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Canadian Journal of Anaesthesia =... Feb 2023Excess protamine contributes to coagulopathy following cardiopulmonary bypass (CPB) and may increase blood loss and transfusion requirements. The primary aim of this... (Observational Study)
Observational Study
PURPOSE
Excess protamine contributes to coagulopathy following cardiopulmonary bypass (CPB) and may increase blood loss and transfusion requirements. The primary aim of this study was to find the least amount of protamine necessary to neutralize residual heparin following CPB using the gold standard assays of anti-IIa and anti-Xa activity. Secondary objectives were to evaluate whether the post-CPB activated clotting time could be used as a surrogate marker for quantifying heparin neutralization.
METHODS
Twenty-eight consecutive patients undergoing elective cardiac surgery were enrolled. Protamine administration was standardized through an infusion pump at 25 mg·min. Blood samples were withdrawn prior to and following administration of 150, 200, 250, and 300 mg protamine and analyzed for activated clotting time and anti-IIa and -Xa activity.
RESULTS
Following a mean (standard deviation) cumulative heparin dose of 67,700 (19,400) units and a CPB duration of 113 (71) min, protamine requirements varied widely. Eight out of 25 (32%) patients showed complete neutralization of anti-IIa and -Xa activity at the first sampling point (150 mg protamine; protamine:heparin ratio, 0.3 [0.1]). A protamine:heparin ratio of 0.5 (0.2) was sufficient for heparin neutralization in > 90% of patients. After CPB, a low to mid-range activated clotting time correlated well with anti-IIa and -Xa activity.
CONCLUSIONS
The protamine:heparin ratio required to neutralize residual unfractionated heparin (UFH) following CPB is variable. A protamine:heparin ratio of 0.3 was sufficient to neutralize UFH in some patients, while a ratio of 0.5 is sufficient to neutralize both residual anti-IIa and -Xa activity in most patients. Larger studies are necessary to confirm these findings and evaluate their clinical implications.
STUDY REGISTRATION
ClinicalTrials.gov (NCT03787641); registered 26 December 2018.
Topics: Humans; Heparin; Protamines; Prospective Studies; Cardiac Surgical Procedures; Cohort Studies; Cardiopulmonary Bypass; Anticoagulants
PubMed: 36471142
DOI: 10.1007/s12630-022-02364-4 -
Human Reproduction (Oxford, England) Nov 2014How are protamine deficiencies associated with sperm head morphology in subfertile men?
STUDY QUESTION
How are protamine deficiencies associated with sperm head morphology in subfertile men?
SUMMARY ANSWER
The prevalence of morphological variations and large nuclear vacuoles was slightly higher in protamine-deficient spermatozoa than in non-deficient spermatozoa.
WHAT IS KNOWN ALREADY
A protamine deficiency was previously reported to be associated with an abnormal sperm morphology; however, how they are related to each other remains unclear. This is further confounded by a number of protamine-deficient spermatozoa having a normal head morphology.
STUDY DESIGN, SIZE, DURATION
This is a cross-sectional study, including 36 men diagnosed with male factor infertility or participating in an assisted reproduction program. To assess sperm head morphology, this study analyzed 2400 spermatozoa with a protamine deficiency and 2400 spermatozoa with a normal protamine status. An additional 21 men were analyzed to examine DNA fragmentation and its relationship with protamine deficiencies and sperm head morphologies.
PARTICIPANTS/MATERIALS, SETTING, METHODS
The morphology of the sperm head was evaluated based on its shape, size and nuclear vacuoles at a magnification of >6000×. Using elliptic Fourier analysis, the shape was summarized into four numeric variables. The protamine status was evaluated with chromomycin A3 (CMA3). Sperm head size, vacuoles and shape were compared between protamine-deficient and non-deficient spermatozoa. DNA fragmentation was evaluated with the terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) assay. The percentages of protamine-deficient spermatozoa and DNA fragmentation were compared between spermatozoa with morphologically normal heads and those with abnormal heads.
MAIN RESULTS AND THE ROLE OF CHANCE
Variations in head size (P < 0.0001) and shape (P < 0.0001) were significantly higher, with narrower (P < 0.001), more fan-shaped (P < 0.01) and more square-shaped forms (P < 0.001) in protamine-deficient spermatozoa than in non-deficient spermatozoa; however, the distribution of morphological variations markedly overlapped. Protamine deficiencies were more frequently observed in spermatozoa with large nuclear vacuoles than in those without them (32.0 ± 3.1 versus 39.4 ± 2.9%, P < 0.001). The percentage of protamine-deficient spermatozoa was significantly lower in spermatozoa with a normal head morphology than in those with an abnormal head morphology (25.4 ± 2.6 versus 38.0 ± 2.5%, P < 0.001). The percentage of DNA fragmentation was significantly higher in protamine-deficient spermatozoa than in non-deficient spermatozoa (11.3 ± 2.1 versus 1.6 ± 0.6%, P < 0.001), and was lower in spermatozoa with a normal head morphology than in those with an abnormal head morphology (2.6 ± 0.7 versus 6.4 ± 0.2%, P < 0.001).
LIMITATIONS, REASONS FOR CAUTION
We were unable to discriminate the kind of protamines or quantify the extent of the protamine deficiency in spermatozoa using the CMA3 staining method.
WIDER IMPLICATIONS OF THE FINDINGS
This study provided a novel insight into how abnormal protamination affects sperm head morphology as well as the relationship between sperm head morphology and its own molecular integrity. Our results will contribute to a deeper understanding of the benefits and limitations of the morphological selection of spermatozoa for ICSI.
STUDY FUNDING/COMPETING INTERESTS
This study was supported by a JSPS Grant-in-Aid for the Encouragement of Scientists (25931009, 26931010). All authors have no conflicts of interest to disclose.
TRIAL REGISTRATION NUMBER
N/A.
Topics: Adult; Cell Shape; Cross-Sectional Studies; DNA Fragmentation; Humans; Infertility, Male; Male; Middle Aged; Protamines; Sperm Head; Spermatozoa; Vacuoles
PubMed: 25190724
DOI: 10.1093/humrep/deu225 -
PLoS Genetics Jun 2022Protamines are unique sperm-specific proteins that package and protect paternal chromatin until fertilization. A subset of mammalian species expresses two protamines...
Protamines are unique sperm-specific proteins that package and protect paternal chromatin until fertilization. A subset of mammalian species expresses two protamines (PRM1 and PRM2), while in others PRM1 is sufficient for sperm chromatin packaging. Alterations of the species-specific ratio between PRM1 and PRM2 are associated with infertility. Unlike PRM1, PRM2 is generated as a precursor protein consisting of a highly conserved N-terminal domain, termed cleaved PRM2 (cP2), which is consecutively trimmed off during chromatin condensation. The carboxyterminal part, called mature PRM2 (mP2), interacts with DNA and together with PRM1, mediates chromatin-hypercondensation. The removal of the cP2 domain is believed to be imperative for proper chromatin condensation, yet, the role of cP2 is not yet understood. We generated mice lacking the cP2 domain while the mP2 is still expressed. We show that the cP2 domain is indispensable for complete sperm chromatin protamination and male mouse fertility. cP2 deficient sperm show incomplete protamine incorporation and a severely altered protamine ratio, retention of transition proteins and aberrant retention of the testis specific histone variant H2A.L.2. During epididymal transit, cP2 deficient sperm seem to undergo ROS mediated degradation leading to complete DNA fragmentation. The cP2 domain therefore seems to be a key aspect in the complex crosstalk between histones, transition proteins and protamines during sperm chromatin condensation. Overall, we present the first step towards understanding the role of the cP2 domain in paternal chromatin packaging and open up avenues for further research.
Topics: Animals; Chromatin; Histones; Humans; Infertility, Male; Male; Mammals; Mice; Protamines; Semen; Spermatozoa
PubMed: 35763544
DOI: 10.1371/journal.pgen.1010272 -
Anticancer Research Dec 2018Anticoagulation therapy is often used to prevent stroke in patients with thyroid cancer. However, the effects of heparin and protamine on thyroid cancer are unknown;...
BACKGROUND
Anticoagulation therapy is often used to prevent stroke in patients with thyroid cancer. However, the effects of heparin and protamine on thyroid cancer are unknown; therefore, we examined the response of thyroid cancer cell lines to heparin and protamine.
MATERIALS AND METHODS
Cytotoxic assay for heparin-protamine treatment was examined on SW1736, 8505c and 8305c cell lines.
RESULTS
The half-maximal inhibitory concentration of the heparin-protamine treatment was 82.1 μg/ml for the SW1736 cell line, 10.4 μg/ml for 8505c, and 17.8 μg/ml for 8305c. Each cell line expresses fibronectin, with SW1736 expressing mutant fibronectin; thus, it is possible that mutant fibronectin may prevent the antitumor effect in SW1736 cells.
CONCLUSION
The SW1736 cell line did not show any antitumor effect after heparin-protamine treatment. Further research is needed on why heparin-protamine treatment does not exert an antitumor effect on SW1736 cells, and the results of this research could mean that the heparin-protamine treatment might be used to provide antitumor effects in some thyroid cancer cases.
Topics: Anticoagulants; Antineoplastic Agents; Cell Line, Tumor; Fibronectins; Gene Expression Regulation, Neoplastic; Heparin; Humans; Protamines; Thyroid Neoplasms
PubMed: 30504387
DOI: 10.21873/anticanres.13046 -
Biology of Reproduction Nov 2021While an E3 ubiquitin ligase, RNF8, was initially reported to be required for histone-to-protamine exchange in spermiogenesis, we subsequently demonstrated that RNF8 is...
While an E3 ubiquitin ligase, RNF8, was initially reported to be required for histone-to-protamine exchange in spermiogenesis, we subsequently demonstrated that RNF8 is not involved in this process. Nevertheless, reflecting a lingering misunderstanding in the field, a growing number of studies have continued to postulate a requirement for RNF8 in the histone-to-protamine exchange. For example, a recent study claimed that a mouse PIWI protein, MIWI, controls RNF8-mediated histone-to-protamine exchange. Here, confirming our earlier conclusions, we show that RNF8 is required neither for the establishment of histone H4K16 acetylation, which is an initial step in histone removal during spermiogenesis, nor for the incorporation of two protamine proteins, PRM1 and PRM2. Thus, whereas RNF8 mediates ubiquitination of H2A on the sex chromosomes in meiosis, during the prior stage of spermatogenesis, our genetic evidence underscores that RNF8 is not involved in histone-to-protamine exchange.
Topics: Acetylation; Animals; Biological Transport; Chromatin Assembly and Disassembly; Histones; Mice; Mice, Knockout; Protamines; Sex Chromosomes; Spermatogenesis; Ubiquitin-Protein Ligases; Ubiquitination
PubMed: 34225362
DOI: 10.1093/biolre/ioab132 -
Methods in Molecular Biology (Clifton,... 2017Nanoparticles of defined size can be easily obtained by simply mixing Protamine, a pharmaceutical drug that is used to neutralize heparin after surgery, and RNA in the...
Nanoparticles of defined size can be easily obtained by simply mixing Protamine, a pharmaceutical drug that is used to neutralize heparin after surgery, and RNA in the form of oligonucleotides or messenger RNA. Depending on the concentrations of the two reagents and their salt contents, homogenous nanoparticles with a mean diameter of 50 to more than 1000 nm can spontaneously be generated. RNA is a danger signal because it is an agonist of for example TLR-3, -7, and -8; therefore, Protamine-RNA nanoparticles are immunostimulating. We and others have shown in vitro that nanoparticle size and interferon-alpha production by human peripheral blood mononuclear cells (PBMCs) are inversely correlated. Conversely, nanoparticle size and TNF-alpha production by PBMCs are positively correlated (Rettig et al., Blood 115:4533-4541, 2010). Particles of less than 450 nm are most frequently used for research and clinical applications because they are very stable, remain polydispersed and induce interferon-alpha proteins, which are a natural antiviral and anticancer protein family with 12 members in humans. Herein, we describe a method to generate 130 nm nanoparticles as well as some of their physical and biological characteristics.
Topics: Heparin; Humans; Immunization; Interferon-alpha; Leukocytes, Mononuclear; Nanoparticles; Particle Size; Protamines; RNA, Messenger
PubMed: 27987148
DOI: 10.1007/978-1-4939-6481-9_9 -
Scandinavian Cardiovascular Journal :... 2016Platelet dysfunction is an important cause of postoperative bleeding after cardiac surgery. Protamine is routinely used for reversal of heparin after cardiopulmonary...
OBJECTIVES
Platelet dysfunction is an important cause of postoperative bleeding after cardiac surgery. Protamine is routinely used for reversal of heparin after cardiopulmonary bypass (CBP), but may affect platelet aggregation. We assessed changes in platelet function in relation to protamine administration.
DESIGN
Platelet aggregation was analyzed by impedance aggregometry before and after protamine administration in 25 adult cardiac surgery patients. Aggregation was also studied after in vitro addition of heparin and protamine. The activators adenosine diphosphate (ADP), thrombin receptor activating peptide-6 (TRAP), arachidonic acid (AA) and collagen (COL) were used.
RESULTS
Platelet aggregation was reduced by approximately 50% after in vivo protamine administration; ADP 640 ± 230 (AU*min, mean ± SD) to 250 ± 160, TRAP 939 ± 293 to 472 ± 260, AA 307 ± 238 to 159 ± 143 and COL 1022 ± 350 to 506 ± 238 (all p < 0.001). Aggregation was also reduced after in vitro addition of protamine alone with activators ADP from 518 ± 173 to 384 ± 157 AU*min p < 0.001, and AA 449 ± 311 to 340 ± 285 (p < 0.01) and protamine combined with heparin (1:1 ratio) with activators ADP to 349 ± 160 and AA to 308 ± 260 (both p < 0.001); and COL from 586 ± 180 to 455 ± 172 (p < 0.05).
CONCLUSIONS
Protamine given after CPB markedly reduces platelet aggregation. Protamine added in vitro also reduces platelet aggregation, by itself or in combination with heparin.
Topics: Aged; Anticoagulants; Cardiac Surgical Procedures; Cardiopulmonary Bypass; Down-Regulation; Female; Heparin; Heparin Antagonists; Humans; Male; Middle Aged; Pilot Projects; Platelet Aggregation; Platelet Function Tests; Postoperative Hemorrhage; Protamines; Risk Factors; Treatment Outcome
PubMed: 26402229
DOI: 10.3109/14017431.2015.1099720 -
ACS Applied Bio Materials Jul 2023Molybdenum-based nanomaterials with variable oxidation states can be developed as nanozyme catalysts. In this work, we developed a one-pot method for the preparation of...
Molybdenum-based nanomaterials with variable oxidation states can be developed as nanozyme catalysts. In this work, we developed a one-pot method for the preparation of molybdenum disulfide assisted by protein. Protamine was used as a cationic template to link molybdate anions and form complexes. During hydrothermal synthesis, protamine can affect the nucleation process of molybdenum disulfide and inhibit their aggregation, which facilitates the fabrication of small-sized molybdenum disulfide nanoparticles. Moreover, the abundant amino/guanidyl groups of protamine could both physically adsorb and chemically bond to molybdenum disulfide and further modulate the crystal structures. The optimized size and crystalline structure enabled a higher exposure of active sites, which enhanced the peroxidase-like activity of molybdenum disulfide/protamine nanocomposites. Meanwhile, the antibacterial activity of protamine was retained in the molybdenum disulfide/protamine nanocomposites, which could synergize with the peroxidase-like activity of molybdenum disulfide to kill bacteria. Therefore, the molybdenum disulfide/protamine nanocomposites are good candidates for antibacterial agents with lower chances of antimicrobial resistance. This study establishes an easy way to design artificial nanozymes by compounding suitable components.
Topics: Molybdenum; Biomimetics; Nanocomposites; Protamines; Peroxidases; Anti-Bacterial Agents
PubMed: 37317061
DOI: 10.1021/acsabm.3c00341 -
Biomaterials Science Apr 2022Designing and building artificial nanodevices and nanoarchitectures in living systems are extremely intriguing subjects in nanotechnology and synthetic biology. Taking...
Designing and building artificial nanodevices and nanoarchitectures in living systems are extremely intriguing subjects in nanotechnology and synthetic biology. Taking advantage of cellular machinery and endogenous biomacromolecules, such as proteins, is key to achieving the precise and sophisticated manipulation of nanoarchitectures. In this study, we proposed a protein-mediated DNA self-assembly strategy in a molecular crowding environment. By carefully controlling the surface charge of basic nuclear proteins in a crowding environment that mimicked the intracellular environment, we demonstrated that highly positively charged protamine can assemble individual DNA strands into defined structures similar to a catalytic process manner. Successful self-assembly required an optimized protamine surface charge and a crowding environment; otherwise, this self-assembly was impossible. Polyacrylamide gel electrophoresis (PAGE), atomic force microscopy (AFM), and dynamic light scattering (DLS) results showed that tile-based DNA tubular structures, tetrahedra, and two dimensional DNA origami structures were successfully assembled. We inferred that the assembly process occurred between the arginine-rich domain of protamine and DNA strands that repeatedly interacted with each other in the viscous system. The current study provides a potential strategy to construct nanodevices in living systems and presents an alternative protein-DNA interaction for the potential fabrication of protein-DNA hybrid nanomaterials.
Topics: DNA; Humans; Microscopy, Atomic Force; Nanostructures; Nanotechnology; Nucleic Acid Conformation; Protamines
PubMed: 35289345
DOI: 10.1039/d1bm02017j -
European Journal of Pharmaceutical... Jul 2015Application of oligonucleotides as active compounds has become a crucial field of pharmaceutical research in recent years. In order to improve inadequate transfection... (Review)
Review
Application of oligonucleotides as active compounds has become a crucial field of pharmaceutical research in recent years. In order to improve inadequate transfection rate and to avoid rapid enzymatic degradation of antisense oligonucleotides (AS-ODNs) a novel nanoparticulate delivery system was reported by our group at the beginning of 2000. AS-ODNs are condensed by the polycationic peptide protamine into solid particles in the size range of 100-200nm. Nanoparticle formation is driven by a self-assembling process based on electrostatic interactions between the oppositely charged biomolecules. This new delivery system was named "proticles" and showed very efficient protection against enzymatic digestion, high transfection rates and significant antisense effects in vitro. Throughout broader research, this promising approach was enlarged, and AS-ODNs were replaced by siRNA or CpG-oligonucleotides to address the aspect of immune-modulation and vaccination. More recent studies on proticles verified upscaling of the self-assembling process as well as the potential of proticle formulations for active drug targeting, like tumor- or atherosclerotic plaque targeting. Thereby also the application for diagnostic purposes was emphasized. This review will focus on the characterization of the nucleoprotein protamine as well as on the variety of possible nucleotides/peptides which were already assembled into the proticle matrix. Furthermore it will provide an insight into the broad area of application where proticles can present a valuable tool for successful oligonucleotide delivery.
Topics: Animals; Drug Delivery Systems; Humans; Immunologic Factors; Nanoparticles; Oligonucleotides; Protamines
PubMed: 25896372
DOI: 10.1016/j.ejps.2015.04.009