204 Blood Analysis of Cas9-Expressing Cattle

2018 ◽  
Vol 30 (1) ◽  
pp. 242
Author(s):  
S.-Y. Yum ◽  
S.-J. Lee ◽  
S.-E. Hahn ◽  
C.-I. Lee ◽  
H.-S. Kim ◽  
...  
Keyword(s):  
Blood Cells ◽  
Gene Editing ◽  
White Blood Cells ◽  
Blood Analysis ◽  
Health Issues ◽  
Blood Samples ◽  
Vitro Fertilization ◽  
Transgenic Cattle ◽  
Near Future

The CRISPR/Cas9 system has proved to be a powerful tool for knockout and knock-in in various species. When 2 components—Cas9 and single guide (sg)RNA—are delivered into cells or embryos, the events of gene editing occur. Because Cas9 is essential for gene editing in the CRISPR/Cas9 system, some studies have reported the production of Cas9-expressing animals, such as mice, which could be used to increase gene editing efficiency in subsequent experiments. In previous reports, we successfully produced 4 Cas9-expressing cattle via microinjection (Hahn et al. 2016 Reprod. Fertil. Dev. 29, 211). Primary cells from these calves had Cas9 activity because transfection of only sgRNA resulted in gene deletion. The aim of this study was to analyse the blood of the transgenic cattle to investigate the effect of Cas9 expression on health. Two of 4 transgenic calves died; one had severe ruminant tympany, failed to respond to treatment, and died at 4 months of age, and the other died at 5 months of age due to accidental ingestion of a needle from a feed bunk. Blood samples were obtained from the surviving 2 transgenic cattle (1 male and 1 female) at 7 and 12 months for blood analysis. Five milliliters of whole blood samples was collected from the jugular vein. Portions were used for CBC (Hemavet 950, Drew Scientific, Miami Lakes, FL, USA) and for serum chemistry analysis (BS-400, Mindray, Shenzhen, China). Average values for white blood cells (9600 and 1057/mm3), neutrophils (4590 and 3870/mm3), lymphocytes (4020 and 5910/mm3), red blood cells (732,000 and 798,000/mm3), hemoglobin (9.5 and 10.2 g dL−1), packed cell volume (24.3 and 25.3%), platelet (439,000 and 327,500/mm3), AST (76 and 104 IU), ALP (140 and 133 IU), BUN (7.5 and 10.5 mg dL−1), and creatinine (1.3 and 1.0 mg dL−1) of male and female transgenic calves were within the reference range. Additionally, there was no difference in general health information, including body temperature and feeding. In conclusion, we demonstrated that continuous Cas9 expression in transgenic cattle did not affect health status of the surviving calves in terms of blood analysis. They have grown up without any health issues and are currently 14 (female) and 15 (male) months old. In the near future, we will evaluate their germline transmission by natural breeding or in vitro fertilization. This work was supported by BK21 PLUS Program for Creative Veterinary Science, NRF (NRF-2017R1A2B3004972), and Seoul Milk Coop (SNU 550–20160004).

scholarly journals Moderate Range Static Magnetic Field Promoted Variation of Blood Parameters: An In vitro Study

2020 ◽  
Vol 8 (1) ◽  
pp. 55-64
Author(s):  
Bestoon T. Mustafa ◽  
Sardar P. Yaba ◽  
Asaad H. Ismail
Keyword(s):  
Magnetic Field ◽  
Blood Cell ◽  
Static Magnetic Field ◽  
Blood Cells ◽  
White Blood Cells ◽  
Albino Rats ◽  
Field Exposure ◽  
Blood Samples ◽  
Cell Counts

This study was undertaken to investigate the influence of a homogenous and uniform static magnetic field (SMF) on the main blood cell counts in vitro experiment. Fresh blood samples were collected from albino rats and exposed to SMF (2.4, 6, 25, 50, 75, and 100 mT) versus 15–60 min. Results showed a significant change of blood counts under the low field effects. A 2.4 mT was a trend of white blood cells (WBCs) count increase non-linearly. However, a 6 mT exposure reduced WBCs with about 39%. Other variations fluctuated within 30%. The 25 mT decreased red blood cells (RBCs), hemoglobin, and hematocrit levels with 13% similarly. The lower exposure field, (2.4 and 6) mT, and effects on RBCs were 6% fluctuation. The 6 mT reduced platelet counts with half in comparison to control blood samples. About 20% increase obtained due to 50 mT exposure during all period. None of 75 and 100 mT exposures dominated blood counts alterations. The quiet magnetic field exposure for a certain time can be interesting to control blood cell count-related diseases.

scholarly journals Shorter telomere length of white blood cells is associated with higher rates of aneuploidy in women undergoing in vitro fertilization

2019 ◽  
Vol 112 (3) ◽  
pp. e76
Author(s):  
Brent M. Hanson ◽  
Xin Tao ◽  
Yiping Zhan ◽  
Julia G. Kim ◽  
Emily K. Osman ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Telomere Length ◽  
Blood Cells ◽  
White Blood Cells ◽  
Vitro Fertilization

NORMAL GRANULOCYTE INFUSION THERAPY FOR ASPERGILLOSIS IN CHRONIC GRANULOMATOUS DISEASE

PEDIATRICS ◽  
1973 ◽  
Vol 51 (2) ◽  
pp. 230-233
Author(s):  
Andrew A. Raubitschek ◽  
Alan S. Levin ◽  
Daniel P. Stites ◽  
Edward B. Shaw ◽  
H. Hugh Fudenberg
Keyword(s):  
Adverse Effects ◽  
Chronic Granulomatous Disease ◽  
Blood Cells ◽  
White Blood Cells ◽  
Infusion Therapy ◽  
Granulomatous Disease ◽  
Transient Improvement ◽  
Life Threatening ◽  
Granulocyte Function

An 8-year-old boy with chronic granulomatous disease (CGD) was admitted in moribund condition with aspergillus pneumonia. Because of the gravity of the situation, normal granulocyte infusions were used as adjuncts to the more conventional antimicrobial therapy. White blood cells, derived from a total of 58 units of whole blood obtained by leukophoresis of the father, were given in two separate doses. The first dose, totaling 2.8 x 1010 granulocytes, was coincident with significant improvement, and the second, totaling 3.0 x 1010 granulocytes, was coincident with the onset of clinical improvement and interim recovery. Transient improvement in in vitro granulocyte function was noted in cells taken from the patient's blood immediately after infusion. No adverse effects of the infusions were noted in either the patient or the donor. Although it is impossible to divorce the therapeutic effect of the granulocyte infusions from the more conventional therapy, we conclude that normal granulocyte infusions can be considered a valid adjunct in children with CGD who are suffering from a life-threatening infection.

Utility of 8 h and time decay-corrected acquisition scintigraphy with in-vitro labeled white blood cells for the diagnosis of osteoarticular infection

2017 ◽  
Vol 38 (6) ◽  
pp. 500-508 ◽  
Author(s):  
Edel Noriega-Álvarez ◽  
Guillermo A. Martínez Pimienta ◽  
Ana M. Benítez Segura ◽  
María T. Bajén Lázaro ◽  
Alba Rodríguez-Gasén ◽  
...  
Keyword(s):  
Blood Cells ◽  
White Blood Cells ◽  
Time Decay ◽  
Osteoarticular Infection

EFFECTS OF BLOOD CELLS ON PLATELET AGGREGATION BY IMPEDANCE METHOD

1987 ◽  
Author(s):  
L Mannucci ◽  
R Redaelli ◽  
E Tremoll
Keyword(s):  
Platelet Aggregation ◽  
Whole Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Normal Subjects ◽  
Impedance Method ◽  
Induced Aggregation ◽  
Vitro Data ◽  
In Vitro Data

To evaluate the effects of blood cells on the response of platelets to aggregating agents using whole blood impedance aggregometer, studies were carried out on whole blood (WB) of normal subjects and of patients with: polycythemia vera (PV), iatrogenic anemia (IA), primary thrombocytosis (PT), idiopathic thrombotic purpura (ITP), myeloid chronic leukemia (MCL), iatrogenic leukopenia (IL). The in vitro effects of red blood cells (RBC) and of white blood cells (WBC) on platelet rich plasma (PRP) aggregation were also evaluated. WB, PRP, WBC and RBC were prepared by conventional methods. Aggregation was performed using the impedance aggregometer (mod. 540, Chrono Log Corp). In normal subjects the concentration of collagen giving 50 % aggregation (AC50 ) found in PRP did not differ from that of WB, indicating that hematocrit values within the normal range did not appreciably affect platelet aggregation. The results obtained in WB of patients are summarized in the table: In vitro data showed that aggregation in prp in wb of normal subjects was related to the number of platelets present in the sample. RBC added to PRP significant reduced aggregation only when the RBC number was greater than 4.101 cells. No effect of WBC on collagen induced aggregation of PRP was observed, whereas significant inhibition was detected after ADP. It is concluded that the aggregation evaluated in WB with impedance method is dependent on the platelet number. Also, in vitro data and studies in WB of patients indicate that aggregation is significantly affected by the presence of cells other than platelets only in conditions of changes of the ratio between platelets and leukocytes and/or red cells.

scholarly journals Acetylsalicylic acid and morphology of red blood cells

2010 ◽  
Vol 53 (3) ◽  
pp. 575-582 ◽  
Author(s):  
Jacques Natan Grinapel Frydman ◽  
Adenilson de Souza da Fonseca ◽  
Vanessa Câmara da Rocha ◽  
Monica Oliveira Benarroz ◽  
Gabrielle de Souza Rocha ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Control Group ◽  
Blood Samples ◽  
Morphological Alterations ◽  
Blood Smears ◽  
In Vivo Treatment ◽  
Microscopy Data

This work evaluated the effect of in vitro and in vivo treatment with ASA on the morphology of the red blood cells. Blood samples or Wistar rats were treated with ASA for one hour. Blood samples or animals treated with saline were used as control group. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphology of red blood cells were evaluated under optical microscopy. Data showed that the in vitro treatment for one hour with ASA at higher dose used significantly (p<0.05) modified the perimeter/area ratio of the red blood cells. No morphological alterations were obtained with the in vivo treatment. ASA use at highest doses could interfere on shape of red blood cells.

scholarly journals Comparison of the activation capabilities of hemosorbents by adhesion rate of blood cells in vitro

2021 ◽  
Vol 8 (5) ◽  
pp. 57-66
Author(s):  
S. I. Kuznetsov ◽  
O. P. Kirichuk ◽  
N. V. Burkova ◽  
G. O. Yuriev ◽  
V. A. Davankov ◽  
...  
Keyword(s):  
Clinical Practice ◽  
Blood Cells ◽  
Solid Phase ◽  
Activation Functions ◽  
Activation Effect ◽  
Blood Samples ◽  
Adhesion Rate ◽  
Low Volume ◽  
Donated Blood

Background: The relevance of the work lies in the search for new hemocontact drugs with hemocompatibility and a pronounced activation effect on the cellular and humoral blood systems for their possible use in clinical practice during low-volume hemoperfusion.The aim of this work was to assess the activation capabilities of three granular hemosorbents by the rate of adhesion of blood cellular elements to the surface of granules in vitro.Materials and methods. When using the method of low-volume hemoperfusion (LVH) in the clinic it is important to take into account the activation properties of solid-phase granular drugs. Blood-contact interaction was carried out in bench conditions with the use of donated blood in rotary mode. Blood samples were taken before the experiment and after 5, 20, 40 and 60 minutes. Changes in blood cell and subcellular populations were evaluated using the Sysmex XT 1800i hematological analyzer (26 parameters), which made it possible to indirectly judge the activation of blood cells. 30 experiments were conducted. To analyze the activation functions of the hemocontact preparations the speed-time adhesive profile of blood cells on the sorbent was used.Results. The effect of using the preparations Silochrome S-120 and SPS in comparison with SСT-6A HP as contact hemoactivators can be more pronounced, since the activation potential of these sorbents for blood cells is much higher. Silochrome S-120 has the highest activation capabilities compared to SPS and SKT-6A HP.Conclusion. Adhesion rate indicators can be indicators of the activation of blood cells upon contact with foreign surfaces and serve as a criterion for assessing the activation capabilities of these surfaces when using the LVH method in the clinic.

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