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Hematology - Science topic

A forum to address questions regarding hematology including the study of blood, the blood-forming organs, and blood diseases.
Questions related to Hematology
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Which prophylactic antibacterial therapy apply you to thecpatients with acute leukemia during an induction therapy or to the hematologic patients during a high-dose Chemotherapy and autologous stem cell transplantation?
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Commonly used combination is ciprofloxacin (anti-bacterial), an azole (itraconazole, voriconazole...) or ambisome (anti-fungal) and aciclovir (anti-viral).
Siva
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I have used Mindray BC-3000plus Hematology analyzer for research purpose.
Some information about may machine, Mindray BC 3000 Plus Fully Automatic Hematology Analyzer - 3 Part. Operation Mode-Fully Automatic, Brand-Mindray, differential Type-3-Part, Number of Parameters-19, Chamber Type-Double Chamber, Instrument Type-3 Part Hematology Analyzer. of running samples it displays error message gives error (background abnormal , WBC clog &RBC clog, when it aspirate sample or blank there red dots under screen but WBC and RBC remain zero it doesn't up count .finally it display preparing count time similarly WBC and RBC remain zero, empty histogram displayed) I was checked all setups , Maintained stepwise based on manual instruction ,cleaned all valves ,change WBC bath and I checked all setting I found that WBC reference count time and absolute WBC count time has 4 seconds differences up software all is done. I hope I may get solution from you for my problem faced?
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I think if you reduce the length of the suction tube by 4 seconds. Your problem will be solved.
Companies normally give a tube for extra length, and this makes the sample to go into cuvette after a long time. So send the sample quick the tube length should be optimised. The cut additional length can be after washing kept in stock.
I suspect this might resolve the issue. And it's nothing to do with software. We had a similar issue on a different equipment.
Ignore the earlier answer that answer was based on the way the instrument was finctioning. Sorry for the delayed response.
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A strong Association has been described between Blood group O and Peptic ulcer, Blood group AB and Carcinoma cervix, Blood group A and Gastric Carcinoma , Blood group B and Pemphigus and Seborrhoric Dermatitis.
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Here is what the lab said when I sent out my samples to have CBC ran, the first time I sent out they said samples were clotted. I thought it must be due to not inverting sample EDTA microtainer enough. Also I switched from saphenous vein to the tail vein for the second collection. All samples were taken beautifully and I had at least 400ul of whoole blood in each tube. The samples sat at room temperature then packaged in a cooler with fridge packs not touching samples in box. Next day here are my results:
Hematology samples received this morning. Some samples were grainy. See attached picture. We made slides on for them but some just look unusable. Some of the slides did not contain a mono layer to where we can make an appropriate differential (distinguish between the different types of white blood cells). RBC’s and WBC’s were disintegrated and hard to tell what they are.
We tried running a few of the samples and the values does not look valid either. The hgb (hemoglobin) is the same value as the hematocrit.
Can ANYBODY help me?? Explain why this would happen, mu boss isn't very happy right now...
ANy help would be so much appreciated.
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One cause could be the cooling for which I propose 2 solutions (the first is the best):
1. Take the smear immediately after collection
2. return the tubes to room temperature after they have been removed from the refrigerator
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Can anyone share a protocol for a ferrozine assay for tissue samples that is reproducible, including the lysis protocol? Also, what is the minimal amount of iron detectable using this protocol and which method was used to quantify the tissue amount (by protein?)?
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Respected sir,
i want to estimate non heme iron in Fe+2 state, in a protein. Please tell what method i can use? It is very urgent
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We are reaching you out because we are working on a project at the RWTH university Aachen, called Epi-Blood, which enables counting of leukocytes in frozen blood retrospectively, with high accuracy and requires only low volumes. www.Epi-Blood.de
We would like to understand the demand of the service that we will provide in the research sector. Therefore, we ask for just 2 minutes of your time to answer only 5 multiple choice questions. If you feel like contributing, please use the link below to start the survey. Thank you for your participation!
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I submitted a survey. Best of luck!
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I work in a cancer lab and have been trying to isolate and quantify platelets without the use of any advanced hematological instruments. I don't need pure platelets, so I've just been generating platelet-rich plasma and trying to get a rough count using a neubauer chamber. With this method though it's incredibly difficult to figure out what I'm looking at under the microscope, and counting platelets has been impossible. Does anyone have experience with this?
Current PRP isolation protocol:
1. Collect blood from mice after CO2 euthanasia via the posterior vena cava, collect into sodium citrate (1:9)
2. Centrifuge whole blood @300g for 20min
3. Collect upper 2/3 plasma fraction
4. Centrifuge fraction @800g for 15min
5. Resuspend in PIPES buffer, pH 7.4
6. Dilute 1:20 and add 10uL to Neubauer chamber
7. Allow cells to settle in moist environment for 15min, try to count
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Dear Ma'am Savannah Free please read these articles, they might be helpful for your study.
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Hi,
I have a question regarding the hematology analyzer for complete blood count for blood samples. I would need to analyze fixed blood samples and I would like to know if the analyzer could work also with this kind of samples. I know that the analyzer detects electric properties of the cells and with fixed samples I guess it won't be possible. Does anybody have experience with that? Thank you for your help!
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Hi Mara
You may want to have a look at this paper. Good luck
Siva
doi: 10.1016/j.jim.2010.09.029. Epub 2010 Sep 27.. 2011 Jan 5;363(2):158-65.J Immunol Methods
Stability of immunophenotypic markers in fixed peripheral blood for extended analysis using flow cytometry
Cuc Davis 1, Xiaoying Wu, Weimin Li, Hongtao Fan, Manjula Reddy
Affiliations expand
  • PMID: 20875824
  • DOI: 10.1016/j.jim.2010.09.029
Abstract
The analysis of whole blood samples by flow cytometry for pharmacodynamic and biomarker assessments in clinical studies has been limited by the necessity to test these samples within a short time frame after blood collection. In most clinical studies, blood specimens are shipped to a centralized testing facility; it is critical to demonstrate specimen stability over a period of time which will encompass the time elapsed between specimen collection and testing. A possible solution to overcome this limitation is the use of a fixative to preserve the cell surface antigen stability in whole blood. We examined the stability of markers for T lymphocytes (CD3, CD4, CD8, CD45RA, and CD45RO), B lymphocytes (CD19), NK cells (CD16+CD56), activation (CD25 and HLA-DR), chemokine receptors (CCR5 and CXCR3) and skin homing (CLA) in fixed blood over 7 days and used this information to select the markers for global clinical studies. These assays with selected markers were further validated using fit-for-purpose approach (Lee et al., 2006) and to set the sample acceptability criteria for use in clinical sample testing. Most of the markers examined were stable when collected in Cyto-Chex® BCT for one week with the exception of the activation markers on T cells
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Higher affinity for CO induce suffocation which may be fatal.
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In agreement with Pranita Kamble Waghmare, the Oxygen axis after oxygen binding with heme is at an angle while Carbon monoxide binds to free heme with the CO axis perpendicular to the plane of the porphyrin ring via carbon-Iron bonding. So, the two oxygen atoms in oxygen exhibit steric hindrances on each other. In which case, CO doesn't experience the same.
This perpendicular orientation is favorable for Hb binding.
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The website addressed to the above link goes inaccessible at times. The reasons are unknown. This is a key database accessed by many hematopoiesis researchers. if anyone knows alternative ways to access the database, kindly post here, which will be useful for many researchers. Thank you.
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I did not test, but I think the page is working
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The use of the RefVal freeware for establishment of blood reference intervals for wildlife species has been popular in recent years, however the add-in doesn't appear to be compatible with Excel in Office 365. Has anyone else got around this problem, or if not what are you using that's equivalent? MedCalc is good, but expensive. Thanks!
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I am currently doing an experiment which involves changing the techniques in staining of blood smear. I use expired EDTA tubes to store the blood collected, does the expired tube will affect the morphology of blood in any way when I view it under the light microscope? 
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If a blood collection tube is used past its expiration date, the vacuum may not draw the amount of blood needed to fill the tube completely. ... If the tube contains an anticoagulant, it may not work effectively (may not prevent the blood from clotting)
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There has been evidence that those with minor imbalance of blood cells [for unspecified condition], in which the red blood cells are small and less in quantity than white blood cells, can be of benefit by a dosage of Vitamin C and daily moderate exercise.
When in the case of shortness of breath, while at the same time having such small in size red blood cells, were able to show benefit by a dosage or iron and Vitamin C in a single dose. (this was not a contraindication of the particular case)
Can you verify that cases of unspecified shortness of breath that may be correlated to small-sized red blood cells, can benefit from a monitored vitamin dosage?
Similarly, Co-Q10 shows benefit for those who may have heart damage. Of course, a doctor must manage patient supplements.
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Studies show that consuming more vitamin C can increase your blood antioxidant levels by up to 30%. This helps the body's natural defenses fight inflammation ( 4 , 5). Vitamin C is a strong antioxidant that can boost your blood antioxidant levels. This may help reduce the risk of chronic diseases like heart disease.
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We have recently seen a case of large diffuse B-cell lymphoma of probable folicular origin (CD10+ partial expressión) with a well defined CD103 expression by B-cells. The references in the literature are scanty and unclear.
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Jordi Juncà We have just analyzed the blood of a patient with previous diagnosis of follicular lymphoma in lymph node that returned to the hospital with adenopatias and abnormal lymphocytes in blood. We performed an immunophenotyping of the blood and found 12% of medium sized cells with characteristics of follicular lymphoma: CD19 weak, CD10++, BCL-2++, CD5 neg, CD43 neg and IgM neg. However, these cells showed some positivity for post-germinal markers such as CD27, CD11c and CD39. In addition, 50% of the abnormal population showed expression of CD103 (without expression of CD25, CD123 or LAIR-1). It is the first time I see CD103 expression in a follicular lymphoma. On the other hand, I´ve already seen cases of hairy cell leukemia (very classic) with CD10 expression.
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I am trying to compare fibrinolysis between two genotypes of mice after an insult. The D-Dimer levels were equal between the mice but I was told that rapid fibrinolysis (which we're expecting) would form fibrin degradation products instead of D-dimers.
I've found a few ELISAs of unknown quality but I'd really like an antibody I can use for western blot. Preferably I'd like one that can detect multiple fragments so I can see if production of one fragment is different over the others. One of the main issues so far is that there are very few hematology reagents for mice. Most are for humans.
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I think that you should focus on an assay that quantifies degraded fibrinogen rather than degraded fibrin (which is typically consists in D-dimer and some fragmented fibrin)
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i need this information for preparing fish feed to analyze the effects of nano particles on fish growth and hematology.
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Dear Farkhanda Asad just in case that this is still of interest to you, please have a look at the following relevant article:
Effects of dietary iron oxide nanoparticles on the growth performance, biochemical constituents and physiological stress responses of the giant freshwater prawnMacrobrachium rosenbergii post-larvae
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We are currently using Hemavet and Vetscan but they are both very old and we are looking to upgrade. Thanks!
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Thank you both!
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I am doing a master in nursing but still does not have any idea about what topic I am going to choose. pls help
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Dear Siti
Potentially there are numerous topics you can go for but the current Corvid 19 pandemic does offer Haematologists opportunities to investigate various clinical aspects, manifestations and complications of this condition. You may want to choose a disease category first (BMT, acute leukaemia, chronic leukaemia, MPD, MDS, haemophilia, haemoglobinopathy, non-malignant cytopenia....). Then analyse the impact that Corvid 19 and lockdown have had on the presentations and management pathways of one of these categories. You can analyse the patient numbers (up or down), any difference in the way patients present to hospital (eg. more severe disease, significant complications), any unusual presentations, any notable differences in laboratory parameters (more marked cytopenia, biochemical disturbances etc), differences in clinical outcome (morbidity and mortality).
I think that should keep you busy.
Good luck with your thesis.
Siva
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Regards
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The focus and scope of this journal are related to various areas in the field of medical laboratory technology such as in the field of parasitology, bacteriology, hematology, clinical chemistry and other areas related to the field of medical laboratory technology.
There is no charge for the entire publishing process in this journal.
In addition, we also invite you to become an editor or reviewer in this journal.
For further information, you can visit our journal at www.teknolabjournal.com or via email at budi.setiawan@poltekkesjogja.ac.id
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I am Assistant Professor of Hematology and Immunology, Faculty of Medical Laboratory Sciences, University of Gezira, Sudan
I am intertesting in publishing my scientific papers in this Journal.
In addition, also I am intertesting to invite me to become an editor or/and reviewer in this journal.
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I was posed a question by a hematopathologist in training this morning: Why are B-cell lymphomas more prevalent than T-cell Lymphomas?
My initial guess was perhaps there is increased tolerance training in T-cell populations, but it turned out she did not know either. Her best guess was that it is multifactorial in nature, i.e. genetics, viral infections (HTLV), and environmental factors.
Does anyone have any papers or book chapters for reference? I would love to be able to bring a more detailed and concise answer back to her. So far, I haven't found anything that answers the question on PubMed.
Thanks in advance.
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Development, activation and contraction is more tightly regulated in T cells v B cells. Particularly the latter, so you are less likely to see t cell lymphoma secondary to chronic immune stimulation for example malt lymphoma. Also, ebv. B cells express cd21 whereas t cells only transiently do so during thymus development. So ebv causes hl, bl, mzl, malt, and dlcbl in b cells but basically, only extranodal nk/t cell lypmphomas in the cells as I recall. 97% of adults are ebv seropositive.dlbcl is the most common lymphoma in the us, roughly 15% is eber+.
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Hello, I am an undergraduate student . Currently I'm working on "Hematological diagnosis using blood sample images " project . I am working on developing a deep learning model for this diagnosis . Now I need useful research materials to continue my research . It will be really helpful if free research papers were provided regarding this topic.
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Two books authored by Prof. Barbara Bain may help you.
1. Practical hematology
2. Bone marrow pathology
Also you may read my research on hematological diseases including anemias and leukemias, etc. You can find them at my profile
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I'm trying to set up a protocol for PBMC isolation from whole blood using density gradient centrifugation. In a previous lab we diluted the blood in RPMI prior to layering, and washed the PBMCs in RPMI, but a lot of protocols I've seen use PBS instead. Is there any reason to use one or the other, or do they both perform the same function?
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Using RPMI is recommended if the cells are to be cultured and check for viability since RPMI will enhance PBMC yield
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Haemoglobin is a serious problem during purification and analysis of antioxidant enzymes from red blood cells. For this reason it could be necessary to remove Hb in order to obtain a clear supernatant.
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Thank you for the answers on how to remove haemoglobin in erythrocytes lysates. the answers have brightened my day.
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It's been a while, while some patients in whom high grade fever, prolonged duration of fever doesn't subside and no early diagnostic tests and hematological examination turn out to be positive, fungal infection has been seen either in urine or even in blood culture in few patients. Non albican type fungal infection seems to be one of the important cause of high grade fever in spinal injured patients.
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follow
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Hi. Good day. I am currently working on ROC Curves to determine the *specificity, specificity, negative & positive predictives value and the likelihood ratio of pos & negative values. I am looking for cut off points in IDA of its hematological markers (FBC, FBP, RDW, MCH and MCV). I would like to ask, if i manage to plot the curve using SPSS, it will automatically provide me with *these values or do i need to do crosstabs or other test to find these. Manually there is a formula to calculate these, but in spss? and how to discuss the output. Thanks
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The sensitivity, specificity etc vary according to the location of the cutoff in the range of the biomarker/predictor. The selection of the cutoff depends on the cost of false positives and negatives. There are automatic methods of cutoff selection such as maximising the Youden criterion (that maximises sensitivity+specificity). It is also important to realise that it is that possible the actual predictions from the biomarker may be inaccurate even if there is good discrimination based on the ROC cufve (AUROC, sensitivity an specificity). Accuracy of the individual predictions from a logistic regression can be assessed using calibration techniques. Additionally all model assessment is best carried out using cross validation of in test/training splits. There is more info here https://www.youtube.com/watch?v=v_e85TDYj9Y
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Biological techniques are methods or procedures that are used to study living things. They include experimental and computational methods, approaches, protocols and tools for biological research.
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Is bcl2+ in DLBCL always associated with poor outcome in PFS or OS, or is this finding significant only in high IPI score DLBCL?
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Nowadays, where there is a lot of new information and studies on immunotherapy in Hematology and Oncology every day, are there news about Haemophagocytosis and Immuntherapy, such as Antibody against PD-L1 AS Exemple?.
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More than 2 Years sinnce my Quastion about it in 2018 😀 ...
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Imatinib is a tyrosine kinase inhibitor and used as cancer drug for some hematological cancers.
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Imaginaba is a specific inhibitor at the bcr-able chromosomic rearrengement. Why do you think it will inhibit other kinases like cell surface growth factor receptors?
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Any synthetic analogue of Hemoglobin or any other Xenobiological agents can be cited as example. Please provide links and references.
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Myo-inositol trispyrophospate (ITPP) shifts the oxyhemoglobin dissociation curve to the right, thus decreases hemoglobin affinity for oxygen and results in an increased oxygen delivery to the tissue. I has been considerd as a potential agent used in dopping of athlete and a performance-enhancing substance in horse racing.
Biolo, A; Greferath, R; Siwik, DA; Qin, F; Valsky, E; Fylaktakidou, KC; Pothukanuri, S; Duarte, CD; Schwarz, RP; Lehn, JM; Nicolau, C; Colucci, WS (2009). "Enhanced exercise capacity in mice with severe heart failure treated with an allosteric effector of hemoglobin, myo-inositol trispyrophosphate". Proc Natl Acad Sci U S A. 106 (6): 1926–1929.
Lam, G; Zhao, S; Sandhu, J; Yi, R; Loganathan, D; Morrissey, B (2014). "Detection of myo-inositol tris pyrophosphate (ITPP) in equine following an administration of ITPP". Drug Test. Anal. 6 (3): 268–276
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Hey there,
I'm using commercially available Drabkin's reagent to measure the hemoglobin content of blood photometrically. The reagent I bought contains sodium bicarbonate, potassium ferricyanide and potassium cyanide. In my understanding, all different forms of hemoglobin (except sulfhemoglobin) are converted to methhemoglobin by potassium ferricyanide in a first step which reacts then with potassium cyanide to cyanomethemoglobin which shows a significant absorbance at 540 nm.
But sometimes I see a second peak around 575 nm (indicated by the red arrow in the graphic), sometimes very weak but sometimes also very strong and I wonder what it could be. Could this be some remainings due to uncomplete reaction or could it maybe be due to poor quality of the sample?
Thank you for your advice!
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Oxy-Hemoglobin typically has peaks at 541 and 576 nm. Because these peaks are still present in the spectra you showed, your sample likely still has some amount of unreacted hemoglobin which will impact your results. Typically this lack of complete reaction is caused by 2 factors:
  1. Insufficient Drabkins reagent
  2. Not enough time for the reaction to reach completion.
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How would you score Daflon (Diosmin and Hesperidin) with regard to blood thinning effect? How would you rate it in comparison with Aspirin, Warfarin, other agents? I.E do you consider it to be potent? Mild, moderate?
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Hesperidin might prolong clotting time and worsen underlying bleeding disorders
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Does anybody know how many CD34 marker exist on the surface of each CD34+ hematopoietic stem/progenitor cell?
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Я згодна з Hasan lssa та Салех Алкарім . Бажаю удачі в наукових дослідженнях
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I stained mouse peripheral blood with B220 and CD3 markers and found all the CD3 postivie cells were B220 positive too. My aim was to distinguish simply B cell and T cell by these markers but I am confused now. Any comments, or advice?
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Autoimmune phenotype, as in autoimmunity-prone Fas-mutant lpr/lpr mice.
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I plan to conduct a study in which I will compare depression and quality of life among children with cancer to children with other chronic conditions (hematologic, kidney and respiratory chronic conditions). Is it appropriate to say that the cancer patients are case and other children are control. Or should I change the study design?
In case my case group are children with cancer, who should be the control group?
Thanks
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If your research question is "does cancer cause depression and loss of life quality" you should compare non cancer patients with cancer patients ie start with the "exposure = cancer" and compare the exposed group to a non exposed group otherwise similar, This is a cohort design, not a case-control design.
if on the other hand you have a group of patients with depression and poor life quality (the cases) you should select a group without depression or poor life quality (the controls) and compare the presence or absence of cancer in the in the two groups. This would be a case control study..
Ii would recommend the cohort approach.
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Do you consider this be thrombocytopenia?
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Thrombocytopenias with the presence of normal functioning thrombocytes may provide some survival advantages for human being since myocardial infarction and stroke are the most common causes of death in developed countries at the moment, and the roles of thrombocytes in such events are significant. Thus initiation of low dose aspirin is strongly advised above the age of 50 years.
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I need to differentiate HSCs towards granulocytic lineage, though I have no access to IL-3. I was wondering if I can do the above only using GCSF, FLT3, and SCF.
I appreciate your consideration in advance.
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Yes by sorting the cells.
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Dear Colleagues,
My Dad (66 years old) has recently beed recently diagnosed with multiple myeloma. Apart from significantly elevated monoclonal protein, he has no symptoms. He has had his blood protein levels high for the past 5 years. Detailed medical exams showed that his bones, kidneys, and blood work (other than what I have already mentioned) are all fine. My Dad has always been fit and he is still feeling well overall.
He has an opportunity to join a clinical study that uses daratumumab combined with standard treatment for multiple myeloma. My understanding is that the treatment he would get is considered chemotherapy.
Is undergoing the chemotherapy the right thing to do now, considering multiple side effects of it (with or without daratumumab)? Or should he wait until symptoms begin to appear. My Dad’s doctors do not seem to share the same opinion on this matter.
I would deeply appreciate suggestions. Please note that I am a neurolignuist, not an MD. Therefore, I may have worded something incorrectly here. However, I will gladly clarify or provide more detailed information if helpful.
Thank you very much in advance,
Monika
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Dear Monika,
The exact diagnosis of your father is probably smoldering multiple myeloma (SMM), according to the information you gave about his medical data. SMM is defined as following criteria:
Serum monoclonal protein: IgG or IgA ≥3 g/dL, or
Bence Jones protein ≥500 mg/24 h  and/or
Clonal bone marrow plasma cells between 10% and 60% and
Absence of myeloma-defining events or amyloidosis.
The risk of progression to symptomatic multiple myeloma of SMM is about 10% annually.
Experts in hematology consider SMM as an excellent setting to test the efficacy of novel theurepatic drugs such as ixazomib, pomalidomide, elotuzumab, and daratumumab in myeloma treatment.
Despite standard care of SMM is close clinical observation until development of symptoms, International Myeloma Working Group recommends therapy for SMM cases at high risk (about 50% risk for progression to symptomatic multiple myeloma).
For more information please visit:
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I need research and literatures for my MSc Thesis, effect of petroleum refinery effluent on albino rats using moringa plant or seed extract as abatement. There are few research and study conducted so far on this that i have found. Could be that the effluent are completely free from pollutants which can be toxicant to human. Could we suggest that the industrial waste water are completely processed and treated before discharge to the environment?
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please provide more literature review especially its effect on hematology and histopathology
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Hematology statistical analysis of HB and Blood Group Vs Age and Weight.
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GraphPad Prism is very easy to use and teach very well what test you have to do
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This is a challenging case I've been facing, about a 65-year-old lady presenting with weekly recurrences of herpes infection on sacrum and inner thigh (most likely HSV type 2) in the last 20 years. Recurrent vaginal candidiasis coexists. I suspected of acquired immunosuppression and her immunological panel revealed:
leukocytes 1,908
lymphocytes 816
CD4+ 542
CD8+ 166
IgA 44mg/dl
Hb 14,4g%
plat 219,000
Complement levels and other immunoglobulins are within the normal range.
She takes the following drugs continuously (could any of them explain leukopenia + lymphopenia?): omeprazole, ranitidine, amitriptyline, desvenlafaxine, pregabalin, and vaginal ovules of boric acid.
I prescribed imiquimod cream locally, 3 times a week, and she reports parcial improvement of frequency and severity of skin infection.
My doubts now are regarding the continuation of propaedeutics and therapeutics. Any thoughts? Relevant literature?
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i think you should continuous in treatment of local lesion with stop the therapeutic after doing the level of lymphocytes and WBC count
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I am studying about reference interval of hematology parameter followed CLSI guideline. Then i got some difficulties in percentage of eosinophil. I got my study subject have >0 value. But in Robust method i got minus lower limit? How it be possible? Anyone can help me to raise my lower limit?
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Yes, with very low values you can get a negative reference interval. I believe the standard protocol would be to simply put 0-your max (as clearly a negative result would never be encountered in the field). Eosinophils (% of WBC) are usually 0-1 ish in the species I work with.
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If we get spherocytes on peripheral smear of blood, will it be enough to diagnose Hereditary spherocytosis
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I agree with Christos answer regarding the role of both Ankrin and Spectrin deficiency in the RBC of the HS. The description of different immunoassays methods is also important. However, I think that in answering a question in RG it should be considered the country of origin, wondering if technically sophisticated diagnostic methods are available in those places. I often stick to this rule, limiting myself to suggest only diagnostic strategies hypothetically (or realistically) available to the Colleague who poses the question
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Hello, could anyone share your experience regarding CBC analysis in sinovial fluid samples and pretreatment with hyaluronidase? Is it absolutely necessary for every sample to be treated with the enzyme? Our equipment include Siemens Advia 2120i hematology analyzer.
Thank you!
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Not much sure but one risk which i had been anticipating are:
1- The very high viscosity of the synovial fluid, which cause the automated cell counter to get clogged so pre & post run heavy detergent wash seem to be appropriate.
2- Secondly, we have exact internal and external QC samples for normal blood counts but usually markets in our set up do not provide specific QC samples for other fluids. So whatever counter data with internal and external QC validation is not reliable. If any body knows specific internal QC sample, kindly let me know as well.
3- Thirdly i still sometime run them sometimes directly and some times with 1:1 dilution (to help avoid clogging of my counter) BUT my main reliance is on manual inspection of those cells by microscopy.
4- Antonio La Gioia highlighted the idea of pre-treatment with hyaluronidase, which seem promising but i have no experience of working with that. Here, i would request if he had the data, kindly attach it as a link.
5- Finally, i guess it is also important what we actually need to see within the specimen. Mostly there are polymorphs so what is pre-test odds one is targeting while seeing a synovial fluid?
Regards
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i m working on hematology of silver carp(Hypophthalmichthys molitrix).i want to know the value of hemoglobin, RBC count, WBC count MCV, PCV, MCHC, neutrophils, eosinophils, lymphocytes and monocyted of a healthy silver carp.kindly guide me?
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Hello
There is rarely blood reference intervals in the fishes, including silver carp. Although there are some paper with title of "Blood reference interval of fish ...", you should note that the fish blood characteristics are very variable depending on environmental condition, fish age, nutritional history, sampling protocol and fish strain. This is the reason for assigning "control group" in any studies on fish.
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Dear all friends,
I am a graduated PhD student of laboratory hematology and blood bank from Iran. I am so keen to continue my studies in Postdoc grade particularly in Germany. I have strong potentials to do research activities as work-team. My priorities are cellular biology and stem cell-related experiments. If anyone knows institutions that I can apply for a Postdoc position, please notify me. Also, if there is any necessity to send my CV for more information, please do not hesitate to tell me. Thanks in advance for your entrancement.
Kind Wishes
Alireza Goodarzi
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Hi Emmanuel,
Thank you for your kind response:)
Best
Alireza
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Hematological parameters
I will so much appreciate your response
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Thank you all for your helping responses..
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I want to make a stable blood sample. In my study, I want to make a comparative study of many hematology devices and I want to try many blood sample from different sources.
I've seen many protocols, all of them old Patents, I don't know which I should depend on.
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At our Institution we have a protocol (WHO/LAB 97.2) for whole blood preservation (glutaraldehide 0,37%, formaldehide 2,7%, trisodic citrate 26%, and antibiotics) which is intended, precisely, for CBC blood counters. You can seek it in the WHO page. Success!
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Minimum of 24 hours and maximum of 48 hours. But it depends on your area of interest
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Every hematology textbook, the chapter of idendtification of common Hb variants mention about electrohoresis in agar gel with citrate buffer at pH 6.
I have discussed with many eminent hematologists in India and found that nobody is using this method any more because of new technology available, but also none of them have first hand experience about the method. Why so ?
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Try doing it once and write here what are your experiences
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I need two detailed and reliable lab protocols one for assessing NK cytotoxicity and one for NK degranulation assay using flow cytometry.
Is NK cytotoxicity done only in co-culture with K562 or any other hematological cell line such as U937 can be used instead?
I would highly appreciate if anyone could help.
Thank you in advance!
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Both cytotoxicity and degranulation assays can easily be performed using a four-color flow cytometer. For cytotoxicity, target cells can be pre-stained with CFSE. After incubation with NK cells at different E/T ratios, cells are stained with a live/dead marker. You can use traditional dyes like PI or 7-AAD, but we normally use LIVE/DEAD™ Fixable Far Red Dead Cell Stain Kit from Invitrogen in the “APC” (aka FL-4) channel.
In theory, degranulation and cytotoxicity can be determined in the same well using a PE-labeled antibody to CD107. However, cytotoxicity is more effective at higher E/T ratios, while degranulation responses are stronger at low E/T ratios.
Natural cytotoxicity can be assessed using the cell lines indicated above – K562 cells are readily killed by resting NK cells (i.e. not cytokine-stimulated). For other cell lines, pre-stimulation by IL-2, IL-12 or IL-15 may be needed to observe robust cytotoxicity responses.
By combining anti-CD20 antibodies (e.g. Rituximab) and CD20-expressing target cells (e.g. 721.221 or Daudi cells) you can also study NK cell-mediated ADCC.
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This is the sevent exercise regarding my proposal to share our professional experiences in hematologic morphology. Most of previous exercises aroused interest and participation. Some, such as the sixth, have not piqued your interest. I propose the seventh case that I hope will generate interest and participation.
There are images from BM aspirate of a 79-year-old man with pan-cytopenia. Numerous cells have blastic features: increased size, exiguous cytoplasm, dispersed nuclear chromatin without evidence of nucleoli. Immature granulocytes and NRBC are present.
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I wanted to share on RG this interesting case because it allows us to discuss some difficulties in morphological interpretation that can lead to relevant diagnostic errors. "Blast cells" which I described having increased size, exiguous cytoplasm, dispersed nuclear chromatin without evidence of nucleoli, might be classified as lymphoblasts. So a diagnosis of ALL could made. However, a careful observation of the images allows to detect some cellular fusions (Figure A on the left; Figure C right).The figure C, D and E below show the ability of these cells to form syncytia. Actually this is not an acute leukemia but a metastatic lung cancer (small) in the bone marrow
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Hi there! I am wondering if there is any way of transiently increase platelet count in mice, like administrating TPO or something like that?. Thanks a lot!
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Thanks everyone for the nice comments, finnaly I have decided to use a TPO mimetic drug that seems to work in mice as well. Nevertheless, good to know about HST1 as well. Best.
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Who suggest these pictures?
In the attached photos we can observe 2 microscopic fields from a PB smear of a 27-year-old woman that went in the emergency room with a serious breakthrough bleeding. In addition, she had many bruising in many parts of the body. CBC showed HGB 127g/L, WBC 23x109/L and PLT21x109/L. The doctor on duty at night was not a laboratory hematologist, so he asked for help from a skilled collegue that after seeing the slide alerted the emergency room of the seriousness of the case.
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This is typical APL case with hypogranular promylocytes. There are distinct morphologic features such as bilobed nucleus and paucity or absence of granules. These cells will be classified as blast equivalent. A lot of times these cells are confused with monoblasts. In this case, presence of Auer Rod would help to differentiate one from another. Some cases, however, do not have obvious cytoplasmic granules, may have basophilic cytoplasm, and have folded bilobed nuclei that may be mistaken for monocytes. Of course, diagnosis won't be made until cytogenetic and flow cytometry are done. However, laboratory hematologist should be able to recognize the morphology to alert physician a possibility of DIC.
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First, I'm using Ficoll Histopaque centrifugation to obtain mononuclear cells. Then, I'm doing RBC lysis. I observe a drastic loss of CD138+ cells after that (as checked using FACS and two separate validated CD138 antibodies). E.g. a sample that contains 18% plasmocytes, has practically no plasmocytes left after Histopaque isolation.
I want to use MACS CD138+ Microbeads but they fail since the Histpaque-isolated cells are devoid of plasmocytes.
Does anyone have such problems? What would you recommend? I've read about autoMACS Pro being good for such cases, but I don't have access to this device.
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We do at first Ficoll to obtain the MNCs but without Erythrocte lysis and then go for CD138+ beads for purification from Miltenyi. We do this manually with a magnet from Miltenyi and the respective column which is working as good as automated system, only requires some person to do it. The number of CD138+ cells strongly depends from the patient, often we get either nothing (everything below 50.000 cells is considered in our lab as nothing), or in between 100.000 and 200.000 cells from 20 ml of bone marrow. In rare cases we obtain more than 1 mio cells (up to approx. 10 mio cells was the maximum I got), this is strongly associated with disease progress. Try to get patients history and cases with disease progress for method establishment as this can only be successful when you have a proper amount of cells inside. And also do not hesitate to contact Miltenyi that they should come and help you, my experience in the past was that they did this and also gave free kit samples for testing purpose or borrowed even a magnet for 4 weeks for testing.
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Since automated hematology analyzers use staining with nucleic acid specific dyes to detect immature platelets I am curious to learn whether these dyes would also detect nucleic acids from pathogens associated with platelets. I know of a case where Babesiosis was originally overlooked due to a high reticulocyte count and assume that what can happen for red blood cells could potentially happen for platelets as well. Differentiation between immature platelets and "pathogen-associated" platelets might be of particular importance in diseases like autoimmune thrombocytopenic purpura. It could help our understanding of platelets as innate immune cells if immature platelets could also represent "pathogen-associated" platelets.
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Dear Elizabeth
Very interesting and thought-provoking question indeed. Once upon a time, I did some experiments using thiazole orange that stains mRNA to differentiate between essential thrombocythaemia and reactive thrombocytosis (RT) on the basis that one would expect to see more mRNA in RT. The results did support this assumption. Of course, lots of technological advances have happened since. But, I think, one could use nucleic acid stains to differentiate between reticulated platelets and pathogen-infected ones. It will be very interesting to study. Since we have more advanced dyes for flow cytometry, you could run some experiments with a combination of RNA specific and DNA specific dyes using platelets from known cases of RT. You may find that RT platelets express more RNA rather than DNA. Once you have established the expression profile, you can then study platelets from patients with confirmed bacterial septicaemia, viraemia (HIV, CMV (allo-BMT), acute glandular fever...) and parasitaemia (malaria, babesiosis...) to look at any difference(s) in the profile. The results may give you some useful clues for further research. Good luck with the experiments and please keep me in the loop. I would be very interested to know what you find.
Best wishes
Siva
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I would specifically like to culture Type 3 Innate Lymphoid Cells
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Did you succeed to culture innate lymphoid cells in vitro ?
How did you do it and what culture medium did you use?
Thanks for you answer
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Any hematological slides for selling?
You can send by post
Contact please
we need parasitological finds such as Tripanossoma, Babesian, Bacteria infection
Cell inclusion: basofilic stip, .....
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I think you can easily prepare them by your self
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I was asked to supervise my junior in a small project about Chick hematology, because I am new to the topic I have several questions to ask :
1. My juniors are planning to see the garlic oil effect on the chick's white blood cell counts, is it possible to apply oral gavage on chick as the enteral administration method?
2. Is there any suggestion on substance that affect overall blood cell count that doesn't need oral gavage administration like syringe usage ? we are prohibited from using heavy metal
3. If you have any reference about chick hematology-related research please let me know
Thank you
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Dear Grace,
I think the following recent papers will help you:
Owosibo AO, Odetola OM, Odunsi OO, et al. Growth, haematology and serum biochemistry of broilers fed probiotics based diets. African Journal of Agricultural Research 2013; 8(41):5076-5081. http://www.academicjournals.org/journal/AJAR/article-full-text-pdf/3A5206141348
Akinola LAF, Etuk MO. .Haematological and Serum Biochemical Responses of Broilers Fed Varying Levels of Indomie Waste-Based Diets. IOSR Journal of Agriculture and Veterinary Science 2015;8(3):66-70. https://pdfs.semanticscholar.org/995e/f4f36bb59e9f1db0e48ae27a924a2cb8dda8.pdf
Abdulazeez H, Adamu SB, Igwebuike JU, Gwayo GJ, Muhammad AI. Haematology and Serum Biochemistry of Broiler Chickens Fed Graded Levels of Baobab (Adansonia digitata L.) Seed Meal. IOSR Journal of Agriculture and Veterinary Science 2016; 9(10):48-53. http://www.iosrjournals.org/iosr-javs/papers/Vol9-Issue10/Version-2/I0910024853.pdf
Ogunwole OA, Abu OA, Adedeji BS, et al. Haematology and Serum Indices of Finisher Broiler Chickens Fed Acidified Blood Meal-based Diets. Journal of Advances in Biology & Biotechnology 2017;11(2):1-7. https://www.researchgate.net/profile/Olugbenga_Ogunwole4/publication/312476729_Haematology_and_Serum_Indices_of_Finisher_Broiler_Chickens_Fed_Acidified_Blood_Meal-based_Diets/links/587f12e908aed3826af46bb9.pdf
Best wishes from Germany,
Martin
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I have a silly question about the OR. I conducted a study about adherence on oral oncolytics and found that the presence of a solid tumor (OR = 2,026; 95% CI 1,058-3,880; p=0,033) and cyclic dosage regimen (OR 9,457; 95% CI 3,909-22,877; p=0,000) were significantly associated with optimal adherence.
So does this also imply that a hematological tumor and a continuous dosage regimen are significantly associated with non-adherence?? If so, than the ORs for these variables are 1/OR?
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Thank you Marcelo and Olena for helping me out.
Now i am confused how to interpret an odds ratio<1.
I have found that a continuous dosing regimen (OR: 0,106; 95% CI 0,044-0,256; p=0,000) and a hematological tumor (OR: 0,494; 95% CI 0,258-0,946; p=0,033) are significantly associated with non-adherence. But how should i interpret the OR's? Patients with a continuous dosing regimen have a chance of 89,4% to be non-adherent?
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Bone marrow aspiration of a 84-yer-old man with anemia (HGB 100g/L), leukocytosi (75x109/L) and thrombocytopenia (47x109/L). Blast cells in PB 37%. In the attached photos some microscopic field of PB (1) and of bone marrow (2-3).
I would like to discuss about this case with RG community
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The morpholgical features of monocytes had been described in the WHO 2008 classification to differentiate monoblasts from pro-monocytes and pro-monocytes from abnormal monocytes (figure 1). Pro-monocytes are considered "monoblasts equivalents" when the percentage of blasts is required for the diagnose of different types of acute leukemias of monocyte lineage. It's important that abnormal monocytes are excluded from the blasts count.
Previous photos are from an acute monoblastic leukemia (acute myeloid leukemia not otherwise specified in the WHO classification; M5b in the FAB classification) and constitute an interesting exercise to distinguish different types of monocytes. In the figure 2, a naphtyl butyrate esterase staining of bone marrow.
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Person who has Vitiligo is permanent deferral for donation or can donate blood for the use of blood transfusion in needy patients .
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Thanks Jyoti
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In the attached photos we can see some cells having similar morphology that in a 65 old years man with middle anaemia (90 g/L) were 23% of a total 5200 lymphocytes. What suggest these?
Because I know the final diagnosis, this is a further contribution to the discussion  and sharing of professional experiences in haematological morphology.
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Hi Antonio,
Prolymphocytic leukaemia (PLL) is my first differential. Meanwhile, does the patient has organomegaly, especially splenomegaly?
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Please share me the research if you come across on exogenous enzymes supplementation to fish feed and fish haematological status.
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Dear Sir, none of your suggested publications are related to my query. Anyway thanks for your effort. 
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Should clinical haematologists put their microscopes on E-Bay? (van't Veer M, Haferlach T. Haematologica, 2014). 
I think no. The ability in recognizing cellular morphology in  peripheral blood and in the bone marrow smears is a crucial step in the diagnostic process of haematological diseases even today. Unfortunately we're losing this expertise also because we are unable to transfer our experiences to new generations of professionals. ResearchGate is a formidable place of sharing and exchanging experiences. So, I invite the morphologists to participate in a kind of forum like "images on haematology" with exchange of images, comments and clinical information regarding haematological cases of interest.
I start with images of extreme dysgranulopoiesis in a case of AREB-1
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Haemophagocytic lymphohistiocytosis is a hyper-inflammatory syndrome which can occurs in a primary familiar (fHLH) form in the paediatric age and in a secondary form prevalently in adult age (sHLH). Both fHLH and sHLH are characterized by a severe clinical onset with fever, hepato-splenomegaly, bi-, three-cytopenia in PB, hemophagocytosis in the BM as well as BM and multi-organ failure. sHLH is more commonly associated with other pathological condition including malignancies, immunodeficiency, infections, rheumatic diseases and autoimmune diseases. In the majority of cases a “trigger” event such as infections (by Epstein-Barr virus in primis) or drugs (anti-inflammatory, methotrexate, sulfosalazina, other) can be singled out.
Curious Image from bone marrow
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The CBC test results for some patients show a mild to a moderate decline in MCV, in spite of the normal levels of Hb, HCT, and RBC count, indicating there is microcytosis of erythrocytes without anemia. We see this pattern of CBC results more frequently for young males, some of them showing fairly high Hb concentration. What is your practical approach to deal with these results? 
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In the absence of iron deficiency and anaemia of chronic disease, thalassaemia trait (heterozygous beta thalasaemia or alpha thalassaemia due to single / 2 gene deletion) is most likely. A simple formula known as the Mentzer index will be able to distinguish iron deficiency from thalassaemia trait. Divide the MCV by the red cell count: an MCV/RCC ratio <13 is suggestive of thalassaemia trait. To distinguish between alpha and bata thal trait, do haemoglobin sub-fractionation studies (HPLC or Hb electrophoresis). An elevated HbA2 level would be in keeping with beta thalassaemia trait. In Alpha thalassaemia trait the HbA2 level is either normal or decreased. 
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Why not the other size?
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Biconcave shape mostly help to squeeze RBC through the narrow capillaries.
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I request you to please share your research, views, and suggestions. 
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Sure
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I want to count leukocytes in the blood circulation.
My problem is that the cell number varies a lot. Which method is the most reliable method to count blood cells from mice?
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What about counting rbc in mice using haemocytometer?
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Can someone please provide the reference sequence for the coagulation factor 8 gene? It is located on the long arm of the X chromosome (X-q28). I am trying to allign some primers to detect inversions in intron 22 and intron 1 which are responsible for the development of severe haemophilia A.
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Hi Nicolas
you can also get all what you want on the UCSC website. take care when designing primers since this gene has a pseudogene with high homologies. therefore test your primers by an in silico PCR, or choose them in the non homologue regions (blast the F8 sequence in UCSC, you'll see what differs between both genes).
fred
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I would like someone to help me in order to analyze length telomere. I followed the lkit instructions but I do not get a good separation between cell populations by flow cytometry.
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We have used this kit years ago when it first came out, have spent quite a time to standardise it, you need the to read the instructions carefully and do the pipetting correctly. when you do washes, to get rid of all of the supernatant is important.Our tech used to pipet out the remnants of the supernatant after centrifugation which made the difference for clear cut populations..
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We are working with C57BL/6J Apc min mice but even a reference just for C57BL/6J mice would be great!
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Hi Janay, since the C57Bl6 strain is so heavily interbred, I would expect that there would be a reference for their blood components.
But, I cant find one.
I would suggest that you contact the mouse suppliers and ask them if they did hematology measurements of their populations.
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When I incubated opsonized erythrocytes and monocytes (monocytes and erythrocytes from the same human donor), the phagocytosis level was lower (11.8% of ingesting erythrocytes), than when I incubated monocytes and opsonized erythrocytes from different human donors (26.64% ingesting erythrocytes). When I incubated nonopsonized erythrocytes (uKKcz) with monocytes (used 24h after isolation) the phagocytosis level was low (0.2% intgesting erythrocytes). When I incubated nonopsonized eryhrocytes (uKKcz) with monocytes (used immediately after isolation) the phagocytosis level was higher (7.46% ingesting erythrocytes). Does anyone know how to interpret my research results of erythrophagocytosis? Any suggestions?
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1. Opsonized erythrocytes and monocytes  - 11.8% of ingesting erythrocytes
2. Monocytes and opsonized erythrocytes - 26.64% ingesting erythrocytes
3. Nonopsonized erythrocytes (uKKcz) with monocytes - 0.2% intgesting erythrocytes
4. Nonopsonized eryhrocytes (uKKcz) with monocytes -  7.46% ingesting erythrocytes
Interpretations : Opsonized erythrocytes are more prone to get phagocytosed by monocytes.
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Can anyone suggest why CD47 expression was higher in old RBCs concentrates with leukocytes than in RBCs concentrates without leukocytes?
(leukodepleted)?
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First , recheck the calibration of your machines. Then , try to optimize the CD47 expression with different concentration  of EBC : WBC ratio and then get it confirmed. 
As someone told before, macrophage tend to phagocytose the RBC , so that RBC with Macrophage need to express more " don't eat signal " cd47 to prevent pahocytosis by macrophage. 
Though , nowadays, a new complex of CD47 - SIRPa are seen to initiate phagocytosis.
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When I analyze CSF by DxH Beckmann Coulter, regularly get higher RBC count in comparison with manually counted RBC. TNC values are comparable in both way of counting.
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