Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separa...Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.展开更多
Protamine was investigated for its antibacterial activity against the periodontal pathogens, Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans. We determined the minimum inhibit...Protamine was investigated for its antibacterial activity against the periodontal pathogens, Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans. We determined the minimum inhibitory concentrations of protamine and its hydrolysate and their bactericidal activity. Protamine inhibited the growth of all periodontopathic bacteria tested on agar plates. Protamine, which MIC was 6.3 × 10-7 g L-1, was most effective against P. gingivalis. The antibacterial effect of native protamine was higher than that of its hydrolysate. An ATP bioluminescence assay revealed that protamine showed bactericidal activity against P. gingivalis in a time-dependent manner. These results indicate that protamine could be candidate peptide for prevention of P. gingivalis infection.展开更多
Objective:To study the influence of N-acetylcysteine on the pituitary-gonadal axis hormones and protamine expression level in streptozotocin-induced diabetic male rats.Methods:Forty-two adult male Wistar rats were div...Objective:To study the influence of N-acetylcysteine on the pituitary-gonadal axis hormones and protamine expression level in streptozotocin-induced diabetic male rats.Methods:Forty-two adult male Wistar rats were divided into 6 groups,with 7 rats in each group.The control group left untreated;the streptozotocin group only received 50 mg/kg body weight streptozotocin intraperitoneally for 5 days to induce diabetes;the N-acetylcysteine group only received 200 mg/kg body weight N-acetylcysteine intraperitoneally,and the streptozotocin+N-acetylcysteine groups 1,2 and 3 received 50 mg/kg streptozotocin intraperitoneally for 5 days to induce diabetes and then received 100,200 and 400 mg/kg body weight doses of N-acetylcysteine intraperitoneally for 28 days,respectively.Enzyme-linked immunosorbent assay was used to measure the serum levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH)and testosterone,and real-time PCR was applied for measuring protamine expression level.Results:Compared to the control and N-acetylcysteine groups,a significant decrease in the body weight,testicular weight and levels of testosterone and protamine expression was observed in the streptozotocin group and the streptozotocin+N-acetylcysteine groups 1 and 2.On the contrary,the levels of LH and FSH increased significantly.In the streptozotocin+N-acetylcysteine group 3,the body weight,testicular weight and expression level of protamine were significantly higher than those of the streptozotocin group.In the streptozotocin+N-acetylcysteine groups,testosterone and LH levels were significantly higher than and lower than the streptozotocin group,respectively.In the streptozotocin+N-acetylcysteine groups 2 and 3,the level of FSH was significantly lower than that of the streptozotocin group and streptozotocin+N-acetylcysteine group 1.Furthermore,a significant increase in the expression level of protamine was observed in the streptozotocin+N-acetylcysteine groups 2 and 3 when compared to the streptozotocin group and streptozotocin+N-acetylcysteine group 1.Conclusions:N-acetylcysteine in an optimal dose of 400 mg/kg body weight has a protective influence on the pituitary-gonadal axis hormones and also on the expression level of protamine in diabetic male rats.展开更多
In recent years, various physiological functions of salmon milt extract, which consists of nucleic acid and nucleoprotein, have been reported. The objective of this study is to analyze the physiological function and i...In recent years, various physiological functions of salmon milt extract, which consists of nucleic acid and nucleoprotein, have been reported. The objective of this study is to analyze the physiological function and its mechanism of salmon milt extract (NG) on nematodes (C. elegans). The wild type nematode N2 strain was bred on the plate containing of NG for four days, and its body length increased depending on NG concentration. When nematodes were bred with NG for a longer period, average lifespan was increased, and survival rate was increased by up to 20%. Generally, the movement of nematodes decreases with longer breeding period (i.e. aging). Analysis of movement (both gross thrashing movement and local pumping movement) showed that NG suppressed this decrease f movement with aging. Furthermore, the deease of survival rate by heat stress and oxidative stress was suppressed by NG administration. Nile Red staining analysis showed that fat accumulation varied depending on the concentration of NG. RT-PCR analysis revealed that the mRNA expression levels of the stress resistance genes sod-3 and sod-4 were increased. These results indicated that NG administration increased the expression of stress-tolerance-related genes, promoted stress tolerance, increased movement and prolonged lifespan in nematode.展开更多
An analysis of selected datasets from the FDA’s drug Adverse Event Reporting System (FAERS) leads us to hypothesize that glyphosate contamination in both food and drugs is a major contributor to chronic and acute kid...An analysis of selected datasets from the FDA’s drug Adverse Event Reporting System (FAERS) leads us to hypothesize that glyphosate contamination in both food and drugs is a major contributor to chronic and acute kidney failure respectively. In chronic kidney failure, glyphosate-induced pancreatitis results in the release of trypsin, causing a leaky vasculature. The albumin-bound glyphosate escapes into the tissues, protecting the circulatory system and kidneys but resulting in multiple symptoms related to skin, gut, brain, bones, lungs, etc. The rare and poorly understood acute kidney failure response reported for protamine sulfate and Trasylol? is strikingly similar to that associated with glyphosate poisoning. Both drugs are derived from biological tissues that are plausibly contaminated with glyphosate. These drugs protect from haemorrhage, which leads to retention of glyphosate in the vasculature, are followed by circulatory collapse and a high likelihood of death as an outcome. We support our argument by comparing symptom profiles of selected subsets of FAERS with those related to glyphosate poisoning, anomalous reactions to protamine sulfate, and conditions showing strong statistical time-trend correlations with glyphosate.展开更多
<div style="text-align:justify;"> <span style="font-family:Verdana;">Type 2 diabetes mellitus is a growing health problem, characterized by insulin resistance progressing to beta cell d...<div style="text-align:justify;"> <span style="font-family:Verdana;">Type 2 diabetes mellitus is a growing health problem, characterized by insulin resistance progressing to beta cell dysfunction and insulin deficiency, most of these patients will need intensification of treatment and initiation of insulin to delay or prevent diabetic complications. Glycemic control is the most important aspect of management, and in reducing morbidity and mortality of the diseases. Control of plasma glucose in patients with diabetes can be assessed by HbA1c, FPG, PPG, but still HbA1c% remains the gold standard for assessment of glycemic control and follow up of diabetic patients. The aim of this study is to assess HbA1c% in patients on oral anti-diabetic drugs, with poor glycemic control before and after adding basal insulin, with titration of the dose of insulin depending on fasting blood sugar. 82 patients with uncontrolled type 2 diabetes (43.9% male, 56.1% female), with HbA1c more than 9%, on two types of oral diabetic medication or more, were started on basal insulin (glargine, lantus) and followed for three to six months. Overall 82 patients with type 2 diabetes mellitus were included in the study. The mean age of the study population was 58.4 years, the mean duration of the disease range was 13.4 years. All patients with HbA1c more than 9%, without organ failure, were included in the study. The mean HbA1c overall had decreased from mean of 11.15% before starting basal insulin to the mean of 8.43% within 3 to 6 month, after initiating basal insulin, this difference was significant at p < 0.001. There was no adverse effect on this medication in any of the study group. The addition of basal insulin to oral anti-diabetic medication in uncontrolled insulin-na<span style="color:#4F4F4F;font-family:"font-size:14px;white-space:normal;background-color:#F7F7F7;">ï</span>ve type 2 diabetic patients resulted in significant improvement of glycemic control, with improved HbA1c level, without adverse effects.</span> </div>展开更多
Brain delivery of macromolecular therapeutics(e.g., proteins) remains an unsolved problem because of the formidable blood–brain barrier(BBB). Although a direct pathway of nose-to-brain transfer provides an answer to ...Brain delivery of macromolecular therapeutics(e.g., proteins) remains an unsolved problem because of the formidable blood–brain barrier(BBB). Although a direct pathway of nose-to-brain transfer provides an answer to circumventing the BBB and has already been intensively investigated for brain delivery of small drugs,new challenges arise for intranasal delivery of proteins because of their larger size and hydrophilicity. In order to overcome the barriers and take advantage of available pathways(e.g., epithelial tight junctions, uptake by olfactory neurons, transport into brain tissues, and intra-brain diffusion), a low molecular weight protamine(LMWP) cell-penetrating peptide was utilized to facilitate nose-to-brain transport. Cell-penetrating peptides(CPP)have been widely used to mediate macromolecular delivery through many kinds of biobarriers. Our results show that conjugates of LMWP–proteins are able to effectively penetrate into the brain after intranasal administration.The CPP-based intranasal method highlights a promising solution for protein therapy of brain diseases.展开更多
Assisted reproductive technologies invoIving 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 ejacu...Assisted reproductive technologies invoIving 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 in fertility cases,but no guideli nes 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.展开更多
Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation.Yet,its effects on sperm cytoplasmic maturation remain largely unexplored.The relationships of protamines,sperm morpholo...Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation.Yet,its effects on sperm cytoplasmic maturation remain largely unexplored.The relationships of protamines,sperm morphology,DNA damage,and cytoplasmic remodeling were illustrated in this study to provide fresh perspectives on the mechanisms of male infertility.A total of 205 infertile males were allocated into 5 groups according to the percentage of spermatozoa exhibiting abnormal morphology within their samples.Sperm concentration,motility,abnormal sperm morphology,cytoplasmic droplets(CDs),and excess residual cytoplasm(ERC)were analyzed according to the World Health Organization manual(2010).Sperm nuclear vacuoles(NVs)were determined by propidium iodide(PI)staining.Sperm protamine expressions(P1 and P2)were detected by western blot.DNA damage was measured by acridine orange test(AOT)to calculate the proportion of sperm with single-strand DNA breaks(SSBs).Our data showed that sperm concentration and motility in infertile males significantly decreased with the severity of abnormal sperm morphology(both P<0.01).P1 level,P1/P2 ratio,and SSB rate increased with the severity of sperm dysmorphology,whilst the P2 level decreased(all P<O.01).NVs,CDs,and ERC were more common in males with sperm dysmorphology and positively correlated with the SSB rate(all P<O.01).The relationships between the SSB rate and the P1/P2 ratio were also significant(P<0.01).Aberrant protamination may cause sperm dysmorphology and compromise male fertility by impairing sperm's nucleus and cytoplasm maturation,with the P1/P2 ratio potentially serving as a valuable indicator of sperm quality and male fertility.展开更多
基金This research was supported by the European Union’s Horizon 2020 research and innovation scheme under the Marie Skłodowska-Curie grant agreement No.801342(Tecniospring INDUSTRYGrant:TECSPR-19-1-0003)+4 种基金the Ministry of Science and Innovation,Spain(Grants:PID2020-113320RB-I00,PID2020-113493RB-I00,RYC2021-034546-I and RYC2021-034764-I)the Catalan Agency for Management of University and Research Grants,Regional Government of Catalonia,Spain(Grants:2017-SGR-1229 and 2021-SGR-00900)the Seneca Foundation,Regional Government of Murcia,Spain(Grant:21935/PI/22)La Marato de TV3 Foundation(Grant:214/857-202039)and the Catalan Institution for Research and Advanced Studies(ICREA).
文摘Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.
文摘Protamine was investigated for its antibacterial activity against the periodontal pathogens, Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans. We determined the minimum inhibitory concentrations of protamine and its hydrolysate and their bactericidal activity. Protamine inhibited the growth of all periodontopathic bacteria tested on agar plates. Protamine, which MIC was 6.3 × 10-7 g L-1, was most effective against P. gingivalis. The antibacterial effect of native protamine was higher than that of its hydrolysate. An ATP bioluminescence assay revealed that protamine showed bactericidal activity against P. gingivalis in a time-dependent manner. These results indicate that protamine could be candidate peptide for prevention of P. gingivalis infection.
文摘Objective:To study the influence of N-acetylcysteine on the pituitary-gonadal axis hormones and protamine expression level in streptozotocin-induced diabetic male rats.Methods:Forty-two adult male Wistar rats were divided into 6 groups,with 7 rats in each group.The control group left untreated;the streptozotocin group only received 50 mg/kg body weight streptozotocin intraperitoneally for 5 days to induce diabetes;the N-acetylcysteine group only received 200 mg/kg body weight N-acetylcysteine intraperitoneally,and the streptozotocin+N-acetylcysteine groups 1,2 and 3 received 50 mg/kg streptozotocin intraperitoneally for 5 days to induce diabetes and then received 100,200 and 400 mg/kg body weight doses of N-acetylcysteine intraperitoneally for 28 days,respectively.Enzyme-linked immunosorbent assay was used to measure the serum levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH)and testosterone,and real-time PCR was applied for measuring protamine expression level.Results:Compared to the control and N-acetylcysteine groups,a significant decrease in the body weight,testicular weight and levels of testosterone and protamine expression was observed in the streptozotocin group and the streptozotocin+N-acetylcysteine groups 1 and 2.On the contrary,the levels of LH and FSH increased significantly.In the streptozotocin+N-acetylcysteine group 3,the body weight,testicular weight and expression level of protamine were significantly higher than those of the streptozotocin group.In the streptozotocin+N-acetylcysteine groups,testosterone and LH levels were significantly higher than and lower than the streptozotocin group,respectively.In the streptozotocin+N-acetylcysteine groups 2 and 3,the level of FSH was significantly lower than that of the streptozotocin group and streptozotocin+N-acetylcysteine group 1.Furthermore,a significant increase in the expression level of protamine was observed in the streptozotocin+N-acetylcysteine groups 2 and 3 when compared to the streptozotocin group and streptozotocin+N-acetylcysteine group 1.Conclusions:N-acetylcysteine in an optimal dose of 400 mg/kg body weight has a protective influence on the pituitary-gonadal axis hormones and also on the expression level of protamine in diabetic male rats.
文摘In recent years, various physiological functions of salmon milt extract, which consists of nucleic acid and nucleoprotein, have been reported. The objective of this study is to analyze the physiological function and its mechanism of salmon milt extract (NG) on nematodes (C. elegans). The wild type nematode N2 strain was bred on the plate containing of NG for four days, and its body length increased depending on NG concentration. When nematodes were bred with NG for a longer period, average lifespan was increased, and survival rate was increased by up to 20%. Generally, the movement of nematodes decreases with longer breeding period (i.e. aging). Analysis of movement (both gross thrashing movement and local pumping movement) showed that NG suppressed this decrease f movement with aging. Furthermore, the deease of survival rate by heat stress and oxidative stress was suppressed by NG administration. Nile Red staining analysis showed that fat accumulation varied depending on the concentration of NG. RT-PCR analysis revealed that the mRNA expression levels of the stress resistance genes sod-3 and sod-4 were increased. These results indicated that NG administration increased the expression of stress-tolerance-related genes, promoted stress tolerance, increased movement and prolonged lifespan in nematode.
文摘An analysis of selected datasets from the FDA’s drug Adverse Event Reporting System (FAERS) leads us to hypothesize that glyphosate contamination in both food and drugs is a major contributor to chronic and acute kidney failure respectively. In chronic kidney failure, glyphosate-induced pancreatitis results in the release of trypsin, causing a leaky vasculature. The albumin-bound glyphosate escapes into the tissues, protecting the circulatory system and kidneys but resulting in multiple symptoms related to skin, gut, brain, bones, lungs, etc. The rare and poorly understood acute kidney failure response reported for protamine sulfate and Trasylol? is strikingly similar to that associated with glyphosate poisoning. Both drugs are derived from biological tissues that are plausibly contaminated with glyphosate. These drugs protect from haemorrhage, which leads to retention of glyphosate in the vasculature, are followed by circulatory collapse and a high likelihood of death as an outcome. We support our argument by comparing symptom profiles of selected subsets of FAERS with those related to glyphosate poisoning, anomalous reactions to protamine sulfate, and conditions showing strong statistical time-trend correlations with glyphosate.
文摘<div style="text-align:justify;"> <span style="font-family:Verdana;">Type 2 diabetes mellitus is a growing health problem, characterized by insulin resistance progressing to beta cell dysfunction and insulin deficiency, most of these patients will need intensification of treatment and initiation of insulin to delay or prevent diabetic complications. Glycemic control is the most important aspect of management, and in reducing morbidity and mortality of the diseases. Control of plasma glucose in patients with diabetes can be assessed by HbA1c, FPG, PPG, but still HbA1c% remains the gold standard for assessment of glycemic control and follow up of diabetic patients. The aim of this study is to assess HbA1c% in patients on oral anti-diabetic drugs, with poor glycemic control before and after adding basal insulin, with titration of the dose of insulin depending on fasting blood sugar. 82 patients with uncontrolled type 2 diabetes (43.9% male, 56.1% female), with HbA1c more than 9%, on two types of oral diabetic medication or more, were started on basal insulin (glargine, lantus) and followed for three to six months. Overall 82 patients with type 2 diabetes mellitus were included in the study. The mean age of the study population was 58.4 years, the mean duration of the disease range was 13.4 years. All patients with HbA1c more than 9%, without organ failure, were included in the study. The mean HbA1c overall had decreased from mean of 11.15% before starting basal insulin to the mean of 8.43% within 3 to 6 month, after initiating basal insulin, this difference was significant at p < 0.001. There was no adverse effect on this medication in any of the study group. The addition of basal insulin to oral anti-diabetic medication in uncontrolled insulin-na<span style="color:#4F4F4F;font-family:"font-size:14px;white-space:normal;background-color:#F7F7F7;">ï</span>ve type 2 diabetic patients resulted in significant improvement of glycemic control, with improved HbA1c level, without adverse effects.</span> </div>
基金the National Basic Research Program of China(973 Program Nos.2013CB932503 and 2014CB931900)National Natural Science Foundation of China(Nos.81172996,81373357,81422048 and 81361140344)
文摘Brain delivery of macromolecular therapeutics(e.g., proteins) remains an unsolved problem because of the formidable blood–brain barrier(BBB). Although a direct pathway of nose-to-brain transfer provides an answer to circumventing the BBB and has already been intensively investigated for brain delivery of small drugs,new challenges arise for intranasal delivery of proteins because of their larger size and hydrophilicity. In order to overcome the barriers and take advantage of available pathways(e.g., epithelial tight junctions, uptake by olfactory neurons, transport into brain tissues, and intra-brain diffusion), a low molecular weight protamine(LMWP) cell-penetrating peptide was utilized to facilitate nose-to-brain transport. Cell-penetrating peptides(CPP)have been widely used to mediate macromolecular delivery through many kinds of biobarriers. Our results show that conjugates of LMWP–proteins are able to effectively penetrate into the brain after intranasal administration.The CPP-based intranasal method highlights a promising solution for protein therapy of brain diseases.
基金The authors thank all the patients for consenting to participate in this study.Furthermore,the technical assistance of Barbara Frohlich and Mareike Buch-Heberling is gratefully acknowledged.KS was supported by a Research Grant from the University Medical Center Giessen and Marburg(UKGM,project 29/2015GI).This research did not receive any other specific grant from funding agencies in the public,commercial,or not-for-profit sectors.
文摘Assisted reproductive technologies invoIving 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 in fertility cases,but no guideli nes 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.
文摘Aberrant sperm protamination is linked to sperm dysmorphology and nuclear chromatin condensation.Yet,its effects on sperm cytoplasmic maturation remain largely unexplored.The relationships of protamines,sperm morphology,DNA damage,and cytoplasmic remodeling were illustrated in this study to provide fresh perspectives on the mechanisms of male infertility.A total of 205 infertile males were allocated into 5 groups according to the percentage of spermatozoa exhibiting abnormal morphology within their samples.Sperm concentration,motility,abnormal sperm morphology,cytoplasmic droplets(CDs),and excess residual cytoplasm(ERC)were analyzed according to the World Health Organization manual(2010).Sperm nuclear vacuoles(NVs)were determined by propidium iodide(PI)staining.Sperm protamine expressions(P1 and P2)were detected by western blot.DNA damage was measured by acridine orange test(AOT)to calculate the proportion of sperm with single-strand DNA breaks(SSBs).Our data showed that sperm concentration and motility in infertile males significantly decreased with the severity of abnormal sperm morphology(both P<0.01).P1 level,P1/P2 ratio,and SSB rate increased with the severity of sperm dysmorphology,whilst the P2 level decreased(all P<O.01).NVs,CDs,and ERC were more common in males with sperm dysmorphology and positively correlated with the SSB rate(all P<O.01).The relationships between the SSB rate and the P1/P2 ratio were also significant(P<0.01).Aberrant protamination may cause sperm dysmorphology and compromise male fertility by impairing sperm's nucleus and cytoplasm maturation,with the P1/P2 ratio potentially serving as a valuable indicator of sperm quality and male fertility.