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The Journal of Small Animal Practice Sep 2022Smarter understanding of diabetes pathophysiology and pharmacology of insulin therapy can lead to better clinical outcomes. Rather than looking for an insulin... (Review)
Review
Smarter understanding of diabetes pathophysiology and pharmacology of insulin therapy can lead to better clinical outcomes. Rather than looking for an insulin formulation that is considered "best" for a general population, it could be appropriate to seek the "smart" insulin choice, tailored to the specific clinical situation. Different treatment goals should be considered, with pros and cons to each. Ideally, insulin therapy in most diabetic dogs should mimic a "basal-bolus" pattern. The "intermediate"-acting insulin formulations might provide better "bolus" treatment in dogs than the rapid-acting formulations used in people. In patients with some residual beta cell function such as many diabetic cats, administering only a "basal" insulin might lead to complete normalisation of blood glucose concentrations. Insulin suspensions (neutral protamine Hagedorn, neutral protamine Hagedorn/regular mixes, lente and protamine zinc insulin) as well as insulin glargine U100 and detemir are "intermediate"-acting formulations that are administered twice daily. For a formulation to be an effective and safe "basal" insulin, its action should be roughly the same every hour of the day. Currently, only insulin glargine U300 and insulin degludec meet this standard in dogs, whereas in cats, insulin glargine U300 is the closest option.
Topics: Animals; Blood Glucose; Cat Diseases; Cats; Diabetes Mellitus; Dog Diseases; Dogs; Humans; Hypoglycemic Agents; Insulin; Insulin Glargine; Protamines
PubMed: 35560042
DOI: 10.1111/jsap.13507 -
Protein and Peptide Letters 2018Protamines are the most abundant sperm nuclear proteins and pack approximately the 92-98% of the mammalian sperm DNA. In mammals, two types of protamines have been... (Review)
Review
BACKGROUND
Protamines are the most abundant sperm nuclear proteins and pack approximately the 92-98% of the mammalian sperm DNA. In mammals, two types of protamines have been described, the Protamine 1 (P1) and the Protamine 2 (P2) family. The deregulation of the relative P1/P2 ratio has been correlated to DNA damage, alterations in seminal parameters, and low success rate of assisted reproduction techniques. Additionally, the extraction and analysis of protamines have been important to understand the fundamental aspects of the sperm chromatin structure and function, protamine sequence conservation among species, and sperm chromatin alterations present in infertile males. However, protamines show a particular chemical nature due to its special amino acid sequence, extremely rich in arginine and cysteine residues. Because of these peculiar characteristics of protamines, their extraction and analysis is not as straightforward as the analysis of other chromatin-associated proteins, for which many detailed protocols are already available.
CONCLUSION
A step-by-step protocol was needed to facilitate protamine analysis to researchers interested in their implementation. Therefore, in order to contribute to fulfill this need, here we provide a detailed protocol, which should be useful to research teams and laboratories interested in the protamine field. In addition, we also briefly review the different studies published so far on protamine alterations and male infertility.
Topics: Animals; Humans; Male; Protamines; Spermatozoa
PubMed: 29651936
DOI: 10.2174/0929866525666180412155205 -
ACS Nano Aug 2021Packaging paternal genome into tiny sperm nuclei during spermatogenesis requires 10-fold compaction of DNA, corresponding to a 10-20 times higher compaction than in...
Packaging paternal genome into tiny sperm nuclei during spermatogenesis requires 10-fold compaction of DNA, corresponding to a 10-20 times higher compaction than in somatic cells. While such a high level of compaction involves protamine, a small arginine-rich basic protein, the precise mechanism at play is still unclear. Effective pair potential calculations and large-scale molecular dynamics simulations using a simple idealized model incorporating solely electrostatic and steric interactions clearly demonstrate a reversible control on DNA condensates formation by varying the protamine-to-DNA ratio. Microscopic states and condensate structures occurring in semidilute solutions of short DNA fragments are in good agreement with experimental phase diagram and cryoTEM observations. The reversible microscopic mechanisms induced by protamination modulation should provide valuable information to improve a mechanistic understanding of early and intermediate stages of spermatogenesis where an interplay between condensation and liquid-liquid phase separation triggered by protamine expression and post-translational regulation might occur. Moreover, recent vaccines to prevent virus infections and cancers using protamine as a packaging and depackaging agent might be fine-tuned for improved efficiency using a protamination control.
Topics: Male; Humans; Protamines; Spermatozoa; Semen; DNA Packaging; DNA
PubMed: 34328301
DOI: 10.1021/acsnano.1c02337 -
Asian Cardiovascular & Thoracic Annals Jan 2021Heparin is used for anticoagulation during cardiopulmonary bypass. After weaning from bypass, protamine is administered to neutralize the effects of heparin and thus...
BACKGROUND
Heparin is used for anticoagulation during cardiopulmonary bypass. After weaning from bypass, protamine is administered to neutralize the effects of heparin and thus reestablish hemostasis. Rotational thrombelastometry has been shown to discriminate between heparin and other impairing effects on coagulation. We analyzed the interaction of heparin and protamine under different conditions of overdosage in an in-vitro trial.
METHODS
Blood samples were taken from 17 healthy volunteers, separated, and spiked in vitro with heparin, protamine for heparin neutralization, an overdosage of protamine, and two dosages of re-heparinization to evaluate heparin effects under the condition of protamine overdosage. All samples were analyzed in a standard ROTEM rotational thromboelastometry device after intrinsic activation with and without addition of heparinase. Coagulation time, maximum clot firmness, and clot formation time were recorded.
RESULTS
Heparin led to prolongation of coagulation and clot formation times in the test without heparinase. Adequate protamine addition normalized the test, and overdosage of protamine led to significant prolongation of both times. Addition of heparin in the presence of protamine overdosage normalized these parameters.
CONCLUSION
We reconfirmed that the ROTEM device enables discrimination of the effects heparin and protamine on coagulation and detection of the coagulation-impairing effects of protamine overdosage. Furthermore, we were able to show a positive effect on coagulation times by heparin in the presence of protamine overdosage. Because this was an in-vitro study, these findings need to be confirmed in vivo, requiring further research.
Topics: Adult; Anticoagulants; Blood Coagulation; Drug Overdose; Female; Heparin; Heparin Antagonists; Humans; Male; Protamines; Thrombelastography; Time Factors
PubMed: 32854516
DOI: 10.1177/0218492320955065 -
Current Opinion in Cell Biology Apr 2022The sperm genome is tightly packed into a minimal volume of sperm nuclei. Sperm chromatin is highly condensed by protamines (PRMs) after histone-protamine replacement,... (Review)
Review
The sperm genome is tightly packed into a minimal volume of sperm nuclei. Sperm chromatin is highly condensed by protamines (PRMs) after histone-protamine replacement, and the majority of the sperm genome forms a nucleo-protamine structure, namely, the PRM-DNA complex. The outline of sperm chromatin structure was proposed 30 years ago, and the details have been explored by approaches from several independent research fields including male reproduction and infertility, DNA biopolymer, and most recently, genome-wide sequence-based approaches. In this review, the history of research on sperm chromatin structure is briefly described, and the progress of recent related studies is summarized to obtain a more integrated view for the sperm chromatin, an extremely compacted "black box."
Topics: Chromatin; Chromatin Assembly and Disassembly; DNA; Humans; Male; Protamines; Spermatozoa
PubMed: 35344802
DOI: 10.1016/j.ceb.2022.102075 -
Biochemical and Biophysical Research... Apr 2021Studying thermal stability of proteins not only provides insight into protein structure but also is instrumental in identifying previously unknown interaction partners....
Studying thermal stability of proteins not only provides insight into protein structure but also is instrumental in identifying previously unknown interaction partners. We develop a machine learning strategy that combines orthogonal partial least squares regression and stability screening of Silver Bullets Bio library to identify biologically active molecules that enhance protein stability. This strategy proves effective in extracting the stability-enhancing molecules for SMYD5, a histone lysine methyltransferase that regulates chromosome integrity. Protamine, a histone substitute in chromatin condensation during spermatogenesis, is identified as the most influential molecule to enhance SMYD5 thermal stability. We find that the C-terminal poly-glutamic acid tract (poly-E) and a 30-residue insertion in MYND domain (M-insertion), which are unique to SMYD5, regulate the structural stability. However, protamine plays a dominant role in SMYD5 stability, and in the presence of protamine, the poly-E tract or M-insertion loses its ability to affect the stability. The stability-enhancing effect of protamine is SMYD5 specific, and for SMYD2, a closely related homolog, protamine exhibits opposite, destabilizing effects. We find that both SMYD5 and SMYD2 interact with protamine, where SMYD5 interaction is independent of the poly-E tract and M-insertion. Protamine not only helps provide insight into the structure-stability relationships of SMYD5, but also suggests a potential functional link of SMYD5 to spermatogenesis. SMYD5 is a ubiquitously expressed gene with the highest expression in testis, especially in the seminiferous ducts that contain germ cells. Thus, our study opens up avenues that could help delineate major mechanisms underlying chromatin dynamics during spermatogenesis.
Topics: Chromatin; Chromatin Assembly and Disassembly; Humans; Machine Learning; Male; Methyltransferases; Models, Molecular; Protamines; Protein Binding; Protein Stability; Spermatozoa; Temperature
PubMed: 33676231
DOI: 10.1016/j.bbrc.2021.02.073 -
Journal of Materials Science. Materials... Apr 2022Calcium phosphates are key biomaterials in dental treatment and bone regeneration. Biomaterials must exhibit antibacterial properties to prevent microbial infection in...
Calcium phosphates are key biomaterials in dental treatment and bone regeneration. Biomaterials must exhibit antibacterial properties to prevent microbial infection in implantation frameworks. Previously, we developed various types of calcium phosphate powders (amorphous calcium phosphate, octacalcium phosphate (OCP), dicalcium phosphate anhydrate, and hydroxyapatite) with adsorbed protamine (which is a protein with antibacterial property) and confirmed their antibacterial property. In this study, as foundational research for the development of novel oral care materials, we synthesized calcium phosphate composite powders from three starting materials: i) OCP, which intercalates organic compounds, ii) protamine, which has antibacterial properties, and iii) F ion, which promotes the formation of apatite crystals. Through investigating the preparation concentration of the F ions and their loading into OCP, it was found that more F ion could be loaded at higher concentrations regardless of the loading method. It was also observed that the higher the preparation concentration, the more the OCP converted to fluorapatite. The synthesized calcium phosphate composite powders were evaluated for biocompatibility through proliferation of MG-63 cells, with none of the powders exhibiting any growth inhibition. Antimicrobial tests showed that the calcium phosphate composite powders synthesized with protamine and F ion by precipitation had enhanced antimicrobial properties than those synthesized by protamine adsorption. Thus, the calcium phosphate composite powder prepared from OCP, protamine, and F ion forms the basis for promising antimicrobial biomaterials. Graphical abstract.
Topics: Anti-Bacterial Agents; Anti-Infective Agents; Calcium Phosphates; Fluorides; Powders; Protamines
PubMed: 35362837
DOI: 10.1007/s10856-022-06656-5 -
Nature Structural & Molecular Biology Aug 2023Conventional dogma presumes that protamine-mediated DNA compaction in sperm is achieved by electrostatic interactions between DNA and the arginine-rich core of...
Conventional dogma presumes that protamine-mediated DNA compaction in sperm is achieved by electrostatic interactions between DNA and the arginine-rich core of protamines. Phylogenetic analysis reveals several non-arginine residues conserved within, but not across species. The significance of these residues and their post-translational modifications are poorly understood. Here, we investigated the role of K49, a rodent-specific lysine residue in protamine 1 (P1) that is acetylated early in spermiogenesis and retained in sperm. In sperm, alanine substitution (P1(K49A)) decreases sperm motility and male fertility-defects that are not rescued by arginine substitution (P1(K49R)). In zygotes, P1(K49A) leads to premature male pronuclear decompaction, altered DNA replication, and embryonic arrest. In vitro, P1(K49A) decreases protamine-DNA binding and alters DNA compaction and decompaction kinetics. Hence, a single amino acid substitution outside the P1 arginine core is sufficient to profoundly alter protein function and developmental outcomes, suggesting that protamine non-arginine residues are essential for reproductive fitness.
Topics: Animals; Male; Mice; Amino Acids; Arginine; Chromatin; DNA; Genetic Fitness; Phylogeny; Protamines; Semen; Sperm Motility; Spermatozoa
PubMed: 37460896
DOI: 10.1038/s41594-023-01033-4 -
Molecules (Basel, Switzerland) Jan 2024Protamine is a cationic peptide derived from fish sperm and has several important functional properties: antibacterial properties, acting as a carrier for injectable...
Protamine is a cationic peptide derived from fish sperm and has several important functional properties: antibacterial properties, acting as a carrier for injectable insulin and as a heparin antagonist, combatting fatigue, etc. Thus, it has been widely used in medicinal applications and food products. Cultured is a type of pufferfish with a delicious taste that is popular in China, and its production is increasing significantly. Therefore, protamine was extracted via acid extraction from the sperm of and further isolated and purified via sephadex gel chromatography, ion exchange chromatography, and desalination chromatography. Furthermore, the physicochemical properties of protamine were investigated. The results showed that the sperm of the cultured were non-toxic, and the extracted and purified protamine had high contents of arginine (36.90%) and lysine (27.02%), respectively. The secondary structure of protamine was mainly β-folded and irregularly curled. Additionally, protamine exhibited high thermal stability with a denaturation temperature of 176 °C. This study would provide a theoretical basis for the structural analysis, bioactivity, and resource development of pufferfish protamine and help to promote the development of the pufferfish industry.
Topics: Male; Animals; Takifugu; Protamines; Semen; Heparin Antagonists; Anti-Bacterial Agents
PubMed: 38202846
DOI: 10.3390/molecules29010263 -
Catheterization and Cardiovascular... Jul 2023Access to the arterial circulation and full anticoagulation carries a risk of serious bleeding during and after percutaneous coronary intervention. Important sources of... (Review)
Review
Access to the arterial circulation and full anticoagulation carries a risk of serious bleeding during and after percutaneous coronary intervention. Important sources of bleeding include the arterial access site and coronary artery perforation. Prompt and effective management of hemorrhagic complications is an essential interventional skill. Protamine sulfate is well-known as a heparin reversal agent. Despite this, there is heterogeneity in the use of protamine during interventional procedures. While protamine is generally well-tolerated, it is associated with a risk of hypersensitivity reaction, including anaphylaxis, among others. The purpose of this review is to summarize the existing evidence about and experience with the use of protamine sulfate in the setting of percutaneous coronary and structural interventional procedures.
Topics: Humans; Protamines; Treatment Outcome; Hemorrhage; Blood Coagulation; Cardiac Catheterization
PubMed: 37172213
DOI: 10.1002/ccd.30679