-
Dental Materials Journal Oct 2010The DNA/protamine complex was prepared by a reaction between DNA and protamine sulfate solutions with stirring, and its cell viability, antibacterial effect and...
The DNA/protamine complex was prepared by a reaction between DNA and protamine sulfate solutions with stirring, and its cell viability, antibacterial effect and histopathological responses were examined. A water-insoluble white powder, DNA/protamine complex, with a porous structure was obtained. The molar binding ratio of the complex prepared from a solution containing equal amounts of DNA and protamine sulfate by weight was 0.038 and the efficiency of complex formation was 61%. In a cell culture test using MC-3T3-E1 mouse osteoblast cells, the complex showed less cytotoxicity than protamine sulfate alone and cell viabilities were more than 98%. A porous disk could be prepared easily and showed an antibacterial effect against Staphyrococcus aureus, Porphyromonas gingivalis and Prevotella intermedia in an antibacterial sensitivity test and a mild tissue response in vivo test. These results suggested that the DNA/protamine complex could be a useful biodegradable biomaterial with antibacterial effects.
Topics: 3T3 Cells; Animals; Anti-Bacterial Agents; Biocompatible Materials; Cell Survival; DNA; DNA Adducts; Implants, Experimental; Male; Materials Testing; Mice; Microbial Sensitivity Tests; Microscopy, Electron, Scanning; Osteoblasts; Porosity; Porphyromonas gingivalis; Prevotella intermedia; Protamines; Rats; Rats, Sprague-Dawley; Salmon; Solubility; Staphylococcus aureus; Subcutaneous Tissue; Surface Properties; Water
PubMed: 20733264
DOI: 10.4012/dmj.2009-131 -
Asian Journal of Andrology Mar 2010Semen samples collected from 28 male partners of infertile couples were divided into three equal aliquots and prepared with three selected media, such as PureSperm... (Comparative Study)
Comparative Study
Comparative study of the effects of three semen preparation media on semen analysis, DNA damage and protamine deficiency, and the correlation between DNA integrity and sperm parameters.
Semen samples collected from 28 male partners of infertile couples were divided into three equal aliquots and prepared with three selected media, such as PureSperm (Nidacon, Gothenburg, Sweden), Sil-Select Plus (Fertipro, Beernem, Belgium) and SpermGrad (Vitrolife, Gothenburg, Sweden). The differences in mean percentages of semen parameters were assessed by repeated measures analysis. Correlations of sperm DNA damage, as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and of protamine deficiency, as measured by chromomycin A3 (CMA3) staining with sperm parameters, were determined by Pearson's correlation. After preparation with all three media, sperm concentrations decreased (P < 0.05) while percentages of sperm with normal morphology increased (P < 0.05). Percentages of sperm motility, rapid motility and progressive motile concentration (PMC) increased (P < 0.05) for each of these parameters, PureSperm preparation gave the best results (P < 0.05). The percentage of DNA damage decreased in the PureSperm and Sil-Select Plus preparations (17.9% and 31.3%, respectively, P < 0.05) and increased in the SpermGrad preparation (56.3%, P < 0.05). Protamine deficiency also decreased in all three kinds of media, 59.3%, 47.7% and 40.3% for PureSperm, Sil-Select Plus and SpermGrad preparations, respectively (P < 0.05). The percentage of DNA-damaged sperm was negatively correlated with the percentages of sperm motility, rapid motility and PMC, but was positively correlated with static motility (P < 0.05). This comparative study and correlation analysis revealed that PureSperm preparation yielded sperm with the best motility and the lowest percentage of protamine deficiency. The Sil-Select Plus preparation yielded sperm with the lowest amount of DNA damage. The SpermGrad preparation had a high percentage of sperm with normal morphology, but also had the highest percentage of sperm with DNA damage. Sperm DNA damage was correlated with percentages of sperm motility, rapid motility, static motility and PMC.
Topics: Culture Media; DNA Damage; Humans; In Situ Nick-End Labeling; Male; Prospective Studies; Protamines; Semen; Sperm Motility
PubMed: 20010846
DOI: 10.1038/aja.2009.60 -
Basic & Clinical Pharmacology &... Aug 2008Anaphylactic reactions caused by injection of protamine sulfate during cardiac surgery are a well-known complication. A systematic literature review was therefore... (Review)
Review
Anaphylactic reactions caused by injection of protamine sulfate during cardiac surgery are a well-known complication. A systematic literature review was therefore conducted to gather evidence of the knowledge concerning these side effects, and to see if any prospective randomized studies supported this. Studies investigating the effect of protamine sulfate in human beings were extracted from MEDLINE, Embase and the Cochrane Library, retrieving 487 articles. Abstracts were evaluated by both authors, and referred articles not found in the primary search were furthermore extracted from reviews and case reports, resulting in a total of 272 relevant articles. Of these, 9 retrospective studies and 16 prospective studies were performed in an evidence-based manner. However, only 3 of the 16 prospective articles had an optimal design as far as inclusion criteria, randomization, and description of symptoms were concerned. Incidence of anaphylactic reactions in the prospective studies was 0.69% compared to 0.19% in the retrospective studies, but caution should be taken due to a pronounced heterogeneity of those studies. One study found heparinase I unsuitable as replacement for protamine sulfate. Overall, our findings support the low incidence of anaphylactic reactions reported in previous studies, but of note only few prospective investigations was conducted on the subject. Our study also emphasizes the need for critical appraisal of many routine procedures: in all aspects of medical care, systematic literature review conducted in a well-structured, repeated manner should be given high priority.
Topics: Anaphylaxis; Heparin Antagonists; Humans; Prospective Studies; Protamines; Randomized Controlled Trials as Topic
PubMed: 18816305
DOI: 10.1111/j.1742-7843.2008.00274.x -
Cells Jun 2020Despite the significant technical advancements in mass spectrometry-based proteomics and bioinformatics resources, dynamic resolution of soybean seed proteome is still...
In-Depth Investigation of Low-Abundance Proteins in Matured and Filling Stages Seeds of Employing a Combination of Protamine Sulfate Precipitation and TMT-Based Quantitative Proteomic Analysis.
Despite the significant technical advancements in mass spectrometry-based proteomics and bioinformatics resources, dynamic resolution of soybean seed proteome is still limited because of the high abundance of seed storage proteins (SSPs). These SSPs occupy a large proportion of the total seed protein and hinder the identification of low-abundance proteins. Here, we report a TMT-based quantitative proteome analysis of matured and filling stages seeds of high-protein (Saedanbaek) and low-protein (Daewon) soybean cultivars by application of a two-way pre-fractionation both at the levels of proteins (by PS) and peptides (by basic pH reverse phase chromatography). Interestingly, this approach led to the identification of more than 5900 proteins which is the highest number of proteins reported to date from soybean seeds. Comparative protein profiles of Saedanbaek and Daewon led to the identification of 2200 and 924 differential proteins in mature and filling stages seeds, respectively. Functional annotation of the differential proteins revealed enrichment of proteins related to major metabolism including amino acid, major carbohydrate, and lipid metabolism. In parallel, analysis of free amino acids and fatty acids in the filling stages showed higher contents of all the amino acids in the Saedanbaek while the fatty acids contents were found to be higher in the Daewon. Taken together, these results provide new insights into proteome changes during filling stages in soybean seeds. Moreover, results reported here also provide a framework for systemic and large-scale dissection of seed proteome for the seeds rich in SSPs by two-way pre-fractionation combined with TMT-based quantitative proteome analysis.
Topics: Fatty Acids; Humans; Plant Proteins; Protamines; Proteomics; Seeds; Glycine max
PubMed: 32580392
DOI: 10.3390/cells9061517 -
American Journal of Veterinary Research Apr 2014To determine the effects of protamine sulfate on clot formation time and clot strength thromboelastography variables for canine whole blood samples.
OBJECTIVE
To determine the effects of protamine sulfate on clot formation time and clot strength thromboelastography variables for canine whole blood samples.
ANIMALS
Blood samples obtained from 11 healthy dogs.
PROCEDURES
Blood samples were collected from jugular veins of dogs into syringes with 3.2% sodium citrate (blood to citrate ratio, 9:1). Blood samples were divided into aliquots, and protamine sulfate was added to various concentrations (0 [control], 22, 44, and 66 μg/mL). Prepared samples were activated with kaolin (n = 8) or not activated (8), CaCl₂ was added, and thromboelastography was performed. Reaction time (R), clot formation time (K), rate of clot formation (α angle), and maximum amplitude (MA) were measured.
RESULTS
For kaolin-activated and nonactivated blood samples, protamine (66 μg/mL) significantly increased R and K and decreased α angle and MA, compared with values for control samples. Also, protamine (44 μg/mL) decreased MA in nonactivated blood samples and increased K and decreased α angle in kaolin-activated samples, compared with values for control samples.
CONCLUSIONS AND CLINICAL RELEVANCE
Results indicated protamine prolonged clot formation time and decreased overall clot strength in a dose-dependent manner; such effects may contribute to a hypocoagulable state in dogs. Kaolin-activated and nonactivated blood samples were appropriate for measurement of the effects of protamine on coagulation. Administration of protamine to reverse the effects of heparin should be performed with caution.
Topics: Animals; Blood Coagulation; Blood Coagulation Tests; Dogs; Heparin Antagonists; Kaolin; Male; Protamines; Thrombelastography
PubMed: 24669916
DOI: 10.2460/ajvr.75.4.338 -
International Heart Journal May 2018Bleeding complications following percutaneous coronary interventions (PCI) have been closely associated with morbidity and mortality. Although radial arteries have been... (Comparative Study)
Comparative Study
Bleeding complications following percutaneous coronary interventions (PCI) have been closely associated with morbidity and mortality. Although radial arteries have been widely used in current PCI, including primary PCI, transfemoral PCI remains necessary for complex PCI. The purpose of this study was to compare the incidence of complications following elective transfemoral PCI between manual compression with and without protamine. We identified 249 consecutive patients who underwent elective transfemoral PCI from hospital records, and divided them into two groups: patients who used protamine for manual compression (the protamine group; n = 205) and patients who did not (the non-protamine group, n = 44). Complications including acute thrombosis, bleeding requiring blood transfusion, transient hypotension, skin rash, and death within 30 days were compared between groups. The baseline clinical and procedural characteristics were comparable between the protamine and non-protamine groups. The incidences of all complications were not different between the protamine (5.9%) and the non-protamine groups (9.1%) (P = 0.43). While more than 90% of the patients received drug-eluting stent implantation, there was no acute thrombus in either group. The incidence of bleeding requiring blood transfusion was significantly lower in the protamine group (0.5%) than in the non-protamine group (6.8%) (P = 0.002). Multivariate logistic regression analysis revealed the inverse association between protamine use and bleeding requiring blood transfusion (odds ratio 0.08, 95% confidence interval 0.01-0.84, P = 0.04). In conclusion, the use of protamine for manual compression following elective transfemoral PCI was safe and was associated with less bleeding complications.
Topics: Aged; Anticoagulants; Blood Transfusion; Coronary Artery Disease; Drug-Eluting Stents; Elective Surgical Procedures; Female; Hemostatic Techniques; Heparin Antagonists; Humans; Incidence; Male; Middle Aged; Percutaneous Coronary Intervention; Postoperative Complications; Protamines; Retrospective Studies; Treatment Outcome
PubMed: 29743410
DOI: 10.1536/ihj.17-352 -
The Journal of Extra-corporeal... Mar 2018Despite more than a half century of "safe" cardiopulmonary bypass (CPB), the evidence base surrounding the conduct of anticoagulation for CPB has not been organized into...
Despite more than a half century of "safe" cardiopulmonary bypass (CPB), the evidence base surrounding the conduct of anticoagulation for CPB has not been organized into a succinct guideline. For this and other reasons, there is enormous practice variability relating to the use and dosing of heparin, monitoring heparin anticoagulation, reversal of anticoagulation, and the use of alternative anticoagulants. To address this and other gaps, the Society of Thoracic Surgeons (STS), the Society of Cardiovascular Anesthesiologists (SCA), and the American Society of Extracorporeal Technology (AmSECT) developed an Evidence Based Workgroup. This was a group of interdisciplinary professionals gathered together to summarize the evidence and create practice recommendations for various aspects of CPB. To date, anticoagulation practices in CPB have not been standardized in accordance with the evidence base. This clinical practice guideline was written with the intent to fill the evidence gap and to establish best practices in anticoagulation for CPB using the available evidence. To identify relevant evidence a systematic review was outlined and literature searches were conducted in PubMed® using standardized MeSH terms from the National Library of Medicine list of search terms. Search dates were inclusive of January 2000 to December 2015. The search yielded 833 abstracts which were reviewed by two independent reviewers. Once accepted into the full manuscript review stage, two members of the writing group evaluated each of 286 full papers for inclusion eligibility into the guideline document. Ninety-six manuscripts were included in the final review. In addition, 17 manuscripts published prior to 2000 were included to provide method, context, or additional supporting evidence for the recommendations as these papers were considered sentinel publications. Members of the writing group wrote and developed recommendations based on review of the articles obtained and achieved more than two thirds agreement on each recommendation. The quality of information for a given recommendation allowed assessment of the level of evidence as recommended by the AHA/ACCF Task Force on Practice Guidelines. Recommendations were written in the three following areas 1) Heparin dosing and monitoring for initiation and maintenance of CPB, 2) Heparin contraindications and heparin alternatives, 3) Reversal of anticoagulation during cardiac operations. It is hoped that this guideline will serve as a resource and will stimulate investigators to conduct more research and expand upon the evidence base on the topic of anticoagulation for CPB.
Topics: Anticoagulants; Cardiopulmonary Bypass; Heparin; Hirudins; Humans; Peptide Fragments; Protamines; Recombinant Proteins; Societies, Medical
PubMed: 29559750
DOI: No ID Found -
International Journal of Nanomedicine 2013The objective of this research was to design an effective gene delivery system composed of cationic solid lipid nanoparticles (SLNs), protamine, and Deoxyribonucleic...
BACKGROUND
The objective of this research was to design an effective gene delivery system composed of cationic solid lipid nanoparticles (SLNs), protamine, and Deoxyribonucleic acid DNA.
METHODS
Cationic SLNs were prepared using an aqueous solvent diffusion method with octadecylamine as the cationic lipid material. First, protamine was combined with DNA to form binary protamine/DNA nanoparticles, and the ternary nanoparticle gene delivery system was then obtained by combining binary protamine/DNA nanoparticles with cationic SLNs. The size, zeta potential, and ability of the binary and ternary nanoparticles to compact and protect DNA were characterized. The effect of octadecylamine content in SLNs and the SLNS/DNA ratios on transfection efficiency, cellular uptake and cytotoxicity of the ternary nanoparticles were also assessed using HEK293 cells.
RESULTS
When the weight ratio of protamine to DNA reached 1.5:1, the plasmid DNA could be effectively compacted and protected. The average hydrodynamic diameter of the ternary nanoparticles when combined with protamine increased from 188.50 ± 0.26 nm to 259.33 ± 3.44 nm, and the zeta potential increased from 25.50 ± 3.30 mV to 33.40 ± 2.80 mV when the weight ratio of SLNs to DNA increased from 16/3 to 80/3. The ternary nanoparticles showed high gene transfection efficiency compared with Lipofectamine™ 2000/DNA nanoparticles. Several factors that might affect gene transfection efficiency, such as content and composition of SLNs, post-transfection time, and serum were examined. The ternary nanoparticles composed of SLNs with 15 wt% octadecylamine (50/3 weight ratio of SLNs to DNA) showed the best transfection efficiency (26.13% ± 5.22%) in the presence of serum. It was also found that cellular uptake of the ternary nanoparticles was better than that of the SLN/DNA and binary protamine/DNA nanoparticle systems, and DNA could be transported to the nucleus.
CONCLUSION
SLNs enhanced entry of binary protamine/DNA nanoparticles into the cell, and protamine protected DNA from enzyme degradation and transported DNA into the nucleus. Compared with Lipofectamine 2000/DNA nanoparticles, these cationic ternary nanoparticles showed relatively durable and stable gene transfection in the presence of serum.
Topics: Amines; Cations; Cell Survival; DNA; Electrophoretic Mobility Shift Assay; HEK293 Cells; Humans; Lipids; Luminescent Proteins; Microscopy, Fluorescence; Nanoparticles; Particle Size; Protamines; Transfection
PubMed: 23990715
DOI: 10.2147/IJN.S47967 -
Scientific Reports Nov 2016Protamines are arginine-rich DNA-binding proteins that replace histones in elongating spermatids. This leads to hypercondensation of chromatin and ensures physiological...
Protamines are arginine-rich DNA-binding proteins that replace histones in elongating spermatids. This leads to hypercondensation of chromatin and ensures physiological sperm morphology, thereby protecting DNA integrity. In mice and humans, two protamines, protamine-1 (Prm1) and protamine-2 (Prm2) are expressed in a species-specific ratio. In humans, alterations of this PRM1/PRM2 ratio is associated with subfertility. By applying CRISPR/Cas9 mediated gene-editing in oocytes, we established Prm2-deficient mice. Surprisingly, heterozygous males remained fertile with sperm displaying normal head morphology and motility. In Prm2-deficient sperm, however, DNA-hypercondensation and acrosome formation was severely impaired. Further, the sperm displayed severe membrane defects resulting in immotility. Thus, lack of Prm2 leads not only to impaired histone to protamine exchange and disturbed DNA-hypercondensation, but also to severe membrane defects resulting in immotility. Interestingly, previous attempts using a regular gene-targeting approach failed to establish Prm2-deficient mice. This was due to the fact that already chimeric animals generated with Prm2 ES cells were sterile. However, the Prm2-deficient mouse lines established here clearly demonstrate that mice tolerate loss of one Prm2 allele. As such they present an ideal model for further studies on protamine function and chromatin organization in murine sperm.
Topics: Animals; Base Sequence; CRISPR-Cas Systems; Exons; Female; Genetic Association Studies; Genetic Loci; Haploinsufficiency; Histones; Infertility, Male; Male; Mice; Protamines; Sequence Deletion; Sperm Motility; Spermatogenesis; Spermatozoa; Testis
PubMed: 27833122
DOI: 10.1038/srep36764 -
BioMed Research International 2022The objective of the study was to develop PEGylated protamine letrozole nanoparticles to combat human breast cancer by modifying the release pattern of letrozole. Breast...
The objective of the study was to develop PEGylated protamine letrozole nanoparticles to combat human breast cancer by modifying the release pattern of letrozole. Breast cancer is amongst the most prevalent diseases in women due to overactivity of human epidermal growth factor receptor 2 (HER2). PEG-protamine letrozole nanoparticle formulation was designed and optimized to alter the release pattern of the drug. The size, morphology, and structure of PEG-protamine letrozole NP were characterized by FTIR, XRD, Zetasizer, and SEM analysis. The result showed the PEG-protamine letrozole nanoparticles were irregular in shape and have size ranging from 258 nm to 388 nm, polydispersity index 0.114 to 0.45, zeta potential of 11.2 mV, and entrapment efficiency 89.93%. XRD studies have confirmed that the crystal structure of letrozole has become amorphous. The drug release study maintained the prolonged release for 72 hours. Moreover, the PEG-protamine letrozole NPs displayed a strong anticancer action compared to MCF-7 cells with an IC50 70 M for letrozole and 50 M for PEG-protamine letrozole NPs. Overall, our results indicate that letrozole PEG-protamine NPs alter the release profile of letrozole, which could be an excellent approach for overcoming letrozole resistance in human breast cancer.
Topics: Breast Neoplasms; Drug Carriers; Female; Humans; Letrozole; MCF-7 Cells; Nanoparticles; Particle Size; Polyethylene Glycols; Protamines
PubMed: 35722457
DOI: 10.1155/2022/4438518