage TG-based method to measure TF activity has been established. [ABSTRACT FROM AUTHOR]
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- 2023
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12. Simultaneous molecular detection of Anaplasma marginale and Theileria annulata in cattle blood samples collected from Pakistan-Afghanistan boarder region.
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Jamil, Sania, Chiou, Chien-Chun, Muqaddas, Hira, Ullah, Hayat, Asif, Muhammad, Rao, Sana, Hussain, Hafsa, Fatima, Qandeel, Nasreen, Nasreen, Niaz, Sadaf, Dzul-Rosado, Karla, Khan, Adil, Iqbal, Furhan, and Chen, Chien-Chin
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THEILERIA , *ANAPLASMA marginale , *BLOOD sampling , *CATTLE , *LIVESTOCK productivity , *TICK control - Abstract
Theileria annulata (T. annulata) and Anaplasma marginale (A. marginale) are among the most extensively reported tick borne pathogens and are associated with huge economic losses worldwide. A total of 298 cattle blood samples were screened to report the presence of these two pathogens. The samples were collected from apparently healthy cattle (Achai, n = 155, Jersy, n = 88 and crossbred, n = 55) in Bajaur district of Khyber Pakhtunkhwa (KPK) during June and July of 2022. A total of 31 out of 298 cattle (10.4%) were found infected with T. annulata as PCR amplified a 156 base pair fragment from Tams-1 gene of T. annulata from their blood. While 16/298 animals (5.4%) were found infected with A. marginale as they amplified a 382 base pair fragment specific for msp5 gene of this bacterium. Three animals (1%) were found co infected. Cattle susceptibility to T. annulata infection was significantly higher than A. marginale infection (P < 0.001). Phylogenetic analysis revealed that Pakistani isolates of both detected pathogen clustered together and were closely related isolates from worldwide countries. Prevalence of T. annulata varied significantly among the sampling sites (P = 0.05) while no such association was observed for A. marginale among the tested cattle. Epidemiological data analysis revealed that none of the studied risk factors was found associated either with the prevalence of T. annulata or A. marginale (P > 0.05) among enrolled cattle. In conclusion, our study has revealed a relatively higher prevalence of T. annulata than A. marginale in cattle from the Bajaur district in KPK. This information is important for improving the productivity of the livestock sector, which is one of the main sources of income in the country. It is recommended that this data be taken into account for the development and implementation of effective tick control programs, as well as for the improvement of livestock management practices to prevent and manage TBDs in Pakistan. [ABSTRACT FROM AUTHOR]
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- 2023
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13. Feasibility of self-organized blood sample collection in adults for study purposes in a primary care setting.
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Schröder, Dominik, Müller, Frank, Heesen, Gloria, Hummers, Eva, Dopfer-Jablonka, Alexandra, Vahldiek, Kai, Klawonn, Frank, Steffens, Sandra, Mikuteit, Marie, Niewolik, Jacqueline, and Heinemann, Stephanie
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BLOOD collection , *BLOOD sampling , *COVID-19 , *MEDICAL personnel , *PRIMARY care , *CORD blood - Abstract
Background/aims: The COVID-19 pandemic situation poses new challenges for research. Ethical issues might arise if especially vulnerable individuals for severe COVID-19 course expose themselves because of participation in studies to a higher risk of infection for study purposes. How is the feasibility and acceptance of self-organized blood sample collections to measure anti-SARS-CoV-2 Spike IgG antibodies in persons with a high risk for a severe COVID-19 disease progression? Methods: Persons with a high risk for a severe COVID-19 disease progression (immunocompromised, oncology patients or over 80 years old) were recruited between January and September 2021 to send in blood samples (at least 500 μl) 1 month and 6 months after second COVID-19 vaccination. Participants were given the choice of drawing capillary or venous blood themselves or having blood drawn by health professionals belonging to either the study's own research team or the personnel found in local practices or clinics. Participants were surveyed via a telephone interview in December 2021 and January 2022 about their choice of blood sampling methods and influence of blood collection choice upon study participation. Results: Data from 360 participants was collected via telephone follow-up. First blood samples were collected by the participants themselves (35.8%), local practices or clinics (31.9%) and the research team (22.5%). Second blood samples were mostly collected in local practices or clinics (35.6%) followed by participants themselves (25.9%) and the research team (11.5%). Blood samples were not collected in 2.5% and 19.1% of persons during first and second blood draw, respectively. Only 2% of blood samples did not reach the laboratory or were not analyzable. About one-fourth (26%) of participants stated that they would not have participated in the study if it would have been required to travel to the university hospital to give their blood sample. Conclusions: Participants were able to self-organize blood collection, making use of several different blood sample methods. Nearly all blood samples were analyzable when self-collected and sent in by post. One-fourth of the participants would not have participated in the study if required to give their blood sample in the study location. Trial registration: German Clinical Trial Registry, DRKS00021152. [ABSTRACT FROM AUTHOR]
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- 2023
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14. Remote Assessment in healthcare—Technologies, methods, benefits, and challenges.
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Bardram, Jakob Eyvind
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TELEMEDICINE , *TECHNOLOGY assessment , *PRENATAL diagnosis , *BLOOD sampling , *MEDICAL care , *ORAL health - Abstract
The PLOS ONE Collection on "Remote Assessment" brings together a series of studies on how remote assessment methods and technologies can be used in health and behavioral sciences. At the time of writing (October 2022), this collection has accepted and published 10 papers, which address remote assessment in a wide range of health topics including mental health, cognitive assessment, blood sampling and diagnosis, dental health, COVID-19 infections, and prenatal diagnosis. The papers also cover a wide range of methodological approaches, technology platforms, and ways to utilize remote assessment. As such, this collection provides a broad view into the benefits and challenges of remote assessment, and provides a lot of detailed knowledge on how to make it work in practice This paper provides an overview of the included studies, and presents and discusses the different benefits as well as challenges associated with remote assessment. [ABSTRACT FROM AUTHOR]
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- 2023
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15. Low doses of diarrhoeagenic E. coli induce enhanced monocyte and mDC responses and prevent development of symptoms after homologous rechallenge.
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Porbahaie, Mojtaba, van den Belt, Maartje, Ulfman, Laurien, Ruijschop, Rianne M. A. J., Lucas–van de Bos, Elly, Hartog, Anita, Lenz, Stefanie, van Alen-Boerrigter, Ingrid J., Teodorowicz, Malgorzata, Savelkoul, Huub F. J., Calame, Wim, van Hoffen, Els, van Neerven, R. J. Joost, and Kardinaal, Alwine
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ESCHERICHIA coli , *CALPROTECTIN , *SYMPTOMS , *BLOOD sampling , *DEFECATION , *IMMUNE response - Abstract
The experimental challenge with attenuated enterotoxigenic E. coli strain E1392/75-2A prevents diarrhea upon a secondary challenge with the same bacteria. A dose-response pilot study was performed to investigate which immunological factors are associated with this protection. Healthy subjects were inoculated with increasing E. coli doses of 1E6-1E10 CFU, and three weeks later, all participants were rechallenged with the highest dose (1E10 CFU). Gastrointestinal discomfort symptoms were recorded, and stool and blood samples were analyzed. After the primary challenge, stool frequency, diarrhea symptom scores, and E. coli-specific serum IgG (IgG-CFA/II) titer increased in a dose-dependent manner. Fecal calprotectin and serum IgG-CFA/II response after primary challenge were delayed in the lower dose groups. Even though stool frequency after the secondary challenge was inversely related to the primary inoculation dose, all E. coli doses protected against clinical symptoms upon rechallenge. Ex vivo stimulation of PBMCs with E. coli just before the second challenge resulted in increased numbers of IL-6+/TNF-α+ monocytes and mDCs than before the primary challenge, without dose-dependency. These data demonstrate that primary E. coli infection with as few as 1E6 CFU protects against a high-dose secondary challenge with a homologous attenuated strain. Increased serum IgG-CFA/II levels and E. coli-induced mDC and monocyte responses after primary challenge suggest that protection against secondary E. coli challenges is associated with adaptive as well as innate immune responses. [ABSTRACT FROM AUTHOR]
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- 2023
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16. A novel RP-HPLC method for quantification of cholinesterase activity in human blood: An application for assessing organophosphate and carbamate insecticide exposure.
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Sinha, Sukesh Narayan, Ungarala, Ramakrishna, Kumar, Dileshwar, Sangaraju, Rajendra, and Kumar, Sarita
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CHOLINESTERASE reactivators , *HIGH performance liquid chromatography , *ACETYLCHOLINESTERASE , *INSECTICIDES , *SPRAYING , *BLOOD sampling , *PESTICIDES , *ACETATES - Abstract
Several methods have been reported to estimate Acetylcholinesterase (AChE) enzyme activity in blood samples. The Ellman assay is the most important among all but with several shortcomings, and there is a need to develop a method which is accurate, sensitive and quick for analyzing AChE. Therefore, we have developed an assay utilizing RP-HPLC with UV detection for the determination of AChE activity. This method measured the conversion of 1-naphthol acetate to 1-naphthol to estimate AChE activity in blood samples. Performance was judged on the basis of reproducibility, sensitivity, accuracy, and the ability to screen enzyme activity within 20minutes. A series of experiments were performed, varying the concentration of blood and substrate, with optimal sensitivity using 50 μM substrate and 10μL blood. The validation parameters such as linearity (R2 of ≥ 0.9842 for 1-naphthol and ≥ 0.9897 for 1-naphthol acetate), precision (94.21–96.41%), accuracy (85.2%–99.6% and 82.6%–99.3% for 1-naphthol and 1-naphthol acetate respectively), and robustness were validated according to International Conference on Harmonization (ICH) guidelines. Blood samples were collected from healthy people, farmers exposed to spraying of pesticides, and suicidal patients who ingested pesticides and were hospitalized and were analyzed by the developed method. The AChE level was approximately 21 units/mL compared to 24units/mL in controls, whereas suicidal patients showed the least AChE levels of 1 unit/mL. The employment of this method is recommended for estimating AChE level on various matrices. [ABSTRACT FROM AUTHOR]
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- 2022
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17. Risk factors and outcome due to extended-spectrum β-lactamase-producing uropathogenic Escherichia coli in community-onset bloodstream infections: A ten-year cohort study in Sweden.
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Holmbom, Martin, Möller, Vidar, Kristinsdottir, Loa, Nilsson, Maud, Rashid, Mamun-Ur, Fredrikson, Mats, Berglund, Björn, and Östholm Balkhed, Åse
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ESCHERICHIA coli , *COHORT analysis , *ANTIBIOTICS , *INFECTION , *BLOOD sampling - Abstract
Objective: To study clinical outcome and risk factors associated with extended-spectrum β-lactamase (ESBL)-producing uropathogenic Escherichia coli (UPEC) in community-onset bloodstream infections (CO-BSI). Methods: This was a population-based cohort study including patients with pheno- and genotype-matched ESBL-producing E. coli and non-ESBL- E. coli in urine and blood samples collected in 2009–2018 in southeast Sweden. Seventy-seven episodes of ESBL-UPEC satisfying the inclusion criteria were matched 1:1 with 77 non-ESBL-UPEC for age, gender, and year of culture. Results: The most common ST-type and ESBL gene was ST131 (55%), and blaCTX-M-15 (47%), respectively. Risk factors for ESBL-UPEC were: previous genitourinary invasive procedure (RR 4.66; p = 0.005) or history of ESBL-producing E. coli (RR 12.14; p = 0.024). There was significant difference between ESBL-UPEC and non-ESBL-UPEC regarding time to microbiologically appropriate antibiotic therapy (27:15 h vs. 02:14 h; p = <0.001) and hospital days (9 vs. 5; p = <0.001), but no difference in 30-day mortality (3% vs. 3%; p = >0.999) or sepsis within 36 hours (51% vs. 62%; p = 0.623) was observed. Conclusion: The predominant risk factors for ESBL-UPEC were history of ESBL-Ec infection and history of genitourinary invasive procedure. The overall mortality was low and the delay in appropriate antibiotic therapy did not increase the risk for 30-day mortality or risk for sepsis within 36 hours among patients infected with ESBL UPEC. However, these results must be regarded with some degree of caution due to the small sample size. [ABSTRACT FROM AUTHOR]
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- 2022
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18. Absorption rate of subcutaneously infused fluid in ill multimorbid older patients.
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Danielsen, Mathias Brix, Jødal, Lars, Riis, Johannes, Karmisholt, Jesper Scott, Valdórsson, Óskar, Jørgensen, Martin Gronbech, and Andersen, Stig
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OLDER patients , *FLUIDS , *ABSORPTION , *THYROID gland , *BLOOD sampling , *SAMPLE size (Statistics) - Abstract
Background: Subcutaneous (SC) hydration is a valuable method for treating dehydration in the very old patients. Data are absent on the absorption rate, and the availability of SC infused fluid in the circulation in this group of patients where SC hydration is particularly relevant. Methods: We performed an explorative study on ill very old (range 78–84 years old) geriatric patients with comorbidities who received an SC infusion of 235 ml isotonic saline containing a technetium-99m pertechnetate tracer. The activity over the infusion site was measured using a gamma detector to assess the absorption rate from the SC space. The activity was measured initially every 5 minutes, with intervals extended gradually to 15 minutes. Activity in blood samples and the thyroid gland was measured to determine the rate of availability in the circulation. Results: Six patients were included. The mean age was 81 years (SD 2.1), the number of comorbidities was 4.6 (SD 1.3), and the Tilburg frailty indicator was 3.8 (SD 2.4). When the infusion was completed after 60 minutes, 53% (95% CI 50–56%) of the infused fluid was absorbed from the SC space, with 88% (95% CI 86–90%) absorbed one hour later. The absorption rate from the SC space right after the completion of the infusion was 127 ml/h (95% CI 90–164 ml/h). The appearance of the fluid into the blood and the thyroid gland verified the transfer from SC to circulation. Conclusion: This first explorative study of absorption of SC infused fluid in the very old found an acceptable amount of fluid absorbed from the SC space into the circulation one hour after infusion had ended. Results are uniform but should be interpreted cautiously due to the low sample size. Trial registration: ClinicalTrials.gov Identifier: NCT04536324. [ABSTRACT FROM AUTHOR]
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- 2022
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19. Analytical and clinical characterization of an optimized dual monoclonal sandwich ELISA for the quantification of thymidine kinase 1 (TK1) protein in human blood samples.
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Jagarlamudi, K. K., L., Swinkels, M., Zupan, J., Osredkar, P., Venge, and S., Eriksson
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BLOOD proteins , *THYMIDINE , *MONOCLONAL antibodies , *BLOOD sampling , *HEMATOLOGIC malignancies , *DNA synthesis - Abstract
Thymidine Kinase 1 (TK1) plays an important role in DNA precursor synthesis and serum TK1 activity has been used as a biomarker for prognosis and therapy monitoring of different malignancies. AroCell has developed a dual monoclonal antibody ELISA for determination of TK1 protein in clinical samples. The purpose of the study is to validate the ELISA analytically in relation to the gold standard, [3H]-deoxythymidine (dThd) phosphorylation assay for TK1 activity using sera from patients with different malignancies. The colorimetric TK 210 ELISA was validated analytically by assessment of precision, linearity, interfering substances, and stability. For the clinical validation, serum samples from patients with hematological malignancies (n = 100), breast cancer (n = 56), prostate cancer (n = 70) and blood donors (n = 159) were analyzed using TK 210 ELISA and TK1 activity by [3H]-deoxythymidine (dThd) phosphorylation assay. The sandwich TK 210 ELISA was highly specific for TK1 protein having a detection limit of 0.12 ng/mL, with a functional sensitivity of 0.25 ng/mL. Within-run CVs ranged from 5.5% to 10% and between-run CVs ranged from 5% to 15%. The ratio of observed to expected dilutional parallelism of 5 serum samples was in the range of 80–120%. Samples exhibited stability through four freeze/thaw cycles and 5 days at 4°C. Further, the ROC curve analysis showed that TK 210 ELISA and [3H]-dThd phosphorylation assay had similar sensitivity (62% vs 59%) in hematological malignancies. However, in the case of breast and prostate cancer sera, TK 210 ELISA had higher sensitivity (59% and 44%) compared to [3H]-dThd phosphorylation assay (47% and 25%) at a specificity of 98%. These data demonstrate that the dual monoclonal antibody based AroCell TK 210 ELISA is a robust, accurate and precise tool for measuring TK1 protein in different malignancies that can improve the clinical applications of TK1 as a biomarker in cancer management. [ABSTRACT FROM AUTHOR]
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- 2022
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20. Influence of drone carriage material on maintenance of storage temperature and quality of blood samples during transportation in an equatorial climate.
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Zailani, Mohamed Afiq Hidayat, Raja Sabudin, Raja Zahratul Azma, Ismail, Aniza, Abd Rahman, Rahana, Mohd Saiboon, Ismail, Sabri, Shahnaz Irwani, Seong, Chan Kok, Mail, Jamaludin, Md Jamal, Shamsuriani, Beng, Gan Kok, and Mahdy, Zaleha Abdullah
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BLOOD sampling , *FOAM , *BLOOD testing , *DRONE aircraft delivery , *TEMPERATURE , *STORAGE - Abstract
The disruptive potentials of drones are rapidly growing including for the delivery of blood samples in healthcare. Maintenance of the quality of blood samples is important to ascertain that the drone is a safe mode of transportation, particularly during emergencies and in critical cases. The influence of the drone carriage material on blood samples transportation was investigated in this study. Two phases of drone simulation flights were conducted in Cyberjaya, Malaysia. In Phase 1, the effect of drone carriage material on the internal storage temperature during blood samples transportation was determined. Three types of carriage materials were compared: aluminium, expanded polystyrene (EPS) foam, and polypropylene (PP) plastic. In Phase 2, the quality of drone-transported blood samples was assessed, using the best material from Phase 1 as the drone carriage material. Biochemical and hematological analyses of 60 blood samples were conducted using five parameters. In Phase 1, EPS foam was found to be the best material to maintain a stable and favorable internal storage temperature at mean kinetic temperature ±SD of 4.70 ±1.14°C. Much higher and unfavorable mean kinetic temperatures were recorded for aluminium (11.46 ±0.35°C) and plastic (14.17 ±0.05°C). In Phase 2, laboratory tests show that the quality of blood samples was well maintained, and the mean biochemical and hematological parameters of drone-transported blood samples showed no significant alteration compared to ground controls. Drone carriage material is an important determinant of the quality of blood samples transported by drone, particularly in hot equatorial climates as in Malaysia. The blood storage temperature was best maintained using EPS foam, as evidenced by the favorable average temperature and preservation of hematological and biochemical parameters of the blood samples. [ABSTRACT FROM AUTHOR]
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- 2022
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21. Apelin and its ratio to lipid factors are associated with cardiovascular diseases: A systematic review and meta-analysis.
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Akbari, Hamed, Hosseini-Bensenjan, Mahnaz, Salahi, Sarvenaz, Moazzen, Fatemeh, Aria, Hamid, Manafi, Alireza, Hosseini, Saeed, Niknam, Maryam, and Asadikaram, Gholamreza
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APELIN , *CARDIOVASCULAR diseases , *PEOPLE with diabetes , *CONFIDENCE intervals , *BLOOD sampling - Abstract
Background: The present systematic review and meta-analysis aimed to ascertain if the circulating levels of apelin, as an important regulator of the cardiovascular homeostasis, differ in patients with cardiovascular diseases (CVDs) and controls. Methods: A comprehensive search was performed in electronic databases including PubMed, Scopus, EMBASE, and Web of Science to identify the studies addressing apelin in CVD up to April 5, 2021. Due to the presence of different units to measure the circulating levels of apelin across the included studies, they expressed the standardized mean difference (SMD) and their 95% confidence interval (CI) as summary effect size. A random-effects model comprising DerSimonian and Laird method was used to pool SMDs. Results: Twenty-four articles (30 studies) comprised of 1793 cases and 1416 controls were included. Pooled results obtained through random-effects model indicated that apelin concentrations in the cases' blood samples were significantly lower than those of the control groups (SMD = -0.72, 95% CI: -1.25, -0.18, P = 0.009; I2 = 97.3%, P<0.001). New combined biomarkers showed a significant decrease in SMD of apelin/high-density lipoprotein cholesterol (apelin/HDL-C) ratio [-5.17; 95% CI, -8.72, -1.63, P = 0.000; I2 = 99.0%], apelin/low-density lipoprotein cholesterol (apelin/LDL-C) ratio [-4.31; 95% CI, -6.08, -2.55, P = 0.000; I2 = 98.0%] and apelin/total cholesterol (apelin/TC) ratio [-17.30; 95% CI, -22.85, -11.76, P = 0.000; I2 = 99.1%]. However, no significant differences were found in the SMD of apelin/triacylglycerol (apelin/TG) ratio in cases with CVDs compared to the control group [-2.96; 95% CI, -7.41, 1.49, P = 0.000; I2 = 99.2%]. Conclusion: The association of apelin with CVDs is different based on the region and disease subtypes. These findings account for the possible usefulness of apelin as an additional biomarker in the diagnosis of CVD in diabetic patients and in the diagnosis of patients with CAD. Moreover, apelin/HDL-c, apelin/LDL-c, and apelin/TC ratios could be offered as diagnostic markers for CVD. [ABSTRACT FROM AUTHOR]
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- 2022
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22. A low-cost, open-source centrifuge adaptor for separating large volume clinical blood samples.
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Haque, Md Ehtashamul, Marriott, Linda, Naeem, Noman, Henry, Taygan, Conde, Alvaro J., and Kersaudy-Kerhoas, Maïwenn
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BLOOD volume , *BLOOD sampling , *CENTRIFUGES , *HELLP syndrome , *BLOOD plasma , *THREE-dimensional printing - Abstract
Blood plasma separation is a prerequisite in numerous biomedical assays involving low abundance plasma-borne biomarkers and thus is the fundamental step before many bioanalytical steps. High-capacity refrigerated centrifuges, which have the advantage of handling large volumes of blood samples, are widely utilized, but they are bulky, non-transportable, and prohibitively expensive for low-resource settings, with prices starting at $1,500. On the other hand, there are low-cost commercial and open-source micro-centrifuges available, but they are incapable of handling typical clinical amounts of blood samples (2-10mL). There is currently no low-cost CE marked centrifuge that can process large volumes of clinical blood samples on the market. As a solution, we customised the rotor of a commercially available low-cost micro-centrifuge (~$125) using 3D printing to enable centrifugation of large clinical blood samples in resource poor-settings. Our custom adaptor ($15) can hold two 9 mL S-Monovette tubes and achieve the same separation performance (yield, cell count, hemolysis, albumin levels) as the control benchtop refrigerated centrifuge, and even outperformed the control in platelet separation by at least four times. This low-cost open-source centrifugation system capable of processing clinical blood tubes could be valuable to low-resource settings where centrifugation is required immediately after blood withdrawal for further testing. [ABSTRACT FROM AUTHOR]
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- 2022
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23. Genetically regulated gene expression and proteins revealed discordant effects.
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Pott, Janne, Garcia, Tarcyane, Hauck, Stefanie M., Petrera, Agnese, Wirkner, Kerstin, Loeffler, Markus, Kirsten, Holger, Peters, Annette, and Scholz, Markus
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PROTEIN expression , *GENE expression , *GENETIC regulation , *GENETIC correlations , *BLOOD sampling - Abstract
Background: Although gene-expression (GE) and protein levels are typically strongly genetically regulated, their correlation is known to be low. Here we investigate this phenomenon by focusing on the genetic background of this correlation in order to understand the similarities and differences in the genetic regulation of these omics layers. Methods and results: We performed locus-wide association studies of 92 protein levels measured in whole blood for 2,014 samples of European ancestry and found that 66 are genetically regulated. Three female- and one male-specific effects were detected. We estimated the genetically regulated GE for all significant genes in 49 GTEx v8 tissues. A total of 7 proteins showed negative correlations with their respective GE across multiple tissues. Finally, we tested for causal links of GE on protein expression via Mendelian Randomization, and confirmed a negative causal effect of GE on protein level for five of these genes in a total of 63 gene-tissue pairs: BLMH, CASP3, CXCL16, IL6R, and SFTPD. For IL6R, we replicated the negative causal effect on coronary-artery disease (CAD), while its GE was positively linked to CAD. Conclusion: While total GE and protein levels are only weakly correlated, we found high correlations between their genetically regulated components across multiple tissues. Of note, strong negative causal effects of tissue-specific GE on five protein levels were detected. Causal network analyses revealed that GE effects on CAD risks was in general mediated by protein levels. [ABSTRACT FROM AUTHOR]
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- 2022
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24. Serological and molecular detection of Babesia caballi and Theileria equi in Mexico: A prospective study.
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Salinas-Estrella, Elizabeth, Ueti, Massaro W., Lobanov, Vladislav A., Castillo-Payró, Evelio, Lizcano-Mata, Amelia, Badilla, César, Martínez-Ibáñez, Francisco, and Mosqueda, Juan
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BABESIA , *THEILERIA , *HORSE diseases , *BABESIOSIS , *LONGITUDINAL method , *VIRAL antibodies , *BLOOD sampling ,TROPICAL climate - Abstract
Equine piroplasmosis is a disease of horses, mules and donkeys, caused by the hemoprotozoans Babesia caballi and Theileria equi and transmitted by ticks of tropical and subtropical regions. Because the clinical signs are not specific, the diagnosis of equine piroplasmosis is difficult. In Mexico, where the environmental factors are conducive to the persistence of these pathogens, there is a lack of molecular studies to evaluate the occurrence of both parasites in horses. In the present study, matching serum and whole blood samples were obtained from 269 horses residing in 24 locations with tropical or subtropical climate and the presence of ticks. Testing of serum samples by ELISA demonstrated 55.7% seroprevalence of B. caballi and 68.4% prevalence of antibodies to T. equi. Blood samples analyzed with nPCR test were 7.8% positive to B. caballi and 78.8% positive to T. equi, while a duplex qPCR showed 15.24% positive samples to B. caballi and 59.11% to T. equi. From these results, 27 samples were sequenced for T. equi and 13 for B. caballi, confirming the presence of both horse parasites that cause equine piroplasmosis and suggesting that they are widespread in Mexico. This is the first study confirming the presence of B. caballi and T. equi in Mexico using both serological and molecular diagnostic methods. This study shows a high incidence of exposure to the etiological agents of equine piroplasmosis in horses in the studied areas. [ABSTRACT FROM AUTHOR]
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- 2022
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25. Evaluation of urine sample for diagnosis of visceral leishmaniasis using rK-39 immunochromatographic test in Northwest Ethiopia.
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Eyayu, Tahir, Yasin, Melashu, Workineh, Lemma, Tiruneh, Tegenaw, Andualem, Henok, Sema, Meslo, Damtie, Shewaneh, Abebaw, Aynework, Getie, Birhanu, Andargie, Desalegn, Achaw, Barnabas, and Taklual, Wubet
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VISCERAL leishmaniasis , *LEISHMANIASIS , *URINE , *CONTINGENCY tables , *BLOOD sampling , *DIAGNOSIS - Abstract
Background: Visceral leishmaniasis is the most severe form of leishmaniasis which ranks second in mortality and fourth in morbidity. Parasitological diagnostic techniques with splenic aspirate remain the gold standard. However, sample collection is risky, painful, and difficult. Alternatively, serological techniques provide good diagnostic accuracy using serum sample that is difficult for applying on small children and in the field. So, finding alternative non-invasive and self-collected samples like urine is very important. Thus, the study aimed to evaluate the diagnostic performance of the rK-39 strip test using urine for diagnosis of visceral leishmaniasis. Methods: A multicenter institutional-based cross-sectional study was conducted from November 2019 to March 2021 at Northwest Ethiopia. Sociodemographic information was collected using a structured questionnaire. Blood sample and midstream urine sample were collected for rK-39 test. Data were entered into Epi-data version 4.2 and analyzed using SPSS version 24.0. Diagnostic performance parameters of urine-based rK-39 rapid test, i.e. sensitivity, specificity, positive and negative predictive values, positive and negative likelihood ratios (LR+/−), and diagnostic accuracy were determined on contingency table by using serum-based rK-39 test result as a reference. An agreement between urine and serum-based rK-39 test was statistically determined by kappa value. Result: In total, 300 subjects, age ranged between 7 and 60 years, were included in the study. The overall sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of urine-based rK-39 test were found to be 98.0% (95% CI: 93.0% - 99.8%), 95.5% (95% CI: 91.6% - 97.9%), 91.6% (95% CI: 85.2%– 95.4%), 98.9 (95% CI: 96.0%– 99.7%), and 96.33% (95% CI: 93.53–98.16%), respectively. Additionally, there was a strong agreement between the results obtained on rK-39 ICT using urine and serum samples (kappa = 0.92; P < 0.001). Conclusion: Urine-based rK-39 ICT had an excellent high sensitivity, specificity and strong agreement with serum-based rK-39 ICT results. This indicates that urine sample would be a promising noninvasive and easy to collect sample for diagnosis of VL in field and rural settings. [ABSTRACT FROM AUTHOR]
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- 2022
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26. Sex differences in serum levels of 5α-androstane-3β, 17β-diol, and androstenediol in the young adults: A liquid chromatography–tandem mass spectrometry study.
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Tanabe, Haruka, Mutai, Hitoshi, Sasayama, Daimei, Sasamoto, Hidehiko, Miyashiro, Yoshimichi, Sugiyama, Nobuhiro, and Washizuka, Shinsuke
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MENSTRUAL cycle , *LIQUID chromatography-mass spectrometry , *SEX factors in disease , *HAMILTON Depression Inventory , *YOUNG adults , *BLOOD sampling - Abstract
Animal experiments have consistently shown that estrogen receptor β (ERβ)-selective ligands have antidepressant and anxiolytic effects. In humans, endogenous ligands for ERβ include 5α-androstane-3β, 17β-diol (3βAdiol) and androstenediol (Δ5-diol). We determined, for the first time, the exact serum levels of 3βAdiol and Δ5-diol in young healthy volunteers using liquid chromatography–tandem mass spectrometry (LC–MS/MS). We investigated the effect of the menstrual cycle on the levels of these steroids in women; then, we performed a gender comparison. Blood samples were collected from 48 subjects: 23 women (mean age = 28.4±7.8 years) and 25 men (mean age = 31.4±7.8 years). We collected the blood samples of women at three time-points in the menstrual cycle: the early follicular phase, ovulatory or mid-cycle phase, and mid-luteal phase. A total of 92 blood samples were analyzed using LC–MS/MS. The levels of two well-studied steroids, namely dehydroepiandrosterone (DHEA) and 17β-estradiol (E2), were simultaneously measured. Depression rating scale (Hamilton Rating Scale for Depression, Beck Depression Inventory-II and Quick Inventory of Depressive Symptomatology) scores were also recorded at the time of blood sampling. Significant differences in the levels of 3βAdiol and E2 and in the depression rating scale scores were observed over the duration of the menstrual cycle of the women. The levels of 3βAdiol and Δ5-diol were significantly lower in women than in men. E2 levels were higher in women than in men, and DHEA levels did not differ significantly between men and women. Further, women had higher scores than men on the Hamilton Rating Scale for Depression. Sex differences in depressive symptoms can be explained by 3βAdiol and Δ5-diol levels, and the effect of the menstrual cycle on mood can be explained by 3βAdiol and E2 levels, not by Δ5-diol level. [ABSTRACT FROM AUTHOR]
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- 2021
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27. Analyses of blood donor samples from eight provinces in Lao PDR suggest considerable variation concerning HBV exposure and carriage.
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Nouanthong, Phonethipsavanh, Hefele, Lisa, Keokhamphue, Jerapha, Sorrasin, Vonhphet, Khounvisith, Vilaysone, Souksakhone, Chanthala, Jutavijittum, Prapan, Muller, Claude P., Black, Antony P., and Hübschen, Judith M.
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HEPATITIS associated antigen , *BLOOD testing , *DISEASE prevalence , *BLOOD donors , *BLOOD group antigens , *BLOOD sampling - Abstract
Introduction: Hepatitis B is endemic in Lao PDR and about 9% of the adult population is chronically infected. In this study, we investigated regional, occupational, age and sex-related differences in hepatitis B epidemiology in Lao blood donors. Methods: 5017 voluntary blood donors from 8 different provinces were tested for hepatitis B markers by ELISA. Predictors for the prevalence of hepatitis B surface antigen (HBsAg) and antibodies against the core antigen (anti-HBc) were assessed by bivariate and multivariable analyses. Results: In total, 41% of the participants were positive for anti-HBc; the HBsAg prevalence was estimated at 6.9% among all participants (9.2% among first-time donors and 3.9% among repeat donors). Among first-time donors, HBsAg positivity was associated independently with being male (p<0.001), being from the North (p<0.001) and being soldier (p<0.001). Participants were more likely to be anti-HBc positive when they were male (p<0.001), from the Northern provinces (p<0.001) and older than 20 years (p<0.01). Conclusion: In conclusion, our study confirmed an overall high HBsAg and anti-HBc prevalence in Lao PDR, albeit with considerable regional variation. The identification of a sizeable number of HBsAg positives among repeat donors warrants a thorough investigation of current blood screening, record keeping, donor identification and counselling practises. [ABSTRACT FROM AUTHOR]
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- 2021
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28. Development of at-home sample collection logistics for large-scale seroprevalence studies.
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Aatresh, Aishani V., Cummings, Kate, Gerstein, Hilary, Knight, Christopher S., Limberopolous, Andreas, Stasi, Megan A., Bedugnis, Alice, Somberg, Kenneth A., França, Camila T., and Mina, Michael J.
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DEMOGRAPHIC characteristics , *COVID-19 , *PATIENT selection , *COLLECTIONS , *BLOOD sampling , *PANDEMICS - Abstract
Background: Serological studies rely on the recruitment of representative cohorts; however, such efforts are specially complicated by the conditions surrounding the COVID19 pandemic. Methods: We aimed to design and implement a fully remote methodology for conducting safe serological surveys that also allow for the engagement of representative study populations. Results: This design was well-received and effective. 2,066 participants ≥18 years old were enrolled, reflecting the ethnic and racial composition of Massachusetts. >70% of them reported being satisfied/extremely satisfied with the online enrollment and at-home self-collection of blood samples. While 18.6% reported some discomfort experienced with the collection process, 72.2% stated that they would be willing to test weekly if enrolled in a long-term study. Conclusions: High engagement and positive feedback from participants, as well as the quality of self-collected specimens, point to the usefulness of this fully remote, self-collection-based study design for future safer and efficient population-level serological surveys. [ABSTRACT FROM AUTHOR]
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- 2021
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29. Validation study of Boil & Spin Malachite Green Loop Mediated Isothermal Amplification (B&S MG-LAMP) versus microscopy for malaria detection in the Peruvian Amazon.
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Barazorda, Keare A., Salas, Carola J., Braga, Greys, Ricopa, Leonila, Ampuero, Julia S., Siles, Crystyan, Sanchez, Juan F., Montano, Silvia, Lizewski, Stephen E., Joya, Christie A., Bishop, Danett K., and Valdivia, Hugo O.
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GENE amplification , *MALACHITE green , *MALARIA , *MICROSCOPY , *TURNAROUND time , *PARASITEMIA , *BLOOD sampling - Abstract
Malaria elimination efforts in Peru have dramatically reduced the incidence of cases in the Amazon Basin. To achieve the elimination, the detection of asymptomatic and submicroscopic carriers becomes a priority. Therefore, efforts should focus on tests sensitive enough to detect low-density parasitemia, deployable to resource-limited areas and affordable for large screening purposes. In this study, we assessed the performance of the Malachite–Green LAMP (MG-LAMP) using heat-treated DNA extraction (Boil & Spin; B&S MG-LAMP) on 283 whole blood samples collected from 9 different sites in Loreto, Peru and compared its performance to expert and field microscopy. A real-time PCR assay was used to quantify the parasite density. In addition, we explored a modified version of the B&S MG-LAMP for detection of submicroscopic infection in 500 samples and compared the turnaround time and cost of the MG-LAMP with microscopy. Compared to expert microscopy, the genus B&S MG-LAMP had a sensitivity of 99.4% (95%CI: 96.9%– 100%) and specificity of 97.1% (95%CI: 91.9%– 99.4%). The P. vivax specific B&S MG-LAMP had a sensitivity of 99.4% (96.6%– 100%) and specificity of 99.2% (95.5%– 100%) and the P. falciparum assay had a sensitivity of 100% (95%CI: 78.2%– 100%) and specificity of 99.3% (95%CI: 97.3%– 99.8%). The modified genus B&S MG-LAMP assay detected eight submicroscopic malaria cases (1.6%) which the species-specific assays did not identify. The turnaround time of B&S MG-LAMP was faster than expert microscopy with as many as 60 samples being processed per day by field technicians with limited training and utilizing a simple heat-block. The modified B&S MG-LAMP offers a simple and sensitive molecular test of choice for the detection of submicroscopic infections that can be used for mass screening in resources limited facilities in endemic settings nearing elimination and where a deployable test is required. [ABSTRACT FROM AUTHOR]
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- 2021
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30. Raman spectroscopy on blood serum samples of patients with end-stage liver disease.
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Staritzbichler, René, Hunold, Pascal, Estrela-Lopis, Irina, Hildebrand, Peter Werner, Isermann, Berend, and Kaiser, Thorsten
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SERUM , *LIVER diseases , *RAMAN spectroscopy , *BLOOD sampling , *SUBSET selection , *MEDICAL laboratories - Abstract
Raman spectroscopy has shown to be a promising method for the examination of biomedical samples. However, until now, its efficacy has not been established in clinical diagnostics. In this study, Raman spectroscopy's potential application in medical laboratories is evaluated for a large variety (38) of biomarkers. Given 234 serum samples from a cohort of patients with different stages of liver disease, we performed Raman spectroscopy at 780nm excitation wavelength. The Raman spectra were analyzed in combination with the results of routine diagnostics using specifically developed complex mathematical algorithms, including fluorescence filtering, frequency subset selection and several overfitting circumventing strategies, such as independent validation. With the results of this cohort, which were validated in 328 independent samples, a significant proof-of-concept study was completed. This study highlights the need to prevent overfitting and to use independent data for validation. The results reveal that Raman spectroscopy has high potential for use in medical laboratory diagnostics to simultaneously quantify multiple biomarkers. [ABSTRACT FROM AUTHOR]
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- 2021
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31. Point-of-care microvolume cytometer measures platelet counts with high accuracy from capillary blood.
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Dickerson, William M., Yu, Rebecca, Westergren, Helena U., Paraskos, Jonathan, Schatz, Philipp, Tigerstrom, Anna, Ekman, Anna, Sánchez, José, Cheng, Jamie, Li, Lillian, and Chan, Eugene Y.
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PLATELET count , *POINT-of-care testing , *ADULTS , *CAPILLARIES , *BLOOD sampling - Abstract
Background: Point-of-care (PoC) testing of platelet count (PLT) provides real-time data for rapid decision making. The goal of this study is to evaluate the accuracy and precision of platelet counting using a new microvolume (8 μL), absolute counting, 1.5 kg cytometry-based blood analyzer, the rHEALTH ONE (rHEALTH) in comparison with the International Society of Laboratory Hematology (ISLH) platelet method, which uses a cytometer and an impedance analyzer. Methods: Inclusion eligibility were healthy adults (M/F) ages 18–80 for donation of fingerprick and venous blood samples. Samples were from a random N = 31 volunteers from a single U.S. site. Samples were serially diluted to test thrombocytopenic ranges. Interfering substances and conditions were tested, including RBC fragments, platelet fragments, cholesterol, triglycerides, lipids, anti-platelet antibodies, and temperature. Results: The concordance between the rHEALTH and ISLH methods had a slope = 1.030 and R2 = 0.9684. The rHEALTH method showed a correlation between capillary and venous blood samples (slope = 0.9514 and R2 = 0.9684). Certain interferents changed platelet recovery: RBC fragments and anti-platelet antibodies with the ISLH method; platelet fragments and anti-platelet antibodies on the rHEALTH; and RBC fragments, platelets fragments, triglycerides and LDL on the clinical impedance analyzer. The rHEALTH's precision ranged from 3.1–8.0%, and the ISLH from 1.0–10.5%. Conclusions: The rHEALTH method provides similar results with the reference method and good correlation between adult capillary and venous blood samples. This demonstrates the ability of the rHEALTH to provide point-of-care assessment of normal and thrombocytopenic platelet counts from fingerprick blood with high precision and limited interferences. [ABSTRACT FROM AUTHOR]
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- 2021
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32. Variation of B cell subsets with age in healthy Malawians.
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Mandala, Wilson L. and Longwe, Herbert
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CELLULAR aging , *LYMPHOCYTE subsets , *B cells , *AGE groups , *ETHNICITY , *BLOOD sampling , *POPULATION aging - Abstract
Although a number of previous studies have shown that different lymphocyte subsets, including B cells, vary with age, how different B cell subsets vary with age in Malawian population has not been shown before. We recruited Malawian participants of different ages and analyzed their venous blood samples for different B cell subsets. We found that both percentage and absolute counts of B cells varied with age peaking in the 7 to 12 months age group. Proportion of naïve B cells was highest in neonates and decreased with age whereas the percentage of memory B cells was lowest in neonates and increased with age. When we zeroed in on the age band within which the proportion of B cells was highest, both classical and activated memory B cells increased with age and the naïve followed the opposite trend. These results provide additional knowledge in our understanding of the dynamics of B cell subsets in individuals of a specific ethnicity as they age. [ABSTRACT FROM AUTHOR]
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- 2021
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33. Does varying the ingestion period of sodium citrate influence blood alkalosis and gastrointestinal symptoms?
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Urwin, Charles S., Snow, Rodney J., Orellana, Liliana, Condo, Dominique, Wadley, Glenn D., and Carr, Amelia J.
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SYMPTOMS , *CITRATES , *SODIUM , *GELATIN , *BLOOD sampling - Abstract
Objectives: To compare blood alkalosis, gastrointestinal symptoms and indicators of strong ion difference after ingestion of 500 mg.kg-1 BM sodium citrate over four different periods. Methods: Sixteen healthy and active participants ingested 500 mg.kg-1 BM sodium citrate in gelatine capsules over a 15, 30, 45 or 60 min period using a randomized cross-over experimental design. Gastrointestinal symptoms questionnaires and venous blood samples were collected before ingestion, immediately post-ingestion, and every 30 min for 480 min post-ingestion. Blood samples were analysed for blood pH, [HCO3-], [Na+], [Cl-] and plasma [citrate]. Linear mixed models were used to estimate the effect of the ingestion protocols. Results: For all treatments, blood [HCO3-] was significantly elevated above baseline for the entire 480 min post-ingestion period, and peak occurred 180 min post-ingestion. Blood [HCO3-] and pH were significantly elevated above baseline and not significantly below the peak between 150–270 min post-ingestion. Furthermore, blood pH and [HCO3-] were significantly lower for the 60 min ingestion period when compared to the other treatments. Gastrointestinal symptoms were minor for all treatments; the mean total session symptoms ratings (all times summed together) were between 9.8 and 11.6 from a maximum possible rating of 720. Conclusion: Based on the findings of this investigation, sodium citrate should be ingested over a period of less than 60 min (15, 30 or 45 min), and completed 150–270 min before exercise. [ABSTRACT FROM AUTHOR]
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- 2021
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34. Fit-for-purpose quantitative liquid biopsy based droplet digital PCR assay development for detection of programmed cell death ligand-1 (PD-L1) RNA expression in PAXgene blood samples.
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O'Rourke, Dennis, Wang, Danyi, Sanchez-Garcia, Jorge F., Cusano, Maria Perella, Miller, Waldemar, Cai, Ti, Scheuenpflug, Juergen, and Feng, Zheng
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CIRCULATING tumor DNA , *APOPTOSIS , *PROGRAMMED death-ligand 1 , *BLOOD sampling , *INTERFERON gamma , *RNA - Abstract
Development of a clinically applicable liquid biopsy-based test for PD-L1 mRNA expression would be beneficial in providing complementary evidence to current immunohistochemistry assays. Hence, we report the development of a fit-for-purpose assay for detection of blood PD-L1 mRNA expression using droplet digital polymerase chain reaction (ddPCR). TaqMan® assays were selected based on coverage of the PD-L1 gene and were tested for linearity and efficiency using real-time quantitative PCR. Four reference genes were analyzed in positive control cell line (A549 treated with interferon gamma, [IFN γ]) genomic DNA. The PD-L1 primer/probe sets were also evaluated in ddPCR for limit of blank, limit of detection, and precision. Finally, thirty-five healthy volunteer samples were evaluated to establish a baseline level of PD-L1 expression. In ddPCR, the limit of blank was determined to be 0 copies and the limit of detection was determined to be less than or equal to 19 copies of PD-L1. The average intra-run coefficient of variation in the ddPCR assay was 7.44% and average inter-run CV was 7.70%. Treatment of A549 cells with IFN γ resulted in a 6.7-fold increase in PD-L1 expression (21,580 copies in untreated cDNA versus 145,000 copies in treated cDNA). Analysis of healthy human samples yielded a median value of 1659 PD-L1 copies/μL with a range of 768–7510 copies/μL. The assay was transferred to an external service provider and results from our in-house experiments and those conducted externally shows a correlation of 0.994. In conclusion, a fit-for-purpose liquid biopsy-based, purely quantitative ddPCR assay for the detection of PD-L1 mRNA expression was developed and validated using PAXgene RNA blood samples. Linearity, reproducibility, limit of blank and limit of detection were measured and deemed suitable for clinical application. This ultra-sensitive liquid biopsy ddPCR assay has promising clinical potential in screening, longitudinal monitoring and disease progression detection. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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35. Automatic real-time analysis and interpretation of arterial blood gas sample for Point-of-care testing: Clinical validation.
- Author
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Rodríguez-Villar, Sancho, Poza-Hernández, Paloma, Freigang, Sascha, Zubizarreta-Ormazabal, Idoia, Paz-Martín, Daniel, Holl, Etienne, Pérez-Pardo, Osvaldo Ceferino, Tovar-Doncel, María Sherezade, Wissa, Sonja Maria, Cimadevilla-Calvo, Bonifacio, Tejón-Pérez, Guillermo, Moreno-Fernández, Ismael, Escario-Méndez, Alejandro, Arévalo-Serrano, Juan, Valentín, Antonio, Do-Vale, Bruno Manuel, Fletcher, Helen Marie, and Lorenzo- Fernández, Jesús Medardo
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BLOOD gases , *ACID-base imbalances , *BLOOD sampling , *POINT-of-care testing , *MEDICAL personnel - Abstract
Background: Point-of-care arterial blood gas (ABG) is a blood measurement test and a useful diagnostic tool that assists with treatment and therefore improves clinical outcomes. However, numerically reported test results make rapid interpretation difficult or open to interpretation. The arterial blood gas algorithm (ABG-a) is a new digital diagnostics solution that can provide clinicians with real-time interpretation of preliminary data on safety features, oxygenation, acid-base disturbances and renal profile. The main aim of this study was to clinically validate the algorithm against senior experienced clinicians, for acid-base interpretation, in a clinical context. Methods: We conducted a prospective international multicentre observational cross-sectional study. 346 sample sets and 64 inpatients eligible for ABG met strict sampling criteria. Agreement was evaluated using Cohen's kappa index, diagnostic accuracy was evaluated with sensitivity, specificity, efficiency or global accuracy and positive predictive values (PPV) and negative predictive values (NPV) for the prevalence in the study population. Results: The concordance rates between the interpretations of the clinicians and the ABG-a for acid-base disorders were an observed global agreement of 84,3% with a Cohen's kappa coefficient 0.81; 95% CI 0.77 to 0.86; p < 0.001. For detecting accuracy normal acid-base status the algorithm has a sensitivity of 90.0% (95% CI 79.9 to 95.3), a specificity 97.2% (95% CI 94.5 to 98.6) and a global accuracy of 95.9% (95% CI 93.3 to 97.6). For the four simple acid-base disorders, respiratory alkalosis: sensitivity of 91.2 (77.0 to 97.0), a specificity 100.0 (98.8 to 100.0) and global accuracy of 99.1 (97.5 to 99.7); respiratory acidosis: sensitivity of 61.1 (38.6 to 79.7), a specificity of 100.0 (98.8 to 100.0) and global accuracy of 98.0 (95.9 to 99.0); metabolic acidosis: sensitivity of 75.8 (59.0 to 87.2), a specificity of 99.7 (98.2 to 99.9) and a global accuracy of 97.4 (95.1 to 98.6); metabolic alkalosis sensitivity of 72.2 (56.0 to 84.2), a specificity of 95.5 (92.5 to 97.3) and a global accuracy of 93.0 (88.8 to 95.3); the four complex acid-base disorders, respiratory and metabolic alkalosis, respiratory and metabolic acidosis, respiratory alkalosis and metabolic acidosis, respiratory acidosis and metabolic alkalosis, the sensitivity, specificity and global accuracy was also high. For normal acid-base status the algorithm has PPV 87.1 (95% CI 76.6 to 93.3) %, and NPV 97.9 (95% CI 95.4 to 99.0) for a prevalence of 17.4 (95% CI 13.8 to 21.8). For the four-simple acid-base disorders and the four complex acid-base disorders the PPV and NPV were also statistically significant. Conclusions: The ABG-a showed very high agreement and diagnostic accuracy with experienced senior clinicians in the acid-base disorders in a clinical context. The method also provides refinement and deep complex analysis at the point-of-care that a clinician could have at the bedside on a day-to-day basis. The ABG-a method could also have the potential to reduce human errors by checking for imminent life-threatening situations, analysing the internal consistency of the results, the oxygenation and renal status of the patient. [ABSTRACT FROM AUTHOR]
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- 2021
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36. Novel three-dimensional biochip pulmonary sarcoidosis model.
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Calcagno, Tess M., Zhang, Chongxu, Tian, Runxia, Ebrahimi, Babak, and Mirsaeidi, Mehdi
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SARCOIDOSIS , *CYTOKINES , *EOSINOPHILIC granuloma , *BLOOD sampling , *MICROFLUIDICS - Abstract
Sarcoidosis is a multi-system disorder of granulomatous inflammation which most commonly affects the lungs. Its etiology and pathogenesis are not well defined in part due to the lack of reliable modeling. Here, we present the development of an in vitro three-dimensional lung-on-chip biochip designed to mimic granuloma formation. A lung on chip fluidic macrodevice was developed and added to our previously developed a lung-on-membrane model (LOMM). Granulomas were cultured from blood samples of patients with sarcoidosis and then inserted in the air-lung-interface of the microchip to create a three-dimensional biochip pulmonary sarcoidosis model (3D BSGM). Cytokines were measured after 48 hours. ELISA testing was performed to measure cytokine response difference between LOMM with 3D BSGM. There were statistically significant differences in IL-1ß (P = 0.001953), IL-6 (P = 0.001953), GM-CSF (P = 0.001953), and INF-γ expressions (P = 0.09375) between two groups. The current model represents the first 3D biochip sarcoidosis model created by adding a microfluidics system to a dual-chambered lung on membrane model and introducing developed sarcoid-granuloma to its air-lung-interface. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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37. Enumerating regulatory T cells in cryopreserved umbilical cord blood samples using FOXP3 methylation specific quantitative PCR.
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Duggleby, Richard C., Tsang, Hoi Pat, Strange, Kathryn, McWhinnie, Alasdair, Lamikanra, Abigail A., Roberts, David J., Hernandez, Diana, Madrigal, J. Alejandro, and Danby, Robert D.
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SUPPRESSOR cells , *CRYOPRESERVATION of cells , *BLOOD sampling , *CORD blood , *CELL death , *METHYLATION , *FLOW cytometry - Abstract
Background: Allogeneic haematopoietic cell transplantation (HCT) is a curative therapy for severe haematological disorders. However, it carries significant risk of morbidity and mortality. To improve patient outcomes, better graft selection strategies are needed, incorporating HLA matching with clinically important graft characteristics. Studies have shown that the cellular content of HCT grafts, specifically higher ratios of T regulatory (Tregs)/T cells, are important factors influencing outcomes when using adult peripheral blood mobilised grafts. So far, no equivalent study exists in umbilical cord blood (CB) transplantation due to the limitations of cryopreserved CB samples. Study design and methods: To establish the most robust and efficient way to measure the Treg content of previously cryopreserved CB units, we compared the enumeration of Treg and CD3+ cells using flow cytometry and an epigenetic, DNA-based methodology. The two methods were assessed for their agreement, consistency and susceptibility to error when enumerating Treg and CD3+ cell numbers in both fresh and cryopreserved CB samples. Results: Epigenetic enumeration gave consistent and comparable results in both fresh and frozen CB samples. By contrast, assessment of Tregs and CD3+ cells by flow cytometry was only possible in fresh samples due to significant cell death following cryopreservation and thawing. Conclusion: Epigenetic assessment offers significant advantages over flow cytometry for analysing cryopreserved CB; similar cell numbers were observed both in fresh and frozen samples. Furthermore, multiple epigenetic assessments can be performed from DNA extracted from small cryopreserved CB segments; often the only CB sample available for clinical studies. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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38. The anticoagulant effects of ethyl pyruvate in whole blood samples.
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Haidl, Harald, Schlagenhauf, Axel, Krebs, Angelika, Plank, Harald, Wonisch, Willibald, Fengler, Vera, Fiegl, August, Hörl, Gerd, Koestenberger, Martin, Wagner, Thomas, Tafeit, Erwin, Cvirn, Gerhard, and Hallström, Seth
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ANTICOAGULANTS , *PARTIAL thromboplastin time , *BLOOD sampling , *SCANNING electron microscopy - Abstract
Background: Ethyl pyruvate (EP), the ethyl ester of pyruvate, has proven antiinflammatory and antioxidative properties. Additionally, anticoagulant properties have been suggested recently. EP, therefore, is a potentially antiatherosclerotic drug. We aimed to investigate whether EP possesses antiplatelet and anticoagulant properties particularly in the physiological environment of whole blood. Methods: We investigated the effects of increasing concentrations of EP on platelet function, on the course of clot development, and on standard coagulation times. Additionally, clot ultrastructure using scanning electron microscopy was analysed. Results: EP exerted significant antiplatelet actions: i) Impedance aggregometry amplitudes (11.7 ± 3.0 ohm, 0 μg/mL EP) dose dependently decreased (7.8 ± 3.1 ohm, 1000 μg/mL EP; -33.3%). ATP exocytosis (0.87 ± 0.24 nM, 0 μg/mL EP) measured by the luminiscent method dose-dependently decreased (0.56 ± 0.14 nM, 1000 μg/mL; -35.6%). ii) Closure times (104.4 ± 23.8 s, 0 μg/mL EP) using the Platelet function analyzer were dose-dependently prolonged (180.5 ± 82.5 s, 1000 μg/mL EP; +72.9%) using membranes coated with collagen/ADP. iii) Surface coverage (15.9 ± 5.1%, 0 μg/mL EP) dose-dependently decreased (9.0 ± 3.7%, 1000 μg/mL EP; -43.4%) using the Cone and Platelet analyzer. EP also exerted significant anticoagulant actions: Coagulation times (177.9 ± 37.8, 0 μg/mL EP) evaluated by means of thrombelastometry were dose-dependently prolonged (212.8 ± 57.7 s, 1000 μg/mL EP; +19.6%). Activated partial thromboplastin times (31.5 ± 1.8 s, 0 μg/mL EP) were dose-dependently prolonged (35.6 ± 2.3 s, 1000 μg/mL EP; +13.0%). Prothrombin times (0.94 ± 0.02 INR, 0 μg/mL EP) were dose-dependently prolonged (1.09 ± 0.04 INR, 1000 μg/mL EP; +16.0%). Conclusion: We found that EP possesses antiplatelet and anticoagulant properties in whole blood. Together with its proven anti-inflammatory and antioxidative properties, EP is a potentially antiatherogenic drug. [ABSTRACT FROM AUTHOR]
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- 2020
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39. Impact of three commonly used blood sampling techniques on the welfare of laboratory mice: Taking the animal's perspective.
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Meyer, Neele, Kröger, Mareike, Thümmler, Julia, Tietze, Lisa, Palme, Rupert, and Touma, Chadi
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LABORATORY mice , *BLOOD sampling , *SAMPLING (Process) , *LABORATORY techniques , *BLOOD collection , *ANIMAL welfare - Abstract
Laboratory mice are the most frequently used animals in biomedical research. In accordance with guidelines for humane handling, several blood sampling techniques have been established. While the effects of these procedures on blood quality and histological alterations at the sampling site are well studied, their impact on the animals' welfare has not been extensively investigated. Therefore, our study aimed to compare three commonly used blood sampling techniques regarding their effects on different indicators of animal welfare, including physiological and behavioural response stress parameters, including pain measures, home-cage behaviour and nest-building as well as exploratory activity and neophobia. Male C57BL/6J mice were subjected to a single blood collection from either the vena facialis, the retrobulbar sinus or the tail vessel, or were allocated to the respective control treatment. While all blood sampling techniques led to an acute increase in plasma corticosterone levels, the response was strongest in animals that underwent sampling from the vena facialis and the retrobulbar sinus. Similar results were observed when the time-course of adrenocortical activity was monitored via corticosterone metabolites from faecal samples. Blood collection from the vena facialis and the retrobulbar sinus also decreased exploration of novel stimuli, resulted in decreased nest-building activity and induced higher scores in the Mouse Grimace Scale. Moreover, locomotor activity and anxiety-related behaviour were strongly affected after facial vein bleeding. Interestingly, tail vessel bleeding only induced little alterations in the assessed physiological and behavioural parameters. Importantly, the observed effects in all treatment groups were no longer detectable after 24 hours, indicating only short-term impacts. Thus, by also taking the animal's perspective and comprehensively assessing the severity of the particular sampling procedures, the results of our study contribute to Refinement within the 3R concept and allow researchers to objectively select the most appropriate and welfare-friendly blood sampling technique for a given experiment. [ABSTRACT FROM AUTHOR]
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- 2020
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40. Evaluation of nucleosome concentrations in healthy dogs and dogs with cancer.
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Wilson-Robles, Heather, Miller, Tasha, Jarvis, Jill, Terrell, Jason, Dewsbury, Nathan, Kelly, Terry, Herzog, Marielle, Bygott, Thomas, Hardat, Nathalie, and Michel, Gaetan
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CHROMATIN , *DOGS , *CANCER , *CELL death , *BLOOD sampling - Abstract
Introduction: Nucleosomes consist of small fragments of DNA wrapped around a histone octamer core. Diseases such as cancer or inflammation lead to cell death, which causes fragmentation and release of nucleosomes into the blood. The Nu.Q™ technology measures circulating nucleosome levels and exploits the different compositions of cancer derived nucleosomes in blood to detect and identify cancer even at early stages. The objectives of this study are to identify the optimal sample type for the Nu.Q™ H3.1 assay and to determine if it can accurately detect nucleosomes in the blood of healthy canines as well as those with cancer. Materials and methods: Blood samples from healthy canine volunteers as well as dogs newly diagnosed with lymphoma were used. The blood was processed at a variety of times under a variety of conditions to determine the most reliable sample type and conditions, and to develop an appropriate processing strategy to ensure reliably accurate results. Results: Nucleosomes could be detected using a variety of sample collection and processing protocols. Nucleosome signals were highest in EDTA plasma and serum samples and most consistent in plasma. Samples should be processed within an hour of collection. Experiments showed that samples were able to withstand several freeze thaw cycles. Processing time and tcollection tube type did affect nucleosome detection levels. Finally, significantly elevated concentrations of nucleosomes were seen in a small cohort of dogs that had been newly diagnosed with lymphoma. Conclusions: When samples are collected and processed appropriately, the Nu.Q™ platform can reliably detect nucleosomes in the plasma of dogs. Further testing is underway to validate and optimize the Nu.Q™ platform for veterinary use. [ABSTRACT FROM AUTHOR]
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- 2020
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41. Comparison of loop-mediated isothermal amplification (LAMP) and PCR for the diagnosis of infection with Trypanosoma brucei ssp. in equids in The Gambia.
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Gummery, Lauren, Jallow, Saloum, Raftery, Alexandra G., Bennet, Euan, Rodgers, Jean, and Sutton, David G. M.
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TRYPANOSOMA brucei , *LAMPS , *TRYPANOSOMIASIS , *EQUIDAE , *BLOOD sampling , *TRYPANOSOMA - Abstract
Introduction: Infection of equids with Trypanosoma brucei (T. brucei) ssp. is of socioeconomic importance across sub-Saharan Africa as the disease often progresses to cause fatal meningoencephalitis. Loop-mediated isothermal amplification (LAMP) has been developed as a cost-effective molecular diagnostic test and is potentially applicable for use in field-based laboratories. Part I: Threshold levels for T. brucei ssp. detection by LAMP were determined using whole equine blood specimens spiked with known concentrations of parasites. Results were compared to OIE antemortem gold standard of T. brucei-PCR (TBR-PCR). Results I: Threshold for detection of T. brucei ssp. on extracted DNA from whole blood was 1 parasite/ml blood for LAMP and TBR-PCR, and there was excellent agreement (14/15) between tests at high (1 x 103/ml) concentrations of parasites. Detection threshold was 100 parasites/ml using LAMP on whole blood (LWB). Threshold for LWB improved to 10 parasites/ml with detergent included. Performance was excellent for LAMP at high (1 x 103/ml) concentrations of parasites (15/15, 100%) but was variable at lower concentrations. Agreement between tests was weak to moderate, with the highest for TBR-PCR and LAMP on DNA extracted from whole blood (Cohen's kappa 0.95, 95% CI 0.64–1.00). Part II: A prospective cross-sectional study of working equids meeting clinical criteria for trypanosomiasis was undertaken in The Gambia. LAMP was evaluated against subsequent TBR-PCR. Results II: Whole blood samples from 321 equids in The Gambia were processed under field conditions. There was weak agreement between LWB and TBR-PCR (Cohen's kappa 0.34, 95% CI 0.19–0.49) but excellent agreement when testing CSF (100% agreement on 6 samples). Conclusions: Findings support that LAMP is comparable to PCR when used on CSF samples in the field, an important tool for clinical decision making. Results suggest repeatability is low in animals with low parasitaemia. Negative samples should be interpreted in the context of clinical presentation. [ABSTRACT FROM AUTHOR]
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- 2020
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42. Early conversion to a CNI-free immunosuppression with SRL after renal transplantation—Long-term follow-up of a multicenter trial.
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Andrassy, Joachim, Guba, Markus, Habicht, Antje, Fischereder, Michael, Pratschke, Johann, Pascher, Andreas, Heller, Katharina M., Banas, Bernhard, Hakenberg, Oliver, Vogel, Thomas, Meiser, Bruno, Dick, Andrea, Werner, Jens, and Kauke, Teresa
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IMMUNOSUPPRESSION , *BLOOD sampling - Abstract
Introduction: Early conversion to a CNI-free immunosuppression with SRL was associated with an improved 1- and 3- yr renal function as compared with a CsA-based regimen in the SMART-Trial. Mixed results were reported on the occurrence of donor specific antibodies under mTOR-Is. Here, we present long-term results of the SMART-Trial. Methods and materials: N = 71 from 6 centers (n = 38 SRL and n = 33 CsA) of the original SMART-Trial (ITT n = 140) were enrolled in this observational, non-interventional extension study to collect retrospectively and prospectively follow-up data for the interval since baseline. Primary objective was the development of dnDSA. Blood samples were collected on average 8.7 years after transplantation. Results: Development of dnDSA was not different (SRL 5/38, 13.2% vs. CsA 9/33, 27.3%; P = 0.097). GFR remained improved under SRL with 64.37 ml/min/1.73m2 vs. 53.19 ml/min/1.73m2 (p = 0.044). Patient survival did not differ between groups at 10 years. There was a trend towards a reduced graft failure rate (11.6% SRL vs. 23.9% CsA, p = 0.064) and less tumors under SRL (2.6% SRL vs. 15.2% CsA, p = 0.09). Conclusions: An early conversion to SRL did not result in an increased incidence of dnDSA nor increased long-term risk for the recipient. Transplant function remains improved with benefits for the graft survival. [ABSTRACT FROM AUTHOR]
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- 2020
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43. HIV testing, care and viral suppression among men who have sex with men and transgender individuals in Johannesburg, South Africa.
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Fearon, Elizabeth, Tenza, Siyanda, Mokoena, Cecilia, Moodley, Kerushini, Smith, Adrian D., Bourne, Adam, Weatherburn, Peter, and Palanee-Phillips, Thesla
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HIV , *POISSON regression , *VIRAL load , *HIV-positive persons , *BLOOD sampling - Abstract
Introduction: Men who have sex with men and transgender individuals (MSM/TG) carry a disproportionately high burden of HIV, including in South Africa. However, there are few empirical population-representative estimates of viral suppression and the HIV care cascade including HIV testing among this population, nor of factors associated with these outcomes. Methods: We conducted a respondent driven sampling (RDS) survey among 301 MSM/TG in Johannesburg in 2017. Participants gave blood samples for HIV testing and viral load. Participants self-completed a survey including sociodemographics, HIV testing history, and engagement in care. We calculated RDS-II weighted estimates of the percentage of HIV-negative MSM/TG reporting HIV testing in the previous 6 months, their testing experience and preferences. Among those HIV-positive, we estimated the percentage status-aware, on ART, and virally suppressed (<50 viral copies/ml plasma). We conducted RDS-weighted robust Poisson regression to obtain weighted prevalence ratios of factors associated with 1) HIV testing among those HIV-negative; and 2) viral suppression among those HIV-positive. Results: There were 118/300 HIV-positive MSM/TG, (37.5%). Of the HIV-negative MSM/TG, 61.5% reported that they had tested for HIV in the previous 6 months, which was associated with selling sex to men (Prevalence Ratio = 1.67, 95% CI 1.36–2.05). There were 76/118 HIV-positive MSM/TG (56.5%) who reported having previously tested positive for HIV and 39/118 (30.0%) who reported current ART. There were 58/118 HIV-positive MSM/TG with viral loads <50 copies/ml plasma (46.9%). Viral suppression was associated with older age (adjusted PR = 1.03, 95% CI 1.00–1.06 for each year), neighbourhood, and having bought sex from men (adjusted PR = 1.53, 95% CI 1.12–2.08). Conclusions: HIV prevalence was very high. Viral suppression among those HIV-positive was similar to the general male population in South Africa, but remains far short of national and international targets. A majority of HIV-negative MSM/TG had HIV tested in the previous 6 months, though there is room for improvement. [ABSTRACT FROM AUTHOR]
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- 2020
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44. Akwa Ibom AIDS indicator survey: Key findings and lessons learnt.
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Adedokun, Oluwasanmi, Badru, Titilope, Khamofu, Hadiza, Negedu-Momoh, Olubunmi Ruth, Iwara, Emem, Agbakwuru, Chinedu, Atobatele, Akinyemi, Merrigan, Mike, Ukpong, Dominic, Nzelu, Charles, Ashefor, Gregory, Pandey, Satish Raj, and Torpey, Kwasi
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HIV-positive children , *HIV prevention , *BLOOD collection , *VIRAL load , *HOUSEHOLDS , *BLOOD sampling , *AIDS - Abstract
Background: The burden of HIV/AIDS epidemic is huge, but this varies widely by population in Nigeria. Data that could be used to guide the scale up of HIV prevention and control strategies has significant gaps. The study sought to estimate the prevalence of HIV and its associated determinants in Akwa Ibom state. Methods: Akwa Ibom AIDS Indicator Survey (AKAIS) is a population based cross-sectional survey, with a two-stage probability sampling. The survey had both behavioural and biological components. Tablet-based questionnaire was used to collect data on participant's household information, demographics, socio-economic, and behavioral risk factors associated with HIV; while the biological component involved collection of venous blood samples for participants who were over 19months. For children aged 18months on less, capillary blood from finger prick sample was used. Participants were tested for HIV. Other biomarker tests for HIV positive participants included CD4, HIV-1 RNA viral load and incidence assays. Results: In all 15,609 people (8,963 adults aged 15 years and older (55% females), 6,646 individuals less than 15 years (51% males), from 4,313 households, participated in AKAIS. Overall, 2.8% (423 persons; 422 HIV-1 and 1 HIV-2) were found to be HIV positive. HIV prevalence was 4.8% in adults (15 years and above) and 0.4% in pediatric (< = 14 years) participants. HIV prevalence was significantly higher in females (5.6%) than males (3.7%) aged 15 years and older (p <0.001). Overall HIV incidence was 0.41% Conclusions: HIV prevalence among adults was 4.8% with an overall incidence of 0.41%. These estimates are essential to inform strategic control and prevention of HIV epidemic in Akwa Ibom state targeting the affected populations. [ABSTRACT FROM AUTHOR]
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- 2020
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45. Comparison of real time and malachite-green based loop-mediated isothermal amplification assays for the detection of Plasmodium vivax and P. falciparum.
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Barazorda, Keare A., Salas, Carola J., Bishop, Danett K., Lucchi, Naomi, and Valdivia, Hugo O.
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PLASMODIUM vivax , *PLASMODIUM falciparum , *TURNAROUND time , *PLASMODIUM , *MALARIA , *BLOOD sampling - Abstract
The current context of malaria elimination requires urgent development and implementation of highly sensitive and specific methods for prompt detection and treatment of malaria parasites. Such methods should overcome current delays in diagnosis, allow the detection of low-density infections and address the difficulties in accessing remote endemic communities. In this study, we assessed the performance of the RealAmp and malachite-green loop mediated isothermal amplification (MG-LAMP) methodologies, using microscopy and conventional nested-PCR as reference techniques. Both LAMP techniques were performed for Plasmodium genus, P. falciparum, and P. vivax identification using 136 whole blood samples collected from three communities located in the Peruvian Amazon basin. Turnaround time and costs of performing the LAMP assays were estimated and compared to that of microscopy and nested-PCR. Using nested-PCR as reference standard, we calculated the sensitivity, specificity and 95% confidence interval (CI) for all methods. RealAmp had a sensitivity of 92% (95% CI: 85–96.5%) and specificity of 100% (95% CI: 89.1–100%) for species detection; sensitivity and specificity of MG-LAMP were 94% (95% CI: 87.5–97.8%) and 100% (89.1–100%), respectively. Whereas microscopy showed 88.1% sensitivity (95% CI: 80.2–93.7%) and 100% specificity (95%: 89.1–100%). The turnaround time and costs of performing the LAMP assays were lower compared to those associated with nested-PCR but higher than those associated with microscopy. The two LAMP assays were shown to be more sensitive and simple to implement than microscopy. Both LAMP methodologies could be used as large-scale screening tests, but the MG-LAMP assay uses a simple, portable heat-block while the RealAmp requires a RealAmp machine or a real-time PCR machine. This makes the MG-LAMP an appropriate choice for malaria surveillance studies in endemic sites. Use of LAMP tests in active case detection of Plasmodium parasites could help to detect positive malaria cases early. [ABSTRACT FROM AUTHOR]
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- 2020
- Full Text
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46. UltraPrep is a scalable, cost-effective, bead-based method for purifying cell-free DNA.
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Raymond, Christopher K., Raymond, Fenella C., and Hill, Kay
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DNA , *BLOOD collection , *MOLECULAR weights , *BLOOD sampling - Abstract
UltraPrep is an open-source, two-step method for purification of cell-free DNA that entails extraction of total DNA followed by size-selective enrichment of the smaller fragments that are characteristic of DNA originating from fragmentation between nucleosome. The advantages of the two related protocols that are described are that they can easily accommodate a wide range of sample input volumes, they rely on simple, magnetic bead-based technology, the yields of cfDNA are directly comparable to the most popular methods for cfDNA purification, and they dramatically reduce the cost of cfDNA isolation relative to currently available commercial methods. We provide a framework for physical and molecular quality analysis of purified cfDNA and demonstrate that the cfDNA generated by UltraPrep meets or exceeds the quality metrics of the most commonly used procedure. In addition, our method removes high molecular weight genomic DNA (hmwgDNA) that can interfere with downstream assay results, thereby addressing one of the primary concerns for preanalytical collection of blood samples. [ABSTRACT FROM AUTHOR]
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- 2020
- Full Text
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47. Plasma Galectin-3 predicts deleterious vascular dysfunction affecting post-myocardial infarction patients: An explanatory study.
- Author
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Huttin, Olivier, Mandry, Damien, Popovic, Batric, Rossignol, Patrick, Odille, Freddy, Micard, Emilien, Lamiral, Zohra, Zannad, Faïez, Girerd, Nicolas, and Marie, Pierre-Yves
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GALECTINS , *INFARCTION , *VASCULAR resistance , *BLOOD sampling , *MYOCARDIAL infarction , *CARDIAC amyloidosis - Abstract
Objectives: In a previous analysis of a post-myocardial infarction (MI) cohort, abnormally high systemic vascular resistances (SVR) were shown to be frequently revealed by MRI during the healing period, independently of MI severity, giving evidence of vascular dysfunction and limiting further recovery of cardiac function. The present ancillary and exploratory analysis of the same cohort was aimed at characterizing those patients suffering from high SVR remotely from MI with a large a panel of cardiovascular MRI parameters and blood biomarkers. Methods: MRI and blood sampling were performed 2–4 days after a reperfused MI and 6 months thereafter in 121 patients. SVR were monitored with a phase-contrast MRI sequence and patients with abnormally high SVR at 6-months were characterized through MRI parameters and blood biomarkers, including Galectin-3, an indicator of cardiovascular inflammation and fibrosis after MI. SVR were normal at 6-months in 90 patients (SVR-) and abnormally high in 31 among whom 21 already had high SVR at the acute phase (SVR++) while 10 did not (SVR+). Results: When compared with SVR-, both SVR+ and SVR++ exhibited lower recovery in cardiac function from baseline to 6-months, while baseline levels of Galectin-3 were significantly different in both SVR+ (median: 14.4 (interquartile range: 12.3–16.7) ng.mL-1) and SVR++ (13.0 (11.7–19.4) ng.mL-1) compared to SVR- (11.7 (9.8–13.5) ng.mL-1, both p < 0.05). Plasma Galectin-3 was an independent baseline predictor of high SVR at 6-months (p = 0.002), together with the baseline levels of SVR and left ventricular end-diastolic volume, whereas indices of MI severity and left ventricular function were not. In conclusion, plasma Galectin-3 predicts a deleterious vascular dysfunction affecting post-MI patients, an observation that could lead to consider new therapeutic targets if confirmed through dedicated prospective studies. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
48. CCNB2 and AURKA overexpression may cause atypical mitosis in Japanese cortisol-producing adrenocortical carcinoma with TP53 somatic variant.
- Author
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Ikeya, Akira, Nakashima, Mitsuko, Yamashita, Miho, Kakizawa, Keisuke, Okawa, Yuta, Saitsu, Hirotomo, Sasaki, Shigekazu, Sasano, Hironobu, Suda, Takafumi, and Oki, Yutaka
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MITOSIS , *CARCINOMA , *NUCLEOTIDE sequencing , *HYDROCORTISONE , *BLOOD sampling , *DNA , *JAPANESE people - Abstract
Background: Many genomic analyses of cortisol-producing adrenocortical carcinoma (ACC) have been reported, but very few have come from East Asia. The first objective of this study is to verify the genetic difference with the previous reports by analyzing targeted deep sequencing of 7 Japanese ACC cases using next-generation sequencing (NGS). The second objective is to compare the somatic variant findings identified by NGS analysis with clinical and pathological findings, aiming to acquire new knowledge about the factors that contribute to the poor prognosis of ACC and to find new targets for the treatment of ACC. Method: DNA was extracted from ACC tissue of seven patients and two reference blood samples. Targeted deep sequencing was performed using the MiSeq system for 12 genes, and the obtained results were analyzed using MuTect2. The hypothesis was obtained by integrating the somatic variant findings with clinical and pathological data, and it was further verified using The Cancer Genome Atlas (TCGA) dataset for ACC. Results: Six possible pathogenic and one uncertain significance somatic variants including a novel PRKAR1A (NM_002734.4):c.545C>A (p.T182K) variant were found in five of seven cases. By integrating these data with pathological findings, we hypothesized that cases with TP53 variants were more likely to show atypical mitotic figures. Using TCGA dataset, we found that atypical mitotic figures were associated with TP53 somatic variant, and mRNA expression of CCNB2 and AURKA was significantly high in TP53 mutated cases and atypical mitotic figure cases. Conclusion: We believe this is the first report that discusses the relationship between atypical mitotic figures and TP53 somatic variant in ACC. We presumed that overexpression of CCNB2 and AURKA mRNA may cause atypical mitosis in TP53 somatic mutated cases. Because AURKA is highly expressed in atypical mitotic cases, it may be an appropriate indicator for AURKA inhibitors. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
49. Dried blood spot self-sampling at home is a feasible technique for hepatitis C RNA detection.
- Author
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Prinsenberg, Tamara, Rebers, Sjoerd, Boyd, Anders, Zuure, Freke, Prins, Maria, van der Valk, Marc, and Schinkel, Janke
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HEPATITIS C , *RNA , *VIRAL load , *BLOOD , *BLOOD sampling - Abstract
To facilitate HCV diagnosis, we developed an HCV-RNA testing service, which involved home-sampled dried blood spots (DBS). The main objective of this study was to evaluate the feasibility of self-sampling at home. Furthermore, to optimise the processing of DBS samples for RNA detection, we evaluated two elution buffers: phosphate-buffered saline (PBS) and L6-buffer. 27 HCV-RNA and 12 HIV-1 RNA positive patients were included. Laboratory spotted DBS (LabDBS) were made by a technician from blood samples drawn at inclusion. Patients received a DBS home-sampling kit and were requested to return their self-sampled DBS (ssDBS) by mail. We compared the RNA load of PBS and L6-eluted labDBS, and of L6-eluted ssDBS, L6-eluted labDBS and plasma. LabDBS load measurements were repeated after 7–13 and 14–21 days to evaluate RNA stability. All 39 plasma samples provided quantifiable RNA loads. In 1/39 labDBS sample, RNA could not be detected (plasma HCV load: 2.98 log10 IU/ml). L6-eluted samples gave a 0.7 log10 and 0.6 log10 higher viral load for HCV and HIV-1 respectively, compared to PBS-eluted samples. Strong correlations were found between labDBS and ssDBS HCV RNA (r = 0.833; mean difference 0.3 log10 IU/mL) and HIV-1 RNA results (r = 0.857; mean difference 0.1 log10 copies/mL). Correlations between labDBS and plasma values were high for HCV (r = 0.958) and HIV-1 (r = 0.844). RNA loads in DBS remained stable over 21 days. Our study demonstrates that self-sampling dried blood spots at home is a feasible strategy for the detection of HCV and HIV-1 RNA. This could facilitate one-step diagnostics and treatment monitoring in communities with high HCV prevalence. [ABSTRACT FROM AUTHOR]
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- 2020
- Full Text
- View/download PDF
50. The impact of vitamin D3 intake on inflammatory markers in multiple sclerosis patients and their first-degree relatives.
- Author
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Hashemi, Reza, Hosseini-Asl, Seyed Saeed, Arefhosseini, Seyed Rafie, and Morshedi, Mohammad
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FAMILY history (Medicine) , *MULTIPLE sclerosis , *VITAMINS , *CALCITRIOL , *NATALIZUMAB , *BLOOD sampling , *MESSENGER RNA - Abstract
Background & aims: In our previous study, a Seesaw model was proposed for the fluctuation of crucial anti- (IL-10) and pro-inflammatory (Il-6 & IL-17A) cytokines through vitamin D3. In this paper, however, it is intended to extend the mentioned model by assessing the expression mRNA levels of IL-27 and TGF-β1 as well as the changes of plasma levels of IL-27, TGF-β1, IL-17A, IL-10, and IL-6 after treatment by vitamin D3. Method: Venous blood samples were drawn from Healthy Participants (HP, n = 25) and First-Degree Relative Participants (FDRP, n = 25) as control groups and Multiple Sclerosis Participants (MSP, n = 25) before and after eight weeks of supplementation with 50000 IU vitamin D3. The mRNA expression and plasma concentrations were gauged by using Real-Time PCR and ELISA assay, respectively. Results: The mRNA surfaces of IL-27, as well as TGF-β1, were up-regulated. However, the plasma levels of TGF-β1, IL-17A, and IL-6 were significantly different among the three groups. In addition, the plasma levels of IL-27, TGF-β1, IL-10, IL-17A, and IL-6 significantly changed following the administration of vitamin D3. Conclusion: The findings of this paper illustrate that anti-inflammatory cytokines could have a key role in immunomodulatory functions due to their anti-inflammatory functions. To conclude, this might contribute to preventing the pathophysiological process of MS. Also, the proposed model could be used as a preventive way on disposed people to multiple sclerosis, particularly in first degree relatives of these patients. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
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