Effect of methylene blue, indigo carmine, and Renografin on human sperm motility

OBJECTIVES: To evaluate the effect of commonly used intraoperative vasography and tissue staining agents, indigo carmine, methylene blue, and Renografin, on sperm motility. METHODS: Semen from 20 healthy men was obtained after 2 to 4 days of abstinence. Sperm motility was initially evaluated in each specimen. Standard solutions of indigo carmine, methylene blue, and Renografin-60 were diluted 2x and 4x with lactated Ringer's solution. Equal aliquots of sperm were mixed with undiluted and diluted drugs, and sperm motility was assessed. RESULTS: Initial mean sperm motility was 70.3%+/-3.0%. Undiluted methylene blue and Renografin severely depressed sperm motility to 1.1%+/-0.5% and 2.3%+/-0.7%, respectively (P <0.05). Diluted methylene blue depressed motility to 4.9%+/-1.8% and 11.2%+/-3.0% (P < 0.05). Diluted Renografin depressed motility to 25.1%+/-4.1% and 55.3%+/-3.3% (P < 0.05). Although undiluted and 2x-diluted indigo carmine moderately decreased sperm motility (48.9%+/-3.2% and 61.7%+/-3.0%, P < 0.05), 4x-diluted indigo carmine had minimal effect on sperm motility (67.3%+/-2.8%, P > 0.05). Lactated Ringer's solution had no effect on sperm motility. CONCLUSIONS: We found a severe, immediate reduction in sperm motility after exposure to undiluted standard solutions of methylene blue and Renografin. Dilution of Renografin significantly decreased its negative impact on the sperm motility, whereas the adverse effect of methylene blue remained fairly constant even with increasing dilution. Sperm motility should be assessed prior to application of these agents. Sperm should be aspirated for immediate use and/or cryopreservation prior to the use of these agents. Indigo carmine may be safely used as a tissue stain or vasography agent with a minimal effect on sperm motility in dilutions of 4x and higher.     

Effect of the phosphodiesterase inhibitor Pentoxyfylline on human sperm motility

Tyrode fluid and Tyrode fluid plus Pentoxifylline were individually added to aliquots of semen samples obtained from 6 normal men and 6 infertile patients considered to have idiopathic normogonadotropic oligoasthenozoospermia. Pentoxifylline was added to final concentrations of 0.15, 0.30 and 0.60 mM. One aliquot with no addition served as control. Samples were incubated in 37 degrees C and observed by light microscopy at 30 minutes and at 1, 2 and 4 hours after obtaining the material. At observation time, semen quality was evaluated by determining the percentages of forwardly progressive spermatozoa, slowly progressive spermatozoa, "in situ" motile spermatozoa, live and non-motile spermatozoa and dead spermatozoa. Results reported included only the first and last category. Tyrode fluid did not affect significantly the motility and the duration of activity of spermatozoa. Ejaculated human spermatozoa both from normal and asthenozoospermic men added the Pentoxifylline at 0.30 and 0.60 mM showed a longer lasting activity than those of control semen and semen added only with Tyrode fluid.     

Effects of trehalose and sucrose, osmolality of the freezing medium, and cooling rate on viability and intactness of bull sperm after freezing and thawing

The effect of passively exposing infants to visual information in a sensory preconditioning paradigm was assessed in five experiments with seventy-eight 6-month-olds. In the basic paradigm, infants were simultaneously exposed to two contexts (S1 and S2), trained in one of them (S1), and tested in the other (S2). Infants learned an association between S1 and S2 and exhibited transfer to S2 after preexposures of 1 hr/day for 1 week but not after 2 min or 1 hr on a single occasion. When S1 was the focal cue, however, infants transferred responding after a single preexposure lasting only 2 min. Extended preexposure also produced learned irrelevance, biasing subsequent selective attention, but there was no evidence of latent inhibition. These findings demonstrate that young infants pick up fairly sophisticated information about specific stimuli in their visual surround and the relationships between them merely as a result of passive observation, without explicitly being trained to do so. Moreover, infants may not express this knowledge for several days, until finally given an opportunity to do so. Sensory preconditioning may be an instance of "unitization"-a special type of infantile learning that enables immature organisms to acquire some associations more rapidly than adults.     

Effect of sodium nitroprusside on sperm motility, viability, and lipid peroxidation

OBJECTIVE: To analyze the effect of sodium nitroprusside, a nitric oxide releaser, on sperm motion and lipid peroxidation-induced membrane damage in cryopreserved human sperm. DESIGN: Post-thaw, cryopreserved, human sperm samples were washed and divided into three aliquots. Each aliquot was incubated with either 0, 50, or 100 nM sodium nitroprusside. INTERVENTIONS: Samples were analyzed for lipid peroxidation (measured by malonaldehyde-thiobarbituric acid reactivity) at 3 hours post-thaw. MAIN OUTCOME MEASURES: Percent viability and motion parameters were assessed at 0, 10, and 30 minutes and 2, 3, 5, and 6 hours post-thaw. RESULTS: All results represent a mean +/- SEM, n = 10. Lipid peroxidation in samples incubated with 50 nM sodium nitroprusside (15.1 +/- 2.1 nM malonaldehyde/10(8) sperm) or 100 nM sodium nitroprusside (13.2 +/- 2.1 nM malonaldehyde/10(8) sperm) was significantly lower than in controls (22.7 +/- 3.1 nM malonaldehyde/10(8) sperm). Percent viability was significantly reduced from 0 minutes (60.6% +/- 3.5%) to 6 hours post-thaw in controls (38.0% +/- 5.1%) but not in 50 nM (46.8% +/- 10.4%) or 100 nM (48.8% +/- 6.5%) sodium nitroprusside-treated samples. Compared with controls (18.3% +/- 3.4%), maintenance of percent motility at 3 hours post-thaw was significantly improved in 50 nM (24.5% +/- 2.9%) and in 100 nM (26.3% +/- 3.2%) sodium nitroprusside-treated samples. Straight line velocity maintenance was significantly improved in 50 nM (37.3 +/- 1.3) and in 100 nM (37.0 +/- 1) sodium nitroprusside-treated samples as compared with controls (30.5 +/- 1.7). Significant improvements in curvilinear velocity maintenance compared with controls (56.3 +/- 2.9) also were observed in 50 nM (65.9 +/- 2.1) and 100 nM (72.1 +/- 4.1) sodium nitroprusside-treated samples. Significant differences in the motion parameters of sodium nitroprusside-treated samples were maintained at 5 and 6 hours post-thaw in comparison to controls. CONCLUSION: These results suggest that sodium nitroprusside is beneficial to the maintenance of post-thaw sperm motion and viability for up to 6 hours and that reduction of lipid peroxidative damage to sperm membranes may be the mechanism for these benefits.     

Effects of alprostadil and prazosin on motility, viability and membrane integrity of human sperm

PURPOSE: We evaluated the effects of alprostadil, prazosin hydrochloride, and alprostadil/prazosin hydrochloride, agents used in the clinical treatment of male erectile dysfunction, on the motility, viability and membrane integrity of human sperm. MATERIALS AND METHODS: Ten healthy volunteers provided semen samples that were incubated with 0.4 mg./ml. alprostadil, 0.1 and 0.2 mg./ml. prazosin hydrochloride and 0.4 mg./ml. alprostadil plus 0.1 mg./ml. prazosin hydrochloride for 2 hours. Control incubations included polyethylene glycol 1450, the formulation vehicle for the clinical use of alprostadil and prazosin, and Ham's F-10 buffer. Serial evaluations of percent sperm motility, percent viability, membrane function (by hypo-osmotic swelling test) and several computer generated measurements of sperm motion, including straight line velocity, curvilinear velocity, linearity and amplitude of lateral head displacement, were made. RESULTS: None of the agents had a significant impact on the percentage of motile or viable sperm or on sperm membrane function. Incubation with 0.2 mg./ml. prazosin reduced straight line velocity and curvilinear velocity significantly compared with the other agents. These changes were most likely a direct result of the viscosity of the 0.2 mg./ml. prazosin solution and not a cellular or metabolic effect on the sperm. CONCLUSIONS: Alprostadil and prazosin hydrochloride at doses used in transurethral therapy for erectile dysfunction have no effect on the motility, viability and membrane integrity of human sperm.     

Effect of angiotensin converting enzyme (ACE) and angiotensins on human sperm functions

The presence of components of the renin angiotensin system (RAS) and specific receptors of angiotensin II in the female and male reproductive tract supports the hypothesis that reproductive functions may be controlled by RAS. Therefore, the present study investigated the influence of ACE and angiotensins on sperm functions and the sperm-egg interaction. The experiments did not indicate direct effects of ACE on the capacitation process or acrosome reaction. Release of ACE from human spermatozoa during capacitation was not related to their ability to undergo acrosome reaction after stimulation with ionophore. Therefore, ACE release does not seem to be a useful clinical marker for human sperm capacitation. However, decreased binding of human spermatozoa to the oolemma of zonafree hamster oocytes after inhibition of ACE by captopril indicates that kininase II is involved in sperm-egg interactions. In contrast to other studies, incubation with captopril had no influence on sperm binding to the zona pellucida. Because effects of ACE on sperm-egg interactions but not on capacitation or acrosome reaction were observed, several experiments were performed to study the influence of substrates and products on the acrosome reaction. Angiotensin II induced the acrosome reaction dose-dependently, whereas angiotensin I had no effect on the acrosome reaction. The effect of angiotensin II on acrosome reaction seems to be calcium-dependent and mediated by protein kinases. Since a specific type 2 angiotensin II receptor inhibits the acrosome reaction induced by angiotensin II, this subtype of receptors may be present at the surface of sperm heads. Another clue for the presence of type 2 receptors on human spermatozoa is the finding that pertussis toxin did not inhibit the angiotensin II induced acrosome reaction. In contrast to type 1 angiotensin II receptors, type 2 receptors are known to be G-protein independent.     

Effect of temperature and viscosity on the motility of the spirochete Treponema denticola

Two adult siblings with early onset dementia are described. At presentation, in their early 30s, they showed poor judgment and disinhibition. A progressive dementia ensued over several years. Brain MRI disclosed diffusely increased T2 signal in the cerebral white matter, suggestive of a leukodystrophy. Numerous lysosomal enzyme assays including leucocyte arylsulphatase A and galactocerebrosidase activities, plasma and fibroblast very long chain fatty acid concentrations, and urinary sulphatide concentrations were normal, as were CSF analyses. A brain biopsy disclosed periodic acid Schiff (PAS) and Sudan black positive material in perivascular macrophages which, by electron microscopy, consisted of stacks of straight or curvilinear paired membranes within angulate lysosomes, indicative of abnormal glycolipid accumulation. The combination of clinical, radiological, biochemical, and pathological features of this degenerative disease is not consistent with that of any of the known leukodystrophies or lysosomal storage disorders. These findings suggest a previously undescribed familial glycolipid storage disorder causing an adult onset leukodystrophy and presenting with behavioural symptoms that mimic a psychiatric disorder.     

Comparison of sperm separation methods: effect on recovery, motility, motion parameters, and hyperactivation

OBJECTIVE: To compare the Enhance (Percoll; Conception Technologies, San Diego, CA) and PureSperm (Gen X International, Madison, CT) sperm preparation methods with respect to recovery (percentage of motile sperm), motility (%), path and progressive velocities (microm/s), and hyperactivation (%). DESIGN: Comparison of sperm processing methods. SETTING: University medical center-based clinical andrology laboratory and infertility program. PATIENT(S): Twenty-five men who presented for semen analysis. INTERVENTION(S): Each of 25 semen specimens were divided and each aliquot was prepared using two different density gradient centrifugation methods. MAIN OUTCOME MEASURE(S): The motile sperm recovery, percent motility, motion parameters, and percent hyperactivation were measured for each semen specimen (n=25) before and after separation with the use of the two methods. RESULT(S): There was no difference in the percent motility and motile count between specimens prepared with Enhance (Percoll) and PureSperm and fresh specimens. Statistically significant differences were found (fresh versus test) in the velocities and in hyperactivation (PureSperm only), and no differences were found between the processing methods. CONCLUSION(S): PureSperm appears to be as effective as Percoll (Enhance) for the recovery of good, progressively motile sperm for use in IUI or other assisted reproductive techniques.     

Differential effects of mucosal pH on human (Caco-2) intestinal epithelial cell motility, proliferation, and differentiation

Mucosal pH abnormalities are associated with anastomotic dehiscence, ischemia, and malignancy. We postulated that intraluminal pH influences intestinal epithelial motility, proliferation, and differentiation and studied extracellular pHo (7.0-8.5) effects on human (Caco-2) intestinal epithelial motility, proliferation, and differentiation. Mucosal healing was modeled by sheet migration and differentiation by alkaline phosphatase and dipeptidyl dipeptidase specific activity. In parallel differentiation and motility studies, we inhibited proliferation with mitomycin to dissociate indirect mitogenic effects. Intracellular pHi was quantitated using BCECF/AM at varying extracellular pHo and in migrating cells. Motility was maximal at pHo 7.6 and proliferation at 7.2. Each decreased with acidity and alkalinity. By contrast, brush border enzyme activity was lowest at pHo 7.0 and highest at pHo 8.5. pHi was highest at pHo 8.5. Migrating cell pHi was higher than static cell pHi. Thus, extracellular pHo deviations perturb Caco-2 pHi homeostasis and motility. Alkalinity promotes differentiation while acidity induces proliferation and limits differentiation.     

The relationship between sperm morphology and rates of fertilization, pregnancy and spontaneous abortion in an in-vitro fertilization/intracytoplasmic sperm injection programme

The morphological normality of a spermatozoon is considered to be an important factor in relation to its ability to fertilize an oocyte. We examined the influence of morphology (strict criteria) on the rates of fertilization, pregnancy and spontaneous abortion obtained following conventional in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in our clinical programme. We found our fertilization cut-off values for conventional IVF to be slightly different from those of the Kruger group (10 and 5%, compared to 14 and 5%). We also found the pregnancy rate per transfer to be as good or better in the groups with < 5% normal forms: 36% of these men had a fertilization rate > 50% using conventional IVF, showing that fertilization capacity is not necessarily impaired even in this 'poor prognosis' group. With the exception of the ICSI group with 5-9% normal forms, the rate of spontaneous abortion in this study was similar to or lower than in our IVF/ICSI programme overall. When the 5-9% normal spermatozoa group was divided into those with teratozoospermia as the only factor and those with additional sperm factors, the increased abortion rate was found in the group with multiple sperm factors (67% spontaneous abortions).     

Nb对V、N微合金化压力容器钢珠光体形态及低温韧性影响

为了在保证强度同时提升压力容器钢低温韧性,以铌(Nb)取代部分钒(V),研究Nb的添加对V、N微合金化正火压力容器钢板显微组织和力学性能的影响。结果表明,Nb+V试验钢相比于V试验钢,在强度保持630 MPa的同时可显著提升低温冲击韧性,-20、-40、-60℃冲击吸收功分别提升59、44、29 J。原因主要为Nb的添加促进了细小铁素体转变,其含量的增加使钢的塑韧性增强。此外V和Nb+V钢中碳原子扩散系数在平均转变温度下分别为2.64×10^-13、2.14×10^-13 cm²/s, Nb的添加降低了珠光体生长速度、增加了珠光体转变过程中的过冷度,从而细化了珠光体团簇和片层间距。EBSD分析可知以Nb取代部分V后试验钢有效晶粒尺寸由7.7μm减小至5.8μm,大角度晶界比例由83.1%增加至85.5%。微合金元素Nb的细晶作用及对大角度晶界比例提升的贡献也可以大幅提高钢的低温冲击韧性。     

超细低氧Ti(CN)粉末的制备及合成温度对其性能与结构的影响

以超细TiO2、碳黑和高纯氮气为原料,在高温下直接合成了超细Ti(C1-xNx)粉末.结果显示,通过C/Ti的准确配比,适当的反应温度、N2流速和保温时间及高效球磨工艺可以制得低氧含量的单相Ti(C1-xNx)超细粉末.经高效球磨破碎后,可得到费氏粒度为0.4μm、晶粒度为40nm的超细Ti(CN)粉末.Ti(C1-xNx)的点阵常数与x值有很好的对应关系,并且随着反应温度的升高及保温时间的延长,Ti(C1-xNx)粉末x值降低,粒度和晶粒度增大,.     

合金成分对高温Sm(Co,Fe,Cu,Zr)z磁性能与显微组织的影响

用粉末冶金法制备了6种Sm(CobalFe0.1CuyZr0.04)z烧结磁体,每种成分磁体都进行了热处理工艺优化实验;并对合金成分对磁体的显微组织及磁性能的影响进行了研究.结果表明z值较小的磁体有较多的15相,室温的Br、Hα与(BH)max都较低些,但方形度(K)较好,因此适当减小z值有利于提高高温磁性能;Cu含量的增加使15相的畴壁能(γ5)减少,217相与15相的畴壁能差(△γ)增加,从而使矫顽力增加,但Cu含量太高,如果不同时增加Sm含量,将导致Br与(BH)max急剧下降;z值较低,同时Cu含量又较高的Sm(CobalFe0.1-Cu0.16Zr0.04)6.7磁体具有最佳室温磁性能与方形度.     

温度对碳热还原合成Ti(C,N)晶须的影响

以TiO2和碳黑为主要原料,NaCl为卤化剂,NiCl2粉作催化剂,N2为保护气氛,采用碳热还原法,在不同温度下合成Ti(C,N)晶须.用X-Ray衍射仪研究合成温度对样品的相组成的影响;采用扫描电镜观察样品的形貌,并检测粉末中的残留碳、氧含量.结果表明:合成温度对样品的相组成、形貌及游离碳和氧含量有重要影响.在1 200℃下主要发生TiO2与碳黑、氮气间的直接还原反应形成TiN,粉末中游离碳和氧含量分别为22.54%和19.82%,合成粉末由大量颗粒组成;温度升高至1 400℃时,粉末由Ti(C,N)和Ni两相组成,游离碳和氧含量分别降低至0.57%和0.38%,样品成为少量的颗粒与大量晶须组成的混合物,晶须长度在15～30 μm之间,直径约1 μm;晶须表面光滑.     

Influence of co-culture with established human endometrial epithelial and stromal cell lines on sperm movement characteristics

The effects of co-culture of human spermatozoa with human immortalized endometrial cells - epithelial or stromal - on sperm movement characteristics, including hyperactivation, were studied using computer-assisted sperm analysis (CASA). Epithelial and stromal cell types could be separated following 8-10 days of culture of endometrial cells originating from human biopsies. Both cell types were immortalized by the SV 40 large T antigen. Co-incubation of sperm with epithelial and stromal monolayers enhanced the rate of hyperactivation: 24.9% (P <0.05) and 17.8% (P = 0.05) versus 9.5% as control, respectively, whereas the majority of motility parameters remained unchanged. Conditioned media had no effect upon sperm parameters, including hyperactivation. Co-incubation with either monolayer was able to maintain sperm motility over a longer period than incubation in control medium alone. In four patients whose spermatozoa did not exhibit hyperactivation, co-incubation with epithelial cells, but not conditioned medium, allowed normal rates of hyperactivation (range: 6.9-15.6%).     

Effect of sintering temperature on morphology and arc erosion properties of La-Ni-O ceramic and its composites

Ceramic LaNiO3 samples were prepared by solid state reaction method at different sintering temperatures. It was found that the resultant was not ABO3 perovskite single phase but dual phase La2NiO4 and NiO, and the percentage of the two phases varied with sintering temperature. Ceramics sintered at 1400 ℃ were well crystallized and the phase ratio of La2NiO4 was the maximum. The surface morphology observed by scanning electron microscopy (SEM) indicated that the grains of the ceramics sintered at 1400 ℃ were uniform and compact, which were in agreement with the properties of high density and low electrical resistivity of the samples. X-ray diffraction (XRD) patterns of ceramics before and after arc erosion indicated their high structural stability, which resulted in the good arc erosion resistance properties for silver-based electrical contact materials. The contact materials prepared with the ceramic sintered at 1400 ℃ exhibited better mass transition and arc erosion resisting properties.     

Primed in situ labeling (PRINS): a method for rapid identification and quantification of human chromosomes in both lymphocytes and sperm nuclei

In this work, a specific primer for X alphoid satellite DNA was used to detect chromosome X through primed in situ labeling (PRINS). The method allows the rapid identification of chromosome X in metaphase and its quantification in interphase. PRINS is equally applicable to both lymphocytes and sperm nuclei.     

Effect of anti-freeze protein (AFP) on the cooling and freezing of equine embryos as measured by DAPI-staining

Equine embryos recovered on Day 6 after ovulation were cooled to +4 degrees C, or frozen with AFP alone or together with glycerol. Twenty embryos (140-200 microm in diameter) were randomly assigned to 6 treatment groups. In the first 3 groups, the embryos were cooled from room temperature to +4 degrees C at a rate of 3 degrees C/min and warmed again at a rate of 32 degrees C/min in a programmable freezer. In the second 3 groups, the embryos were frozen using a standard protocol, stored in liquid nitrogen for 5-7 days and then thawed in a 37 degrees C waterbath. After cooling/warming or freezing/thawing all the embryos were stained with DAPI. The percentage of dead cell area was significantly lower in the cooling groups than in the freezing groups and no significant differences were apparent between the cryoprotectants used in the study.