scholarly journals Enrichment of leukocytes in peripheral blood using 3D printed tubes

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0254615
Author(s):  
Li-fang Guo ◽  
Liu Wang ◽  
Sai Ren ◽  
Ning Su ◽  
Kun Wei ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Basic Research ◽  
Buffy Coat ◽  
Cellular Damage ◽  
Label Free ◽  
Recovery Fraction ◽  
Phase Cycle ◽  
Centrifugation Step ◽  
3D Printed ◽  
New Procedures

Leukocytes have an essential role in patient clinical trajectories and progression. Traditional methods of leukocyte enrichment have many significant limitations for current applications. It is demonstrated a novel 3D printing leukocyte sorting accumulator that combines with centrifugation to ensure label-free initial leukocyte enrichment based on cell density and size. The internal structure of leukocyte sorting accumulator (revealed here in a new design, leukocyte sorting accumulator-3, upgraded from earlier models), optimizes localization of the buffy coat fraction and the length of the period allocated for a second centrifugation step to deliver a higher recovery of buffy coats than earlier models. Established methodological parameters were evaluated for reliability by calculating leukocyte recovery rates and erythrocyte depletion rates by both pushing and pulling methods of cell displacement. Results indicate that leukocyte sorting accumulator-3 achieves a mean leukocytes recovery fraction of 96.2 ± 2.38% by the pushing method of layer displacement. By the pulling method, the leukocyte sorting accumulator-3 yield a mean leukocytes recovery fraction of 94.4 ± 0.8%. New procedures for preliminary enrichment of leukocytes from peripheral blood that avoid cellular damage, as well as avert metabolic and phase cycle intervention, are required as the first step in many modern clinical and basic research assays.

scholarly journals Evaluation of qPCR on blood and skin microbiopsies, peripheral blood buffy coat smear, and urine antigen ELISA for diagnosis and test of cure for visceral leishmaniasis in HIV-coinfected patients in India: a prospective cohort study

BMJ Open ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. e042519
Author(s):  
Sophie I Owen ◽  
Sakib Burza ◽  
Shiril Kumar ◽  
Neena Verma ◽  
Raman Mahajan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Hiv Infection ◽  
Peripheral Blood ◽  
Medical Science ◽  
Bone Marrow Aspiration ◽  
Buffy Coat ◽  
Tropical Medicine ◽  
Urine Antigen ◽  
Antigen Elisa

IntroductionHIV coinfection presents a challenge for diagnosis of visceral leishmaniasis (VL). Invasive splenic or bone marrow aspiration with microscopic visualisation of Leishmania parasites remains the gold standard for diagnosis of VL in HIV-coinfected patients. Furthermore, a test of cure by splenic or bone marrow aspiration is required as patients with VL-HIV infection are at a high risk of treatment failure. However, there remain financial, implementation and safety costs to these invasive techniques which severely limit their use under field conditions.Methods and analysisWe aim to evaluate blood and skin qPCR, peripheral blood buffy coat smear microscopy and urine antigen ELISA as non-invasive or minimally invasive alternatives for diagnosis and post-treatment test of cure for VL in HIV-coinfected patients in India, using a sample of 91 patients with parasitologically confirmed symptomatic VL-HIV infection.Ethics and disseminationEthical approval for this study has been granted by The Liverpool School of Tropical Medicine, The Institute of Tropical Medicine in Antwerp, the University of Antwerp and the Rajendra Memorial Research Institute of Medical Science in Patna. Any future publications will be published in open access journals.Trial registration numberCTRI/2019/03/017908.

scholarly journals Current aspects of pathogenesis and prophylaxis of pelvic adhesions

2020 ◽  
Vol 14 (4) ◽  
pp. 523-533
Author(s):  
G. A. Puchkina ◽  
A. N. Sulima
Keyword(s):  
Open Access ◽  
Peripheral Blood ◽  
Peritoneal Fluid ◽  
Preventive Measures ◽  
Basic Research ◽  
Published Data ◽  
Drug Form ◽  
Peritoneal Adhesions ◽  
Pelvic Organs ◽  
Open Access Journals

Introduction. Adhesive process of the pelvic organs is a pressing issue for operative gynecology that does not allow to consider results of therapeutic and preventive measures as sufficient.Aim: to analyze published data regarding contemporary aspects of etiopathogenesis of the adhesive process in pelvic organs as well as methods of its prevention.Materials and Methods. The literature sources retrieved from electronic databases PubMed, Embase, Medline, Ovid HealthSTAR, Cochrane, Google Scholar, eLibrary, CyberLeninka as well as scientific articles published in peer-reviewed open access journals over the last 30 years, including basic research in the field have been analyzed. While searching, the following keywords and their combinations in Russian and English were used: "adhesive process of the pelvic organs", "pathogenesis of the adhesive process", "prevention of the adhesive process", "gynecology", "pelvic adhesions", "pathogenesis of adhesions", "аdhesion prophylaxis", "gynecology".Results. The current aspects of the etiology and pathogenesis for adhesive process have been summarized. Existing adhesion classifications are presented. The proposed methods for preventing formation of peritoneal adhesions are described exerting most prominent effectiveness as well as describing the properties and characteristics according to the application method, the composition of contained substances and drug form. A phenotype profile of peripheral blood and peritoneal fluid lymphocytes from patients with adhesive process remains debated.Conclusion. A need to further examine formation of peritoneal adhesions at molecular and cellular levels for developing a comprehensive pathogenetically substantiated method to prevent and treat adhesions of the pelvic organs is in demand.

scholarly journals Cellular effects of gliotoxin: Evaluation of a proteomic, isotope-based method to detect reactive cysteines

2021 ◽  
Author(s):  
◽  
Sven Sondhauss
Keyword(s):  
Large Scale ◽  
Thiol Group ◽  
Disulfide Bridge ◽  
Reversed Phase ◽  
Cellular Damage ◽  
Full Potential ◽  
Label Free ◽  
Cysteine Modification ◽  
Affinity Tag ◽  
Cell Vitality

<p>Cysteinyl residues in proteins are important for many cellular processes and unregulated modification of the cysteine thiol group can have negative effects on cell vitality and viability. In this thesis, the potential for use of the isotope coded affinity tag (ICAT) method for detection of cysteine modification has been investigated. ICAT reagents label free cysteine thiols. The aim of this study was to use HL-60 cells treated with gliotoxin, a fungal metabolite with a reactive disulfide bridge, as a system to evaluate the performance of ICAT for identification of cysteine modification in a whole cell proteome. Gliotoxin has antimicrobial, antitumor, immunosuppressive and cytotoxic properties that have been related to cysteine modification in proteins. Cellular assays including viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, cell cycle analysis, and measurement of reactive oxygen species using dichlorofluorescin diacetate were used to establish conditions for measuring the effects of gliotoxin on HL-60 cells prior to large-scale cellular damage. Cells exposed to gliotoxin and control cells were then labeled with ICAT reagents and analysed by offline reversed phase liquid chromatography followed by matrix-assisted laser desorption/ionization tandem mass spectrometry. The pilot results identified tubulin, glyceraldehyde-3-phosphate dehydrogenase and peptidyl-prolyl cis-trans isomerase as putative targets of gliotoxin. Additionally, this study showed that ICAT can be used to detect modified cysteines from a highly complex sample, but further optimization is needed to unlock the full potential for detection of cysteine modification in complex samples.</p>

Enzootic Bovine Leukemia: C-type Virus in a Cow with Acute Leukemia and in Cattle with Persistent Lymphocytosis from a Dairy Herd.

1975 ◽  
Vol 61 (3) ◽  
pp. 261-270 ◽  
Author(s):  
Luigi Rampichini ◽  
Fernando L. De Castro Portugal ◽  
Maurizio Severini ◽  
Domenico Rutili
Keyword(s):  
Peripheral Blood ◽  
Dairy Herd ◽  
Buffy Coat ◽  
Acute Lymphatic Leukemia ◽  
Persistent Lymphocytosis ◽  
Ultrastructural Studies ◽  
Milk Samples ◽  
Lymphatic Leukemia ◽  
Hematologic Examination ◽  
Microscopy Examination

A case of acute lymphatic leukemia in a 7-month-pregnant Italian Friesian cow, aged about 6 years and belonging to a small dairy herd of 11 cows is described. Hematologic, histopatologic and ultrastructural examinations were performed, the ultrastructural studies were confined to the lymphnodes, thymus and buffy-coat cultures from peripheral blood. The remaining animals were subjected to hematologic examination and electron microscopy examination of buffy-coat cultures from peripheral blood. C-type particles were found in phytohemagglutinin (PHA)-treated and untreated buffy-coat cultures from the leukemic cow and from the animals with persistent lymphocytosis. C-type particles were also found in milk samples from 3 cows with persistent lymphocytosis.

scholarly journals QBC® for the diagnosis of human and canine american visceral leishmaniasis: preliminary data

2001 ◽  
Vol 34 (6) ◽  
pp. 577-581 ◽  
Author(s):  
Daniel B. Liarte ◽  
Ivete L. Mendonça ◽  
Francisco C.O. Luz ◽  
Elza A.S. de Abreu ◽  
Gustavo W.S. Mello ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Preliminary Data ◽  
Peripheral Blood ◽  
Buffy Coat ◽  
Promising Method ◽  
Blood Parasites ◽  
Leishmania Chagasi ◽  
Asymptomatic Carriers ◽  
Fluorescent Method

"Quantitative Buffy Coat" (QBC®) is a direct and fast fluorescent method used for the identification of blood parasites. Since Leishmania chagasi circulates in blood, we decided to test it in American visceral leishmaniasis (AVL). Bone marrow (BM) and peripheral blood (PB) of 49 persons and PB of 31 dogs were analyzed. QBC® was positive in BM of 11/11 patients with AVL and in 1/6 patients with other diseases. Amastigotes were identified in PB of 18/22 patients with AVL and in none without AVL. The test was positive in 30 out of the 31 seropositive dogs and in 28/28 dogs with Leishmania identified in other tissues. QBC® is a promising method for diagnosis of human AVL, and possibly for the exam of PB of patients with AVL/AIDS, for the control of the cure and for the identification of asymptomatic carriers. Because it is fast and easy to collect and execute, QBC® should be evaluated for programs of reservoir control.

scholarly journals Nanophotonic Metasurfaces for Biosensing and Imaging

2019 ◽  
Vol 215 ◽  
pp. 12001
Author(s):  
Hatice Altug
Keyword(s):  
Scale Effects ◽  
Point Of Care ◽  
Optical Computing ◽  
Basic Research ◽  
Critical Issue ◽  
Label Free ◽  
Disruptive Technologies ◽  
Disease Diagnostics ◽  
Wide Range ◽  
Novel Devices

Nanophotonics excels at confining light into nanoscale optical mode volumes and generating dramatically enhanced light matter interactions. These unique aspects have been unveiling a plethora of fundamentally new optical phenomena, yet a critical issue ahead for nanophotonics is the development of novel devices and applications that can take advantage of these nano-scale effects. It is expected that nanophotonics will lead to disruptive technologies in energy harvesting, quantum and integrated photonics, optical computing and including biosensing. To this end, our research is focused on the application of nanophotonics to introduce powerful biosensors that can have impact on a wide range of areas including basic research in life sciences, early disease diagnostics, safety and point-of-care testing. In particular, we exploit nanophotonics and its integration with microfluidics to address key challenges of current biosensors and develop devices that can enable label-free, ultra-sensitive, multiplexed, rapid and real-time measurements on biomolecules, pathogens and living systems. In this talk I will present some of our recent work on nanophotonic meta surfaces for biosensing and bioimaging as well as their applications in real-world settings.

scholarly journals Bioimprint Mediated Label‐Free Isolation of Pancreatic Tumor Cells from a Healthy Peripheral Blood Cell Population

2020 ◽  
Vol 4 (11) ◽  
pp. 2000054
Author(s):  
Marie Pelle ◽  
Anupam A. K. Das ◽  
Leigh A. Madden ◽  
Vesselin N. Paunov
Keyword(s):  
Blood Cell ◽  
Tumor Cells ◽  
Cell Population ◽  
Peripheral Blood ◽  
Pancreatic Tumor ◽  
Peripheral Blood Cell ◽  
Label Free

scholarly journals Staphylococcus aureus Induces Hypoxia and Cellular Damage in Porcine Dermal Explants

2015 ◽  
Vol 83 (6) ◽  
pp. 2531-2541 ◽  
Author(s):  
Abdul G. Lone ◽  
Erhan Atci ◽  
Ryan Renslow ◽  
Haluk Beyenal ◽  
Susan Noh ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Death ◽  
Effective Diffusion ◽  
Oxygen Demand ◽  
Shotgun Proteomics ◽  
Cellular Damage ◽  
Label Free ◽  
Content Type ◽  
Dermal Tissue ◽  
The Difference

We developed a porcine dermal explant model to determine the extent to whichStaphylococcus aureusbiofilm communities deplete oxygen, change pH, and produce damage in underlying tissue. Microelectrode measurements demonstrated that dissolved oxygen (DO) in biofilm-free dermal tissue was 4.45 ± 1.17 mg/liter, while DO levels for biofilm-infected tissue declined sharply from the surface, with no measurable oxygen detectable in the underlying dermal tissue. Magnetic resonance imaging demonstrated that biofilm-free dermal tissue had a significantly lower relative effective diffusion coefficient (0.26 ± 0.09 to 0.30 ± 0.12) than biofilm-infected dermal tissue (0.40 ± 0.12 to 0.48 ± 0.12;P< 0.0001). Thus, the difference in DO level was attributable to biofilm-induced oxygen demand rather than changes in oxygen diffusivity. Microelectrode measures showed that pH within biofilm-infected explants was more alkaline than in biofilm-free explants (8.0 ± 0.17 versus 7.5 ± 0.15, respectively;P< 0.002). Cellular and nuclear details were lost in the infected explants, consistent with cell death. Quantitative label-free shotgun proteomics demonstrated that both proapoptotic programmed cell death protein 5 and antiapoptotic macrophage migration inhibitory factor accumulated in the infected-explant spent medium, compared with uninfected-explant spent media (1,351-fold and 58-fold, respectively), consistent with the cooccurrence of apoptosis and necrosis in the explants. Biofilm-origin proteins reflected an extracellular matrix-adapted lifestyle ofS. aureus. S. aureusbiofilms deplete oxygen, increase pH, and induce cell death, all factors that contribute to impede wound healing.

Routine Automated Processing of Cord Blood Units Using the Sepax Cell Processing System.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3646-3646
Author(s):  
Safa Karandish ◽  
Nery Berrios ◽  
Sufira Kiran ◽  
Charisse Ayuste ◽  
Toby Hamblin ◽  
...  
Keyword(s):  
Cord Blood ◽  
Processing Time ◽  
Processing System ◽  
Buffy Coat ◽  
Bar Code ◽  
Cell Processing ◽  
Wide Range ◽  
Significant Difference ◽  
Centrifugation Step ◽  
Cryoprotectant Solution

Abstract We recently incorporated the use of the automated Sepax Cell Processing System (Biosafe SA, Switzerland) for red cell and volume reduction of cord blood units (CBU) before cryopreservation. Now that we have been routinely using this new technique in the laboratory for about six months, we decided to compare the results of this method with our standard manual processing method (Rubinstein, et al, PNAS1995,; 92: 10119–22). For both methods, hespan is added to the cells at final concentration of 20% (v/v). With the Sepax system, after addition of hespan, the cell bag is connected to the Sepax tubing set with the final freeze bag pre-attached to the set. After completion of the automated procedure, buffy coat is collected in the final freeze bag. Cryoprotectant solution is then added directly to the freeze bag. In the manual method, buffy coat and white cell rich plasma layer is collected after first centrifugation step and the white cells are separated from the plasma after the second centrifugation step. Cryoprotectant solution is then added to the cells before transfer to the final freeze bag for cryopreservation. The following are summary of results for each method: Table 1 Manual (n=1160) Sepax (n=311) Pre-Processing volume (ml) 107±30 114±28 Pre-Processing TNC (×10e6) 1196±577 1315±519 TNC Recovery (%) 80±8 83±8 TNC Viability (%) (7-AAD) 97±3 98±3 Total CD34 (×10e6) 4.3±4 4.9±3.6 Total DFU (×10e6) 70±0.9 61.5±20 Post Processing RBC Volume (ml) 9±2 7.3±2 Total Processing Time (including Setup) 60 minutes 30 minutes It is important to note that there was not a significant difference in TNC Recovery over a wide range of Pre-Processing Volume (66–206ml) or Pre-Processing TNC (440 – 3559×106). Since the Sepax device is an automated procedure, issues could arise (i.e. short term loss of electrical power) that would require us to reprocess the CBU before cryopreservation. The Sepax system allows for recovery and reprocessing of the cell using the ‘Purge mode’. We used 5 CBU units to evaluate TNC recovery and viability after purging the cells once and reprocessing. Table 2 TNC Recovery (%) TNC Viability (%) First Buffy Coat 85±5 98±0.9 Post Purge and reprocessing 76±5 98±0.9 Although the TNC recovery was lower after the second procedure, it was still within acceptable limits and the viability of the cells had not changed. These data demonstrates that both methods are equivalent with respect to cell recovery. However, the Sepax System substantially reduces processing time and hands-on operator intervention. Additionally system provides, closed-system processing, bar code reading capability and run data print-out suitable for GMP manufacturing settings.

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