Park, Yong-Seog;Lee, Hyoung-Song;Song, Sang-Jin;Kim, Jeong-Wook;Kang, Inn-Soo;Seo, Ju-Tae
Clinical and Experimental Reproductive Medicine
/
v.27
no.3
/
pp.267-273
/
2000
Objectives: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. Materials and Methods: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}C$ water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. Results: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospennic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. Conclusions: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.
Kim, Hyeon-Yeong;Lee, Sung-Bae;Han, Jeong-Hee;Choi, Sung-Bong;Kang, Min-Gu;Lee, Chae-Kwan
Journal of Korean Society of Occupational and Environmental Hygiene
/
v.17
no.3
/
pp.224-234
/
2007
This study aimed to examine the harmful effects of Mn and Fe, which may be generated as dust or fume in the industrial sites, on the body and genital organs by their inhalation. It is intended to find the characteristics and differences of the hazardousness by inhaling a single and the mixed materials of Mn and Fe. Male F344 rats were divided into the control group and 3 exposed groups on the basis of the test material compound (Mn $1.5mg/m^3$, Mn 1.5 and Fe $3.0mg/m^3$, Fe $3.0mg/m^3$). The 4 groups were divided into 4 subgroups again on the basis of the exposure period (4 and 13 weeks) and the recovery period (4 and 13 weeks). The exposure condition was 6 hours a day, 5 days a week for the whole body. Clinical tests including changes in weight and feed rate, blood biochemical test, motility change, changes in the number and the amount of spermatozoon (sperm count), daily sperm production (DSP), deformity test of spermatozoon and changes in the accumulation of Mn and Fe in blood and internal organs were performed. Motility was reduced by Mn exposure. Especially, the effect of Mn was exposure period responsible. By mixing with Fe, no significant change in motility Mn and Fe accumulation in organs was observed. Sperm count and daily sperm production (DSP) were decreased by Mn. Additional effect like the reduction of sperm count and DSP, and delayed restoration of sperm count and DSP during the recovery period were observed in the mixed exposure group. These results indicate that Mn and Fe may affect the motility reduced and has male reproductive toxicity. Mixed exposure of Mn and Fe lead to synergic effects on the male reproductive toxicity.
Proceedings of the Korean Environmental Health Society Conference
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2005.06a
/
pp.319-322
/
2005
Porous building materials are not only sources of indoor air pollutants such as volatile organic compounds (VOCs) but they are also strong sinks of these pollutants. Volatile organic compounds have been implicated in impaired spermatogenesis, increase in the incidence of malformed sperm and decrease in the percentage of moving sperm. The aim of this study was to determine and compare the direct effects of various volatile organic compounds (phenol, formaldehyde; HCHO, ethanol, toluene, styrene) on motility and survival rate of human sperm in vitro. Semen samples from 3 health subjects were prepared using swim-up method and 1-10mM volatile organic compounds were added to the test medium. HCHO and phenol produced significant decreases in the motility and survival rate with a different potency. The most potent inhibition of motility and survival rate was observed after exposure to HCHO. Less than 1mM HCHO significantly inhibited sperm motility. When ethanol is added directly to sperm, at concentrations equivalent to that in serum after heavy drinking, these damaging effects were lowest compared with other volatile organic compounds. Present study shows that each compound has differential toxic potency to human sperm and we need special caution for the use of HCHO and phenol.
Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG $0.2{\mu}M$, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.
The author performed radionuclide esophageal transit studies(RETS) with liquid and solid boluses using the same day protocol in 90 normal controls and 164 patients with various primary esophageal motility disorders who were diagnosed by manometric criteria and clinical courses. The authors calculated mean esophageal transit time(MTT) and mean residual retention(MRR) in each of the liquid and solid studies, and classified time-activity curve(TAC) patterns. The normal criteria of RETS with liquid bolus were MTT<24 sec, MRR<9%, and the TAC pattern that showed rapid declining slope and flat low residual(Type 1). The normal criteria of RETS with solid bolus were MTT<35 sec, MRR<9% and TAC of type 1. With these normal criteria, the sensitivity and the specificity of the liquid study were 62.2 % and 97.8%, respectively. The sensitivity increased to 75.4% with the solid study. The author also found that the RETS was highly reproducible. The achalasia typically showed no effective emptying of both liquid and solid boluses during the whole study period, and was well differentiated by its extremely long transit time and high retention from the other motility disorders. The diffuse esophageal spasm (DES) and nonspecific esophageal motility disorder(NEMD) showed intermediate delay in transit time and increased retention. In the groups of hypertensive lower esophageal sphincter(LES), hypotensive LES and nutcracker, there noted no significant difference with the normal control group in terms of MTT and MRR. The DES and NEMD could be more easily identified by solid studies that showed more marked delay in MTT and increased MRR as compared with the liquid study. In conclusion, esophageal scintigraphy is a safe, noninvasive and physiologic method for the evaluation of esophageal emptying.
Wong, Uni;Person, Erik B;Castell, Donald O;von Rosenvinge, Erik;Raufman, Jean-Pierre;Xie, Guofeng
Journal of Neurogastroenterology and Motility
/
v.24
no.4
/
pp.570-576
/
2018
Background/Aims Swallows with viscous or solid boluses in different body positions alter esophageal manometry patterns. Limitations of previous studies include lack of standardized viscous substrates and the need for chewing prior to swallowing solid boluses. We hypothesize that high-resolution impedance manometry (HRiM) using standardized viscous and super-viscous swallows in supine and upright positions improves sensitivity for detecting esophageal motility abnormalities when compared with traditional saline swallows. To establish normative values for these novel substrates, we recruited healthy volunteers and performed HRiM. Methods Standardized viscous and super-viscous substrates were prepared using "Thick-It" food thickener and a rotational viscometer. All swallows were administered in 5-mL increments in both supine and upright positions. HRiM metrics and impedance (bolus transit) were calculated. We used a paired two-tailed t test to compare all metrics by position and substrate. Results The 5-g, 7-g, and 10-g substrates measured 5000, 36 200, and 64 $700mPa{\cdot}sec$, respectively. In 18 volunteers, we observed that the integrated relaxation pressure was lower when upright than when supine for all substrates (P < 0.01). The 10-g substrate significantly increased integrated relaxation pressure when compared to saline in the supine position (P < 0.01). Substrates and positions also affected distal contractile integral, distal latency, and impedance values. Conclusions We examined HRiM values using novel standardized viscous and super-viscous substrates in healthy subjects for both supine and upright positions. We found that viscosity and position affected HRiM Chicago metrics and have potential to increase the sensitivity of esophageal manometry.
Sara Boushaba;Yassine Helis;Rachida Lebaal;Sabah Beldjebel;Ayache Benhamza;Chafia Ziti;Ghania Belaaloui
Clinical and Experimental Reproductive Medicine
/
v.50
no.1
/
pp.53-62
/
2023
Objective: The aim of this study was to investigate the relationships of serum folate (vitamin B9), cobalamin (vitamin B12) levels and diet with semen parameters (semen standard parameters [SSP] and DNA fragmentation index [DFI]) in infertile men. Methods: Sperm samples were assessed for SSP and DFI (using the sperm chromatin dispersion test). Serum vitamin concentrations were measured with an immuno-electrochemiluminescence assay, and men completed a semi-quantitative food frequency questionnaire (FFQ). Results: Serum folate levels were positively correlated with sperm progressive motility and DFI. A comparison of SSP between two groups of patients according to serum folate concentration (B9 <4.840 ng/mL and B9 ≥4.840 ng/mL) showed significantly higher sperm concentration and sperm progressive motility in the latter group. However, there was no difference between these groups regarding DFI. Interestingly, serum folate levels were significantly higher in patients with a high DFI (using the cut-offs of 30% or 18%). FFQ data showed that the consumption of fruits and egg yolk correlated positively with sperm concentration and sperm motility, respectively. Conclusion: Serum folate levels showed significant associations with sperm concentration and sperm progressive motility. However, the positive association of serum folate with DFI raises the need for careful prescription of folate supplements.
The influences of the acute hemorrhage on the intestinal functions were studied in the rabbits subjected to acute bleeding, amounting 1.5-2% of the body weight. The motility and the absorptive capacity of the ileum were compared before and after the bleeding. Transfusion of shed blood was also performed in order to see whether the deteriorations were reversible or not. The tension developed in the direction of the longitudinal axis of the ileum was recorded through an appropriate transducer, and the frequency of the rhythmic contraction was counted throughout the procedure. Test solution, 10ml in amount, was placed in the loop of the ileum, and the samples were drawn at zero time and at 20 minutes. Triplicated procedures were repeated on the same loop;namely, before and after bleeding and after transfusion. The test solution was composed of 200 mg% urea, 218 mEq/l of NaCl and 150 mg% of polyethylene glycol (PEG) No. 4,000 in distilled water. The latter substance was used as a marker substance for the volume change of the loop. The results obtained were as follows; 1. The motility of the ileum suffered little effects by acute hemorrhage. However, minor fluctuations were seen in the frequency of the rhythm, showing a slight tendency of decreasing rhythmicity, and it was reversed by transfusion. 2. Diminution of absorptive capacity of urea was noticed in acute hemorrhage and it was interpreted as the consequence of the secondary effect of the retardation of the active transport mechanism governing the sodium transport 3. Absorption rate of the sodium ion was dropped in the hemorrhage, suggesting the indispensable need of the blood supply. 4. Osmolarity of the luminal fluid remained higher in the case of acute hemorrhage. 5. There was a tendency of retaining more fluid in the intestinal lumen in acute hemorrhage, comparing with that observed prior to the bleeding. 6. The deteriorations in the absorptive capacity were restored by transfusion of shed blood.
Background/Aims Potassium-competitive acid blockers are expected to be the next generation of drugs for the treatment of diseases caused by gastric acid. In 2015, vonoprazan fumarate, a novel potassium-competitive acid blocker, was approved by the Japanese health insurance system. Since its approval, patients refractory to vonoprazan can be encountered in clinical settings. We designed this study to clarify the pathophysiology of gastroesophageal reflux disease refractory to vonoprazan. Methods In this retrospective study, we involved patients who had refractory symptoms after administration of standard-dose proton pump inhibitors or vonoprazan and underwent diagnostic testing with esophageal high-resolution manometry and 24-hour multichannel intraluminal impedance and pH monitoring while using proton pump inhibitors or vonoprazan. Patients were diagnosed based on the Rome IV criteria for functional gastrointestinal disorders and diagnostic test results. Results Twenty-seven patients were analyzed during this study. Gastric pH ${\geq}4$ was sustained for a longer period of time, and the esophageal acid exposure time and number of acid reflux events were shorter in the vonoprazan group than in the proton pump inhibitor group. The percentage of patients diagnosed with acidic gastroesophageal reflux disease in the vonoprazan group was lower than that in the proton pump inhibitor group. Conclusions Intra-gastric pH and acid reflux were strongly suppressed by 20-mg vonoprazan. When patients with gastroesophageal reflux disease present symptoms after administration of 20-mg vonoprazan, the possibility of pathophysiologies other than acid reflux should be considered.
The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(-0.23~-0.92 and -0.68~-0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.
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