Serum variants causing the formation of double hearts and other abnormalities in explanted rat embryos

Development ◽  
1974 ◽  
Vol 31 (3) ◽  
pp. 707-719
Author(s):  
C. E. Steele ◽  
D. A. T. New
Keyword(s):  
Embryonic Development ◽  
Blood Cells ◽  
Blood Clot ◽  
Fibrin Clot ◽  
Normal Blood ◽  
Rat Embryos ◽  
Homologous Serum

Rat embryos explanted before organogenesis (8½ days gestation) were grown in culture in homologous serum. When the serum was prepared from blood centrifuged after clotting, the embryos developed double hearts. In serum prepared from blood centrifuged before clotting had occurred, and in plasma, the embryos developed normal single hearts. The delayed-centrifuged (D.C.) serum also supported less growth of older embryos than the immediately-centrifuged (I.C.) serum. The harmful properties of D.C. serum appeared rapidly in contact with a normal blood clot but did not develop in contact with separated blood cells and fibrin clot. Mixtures of D.C. and I.C. sera gave results intermediate between those from the two sera alone. No significant differences were found between D.C. and I.C. serum in calcium or complement content but both supported better embryonic development after pre-heating.

scholarly journals Contribution of Red Blood Cells and Clot Structure to Thrombosis

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. SCI-15-SCI-15
Author(s):  
Robert A S Ariens
Keyword(s):  
Atomic Force Microscopy ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Splice Variant ◽  
Blood Clot ◽  
Fibrin Clot ◽  
Force Microscopy ◽  
Atomic Force ◽  
Fibrin Network ◽  
Clot Structure

Abstract The blood clot or thrombus is composed of red blood cells, platelets and white cells that interact with the fibrin meshwork produced by the coagulation system. The red blood cells form polyhedrocytes that seal the clot, platelets provide forces for clot contraction, and white cells contribute neutrophil extracellular traps, cytokines and complement activation. The fibrin network contributes clot elasticity and provides the proteinaceous backbone structure that stabilises the clot. Previous studies from our laboratory and others have shown that dense fibrin networks demonstrating small pores and increased resistance to fibrinolysis associate with thrombosis. However, the mechanisms underpinning this have not been fully understood. Recent studies using atomic force microscopy from our laboratory have shown that fibrinogen interacts with red cells with comparable affinity as that with platelets. A patient with mutations in the β3 integrin subunit showed no binding between red cells and fibrinogen, demonstrating that a β3-related integrin receptor is involved in the interaction. Mutations in the fibrinogen α-chain integrin binding sites (D97E and D574E) reduced frequency of red cell interactions with fibrinogen. Interestingly, a naturally occuring splice variant of the fibrinogen γ-chain that reduces binding to the platelets, fibrinogen γ', increased binding interactions between fibrinogen and red blood cells. Fibrinogen γ' is a naturally occurring splice variant of fibrinogen, in which the C-terminal AGDV residues of the more common γA-chain (85%) are replaced with a negatively charched VRPEHPAETEYDSLYPEDDL sequence of the γ' chain (15%). Fibrinogen γ' induced clustering of fibrin fibres into tightly interknit nuclei of fibrin fibres, interspersed by large pores that extend over more than 50 μm within the fibrin network structure. The effects of fibrinogen γ' on fibrin clot structure was independent of thrombin and FXIII as demonstrated using snake venom enzyme. Previously we showed impaired fibrin protofibril formation with fibrinogen γ' using atomic force microscopy. Using turbidimetric analysis of fibrin intrafibrillar structure, we show that fibrinogen γ' reduces protofibil packing per fibrin fiber. Furthermore, we find that reduced protofibril packing diminishes fibrin stiffness as analysed with magnetic tweezers both in purified systems as well as in plasma at (patho)physiological fibrinogen γ' levels that range from 3-40%, and in whole blood as analysed with thromboelastography. In conclusion, our data show that red blood cells and fibrinogen γ' play major roles in the regulation of clot structure and stability, and that these effects on clot structure are major determinants of the functional properties of the blood clot. Modulating fibrin clot structure and its interactions with blood cells may represent major new targets for the treatment of thrombosis. Disclosures No relevant conflicts of interest to declare.

Experimental studies in insect parasitism XIV. The haemocytic reaction of a caterpillar to larvae of its habitual parasite

1966 ◽  
Vol 165 (999) ◽  
pp. 155-178 ◽  
Keyword(s):  
Foreign Body ◽  
Embryonic Development ◽  
Chemical Treatment ◽  
Blood Cells ◽  
Protective Property ◽  
Foreign Bodies ◽  
Experimental Studies ◽  
Ephestia Kuehniella ◽  
Main Theme ◽  
Ionic Exchange

Although caterpillars of Ephestia kuehniella promptly encapsulate alien parasites and other foreign bodies in their haemocoele, they do not normally encapsulate larvae of their habitual parasite Nemeritis canescens , which develop unhindered and eventually destroy their host. The larva of Nemeritis does not achieve this immunity by repelling the blood cells, or by physically dislodging them. It is immune because it is able to live in the haemocoele of Ephestia without evoking a haemocytic reaction; presumably, that is, because it is not recognized as a foreign body. That ability is due to a property of its surface. So long as its surface remains unaltered, the larva, alive or dead, evokes no haemocytic reaction. When its surface is altered whether by perforation, abrasion, or chemical treatment, the living larva evokes a haemocytic reaction in Ephestia and becomes encapsulated. The protective property of its surface is acquired by the larva very late in its embryonic development, between 62 and 66 hours of age at 25 °C. This is about the same time as, or a little later than, the cuticle of the embryonic larva becomes impermeable to water. Four fat solvents were found to deprive the living larva of its immunity, but they may have affected the protective surface by disrupting the underlying wax layer of the epicuticle. Treatments and substances that did not affect the protective surface give some crude indications of its properties, but its ultimate characterization must be in terms of insect immunology. Observations incidental to the main theme of the paper show that the cuticle of the larva is impermeable to water; that ionic exchange takes place through the anus and wall of the rectum, where some food substances may also be absorbed from the blood of the host; and that the order of formation of the cuticulin and wax layers of the embryonic larva is the same as that in ecdysis from instar to instar in other insects. They also provide information on the longevity of bitten supernumerary larvae.

Abstract TP35: Estimation of Red Blood Cells in the Thrombus Using MRI. A Phantom Study with Predetermined Thrombus Components

Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Ana Paula Narata ◽  
Isabelle Filipiak ◽  
Richard Bibi ◽  
Jean Philippe Cottier ◽  
Kevin Janot
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Spin Echo ◽  
In Vitro Study ◽  
Fibrin Clot ◽  
Fluid Attenuated Inversion Recovery ◽  
Magnetic Resonance Imaging Mri ◽  
C 50 ◽  
D 20

Background and Purpose: Better understanding about thrombus composition seems necessary, as treatment of acute ischemic stroke (AIS) is focus on clot chemical dissolution and mechanical extraction. We propose to evaluate whether magnetic resonance imaging (MRI) can differentiate white from red clots and estimate red blood cells percentage (RBC%) using clots with predetermined components and an index based on MRI signal intensity (SI). Material and Methods: 5 clots (A=100% fibrin, B=80% RBC, C=50% RBC, D=20% RBC, E=unknown) were fixed in gelatin-manganese solution and studied by: high-resolution 3D T1-weighted (T1MPR), T2-weighted turbo spin echo (T2TSE), T2-weighted gradient echo (T2GE), susceptibility weighted (SWI), fluid-attenuated inversion recovery (FLAIR) and diffusion weighted imaging (DWI) with apparent diffusion coefficient (ADC). SI index was calculated with clot SI and gelatin SI. Statistical analysis compared RBC-clots to fibrin-clot SI index and the correlation of RBC% and SI index in each MRI sequence. Results: Each red clot was different from clot A except clot D in FLAIR. Correlation between clots SI index and RBC concentration were found in T1MPR (r=-0.84), SWI (r=-0.79), T2GE (r=-0.72) and FLAIR (r=0.80). Linear regression resolution provided an indirect RBC estimation for clot E: 47.3 % in T1MPR, SWI 41.5%, T2GE 45.1% and FLAIR 50.9%. Histological analysis confirmed clot E composition. Conclusion: This in vitro study suggests that MRI can differentiate white from red clots except clots with low RBC% in FLAIR and also provide approximate RBC%.

In vitro analysis of glucose metabolism and embryonic growth in postimplantation rat embryos

Development ◽  
1987 ◽  
Vol 100 (3) ◽  
pp. 431-439 ◽  
Author(s):  
S.K. Ellington
Keyword(s):  
Glucose Metabolism ◽  
Embryonic Development ◽  
Glucose Uptake ◽  
Glucose Concentration ◽  
Culture Media ◽  
Rat Embryos ◽  
Embryonic Growth ◽  
Stage Of Development

The glucose metabolism and embryonic development of rat embryos during organogenesis was studied using embryo culture. Glucose uptake and embryonic growth and differentiation of 10.5-day explants (embryos + membranes) were limited by the decreasing glucose concentration, but not the increasing concentration of metabolites, in the culture media during the second 24 h of a 48 h culture. No such limitations were found on the embryonic development of 9.5-day explants during a 48 h culture although glucose uptake was slightly reduced at very low concentrations of glucose. From the head-fold stage to the 25-somite stage of development, glucose uptake was characteristic of the stage of development of the embryo and not the time it had been in culture. Embryonic growth of 9.5-day explants was similar to that previously observed in vivo. Glucose uptake by 9.5-day explants was dependent on the surface area of the yolk sac and was independent of the glucose concentration in the culture media (within the range of 9.4 to 2.5 mM). The proportion of glucose converted to lactate was 100% during the first 42h of culture then fell to about 50% during the final 6h. The protein contents of both the extraembryonic membranes and the embryo were dependent on the glucose uptake.

scholarly journals The Chile Biliary Longitudinal Study: A Gallstone Cohort

2020 ◽  
Author(s):  
Jill Koshiol ◽  
Vanessa Van De Wyngard ◽  
Emma E McGee ◽  
Paz Cook ◽  
Ruth M Pfeiffer ◽  
...  
Keyword(s):  
Longitudinal Study ◽  
Blood Cells ◽  
Blood Clot ◽  
Buffy Coat ◽  
Central Chile ◽  
Southern Central ◽  
Risk Areas ◽  
Detection Strategies ◽  
Primary Risk Factor

Abstract Gallbladder cancer (GBC) is a highly fatal cancer that can be cured through cholecystectomy if identified early. The presence of gallstones is the primary risk factor for GBC, but few people with gallstones develop GBC. A key question is what drives the development of GBC among persons with gallstones. We initiated the Chile Biliary Longitudinal Study (Chile BiLS) to address this question. From 2016 to 2019, Chile BiLS enrolled 4,726 women aged 50–74 years with ultrasound-detected gallstones from southern-central Chile, accounting for an estimated 36% of eligible women with gallstones in the study area. The median age was 59 years; 25% of the women were Amerindian (Mapuche), 60% were obese, 25% had diabetes, and 6% had cardiovascular disease. Participants will be followed for gallbladder dysplasia or cancer for 6 years. As of April 30, 2020, over 91% of those eligible completed the year 2 follow-up visit. Data being collected include epidemiologic and sociodemographic information, anthropometric measurements, blood pressure, and tooth counts. Biosamples being taken include baseline plasma, buffy coat, red blood cells, serum, blood clot from serum, and PAXgene whole blood (PreAnalytiX GmbH, Hombrechtikon, Switzerland). Complete gallbladder sampling is conducted for most participants undergoing cholecystectomy. The Chile BiLS cohort study will increase our understanding of GBC etiology and could identify potential risk stratification and early detection strategies in high-risk areas.

scholarly journals Flow cytometric analysis of Notch1 and Jagged1 expression in normal blood cells and leukemia cells

2012 ◽  
Vol 4 (3) ◽  
pp. 397-400 ◽  
Author(s):  
ERIKO KANAMORI ◽  
MAI ITOH ◽  
NAOKO TOJO ◽  
TAKATOSHI KOYAMA ◽  
NOBUO NARA ◽  
...  
Keyword(s):  
Blood Cells ◽  
Flow Cytometric Analysis ◽  
Leukemia Cells ◽  
Normal Blood ◽  
Flow Cytometric

The Y-Specific Gene PRY Is Expressed in Normal Blood Cells as Well as Leukemia Cells and Can Elicit a Specific Antibody Response in Male Recipients of Hematopoietic Stem Cells from Female Donors.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4976-4976
Author(s):  
Emmanuel Zorn ◽  
Blair Floyd ◽  
Andrew Tweel ◽  
David B. Miklos ◽  
Roberto Bellucci ◽  
...  
Keyword(s):  
B Cell ◽  
Cell Lines ◽  
Blood Cells ◽  
Leukemia Cell ◽  
Normal Blood ◽  
Specific Gene ◽  
Hematopoietic Stem ◽  
Allogeneic Hsct ◽  
Leukemia Cell Lines ◽  
Male Patients

Abstract H-Y minor histocompatibility antigens (mHA) are common targets of immune responses following allogeneic hematopoietic stem cell transplantation (HSCT) in male patients who receive stem cells from female donors. These H-Y antigens are encoded by a group of genes located on the non-recombining portion of the Y chromosome. H-Y genes are ubiquitously expressed with 1 to 13% disparity at the protein level with their X homologues. The same portion of the Y chromosome also contains a distinct group of 11 Y-specific genes, for which there are no X homologues. Expression of Y-specific genes is reportedly restricted to the testis thereby limiting the potential relevance of these proteins as immune targets following allogeneic HSCT. However, atypical expression of Y-specific genes has recently been reported in prostate cancer, suggesting that these genes might follow a pattern of expression characteristic of cancer-testis antigens. In this study, we investigated the expression of a representative Y-specific gene, PRY, in male leukemia cell lines, and examined the immunogenicity of this protein in male patients who received allogeneic HSCT from female donors. Using DNAse treated RNA in RT-PCR experiments we showed that PRY gene is expressed in 3 of 6 male leukemia cell lines tested but not in 4 female cell lines. Results were confirmed by southern blotting of PCR products using an internal specific probe. We next assessed PRY expression in normal blood cells collected from 3 male and 3 female donors. In contrast to what has previously been reported, PRY gene was found expressed at low levels in blood cells from all male donors but not from female donors. Although the precise phenotype of cells expressing PRY remains unknown, expression in normal hematopoietic as well as tumor cells suggested that Y-specific gene products could also elicit immune responses after sex mismatched allogeneic HSCT. We used a series of overlapping peptides encompassing the entire sequence of PRY in ELISA assays to examine the antibody response to PRY antigen in male recipients of female transplant. Thus far, 1 of 13 serum samples has been positive for antibody to PRY. This sample, collected approximately one year post-transplant, strongly reacted with a single peptide, indicating that the patient had developed a B cell response to the PRY antigen. This patient with AML had received a HSCT from his HLA identical female sibling. After transplant, he developed acute and chronic GVHD. Remarkably, this patient also developed a strong CD4+ T cell and B cell response to another H-Y antigen, DBY, following HSCT. Studies are ongoing to further characterize the immune response elicited towards this newly defined antigen. Overall, our data indicate that the Y-specific gene PRY is expressed in normal blood cells as well as leukemia cells. Following allogeneic HSCT, PRY can also trigger B cell immunity in male patients with female donors. Further studies are required to determine whether other Y-specific gene products are also immunogenic and constitute a new category of mHA with significant implications in the development of GVL and GVH reactions.

af9 Regulates gata2 Expression During Early Hemangioblast Specification and Vascular Pattern Formation In Zebrafish.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2600-2600
Author(s):  
Giuseppe Germano ◽  
Ilaria Guariento ◽  
Natascia Tiso ◽  
Blaine W. Robinson ◽  
Enrico Moro ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Pattern Formation ◽  
Embryonic Development ◽  
Blood Cells ◽  
Erythroid Cell ◽  
Fusion Partner ◽  
Vascular Pattern ◽  
Cell Stage ◽  
Whole Mount

Abstract Abstract 2600 Background AF9 is a transcription factor that plays an essential role in hematopoiesis and embryonic development. The alteration of AF9 is principally associated in acute myeloid leukemia as fusion partner of human MLL (mixed-lineage leukemia) gene rearrangements. Zebrafish is an excellent model organism to study embryonic development and hematopoiesis. We have previously shown that zebrafish af9 is expressed within the intermediate cell mass (ICM), a site of primitive hematopoiesis in zebrafish. Here we study the loss of af9 in zebrafish development to further understand how af9 modulates early hematopoietic and embryonic development. Methods and results Two morpholino antisense oligos (MOs), designed to block translation and inhibit pre-mRNA splicing of af9, were co-injected in embryos at 1–2 cell stage. To control for off-target effects, two morpholino mismatch oligos were designed and co-injected. Efficacy of MOs was demonstrated by Western blot analysis and RT-PCR in controls and MO-injected embryos (morphants). In vivo monitoring of both morphants and control embryos was carried out by microscopy. Effects of af9 depletion on vasculature and erythropoiesis were evaluated in Tg(fli1:eGFP) and Tg(gata1:DsRed) transgenic lines, respectively. Whole-mount in situ hybridization of known hematopoietic markers was used to decipher the developmental time-points in which af9 regulates blood development. Following injection of two MOs at 1–2 cell stage, we compared the morphological features of the morphants with control embryos at about 24 hours post-fertilization (hpf). The af9 morphants showed small head and eyes, disruption of tail development and pronounced swelling in the posterior ICM. Circulating blood cells were reduced from 26 hpf to later stages of development. At 48 hpf the heart was enlarged, showed a paucity of blood-cells and pericardial edema. Decreased number of blood cells in morphant embryos was further confirmed by o-dianisidine staining at 48 hpf and 72 hpf and in living af9-knockdown gata1:DsRed transgenic animals, suggesting that the differentiation of erythroblasts remains insufficient or impaired. Concordant with this observation, we examined the expression of specific markers for early hematopoiesis (scl, lmo2 and gata2) and primitive erythropoiesis (gata1, hbbe, and band3) using whole-mount in situ hybridization (WISH). At the 5-somite stage, the early hematopoietic precursor marker gata2 was markedly increased while scl and lmo2 remained unaffected in af9 morphants. Interestingly, by 24 hpf gata2 was found to be specifically over-expressed in ICM while no change was observed for scl and lmo2 markers. Besides, the erythroid progenitors and mature erythrocyte markers gata1, band3 and hbbe displayed nearly normal expression. To further confirm the role af9 in early hematopoiesis, we examined its expression in moonshine, a mutant zebrafish with defects in erythroid maturation due to deficiency of tif1γ, a key regulator of hematopoietic gene expression. WISH analysis in moonshine showed loss of af9 expression in the ICM at 24 hpf, suggesting that af9 functions genetically downstream of tif1γ in normal erythroid cell development. To determine the effect of af9 on endothelial and vascular development, we performed knockdown of af9 in fli1:eGFP transgenic line. By 24 hpf, these morphants showed significant increase of fluorescence intensity in the posterior ICM and a clear perturbation in the inter-segmental vessels (ISV) of the trunk at 30 hpf, indicating that af9 is required for early steps in hemangioblast specification and vascular pattern formation in zebrafish. Conclusion af9 regulates gata2 expression during early hemangioblast specification and vascular pattern formation in zebrafish. af9 may also be involved in caudal segment morphogenesis. Taken together, these data provide the initial framework of a pathway that can be used to further integrate the molecular events regulating hemangioblast differentiation. Disclosures: No relevant conflicts of interest to declare.

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