Hemolysis induced by a Bellofram blood pump; chronic in vitro evaluation

In the application of extracorporeal pumping systems to chronic assisted circulation, hemolysis may be an important factor. A reciprocating pump utilizing rolling diaphragms has been tested for production of free plasma hemoglobin. In a recirculating system, 1,500 ml of freshly drawn heparinized dog blood was circulated at a flow rate of 3,000 ml/min against pressures of 100, 200, and 300 mm Hg for from 1 to 24 hr. Hemolysis was determined by a modified benzidine method. Recirculation for 1 hr against a height of 142 cm produced 3.6 mg/100 ml, 1.8 mg/100 ml, and 2.4 mg/100 ml free plasma hemoglobin, respectively, for hemolysis indices of 0.001- 0.0013. When the same priming volume was recirculated at 3,000 ml/min and 300 mm Hg pressure for 24 hr, the average hourly rate of free hemoglobin accumulation was 0.5 mg/100 ml, an index of 0.0006. These rates of hemolysis are comparable to the estimated normal human production of free plasma hemoglobin. The extremely low rate of hemolysis produced by this pump is related to the noncompressive action of the rolling diaphragms and flap valves. The characteristics of its performance make this pump ideally suited to chronic perfusion. extracorporeal pump; assisted circulation; rolling diaphragm; reciprocating pump Submitted on June 18, 1964

Download Full-text

CORRELATION OF FREE HEMOGLOBIN LEVEL AND PLASMA NITRIC OXIDE IN PACKED RED CELL DURING BLOOD BANK STORAGE PERIOD

INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY ◽

10.24293/ijcpml.v25i1.1485 ◽

2019 ◽

Vol 25 (1) ◽

pp. 26

Author(s):

Ricca Fitria ◽

Zelly Dia Rofinda ◽

Desywar Desywar

Keyword(s):

Nitric Oxide ◽

Storage Period ◽

Blood Bank ◽

Hemoglobin Level ◽

Free Hemoglobin ◽

Cross Sectional ◽

Plasma Hemoglobin ◽

Nitric Oxide Level ◽

Spearman Test ◽

Free Plasma

Stored red blood cells undergo morphological and biochemical changes with increased storage time, commonly refferred as the storage lesion, reduced integrity of erythrocyte membrane, causing hemolysis and increased free plasma hemoglobin level. Nitric Oxide (NO) is an endogenous vasodilator with the major role in vascular and blood flow regulation. Accumulation of free hemoglobin (fHb) during storage RBC hemolysis is thought to be correlated with elevated NO consuming causes low NO level that significantly impair endothelial function. This study aim to was to analyze the correlation of free Hb and plasma nitric oxide level during PRC storage period in the blood bank. This research was an analytic study with a cross-sectional design to 14 unit PRC those were stored in Dr. M. Djamil Padang Hospital Blood Bank. The study was conducted from May 2016 to August 2017. The assay was performed for 28 days of storage with one week intervals. Free plasma hemoglobin and NO level were analyzed by cyanmethemoglobin and colorimetry method respectively. Spearman test was used to access the correlation between free Hb level and nitric oxide plasma with p<0.05 means significant. Most donors were male (85.7%) with age range were 33(9) years. Free Hb and nitric oxide level were significantly differenced in each week storage periods (p<0.05). Two parameters was significantly had moderate negative correlation during storage periods (r=-0.56; p=0.01).

Download Full-text

Presence and Possible Origin of ɛ(γ-Glutamyl)Lysine Isodipeptide in Human Plasma

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648380 ◽

1992 ◽

Vol 67 (01) ◽

pp. 060-062 ◽

Cited By ~ 9

Author(s):

J Harsfalvi ◽

E Tarcsa ◽

M Udvardy ◽

G Zajka ◽

T Szarvas ◽

...

Keyword(s):

Human Plasma ◽

Fibrinolytic Therapy ◽

Normal Human Plasma ◽

Normal Human ◽

Hplc Technique ◽

Significant Change

Summaryɛ(γ-glutamyl)lysine isodipeptide has been detected in normal human plasma by a sensitive HPLC technique in a concentration of 1.9-3.6 μmol/1. Incubation of in vitro clotted plasma at 37° C for 12 h resulted in an increased amount of isodipeptide, and there was no further significant change when streptokinase was also present. Increased in vivo isodipeptide concentrations were also observed in hypercoagulable states and during fibrinolytic therapy.

Download Full-text

Inhibition of the Activities of α2-Plasmin inhibitor (PI) and CI inhibitor (CI-Inh) by the Synthetic Fibrinolytic Agent, 3-Hydroxypropyl Flufenamide (Flu-HPA)

10.1055/s-0038-1665785 ◽

1979 ◽

Author(s):

L Miles ◽

J Burnier ◽

M Verlander ◽

M Goodman ◽

A Kleiss ◽

...

Keyword(s):

Plasminogen Activators ◽

Inhibitory Effect ◽

Mechanisms Of Action ◽

Flufenamic Acid ◽

Clot Lysis ◽

Factor Xiia ◽

Dependent Inhibition ◽

Normal Human ◽

Plasmin Inhibitor

Flu-HPA is one of a series of flufenamic acid derivations that enhances plasminogen-dependent clot lysis in vitro. Studies of possible mechanisms of action of Flu-HPA were undertaken. The influence of Flu-HPA on the inhibition of purified plasmin by purified PI was studied. PI activity was assessed by its inhibition of the clevage of the tripeptide S-2251 (H-D-Val-Leu-Lys-pNA) by plasmin. Flu-HPA was dissolved in DMF or in methonol and preincubated with PI before addition of plasmin. At Flu-HPA concentrations greater than 1mM and up to 60mM, the inhibitory activity of PI was totally lost. The inhibitory effect of normal human plasma on plasmin was also completely abolished at concentrations of Flu-HPA between 2.5 and 40mM. The effect of Flu-HPA on the inhibition of purified plasma kallikrein by purified CI-Inh was also studied. CI-Inh activity was measured by its inhibition of cleavage of the tripeptide Bz-Pro-Phe-Arg-pNA by kallikrein. When Flu-HPA, dissolved in DMF or in methonol, was preincubated with CI-Inh, a concentration dependent inhibition of CI-Inh activity was observed. CI-Inh activity was abolished by concentrations of Flu-HPA greater than 1mM. Flu-HPA also inhibited the activity of CI-Inh on purified Factor XIIa. These observations suggest that this flufenamic acid derivative may enhance fibrinolysis not only by inhibiting PI activity but also by decreasing the inactivation of plasminogen activators by CI-Inh.

Download Full-text

The Effect of Practolol, In Vitro Generated Metabolites of Practolol and Chemical Analogues on the Rate of Growth of Human Skin Fibroblasts

Alternatives to Laboratory Animals ◽

10.1177/026119298401200205 ◽

1984 ◽

Vol 12 (2) ◽

pp. 89-97

Author(s):

Graham R. Elliott ◽

H.E. Amos ◽

James W. Bridges

Keyword(s):

Human Skin ◽

Psoriasis Patient ◽

Skin Fibroblasts ◽

Human Skin Fibroblasts ◽

Cell Type ◽

Normal Human Skin ◽

Rate Of Growth ◽

Structural Analogues ◽

Normal Human

The rate of growth of normal human skin fibroblasts was inhibited in a dose related, reversible, fashion by practolol (N-4-(2-hydroxy)-3 (1-methyl)-aminopropoxyphenylacetamine) (ID50 1.35 ± 0.14 x 10-3M), propranolol (1-(isopropylamino)-3(1-naphthyl-oxy)-2-propranolol) (ID50 0.145 ± 0.02 x 10-3M) and paracetamol (N-(4-hydroxyphenyl) acetamide) (ID50 0.85 ± 0.2 x 10-3M). Skin fibroblasts isolated from a psoriasis patient were more sensitive towards practolol (ID50 0.48 ± 0.14 x 10-3M) and propranolol (ID50 0.032 ± 0.002 x 10-3M), but less sensitive towards paracetamol (ID50 1.3 ± 0.07 x 10-3M). In vitro generated metabolites of practolol, using normal or Arochlor 1254-pretreated hamster liver preparations, and structural analogues of practolol had no effect upon the growth of either cell type.

Download Full-text

HGG-26. H3G34V MUTATION AFFECTS GENOMIC H3K36 METHYLATION IN PEDIATRIC GLIOMA

Neuro-Oncology ◽

10.1093/neuonc/noaa222.310 ◽

2020 ◽

Vol 22 (Supplement_3) ◽

pp. iii348-iii348

Author(s):

Tina Huang ◽

Andrea Piunti ◽

Elizabeth Bartom ◽

Jin Qi ◽

Rintaro Hashizume ◽

...

Keyword(s):

Western Blot ◽

Allelic Frequency ◽

Glioma Cell Line ◽

Fluorescent Imaging ◽

Wild Type ◽

Gene Expressions ◽

Pediatric Glioma ◽

Genes Encoding ◽

Normal Human

Abstract BACKGROUND Histone H3.3 mutation (H3F3A) occurs in 50% of cortical pediatric high-grade gliomas. This mutation replaces glycine 34 with arginine or valine (G34R/V), impairing SETD2 activity (H3K36-specific trimethyltransferase), resulting in reduced H3K36me on H3G34V nucleosomes relative to wild-type. This contributes to genomic instability and drives distinct gene expressions associated with tumorigenesis. However, it is not known if this differential H3K36me3 enrichment is due to H3G34V mutant protein alone. Therefore, we set to elucidate the effect of H3G34V on genomic H3K36me3 enrichment in vitro. METHODS Doxycycline-inducible short hairpin RNA (shRNA) against H3F3A was delivered via lentivirus to established H3G34V mutant pediatric glioma cell line KNS42, and H3G34V introduced into H3.3 wild type normal human astrocytes (NHA). Transfections were confirmed by western blot, fluorescent imaging, and flow cytometry, with resulting H3.3WT and H3K36me3 expression determined by western blot. H3.3WT, H3K36me3, and H3G34V ChIP-Seq was performed to evaluate genomic enrichment. RESULTS Complete knockdown of H3G34V was achieved with DOX-induced shRNA, with no change in total H3.3, suggesting disproportionate allelic frequency of genes encoding H3.3 (H3F3A and H3F3B). Modest increase in H3K36me3 occurred after H3F3A-knockdown from KNS42, suggesting H3G34V alone impacts observed H3K36me3 levels. Distinct H3K36me3 genomic enrichment was observed with H3G34V knock-in. CONCLUSIONS We demonstrate that DOX-inducible knockdown of H3F3A in an H3G34V mutant pediatric glioma cells and H3G34V mutation transduction in wild-type astrocytes affects H3K36me3 expression. Further evaluation by ChIP-Seq analysis for restoration of wild-type genomic H3K36me3 enrichment patterns with H3G34V knockdown, and mutant H3K36me3 patterns with H3G34V transduction, is currently underway.

Download Full-text

Scaffold Hopping of α-Rubromycin Enables Direct Access to FDA-Approved Cromoglicic Acid as a SARS-CoV-2 MPro Inhibitor

Pharmaceuticals ◽

10.3390/ph14060541 ◽

2021 ◽

Vol 14 (6) ◽

pp. 541

Author(s):

Hani A. Alhadrami ◽

Ahmed M. Sayed ◽

Heba Al-Khatabi ◽

Nabil A. Alhakamy ◽

Mostafa E. Rateb

Keyword(s):

Binding Free Energy ◽

Human Fibroblasts ◽

Critical Time ◽

Dynamics Simulation ◽

Direct Access ◽

Energy Calculation ◽

Wide Range ◽

Normal Human ◽

Novel Scaffold

The COVID-19 pandemic is still active around the globe despite the newly introduced vaccines. Hence, finding effective medications or repurposing available ones could offer great help during this serious situation. During our anti-COVID-19 investigation of microbial natural products (MNPs), we came across α-rubromycin, an antibiotic derived from Streptomyces collinus ATCC19743, which was able to suppress the catalytic activity (IC50 = 5.4 µM and Ki = 3.22 µM) of one of the viral key enzymes (i.e., MPro). However, it showed high cytotoxicity toward normal human fibroblasts (CC50 = 16.7 µM). To reduce the cytotoxicity of this microbial metabolite, we utilized a number of in silico tools (ensemble docking, molecular dynamics simulation, binding free energy calculation) to propose a novel scaffold having the main pharmacophoric features to inhibit MPro with better drug-like properties and reduced/minimal toxicity. Nevertheless, reaching this novel scaffold synthetically is a time-consuming process, particularly at this critical time. Instead, this scaffold was used as a template to explore similar molecules among the FDA-approved medications that share its main pharmacophoric features with the aid of pharmacophore-based virtual screening software. As a result, cromoglicic acid (aka cromolyn) was found to be the best hit, which, upon in vitro MPro testing, was 4.5 times more potent (IC50 = 1.1 µM and Ki = 0.68 µM) than α-rubromycin, with minimal cytotoxicity toward normal human fibroblasts (CC50 > 100 µM). This report highlights the potential of MNPs in providing unprecedented scaffolds with a wide range of therapeutic efficacy. It also revealed the importance of cheminformatics tools in speeding up the drug discovery process, which is extremely important in such a critical situation.

Download Full-text

Glucose Metabolism in Normal Human Thyroid Tissue in vitro

European Journal of Clinical Investigation ◽

10.1111/j.1365-2362.1972.tb00646.x ◽

1972 ◽

Vol 2 (4) ◽

pp. 213-219 ◽

Cited By ~ 13

Author(s):

J. Otten ◽

J. E. Dumont

Keyword(s):

Glucose Metabolism ◽

Thyroid Tissue ◽

Human Thyroid ◽

Human Thyroid Tissue ◽

Normal Human

Download Full-text

Anaerobic Glycolysis in Normal Human Erythrocytes Incubated in Vitro with Sodium Salicylate

Clinical Science ◽

10.1042/cs0490375 ◽

1975 ◽

Vol 49 (5) ◽

pp. 375-384

Author(s):

N. Worathumrong ◽

A. J. Grimes

Keyword(s):

Sodium Salicylate ◽

Lactate Production ◽

Glucose Consumption ◽

Human Erythrocytes ◽

Anaerobic Glycolysis ◽

Initial Stimulus ◽

Sodium Efflux ◽

Normal Human ◽

Concentration Changes

1. Some effects of sodium salicylate upon anaerobic glycolysis have been studied in normal human erythrocytes incubated for up to 6 h at 37°C in autologous sera. 2. Both glucose consumption and lactate production were stimulated by concentrations of salicylate up to 60 mmol/l but at the highest concentration used (90 mmol/l) an initial stimulus was followed by inhibition of glycolysis. 3. Losses occurred of adenosine 5′-triphosphate (ATP), adenosine 5′-diphosphate (ADP) and adenosine 5′-phosphate (AMP) at higher concentrations of salicylate and there was a concomitant increase of inorganic phosphate. 4. Other phosphate esters underwent concentration changes at higher concentrations of salicylate that reflected inadequate concentrations of ATP for glycolysis. 5. The rates of sodium efflux from, and potassium influx into, erythrocytes were unaffected by the presence of salicylate at concentrations sufficient to stimulate glycolysis.

Download Full-text

The anti-angiogenic isoforms of VEGF in health and disease

Biochemical Society Transactions ◽

10.1042/bst0371207 ◽

2009 ◽

Vol 37 (6) ◽

pp. 1207-1213 ◽

Cited By ~ 76

Author(s):

Yan Qiu ◽

Coralie Hoareau-Aveilla ◽

Sebastian Oltean ◽

Steven J. Harper ◽

David O. Bates

Keyword(s):

Splice Site ◽

Site Selection ◽

Age Related Macular Degeneration ◽

Splicing Factor ◽

Splice Site Selection ◽

Age Related ◽

Normal Human ◽

Radical Revision

Anti-angiogenic VEGF (vascular endothelial growth factor) isoforms, generated from differential splicing of exon 8, are widely expressed in normal human tissues but down-regulated in cancers and other pathologies associated with abnormal angiogenesis (cancer, diabetic retinopathy, retinal vein occlusion, the Denys–Drash syndrome and pre-eclampsia). Administration of recombinant VEGF165b inhibits ocular angiogenesis in mouse models of retinopathy and age-related macular degeneration, and colorectal carcinoma and metastatic melanoma. Splicing factors and their regulatory molecules alter splice site selection, such that cells can switch from the anti-angiogenic VEGFxxxb isoforms to the pro-angiogenic VEGFxxx isoforms, including SRp55 (serine/arginine protein 55), ASF/SF2 (alternative splicing factor/splicing factor 2) and SRPK (serine arginine domain protein kinase), and inhibitors of these molecules can inhibit angiogenesis in the eye, and splice site selection in cancer cells, opening up the possibility of using splicing factor inhibitors as novel anti-angiogenic therapeutics. Endogenous anti-angiogenic VEGFxxxb isoforms are cytoprotective for endothelial, epithelial and neuronal cells in vitro and in vivo, suggesting both an improved safety profile and an explanation for unpredicted anti-VEGF side effects. In summary, C-terminal distal splicing is a key component of VEGF biology, overlooked by the vast majority of publications in the field, and these findings require a radical revision of our understanding of VEGF biology in normal human physiology.

Download Full-text