bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Alprazolam on Respiratory Drive, Anxiety, and Dyspnea in Chronic Airflow Obstruction: A Case Study
Authors: Greene Jerry G. , Pucino Frank , Carlson James D. , Storsved Mark , Strammen Gordon L.
Published: December 2015

Alprazolam, an anxiolytic benzodiazepine, has a pharmacologic profile similar to that of diazepam. An intermediate half‐life of 10–12 hours and a comparatively brief duration of activity relative to other anxiolytic benzodiazepines justified evaluation of a 0.5–mg test dose in an anxious patient with chronic obstructive lung disease. Subjective indexes, breath‐by‐breath respiratory drive response to hypercapnia, and blood alprazolam concentrations were determined before and after dosing. Subjective testing included a visual analog dyspnea scale, the state anxiety inventory, and subjective feelings visual analog scales (represented by alertness, calmness, and level of contentment). After dosing, the patient was better able to tolerate the rebreathing study technique. Statistically significant improvements in dyspnea (t — 10.20; p0.0005), anxiety (t — 45.85; p < 0.0001), alertness (t — 13.04; p < 0.0001), cententedness (t — 12.27; p < 0.0001), and calmness (t — 8.05; p < 0.0001) occurred after alprazolam administration. Drive to breathe, as determined by mouth occlusion pressure and minute ventilation, was not statistically different before and after dosing. No adverse effects were reported or observed. Further study is warranted.
Concentrations of Immunoreactive Inhibin in Seminal Plasma and Serum from Normospermic, Oligospermic, Vasectomized, Klinefelter's Syndrome, and Sertoli‐cell–only Syndrome Subjects
Authors: ASCH RICARDO H. , VAZE A. Y. , THAKUR A. N. , SHETH ANIL R.
Published: December 2015

Semen samples and blood were collected from 57 men: 23 normospermic, 20 oligospermic, 10 vasectomized, two with Klinefelter's syndrome, and two with Sertoli‐cell–only syndrome. Inhibin was measured by radioimmunoassay techniques as reported previously. Immunoreactive inhibin concentrations in seminal plasma were found to be 8000 to 10, 000 times higher than in blood. A significant correlation (P <0.005) was observed between the serum and seminal plasma concentrations in all groups. No direct correlation was observed between immunoreactive inhibin concentrations in serum or semen with number of sperm in the ejaculate or with sperm motility. Seminal plasma and serum immunoreactive inhibin levels in normospermic subjects were significantly higher than in oligospermic, vasectomized, and Klinefelter's syndrome subjects (P <0.001 and <0.01, respectively).
Rat Testicular Phospholipase A2 Activity: pH Optima, Its Cellular and Subcellular Distribution in the Gonad, and Some Factors That May Modulate Its Activity
Authors: ELLIS LEGRANDE C. , BOCCIO JAMES R. , CUNNINGHAM MICHAEL J. , GROESBECK MICHAEL D. , COSENTINO M. JAMES
Published: December 2015

A sensitive radiometric assay was developed for detecting phospholipase A1 and A2 (PLA1 and A2) activity. Four to five times more PLA2 activity was observed in rat testes than PLA1 activity. Two PLA2 were observed in the testis, one with an acid (3.5) and one with an alkaline (7.5–8.0) pH optimum. The acid pH optimal PLA2 was located in the interstitial and lysosomal fractions. The alkaline pH optimal PLA2 was localized in the germinal elements of the seminiferous tubules and in the lysosomalenriched and membrane‐enriched fractions. Triton X‐100 inhibited PLA2 at 10−2 M and inhibited PLA1 at 10−3. At 10−2 PA, triton X‐100 activated PLA1. EGTA inhibited PLA2 activity, whereas Ca++ at 10−2 to 10−3 M restored this activity. Corticosterone had no effect on PLA2 activity, but progesterone, dihydrotestosterone, and testosterone all stimulated the enzyme at lower concentrations (10−9‐10−7 M), with testosterone giving maximUm stimulation at a lower dosage (ie, 10−9 M compared to 10−7 M for dihydrotestosterone). Both androgens inhibited PLA2 activity at higher concentrations.
Evidence for a Role of Post‐Ovulatory Cumulus Components in Supporting Fertilizing Ability of Hamster Spermatozoa
Authors: BAVISTER BARRY D.
Published: December 2015

A completely chemically defined culture medium was used to investigate the role of egg‐cumulus complex (ECC) components in supporting sperm fertilizing ability. Defined sperm motility‐stimulating factors (hypotaurine and epinephrine), with polyvinylalcohol as the macromolecular component, were included in the defined medium. Freshly‐ovulated hamster ECCs were incubated with washed epididymal spermatozoa under different conditions designed to evaluate the ability of ECC components to support sperm capacitation, acrosome reactions, and the ability to penetrate the ova. The major conclusions from the data are that ECC components are capable of supporting these physiologic events, and that these components are present in the soluble (fluid) compartment of the ECC. This work is the first in a series of steps aimed at the localization, characterization, and eventual identification of the natural acrosome reaction‐inducing stimulus associated with the ECC.
Sperm‐Egg Interaction
Authors: ORGEBIN‐CRIST M.‐C. , FOURNIER‐DELPECH SUZANNE
Published: December 2015

A study in vitro of interactions between uncapacitated rat spermatozoa and freshly ovulated rat oocytes showed that testicular spermatozoa were unable to adhere to the zona pellucida. On the other hand, spermatozoa from the cauda epididymidis adhered readily to the zona surface. Experiments with cauda spermatozoa, immobilized by a preincubation at 0 C, indicated that they were still able to attach to the zona pellucida, suggesting that the inability of testicular spermatozoa to interact with the zona was not primarily due to their lack of motility but rather to a defect in adhesive properties of the plasma membrane. Preincubation of testicular spermatozoa, with a purified glycoprotein secreted by the epididymis (PES), increased somewhat the ability of testicular spermatozoa to interact with the zona surface. These results show that epididymal spermatozoa acquire the ability to recognize and interact with the zona pellucida as they pass through the epididymis, and suggest that sperm coating with an androgen‐dependent and specific glycoprotein, secreted by the epididymis, may regulate the initial contact with the egg investments.
Estrogen Synthesis by Immature Rat Sertoli CellsIn Vitro
Authors: SUÆREZ‐QUIAN CARLOS A. , DYM MARTIN , MAKRIS ANASTASIA , BRUMBAUGH JOANNE , RYAN KENNETH J. , CANICK JACOB A.
Published: December 2015

Past research has implicated both the Leydig and Sertoli cells as sources of testicular estrogens. To study this issue further, we investigated the ability of Sertoli cells isolated from the testes of immature (10‐day‐old) rats to synthesize estrogens in response to FSH in vitro. The aromatase activity of Sertoli cells was examined by comparing simultaneously two different methods: 1. the conversion of 19‐(6,7‐3H)‐hydroxyandrostenedione to (3H)estrone and (3H)estradiol, measured by silica gel thin layer chromatography; and 2. the conversion of unlabeled 19‐hydroxyandrostenedione to total estrogen (El + E2), measured by RIA.

Sertoli cells, cultured in the presence of substrate, exhibited aromatase activity in response to FSH. Values obtained for total estrogen synthesis, measured by direct tracer methodology, at FSH dosages of 0.5 μg/ml and 1.0 μg/ml, during a 24‐hour period of incubation, were 620 pg/mg and 580 pg/mg, respectively; and after a 48‐hour incubation, values obtained were 570 pg/ml and 560 pg/mg, respectively. When measured by RIA, similar results were obtained for estrogen synthesis at identical FSH dosages and times of incubation. A dose response of FSH‐stimulated aromatization was also obtained by RIA. Neither control nor LH‐treated cells, however, in the presence of substrate, produced detectable levels of labeled or immunoassayable estrogens.

Aromatase activity was also studied in the presence of a cold estrogen trap and measured by direct tracer methodology. In this case, estrogen synthesis was more than doubled. These data may suggest the further metabolism of estrogen by Sertoli cells. Finally, our results are discussed with respect to the role of the immature Sertoli cell in the synthesis of testicular estrogens.
Correlation Between Human Sperm Swelling in Hypoosmotic Medium (Hypoosmotic Swelling Test) and In Vitro Fertilization
Authors: VAN DER VEN HANS H. , JEYENDRAN RAJASINGAM S. , AL‐HASANI SAFAA , PEREZ‐PELAEZ MARIANO , DIEDRICH KLAUS , ZANEVELD LOURENS J. D.
Published: December 2015

Human ejaculates (n = 83) were analyzed for standard sperm parameters (concentration, motility, and morphology), as well as for the ability of the spermatozoa to react (swell) in a hypoosmotic medium (Jeyendran et al, 1984). Subsequently, the fertilizing capacity of the spermatozoa was tested by their ability to fertilize human oocytes in vitro. Although the sperm concentration was adjusted for in vitro fertilization, no adjustments were made for sperm motility and morphology. Correlation of the in vitro fertilizing capacity of the spermatozoa with the hypoosmotic swelling test (r = 0.56) was much higher than with standard sperm parameters (r varied from −0.04 to 0.25). Complete overlap was noted with standard semen parameters whether the ejaculate did or did not fertilize oocytes and ranged from very low to very high values in both cases. By contrast, all the semen samples that fertilized oocytes showed a 60% or higher reaction in the hypoosmotic swelling test, whereas the majority of the “infertile” semen samples showed less than 60% swelling. It therefore appears that, under the conditions of our studies, the hypoosmotic swelling test is a more accurate predictor of successful in vitro fertilization outcome than the conventional semen parameters. A combination of all parameters, however, is likely to be most useful. The hypoosmotic swelling test is simple and economical, and it is recommended that this test be further scrutinized for its value as an additional tool in the assessment of the in vivo fertilizing capacity of ejaculated spermatozoa.
Binding and Internalization In Vivo of [125I]hCG in Leydig Cells of the Rat
Authors: HERMO L. , LALLI M.
Published: December 2015

The present study was performed to demonstrate the binding, mode of uptake, pathway and fate of iodinated human chorionic gonadotropin ([125I]hCG) by Leydig cells in vivo using electron microscope radioautography. Following a single injection of [125I]hCG into the interstitial space of the testis, the animals were fixed by perfusion with glutaraldehyde at 20 minutes, 1, 3, 6 and 24 hours. The electron microscope radioautographs demonstrated a prominent and qualitatively similar binding of the labeled hCG on the microvillar processes of the Leydig cells at 20 minutes, 1, 3, and 6 hours. The specificity of the [125I]hCG binding was determined by injecting a 100‐fold excess of unlabeled hormone concurrently with the labeled hormone. Under these conditions, the surface, including the microvillar processes of Leydig cells, was virtually unlabeled, indicating that the binding was specific and receptor‐mediated. In animals injected with labeled hCG and sacrificed 20 minutes later, silver grains were also seen overlying the limiting membrane of large, uncoated surface invaginations and large subsurface vacuoles with an electron‐lucent content referred to as endosomes. A radioautographic reaction was also seen within multivesicular bodies with a pale stained matrix. At 1 hour, silver grains appeared over dense multivesicular bodies and occasionally over secondary lysosomes, in addition to the structures mentioned above, while at 3 and 6 hours, an increasing number of secondary lysosomes became labeled. At 24 hours, binding of [125I]hCG to the microvillar processes of Leydig cells persisted but was diminished, although a few endosomes, multivesicular bodies and secondary lysosomes still showed a radioautographic reaction. No membranous tubules that were seen in close proximity to, or in continuity with, endosomes and multivesicular bodies were observed to be labeled at any time interval. Likewise, an attempt to correlate silver grains with small coated or uncoated pits, the stacks of saccules of the Golgi apparatus and other Golgi‐related elements including GERL, proved unsuccessful, since these structures were mostly unlabeled. These in vivo experiments thus demonstrate the specific binding of [125I]hCG to the plasma membrane of Leydig cells predominantly on their microvillar processes, and the subsequent internalization of the labeled hCG to secondary lysosomes. In addition, binding and internalization of hCG persisted for long periods of time.
Computer‐Assisted Human Semen Analysis Sampling Errors and Reproducibility
Authors: GINSBURG KENNETH A. , MOGHISSI KAMRAN S. , ABEL ERNEST L.
Published: December 2015

Videomicrographic computer‐automated semen analysis systems allow quantitative description of sperm motility, velocity, progression, and head movement amplitude and frequency with unprecedented ease. The minimum number of spermatozoa needed for stable results, the variability of measurements and optimum methods of sampling the ejaculate were determined for one such system (Cell‐Soft, CRYO Resources, New York, NY). Sampling a minimum of 225 spermatozoa yields stable measurements, and analyzing four microscope fields in triplicate provides data with the lowest coefficient of variation. The variability attributable to the instrument itself was acceptable for all measurements (6.2% to 15.4%) except mean amplitude of lateral head displacement. Limitations of these results and the potential utility of videomicrographic sperm movement analysis are discussed.
Luteinizing Hormone Secretion by Male Rat Pituitary Cells Perifused In Vitro: Effect of Experimental Left Varicocele and Orchiectomy
Authors: PRYOR JON L. , KRIEG RICHARD J. , EVANS WILLIAM S. , BATSON JUDY , TURNER TERRY T.
Published: December 2015

It has been shown previously that experimental left varicocele in the rat, results in a bilateral decrease in intratesticular testosterone. In the present work, pituitary responsiveness to GnRH as a possible mediator of this effect has been examined. Unilateral varicoceles were created in adult rats. A second group of animals underwent a sham operation and a third underwent bilateral orchiectomy. Thirty days after surgery, rats from all three groups were sacrificed and their pituitaries were removed. Dispersed pituitary cells were perifused in Bio‐Gel columns with varying concentrations of GnRH. The concentration of LH in the collected eluent was determined by radioimmunoassay. The mean, overall GnRH‐stimulated LH immunoreactive secretion rate (ng/min/107 cells) by pituitary cells from rats with varicocele (0.062 ± 0.11) was no different from the overall release from the sham‐operated controls (0.051 ± 0.007). The dose‐response curves for GnRH‐stimulated release of LH by dispersed pituitary cells in the two groups also were not different. The overall GnRH‐stimulated LH release by cells from the orchiectomized rats (0.171 ± 0.032) was significantly greater than release by cells from the sham‐operated and varicocele rats, and the concentration‐response curve from the orchiectomy group was significantly elevated over those of the other two groups. These results indicate that GnRH‐stimulated immunoreactive LH release is not altered in rats with experimental left varicocele and, thus, is not the source of an endocrinopathy that leads to decreased intratesticular testosterone concentrations in these animals.
Development of Transmembrane Signaling in the Fetal Rat Leydig Cell
Authors: WARREN DWIGHT W.
Published: December 2015

Gonadotropin binding to the adult Leydig cell activates a GTP binding protein that interacts with adenylate cyclase to increase cAMP production within the cell. The increased production of cAMP stimulates steroidogenesis and leads to an increase in testosterone production and secretion. The fetal Leydig cell responds to LH with an increase in cAMP and testosterone production as early as 15.5 days of gestation, although the specific mechanism of transmembrane signaling has not been characterized. Fetal rat testis cells from 13.5–20.5 days of gestation were treated with dibutyryl cAMP (dbcAMP), cholera toxin, and hCG to determine the onset of steroidogenesis stimulation by activation of each moiety in the transmembrane signaling system of the fetal Leydig cell. Maximal stimulation at each age from 14.5 through 20.5 days of gestation was achieved with 1 mM dbcAMP, 500 ng/ml cholera toxin, or 10 ng/ml hCG. At 13.5 days of gestation, fetal testes did not produce any testosterone. These findings indicate that a cholera toxin‐sensitive, stimulatory guanine‐nucleotide regulatory protein is functional in the fetal Leydig cell as early as 14.5 days of gestation. The LH receptor becomes functional in the transmembrane signaling system of the fetal Leydig cell at 14.5 days of gestation.
Inhibins in the Male Göttingen Miniature Pig: Leydig Cells Are the Predominant Source of Inhibin B
Authors: JIN WANZHU , ARAI KOJI Y. , HERATH CHANDANA B. , KONDO MASAHIRO , ISHI HAZIME , TANIOKA YOSHIKUNI , WATANABE GEN , GROOME NIGEL P. , TAYA KAZUYOSHI
Published: December 2015

ABSTRACT: The expression of inhibin subunits in the testes of the Göttingen miniature pig was examined by in situ hybridization and immunohistochemistry. In addition, the major forms were determined by enzyme‐linked immunosorbent assay (ELISA). Strong positive immunostaining for the inhibin α subunit was observed in Sertoli and late‐stage germ cells, but it was weak in Leydig cells. However, Leydig cells showed strong positive staining for the βA subunit, but Sertoli cells and spermatogonia showed a weak reaction. Strong positive immunostaining for the βB subunit was observed in Leydig cells but spermatogonia showed weak staining for it. In contrast to the staining specificity of inhibin α and βA subunits, the βB subunit did not exhibit positive staining in Sertoli cells. In situ hybridization revealed that although the α subunit mRNA signal was highly expressed in all cell types, the reaction appeared to be stronger in Sertoli cells and spermatogonia than in Leydig cells. βA subunit mRNA expression was somewhat identical to that of the α subunit, however, germ cells showed a weak stain for it. A strong, positive mRNA signal for the βB subunit was confined to Leydig cells and late‐stage germ cells. ELISA results showed that concentrations of inhibin B and inhibin pro‐αC were high in the circulation and testes. In contrast, inhibin A levels in both plasma and testes were undetectable. The present results strongly suggest that inhibin B is the major form of circulating inhibin and that Leydig cells are the predominant source of this dimeric inhibin in male Göttingen miniature pigs. Furthermore, the germ cells also appear to be an important source of circulating inhibins.
Prenatal Exposure to Dexamethasone Alters Leydig Cell Steroidogenic Capacity in Immature and Adult Rats
Authors: PAGE KATHLEEN C. , SOTTAS CHANTAL M. , HARDY MATTHEW P.
Published: December 2015

ABSTRACT: This study examines the effects of prenatal exposure to dexamethasone (DEX) on postnatal testosterone production in male rats. Pregnant female rats were treated on gestation days 14–19 with DEX (100 μg/kg body weight per day; n = 9) or vehicle (n = 9). Results show that 35‐day‐old male offspring from DEX‐treated pregnant females (n = 42) had decreased levels of serum testosterone (45.6% lower, P< .05) compared with control offspring (n = 43), although serum luteinizing hormone (LH) levels were not significantly altered. These findings suggest that a direct programming of developing gonadal cells occurs in response to high levels of maternal glucocorticoid. Indeed, testosterone production was significantly reduced in Leydig cells isolated from immature offspring of DEX‐treated pregnant females compared with controls (48.3%, P < .001), and LH stimulation of these cells did not compensate for the lowered steroidogenic capacity. The hypothalamic‐pituitary‐adrenal axis was also affected, because significant reductions in both serum adrenocorticotropic hormone (ACTH; 26.2%, P < .001) and corticosterone (CORT; 32.3%, P < .001) were measured in DEX‐exposed immature male offspring. In contrast, adult male offspring from DEX‐treated dams had significantly higher levels of serum ACTH (39.2%, P <.001) and CORT (37.8%, P < .001). These same animals had higher serum testosterone (31.6%, P < .05) and a significant reduction in serum LH (30.8%, P < .001). Moreover, Leydig cells isolated from these adult offspring exhibited an increased capacity for testosterone biosynthesis under basal (38.6%, P < .001) and LH‐stimulated conditions (33.5%, P < .001). In summary, sustained changes in steroidogenic capacity were observed in male rats exposed to high levels of glucocorticoid during prenatal development. More specifically, DEX exposure in utero perturbed Leydig cell testosterone production in both pubertal and adult rats.
Requirement of Notch 1 and Its Ligand Jagged 2 Expressions for Spermatogenesis in Rat and Human Testes
Authors: HAYASHI TETSUO , KAGEYAMA YUKIO , ISHIZAKA KAZUHIRO , XIA GUANGBIN , KIHARA KAZUNORI , OSHIMA HIROYUKI
Published: December 2015

ABSTRACT: It has already been demonstrated that the Notch signaling system is essential for gametogenesis in the adult germ line of Caenorhabditis elegans. However, the role of the Notch signaling system in mammalian spermatogenesis has not been well investigated. Recently, it has been revealed that this signaling system is expressed in the mammalian testis by showing coexpression of Jagged 2 and its receptor, Notch 1, is consistent with Notch 1 being a cognate receptor for Jagged 2 in the mammalian testis. Therefore, we investigated expressions of messenger RNAs of Notch 1 and Jagged 2 in the testicular tissues of developing Sprague‐Dawley rats by reverse transcription‐polymerase chain reaction and Northern blot analysis, expressions of their proteins in the testicular tissues of developing rats, fertile human controls and infertile human patients with maturation arrest by immunohistochemistry, and effects of antibodies to this system by culturing rat testicular tissues with these antibodies. Transcripts of Notch 1 and Jagged 2 in the rat testis were positive throughout the examined period; these intensities became higher at day 13 after birth, coincidence with the formation of spermatocytes, and peaked at day 19 after birth. Expressions of Notch 1 and Jagged 2 were recognized at first in the perinuclear regions of spermatocytes in the rat testis as a round structure at day 19 after birth and thereafter in further differentiated germ cells as meiosis proceeded. In the adult rat testis, positive staining was present as a round structure in spermatocytes, as a typical horseshoe‐shaped structure in round spermatids, and as a covering structure spreading around the nucleus of elongated spermatids, but not in spermatozoa. Notch 1 was recognized in the vacuole of the Golgi complex of primary spermatocytes and the acrosome of elongated spermatids with electron microscopy. When rat testicular tissues were cultured with anti‐Notch 1 or anti‐Jagged 2 antibody, round and elongated spermatids decreased after 5 and 7 days of culture, respectively, and disappeared at around 9 and 12 days of culture, respectively, with shrinkage of the diameter of seminiferous tubules. Spermatocytes, however, increased after 11 days of culture. Expressions of both proteins have been detected in the testicular tissues of human fertile controls as in the rat testicular tissues. However, Notch 1 expression has not been detected in testicular tissues of 11 patients with maturation arrest, whereas Jagged 2 expression has been recognized in all of them. In conclusion, the results presented in this study offer the possibility that Notch 1/Jagged 2 signaling system plays an important role for male germ cells to differentiate or at least to survive in the rat testis and fails to express in the testis of spermatogenic maturation arrest patients.
Expression, Action, and Regulation of Transforming Growth Factor Alpha and Epidermal Growth Factor Receptor During Embryonic and Perinatal Rat Testis Development
Authors: CUPP ANDREA S. , SKINNER MICHAEL K.
Published: December 2015

ABSTRACT: The objective of the current study was to extend previous observations and examine the expression pattern and effects of transforming growth factor alpha (TGFα) and epidermal growth factor receptor (EGFR) on embryonic testis morphogenesis and growth. The expression of TGFα was determined after morphological sex determination (seminiferous cord formation at embryonic day 13 [ED13]) through perinatal testis development (postnatal day 5 [PD5]) with a quantitative reverse transcription‐polymerase chain reaction procedure. Expression of messenger RNA (mRNA) for TGFα appeared to be more dynamic during testis development when compared with the expression of mRNA for EGFR. Message for TGFα was reduced at ED16 and PD4, and was elevated at PD0 during testis development. In contrast, EGFR mRNA levels were negligible at ED15 and were elevated constitutively from ED16 through PD5. Immunohistochemistry was conducted at ED14, ED16, ED19, PD0, PD3, and PD5 to localize cellular expression of both TGFα and EGFR. At ED16, positive staining for EGFR was localized to the cords, and by ED19, was mainly in the cords with slight expression in the interstitium. From PD0 to PD5, positive staining for EGFR was detected in the germ, Sertoli, and interstitial cells. Immunohistochemistry for TGFá detected localization at ED14 and ED16 to the Sertoli cells and to specific cells in the interstitium. From ED19 through PD5, TGFá was detected in the Sertoli, germ, and interstitial cells, and in endothelial cells within the interstitium. To determine the effects of TGFá on embryonic testis growth and seminiferous cord formation, ED13 testis organ cultures were treated with sense and antisense TGFá oligonucleotides. Antisense TGFá inhibited testis growth by 25%‐30% in ED13 testis organ cultures when compared with sense oligonucleotide control pairs. To examine the effects of TGFá on perinatal testis growth, PD0 testis cultures were treated with different doses of TGFá.TGFá increased thymidine incorporation into DNA in PD0 testis cultures. Therefore, TGFá appears to have actions on both embryonic and perinatal testis growth. The regulation of TGFá and EGFR mRNA levels were examined using PD0 testis cultures treated with hormones that stimulate testis growth. Follicle‐stimulating hormone (FSH) stimulated (P < .05) and testosterone tended to stimulate (P < .07) mRNA expression of EGFR. Epidermal growth factor stimulation of PD0 testis cultures did not affect levels of mRNA expression for EGFR, but did suppress expression of mRNA for TGFα. These results taken together demonstrate that TGFα can act to regulate early embryonic and perinatal testis growth. Furthermore, TGFα and EGFR expression can be regulated through growth stimulatory hormones such as FSH and testosterone.
A Flow Cytometric Method for Rapid Determination of Sperm Concentration and Viability in Mammalian and Avian Semen
Authors: Christensen Preben , Stenvang Jens P. , Godfrey William L.
Published: December 2015

ABSTRACT: A new flow cytometric method has been developed to rapidly determine sperm concentration and viability in semen from bulls and boars. Sperm viability was determined on the basis of staining with SYBR‐14 and propidium iodide (PI), and this allowed detection of live (membrane‐intact) sperm, dying (moribund) sperm, as well as dead cells. Fluorescent microspheres (beads) were used to determine sperm concentration. The use of SYBR‐14 at 50 nM and PI at 12 μM in combination with the FACSCount diluent in the counting tubes resulted in a uniform staining after 2.5–5 minutes at room temperature. Reagent staining was reproducible enough to allow subsequent semiautomated analysis of data using Attractors software. In experiment 1, this method was used to analyze semen from boars, rams, rats, rabbits, humans, and turkeys. In experiment 2, Attractors analysis was performed by the FACSCount AF flow cytometer, and sperm concentration determination with this system was compared with results obtained by a spectrophotometer and an electronic cell counter, which is routinely used by bull artificial insemination centers. When compared to microscopic counting in a hemocytometer, the FACSCount AF flow cytometer was two and four times more accurate than the spectrophotometer and the electronic cell counter, respectively. In addition, the FACSCount AF flow cytometer determined both sperm concentration (coefficient of variation 3.3%) and sperm viability (coefficient of variation 0.7%) with high precision.
Rapid Glucocorticoid Mediation of Suppressed Testosterone Biosynthesis in Male Mice Subjected to Immobilization Stress
Authors: Dong Qiang , Salva Antonio , Sottas Chantal M. , Niu Enmei , Holmes Michael , Hardy Matthew P.
Published: December 2015

ABSTRACT: Physical and psychosocial stress challenge homeostasis, increasing glucocorticoid secretion (in rodents, corticosterone [CORT]) while decreasing testosterone (T) levels. The dynamics of stress‐induced changes in T, CORT, and luteinizing hormone (LH) concentrations in mice have not been investigated previously. In particular, it remains to be established whether there is a rapid effect of CORT that is directly mediated by glucocorticoid receptors (GRs) in the testis. Therefore, serum and intratesticular T, serum CORT, and LH levels were measured during acute immobilization (IMO) stress, using the C57BL/6 strain of mice. The effects of testicular GR blockade were investigated by administration of the GR antagonist, RU486, via intratesticular (IT) or intraperitoneal (IP) injection. CORT levels increased in stressed males starting at 15 minutes, reaching a fivefold higher plateau by 1 hour compared with controls (P < .01). Conversely, starting from 30 minutes on, both serum and intratesticular T levels decreased in stressed males to 30% and 8% of control values, respectively, by 6 hours (P < .01). In contrast, LH was unchanged by IMO stress for up to 6 hours. Intratesticular treatment with RU486 partially prevented the IMO‐induced decline in T levels. CORT treatment reduced intracellular cyclic adenosine monophosphate (cAMP) content in Leydig cells by 15 minutes and T production by 30 minutes in vitro. We conclude that 1) the rapid changes in T suggest a suppression of T biosynthesis by glucocorticoid through a nongenomic mechanism, lowering the production of cytoplasmic cAMP; 2) changes in gonadotropic stimulation of Leydig cells are unlikely to explain the suppression of T levels during acute stress; and 3) the results are consistent with a direct inhibitory action of CORT on Leydig cells.
The Utility of Optical Loupe Magnification for Testis Sperm Extraction in Men With Nonobstructive Azoospermia
Authors: Mulhall John P. , Ghaly Sameh W. , Aviv Nadid , Ahmed Absaar
Published: December 2015

ABSTRACT: The testis of patients with nonobstructive azoospermia (NOA) harbors sperm in approximately 30% to 60% of cases. Use of an operating microscope has been shown to result in better sperm retrieval rates. This investigation was undertaken to evaluate the ability of a modified microsurgical approach using magnifying loupes (3.5×) to improve the rates of sperm retrieval during testis sperm extraction (TESE). The study group consisted of patients with NOA who underwent TESE. Before December 1998, TESE was conducted in a standard fashion, and from 1999 on, loupe magnification was used. Comparison was made between the 2 groups with regard to sperm retrieval rates, need for bilateral TESE, and number of tunical incisions. Overall sperm retrieval rates did not differ between the 2 groups (45% vs 50%). However, in patients with testicular volumes of 10 mL or less, patients who underwent standard TESE had a retrieval rate of 27% compared with 42% when using the optical loupe magnification (P = .025). The use of loupe magnification may permit surgeons without access to or experience using an operating microscope to obtain better rates of sperm retrieval in men with NOA who have testicular volumes of 10 mL or less.
Mortality After Atypical Femoral Fractures: A Cohort Study
Authors: Kharazmi Mohammad , Hallberg Pär , Schilcher Jörg , Aspenberg Per , Michaëlsson Karl
Published: December 2015

Although osteoporotic fracture rates can be reduced by bisphosphonates, prolonged therapy is associated with higher risk of atypical femoral fractures. Ordinary fragility fractures are linked to high mortality rates. We aimed to determine whether atypical femoral fractures also confer excess mortality. Radiographs were reviewed for all patients aged ≥55 years who had experienced a subtrochanteric or femoral shaft fracture in Sweden in 2008 to 2010. The fractures were classified as either atypical or ordinary. Data on medication use, coexisting conditions, and date of death were obtained from national registers. We estimated multivariable-adjusted relative risks of death after atypical femoral fractures compared with ordinary subtrochanteric or femoral shaft fractures and calculated age- and sex-standardized mortality ratios (SMRs) for atypical and ordinary fractures compared with the population average. During a mean of 4 years of follow-up, 39 of 172 (23%) patients with an atypical fracture had died compared with 588 of 952 (62%) with an ordinary fracture, corresponding to a relative risk of 0.51 (95% confidence interval [CI] 0.38–0.68). The lower risk was evident in both users and nonusers of bisphosphonates. No patient with atypical fracture died in the first year after fracture. Individuals with an ordinary fracture had a higher mortality risk than the general population (SMR = 1.82; 95% CI 1.69–1.99), but no excess risk was found in patients with atypical fracture (SMR = 0.92; 95% CI 0.65–1.26). We conclude that in contrast to ordinary subtrochanteric and femoral shaft fractures, atypical femoral fractures are not associated with excess mortality. © 2015 American Society for Bone and Mineral Research.
Estimation of the Oxidative Stress and Molecular Damage Caused by 1‐Butyl‐3‐Methylimidazolium Bromide Ionic Liquid in Zebrafish Livers
Authors: Dong Miao , Liu Tong , Wang Jinhua , Wang Jun , Zhu Lusheng , Zhang Jun
Published: December 2015

The present study investigated the toxic effects of 1‐butyl‐3‐methylimidazolium bromide ([C4mim]Br) in zebrafish livers after exposure to 5–40 mg/L of [C4mim]Br on days 7, 14, 21, and 28. The results showed that low levels of [C4mim]Br exposure activated the superoxide dismutase (SOD) activity during early periods of exposure. However, high concentrations (20–40 mg/L) of [C4mim]Br caused the inhibition of SOD, which, accordingly, led to excess reactive oxygen species and malondialdehyde. The present results indicate that [C4mim]Br causes oxidative stress in the livers of both male and female zebrafish. Additionally, the DNA damage revealed that [C4mim]Br has a genotoxic effect on zebrafish livers, even when the concentration of [C4mim]Br is low (5 mg/L). The DNA damage was demonstrated by quantifying the DNA strand breakage.
The Misvaluation Curse in Mergers and Acquisitions
Authors: Pereiro Luis E.
Published: December 2015

When corporate acquirers become self‐mystified by an overly optimistic picture of potential merger gains, they end up trapped in high‐sided valuations, overbid, and eventually overpay for targets—and so destroy their shareholder's value. But there is a flip side too: pessimistic acquirers who systematically underbid end up consistently bypassing potentially profitable acquisition deals. Both faces of misvaluation are rooted in cognitive biases which, being reflex rather than volitional, are difficult—but not impossible—to curb. We suggest four strategies to dilute the misvaluation curse. © 2016 Wiley Periodicals, Inc.
M&A Due Diligence: Consideration of Fraudulent Financial Statements
Authors: McGee Jo Ann , Byington J. Ralph
Published: December 2015

Merger and acquisition (M&A) deals help companies grow quickly (Whitchelo, 2015). However, these types of deals do not come without risks. Corporate management should be aware that when they are conducting M&A due diligence, they should exercise caution in regards to the existence of fraudulent financial statements (FFSs) because of the severe consequences related to FFS. The purpose of this article is to familiarize corporate management with the common fraud schemes employed to create FFS and the red flags that signal the potential existence of these schemes. © 2016 Wiley Periodicals, Inc.
Five Key Reasons Enterprise Data Governance Matters to Finance … and Seven Best Practices to Get You There
Authors: Demarquet Gilles
Published: December 2015

Trends from a recent survey show that enterprise data governance initiatives are more predominant in 2015 than they were in the past. This change is due to CFOs realizing that strong governance around hierarchies and dimensions are critical for enterprise success. Many organizations are planning to reduce by half their use of spreadsheets and e‐mail for enterprise data governance in 2015, and this reduction is matched by an increase in the planned use of packaged enterprise data governance solutions. There are seven best practices to work on to attain strong data governance in your organization. Some may be facilitated through the use of software, but many are just good practices that must be adopted and adhered to by your company. © 2016 Wiley Periodicals, Inc.
Evaluating Materiality in Cybercrime Footnotes
Authors: Smith G. Stevenson
Published: December 2015

New risks face companies dependent on the Internet for their core business processes. New risks come from cyber breaches and attacks from hackers. The results of these challenges are requirements for new footnote disclosures. The article reviews the manner in which those risks have been reported in financial statements based on the materiality of the event. Disclosures for identity theft breaches and bank theft attacks are considered. Questions are raised as to whether intangible losses need to be considered in determining the materiality of a cyber incident as well as suggestions as to how those losses can be estimated. © 2016 Wiley Periodicals, Inc.
The recurrence of negatively reinforced responding of humans
Authors: Alessandri Jérôme , Lattal Kennon A. , Cançado Carlos R.X.
Published: December 2015

The recurrence of negatively reinforced responding of humans was studied in three experiments. In each experiment during Baseline, key‐pressing produced 3‐s timeouts from a requirement to exert finger pressure on a force cell according to variable‐ or fixed‐ratio schedules of reinforcement. In Experiment 1, resurgence was studied by arranging a differential‐reinforcement‐of‐other‐behavior schedule in the second phase, and extinction in the Test phase. In Experiment 2, ABA renewal was studied by extinguishing responding in the second phase in a different context and, in the Test phase, by presenting the Baseline‐phase context when extinction still was in effect. In Experiment 3, reinstatement was studied by arranging extinction in the second phase, followed by the delivery of response‐independent timeouts in the Test phase. Resurgence and renewal occurred consistently for each participant in Experiments 1 and 2, respectively. In Experiment 3, reinstatement was observed less consistently in four participants. The results of these experiments replicate and extend to negatively reinforced responding previous findings of the resurgence and renewal of positively reinforced responding obtained mainly with nonhuman animals.
The occurrence of spermatozoa in the ovary of the gynogenetic viviparous teleost Poeciliaformosa (POECILIIDAE)
Authors: Uribe Mari Carmen , Grier Harry J. , De la Rosa‐Cruz Gabino , Schartl Manfred
Published: December 2015

The reproductive mode of the female viviparous teleost Poecilia formosa (Poeciliidae) represents the phenomenon known as gynogenesis; that is, parthenogenetic development is initiated by spermatozoa which are needed for physiological activation of the egg and the initiation of gestation, but spermatozoa are prevented from contributing to the genome of the embryo. For the reason that no previous histological analyses of the ovary of this species during the reproductive cycle has been published the present study has been conducted. This study examined the histology of the ovary of P. formosa during nongestation and gestation phases and identified the presence of spermatozoa inside the ovary. Spermatozoa were observed in folds of the ovarian epithelium of P. formosa during both the nongestation and gestation phases. Sperm storage as documented in this study is a very important trait for the gynogenetic viviparous fish P. formosa contributing to the understanding of this species reproduction. J. Morphol. 277:341–350, 2016. © 2015 Wiley Periodicals, Inc.
Analysis of frequency and magnitude of extreme rainfall events with potential impacts on flooding: a case study from the central United States
Authors: Rahmani Vahid , Hutchinson Stacy L. , Harrington John A. , Hutchinson J. M. Shawn
Published: December 2015

Climate variability and change can impact rainfall by varying time, location, magnitude, and frequency of precipitation events. Fluctuations in heavy rainfall events can impact flooding and drought events and water management systems. This research addresses temporal and spatial distributions of extreme daily and monthly rainfall in Kansas using daily rainfall data from 23 stations for the period 1890–2013. The Mann–Kendall non‐parametric method was used in trend analysis. Results indicate an increasing trend in the annual daily maximum rainfall, and an increase in the annual number of rainfall events above the 90th, 95th, and 99th percentile thresholds at a majority of stations since 1890. The most recent 30‐year climate normal period (1981–2010) was selected to assess contemporary change compared to the entire period (1890–2013). Most stations have a steeper positive slope for all extreme rainfall parameters for 1981–2010. Generally, western Kansas receives smaller and fewer extreme storms than eastern Kansas with respect to both magnitude and frequency.

Since 1890, June has been the month that receives the greatest amount of rain in each year at a majority of stations (18 of 23). Stations in eastern Kansas tend to experience earlier maximum rainfall on a monthly basis than central and western stations. Earlier annual maximum monthly rains can affect soil moisture and runoff generation. Timely maintenance and improvement in water, soil, and flood management systems are necessary in order to increase preparedness of the society to flash floods and protect them from water management systems failures.
Labile organic matter fractions as early‐season nitrogen supply indicators in manure‐amended soils
Authors: Thomas Ben W. , Whalen Joann K. , Sharifi Mehdi , Chantigny Martin , Zebarth Bernie J.
Published: December 2015

Soil test indicators are needed to predict the contribution of soil organic N to crop N requirements. Labile organic matter (OM) fractions containing C and N are readily metabolized by soil microorganisms, which leads to N mineralization and contributes to the soil N supply to crops. The objective of this study was to identify labile OM fractions that could be indicators of the soil N supply by evaluating the relationship between the soil N supply, the C and N concentrations, and C/N ratios of water extractable OM, hot‐water extractable OM, particulate OM, microbial biomass, and salt extractable OM. Labile OM fractions were measured before planting spring wheat (Triticum aestivum L.) in fertilized soils and the soil N supply was determined from the wheat N uptake and soil mineral N concentration after 6 weeks. Prior to the study, fertilized sandy loam and silty clay soils received three annual applications of 90 kg available N (ha · y)−1 from mineral fertilizer, liquid dairy cattle manure, liquid swine manure or solid poultry litter, and there was a zero‐N control. Water extractable organic N was the only labile OM fraction to be affected by fertilization in both soil types (P < 0.01). Across both test soils, the soil N supply was significantly correlated with the particulate OM N (r = 0.87, P < 0.001), the particulate OM C (r = 0.83, P < 0.001), and hot‐water extractable organic N (r = 0.81, P < 0.001). We conclude that pre‐planting concentrations of particulate OM and hot‐water extractable organic N could be early season indicators of the soil N supply in fertilized soils of the Saint Lawrence River Lowlands in Quebec, Canada. The suitability of these pre‐planting indicators to predict the soil N supply under field conditions and in fertilized soils from other regions remains to be determined.
Quality control of fragrances using Raman spectroscopy and multivariate analysis
Authors: Godinho Robson B. , Santos Mauricio C. , Poppi Ronei J.
Published: December 2015

An analytical methodology using Raman spectroscopy and chemometrics was developed for direct, fast and non‐destructive discrimination and prediction of the properties of fragrances according to their composition. The soft independent modeling of class analogies was used as a supervised classification method for fragrances classification, and partial least squares regression as a multivariate calibration method for the prediction of physicochemical properties of fragrances, such as density and refractive index. From 155 fragrance samples, the model exhibited a high success rate for all of the studied fragrance classes, with 100% correct classification. In the multivariate calibration model, adequate correlation was observed between the measured and partial least squares regression‐predicted data for refractive index and density, with a relative standard error of prediction between 0.02% and 0.07%, respectively. This study demonstrates the wide applicability of the methodology for the discrimination, classification, and prediction of complex olfactory mixtures in quality control of fragrances. Copyright © 2015 John Wiley & Sons, Ltd.
Antihyperlipidemic effect of Acanthopanax senticosus (Rupr. et Maxim) Harms leaves in high‐fat‐diet fed mice
Authors: Nishida Miyako , Kondo Momoko , Shimizu Taro , Saito Tetsuo , Sato Shinji , Hirayama Masao , Konishi Tetsuya , Nishida Hiroshi
Published: December 2015

BACKGROUND

Metabolic syndrome is a major risk factor for a variety of obesity‐related diseases. Recently, the effects of functional foods have been investigated on lipid metabolism as a means to reduce lipid content in the blood, liver and adipose tissues associated with carnitine O‐palmitoyltransferase (CPT) activity. Acanthopanax senticosus (Rupr. et Maxim) Harms (AS) is a medicinal herb possessing a wide spectra of functions including antioxidant, anti‐inflammatory and anti‐fatigue actions. Despite much research being focused on the cortical roots of AS, little information is available regarding its leaves, which are also expected to promote human health, for example by improving abnormal lipid metabolism. Here, we explored whether AS leaves affect lipid metabolism in mice fed a high‐fat diet.

RESULTS

The administration of AS to BALB/c mice fed a high‐fat diet significantly decreased plasma triglycerides (TG). CPT activity in the liver of these mice was significantly enhanced by AS treatment.

CONCLUSION

These findings indicate that AS leaves have the potential to alleviate increase in plasma TG levels due to high‐fat diet intake in mice, possibly by increasing mitochondrial fatty acid β‐oxidation, especially via CPT activation. Consequently, daily intake of AS leaves could promote beneficial health effects including the prevention of metabolic syndrome. © 2015 Society of Chemical Industry