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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 -
The Journal of Extra-corporeal... Mar 2022Blood product usage is an important outcome for patients undergoing cardiac surgery. In 2015, our center made a concerted effort with multiple departments to focus on...
Blood and Blood Product Conservation: Results of Strategies to Improve Clinical Outcomes in Open Heart Surgery Patients at a Tertiary Hospital Are Maintained 4 Years after Initiation.
Blood product usage is an important outcome for patients undergoing cardiac surgery. In 2015, our center made a concerted effort with multiple departments to focus on reducing transfusion rates in surgical patients requiring cardiopulmonary bypass (CPB). Specific changes included an upgrade of the oxygenator in mid-2015 and, in early 2016, implementation of a hemostasis management system (HMS) that used heparin dose-response titration assays for heparin and protamine management. A retrospective chart review demonstrated significant decreases in the quarterly average of patients receiving packed red blood cells (PRBCs) from a baseline of 26.7% to 22.7% following the oxygenator upgrade ( = .021) and from 22.7% to 8.8% following implementation of the HMS ( = .0017). Platelet usage decreased from an average of 50.5% during the baseline and oxygenator upgrade periods to 22.2% following implementation of the HMS ( < .0001). Usage of fresh frozen plasma (FFP) decreased from an average of 28.2% of cases during the baseline and oxygenator upgrade periods to 10.4% during 2016, and cryoprecipitate usage decreased from 38.5% to 15.4%. Heparin usage averaged 56,903 units before implementation of the HMS, decreasing to an average of 43,796 units following HMS implementation ( < .0001). During the same time periods, protamine usage averaged 340.3 mg and 183.2 mg, respectively. Because improvements achieved during quality initiatives may revert back to their pre-intervention state once the assessment period is over, we performed a second retrospective analysis to determine whether the improvements achieved were maintained during the 48 months following the initial study. During 2017-2020, quarterly average usage of blood products was as follows: PRBCs, 11.9%; platelets, 14.7%; FFP, 6.2%; and cryoprecipitate, 11.5%. Quarterly, average use of heparin and protamine were 31,556 ± 2,757 units and 189 ± 113 mg, respectively. These findings indicate that the improvements achieved were not limited to the duration of the initial quality initiative.
Topics: Humans; Retrospective Studies; Tertiary Care Centers; Cardiac Surgical Procedures; Cardiopulmonary Bypass; Heparin; Protamines
PubMed: 36380821
DOI: 10.1182/ject-35-41 -
Protamine-like proteins have bactericidal activity. The first evidence in Mytilus galloprovincialis.Acta Biochimica Polonica Nov 2018The major acid-soluble protein components of the mussel Mytilus galloprovincialis sperm chromatin consist of the protamine-like proteins PL-II, PL-III and PL-IV, an...
The major acid-soluble protein components of the mussel Mytilus galloprovincialis sperm chromatin consist of the protamine-like proteins PL-II, PL-III and PL-IV, an intermediate group of sperm nuclear basic proteins between histones and protamines. The aim of this study was to investigate the bactericidal activity of these proteins since, to date, there are reports on bactericidal activity of protamines and histones, but not on protamine-like proteins. We tested the bactericidal activity of these proteins against Gram-positive bacteria: Enterococcus faecalis and two different strains of Staphylococcus aureus, as well as Gram-negative bacteria: Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella typhmurium, Enterobacter aerogenes, Enterobacter cloacae, and Escherichia coli. Clinical isolates of the same bacterial species were also used to compare their sensitivity to these proteins. The results show that Mytilus galloprovincialis protamine-like proteins exhibited bactericidal activity against all bacterial strains tested with different minimum bactericidal concentration values, ranging from 15.7 to 250 µg/mL. Furthermore, these proteins were active against some bacterial strains tested that are resistant to conventional antibiotics. These proteins showed very low toxicity as judged by red blood cell lysis and viability MTT assays and seem to act both at the membrane level and within the bacterial cell. We also tested the bactericidal activity of the product obtained from an in vitro model of gastrointestinal digestion of protamine-like proteins on a Gram-positive and a Gram-negative strain, and obtained the same results with respect to undigested protamine-like proteins on the Gram-positive bacterium. These results provide the first evidence of bactericidal activity of protamine-like-proteins.
Topics: Animals; Anti-Bacterial Agents; Gram-Negative Bacteria; Gram-Positive Bacteria; Microbial Sensitivity Tests; Mytilus; Nuclear Proteins; Protamines
PubMed: 30447153
DOI: 10.18388/abp.2018_2638 -
The Journal of Extra-corporeal... Mar 2023: Postoperative atrial fibrillation (POAF) is defined as new-onset AF in the immediate postoperative period. The relatively high incidence of POAF after cardiac surgery...
: Postoperative atrial fibrillation (POAF) is defined as new-onset AF in the immediate postoperative period. The relatively high incidence of POAF after cardiac surgery is well described, but pathophysiological mechanisms underlying the initiation, maintenance, and progression of POAF may be multifactorial and have not yet been comprehensively characterized. One of the mechanisms includes altered Ca kinetics. Accumulating evidence has suggested that altered atrial cytosolic calcium handling contributes to the development of POAF, protamine reversibly modulates the calcium release channel/ryanodine receptor 2 (RyR2) and voltage-dependent cardiac RyR2. However, it is currently unknown whether such abnormalities contribute to the arrhythmogenic substrate predisposing patients to the development of POAF. : We have retrospectively analyzed 147 patients who underwent cardiac surgery with cardiopulmonary bypass support. Of these, 40 patients were excluded from the analysis because of pre-existing AF. All patients received heparin followed by protamine at different dosing ratios of protamine-to-heparin, depending on the periods studied. : The dosing ratio of protamine-to-heparin = 1.0 was compared with higher dosing ratios of protamine-to-heparin >1.0 up to 1.7. POAF developed in 15 patients (15/107 = 14%), of these, 5 out of 57 patients (33.3%) in the dosing ratio of protamine-to-heparin = 1.0 and 10 out of 35 patients (66.7%) in the higher dosing ratios of protamine-to-heparin. Statistical significance was observed in patients with higher dosing ratios of protamine-to-heparin, compared with the dosing ratio of protamine-to-heparin = 1.0 (odds ratio = 3.890, 95% CI = 1.130-13.300, -value = 0.031). When types of diseases were analyzed in terms of higher dosing ratios of protamine-to-heparin, only valvular disorders were significantly associated with POAF ( = 0.04). : Protamine is clinically utilized to reverse heparin overdose and has been shown to display immunological and inflammatory alterations. However, its association with POAF has not been reported. Our results provide evidence that higher dosing ratios of protamine-to-heparin may increase the incidence of POAF.
Topics: Humans; Heparin; Atrial Fibrillation; Protamines; Coronary Artery Bypass; Retrospective Studies; Calcium; Ryanodine Receptor Calcium Release Channel; Postoperative Period; Postoperative Complications; Risk Factors
PubMed: 37034101
DOI: 10.1051/ject/2023003 -
Journal of Veterinary Science Jan 2019The objective of this study was to analyze the protective effects of iodixanol on dog spermatozoa during cryopreservation. The optimal concentration of iodixanol, 1.5%,...
The objective of this study was to analyze the protective effects of iodixanol on dog spermatozoa during cryopreservation. The optimal concentration of iodixanol, 1.5%, was determined using fresh spermatozoa and was applied in the following experiments. The 1.5% iodixanol group showed significantly increased sperm motility from that in the control ( < 0.05). Lower mitochondrial reactive oxygen species (ROS) modulator () and pro-apoptotic gene () expressions, together with higher expressions of protamine-2 (), protamine-3 (), anti-apoptotic B-cell lymphoma-2 (), and sperm acrosome associated-3 () genes were detected in the iodixanol-treated group. In addition, decreased protamine deficiency and cryocapacitation were observed in the treatment group. Our results show that supplementation with 1.5% iodixanol is ideal for reducing production of ROS and preventing detrimental effects during the canine sperm cryopreservation process, effects manifested as increased motility and reduced cryocapacitation in frozen-thawed spermatozoa.
Topics: Animals; Cryopreservation; DNA Damage; Dogs; Dose-Response Relationship, Drug; Male; Protamines; Protective Agents; Reactive Oxygen Species; Semen Preservation; Spermatozoa; Triiodobenzoic Acids
PubMed: 30481988
DOI: 10.4142/jvs.2019.20.1.79 -
Biophysical Journal Dec 2022Protamines are more arginine-rich and more basic than histones and are responsible for providing a highly compacted shape to the sperm heads in the testis....
Protamines are more arginine-rich and more basic than histones and are responsible for providing a highly compacted shape to the sperm heads in the testis. Phosphorylation and dephosphorylation are two events that occur in the late phase of spermatogenesis before the maturation of sperms. In this work, we have studied the effect of phosphorylation of protamine-like cationic peptides using all-atom molecular dynamics simulations. Through thermodynamic analyses, we found that phosphorylation reduces the binding efficiency of such cationic peptides on DNA duplexes. Peptide phosphorylation leads to a less efficient DNA condensation, due to a competition between DNA-peptide and peptide-peptide interactions. We hypothesize that the decrease of peptide bonds between DNA together with peptide self-assembly might allow an optimal re-organization of chromatin and an efficient condensation through subsequent peptide dephosphorylation. Based on the globular and compact conformations of phosphorylated peptides mediated by arginine-phosphoserine H-bonding, we furthermore postulate that phosphorylated protamines could more easily intrude into chromatin and participate to histone release through disruption of histone-histone and histone-DNA binding during spermatogenesis.
Topics: Male; Humans; Protamines; Histones; Phosphorylation; Semen; Chromatin; DNA; Peptides; Spermatozoa; Arginine
PubMed: 36168289
DOI: 10.1016/j.bpj.2022.09.025 -
Asian Journal of Andrology 2020Assisted reproductive technologies involving the use of spermatozoa and eggs for in vitro fertilization (IVF) have come as the solution for many infertile couples to... (Comparative Study)
Comparative Study Observational Study
Assisted reproductive technologies involving the use of spermatozoa and eggs for in vitro fertilization (IVF) have come as the solution for many infertile couples to become parents. However, in some cases, the use of ejaculated spermatozoa delivers poor IVF performance. Some studies have suggested the use of testicular spermatozoa in severe male infertility cases, but no guidelines regarding their utilization are currently available. In the present study, we found the mRNA protamine 1/protamine 2 (P1/P2) ratio to be a valuable biomarker of poor sperm function that could be used as a diagnostic key for the identification of cases that would benefit from the use of testicular spermatozoa. A total of 23 couples undergoing egg donation cycles with at least one previous cycle failure were studied. All couples underwent two consecutive intracytoplasmic sperm injection (ICSI) cycles with either ejaculated or testicular spermatozoa (TESA). The sperm mRNA P1/P2 ratio, fertilization rate, blastocyst rate, and pregnancy and live birth rate were compared. Results showed improved ICSI and clinical outcomes in cycles with testicular spermatozoa in men with altered mRNA P1/P2 ratios. TESA cycles presented significantly higher rates of fertilization (mean ± standard deviation: 76.1% ± 15.1% vs 65.5% ± 18.8%), blastocyst formation (55.0% ± 20.3% vs 30.8% ± 23.8%), and good morphological quality blastocyst (28.9% ± 22.9% vs 13.5% ± 17.9%) and also improvements on pregnancy (60.9% vs 0%) and healthy birth rates (56.5% vs 0%) than EJACULATE cycles. The results described here suggest that in patients with previous IVF/ICSI failures and aberrant mRNA protamine ratios, the use of testicular spermatozoa may be a good alternative to improve clinical outcomes.
Topics: Adult; Humans; Male; Middle Aged; Protamines; RNA, Messenger; Reproductive Techniques, Assisted; Retrospective Studies; Sperm Injections, Intracytoplasmic; Sperm Retrieval; Spermatozoa; Treatment Outcome
PubMed: 32217836
DOI: 10.4103/aja.aja_146_19 -
European Journal of Vascular and... Apr 2021
Topics: Endarterectomy, Carotid; Heparin Antagonists; Humans; Myocardial Infarction; Protamines
PubMed: 33642138
DOI: 10.1016/j.ejvs.2021.01.015 -
Journal of Colloid and Interface Science Jan 2023Combined usage of Layer-by-Layer (LbL) coating and alkaline phosphatase (ALP) - responsive charge reversal strategies can improve the cellular internalisation of the...
HYPOTHESIS
Combined usage of Layer-by-Layer (LbL) coating and alkaline phosphatase (ALP) - responsive charge reversal strategies can improve the cellular internalisation of the colloidal drug delivery systems by also decreasing their cytotoxic effects.
EXPERIMENTS
Anionic core NLCs were formed by combining the melt emulsification method and ultrasonication. The resulting core NLCs were coated sequentially first with protamine (Prot NLCs) and then with sodium tripolyphosphate (TPP) or sodium polyphosphate (Graham's salt, PP) generating TPP or PP NLCs, respectively. The developed NLCs were characterised regarding their size and zeta potential. Enzyme-induced charge reversal of the TPP and PP NLCs was evaluated by zeta potential measurements upon their incubation with alkaline phosphatase (ALP). In parallel, time-dependent phosphate release was monitored in the presence of isolated as well as cell-associated ALP. Morphological evaluations were performed by scanning electron microscopy (SEM) studies. Moreover, cell viability and cellular uptake studies were carried out in vitro on Caco-2 cells.
FINDINGS
The core NLCs were obtained with a mean size of 272.27 ± 5.23 nm and a zeta potential of -25.70 ± 0.26 mV. Upon coating with protamine, the zeta potential raised to positive values with a total change up to Δ29.3 mV also displaying an increase in particle size. The second layer coating with TPP and PP provided a negative surface charge. Subsequent to ALP treatment, the zeta potential of the TPP and PP NLCs reversed from negative to positive values with total changes of Δ8.56 and Δ7.47 mV, respectively. Conformably, significant amounts of phosphate were released from both formulations. Compared with core NLCs, improved cellular viability as well as increased cellular uptake were observed in case of Prot, TPP and PP NLCs.
Topics: Humans; Drug Carriers; Caco-2 Cells; Lipids; Alkaline Phosphatase; Nanostructures; Particle Size; Polyphosphates; Protamines; Sodium
PubMed: 36088699
DOI: 10.1016/j.jcis.2022.08.190 -
The Journal of Thoracic and... Oct 2015Platelet-activating antibodies against protamine-heparin-complexes were described in patients undergoing cardiac surgery, but their clinical consequences remain unclear.... (Observational Study)
Observational Study
OBJECTIVES
Platelet-activating antibodies against protamine-heparin-complexes were described in patients undergoing cardiac surgery, but their clinical consequences remain unclear. This prospective single-center observational study aimed to describe the prevalence and clinical consequences of protamine-heparin-complex antibodies in patients undergoing cardiac surgery with cardiopulmonary bypass.
METHODS
A total of 200 patients undergoing cardiac surgery with cardiopulmonary bypass were included. Blood samples were collected preoperatively and 1 hour, 24 hours, and 7 days after weaning from cardiopulmonary bypass. All sera were tested for the presence of protamine-heparin-complex antibodies using a modified heparin-induced platelet-activation assay. Specific Fcγ receptor IIa-dependent platelet activation was confirmed by repeated testing in the presence of the Fcγ receptor IIa-blocking antibody IV.3.
RESULTS
Samples from 185 patients were obtained, of whom 24 patients (13%) were positive for protamine-heparin-complex antibodies preoperatively. In all positive samples, functional reactivity was reversible in the presence of IV.3. Although patients with a preoperative presence of protamine-heparin-complex antibodies were significantly older compared with patients negative for protamine-heparin-complex antibodies (73 ± 9.8 years vs 68 ± 10 years, P = .037), no other potential risk factors were identified at 1 day before operation. Patients with protamine-heparin-complex antibodies required significantly more protamine to neutralize heparin (47.66 mg vs 41.67 mg, P = .027). Protamine-heparin-complex antibodies have no significant influence on perioperative platelet numbers, bleeding complications, transfusion requirement, thromboembolic events, major cardiovascular and cerebrovascular events, inflammation parameters, or kidney function.
CONCLUSIONS
Protamine-heparin-complex antibodies occur frequently in patients undergoing cardiac surgery on cardiopulmonary bypass, resulting in specific platelet activation in vitro. Protamine-heparin-complex antibodies are associated with increased protamine requirement after cardiopulmonary bypass and possibly slower recovery of platelet numbers.
Topics: Aged; Antibodies; Anticoagulants; Cardiopulmonary Bypass; Female; Heparin; Heparin Antagonists; Humans; Male; Platelet Activation; Prospective Studies; Protamines
PubMed: 26298870
DOI: 10.1016/j.jtcvs.2015.07.057