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Copyright of Revista Cientifica de la Facultade de Veterinaria is the property of Universidad del Zulia, Facultad de Ciencias Veterinarias and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In this study annual fluctuations of DNA fragmentation and quality of cold-stored equine sperm were evaluated. Ejaculates were collected weekly during one year from 15 stallions. Ejaculate volume, sperm concentration and total sperm count were determined and semen was then extended and cold-stored for 48 h. Sperm motility was evaluated by CASA before and after 24 as well as 48 h of cold storage. In addition, the percentages of sperm with intact plasma membrane and acrosome (PMAI %) and with low intracellular Ca 2+ level were determined in cold-stored semen (24 h, 48 h). SCSA™ was performed to assess mean DFI, SD of DFI and % DFI in raw frozen-thawed as well as in extended sperm after 24 and 48 h of storage. The month of semen collection affected (P < 0.05) all parameters evaluated in raw semen and all criteria except progressive motility as well as rapid cells in semen stored for 24 and 48 h, respectively. Ejaculate volume was higher and sperm concentration lower in summer compared to winter and motility lower in July than in any other month of the year (P < 0.05). In semen processed in April and stored for 24 h the percentage of rapid cells was improved compared to January and after 48 h of storage progressive motility (%) was higher in January and October than in July (P < 0.05). After 24 h of cold storage PMAI % was higher in October than in January and after 48 h values were higher in September compared to January and February as well as from April to July (P < 0.05). Regarding sperm with low intracellular Ca +2 level (%) after storage for 24 and 48 h, higher values were measured in winter and in October compared to April, June and July (P < 0.01). Seasonal changes in DNA fragmentation were most evident with respect to mean DFI. In raw frozen-thawed semen mean DFI was lower from August to November than in June and July (P < 0.001). Values were lower during winter compared to spring and early summer (P < 0.05) and lower in December than from April to September (P < 0.001). After 24 h of cold storage mean DFI was lower in September and October when compared to January, February, May, July and November (P < 0.05) and after 48 h storage mean DFI was reduced in spring and autumn compared to February, June and July (P < 0.05). In conclusion, a seasonal effect was evident on semen characteristics of raw and cold-stored sperm. Semen quality was impaired in midsummer when low sperm motility and viability were combined with an elevated DNA fragmentation and Ca 2+ level of sperm. [ABSTRACT FROM AUTHOR]

Background: Teratoasthenozoospermia (TA) is a severe form of male infertility with no clear etiology. Objective: To compare the level of intracellular anion superoxide (O2-), heat shock protein A2 (HSPA2) and protamine deficiencies in ejaculated spermatozoa between teratoasthenozoospermic and normozoospermic men. Materials and Methods: In this case- control study, semen samples of 20 infertile men, with TA (with normal morphology lower than 4%_ and total motility lower than 40% ) as the case group and 20 normozoospermic fertile men as the control group were evaluated for intracellular O2- and HSPA2 by flow cytometry and protamine deficiency by Chromomycin A3 (CMA3) test. Results: The rate of CMA3+ spermatozoa in the case group was higher than controls (p=0.001). The percentages of HSPA2+ spermatozoa in the cases were significantly lower than controls (p=0.001). Also, intracellular O2- levels in the case group were significantly higher than controls (p=0.001) and had positive correlations with sperm apoptosis (r=0.79, p=0.01) and CMA3 positive sperm (r=0.76, p=0.01), but negative correlations with normal morphology (r=-0.81, p=0.01) and motility (r=-0.81, p=0.01). There was no significant correlation between intracellular O2- and HSPA2 in the case group (r=0.041, p=0.79). Conclusion: We suggest that the increase in intracellular O2-, decrease in spermatozoa HSPA2+, and high percentages of spermatozoa with immature chromatin might be considered as etiologies of infertility in TA patients. [ABSTRACT FROM AUTHOR]

In this review we provide an overview of the components of the spermatozoa playing an important role in reproductive success beyond fertilization, showing the relationship between the integrity of the diverse elements and the development of a healthy offspring. The present knowledge about fish sperm chromatin organization, epigenetic modifications of DNA and histones and sperm-borne RNAs, essential in controlling embryo development, is summarized, pointing out the possibility of using specific genes or transcripts as biomarkers of sperm quality. Data about commercial species are reported when available and more detailed information about zebrafish sperm is presented. Considering the implications that the integrity of sperm genome and epigenome has on the preservation of a proper genotype and phenotype in the progeny, the methods applied for the study of chromatin damage and for the study of transcriptome are described. Moreover we discuss some injuring agents affecting paternal information, from the presence of contaminants in the aquatic environment, to the reproductive practices applied in fish farming. The consequences of fertilizing with damaged spermatozoa, as well as the zygotic ability to repair damage are also reviewed. Statement of relevance Role of the sperm on achieving high rates of healthy fries. [ABSTRACT FROM AUTHOR]

Background: Methamphetamine (MA) was shown to have harmful effects on male reproductive system. Objective: To investigate probable effects of daily administration of MA on sperm parameters and chromatin/DNA integrity in mouse. Material and Methods: Thirty-five NMRI male mice were divided into five groups including low, medium, and high dosage groups which were injected intraperitoneally with 4, 8 and 15 mg/kg/day for 35 days, respectively. Normal saline was injected in sham group and no medications were used in control group. Then, the mice were killed and caudal epididymis of each animal was cut and placed in Ham's F10 medium for sperm retrieval. To evaluate sperm chromatin abnormalities, the aniline blue, toluidine blue and chromomycine A3 were used. For sperm DNA integrity and apoptosis, the acridine orange, sperm chromatin dispersion, and TUNEL assay were applied. For sperm morphology, Papanicolaou staining was done. Results: Normal morphology and progressive motility of spermatozoa decreased in medium and high dosage groups in comparison with the control group (p=0.035). There was a significant increase in rate of aniline blue, toluidine blue, and chromomycine A3 positive spermatozoa in high dosage group. In a similar manner, there was an increase in rates of acridine orange, TUNEL and sperm chromatin dispersion positive sperm cells in high dosage group with respect to others. Conclusion: MA abuse in a dose-dependent manner could have detrimental effects on male reproductive indices including sperm parameters and sperm chromatin/DNA integrity in mice. [ABSTRACT FROM AUTHOR]

The study of the sperm proteins is crucial for understanding its normal function and alterations in infertile patients. The sperm is a highly specialized cell with a very large flagella, with little cytoplasm and a highly condensed nucleus. The most abundant proteins in the nucleus of mammalian sperm are the protamines. The main functions of the protamines are the condensation of the DNA, possibly contributing to the generation of a more hydrodynamic sperm head and to the protection of the genetic message. However, in addition to protamines, about 5.0-15.0% of the paternal genome is also complexed with histones and histone variants. It has also demonstrated a differential distribution of genes in regions associated with histone and protamine-associated regions, suggesting a potential epigenetic relevance in embryonic development. More recently, detailed lists of proteins have been described corresponding to the different compartments of the sperm cell thanks to the application of recent proteomic techniques based on mass spectrometry (MS). Differential proteomics is also being applied to identify the presence of protein abnormalities found in infertile patients

The aim of this study was to investigate the effect of boar age on quality traits and fertility of liquid-stored semen. Boars were allocated into 3 age groups: 7-10 months (young), 18-33 months (mature), 51-61 months (old). Ejaculates of > 200x10(6) sperm/ml and 85% total motile sperm were extended to 30x10(6) sperm/ml, stored at 17-18 °C and used within 12-24 h for artificial insemination (AI) of 2062 multiparous sows. After 24 h of storage, aliquots of diluted semen were assessed for sperm progressive motility (SPM), incidence of sperm chromatin instability (SCI), proportion of live morphologically normal sperm (LMNS) and head morphometry of LMNS. The results showed that young boars had higher percentages of SCI and lower proportions of LMNS than those of the mature (p < 0.05) and old (p < 0.001) boars, respectively. Sperm head dimensions of young and old boars were greater (p < 0.03-0.001) than those of mature boars. The farrowing rate of young boars (65%) was significantly lower (p < 0.001; χ2= 30-61) than those of the mature (87.2%) and old (84.7%) boars. The relationship between sperm head dimensions and boar fertility was non-significant. In conclusion, boar age is an important physiological factor contributing to the success of swine AI.

The aim of this study was to determine the percentage of sperm with damaged chromatic measure with toluidine blue stain and it´s relationship with motility and viability in criopreserverd semen from Brahman bulls. Three ejaculates from six Brahman bulls were used. Immediately after thawing, sperms were stained with toluidine blue to establish chromatin integrity (sperms with normal chromatin were light blue or green while sperms with damaged chromatin were dark blue or violet). Sperms were also stained with eosin-nigrosin to determine viability (live sperms were unstained while dead sperms were pink). Motility was measured under light microscope. Effects of bull, ejaculate, and the interaction between variables were assessed. The percentage of live sperms was 50.02 ( ± 14.13%). The mean motility was 33.88 (± 12.43%), while the percentage of sperms with damaged chromatin was 4.17 ( ± 2.96%). Viability was positively correlated with motility (r=0.77217, p=0.0002), and negatively correlated with damaged chromatin sperms (r= -0.43104, p=0.0087). Motility percentage was negatively correlated with the percentage of sperms with damaged chromatin (r=-0.48337, p=0.0421). In conclusion, cryopreserved semen of Brahman bulls presented a low level of chromatin damage, and this trait was negatively correlated with sperm motility and viability.El objetivo del presente trabajo fue determinar el porcentaje de espermatozoides con cromatina dañada medida con la tinción de azul de toluidina, y su relación con la motilidad y la vitalidad del semen criopreservado de toros Brahma. Para ello, se utilizó semen de tres eyaculados de seis toros Brahman, el cual una vez descongelado se procedió a teñir con azul de toluidina para determinar la integridad de la cromatina (espermatozoides con cromatina normal teñidos de azul o verde claro; espermatozoides con cromatina anormal teñidos de azul oscuro o violeta), también se tiñeron con eosinanigrosina para determinar la viabilidad (espermatozoides vivos permanecen blancos; espermatozoides muertos se tiñen de rosado) y se estimó la motilidad espermática mediante microscopía óptica. Se evidenciaron las diferencias en todos los parámetros evaluados debidas al efecto toro y al eyaculado, así como a la interacción entre estas dos variables. El porcentaje de espermatozoides vivos fue de 50.02 ± 14.13% y la motilidad espermática promedió un 33.88 ± 12.43%, mientras que el porcentaje de espermatozoides con cromatina dañada fue de 4.17 ± 2.96%. El porcentaje de espermatozoides vivos se correlacionó positivamente con la motilidad (r=0.77217, p=0.0002), y negativamente con el porcentaje de espermatozoides con cromatina dañada (r= -0.43104, p=0.0087), mientras que el porcentaje de motilidad se correlacionó negativamente con el porcentaje de espermatozoides con cromatina anormal (r= -0.48337, p=0.0421). En conclusión, el semen criopreservado de toros Brahman presenta un bajo nivel de espermatozoides con daño en la cromatina, lo cual se correlaciona negativamente con la motilidad y la vitalidad espermática.O objectivo deste estudo foi determinar a percentagem de espermatozóides com cromatina danificada, determinada pela coloração com azul de toluidina e sua relação com a viabilidade e a mobilidade do esperma cripreservado de touros Brahman. Para isso, foram utilizados três ejaculados de sêmen de seis touros Brahman, que uma vez descongelado foram coradas com azul de toluidina para determinar a integridade da cromatina (espermatozóides com cromatina normal coloream de azul ou verde; cromatina de espermatozóides con cromatina danificada, coloream de azul escuro ou violeta). Também foram corados com eosina nigrosina para determinar a viabilidade (espermatozóides vivos permanecem brancos e os mortos de cor rosa) e a motilidade espermática foi estimada por microscopia de luz. Foram encontradas diferenças significativas em todos os parâmetros, devido ao efeito de touro e o ejaculado, bem como a interacção entre essas duas variáveis. A percentagem de espermatozóides vivos foi de 50.02 ± 14.13% e motilidade espermática média de 33.88 ± 12.43%, enquanto a percentagem de espermatozóides com cromatina danificada foi de 4.17 ± 2.96%. A percentagem de espermatozóides vivos foi positivamente correlacionada com a motilidade (r=0.77217, p=0.0002) e negativamente com a porcentagem de espermatozóides com cromatina danificada (r = -0.43104, p= 0.0087), enquanto que a percentagem de motilidade correlacionou negativamente com a percentagem de espermatozóides com cromatina danificada (r = -0.48337, p=0.0421). Em conclusão, o sêmen de touros Brahman criopreservados tem um baixo nível de dano da cromatina, que está correlacionada negativamente com a motilidade e a vitalidade do esperma.

Background The aim of present study is to evaluate the effect of PolyVinyl Pyrrolidone (PVP) routinly used during ICSI procedure on sperm membrane integrity, and sperm chromatin status. Materials and methods This study was carried out on 21 semen samples from the infertile men referring to Isfahan Fertility and Infertility Center. The processed semen samples were divided into two portions. One portion was added to Ham’s F10+ 10% PVP, and the other portion was added to Ham’s F10 as a control group. Hypo-osmotic swelling test (HOST), SDS, and SDS+EDTA tests were carried out on the control and PVP groups at 15, 30, and 60min. Results The results show that sperm membrane integrity measured by HOST, and sperm chromatin stability measured by SDS test, reduces by increasing the exposure time to PVP. However, the ability of sperm chromatin undergoing decondensation (that has been assessed by SDS+EDTA), does not show any changes by increasing the exposure time to PVP. Conclusion The results of current study shows that reducing the exposure time of sperm to PVP may protect sperm membrane, and chromatin integrity.

Male germ cells undergo an extreme but fascinating process of chromatin remodeling that begins in the testis during the last phase of spermatogenesis and continues through epididymal sperm maturation. Most of the histones are replaced by small proteins named protamines, whose high basicity leads to a tight genomic compaction. This process is epigenetically regulated at many levels, not only by posttranslational modifications, but also by readers, writers, and erasers, in a context of a highly coordinated postmeiotic gene expression program. Protamines are key proteins for acquiring this highly specialized chromatin conformation, needed for sperm functionality. Interestingly, and contrary to what could be inferred from its very specific DNA-packaging function across protamine-containing species, human sperm chromatin contains a wide spectrum of protamine proteoforms, including truncated and posttranslationally modified proteoforms. The generation of protamine knock-out models revealed not only chromatin compaction defects, but also collateral sperm alterations contributing to infertile phenotypes, evidencing the importance of sperm chromatin protamination toward the generation of a new individual. The unique features of sperm chromatin have motivated its study, applying from conventional to the most ground-breaking techniques to disentangle its peculiarities and the cellular mechanisms governing its successful conferment, especially relevant from the protein point of view due to the important epigenetic role of sperm nuclear proteins. Gathering and contextualizing the most striking discoveries will provide a global understanding of the importance and complexity of achieving a proper chromatin compaction and exploring its implications on postfertilization events and beyond. This article is categorized under: Reproductive System Diseases > Genetics/Genomics/Epigenetics Reproductive System Diseases > Molecular and Cellular Physiology., (© 2022 The Authors. WIREs Mechanisms of Disease published by Wiley Periodicals LLC.)

Varicocele is one of the most common causes of primary male infertility. Pilea microphylla (PM) is being used as folk medicine. This study was aimed to investigate the effects of PM in a rat model of varicocele. A total of 30 male Wistar rats were divided into control, sham, varicocele, accessory varicocele and PM-treated groups. After 10 weeks of varicocele induction, sperm parameters and chromatin (Aniline blue, acridine orange and toluidine blue) were evaluated, except for the treated and accessory groups that received 50 mg/kg PM orally daily for 10 weeks and then were sacrificed. Sperm parameters significantly decreased in varicocele groups (P < 0.01). Moreover, there was a negative correlation between the DNA fragmentation and sperm parameters in varicocelized rats. Administration of PM led to significantly increased sperm parameters and AO staining (P < 0.05). These findings suggest that PM improves sperm parameters and DNA fragmentation in varicocelized rats. PM can reduce the damage to sperm DNA but not chromatin condensation.

There is evidence to show that varicocele repair can improve conventional sperm parameters but the effects on sperm chromatin integrity have not been fully elucidated. We sought to examine the effects of varicocelectomy on sperm maturation, nuclear chromatin integrity and nuclear sulfhydryl groups. We conducted a prospective study of consecutive infertile men (n=29) that underwent a microsurgical sub-inguinal varicocelectomy for treatment of a clinically palpable varicocele and abnormal semen parameters. Six healthy sperm donors served as controls. We evaluated conventional sperm parameters and markers of sperm chromatin and DNA integrity (aniline blue (AB) staining, iodoacetamide fluorescein (IAF) fluorescence and, % DNA fragmentation index (%DFI) and percent high DNA stainability (%HDS) by sperm chromatin structure assay) before and 6 months after surgery. The sperm %DFI, %HDS, % 5-IAF staining (diffuse head staining) and % AB staining (dark blue) were all significantly lower in the control group compared to infertile men with varicocele (8 vs. 20%, 4.0 vs. 9.6%, 1.7 vs. 16.3%, and 2.5 vs. 13.5% respectively). The %HDS and %DFI decreased significantly after surgery (from 10% to 6% and from 20% to 13%, respectively). Similarly, the %5-IAF and %AB staining also decreased significantly after surgery (from 16.3% to 5.4%, and from 13.5% to 5.4%, respectively). We observed significant inversely relationships between sperm progressive motility and both %IAF staining and %DFI (r=–0.44 and –0.43, respectively). The data show that varicocelectomy is associated with an improvement in sperm DNA integrity and chromatin compaction using three different assays of sperm chromatin integrity. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate sperm chromatin maturity and integrity of that injected into good-quality oocytes in anin vitrofertilization-intra cytoplasmic sperm injection (IVF-ICSI) program. A cut-off value of sperm chromatin maturity and integrity was developed as a function of their correlation to the zygote development, i.e., embryo formation and cleavage rate. The study assessed sperm chromatin maturity using aniline blue (AB) staining, whereas toluidine blue (TB) staining was used to assess sperm chromatin integrity. Ejaculates from 59 patients undergoing ICSI and 46 fertile normozoospermic donors for determination of normal values of sperm chromatin status were used in this study. Embryo formation and cleavage rates were observed for the period of 3 days after ICSI. There was a significant difference in the percentage of sperm with mature chromatin between ejaculate from ICSI patients and fertile donor (p=0.020); while there was no significant difference in sperm chromatin integrity of both samples (p=0.120). There was no significant correlation between sperm chromatin maturity and either embryo formation or cleavage rate; as well as sperm chromatin integrity to both parameters of zygote development (p>0.05). Furthermore, we found that the cut-off value of sperm chromatin maturity and integrity of the fertile normozoospermic ejaculates were 87.2% and 80.2%, respectively. Using the cut-offs, we found that low sperm chromatin maturity at the level of <87% correlated significantly with the cleavage rate of the zygote (p=0.022; r=0.371); whereas poor sperm chromatin integrity at the level of <80% correlated with embryo formation (p=0.048; r=0,485). In conclusion, this study showed that poor maturity and integrity of sperm chromatin (AB<87% and TB<80%, respectively), could affect zygote development following ICSI. Abbreviations: AB: aniline blue; CMA3: chromomycin A3; ICSI: intra cytoplasmic sperm injection; IVF:in vitrofertilization; PBS: phosphate buffer saline; SPSS: Statistical Package for Social Science; TB: toluidine blue; WHO: World Health Organization [ABSTRACT FROM PUBLISHER]

This study is aimed at evaluating the association between occupational exposure to organophosphate (OP) and carbamate (CB) pesticides and semen quality as well as levels of reproductive and thyroid hormones of pesticide sprayers in Malihabad, Lucknow, Uttar Pradesh, India. Thirty-five healthy men (unexposed group) and 64 male pesticide sprayers (exposed group) were recruited for clinical evaluation of fertility status. Fresh semen samples were evaluated for sperm quality and analyzed for DNA fragmentation index (DFI) by flow cytometry. Pesticide exposure was assessed by measuring erythrocyte acetylcholinesterase and plasma butyrylcholinesterase (BuChE) with a Test-mate ChE field kit. Serum levels of total testosterone (Tt), prolactin (PRL), follicle-stimulating hormone (FSH), luteinizing hormone (LH), thyroid-stimulating hormone (TSH), and free thyroxine (FT4) were analyzed using enzyme immunoassay kits. Evidence of pesticide exposure was found in 88.5% of sprayers and significant increments were observed in sperm DFI with significant decrease in some semen parameters. DFI was negatively correlated with BuChE, sperm concentration, morphology, and vitality in these pesticide sprayers. The levels of Tt, PRL, FT4, and TSH appeared to be normal; however, there was a tendency for increased LH and FSH levels in exposed workers. The results confirm the potential impact of chronic occupational exposure to OP and CB pesticides on male reproductive function, which may cause damage to sperm chromatin, decrease semen quality, and produce alterations in reproductive hormones, leading to adverse reproductive health outcomes. [ABSTRACT FROM AUTHOR]

Aim of the study The purpose of this study was to evaluate the dynamic of the sperm DNA integrity of spermatozoa from endangered Grey Steppe cattle breed bulls in order to establish the potential use of sperm DNA fragmentation (SDF) assay for improving the routine fertility screening in the conservation program. Aim of the study The purpose of this study was to evaluate the dynamic of the sperm DNA integrity of spermatozoa from endangered Grey Steppe cattle breed bulls in order to establish the potential use of sperm DNA fragmentation (SDF) assay for improving the routine fertility screening in the conservation program. Results Variability of percentages of spermatozoa with fragmented DNA was assessed in 2 Grey Steppe bulls. In the case of both bulls, the sperm DNA fragmentation index (DFI%) increased progresively during the 6 hours incubation, when the hightest DNA fragmentation index (DFI) was observed, 7.5% in bull 2, respectively, 4.5% in bull 1. At T0, the DFI% was greater for Bull 2, a value of 3.5% being registered as compared to bull 1, in which the DFI% was 2.5%. Conclusions The semen from the two Grey Steppe bulls analyzed in this study presented moderate DNA fragmentation index and may be used for AI and IVF in conservation program of the Grey Steppe breed. [ABSTRACT FROM AUTHOR]

A strategy to circumvent the poor polyphenols bioavailability is to load these compounds into liposomes. We evaluated the in vitro effects of quercetin (Q) and Q-loaded liposomes (QLL, 30, 50, 100 μM) on motility, viability and chromatin integrity of swim-up selected human sperm. Antioxidant power was assayed against tert-butylhydroperoxide induced lipid peroxidation (LPO) using C11-BODIPY581/591 fluorescent probe and transmission electron microscopy. QLL showed decreased toxicity for sperm motility and viability and increased DNA damage compared to Q. The percentage of sperm with fluorescence, marker of LPO, was decreased in samples incubated with Q vs QLL (P < 0.001). The ultrastructure of acrosomes and membranes was preserved with Q 30/100 μM, whereas QLL did not prevent membrane injury. Q alone appeared more effective than Q incorporated into liposomes; however liposomes could be considered as carriers that may convey different compounds inside sperm; they may therefore represent a field of research rich of many applications. [ABSTRACT FROM AUTHOR]

Moringa oleifera L. is a medicinal plant with potential antioxidant property. This study was aimed at investigating the chemoprotective effect of Moringa oleifera leaf extract ( MOE) on cyclophosphamide ( CP)-induced testicular toxicity. Two-week-old male Swiss albino mice were intraperitoneally injected with phosphate-buffered saline, 50 mg kg−1 of CP and 25 mg kg−1 of MOE. In combination treatment, mice were injected with 25 mg kg−1 of MOE 24 h prior to CP injection, 24 h prior and post- CP injection and 24 h post- CP injection for 5 consecutive days (10 mg kg−1). Six weeks later, mice were sacrificed to assess epididymal sperm parameters. MOE alone did not have any significant effect on sperm parameters. However, acute injection of CP resulted in significant decline in motility ( P < 0.001), increase in head abnormality ( P < 0.01) and DNA damage ( P < 0.05). Combining MOE with CP increased the sperm density, motility and reduced head defect and DNA damage, irrespective of the schedule and dosage of MOE. Administration of MOE prior to CP significantly elevated the level of superoxide dismutase and catalase with concomitant decrease in lipid peroxidation in the testicular tissue. In conclusion, MOE may have potential benefit in reducing the loss of male gonadal function following chemotherapy. [ABSTRACT FROM AUTHOR]

Copyright of Revista Cientifica de la Facultade de Veterinaria is the property of Universidad del Zulia, Facultad de Ciencias Veterinarias and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Summary The formation of motile spermatozoa involves the highly conserved formation of protamine-rich, tightly packed chromatin. However, genetic loss of protamine function in Drosophila and mice does not lead to significant decompaction of sperm chromatin. This indicates that other proteins act redundantly or together with protamines. Here, we identify Prtl99C as a Drosophila sperm chromatin-associated protein that is essential for male fertility. Whereas the loss of protamines results in modest elongation of sperm nuclei, knockdown of Prtl99C has a much stronger effect on sperm nuclei. Loss of protamines and Prtl99C indicates an additive effect of these proteins on chromatin compaction, in agreement with independent loading of these factors into sperm chromatin. These data reveal that at least three chromatin-binding proteins act together in chromatin reorganization to compact the paternal chromatin. [ABSTRACT FROM AUTHOR]

Study Question: Do the luminal fluids of the epididymis and the vas deferens contribute to sperm chromatin fragmentation (SCF) in mice?Summary Answer: The luminal fluids of both organs are required for activating SCF in mice, but the vas deferens luminal fluid does this more efficiently than that of the epididymis.What Is Known Already: Mice sperm have the ability to degrade their DNA in an apoptotic-like fashion when treated with divalent cations in a process termed SCF. SCF has two steps: the induction of reversible double-strand DNA breaks at the nuclear matrix attachment sites, followed by the irreversible degradation of DNA by nuclease. Single stranded DNA breaks accompany SCF.Study Design, Size, Duration: Luminal fluids from two reproductive organs of the mouse (B6D2F1 strain), the epididymis and vas deferens, were extracted and tested for SCF activation with divalent cations using four different combinations of the sperm and the surrounding luminal fluids: (i) in situ--sperm were kept in their luminal fluid and activated directly; (ii) reconstituted--sperm were centrifuged and resuspended in their luminal fluid before SCF activation; (iii) mixed--sperm were centrifuged and resuspended in the luminal fluid of the other organ; (iv) no luminal fluid--sperm were centrifuged and reconstituted in buffer. All four experiments were performed without (controls) and with divalent cations (resulting in SCF). For each experimental condition, two different mice were used and the analyses averaged.Participants/materials, Setting, Methods: DNA damage by SCF was analyzed by three different methods, the sperm chromatin structure assay (SCSA), terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) analysis and field inversion gel electrophoresis.Main Results and the Role Of Chance: In all three assays that we used, the vas deferens luminal fluid was much more efficient in stimulating SCF in the sperm from either source than that of the epididymis (P < 0.0001). Vas deferens sperm were capable of initiating lower levels of SCF in the absence of luminal fluid (P < 0.0001).Limitations, Reasons For Caution: Analyses were performed in only one species, the mouse, but we used three separate assays in our analysis.Wider Implications Of the Findings: The data suggest that the luminal fluid of the male reproductive tract interacts with sperm during their transit providing a mechanism to degrade the DNA. We hypothesize that this is part of an apoptotic-like mechanism that allows the reproductive tract to eliminate defective sperm. The SCF model also allowed us to identify differences in the types of DNA lesions that the three tests can identify, providing important background information for the use of these tests clinically. [ABSTRACT FROM AUTHOR]

Introduction: Different studies have shown that there is no relation between sperm chromatin stability and fertilization rate in both IVF and ICSI patients. However, the relation between SDS tests, as a detergent, along with DTT as reducer of disulphide bridges has not been studied so far in ICSI patients. Since different concentrations of DTT can induce different degrees of sperm chromatin decondensation, the aim of this study was to evaluate the effect of different concentrations of DTT on sperm chromatin decondensation in IVF and ICSI cases. Methods: During this study, 85 patients were divided into two groups according to their treatment procedure (IVF or ICSI).Semen samples of each patient was evaluated for sperm chromatin tests including SDS, SDS+EDTA & SDS+DTT for assessment of free thiole groups level (-SH), amount of non covalent bond between Zn and thioles(-SH Zn SH-) and levels of disulfide bond (-S-S-) in sperm chromatin, respectively. In this study, seminal fructose concentration, corrected seminal fructose level and true corrected fructose level as indicators of seminal vesicle function on sperm chromatin stability were assessed. Results: No correlation was observed between any of the above tests and rate of fertilization, both in IVF and ICSI cases. However, in IVF patients, a significant correlation was observed between SDS, SDS+DTT test and seminal fructose level, while in ICSI patients, only a significant correlation was observed between SDS+DTT and corrected or true fructose concentration. Conclusion: Since no correlation was observed between sperm chromatin test and fertilization rate, it is suggested that the chromatin status of these samples are adequate for fertilization to take place and extent of disulphide bridges has no effect on fertilization rate. However, the amount of disulphide bound present in sperms of ICSI and IVF patients are different, and this difference is related to seminal vesicle performance in these patients.

Varicocele is one of the most common causes of primary male infertility. Pilea microphylla (PM) is being used as folk medicine. This study was aimed to investigate the effects of PM in a rat model of varicocele. A total of 30 male Wistar rats were divided into control, sham, varicocele, accessory varicocele and PM-treated groups. After 10 weeks of varicocele induction, sperm parameters and chromatin (Aniline blue, acridine orange and toluidine blue) were evaluated, except for the treated and accessory groups that received 50 mg/kg PM orally daily for 10 weeks and then were sacrificed. Sperm parameters significantly decreased in varicocele groups (P < 0.01). Moreover, there was a negative correlation between the DNA fragmentation and sperm parameters in varicocelized rats. Administration of PM led to significantly increased sperm parameters and AO staining (P < 0.05). These findings suggest that PM improves sperm parameters and DNA fragmentation in varicocelized rats. PM can reduce the damage to sperm DNA but not chromatin condensation. [ABSTRACT FROM AUTHOR]

The classical idea about the function of the mammalian sperm chromatin is that it serves to transmit a highly protected and transcriptionally inactive paternal genome, largely condensed by protamines, to the next generation. In addition, recent sperm chromatin genome-wide dissection studies indicate the presence of a differential distribution of the genes and repetitive sequences in the protamine-condensed and histone-condensed sperm chromatin domains, which could be potentially involved in regulatory roles after fertilization. Interestingly, recent proteomic studies have shown that sperm chromatin contains many additional proteins, in addition to the abundant histones and protamines, with specific modifications and chromatin affinity features which are also delivered to the oocyte. Both gene and protein signatures seem to be altered in infertile patients and, as such, are consistent with the potential involvement of the sperm chromatin landscape in early embryo development. This present work reviews the available information on the composition of the human sperm chromatin and its epigenetic potential, with a particular focus on recent results derived from high-throughput genomic and proteomic studies. As a complement, we provide experimental evidence for the detection of phosphorylations and acetylations in human protamine 1 using a mass spectrometry approach. The available data indicate that the sperm chromatin is much more complex than what it was previously thought, raising the possibility that it could also serve to transmit crucial paternal epigenetic information to the embryo. [ABSTRACT FROM AUTHOR]

Scientific concern exists about the toxic effect of dichlorodiphenyldichloroethylene ( p, p′-DDE) and lindane on male infertility, and the mechanism underlying male reproductive toxicity of this pesticide remains unanswered. We investigated not only the possible association between the chlorinated pesticide levels and semen quality in nonoccupationally exposed men, but also the probable mode of action using mitochondrial membrane potential (MMP), reactive oxygen species (ROS), lipid peroxidation (LPO), and sperm chromatin structure assay (SCSA). A study in 278 men (21-40 years old) who visited Obstetrics and Gynecology Department, KGMU, Lucknow, for semen analysis was conducted. We performed semen analysis according to the WHO guidelines, while p, p′-DDE and lindane analysis was done by the GLC and LPO by the spectrophotometer, and the sperm mitochondrial status, ROS, and SCSA with the flow cytometer. The questionnaire data showed no significant difference in the demographic characteristics between the two groups, i.e., trying to conceive >1 year and proven fertility. However, a significant difference in the concentration of p, p′-DDE and lindane was observed between the groups. When the subjects were divided among four categories by quartile of exposure, the subjects in the highest quartile showed low sperm motility as compared to the subjects in the lowest quartile. Pearson's correlation showed a significant negative correlation between semen p, p′-DDE, lindane level, and sperm quality and positive association with the number of cells with depolarized mitochondria, elevation in ROS production and LPO, and DNA fragmentation index (DFI). The findings are suggestive that these toxicants might cause a decline in semen quality, and these effects might be ROS, LPO, and mitochondrial dysfunction mediated. [ABSTRACT FROM AUTHOR]

Background: Currently, relation between reactive oxygen species (ROS) ROS concentration and semen quality was indicated. Saffron has traditionally been not only considered as a food additive but also as a medicinal herb, which has a good antioxidant properties. Objective: The aim of this study was to evaluate the protection potency of saffron and vitamin E on sperm chromatin integrity. Materials and Methods: Thirty adult male Wistar rats divided equally into saffron (100 mg/kg), vitamin E (100 mg/kg) and control (0.5cc distilled water /day) groups. After 60 days, cauda epididymis dissected and sperm cells were used for analysis of sperm chromatin packaging by chromomycin A3 (CMA3) staining, and sperm chromatin susceptibility to acid denaturation by acridine orange (AO) staining Results: The mean percentage of CMA3 positive sperm was significantly decreased in saffron and vitamin E groups relative to control group (p<0.001). Moreover, the AO staining results showed that the mean percentage of sperm with DNA damage was significantly decreased in saffron and vitamin E groups as compared with control group (p<0.001). Conclusion: Our results purposed that saffron can protect sperm against DNA damage and chromatin anomalies. [ABSTRACT FROM AUTHOR]

Background: Sperm DNA damage is associated with male infertility but whether normozoospermic infertile men also have DNA damage is unknown. Objective: To evaluate sperm DNA and chromatin integrity in men with mild male factor infertility. Design, setting and participants: Prospective study of 102 consecutive men (78 normozoospermic, 15 asthenozoospermic, 9 oligozoospermic) enrolled for intrauterine insemination (IUI) and 15 fertile controls. Outcome measurements and statistical analysis: Standard semen parameters and sperm chromatin and DNA integrity were assessed and compared between groups. Sperm chromatin quality was assessed by (1) aniline blue staining (AB is specific to histone lysines), (2) iodoacetamide fluorescein fluorescence (IAF targets free protamine sulfhydryl groups) and (3) sperm chromatin structure assay (SCSA) with the results expressed as % DNA fragmentation index (%DFI). Results and limitations: The mean (±SD) percentage of spermatozoa with positive IAF fluorescence was significantly higher in the IUI population compared to fertile controls (17 % ± 10 % vs. 8 % ± 6 %, P = 0.0011) and also in the normozoospermic subset ( n = 78) compared to controls (16 % ± 9 % vs. 8 % ± 6 %, P < 0.0001, ANOVA). We also observed a trend toward lower %progressive motility, and higher %AB staining and %DFI in the IUI group compared to controls. We observed significant relationships between sperm %DFI and progressive motility ( r = −0.40, P < 0.0001) and between positive AB staining and IAF fluorescence ( r = 0.58, P < 0.0001). Conclusions: The data indicate that sperm chromatin integrity may be abnormal in men enrolled in IUI treatment cycles, despite the fact that most of these men are normozoospermic. [ABSTRACT FROM AUTHOR]

Abstract: The objective of the present study was to explore the potential relationships of ovine sperm chromatin integrity, quantified using the sperm chromatin structure assay (SCSA), to the heat load of the scrotum and the discomfort felt by the animals because of fluctuations of microclimatic factors at different time periods before ejaculation. Ejaculates were collected once per week from five Chios rams and four East Friesian rams for 12 months and stored in liquid nitrogen. Frozen-thawed semen samples were analyzed using the SCSA, to determine the DNA fragmentation index (DFI) and the percentage of cells outside the main sperm population (%DFI) in each one of the samples. Scrotal surface temperature (SST) of each ram was measured using an infrared thermometer on a daily basis. Ambient air temperature and relative humidity were recorded at hourly intervals throughout the experimental period and temperature-humidity index (THI) was used to assess the discomfort felt by the rams. Mean values of SST (SSTmean) and THI (THImean) were computed for eight different time periods (up to 61 days) preceding each ejaculation day (Day 0). A linear mixed-effect model analysis was performed to describe the relation of SCSA parameters to collection month, SSTmean, and THImean of different time periods before ejaculation. The results of the statistical analysis revealed a relation of %DFI to the SSTmean of the last 12 days preceding ejaculation, namely the period that resembled the phase of epididymal maturation. On the contrary, the variation of DFI was most adequately described by the linear mixed-effect model applied for Days 54 to 48 before ejaculation, which resembled the phase of spermatogonial mitoses. The effect of collection month was significant for DFI and %DFI, with semen samples collected in September and February exhibiting the lowest DFI values; a less profound seasonal pattern was detected for %DFI. The effect of THImean on DFI and %DFI was proven nonsignificant in regard to all time periods. In conclusion, a relation of SCSA parameters to SSTmean of different periods before ejaculation was shown in the present study, implying an effect of scrotal microenvironment on intratesticular and epididymal sperm population. In contrast, we failed to detect any effect of microclimate-induced discomfort felt by the animals on the chromatin integrity of frozen-thawed ram spermatozoa. [Copyright &y& Elsevier]

Sperm chromatin integrity may affect the outcomes of assisted reproductive technology ( ART). Developing a clinically reliable strategy to enrich sperm samples with high chromatin quality spermatozoa prior to sperm banking or use in ART would thus be advantageous. The objectives of this study were to: (i) assess the sperm chromatin quality in men with different categories of semen parameters; and (ii) evaluate the extents of Annexin-V magnetic-activated cell sorting ( MACS) technology coupled with differential density gradient centrifugation ( DGC) in improving sperm chromatin quality. Three categories of men from couples attending a university-based fertility clinic were recruited based on their semen parameters: normozoospermic ( n = 13), asthenoteratozoospermic ( n = 17) and teratozoospermic ( n = 12). For each patient, spermatozoa in semen samples were processed first by DGC to enrich the motility and further by MACS to remove spermatozoa showing apoptotic features. The yield and enrichment of sperm quality was evaluated at each step with conventional semen parameters in conjunction with a combination of five complementary assays, to assess sperm maturity, chromatin structure, compaction and DNA integrity (Hyaluronic Binding Assay, SCSA, chromomycine A3 staining and TUNEL and COMET assays). Our results demonstrated that, compared with normozoospermic samples, raw asthenoteratozoospermic and teratozoospermic samples had a higher proportion of spermatozoa containing DNA breaks, but only asthenoteratozoospermic exhibited altered chromatin structure and decreased binding to hyaluronic acid. Interestingly, the DGC appeared to select for more mature spermatozoa with high DNA compaction. More importantly, in all categories of semen samples, Annexin-V MACS allows enrichment of spermatozoa with good chromatin quality as measured by the TUNEL and SCSA. Because effective treatment modalities to improve sperm DNA damage are limited, our results suggest a potential clinical value of MACS as a mean to enhance sperm quality that may improve assisted reproductive outcomes. [ABSTRACT FROM AUTHOR]

Abstract: The objective of this study was to improve the quality of cryopreserved–thawed equine sperm using single-layer density centrifugation (SLC). Sperm quality was assessed by DNA integrity, motility, morphology, mitochondrial membrane potential, viability, and plasma membrane alteration. The percentage of DNA-damaged sperm (expressed in % COMP) was lower (P = .001) after SLC (1.6 ± 0.5% vs. 6.8 ± 0.5%). Total sperm motility (80 ± 2.4% vs. 41.7 ± 2.4%) and progressive sperm motility (69.5 ± 2.9% vs. 31.5 ± 2.9%) (P < .001), as well as the percentage of morphologically normal sperm (45 ± 3.9% vs. 27.7 ± 3.9%), increased after SLC compared with control sample. In addition, the proportion of sperm with high mitochondrial membrane potential increased (81.6 ± 1.8% vs. 42.1 ± 1.8%), as did the viability of sperm (71.1 ± 2.4% vs. 39.5 ± 2.4%), after SLC compared with the control sperm. The proportion of sperm with alteration in plasma membrane structure was lower after SLC compared with control sample (6.4 ± 1.1% vs. 18.9 ± 1.1%). Overall, sperm recovery was 72.7 ± 3.6% in the control sample compared with 14.8 ± 3.6% after SLC (P < .001). We conclude that based on the sperm parameters evaluated, SLC improves the quality of cryopreserved–thawed equine spermatozoa. [Copyright &y& Elsevier]

Abstract: Sperm chromatin status was assessed in 565 Zebu and Zebu crossbred beef bulls in extensive tropical environments using the sperm chromatin structure assay (SCSA). The SCSA involved exposure of sperm to acid hydrolysis for 0.5 or 5.0 minutes, followed by flow cytometry to ascertain relative amounts of double-stranded (normal) and single-stranded (denatured) DNA, which was used to generate a DNA fragmentation index (%DFI). With conventional SCSA (0.5-minute SCSA), 513 bulls (91%) had <15 %DFI, 24 bulls (4%) had 15 to 27 %DFI, and 28 bulls (5%) had >27 %DFI. In 5.0-minute SCSA, 432 bulls (76%) had <15 %DFI, 68 bulls (12%) had 15 to 27 %DFI and 65 bulls (12%) had >27 %DFI. For most bulls, the SCSA was repeatable on two to four occasions; however, because most bulls had <15 %DFI, repeatability of the SCSA will need to be determined in a larger number of bulls in the 15 to 27 %DFI and >27 %DFI categories. The %DFI was negatively correlated with several bull semen parameters and the strongest negative correlation was with normal sperm. There was a strong positive correlation between %DFI and sperm head abnormalities. Based on these findings, most Zebu beef bulls in extensive tropical environments had relatively stable sperm chromatin. Based on the apparent negative correlations with conventional semen parameters, we inferred that the SCSA measured a unique feature of sperm quality, which has also been suggested for other species. Further studies on the relationships between sperm chromatin stability and fertility are required in beef bulls before chromatin status can be used as an additional predictor of the siring capacity of individual bulls in extensive multiple-sire herds. [Copyright &y& Elsevier]

Background and Objective: Protamine deficiency and DNA fragmentation have a profound effect on embryo gene expression. Incubation of sperm in culture medium leads to morphological changes of the nucleus and DNA release. However, the relationship between protamine deficiency and DNA fragments released from incubated sperm and the presence of the DNMT1 gene is not known. Materials and Methods: Semen analysis was performed according to the WHO criteria. CMA3 staining was used to determine protamine deficiency. Twenty million sperms per sample were incubated in Ham's F10 medium for 24 hr at 37 °C. After centrifugation, DNA was extracted from the supernatant. DNA concentration was measured by biophotometer. The presence of the DNMT1 gene was confirmed by PCR using specific primers and agarose gel electrophoresis. Results: A significant correlation was observed between the DNA fragments released from sperm in culture media and CMA3 staining (P<0.05). The presence of DNMT1 gene was significantly correlated with DNA fragments released from the sperm (P<0.05). Protamine deficiency and the presence of DNMT1 gene were not significantly correlated. Conclusion: These results demonstrate that incubation of protamines-deficient sperm leads to the release of additional DNA fragments that eventually could have detrimental effectson embryo’s health. [ABSTRACT FROM AUTHOR]

Background: Diabetes mellitus (DM), primary or idiopathic is a chronic disorder of the carbohydrate, lipid and protein metabolism. DM may impact male reproductive function at several levels. It is shown that DM has detrimental effects on sperm parameters in human and experimental animals. Objective: The aim of this study was to observe the effects of diabetes on sperm parameters (viability, count, morphology and motility) and evaluation of sperm chromatin quality in mice. Materials and Methods: Totally twenty adult male Syrian mice were divided randomly into 2 groups (n=10). The animals of group A were considered as controls while group B mice were diabetic that received a single dose (200 mg/kg) streptozotocin (STZ) intra peritoneally. After 35 days, the cauda epididymis of each diabetic mouse was dissected and placed in culture medium for 30 min. The swim-out spermatozoa were analyzed for count, motility, morphology and viability. The sperm chromatin quality and DNA integrity, was evaluated with Aniline Blue (AB), Toluidine blue (TB), Acridine orange (AO) and Chromomycin A3 (CMA3) staining. Results: In sperm analysis, the diabetic mice had poor parameters in comparison with control animals (p=0.000). Regarding sperm chromatin quality, the results of TB and AO tests showed statically significant differences between two groups, but in AB and CMA3 staining, we didn't see any differences between them. Conclusion: The results showed that STZ-induced diabetes mellitus may influence the male fertility potential via affecting sperm parameters and DNA integrity in mice. However, according to our data, the diabetes doesn't have any detrimental effects on histone-protamines replacement during the testicular phase of sperm chromatin packaging. [ABSTRACT FROM AUTHOR]

BACKGROUND: Follicular fluid (FF) is biological fluid rich in nutrients, growth factors, hormones and may affect the sperm quality. Sperm washing has been done using conventional media in laboratory procedure so far. AIM: This study aimed to investigate the effects of FF on survival and maintenance of chromatin integrity post swim up. MATERIALS AND METHODS: Each washed semen sample was divided into two parts; the control group was incubated in the media, and the experimental groups incubated in the media containing 10% follicular fluid. Smears were prepared after 20 min, 180 min, 24 hours and no incubation times. Sperm chromatin changes like protamine, histone, DNA denaturation, sperm chromatin stability and motility were evaluated at different times. RESULTS: Incubation of sperm in the follicular fluid increased sperms with normal histone, normal chromatin protamine and sperm with normal head size (p < 0.05). CONCLUSIONS: Administration of follicular fluid into the culture media of the sperms that had been separated by swim up method could improve the sperm quality. Further studies are recommended for understanding the mechanism of the structural change of the sperm chromatin. [ABSTRACT FROM AUTHOR]

Background: It has been claimed that by using different washing methods, the sperms can be separated according to size, motility, density, chromosomal content and surface markings and charge. These methods also reduce sperm chromatin deficiencies and screen the sperms before applying in assisted reproduction techniques. Objective: This study compared simple density gradient methods and a combined method with albumin density gradient and PureSperm separation (alb/PureSperm) for sex preselection by double fluorescence in situ hybridization (FISH) versus chromomycin A3 staining to determine chromatin integrity. Materials and Methods: 30 normal semen samples were prepared with PureSperm, albumin gradients and alb/PureSperm. All samples were then stained by FISH and chromomycin A3. The results were compared with SPSS 11.5 and the Kruskal- Wallis test. Results: The proportion of X-bearing spermatozoa by PureSperm separation (47.58±5.67) and Y-bearing spermatozoa by albumin gradient (46.13±3.83) methods were slightly higher than in putative normal sperm samples (1:1), but there were no significant differences in the X- or Y- bearing spermatozoa counts among the three methods. Albumin gradient separation tended to underestimate abnormal spermatozoa compared to PureSperm and combined alb/PureSperm. Conclusion: Routine separation methods slightly enriched X- or Y- bearing spermatozoa, but the differences were not significant for clinical purposes. The combined alb/PureSperm method had no advantages for assessing sex ratio or chromatin integrity compared to simpler gradient methods. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of a mild induced testicular and epididymal hyperthermia (+2°C) on sperm chromatin integrity in men. Design: Experimental prospective study. Setting: University hospital. Patient(s): Five healthy fertile volunteers. Intervention(s): Testicular and epididymal hyperthermia was induced by maintaining the testes at inguinal position with the support of specially designed underwear 15 ± 1 hours daily for 120 consecutive days. Main Outcome Measure(s): Classic semen characteristics. Sperm DNA fragmentation index (DFI) and high DNA stainability (HDS) were analyzed by sperm chromatin structure assay. Result(s): Compared with baseline values, sperm DFI and HDS were significantly increased as early as day (D) 20 and D34, respectively, and remained elevated during the entire period of hyperthermia. Percentages of motile and viable spermatozoa decreased as early as D20 and D34, respectively, and total sperm count decreased at D34 during hyperthermia and remained low during the entire hyperthermia period. All studied parameters returned to respective baseline values at D73 after cessation of hyperthermia. Conclusion(s): Mild induced testicular and epididymal hyperthermia largely impaired sperm chromatin integrity, which appeared before any changes in sperm output. These findings may have clinical implications in male contraception, infertility, and assisted reproductive technology. [ABSTRACT FROM AUTHOR]

Background and purpose: The sperm of infertile men with varicocele exhibit markedly high DNA damage that appears to be related to high oxidative stress (OS). Aminoguanidine (AG) is a specific inhibitor of the nitric oxide synthase (NOS) isoforms iNOS and an antioxidant, the effects of which decrease NO and peroxynitrite production. The aim of this study was to determine the effects of AG on sperm chromatin in varicocelized rats. Methods: Thirty male Wistar rats were divided into 5 groups: control, sham, varicocele, and AG and placebo-treated groups. At 10 weeks after varicocele induction, sperm chromatin was evaluated in all groups, except in the treated groups. The treated groups received intraperitoneal injections of 50 mg/kg AG or placebo daily for 10 weeks and then were killed for chromatin assessment. Sperm chromatin was evaluated by aniline blue, acridine orange, toluidine blue, and chromomycin A3 staining. Results: The results of the 4 above tests were significantly increased between varicocele and control (and sham) groups (P < .05). Conclusion: The findings of this study suggest that AG improves sperm DNA fragmentation that is associated with infertility in varicocelized rats, and treatment with AG can reduce the damage to sperm DNA. [ABSTRACT FROM PUBLISHER]