bluuebunny/crossref_metadata_2025 huggingface dataset:
Diel Pattern of Photosynthate Partitioning in Phytoplankton Population in an Oligomesotrophic Lake
Authors: Maurin Nadine , Amblard Christian , Bourdier Gilles
Published: October 2023

The aim of this study was to measure the short‐term changes in inorganic carbon allocation into various macromolecular compounds (proteins, polysaccharides and lipids) throughout a diurnal cycle in the oligomesotrophic Lake Pavin (Massif Central of France) at the depths of 5, 15 and 30 m. Biochemical fractionation was done by consecutive differential extractions in order to separate proteins, polysaccharides, lipids and low molecular weight compounds (LMW) by virtue of their relative solubilities in different extraction solvents. Over the entire diurnal cycle inorganic carbon was preferentially incorporated into proteins (M = 30%), then into polysaccharides (M = 28%), LMWs (M = 27%) and lipids (M = 15%). However, at 5 m, diurnal variations were reflected by the high percentage of the inorganic carbon incorporated into polysaccharides during periods of high light intensity and decreased at dawn and dusk. The reverse pattern was observed for the allocation of inorganic carbon to proteins.
Distribution Patterns, Species Assemblages and Habitat Selection of the Stoneflies (Plecoptera) from Two Mediterranean River Basins in Southern Spain
Authors: Prenda José , Gallardo‐Mayenco Alfonso
Published: October 2023

The stonefly distribution and its relationship with habitat changes, based on stream physico chemical features, were investigated in two Mediterranean river basins (Guadaira and Guadalete, S Spain). Plecopteran nymph distribution varied along an elevation gradient. Both stonefly species richness and species diversity increased with altitude. Stoneflies were able to tolerate a wide range of environmental conditions but were absent from low altitude sites, temporary streams with high salinity values and high water temperature. A total of 13 stonefly species were found in the study area, from which Tyrrhenoleuctra minuta (Klapálek) and Protonemura sp. were dominant. The most widespread species were Nemoura lacustris and Capnioneura mitis. Three distinct species assemblages were observed according to their distribution patterns, but with similar environmental preferences. Capnioneura petitpierreae and T. minuta showed preferences to low altitude sites, whereas C. mitis and N. lacustris were found throughout all sites. T. minuta and N. lacustris inhabited permanent and temporary sites, while the same species and C. mitis also were found in semipermanent streams. Temporally, most stoneflies were caught during winter (85.9%), especially at the low altitude sites.
Effects of Food Availability and Predation on a Marine Zooplankton Community — a Study on Copepods in the Baltic Sea
Authors: Adrian Rita , Hansson Sture , Sandin Barbro , Stasio De Bart , Larsson Ulf
Published: October 2023

The importance of food resources versus predatory effects in determining the abundance pattern of zooplankton varies over time and space. Here, we evaluate the population dynamics of dominant calanoid copepods (Acartia spp. bifilosa/A. longiremis and Eurytemora affinis) in a reference area and a eutrophied area in the northern Baltic Sea Proper. We base the paper on a combination of statistical analyses of a long term (12 years) data set on population biomass dynamics, laboratory experiments on the feeding biology of these species and literature data of their susceptibility to predation. Overall, copepods were equally abundant at both sites, but Eurytemora was more abundant in the eutrophied area while Acartia spp. dominated in the non‐eutrophied reference area. There was, however, no significant difference between the two areas in biomass dynamics (biomass specific rate of change) of the two groups of copepods. However, in spring and early summer the population biomass increased faster for Eurytemora as compared to Acartia spp. This is consistent with our findings from feeding and respiration experiments. Eurytemora exhibited generally higher ingestion rates and had a higher growth efficiency (ratio of ingestion to respiration rates), which suggests an advantage over Acartia spp. when food availability increases. Both species generally showed a rapid population decline starting in late summer, most likely caused by intensive predation by fish and mysid shrimps. In late autumn and winter, biomasses of both genera continued to decrease, but the decline was fastest for Eurytemora. During this period of the year, when the food availability is sparse, Acartia spp. may have an advantage over Eurytemora, as we found that the former ingests a broader size range of food particles and it has also been reported to be able to switch from filter feeding to raptorial feeding. This study shows that dominant copepods in the net‐zooplankton community in our study area were structured both by food availability and predation. The relative importance of these effects varied seasonally.
Effects of Accessory Sex Gland Fluid on Viability, Capacitation, and the Acrosome Reaction of Cauda Epididymal Bull Spermatozoa
Authors: WAY AMY L. , GRIEL LESTER C. , KILLIAN GARY J.
Published: October 2023

ABSTRACT: The effect of accessory sex gland fluid (AGF) on viability and acrosomal integrity of spermatozoa was examined with cauda epididymal spermatozoa and AGF from the same Holstein bull (n = 6). Surgical cannulation of the vasa deferentia enabled the separate collection of cauda epididymal effluent and AGF from each bull. Cauda epididymal effluent was incubated with either AGF collected from the same bull or medium alone. Following coincubation, spermatozoa (5 × 107 sperm/mL) were incubated in medium alone or under capacitating conditions (10 μg/mL heparin) for 16 hours. Every 2 hours, an aliquot of spermatozoa was exposed to lysophosphatidylcholine (121 μg/mL) to induce the acrosome reaction in capacitated spermatozoa. Sperm motility decreased over time re gardless of treatment. Overall, spermatozoa incubated in AGF had fewer acrosome‐intact live spermatozoa than did those not incubated in AGF. Viability was significantly (P < .05) compromised over time when spermatozoa were exposed to AGF, compared with those not preincubated in AGF. Significantly more (P < .05) acrosome‐reacted live spermatozoa were seen following exposure to heparin and lysophosphatidylcholine when spermatozoa were not preincubated in AGF. We conclude that exposure of spermatozoa to AGF accelerates cell death and that rapid removal of spermatozoa from seminal plasma is critical for maximal viability.
Changes in the Expression of Junctional and Nonjunctional Complex Component Genes When Inter‐Sertoli Tight Junctions Are Formed In Vitro
Authors: WONG CONNIE C. S. , CHUNG SANNY S. W. , GRIMA JOSEPHINE , ZHU LI‐JI , MRUK DOLORES , LEE WILL M. , CHENG C. YAN
Published: October 2023

ABSTRACT:Throughout spermatogenesis, germ cells move progressively from the basal to the adluminal compartment, which is accompanied by continual disassembly and reassembly of intercellular junctions suggesting germ cell movement is composed of intermittent phases of junction disassembly and reassembly. A study was performed to correlate the expression of junctional‐complex components (such as zonula occludens‐1 [ZO‐1], a tight‐junction component protein) and nonjunctional complex components (such as urokinase‐type plasminogen activator [uPA], a serine protease; cathepsin L, a cysteine protease; α2‐macroglobulin, a nonspecific protease inhibitor; and cystatin C, a cysteine protease inhibitor) at the time when inter‐Sertoli tight junctions were established in vitro. This is an attempt to investigate whether the expression of nonjunctional component genes also correlates with the formation of inter‐Sertoli tight junctions in vitro. This is part of an effort to understand the physiologic elements of germ cell movement in the epithelium. Sertoli cells cultured in vitro are known to undergo programmed cell death. To ensure that the changes in target gene expression were not the result of apoptosis, Sertoli cells were cultured in vitro at densities of 0.25, 0.75, and 3 × 106cells/cm2for up to 7 days on bicameral culture units coated with Matrigel (Collaborative Research) and were assessed by morphologic analysis and agarose gel electrophoresis. It was noted that many of the Sertoli cells cultured at 3×106cells/cm2underwent apoptosis by day 7, in contrast to cultures at 0.25 and 0.75 × 106cells/cm2illustrating the Sertoli cell number per unit of area may be an important parameter to be considered when studying Sertoli cell function in vitro. Also, it was shown that the expression of ZO‐1 increased significantly between days 2 and 3 prior to the establishment of inter‐Sertoli tight junctions assessed by transepithelial resistance measurement (TER), which illustrates that ZO‐1 can be used as a marker to monitor this cellular event. More interestingly, there was also a transient increase in the expression of uPA and cathepsin L between days 2 and 3 at the time preceding the formation of tight junctions. In Sertoli cells cultured at low density (2 × 104cells/cm2), when a confluent monolayer of cells could not form, there were no changes in the expression of either ZO‐1, uPA, or cathepsin L throughout the 7‐day culture period. These results show that the establishment of specialized junctions, such as tight junctions between Sertoli cells in vitro, may require the participation of both junctional and nonjunctional complex components.
Factors Involved in the Biochemical Etiology of Human Seminal Plasma Hyperviscosity
Authors: MENDELUK GABRIELA , FLECHA F. LUIS GONZÁLEZ , CASTELLO PABLO R. , BREGNI CARLOS
Published: October 2023

ABSTRACT: Semen rheology was studied to elucidate the biochemical basis of seminal plasma hyperviscosity. Semen proved to fit in with a power law model, by presenting a pseudoplastic behavior. Apparent viscosity at 230 s−1 and 25°C (ηa) was 4.3 ± 0.2 cp and 5.4 ± 0.4 cp in normal and high‐consistency semen, respectively. The effect of enzymes and mucolytic agents on human seminal plasma viscosity were evaluated by incubating normal and hyperviscous semen pool aliquots with trypsin, dithiothreitol, EDTA, α‐amylase and deoxyribonuclease I. After incubation, trypsin treatment reduced a by 36% in normal semen and by 44% in hyperviscous semen. There was a decrease in ηafollowing incubation of hyperviscous samples with dithiothreitol (33%) and α‐amylase (44%) that was not observed in the normal consistency samples. No decrease was observed in ηa after EDTA or DNAse treatment of both groups. Comparison of normal and hyperviscous seminal plasmas revealed no difference in the concentration of total proteins, DNA, or in the percentage of water content. These findings indicate that the primary substances responsible for basic normal semen rheologic behavior are proteins. A comparison of rheological properties between normal and hyperviscous semen samples indicates the existence of a highly organized network in the latter group, in which disulfide bonds and oligosaccharide chains complexed to the peptide core may play a key role.
Postvasectomy Alterations in Protein Synthesis and Secretion in the Rat Caput Epididymidis Are Not Repaired After Vasovasostomy
Authors: TURNER T. T. , RILEY T. A. , VAGNETTI M. , FLICKINGER C. J. , CALDWELL J. A. , HUNT‡ D. F.
Published: October 2023

ABSTRACT: Many men who have undergone vasectomy later request vasovasostomy. Unfortunately, significant numbers of these men remain infertile despite the reestablishment of patent ducts. This report examines the possibility that epididymal function remains compromised after vasovasostomy in the rat by examination of quantifiable, in vivo protein synthesis and secretion in the caput epididymidis. Rats were studied 30 days after vasectomy, 30 days after a vasovasostomy (which was performed 30 days after vasectomy), or after sham operations. Epididymal lumen fluids (LF) were collected by micropuncture after 3 hours' in vivo microperifusion of tubules with 35S‐amino acids. Proteins were separated by 2‐dimensional electrophoresis and were detected by Coomassie blue staining. Synthesized proteins in tubule extract and synthesized and secreted proteins in LF were detected by autoradiography and image analysis. Specific proteins that appeared to be affected by vasectomy‐vasovasostomy were identified by internal sequence analysis. LF contained an average of 87 detectable proteins synthesized and secreted in the control caput. Nineteen of the most prominent LF proteins were selected for more focused study. The most prominent proteins were clusterin, cysteine‐rich secretory protein (CRISP)‐1, and epididymal retinoic acid binding protein. Among these, CRISP‐1 remained reduced in LF after vasovasostomy. Two more minor proteins that remained reduced after vasovasostomy were identified as prostaglandin D2 synthase and phosphatidylethanolamine‐binding protein. All 3 of these proteins occur in the epididymides of multiple species and have been associated with sperm fertilizing capacity.
Assessment of the Acrosomal Status of Ram Spermatozoa by RCA Lectin‐Binding and Partition in an Aqueous Two‐Phase System
Authors: MARTÍ JOSE I. , CEBRIÁN‐PÉREZ JOSE A. , MUIÑO‐BLANCO TERESA
Published: October 2023

ABSTRACT: The acrosome reaction is an important marker for sperm function. Because different laboratory techniques may be used to detect this exocytotic process, the objective of this study was to investigate the use of fluoresceinated lectins to assess the acrosomal status of nonpermeabilized ram spermatozoa. In addition, we used centrifugal countercurrent distribution (CCCD) in an aqueous 2‐phase system to assess the sperm surface modifications associated with the acrosome reaction by observing changes in their partition behavior. We analyzed the binding of 5‐fluorescein isothiocyanate (FITC)‐conjugated lectins to ram sperm to select a lectin that bound preferentially to the acrosomal region, which would allow differentiation of acrosome‐intact from acrosome‐damaged ram spermatozoa. Ricinus communis agglutinin (RCA) bound intensely to the anterior and weakly to the equatorial acrosomal regions. Acrosomal labeling changed when spermatozoa were induced to Acrosome‐react with calcium ionophore A23187. RCA acrosomal labeling significantly increased (P < .0001) after incubation (84% versus 28% in control samples). To determine if RCA lectin labeling could be used to assess the acrosomal status of fresh ram spermatozoa in suspension, we compared the percentage of acrosome‐reacted sperm detected by the carboxyfluorescein diacetate/propidium iodide (CFDA/PI) double‐fluorescent staining with the percentage detected by FITC‐RCA labeling. The incidence of acrosome‐reacted spermatozoa detected by CFDA/PI was not significantly different (P = .704; 13 comparisons in 6 different experiments) from the incidence of spermatozoa detected by FITC‐RCA staining. The evaluation of the spontaneous acrosome reaction by RCA labeling (5.83%) was not significantly different (P = .644) from that assessed by CFDA/PI (6.88%). The percentage of induced acrosome reactions detected by CFDA/PI staining (56%) significantly correlated (P < .0001; r = 0.876) with that detected by RCA labeling (56.67%). We simultaneously carried out a comparative CCCD in an aqueous 2‐phase system to analyze sperm surface changes associated with the acrosome reaction. Results revealed that sperm surface hydrophobicity decreased in samples that had been incubated with ionophore compared with the untreated‐control samples. Likewise, RCA binding after CCCD showed that all acrosome‐reacted cells were stained, whereas only 42% of cells were lectin‐labeled in the untreated semen sample. This change in lectin reactivity of acrosome‐reacted spermatozoa signals the presence of some deep membrane or intracellular residues that would affect partitioning. Therefore, the FITC‐RCA—labeling procedure can be used to accurately assess the acrosomal status of ram spermatozoa in suspension.
Glycerol Disrupts Tight Junction—Associated Actin Microfilaments, Occludin, and Microtubules in Sertoli Cells
Authors: WIEBE JOHN P. , KOWALIK AGNES , GALLARDI ROBIN L. , EGELER OLIVER , CLUBB BRYAN H.
Published: October 2023

ABSTRACT: Intratesticular injections of glycerol have been shown to result in a marked and prolonged reduction of spermatogenesis, accompanied by increased permeability of the blood‐testis barrier. Because the permeability of the blood‐testis barrier is regulated by Sertoli cell tight junctions, and tight junction organization is regulated by the cytoskeleton, we undertook to examine the effects of glycerol treatment on cytoskeletal actin microfilaments and microtubules, and on the tight junction protein, occludin, in Sertoli cells. Adult rats received a single intratesticular injection of either saline (controls) or a 10% glycerol solution. At 24 hours and 7, 15, and 21 days after injection, testes were collected and prepared for routine histology, cryosectioning, or whole seminiferous tubule immunohistochemical staining; and the preparations were viewed by light and confocal microscopy. In saline‐injected testes, Sertoli cells had a cytoskeletal and junctional organization that resembled that of normal testes. F‐actin microfilaments, located in the basal region, were arranged in regular bundles or chords that circumscribed the perimeter of each Sertoli cell at the level of the tight junction. Occludin colocalized with tight junction—associated actin filament distribution and microtubules formed a geometric array associated with spermatogenic cells. In contrast, in glycerol‐treated Sertoli cells, microfilament and micro‐tubule organization and occludin distribution were partially or completely disrupted. From these results we conclude that glycerol treatment either directly or indirectly disrupts tight junction—associated F‐actin and occludin and tubulin organization in rat Sertoli cells. Perturbation of the tight junction—associated proteins could explain the increase in permeability of the blood‐testis barrier observed after glycerol treatment. Impaired spermatogenesis following glycerol treatment is likely a consequence of a leaky blood testis barrier and disrupted Sertoli cell cytoskeleton. Glycerol injections may serve as a useful tool in studying the relationship between cytoskeletal organization and the stabilization of Sertoli—Sertoli cell junctions.
High Catalase Content of Rabbit Semen Appears to be Inherited
Authors: FOOTE ROBERT H. , HARE ELIZABETH
Published: October 2023

ABSTRACT: Reactive oxygen species, as hydrogen peroxide, can be damaging to sperm. Most species have little protective catalase in their semen, but rabbit semen contains substantial amounts of catalase. The objective of the present study was to characterize a substantial number of Dutch rabbits for seminal content of catalase and determine whether differences were inherited. Usually, 4 or more semen samples were analyzed per male. Catalase was measured by a gasometric procedure. In Experiment 1, the correlation between duplicate determinations was r = .99, and between 2 sets of semen samples from 55 males it was r = .95. There was a significant difference (P < .05) among pairs of males from 6 litters. In Experiment 2, semen from each of 11 males collected at an interval of 1 year contained an average of 13 and 12 units of catalase per mL of semen in consecutive years. The correlation between pairs of samples was r = .85. This indicated that the condition was permanent, and possibly genetically controlled. Experiment 3 analyzed the catalase content of semen from 7 sires and 32 sons. The heritability of seminal catalase concentration was 0.48. These studies indicate that rabbit seminal plasma is high in catalase, and that a substantial portion of the differences among males are under genetic control.
Long‐Term Effects of Triptolide on Spermatogenesis, Epididymal Sperm Function, and Fertility in Male Rats
Authors: HUYNH PHUONG N. , HIKIM AMIYA P. SINHA , WANG CHRISTINA , STEFONOVIC KSENIJA , LUE YAN HE , LEUNG ANDREW , ATIENZA VINCE , BARAVARIAN SIMA , REUTRAKUL VICHAI , SWERDLOFF RONALD S.
Published: October 2023

ABSTRACT: Prior studies had suggested that triptolide, a diterpene triepoxide isolated from a Chinese medicinal plant, might be an attractive candidate as a post‐testicular male contraceptive agent. Despite the promise that triptolide would not affect testis function, nagging concerns remained that a delayed onset of testicular effect might exist. The objectives of this study were to assess the effects of relatively longer treatment duration of triptolide on fertility, spermatogenesis, and epididymal sperm pathophysiology; and to evaluate the reversibility of these effects after the cessation of treatment. Adult male Sprague‐Dawley rats were fed daily with either 30% gum acacia as a vehicle control (n = 12) or 100 μg/kg body weight (BW) of triptolide for 82 days (n = 12) followed by a recovery period of up to 14 weeks (n = 6). At the end of the treatment period, all rats treated with triptolide were sterile. Cauda epididymal sperm content decreased by 84.8% and sperm motility was reduced to zero. In addition, virtually all cauda epididymal sperm in the triptolide‐treated group exhibited severe structural abnormalities. The most striking changes observed were head‐tail separation, premature chromatin decondensation of sperm nuclei, a complete absence of the plasma membrane of the entire middle and principle pieces, disorganization of the mitochondrial sheath, and aggregation of many sperm tails. Longer treatment duration of triptolide also affected spermatogenesis, with marked variability in the response of individual animals. The degree of damage ranged from apparently normal‐looking seminiferous tubules to flattened seminiferous epithelium lined by a single layer of cells consisting of Sertoli cells and a few spermatogonia. Affected tubules exhibited intraepithelial vacuoles of varying sizes, multinucleated giant cells, germ cell exfoliation, and tubular atrophy. Recovery occurred as early as 6 weeks after cessation of treatment. By 14 weeks, 4 out of 6 triptolide‐treated males were fertile and the females that were impregnated by 3 out of 4 triptolide‐treated male rats produced apparently normal litters. These results suggest that triptolide has 2 phenotypic effects on mature and maturing germ cells. The first action appears earlier and manifests mainly in epididymal sperm. The second action presumably is directly on germ cells in testis and causes a variable impairment of spermatogenesis that may not be completely reversible. It is unclear if the earlier effect is a delayed manifestation of subtle testicular injury or post‐testicular action.
Immunolocalization of NBC3 and NHE3 in the Rat Epididymis: Colocalization of NBC3 and the Vacuolar H+‐ATPase
Authors: PUSHKIN ALEXANDER , CLARK IMRAN , KWON TAE‐HWAN , NIELSEN SØREN , KURTZ IRA
Published: October 2023

ABSTRACT: In the male reproductive tract, the epididymis plays an important role in mediating transepithelial bicarbonate transport and luminal acidification. In the proximal vas deferens, a significant component of luminal acidification is Na+‐independent, and mediated by specific cells that possess apical vacuolar proton pumps. In contrast, luminal acidification in the cauda epididymidis is an Na+‐dependent process. The specific apical Na+‐dependent H+/base transport processes) responsible for luminal acidification have not been identified. A potential clue as to the identity of these apical Na+‐dependent H+/base transporter(s) is provided by similarities between the transport properties of the epididymis and the mammalian nephron. Specifically, the H+/base transport properties of caput epididymidis resemble the mammalian renal proximal tubule, whereas the distal epididymis and vas deferens have characteristics in common with renal collecting duct intercalated cells. Given the known expression of the Na+/H+ antiporter, NHE3, in the proximal tubule, and of the electroneutral sodium bicarbonate cotransporter, NBC3, in renal intercalated cells, we determined the localization of NHE3 and NBC3 in various regions of rat epididymis. NBC3 was highly expressed on the apical membrane of apical (narrow) cells in caput epididymidis, and light (clear) cells in corpus and cauda epididymidis. The number of cells expressing apical NBC3 was highest in cauda epididymidis. The localization of NBC3 in the epididymis was identical to the vacuolar H+‐ATPase. The results indicate that colocalization of NBC3 and the vacuolar H+‐ATPase is not restricted to kidney intercalated cells. Moreover, the close association of the two transporters appears to be a more generalized phenomenon in cells that express high levels of vacuolar H+‐ATPase. Unlike NBC3, NHE3 was most highly expressed on the apical membrane of all epithelial cells in caput epididymidis, with less expression in the corpus, and no expression in the cauda. These results suggest that apical NBC3 and NHE3 potentially play an important role in mediating luminal H+/base transport in epididymis.
DNA Integrity in Human Spermatozoa: Relationships With Semen Quality
Authors: IRVINE D. STEWART , TWIGG JEREMY P. , GORDON EMMA L. , FULTON NORMA , MILNE PHILIP A. , AITKEN R. JOHN
Published: October 2023

Abstract: The literature contains conflicting evidence regarding the existence of DNA damage in spermatozoa from infertile male patients. To examine this phenomenon, we have studied ejaculated spermatozoa from normozoospermic semen donors and from a group of the unselected male partners of couples attending an infertility clinic for initial investigation. Classical semen analysis according to World Health Organization (WHO) guidelines was undertaken with computer‐assisted sperm analysis (CASA). Spermatozoa were prepared by sequential washing and centrifugation and were analyzed for DNA fragmentation using three assays: 1) a single‐cell gel electrophoresis (comet) assay, 2) in situ nick translation with prior chemical decondensation (ISNT‐decondensed), and 3) in situ nick translation without prior chemical decondensation (ISNT‐condensed). In addition, reactive oxygen species (ROS) generation by spermatozoa was measured, and seminal plasma was analyzed for its total reactive antioxidant potential (TRAP). When the donor and patient groups were compared, the latter had lower levels of semen quality and higher levels of DNA damage, which was particularly apparent using the comet assay. Highly significant negative correlations were observed between DNA fragmentation, detected by all three assays, and semen quality, particularly sperm concentration. In addition, multiple regression analysis indicated that other attributes of semen quality, such as sperm movement and ROS generation, were also related to DNA damage. We conclude that a significant proportion of infertile men have elevated levels of DNA damage in their ejaculated spermatozoa.
In Vitro and Xenogenous Capacitation‐Like Changes of Fresh, Cooled, and Cryopreserved Stallion Sperm as Assessed by a Chlortetracycline Stain
Authors: PARKER NIKOLA A. , BAILEY THOMAS L. , BOWEN JOHN M. , LEY WILLIAM B. , PURSWELL BEVERLY J. , DASCANIO JOHN J.
Published: October 2023

Abstract:Like the human female, the mare experiences reproductive tract pathology that may sometimes be circumvented by the use of assisted reproductive technologies (ARTs). One such technology, gamete intrafallopian transfer (GIFT), may be used in mares that exhibit ovulatory, oviductal, or uterine abnormalities that limit the use of common ARTs, such as embryo transfer. Homologous GIFT has been successfully performed in the horse; however, the logistics, costs, and associated risks of surgically transferring gametes to the oviducts of a recipient mare are considerably high. Use of a less costly species in a heterologous or xenogenous procedure would therefore be beneficial. This study represents the preliminary investigation into the use of sheep as recipients for xenogenous GIFT procedures using equine gametes. We investigated the capacitation response of fresh, cooled, or frozen stallion sperm after 1) in vivo incubation in the reproductive tract of estrous and anestrous ewes as well as 2) in vitro incubation in a modified Krebs/ Ringer extender at 37°C with and without the addition of heparin at 10 IU/mL for up to 8 hours. A chlortetracycline (CTC) fluorescent stain was used to assess the capacitation response of sperm. Findings indicated that oviductal fluid samples recovered from estrous ewes had significantly higher numbers of sperm exhibiting capacitation‐like staining patterns when compared to samples recovered from anestrous ewes (P < .05). Fresh semen yielded higher capacitation‐like staining patterns after in vivo incubation than did frozen‐thawed or cooled samples. A transition from majority CTC unreacted sperm to majority CTC non—acrosome intact sperm was demonstrated for both in vivo and in vitro studies. In vitro incubation of stallion sperm with heparin did not result in an increased capacitation‐like staining response over time when compared with nonheparinized samples. Results from this study suggest that xenogenous capacitation of stallion sperm may occur in the estrous ewe.
Comparison of Zinc Concentrations in Blood and Seminal Plasma and the Various Sperm Parameters Between Fertile and Infertile Men
Authors: CHIA SIN‐ENG , ONG CHOON‐NAM , CHUA LAY‐HA , HO LEE‐MEE , TAY SUN‐KUIE
Published: October 2023

Abstract: The aim of the study was to examine the relationships between concentrations of zinc in blood and seminal plasma and sperm quality among infertile and fertile men. One hundred seven male (infertile group) partners of couples who were undergoing investigation for infertility with no known cause for the infertility and 103 men (fertile group) whose wives were pregnant at the time of the study were recruited. The subjects' blood and seminal plasma concentration of zinc were determined by atomic absorption spectroscopy. Except for semen volume, all the other semen parameters for the infertile men were significantly lower than those for the fertile group. The geometric means of the seminal plasma zinc concentration were significantly lower in the infertile group compared with those in the fertile group; 183.6 mg/L (range, 63–499) versus 274.6 mg/L (range, 55–420). There were no significant differences in the geometric means of the blood zinc concentration between the 2 groups. Seminal plasma zinc concentration was significantly correlated with sperm density (r = 0.341, P < .0001), motility (r = 0.253, P < .0001), and viability (r = 0.286, P < .0001). On the basis of the findings of this study and those of other reports, zinc may contribute to fertility through its positive effect on spermatogenesis.
Spontaneous Hyperplasia of the Ventral Lobe of the Prostate in Aging Genetically Hypertensive Rats
Authors: GOLOMB ELIAHU , ROSENZWEIG NURIT , EILAM RAYA , ABRAMOVICI ARMAND
Published: October 2023

Abstract: Recent studies have shown that the prostatic autonomic innervation takes part in its homeostasis and growth. Other works showed that spontaneously hypertensive rats (SHR) show excessive sympathetic activity, accompanied by lower urinary tract symptoms, increased growth capacity of prostatic stromal cells, and increased levels of androgens and their receptors. Furthermore, young SHR were reported to present incipient stages of benign prostatic hyperplasia (BPH). The aim of the present study was to examine whether this strain indeed develops spontaneous BPH with age, and can thus serve as a genuine natural model for this disorder. For this purpose, ventral lobes of prostates of one‐year‐old, male SHR and their normotensive counterparts, Wistar Kyoto (WKY) rats, were examined histopathologically, and the degree of hyperplasia was evaluated according to a scorechart protocol (histoscore). SHR exhibited severe adenomatous spontaneous BPH, characterized by piling‐up of epithelial cells, with papillary formations, accompanied by a mild increase in the amount of fibrocytes and smooth muscle cells in the stroma. This was reflected by histoscore values of 38 ± 2. Thickening of prostatic arterioles also was noted, as well as mild chronic inflammatory exudate. WKY rats did not show any of these features of BPH despite their age (histoscore 17 ± 3, significantly different from that of SHR). We conclude that SHR can serve as a rodent model for the spontaneous development of BPH with age, most probably due to the excessive neuroendocrine activity characteristic of this rat strain.
Transdermal Electromotive Multi‐Drug Administration for Peyronie's Disease: Preliminary Results
Authors: MONTORSI FRANCESCO , SALONIA ANDREA , GUAZZONI GIORGIO , BARBIERI LUIGI , COLOMBO RENZO , BRAUSI MAURIZIO , SCATTONI VINCENZO , RIGATTI PATRIZIO , PIZZINI GIULIANO
Published: October 2023

Abstract: The purpose of this study was to clarify the actual therapeutic potential of a new transdermal drug delivery system (electromotive drug administration; EMDA) for selected patients with Peyronie's disease. Forty patients with Peyronie's disease were treated by electromotive administration of the 3‐drug association orgotein‐dexamethasone‐lidocaine in a double‐blind, placebo‐controlled, partial crossover study (study 1). Another 25 patients were treated by EMDA with a combination of verapamil‐dexamethasone in an uncontrolled study (study 2). Treatment sessions lasted 20 minutes each and took place 3 times a week for 3 weeks with a current of 3 mA. Patients were assessed before treatment and at 1‐and 3‐month follow‐up examinations. Assessments were based on sexual history, physical examination, and dynamic color Doppler ultrasonographic results. Adverse effects of EMDA were not reported. In study 1, the clinical results observed after treatment proved to be significantly better than those of the placebo. Penile pain disappeared in all patients in both studies. Penile lesion (nodule or plaque) either disappeared or significantly improved in 79% and 90% of patients treated by the 3‐and 2‐drug association, respectively. The improvement of penile deformity also was notable although it did not match the effect observed on penile nodules or plaque (62% and 88%, in studies 1 and 2, respectively). In both studies, more than 80% of patients reported a definite amelioration of penile rigidity, which paralleled the improvement of penile dynamic color Doppler ultrasonographic parameters. Overall, the combination of verapamil‐dexamethasone achieved better clinical results than the 3‐drug combination. Electromotive drug administration is a novel technique capable of safely achieving satisfactory results in selected patients with Peyronie's disease not only in terms of improvement of patient's symptoms but also due to the reduced need for penile surgery.
Androgen Receptor Gene Polymorphism and Prostate Zonal Volumes in Australian and Chinese Men
Authors: JIN B. , BEILIN J. , ZAJAC J. , HANDELSMAN D. J.
Published: October 2023

Abstract: Prostate diseases are age and androgen dependent. The evolution of clinically overt pathology requires decades of exposure to adult male levels of circulating testosterone, but the precise relationship between age and androgen circulation remains poorly understood. A marker of integrated androgen action over prolonged periods would therefore be a valuable tool for clinical and epidemiologic research into the origins of prostate disease. In order to evaluate these 2 factors, we have studied the CAG‐repeat length polymorphism of the androgen receptor gene and the size of the total, central, and peripheral zones of the prostate, estimated by planimetric ultrasound in 2 populations with widely different susceptibility to death from invasive prostate cancer. From a larger epidemiologic study of the effects of ethnicity and migration on the origins of prostate disease, a nestedcase control study was undertaken with 50 Chinese men living in Yue Yang, China and 50 non‐Chinese men living in Sydney, Australia. All men had undergone planimetric transrectal prostate ultrasound together with blood sampling to determine CAG‐repeat length by polymerase chain reaction and immunoassay of plasma testosterone, estradiol, dihydrotestosterone (DHT), sex hormonebinding globulin (SHBG), and prostate‐specific antigen (PSA). Australian men had larger central (7.9 ± 0.4 vs 3.3 ± 0.3 mL) and total (29.8 ± 1.2 vs 25.5 ±1.1 mL) but not peripheral (22.0 ± 0.9 vs 22.2 ± 0.8 mL) prostate volumes compared with Chinese men. Even after adjustment for differences in body size (the Australian men were taller and heavier), the central‐zone volume remained lower by ∼50% in Chinese men (P < 0.001), whereas testis and total‐prostate volumes were no longer significantly different. The length of CAG repeats was no different between Australian men (22.5 ± 0.5 repeats) and Chinese men (22.5 ± 0.5 repeats), and there was no correlation within or between populations in CAG repeats or any measure of prostate volume or hormones. DHT concentration was 20% lower in Chinese men compared with Australian men (1.6 ± 0.1 vs 2.0 ± 0.1 nmol/L, P = 0.005), a difference that persisted after age adjustment (P = 0.039) but that was removed by adjustment for differences in total‐prostate size (P = 0.12). Blood testosterone, estradiol, SHBG, and PSA concentrations were not different between the 2 populations. Hence, the hypothesis is refuted that the CAG repeat polymorphism in the androgen receptor gene (within the nonpathologic range) and the central‐prostate zone volume might be markers of long‐term androgen sensitivity. Whether either factor alone may constitute a marker of androgen sensitivity remains to be established by other means, and a long‐term marker of integrated androgen action suitable for clinical and epidemiologic research is still lacking.
Morphologic Changes in Efferent Ductules and Epididymis in Estrogen Receptor‐α Knockout Mice
Authors: HESS REX A. , BUNICK DAVID , LUBAHN DENNIS B. , ZHOU QING , BOUMA JERRY
Published: October 2023

Abstract: Estrogen has been shown to have an important role in fluid reabsorption in efferent ductules of the testis. Our previous study of the estrogen receptor‐α knockout mouse (ERKO) showed that the efferent ductules and rete testis were primary targets of estrogen receptor function. In the present study, a more comprehensive evaluation of the ERKO male reproductive tract was performed to determine the severity of effects in efferent ductules as well as the epididymis. The following observations were found in ERKO males: 1) blind‐ending efferent ductules were more prevalent in ERKO than in wild type (WT) tissues; 2) glycogen‐containing cells were observed at the rete testis—efferent ductule junction; 3) the tubular diameters of the efferent ductules and initial segment epididymides were dilated; 4) efferent ductules were dilated between 130 to 300% over wild type ductules; 5) efferent ductule epithelial height was reduced nearly 50%; 6) microvilli of nonciliated cells of efferent ductules were 64% shorter in length; 7) cilia were reduced in number; 8) initial segment epithelium was displaced into regions adjacent to the rete testis and in short segments of the common region of efferent ductule; 9) apical, narrow, and clear cells of the epididymis also were abnormal in some regions; 10) in the corpus and cauda regions, sperm granulomas were noted in one third of the ERKO males. In conclusion, the entire reproductive tract is affected in ERKO males. The cells showing the greatest effects were estrogenreceptor—positive cells. It appears that in the ERKO mouse there are developmental anomalies that must be considered separately from adult dysfunctional changes in the male reproductive tract.
Semen Quality and Human Fertility: A Prospective Study With Healthy Couples
Authors: ZINAMAN MICHAEL J. , BROWN CHARLES C. , SELEVAN SHERRY G , CLEGG ERIC D
Published: October 2023

Abstract: Measures of semen quality are used as surrogate measures of male fertility in clinical andrology, reproductive toxicology, epidemiology, and risk assessment. However, only limited data are available to relate those measures to fertility. This prospective study with 210 reproductive‐age couples was conducted to provide information on the value of semen quality measures for predicting human male fertility potential and for development of models to estimate the effects of changes in semen quality on fertility in a given population for risk assessment. Couples without known risk factors for infertility and who had discontinued contraception to have a child were accepted. The study followed each couple for up to 12 menstrual cycles while they attempted to conceive and evaluated semen quality measures from multiple ejaculates per man with known abstinence intervals. For each cycle, the day of ovulation was predicted, and the couple was advised to have intercourse multiple times on that day and on the days around it. Among the demographic variables assessed, parity, contraception status prior to entering the study, male education level, and male smoking were associated significantly with 12‐cycle pregnancy rate. Several semen quality measures also were associated significantly with pregnancy rate, with percentage morphologically normal sperm by strict criteria and measures involving total number of sperm showing particularly strong associations. Localized regression‐smoothing plots of semen quality data against proportion of couples pregnant suggested levels below which fertility declines for several semen quality measures. These results have applications in both clinical andrology and in assessment of risk to male fecundity from environmental or pharmaceutical exposures. In particular, they contribute information on behavior of fertility with varying semen quality and can allow development of models to predict effects on fertility in populations from decrements in semen quality.
Bombesin Modulates the Association of Src with a Nuclear 110‐kd Protein Expressed in Dividing Prostate Cells
Authors: ALLARD PIERRE , BEAULIEU PATRICK , APRIKIAN ARMEN , CHEVALIER SIMONE
Published: October 2023

ABSTRACT: Polyclonal antibodies produced against a peptide derived from the Fer tyrosine kinase sequence also specifically recognized a 110‐kd protein (p110) up‐regulated in dividing versus resting dog prostate epithelial cells in vitro. In vivo in the dog prostate, p110 expression was detected when basal cell metaplasia was induced by estrogens after castration but not when renewing the differentiated epithelium with androgens. It was also detected in extracts from the human prostatic carcinoma cell lines LNCaP, DU145, and PC‐3, and from 6 out of 11 human prostate cancer tissues analyzed, but not from normal or hyperplastic glands. The tyrosine kinase Src was shown by coimmunoprecipitation to associate with p110, and this interaction was positively modulated by bombesin stimulation of PC‐3 cells. However, p110 was not tyrosine phosphorylated. Moreover, it was mainly distributed in the nuclear fraction. This nuclear p110 protein, expressed in dividing prostate epithelial cells and in human prostate cancer cells and tissues, could thus be a downstream mediator of bombesin‐signaling pathways, acting via its association with Src.
Immunolocalization of CA II and H+V‐ATPase in Epithelial Cells of the Mouse and Rat Epididymis
Authors: HERMO LOUIS , ADAMALI HUZAIFA I. , ANDONIAN SERO
Published: October 2023

ABSTRACT:Acidification of the epididymal lumen has been suggested to play an important role in sperm functions; however, the cell types, pumps, and mechanisms involved have not been fully addressed. In this study, carbonic anhydrase II (CA II) and a 67‐kd subunit ofNeurospora crassavacuolar proton adenosinetriphosphatase (H+V‐ATPase) pump were immunolocalized using light microscopy and electron microscopy (EM) in the epididymis of rats and mice. In both animals, narrow cells, identified in the initial segment and intermediate zone of the epididymis, contained numerous small vesicles in their apical region, often cup‐shaped in appearance. In the mouse but not rat, these cells also possessed numerous cisternae of smooth endoplasmic reticulum, suggesting steroid synthesis; and cytoplasmic blebs of their apical cell surface, which appeared to detach, suggesting apocrine secretion. Anti‐CA II antibody was immunocytochemically localized in the light microscope within narrow cells but not over any other cell types of the entire epididymis. Anti‐H+V‐ATPase antibody was also localized in narrow cells of the initial segment and intermediate zone; as well as clear cells of the caput, corpus, and cauda regions. Using EM, gold particles for anti‐CA II and H+V‐ATPase antibodies were noted in the apical region of narrow cells in relation to the numerous, small, cup‐shaped vesicles. Although CA II was mainly located in the cytosol near these vesicles, H+V‐ATPase appeared on their delimiting membrane and on the apical plasma membrane of these cells. A similar distribution was noted for H+V‐ATPase in clear cells. The nature of the small vesicles of the apical region of narrow cells was examined with electron‐dense fluid phase tracers that were introduced into the epididymal lumen. The tracers appeared within these vesicles and a few endosomes 1 hour after injection, suggesting that they contact the apical plasma membrane. Since these vesicles are also related to CA II and H+V‐ATPase, the data suggests that, as the site of proton production, the vesicles recycle to and from the apical cell surface, and in this way, deliver protons to the epididymal lumen for acidification. Clear cells and their expression of H+V‐ATPase may also serve in this function. In summary, both narrow and clear cells appear to be involved in luminal acidification, an activity that may be essential for sperm as they traverse and are stored in the epididymis.
Cryopreservation of a Small Number of Fresh Human Testicular Spermatozoa and Testicular Spermatozoa Cultured In Vitro for 3 Days in an Empty Zona Pellucida
Authors: LIU JIAEN , ZHENG XUE‐ZHONG , BARAMKI THEODORE A. , COMPTON GAIL , YAZIGI RICARDO A. , KATZ EUGENE
Published: October 2023

ABSTRACT: It is known that the motility of human testicular sperm can be improved when they are cultured in vitro for a few days. The purpose of this study was to determine whether it is better to freeze human testicular spermatozoa on the day of biopsy (fresh) or after they were cultured for 3 days. A modified, single‐sperm freezing technique was used in this study. The study consisted of two parts: (1) ejaculated spermatozoa were used to examine the influence of different concentrations of glycerol and synthetic serum substitute (SSS) on the survival rate after cryopreservation, and (2) the survival rates between cryopreserved fresh testicular spermatozoa (Group 1) and testicular spermatozoa that were cultured for 3 days before freezing (Group 2) were compared. Empty zonae pellucidae were obtained from mouse eggs. Five to 10 motile spermatozoa were selected and injected into an empty zona pellucida. For freezing, the zona pellucida with spermatozoa was transferred into a HEPES‐buffered human tubal fluid containing different concentrations of glycerol and kept at room temperature for 10 to 15 minutes, and then loaded into a 0.25‐ml‐plastic straw. The straws were exposed to liquid nitrogen vapor for 2 hours and then plunged into liquid nitrogen. For thawing, the straws were taken out of liquid nitrogen and placed into a 37°C waterbath for 25 to 30 seconds. There was no statistically significant difference in survival rates between 3% and 10% SSS with different glycerol concentrations. There was no statistically significant difference in the survival rates of spermatozoa between Group 1 and Group 2 after cryopreservation. It appears that in vitro culture of testicular spermatozoa before freezing does not increase survival rate.