scholarly journals Increased Fertilization Rates after In Vitro Culture of Frozen-Thawed Testicular Immotile Sperm in Nonobstructive Azoospermic Patients

ISRN Urology ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
R. Nuñez-Calonge ◽  
S. Cortes ◽  
M. Gago ◽  
P. López ◽  
P. Caballero-Peregrin

Objective. To optimise the use of freeze/thaw testicular immotile spermatozoa from nonobstructive azoospermia patients and to analyse the outcome of intracytoplasmic sperm injection (ICSI) of such spermatozoa. Methods. Testicular specimens were retrieved and cryopreserved from forty patients with nonobstructive azoospermia and underwent one cycle with thawed spermatozoa (Group I) that led to pregnancy in sixteen cases. Twenty-four patients of group I underwent treatment with the same batch of thawed spermatozoa (Group II). For the first ICSI attempt, injection was performed when motile spermatozoa were found. In group II, injection was performed when maximum motility was reached. We compared mean of fertilization rate, embryo quality, clinical pregnancy rate and embryo implantation rate. Results. The mean percentage of motility was significantly higher in the group II than in the group I (18, 6 versus 8, 2). Group I showed a significant decrease in fertilization rates when compared with cryopreserved testicular spermatozoa in group II (54% versus 72%, ). No difference was noted between the cleavage rate, embryo quality, clinical pregnancy rates and implantation rates among group II and I. Conclusion. Fecundation rate can be significantly improved after in-vitro culture and sperm selection of frozen-thawed immotile testicular spermatozoa in patients with nonobstructive azoospermia.


2012 ◽  
Vol 24 (1) ◽  
pp. 138
Author(s):  
L. Boccia ◽  
M. Rubessa ◽  
M. De Blasi ◽  
S. Di Francesco ◽  
G. Albero ◽  
...  

Although in vitro embryo production efficiency in buffalos has greatly improved over the years, the in vitro-produced embryos show lower viability and resistance to cryopreservation. Therefore, it is necessary to optimize the in vitro culture conditions to improve embryo quality. Hyaluronic acid, a glycosaminoglican present in oviducal and uterine fluids, has been shown to successfully support in vitro development of bovine embryos (Stojkovic et al. 2002 Reproduction 124, 141–153). The aim of this study was to evaluate the influence of high concentrations of hyaluronic acid (HA) during late in vitro culture on blastocyst development, as well as on their cryotolerance after cryotop vitrification in buffalos. In vitro matured and fertilized buffalo oocytes (n = 1007) from slaughterhouse ovaries were cultured for 4 days in SOFaa supplemented by 8 mg mL–1 of BSA in a controlled gas atmosphere consisting of 5% CO2, 7% O2 and 88% N2, in humidified air, at 38.5°C. On Day 4, cleavage rate was assessed (75.2%) and all of the cleaved elements were divided into 3 different late culture groups: 8 mg mL–1 of BSA (n = 244; group A), 8 mg mL–1 of BSA supplemented by 6 mg mL–1 of HA (n = 251; group B) and 1 mg mL–1 of BSA supplemented by 6 mg mL–1 of HA (n = 262; group C). On Day 7 after IVF, embryo outcome was assessed and all of the embryos were vitrified by cryotop [De Rosa et al. 2007 Ital. J. Anim. Sci. 6 (Suppl 2), 747–750] and cultured for 24 h. The resistance to cryopreservation was evaluated by assessing the survival rate on the basis of morphological criteria and the percentage of embryos reaching a more advanced developmental stage after 24 h culture. Data were analysed by the chi-square test. No differences in blastocyst rate were recorded among groups (43.9, 44.3 and 40.0%, respectively in A, B and C groups). However, out of the total embryos, a higher percentage of Grade 1 hatched blastocysts (Robertson and Nelson 1998 Manual of the International Embryo Transfer Society 9, 103–16) was observed in group C (P < 0.05) than in groups A and B (14.3, 18.8 and 25.5% in A, B and C groups, respectively). Although the supplementation with HA did not improve the survival rates following vitrification-warming (51.1, 59.4 and 58.4% in A, B and C groups, respectively), the percentage of vitrified-warmed embryos that resumed development and reached a more advanced developmental stage after culture increased (P < 0.01) in group C (20.7, 27.7 and 37.6% in A, B and C groups, respectively). In conclusion, the addition of 6 mg mL–1 of HA, together with a limited protein source (i.e. 1 mg mL–1 of BSA), during late culture improved buffalo embryo quality, indicated by both the greater percentage of advanced-stage embryos and by the resumption of development after post-warming culture.



2013 ◽  
Vol 25 (5) ◽  
pp. 737 ◽  
Author(s):  
M. Castillo-Martín ◽  
M. Yeste ◽  
R. Morató ◽  
T. Mogas ◽  
S. Bonet

The objective of the present study was to determine the effects of replacing glucose with pyruvate and lactate during the first 48 h of in vitro culture (IVC) in NCSU-23 medium on embryo development, embryo quality and survival of porcine blastocysts after vitrification. To this end, in vitro-produced (IVP) porcine oocytes were cultured with either glucose for 6 days (IVC-Glu) or pyruvate–lactate from Day 0 to Day 2 and then with glucose until Day 6 (IVC-PyrLac). Blastocysts were vitrified on Day 6 using the Cryotop device and, after warming, survival rate and the apoptosis index were evaluated after 24 h incubation in NCSU-23 medium. No significant differences were observed between IVC-Glu and IVC-PyrLac in terms of cleavage rate, blastocyst yield, total number of cells per blastocyst or the apoptosis index (1.82 ± 0.75% vs 3.18 ± 0.88%, respectively) of non-vitrified embryos. However, a significant increase was seen in hatching/hatched blastocysts in the IVC-PyrLac compared with IVC-Glu treatment group (12.71 ± 1.20% vs 3.54 ± 0.47%, respectively). Regardless of treatment, vitrification impaired the survival rate and the apoptosis index. When comparing both treatments after warming, the percentage of apoptotic cells was significantly higher for blastocysts in the IVC-PyrLac compared with IVC-Glu group (18.55 ± 3.49% vs 9.12 ± 2.17%, respectively). In conclusion, under the conditions of the present study, replacement of glucose with pyruvate–lactate during the first 48 h of culture resulted in a lower cryotolerance of IVP porcine embryos.



Author(s):  
Minu Xaviour ◽  
Abhilash R.S. ◽  
Jayakumar C. ◽  
Amritha Aravind ◽  
Raji K.

Study evaluated the role of cAMP modulator (Forskolin and 3-isobutyl-1- methyl xanthine) supplementation on developmental competence of bovine oocytes. Cumulus oocyte complexes recovered from bovine ovaries of unknown reproductive status were used for the study. Oocytes retrieved by aspiration method were graded based on cumulus cell distribution and culture grade oocytes were selected for the study. A total of 414 culture grade oocytes were taken and divided into two groups. Group I constituted of 201 oocytes in which pre-maturation was carried out for a period of 2 h. In group II, 213 oocytes were selected in which normal maturation was carried out. Maturation was assessed after 24h of culture in CO2 incubator maintained at 38.5o C in 95 per cent humidified atmosphere of 5 per cent CO2 . Fertilisation was carried out using frozen thawed semen and the presumptive zygotes were then transferred to culture media and cleavage was assessed 48 h after insemination. A significantly higher maturation rate (p ?0.05) was observed in group I compared to group II (86.85 ± 1.19 vs 79.88 ± 2.67). There was a highly significant increase (p ?0.01) in cleavage rate in group I (65.92 ± 1.23) compared to group II (59.29 ± 1.50). A higher fertilisation rate was observed in group I (75.35 ± 1.19) than group II (71.88 ± 2.56). It could be concluded that pre-maturation with cAMP modulators improved the developmental competence of bovine oocytes.



1997 ◽  
Vol 36 (08) ◽  
pp. 259-264
Author(s):  
N. Topuzović

Summary Aim: The purpose of this study was to investigate the changes in blood activity during rest, exercise and recovery, and to assess its influence on left ventricular (LV) volume determination using the count-based method requiring blood sampling. Methods: Forty-four patients underwent rest-stress radionuclide ventriculography; Tc-99m-human serum albumin was used in 13 patients (Group I), red blood cells was labeled using Tc-99m in 17 patients (Group II) in vivo, and in 14 patients (Group III) by modified in vivo/in vitro method. LV volumes were determined by a count-based method using corrected count rate in blood samples obtained during rest, peak exercise and after recovery. Results: In group I at stress, the blood activity decreased by 12.6 ± 5.4%, p <0.05, as compared to the rest level, and increased by 25.1 ± 6.4%, p <0.001, and 12.8 ± 4.5%, p <0.05, above the resting level in group II and III, respectively. This had profound effects on LV volume determinations if only one rest blood aliquot was used: during exercise, the LV volumes significantly decreased by 22.1 ± 9.6%, p <0.05, in group I, whereas in groups II and III it was significantly overestimated by 32.1 ± 10.3%, p <0.001, and 10.7 ± 6.4%, p <0.05, respectively. The changes in blood activity between stress and recovery were not significantly different for any of the groups. Conclusion: The use of only a single blood sample as volume aliquot at rest in rest-stress studies leads to erroneous estimation of cardiac volumes due to significant changes in blood radioactivity during exercise and recovery.



Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
H. Debbarh ◽  
N. Louanjli ◽  
S. Aboulmaouahib ◽  
M. Jamil ◽  
L. Ahbbas ◽  
...  

Summary Maternal age is a significant factor influencing in vitro fertilization (IVF) outcomes. Oxidative stress (OS) is one of the major causes of age-related cellular and molecular damage. The purpose of this work was to investigate the correlation between maternal age with intrafollicular antioxidants and OS markers in follicular fluid (FF), and also to determine the OS status in patients of advanced age. This study was a prospective study including 201 women undergoing IVF whose age was between 24 and 45 years old. FF samples were obtained from mature follicles at the time of oocyte retrieval. After treatment of FF, lipid peroxidation levels (MDA) and enzyme activities such as superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione (GSH) level were evaluated using spectrophotometry. The results indicated that the age cutoff point for increasing the MDA level was fixed at 37 years, allowing the study to be differentiated into two age groups. Group I included patients whose age was less than 37 years, and group II included patients whose age was greater than or equal 37 years. Statistical analysis revealed that MDA and GSH levels and GR activity were significantly higher in group II compared with group I. The SOD and CAT activities were significantly less in group II compared with group I. We concluded that from 37 years old a reproductive ageing was accompanied by a change in the antioxidant pattern in FF that impaired reactive oxygen species scavenging efficiency.



2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 172-173
Author(s):  
Xiaoge Sun ◽  
Yitong Su ◽  
YangYi Hao ◽  
Gaokun Liu ◽  
Yue Gong ◽  
...  

Abstract To improve the efficiency of whole cottonseed (WCS) on dairy cattle, crush and alkali treatment were combined (Table 1) in this study. The gastrointestinal tract digestibility of WCS were measured by in situ ruminal incubation and in vitro fermentation, respectively. Finally, animal feeding experiment was conducted to evaluate the effect of processed WCS on dairy cattle. A total of 30 Holstein dairy cows with similar physiological status were allocated to three groups (CON, Group I, and Group II). Ration for the CON group was a basic ration with no WCS, ration with about 8% non-processed WCS was for Group I (DM basis), while the Group II was provided by the ration added with 8% Crush-Alkali (4% mixed alkali solids with NaOH/CaO ratio at 1:1) treated WCS. Results of the in situ and in vitro experiments showed that, compared with non-treated WCS, Crushing-Alkali treatment groups significantly improved WCS ruminal effective degradation rate of dry matter (DM), crude protein (CP), ether extract (EE), and neutral detergent fiber (NDF) (P &lt; 0.05). Furthermore, Crush-Alkali (4% mixed alkali solids with NaOH/CaO ratio at 1:1) was the most beneficial to improve WCS gastrointestinal degradation of DM, CP and NDF, as well as enhance WCS rumen-bypass of EE. With the lowest free gossypol content (P&lt; 0.05). Results of animal feeding experiment showed that DMI, 4% fat corrected milk production (4% FCM), milk protein, milk fat, and content of short-chain saturated fatty acids in milk in Group II significantly increased (P &lt; 0.05) compared with CON group. DMI, ω-6 polyunsaturated fatty acids (PUFA) in milk, and the ratio of ω-6 to ω-3 PUFA were significantly higher in Group II than that in Group I (P &lt; 0.05). Additionally, free gossypol concentration in serum and milk, as well as parameters reflecting liver function were not notably different among the three groups (P &gt; 0.05).



2012 ◽  
Vol 17 (4) ◽  
pp. 1-7 ◽  
Author(s):  
Roberto Hideo Shimizu ◽  
Karlos Giovani Grando ◽  
Isabela Almeida Shimizu ◽  
Augusto Ricardo Andriguetto ◽  
Ana Cláudia Moreira Melo ◽  
...  

OBJECTIVE: This in vitro study was designed to evaluate the shear bond strength (SBS) of orthodontic metal brackets bonded by direct and indirect techniques. METHODS: Thirty healthy human maxillary premolar teeth were used. The teeth were divided into three groups of 10 teeth each: Group I - indirect bonding with SondhiTM Rapid-Set system (3M/Unitek), Group II - indirect bonding with TransbondTM XT adhesive system (3M/Unitek) and Group III - direct bonding with TransbondTM XT adhesive system (3M/Unitek). After bonding and obtaining the specimens for the study, the specimens were subjected to SBS testing in a universal testing machine (Emic, model DL-500). The Kolmogorov-Smirnov test was applied to ascertain that the data had a normal distribution and the Bartlett test to check whether there was homogeneity of variance. One-factor analysis of variance was performed and, subsequently, Tukey's test for paired means. A 5% significance level was adopted. RESULTS: The results of Group I were 67.6 (N) and 5.9 (MPa); Group II, 68.9 (N) and 6.1 (MPa) and Group III (control), 92.5 (N) and 8.1 (MPa). CONCLUSION: It can therefore be concluded that the means for Group III were significantly higher compared with Groups I and II in both Newton (N) and Megapascal (MPa) values. The means attained by the indirect bonding technique used in Groups I and II, however, exhibited no statistically significant differences.



2009 ◽  
Vol 66 (1) ◽  
pp. 57-62 ◽  
Author(s):  
Drenka Turjacanin-Pantelic ◽  
Dragana Bojovic-Jovic ◽  
Biljana Arsic ◽  
Eliana Garalejic

Background/Aim. A modern approach to surgical treatment of tuboperitoneal infertility is based on laporascopic techniques. The aim of this study was to compare results of tuboperitoneal infertility treatment by the use of laparoscopy and classical laparotomy. Methods. A retrospectiveprospective study on 66 women treated operatively form tuboperitoneal infertility was performed. Data from patient's anamnesis and those related to the surgical treatment results, obtained by the use of an inquiry, were used in retrospective and prospective analysis, respectively. Chi-square test was used in statistical analysis. P value < 0.05 was considered significant. Results. Classical laparotomy was used on 34 women in a period from 1996 to 1997, while 32 women were operated laparoscopically in a period from 1999 to 2000. The results were as follows: a total number of conceived women was 16 (24%), seven in the group I (20.6%) and nine in the group II (28.1%); 13 women were with one pregnancy, six in the group I (17.6%) and seven in the group II (22%). Twice pregnant were three women, one in the group I (2.9%) and two in the group II (6.2%). The resulting pregnancies were: five women with abortion spontaneous, two in the group I (5.9%) and three in the group II (9.4%); two women with extrauterine pregnancy in the group I (5.9%); three with pretemporal birth, one in the group I (2.9%) and two in the group II (6.2%), while six women were with the temporal birth, two in the group I (5.9%) and four in the group II (12.5%). Statistical analysis showed that there was no significant difference in the results between these two groups. Conclusion. Surgical treatment of tubeperitoneal infertility, regardless of the used methods (classical laparotomy or laparoscopy) was successful in a great number of women. These methods have a great advantage over in vitro fertilization, and they should not be ignored.



Author(s):  
Nandita Bansal

Introduction: Intracoronal bleaching is the simplest, least invasive, and least expensive means available to lighten non-vital discolored teeth. The objective of the study: To evaluate the intra coronal bleaching efficacy of 35% Carbamide peroxide relative to 35% Hydrogen peroxide and Sodium perborate using artificially stained extracted human teeth. Materials and Methods: Eighty extracted human permanent maxillary and mandibular anterior teeth and single-rooted premolars were artificially stained using whole human blood and root canal treatment was completed. They were bleached twice 7 days apart using: Group I (n=20) – 35% Carbamide peroxide gel, Group II (n=20) – 35% Hydrogen peroxide gel, Group III (n=20) – 2 gm Sodium perborate mixed with per ml of distilled water, Group IV (n=20)– Distilled water only (control). Evaluation of shade was performed on day 0,7,14. Results: On day 7, In Groups I and Group II, there was a reduction in the shade by 2 vita tab positions, whereas Group IIIlightened by 1-tab position only. On day 14, CP and HP lightened by overall 3-tab positions and SP by 2-tab positions. There were no statistical differences between the groups on day 7 and day 14. Conclusion: 35% Carbamide peroxide, 35% Hydrogen peroxide and Sodiumperborate were equally effective for Intra coronal bleaching.



2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
George Queiroz Vaz ◽  
Alessandra Viviane Evangelista ◽  
Cassio Alessandro Paganoti Sartorio ◽  
Maria Cecilia Almeida Cardoso ◽  
Maria Cecilia Erthal ◽  
...  

Background.The use of donated oocytes for in vitro fertilization treatment in patients with ovarian failure is universally recognized. But would patients with polycystic ovarian syndrome (PCOS) be a good choice for egg donation programs?Objective.Comparing the pregnancy rates of egg receptors from donor patients diagnosed with PCOS to receptors from donors without PCOS.Design.Retrospective cohort study.Methods.A total of 234 patients who had undergone egg reception program were separated into two groups: Group I, receptors from PCOS donors (n=36); Group II, receptors from donors without PCOS (n=198). Medical records were reviewed and the fertilization, implantation, and pregnancy rates were calculated.Results.PCOS patients had an average of 3.23 more oocytes retrieved, but there were no differences in the number of mature oocytes that were used for donation between the groups. We also observed that the number of transferred embryos was also not significantly different, as well as the fertilization and implantation rates. The clinical pregnancy rates were not significantly different: 28% and 26% in Group I and Group II, respectively.Conclusions.Women with PCOS should not be excluded from egg donation programs.



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