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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 -
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 -
BioRxiv : the Preprint Server For... Dec 2023In sperm cells, protamine replaces histones to compact DNA 10-20 times more than in somatic cells. To characterize the extreme compaction, we employed confocal...
In sperm cells, protamine replaces histones to compact DNA 10-20 times more than in somatic cells. To characterize the extreme compaction, we employed confocal microscopy and optical tweezers to determine the conformations and stability of protamine-bound λ-DNA. Confocal images show increasing compaction of λ-DNA at increasing protamine concentration. In the presence of protamine, single λ-DNA molecules form bends and loops that unravel at 10-40 pN forces as well as coils that shorten the contour length by up to 40% and withstand forces strong enough (~55 pN) for strand separation. Strand separation nucleates coils, indicating protamine insertion into DNA bases. Protamine may participate in both local and higher-order chromatin organization, leading to extreme compaction and global transcription silencing.
PubMed: 38106194
DOI: 10.1101/2023.12.08.570784 -
Veterinary World Mar 2020Fertility is the most important aspect in the efforts to increase livestock populations. Protamine and various proteins in sperm and seminal plasma are the results of... (Review)
Review
Fertility is the most important aspect in the efforts to increase livestock populations. Protamine and various proteins in sperm and seminal plasma are the results of the molecular analysis which can be used as a marker of fertility. Each of the proteins plays an important role in the normal function of sperm, starting from the formation of sperm structure, motility, capacitation, cell protection, acrosome reactions, successful fertilization, egg activation, and embryonic development. Finally, these molecular components can be a marker of fertility and can help to diagnose the cases of infertility/subfertility in livestock in the field.
PubMed: 32367964
DOI: 10.14202/vetworld.2020.556-562 -
Andrologia Feb 2022Diabetes negatively affects the reproductive system. This present study investigated the effects of aerobic training on protamine 1 and 2 mRNA expression, sex hormones,...
Diabetes negatively affects the reproductive system. This present study investigated the effects of aerobic training on protamine 1 and 2 mRNA expression, sex hormones, antioxidant defence and sperm quality in diabetic rats. Thirty-six male Wistar rats were randomly allocated into three groups including diabetic training (DT) group, diabetic (D) group and control (C) group. Rats in DT were exercised 5 times per week for 8 weeks. Blood samples were collected for evaluation of sex hormones 48 h after the last training session. Also, the testes were removed and subjected to histological evaluation and semen analysis. Testicular mRNA expressions of protamines were determined by RT-qPCR. Protamines 1 and 2, and the ratio of protamine 1 to protamine 2 were significantly lower in DT and D groups compared with C group (p < 0.01). LH and testosterone levels were significantly lower in D group compared with DT and C group (p < 0.01). Malondialdehyde was significantly lower in DT and C groups compared with D group (p < 0.001). Sperm parameters were significantly lower in D group compared with C group (p < 0.01). Our findings suggest that aerobic training may mitigate the negative impact of diabetes on sex hormones, oxidative stress, protamine content and sperm parameters in male rats.
Topics: Animals; Diabetes Mellitus, Experimental; Fertility; Male; Rats; Rats, Wistar; Semen Analysis; Spermatozoa; Testis
PubMed: 34751459
DOI: 10.1111/and.14306 -
Andrology Nov 2022Varicocoele is a common risk factor associated with reduced male fertility potential. The current understanding of varicocoele pathophysiology does not completely... (Review)
Review
BACKGROUND
Varicocoele is a common risk factor associated with reduced male fertility potential. The current understanding of varicocoele pathophysiology does not completely explain the clinical manifestation of infertility. The present treatment options such as antioxidant supplementation and varicocoelectomy only help ≈35% of men to achieve spontaneous pregnancy.
OBJECTIVE
This review aims to summarize the available knowledge on cellular and molecular alterations implicated to varicocoele-associated male infertility and also highlights the new knowledge generated by "omics" technologies.
MATERIALS AND METHODS
PubMed, MEDLINE, Cochrane and Google Scholar databases are searched using different combinations of keywords (varicocoele, infertile/fertile men with varicocoele, cellular changes, molecular mechanisms, proteome, epigenome, transcriptome and metabolome). A total of 229 relevant human and animal studies published till 2021 were included in this review.
RESULTS
Current understanding advocates oxidative stress (OS) as a major contributory factor to varicocoele-associated male infertility. Excessive OS causes alteration in testicular microenvironment and sperm DNA fragmentation, which further contributes to infertility. Molecular and omics studies have identified several promising biomarkers such as AAMP, SPINT1, MKI67 (genetic markers), sperm quality and function related protein markers, global sperm DNA methylation level (epigenetic marker), Hspa2, Protamine, Gadd7, Dynlt1 and Beclin1 (mRNA markers), PRDX2, HSPA, APOA2, YKL40 (seminal protein markers), total choline and PHGDH (metabolic markers).
DISCUSSION
Mature spermatozoa harbours a plethora of molecular information in form of proteome, epigenome and transcriptome, which could provide very important clues regarding pathophysiology of varicocoele-associated infertility. Recent molecular and omics studies in infertile men with varicocoele have identified several promising biomarkers. Upon further validation with larger and well-defined studies, some of these biomarkers could aid in varicocoele management.
CONCLUSION
The present evidences suggest that inclusion of OS and sperm DNA fragmentation tests could be useful to the diagnostic workup for men with varicocoele. Furthermore, including precise molecular markers may assist in diagnostics and prognostics of varicocoele-associated male infertility.
Topics: Antioxidants; Beclin-1; Chitinase-3-Like Protein 1; Choline; Dyneins; Genetic Markers; Humans; Infertility, Male; Male; Protamines; Proteome; RNA, Messenger; Semen; Spermatozoa; Varicocele
PubMed: 36040837
DOI: 10.1111/andr.13278 -
Methods in Molecular Biology (Clifton,... 2021The major challenge for RNAi-based therapy is the fabrication of the delivery system that meet the requirement of clinical applicability. Liposome-derived nanoparticles...
The major challenge for RNAi-based therapy is the fabrication of the delivery system that meet the requirement of clinical applicability. Liposome-derived nanoparticles (NPs) are one of the best investigated systems for in vivo siRNA delivery. In the recent years, we have successfully redesigned the conventional cationic liposomes into Liposome/Protamine/hyaluronic acid (LPH) NPs and Lipid-Calcium-Phosphate (LCP) NPs in order to increase the in vivo gene silencing effect and reduce the toxicity. Here we describe the preparation of LPH and LCP NPs loaded with siRNA, and characterization analysis including size distribution, trapping efficiency, and in vivo activity. This protocol could be used for in vivo delivery of siRNA to target genes in cancer cells.
Topics: Animals; Calcium Phosphates; Cell Line, Tumor; Clinical Protocols; Female; Gene Transfer Techniques; Genes, Reporter; Humans; Hyaluronic Acid; Lipids; Liposomes; Luciferases; Mice, Nude; Particle Size; Protamines; RNA Interference; RNA, Small Interfering; Research Design; Mice
PubMed: 33928575
DOI: 10.1007/978-1-0716-1298-9_10 -
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