X‐ and gamma‐irradiation have similar effects on the in vitro quality of stored red cell components

Transfusion ◽  
2021 ◽  
Author(s):  
Denese C. Marks ◽  
Rachel G. Webb ◽  
Claire Linnane ◽  
Htet Htet Aung ◽  
Peta M. Dennington ◽  
...  
2020 ◽  
Vol 47 (5) ◽  
pp. 409-419
Author(s):  
Nitesh Gupta ◽  
Dheeraj Khetan ◽  
Rajendra Chaudhary ◽  
Jai Shankar Shukla

Introduction: Damage to a cell and the loss of integrity of its cell membrane leads to the release of endogenous immunogenic molecules, which together are classified as “damage-associated molecular patterns” (DAMPs). Cell-free DNA (cf-DNA) released from nucleosomes may serve as a proco­agulant cofactor and may be an important mediator of immunomodulatory and proinflammatory effects associated with blood transfusion. Objectives: To assess the levels of cf-DNA in supernatants of stored red cell components and the effect of leukoreduction and gamma irradiation on the release of cf-DNA during storage. Methods: This is a prospective cohort study on 99 stored red cell components, randomly divided into three groups – buffy coat (BC)-depleted, leuko-filtered (LP), and irradiated (IR) packed red blood cells. Red cell supernatants were drawn over a period of 21 days at three different time points (day 0, 7, and 21) from the study units. cf-DNA extraction was done and quantified by a bench top fluorometer. Change in cf-DNA content, rate of change (μg/day), and percent change were estimated and compared across different groups. Results: cf-DNA content increased (p = 0.000) with storage duration in the BC (median = 238.66 μg, interquartile range [IQR] = 168.42 on day 21 vs. median = 9.44 μg, IQR = 5.23 on day 0) and IR groups (p = 0.000) (median = 245.55 μg, IQR = 253.88 on day 21 vs. median = 7.07 μg, IQR = 13.58 on day 0), while there was a decreasing trend (p = 0.032) in the LP group (median = 4.55 μg, IQR = 10.73 on day 21 vs. median = 8.66 μg, IQR = 6.56 on day 0). The median rate of change in cf-DNA content (11.13 μg/day) and percent change in cf-DNA content (median = 4,106.16%) was highest in the IR group. Conclusions: Stored red cell components contain significant amount of cf-DNA. Release of cf-DNA is further aggravated by irradiation while leukoreduction leads to a decrease in cf-DNA content.


2016 ◽  
Vol 10 (3) ◽  
pp. 355
Author(s):  
Yuyu S Poerba ◽  
MARIA IMELDA ◽  
AIDA WULANSARI ◽  
DIYAH MARTANTI

Amorphophallus muelleri Blume (Araceae) is valued for its glucoman content for use infood industry (heathy diet food), paper industry, pharmacy and cosmetics. The cultivationof A. muelleri is hampered by limited genetic quality of seed. The species is triploid(2n=3x=39), the seed is developed apomictically, and pollen production is low. Thespecies is only propagated vegetatively. This may explain that the species is difficultto breed conventionally and genetic variability in the existing landraces cultivars israther limited. Conservation of this species, therefore, is important for availability of thespecies in the future use of this plant. The objective of present research is to increasegenetic variation by induce mutation using gamma-rays irradiation of shoot culturesof A. muelleri and to identify DNA polymorphism induced by gamma irradiation usingrandom amplified polymorphic DNA (RAPD), so the mutants produced can be used forbreeding purposes and for conservation program. Results of the experiment showedthat gamma irradiation less than 5 gray was effective to induce mutation of A. muelleri.Four RAPD primers generated 35 scorable bands with 100% polymorphic bands. Sizeof the bands varied from 350bp to 2.0kbp. Clustering analysis was performed based onRAPD profiles using the UPGMA method. The range of genetic distance among individualgenotypes was from from 0.00 to 0.72, while genetic variance of the population was0.21 + 0.13. The eighteen genotypes were proof to be mutants. The mutants producedin this experiment could be used as new germplasms for breeding purposes as well asfor use in conservation strategy


1964 ◽  
Vol 47 (3_Suppl) ◽  
pp. S28-S36
Author(s):  
Kailash N. Agarwal
Keyword(s):  

ABSTRACT Red cells were incubated in vitro with sulfhydryl inhibitors and Rhantibody with and without prior incubation with prednisolone-hemisuccinate. These erythrocytes were labelled with Cr51 and P32 and their disappearance in vivo after autotransfusion was measured. Prior incubation with prednisolone-hemisuccinate had no effect on the rate of red cell disappearance. The disappearance of the cells was shown to take place without appreciable intravascular destruction.


1960 ◽  
Vol XXXIV (II) ◽  
pp. 305-311 ◽  
Author(s):  
M. G. Woldring ◽  
A. Bakker ◽  
H. Doorenbos

ABSTRACT The red cell triiodothyronine uptake technique as used in our hospital is described. Incubation time is of almost no importance. The temperature during incubation should be 37° C. Further improvement of the technique is obtained when all blood samples are brought up to 40 % haematocrit prior to incubation. Clinical results are discussed. It is yet too early to give a definite assessment of its clinical value, but it is definitely superior to the measurement of the BMR.


2019 ◽  
Vol 26 (7) ◽  
pp. 512-522
Author(s):  
Xian Li ◽  
Long Xia ◽  
Xiaohui Ouyang ◽  
Qimuge Suyila ◽  
Liya Su ◽  
...  

<P>Background: Despite new agent development and short-term benefits in patients with Colorectal Cancer (CRC), metastatic CRC cure rates have not improved due to high rates of oxaliplatin resistance and toxicity. There is an urgent need for effective tools to prevent and treat CRC and reduce morbidity and mortality of CRC patients. Exploring the effects of bioactive peptides on the antitumor to CRC was of vital importance to the clinical application. </P><P> Objective: This study aimed to investigate the therapeutic impact of Anticancer Bioactive Peptides (ACBP) on anticancer effect of oxaliplatin (LOHP) in human colorectal cancer xenografts models in nude mice. </P><P> Methods: HCT-116 cells were cultured in vitro via CCK-8 assays and the absorbance was measured at 450 nm. Apoptosis and cell cycle were assessed by Flow Cytometry (FCM) in vitro. HCT-116 human colorectal cancer cells inoculated subcutaneously in nude mice of treatment with PBS (GG), ACBP, LOHP, ACBP+LOHP (A+L) in vivo. The quality of life was assessed by dietary amount of nude mice, the weight of nude mice, inhibition rates, tumor weight and tumor volume. Immunohistochemistry and RT-qPCR method was conducted to determine the levels of apoptosisregulating proteins/genes in transplanted tumors. </P><P> Results: ACBP induced substantial reductions in viable cell numbers and apoptosis of HCT116 cells in combined with LOHP in vitro. Compared with the control GG group, ACBP combined low dose oxaliplatin (U) group demonstrated significantly different tumor volume, the rate of apoptosis, the expression levels of Cyt-C, caspase-3,8,9 proteins and corresponding RNAs (P<0.05). The expression of pro-apoptotic proteins in the cytoplasm around the nucleus was significantly enhanced by ACBP. Short term intermittent use of ACBP alone indicted a certain inhibitory effect on tumor growth, and improve the quality of life of tumor bearing nude mice. </P><P> Conclusion: ACBP significantly increased the anti-cancer responses of low dose oxaliplatin (L-LOHP), thus, significantly improving the quality of life of tumor-bearing nude mice.</P>


Cancers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2428
Author(s):  
Frank Liang ◽  
Azar Rezapour ◽  
Peter Falk ◽  
Eva Angenete ◽  
Ulf Yrlid

TILs comprise functionally distinct conventional and unconventional T cell subsets and their role in responses to CRC treatments is poorly understood. We explored recovery of viable TILs from cryopreserved tumor biopsies of (chemo)-radiated patients with rectal cancer to establish a platform for retrospective TIL analyses of frozen tumors from pre-selected study cohorts. Frequencies of TIL subsets and their capacity to mount IFN-γ responses in cell suspensions of fresh vs. cryopreserved portions of the same tumor biopsies were determined for platform validation. The percentages and proportions of CD4+ TILs and CD8+ cytotoxic T lymphocytes (CTLs) among total TILs were not affected by cryopreservation. While recovery of unconventional γδ T cells and mucosal-associated invariant T cells (MAIT cells) was stable after cryopreservation, the regulatory T cells (Tregs) were reduced, but in sufficient yields for quantification. IFN-γ production by in vitro-stimulated CD4+ TILs, CTLs, γδ T cells, and MAIT cells were proportionally similar in fresh and cryopreserved tumor portions, albeit the latter displayed lower levels. Thus, the proposed platform intended for TIL analyses on cryopreserved tumor biobank biopsies holds promises for studies linking the quantity and quality of TIL subsets with specific clinical outcome after CRC treatment.


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