76 EFFECTS OF REDUCED ENVIRONMENTAL LIGHT ON THE IN VITRO MATURATION OF PIG OOCYTES

2016 ◽  
Vol 28 (2) ◽  
pp. 167
Author(s):  
L. Y. Parra-Forero ◽  
A. Góngora ◽  
S. Romo-García ◽  
E. P. López Damian ◽  
G. D. Mendoza ◽  
...  
Keyword(s):  
Light Exposure ◽  
Red Light ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Penicillin G ◽  
Ambient Light ◽  
Embryo Viability ◽  
Group 2 ◽  
Group 1

The effects of light on the steps of embryo manipulation have been described in several species, including humans. There are reports in which exposure of these cells to UV rays from sunlight affects them epigenetically, which could lead to meiotic arrest that would prevent normal maturation from the germinal vesicle (GV) to metaphase II (MII) stage, which would compromise subsequent embryo viability. Development to the blastocyst was not evaluated. The objective of the study was to observe the difference in maturation capacity (GV to MII) of prepubertal gilt oocytes under conditions of reduced ambient light. Thirty Duroc ovaries were recovered at slaughter and immersed in saline (0.9% NaCl) supplemented with penicillin-G (100 IU mL–1) and streptomycin sulfate (100 mg mL–1) at 29 to 34°C for transport to the laboratory where they were punctured with an 18-gauge needle attached to a 10-mL syringe. GV oocytes were selected with at least 3 layers of compact cumulus cells and were incubated with TCM-199 for 42 h in total with replacement with fresh maturation medium at 22 h. Oocytes were subsequently denuded using 0.1% hyaluronidase, fixed with 2% formaldehyde, and stained with Hoechst 33342 (10 min). Thereafter, oocytes were evaluated under a fluorescence microscope for the stage of meiosis: GV, metaphase I (MI), anaphase I/telophase I (ATI), and MII. GV oocytes were divided into two groups of 60 each as follows: Group 1 = handling in ambient light (1 change of culture medium and evaluation at the stereoscope); Group 2 = handling with reduced light (the change of medium was carried out in a dark room with red light on the stereoscope. Chi-square and Student’s t-test with Minitab 16.0 statistical program were used, and differences were considered significant at P < 0.05. There were significant differences in the percentages of maturation stages between Group 1 and Group 2, respectively: GV = 3.33, 3.33% (P = 0.45); MI = 20, 13.3% (P = 0.037); ATI = 33.3, 26.6% (P = 0.03); and MII = 43.3, 56.6% (P = 0.019). In conclusion, rate of maturation to MII was significantly higher with decreased light exposure. There was no difference in the GV groups probably because there was no manipulation at this stage. Further studies are necessary to determine the amount of light needed to optimize oocyte viability and to assess any potential effects on blastocyst production.

scholarly journals 78 SURVIVAL OF PORCINE OOCYTES AT GERMINAL VESICLE STAGE AFTER VITRIFICATION WITH OPEN PULLED STRAW METHOD

2005 ◽  
Vol 17 (2) ◽  
pp. 189 ◽  
Author(s):  
A. Bali Papp ◽  
T. Somfai ◽  
E. Varga ◽  
M. Marosán
Keyword(s):  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Penicillin G ◽  
National Committee ◽  
Ministry Of Education ◽  
Germinal Vesicle Breakdown ◽  
Maturation Medium ◽  
Maturation Rate ◽  
Nuclear Stage

The present study was performed to assess the survival of immature denuded or cumulus-covered porcine oocytes (COCs). Immature porcine oocytes were collected from 2–6 mm follicles of slaughterhouse ovaries and subjected to open pulled straw (OPS) vitrification, according to the method of Vajta et al. (1998 Mol Reprod. Dev. 51, 53–58). After vitrification, oocytes were matured in vitro for 48 h at 39°C, 5% CO2 in air. The maturation medium was TCM199 supplemented with 10% pig follicular fluid, 1.25 mM L-glutamine, 0.9 mM Na pyruvate, 150 μM cysteamine, 0.1 mg/mL streptomycin sulfate, 100 IU/mL PG penicillin g potassium, 10 IU/mL PMSG, and 25 IU/mL hCG. After IVM, to assess nuclear stage, all oocytes were fixed with acetic acid–alcohol (1:3) for at least three days and then stained with 0.1% orcein and examined under a phase-contrast microscope at 100× magnification. All data were analyzed by χ2 test (P < 0.05). Immediately after collection, all oocytes were at the germinal vesicle (GV) stage with an intact GV membrane. After vitrification, significantly fewer oocytes had normal morphology (intact plasma membrane) in the denuded and COC groups (4.7% and 8.5%, respectively) than did the denuded and COC control groups (95% and 92%, respectively). By the end of IVM, significantly fewer oocytes were surrounded by expanded cumulus after vitrification of COCs than were the COC controls (28.1% and 63.5%, respectively). After IVM, more of the COC control oocytes underwent germinal vesicle breakdown than did the denuded controls (95% and 78.2%, respectively); the rate of MII oocytes was higher for the COC controls than for the denuded controls (80% and 54.5%, respectively). After vitrification, the number of oocytes that underwent GVBD was significantly less for both the denuded and the COC groups (2.0% and 7.0%, respectively); the percentage of oocytes that reached MII was also lower (0.64% and 2.78%, respectively). Most of the vitrified oocytes had a damaged GV with disrupted membrane and cluster-like or scattered chromatin in both the denuded and the COC groups (96.4% and 90.7%, respectively). These data suggest that vitrification of cumulus-enclosed immature porcine oocytes is preferable compared to vitrification of denuded ones. Loss of cumulus cells compromises competence of oocytes to resume meiosis, which might result in a lower maturation rate after IVM. This research was supported by the grants of the Hungarian Scientific Research Fund (T 031758), the Hungarian National Committee of the Technical Development at the Ministry of Education (00796/2003), and the Ministry of Education (OM-KMUFA; BIO-00086/2002).

scholarly journals Effect of Cumulus cell co-culture and Protein Supplement on Success of in vitro Fertilization and Development of Pre-implanted Embryos in mice

2005 ◽  
Vol 10 ◽  
pp. 31
Author(s):  
Muhammad-Baqir M-R. Fakhrildin
Keyword(s):  
In Vitro Fertilization ◽  
Embryonic Development ◽  
Amniotic Fluid ◽  
Cumulus Cells ◽  
Protein Source ◽  
Vitro Fertilization ◽  
Group 2 ◽  
Normal Embryonic Development ◽  
Group 1

Successful oocyte fertilization and normal embryonic development of mice were considered the most important diagnostic criteria for the safety of materials and tools used for human in vitro fertilization and embryo transfer (IVF-ET). Therefore, we studied the influence of cumulus cells co-culture and protein supplement within culture medium on percentages of in vitro fertilization (IVF) and normal development of early stages of mouse embryo later. Oocytes were collected and treated with hyaluronidase to remove cumulus cells. Oocytes were divided into four groups namely: Group-1: Oocytes incubated within modified Earl’s medium (MEM) supplied with 10% inactivated bovine amniotic fluid as a protein source and cumulus cells; Group-2: Oocytes incubated with MEM supplied with cumulus cells only; Group-3: Oocytes incubated with MEM supplied with 10% inactivated bovine amniotic fluid only; and Group-4: Oocytes  incubated with MEM free of both protein source and cumulus cells. For IVF, 5-6 oocytes were incubated with active spermatozoa under paraffin oil for 18-20 hours at 37° oC in 5% CO2. Percentages of IVF and embryonic development were then recorded. Best results for IVF and normal embryonic development were achieved from oocytes of Group-1 when compared to the other groups. As compared to Group-1, the percentage of IVF for Group-2 and Group-3 were decreased insignificantly and significantly (P<0.002), respectively. Significant (P<0.01) reduction in the percentages of IVF and normal embryonic development were reported in Group-4 as compared to Group-1. Therefore, it was concluded that the presence of cumulus cells co-culture and bovine amniotic fluid as a protein source within culture medium may have an important role on the fertilizing capacity of spermatozoa and oocytes and normal development of pre-implanted mouse embryo later.  

308 EFFECT OF GONADOTROPIN TREATMENT ON OOCYTE COLLECTION AND IN VITRO FERTILIZATION IN THE BAN MINIPIG

2007 ◽  
Vol 19 (1) ◽  
pp. 269 ◽  
Author(s):  
B. X. Nguyen ◽  
T. Nagai ◽  
K. Kukuchi ◽  
N. T. Uoc ◽  
M. Ozawa ◽  
...  
Keyword(s):  
Cumulus Cells ◽  
Treated Group ◽  
Vitro Fertilization ◽  
Oocyte Collection ◽  
First Results ◽  
Morula Stage ◽  
Serum Gonadotropin ◽  
Group 2 ◽  
Group 1

The Ban minipig is a local breed characterized by small ovaries with a scant number of follicles available for in vitro maturation (IVM). The combination of eCG and hCG has been used successfully to control estrus in pig breeding programs. In this paper we present the first results of IVF in this breed in comparison with 2 types of oocyte preparation: (1) from animals not receiving gonatotropin treatment (group 1, n = 9); and (2) from animals receiving an injection of mixed pregnant mare serum gonadotropin and hCG, 300 IU/animal, for 3 days before oocyte collection (group 2, n = 4). All animals were 1 to 3 years old and with body weights that varied from 8 to 12 kg. At the time of collection, the ovaries were observed for follicle development; the cumulus–oocyte complexes (COCs) were aspirated using a 18-gauge needle. COCs of categories A (with more than 4 layers of cumulus cells) and B (with 2 to 4 layers of cumulus cells) were collected and matured in vitro as described previously (Kikuchi et al. 2002 Biol. Reprod. 66, 1033–1041) at 39�C under 5% CO2 in air. Matured oocytes with expanding cumulus cells were inseminated using male Ban minipig epididymal semen frozen by the methods reported by Kikuchi et al. (1998 Theriogenology 50, 615–623). The frozen–thawed spermatozoa were pre-incubated for 1 h in modified medium-199 adjusted to pH 7.8 in the incubator at 37�C. The capacitated spermatozoa were diluted and added to drops of fertilization medium (Fig-FM; Suzuki et al. 2002 Int. J. Androl. 123, 135–142) containing oocytes; the final concentration of sperm was 106/mL. After 3 h of co-incubation, attached spermatozoa and cumulus cells were removed from oocytes and the oocytes were the cultured in vitro as described previously (Kikuchi et al. 2002). The results obtained from 4 replicates showed that the number of follicles with a diameter larger than 2 mm and the rates of oocytes categorized as A and B were significantly lower (P &lt; 0.05; ANOVA test) in the nontreated animals (0.0 and 67.5%, respectively) than in the treated group (25.5 and 87.1%, respectively). The rates of oocytes with a clearly expanding cumulus obtained after IVM were 78.6 (n = 136) and 88.1% (n = 101) for groups 1 and 2, respectively. The rates of cleaved embryos and embryos developed to the compact morula stage were 47.2 and 9.1% (n = 39), respectively, for group 1; and 89.1 and 18.8% (n = 101), respectively, for group 2. In conclusion, gonadotropin treatment before the collection of oocytes is recommended for application of IVM–IVF to local Ban minipigs. This work was supported by a grant from the VAST-Japan Society for the Promotion of Science Project.

266 THE SHORT TIME TREATMENT WITH SODIUM BUTYRATE ON GERMINAL VESICLE STAGE OOCYTES IMPROVES OOCYTE QUALITY AND DEVELOPMENTAL COMPETENCE IN PIGS

2011 ◽  
Vol 23 (1) ◽  
pp. 231
Author(s):  
L. M. Liu ◽  
F. Gao ◽  
M. Hua ◽  
J. Y. Guan ◽  
B. Tang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Sodium Butyrate ◽  
Germinal Vesicle ◽  
Developmental Competence ◽  
Control Group ◽  
Blastocyst Rate ◽  
Group 2 ◽  
Short Time ◽  
Group 1

Oocyte maturation is a complex process during which the epigenetic modifications are dramatically changed, especially the histone acetylation and phosphorylation. Sodium butyrate (NaBu) is a histone deacetylase inhibitor that results in a more open structure of DNA. The aim of the present study was to analyse the role of NaBu in the meiosis of porcine oocytes and the subsequent embryonic developmental competence. Cumulus–oocyte complexes (COC) were collected from ovaries obtained at a local slaughterhouse. The COC were randomly divided into 3 groups and matured in vitro in medium (Hao et al. 2006) supplemented with 1 μM NaBu for 2 h [germinal vesicle (GV) stage, group 1] or for 22 h [GV to GV breakdown (GVBD) stage, group 2] or without treatment (control, group 3). After 44 h of in vitro maturation, the oocytes were denuded by 0.2% hyaluronidase, and oocytes with evenly dark ooplasm and visible first polar bodies were considered matured. The cortical granule distribution of matured oocytes was examined with immunostaining. The relative expression of CyclinB and Cdc2 of 3 group oocytes was determined with real-time PCR. Some matured oocytes from each group were collected and stimulated with electric pulse (2 direct current pulses of 1.2 kV cm–1 for 30 μs). The rate of parthenogenetic blastocyst was recorded, and cell number of each blastocyst was determined under an inverted fluorescence microscope after staining with 10 μg mL–1 of Hoechst 33342. The following results were found. 1) Compared with the control group (n = 70, 67.74 ± 1.64), oocyte maturation rates of group 1 and group 2 decreased significantly along with the extended treatments (n = 70, 59.57 ± 5.29 and 46.99 ± 1.22, respectively; P < 0.05). 2) The long time (22 h) treatment with NaBu inhibited the developmental competence (blastocyst rate) of oocytes (n = 30, 15.33 ± 3.47 v. 27.16 ± 2.10 P < 0.05), and the short time (2 h) treatment with NaBu on GV-stage oocytes inhibited the meiotic process slightly but improved the blastocyst rate (n = 30, 33.93 ± 2.51 v. 27.16 ± 2.10; P < 0.05). 3) The short time (2 h) treatment resulted in the migration of more cortical granules into the plasmasmic membrane and formed a monolayer with the membrane (compared with the control). 4) The exposure to NaBu from GV to GVBD stage induced the expression of the CyclinB and Cdc2 in the matured oocytes (4.68 ± 0.45 and 5.80 ± 0.58, respectively; P < 0.05) compared with the control. 5) Short time (2 h) exposure to NaBu on GV-stage oocytes inhibited the expression of the Cdc2 but increased the expression of the CylinB in the matured oocytes (0.43 ± 0.06 and 1.65 ± 0.26, respectively; P < 0.05) compared with the control. In conclusion, results of this study demonstrate that exposure to NaBu inhibits porcine oocyte meiosis in proportion to treatment length. However, a 2-h treatment with 1 μM NaBu improves oocyte developmental competence to the blastocyst stage. These results are useful for improving the developmental competent of oocytes for IVF and in vitro embryo production. This work was supported by a grant (No. 2009CB941001) from the National Basic Research Program of China.

scholarly journals Cryopreservation of Immature Oocytes at Germinal Vesicle Stage. When Gamete Maturation Performance Seems to Be Most Appropriate?

2021 ◽  
Vol 31 (2) ◽  
pp. 161-167
Author(s):  
Taisiia Yurchuk ◽  
Keyword(s):  
Embryo Development ◽  
Ovarian Tissue ◽  
Reproductive Potential ◽  
Germinal Vesicle ◽  
Immature Oocytes ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Fertility preservation is among the priorities in reproductive medicine. However, the cancer patients and women with various functional ovarian disorders, wishing to preserve future reproductive potential may have some contraindications or no possibilities to cryopreserve mature oocytes and ovarian tissue. Therefore, the development of techniques for immature oocyte cryopreservation is considered an alternative strategy. Here, we have evaluated the survival, maturation, fertilization and embryo development rates of immature oocytes (Germinal vesicle (GV) stage – group 1) after cryopreservation and in vitro matured (IVM) ones (group 2) prior to cryopreservation, compared with in vivo matured metaphase-II (MII) oocytes (group 3). Survival rates were 97.6, 96.2 and 98.2 % for groups 1–3, respectively. The maturation rate of GV oocytes in group 1 was significantly lower than in group 2 and made 52.0 and 73.2%, respectively. The highest fertilization rate was revealed in group 3, and the lowest one was in group 1. The groups 1–3 showed the same tendency for further embryo development, i. e. the blastulation rates were 20.0, 38.5 and 56.9%, respectively. Thus, the survival rate of cryopreserved oocytes did not depend on their maturity rate. However, the IVM oocytes displayed lower fertilization and blastulation rates, than the in vivo matured ones. It was found that oocytes IVM should be performed prior to cryopreservation, because it ensured higher rates of maturation, fertilization and embryo development in vitro.

scholarly journals Effect of Splinting in Accuracy of Two Implant Impression Techniques

2014 ◽  
Vol 40 (6) ◽  
pp. 633-639 ◽  
Author(s):  
Erica Dorigatti de Avila ◽  
Fernanda de Matos Moraes ◽  
Sabrina Maria Castanharo ◽  
Marcelo Antonialli Del'Acqua ◽  
Francisco de Assis Mollo
Keyword(s):  
Confidence Interval ◽  
Analysis Of Variance ◽  
Impression Material ◽  
Titanium Screw ◽  
Standard Preparation ◽  
Implant Abutment ◽  
Interface Gaps ◽  
Group 2 ◽  
Group 1

Because there is no consensus in the literature about the need for a splint between copings, the aim of this study was to evaluate, in vitro, the accuracy of 2 impression techniques for implant-supported prostheses. A master cast was fabricated with four parallel implant abutment analogs and a passive framework. Two groups with 5 casts each were formed: Group 1 (squared impression copings with no splint: S) and Group 2 (splinted squared impression copings, using metal drill burs and Pattern resin: SS). The impression material used was polyvinyl siloxane with open trays for standard preparation of the casts. For each cast, the framework was positioned, and a titanium screw was tightened with 10 N·cm torque in analog A, after which measurements of the abutment-framework interface gaps were performed at analogs C and D. This process was repeated for analog D. These measurements were analyzed using software. A one-way analysis of variance (ANOVA) with a confidence interval of 95% was used to analyze the data. Significant differences were detected between S and SS in relation to the master cast (P ≤ 0.05). The median values of the abutment-framework interface gaps were as follows: master cast: 39.64 μm; squared impression copings with no splint: 205.86 μm; splinted squared impression copings: 99.19 μm. Under the limitations of this study, the technique presented for Group 2 produces better results compared with the technique used for Group 1.

scholarly journals A Flexible Multidose GnRH Antagonist versus a Microdose Flare-Up GnRH Agonist Combined with a Flexible Multidose GnRH Antagonist Protocol in Poor Responders to IVF

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Gayem İnayet Turgay Çelik ◽  
Havva Kömür Sütçü ◽  
Yaşam Kemal Akpak ◽  
Münire Erman Akar
Keyword(s):  
Gnrh Agonist ◽  
Gnrh Antagonist ◽  
Poor Responders ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Gnrh Antagonist Protocol ◽  
Group 2 ◽  
Antagonist Protocol ◽  
Group 1

Objective. To compare the effectiveness of a flexible multidose gonadotropin-releasing hormone (GnRH) antagonist against the effectiveness of a microdose flare-up GnRH agonist combined with a flexible multidose GnRH antagonist protocol in poor responders to in vitro fertilization (IVF).Study Design. A retrospective study in Akdeniz University, Faculty of Medicine, Department of Obstetrics and Gynecology, IVF Center, for 131 poor responders in the intracytoplasmic sperm injection-embryo transfer (ICSI-ET) program between January 2006 and November 2012. The groups were compared to the patients’ characteristics, controlled ovarian stimulation (COH) results, and laboratory results.Results. Combination protocol was applied to 46 patients (group 1), and a single protocol was applied to 85 patients (group 2). In group 1, the duration of the treatment was longer and the dose of FSH was higher. The cycle cancellation rate was significantly higher in group 2 (26.1% versus 38.8%). A significant difference was not observed with respect to the number and quality of oocytes and embryos or to the number of embryos transferred. There were no statistically significant differences in the hCG positivity (9.5% versus 9.4%) or the clinical pregnancy rates (7.1% versus 10.6%).Conclusion. The combination protocol does not provide additional efficacy.

scholarly journals THU0227 CAFFEINE INTAKE MODULATES DISEASE ACTIVITY AND CYTOKINES LEVELS IN SYSTEMIC LUPUS ERYTHEMATOSUS PATIENTS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 340.2-341
Author(s):  
V. Orefice ◽  
F. Ceccarelli ◽  
C. Barbati ◽  
R. Lucchetti ◽  
G. Olivieri ◽  
...  
Keyword(s):  
Disease Activity ◽  
Lupus Erythematosus ◽  
Caffeine Intake ◽  
Caffeine Consumption ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
The Impact ◽  
Group 1

Background:Systemic lupus erythematosus (SLE) is an autoimmune disease mainly affecting women of childbearing age. The interplay between genetic and environmental factors may contribute to disease pathogenesis1. At today, no robust data are available about the possible contribute of diet in SLE. Caffeine, one of the most widely consumed products in the world, seems to interact with multiple components of the immune system by acting as a non-specific phosphodiesterase inhibitor2.In vitrodose-dependent treatment with caffeine seems to down-regulate mRNA levels of key inflammation-related genes and similarly reduce levels of different pro-inflammatory cytokines3.Objectives:We evaluated the impact of caffeine consumption on SLE-related disease phenotype and activity, in terms of clinimetric assessment and cytokines levels.Methods:We performed a cross-sectional study, enrolling consecutive patients and reporting their clinical and laboratory data. Disease activity was assessed by SLE Disease Activity Index 2000 (SLEDAI-2k)4. Caffeine intake was evaluated by a 7-day food frequency questionnaire, including all the main sources of caffeine. As previously reported, patients were divided in four groups according to the daily caffeine intake: <29.1 mg/day (group 1), 29.2-153.7 mg/day (group 2), 153.8-376.5 mg/day (group 3) and >376.6 mg/day (group 4)5. At the end of questionnaire filling, blood samples were collected from each patient to assess cytokines levels. These were assessed by using a panel by Bio-Plex assays to measure the levels of IL-6, IL-10, IL-17, IL-27, IFN-γ, IFN-α and Blys.Results:We enrolled 89 SLE patients (F/M 87/2, median age 46 years, IQR 14; median disease duration 144 months, IQR 150). The median intake of caffeine was 195 mg/day (IQR 160.5). At the time of the enrollment, 8 patients (8.9%) referred a caffeine intake < 29.1 mg/day (group 1), 27 patients (30.3%) between 29.2 and 153.7 mg/day (group 2), 45 patients (51%) between 153.8 and 376.5 mg/day (group 3) and 9 patients (10.1%) >376.6 mg/day (group 4). A negative correlation between the levels of caffeine and disease activity, evaluated with SLEDAI-2K, was observed (p=0.01, r=-0.26). By comparing the four groups, a significant higher prevalence of lupus nephritis, neuropsychiatric involvement, haematological manifestations, hypocomplementemia and anti-dsDNA positivity was observed in patients with less intake of caffeine (figure 1 A-E). Furthermore, patients with less intake of caffeine showed a significant more frequent use of glucocorticoids [group 4: 22.2%,versusgroup 1 (50.0%, p=0.0001), group 2 (55.5%, p=0.0001), group 3 (40.0%, p=0.009)]. Moving on cytokines analysis, a negative correlation between daily caffeine consumption and serum level of IFNγ was found (p=0.03, r=-0.2) (figure 2A); furthermore, patients with more caffeine intake showed significant lower levels of IFNα (p=0.02, figure 2B), IL-17 (p=0.01, figure 2C) and IL-6 (p=0.003, figure 2D).Conclusion:This is the first report demonstrating the impact of caffeine on SLE disease activity status, as demonstrated by the inverse correlation between its intake and both SLEDAI-2k values and cytokines levels. Moreover, in our cohort, patients with less caffeine consumption seems to have a more severe disease phenotype, especially in terms of renal and neuropsychiatric involvement. Our results seem to suggest a possible immunoregulatory dose-dependent effect of caffeine, through the modulation of serum cytokine levels, as already suggested byin vitroanalysis.References:[1]Kaul et alNat. Rev. Dis. Prim.2016; 2. Aronsen et alEurop Joul of Pharm2014; 3. Iris et alClin Immun.2018; 4. Gladman et al J Rheumatol. 2002; 5. Mikuls et alArth Rheum2002Disclosure of Interests:Valeria Orefice: None declared, Fulvia Ceccarelli: None declared, cristiana barbati: None declared, Ramona Lucchetti: None declared, Giulio Olivieri: None declared, enrica cipriano: None declared, Francesco Natalucci: None declared, Carlo Perricone: None declared, Francesca Romana Spinelli Grant/research support from: Pfizer, Consultant of: Novartis, Gilead, Lilly, Sanofi, Celgene, Speakers bureau: Lilly, cristiano alessandri Grant/research support from: Pfizer, Guido Valesini: None declared, Fabrizio Conti Speakers bureau: BMS, Lilly, Abbvie, Pfizer, Sanofi

scholarly journals Evaluation of Gingival Microleakage in Class II Composite Restorations with Different Lining Techniques: An In Vitro Study

Scientifica ◽  
2015 ◽  
Vol 2015 ◽  
pp. 1-6
Author(s):  
Vedavathi Bore Gowda ◽  
B. V. Sreenivasa Murthy ◽  
Swaroop Hegde ◽  
Swapna Devarasanahalli Venkataramanaswamy ◽  
Veena Suresh Pai ◽  
...  
Keyword(s):  
Study Groups ◽  
Class Ii ◽  
Composite Liner ◽  
Flowable Composite ◽  
Composite Restorations ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Aim. To compare the microleakage in class II composite restorations without a liner/with resin modified glass ionomer and flowable composite liner.Method. Forty standardized MO cavities were prepared on human permanent mandibular molars extracted for periodontal reasons and then divided into 4 groups of ten specimens. The cavity preparations were etched, rinsed, blot dried, and light cured and Adper Single Bond 2 is applied. Group 1 is restored with Filtek P60 packable composite in 2 mm oblique increments. Group 2 is precure group where 1 mm Filtek Z350 flowable liner is applied and light cured for 20 sec. Group 3 is the same as Group 2, but the liner was cocured with packable composite. In Group 4, 1 mm RMGIC, Fuji Lining LC is applied and cured for 20 sec. All the teeth were restored as in Group 1. The specimens were coated with nail varnish leaving 1 mm around the restoration, subjected to thermocycling, basic fuchsin dye penetration, sectioned mesiodistally, and observed under a stereomicroscope.Results. The mean leakage scores of the individual study groups were Group 1 (33.40), Group 2 (7.85), Group 3 (16.40), and Group 4 (24.35). Group 1 without a liner showed maximum leakage. Flowable composite liner precured was the best.

scholarly journals Experimental Reproduction of Bovine Fetal Neospora Infection and Death with a Bovine Neospora Isolate

1994 ◽  
Vol 6 (2) ◽  
pp. 207-215 ◽  
Author(s):  
Bradd C. Barr ◽  
Joan D. Rowe ◽  
Karen W. Sverlow ◽  
Robert H. BonDurant ◽  
Alex A. Ardans ◽  
...  
Keyword(s):  
Uninfected Cell ◽  
In Utero ◽  
Pathogenic Potential ◽  
Group 4 ◽  
Fetal Infection ◽  
Group 3 ◽  
Group 2 ◽  
Protozoal Infection ◽  
Group 1

Studies were conducted to determine the pathogenic potential of the recently isolated bovine Neospora protozoa (BPA-1) for the bovine fetus. Cows chosen for study had Neospora titers < 160 using an indirect immunofluorescent antibody (IFA) test. Four experimental groups were studied. In group 1, 2 fetuses were inoculated in utero at 118 days gestation with culture-derived Neospora tachyzoites. A pregnant control cow was housed in the same pen, observed daily and screened serologically for evidence of exposure to Neospora. In group 2, 2 cows were infected with Neospora tachyzoites at 138 or 161 days gestation, and 1 control cow was given uninfected cell culture suspension simultaneously at 154 days gestation. Groups 3 (85 days gestation) and 4 (120 days gestation) each consisted of 2 cows infected with Neospora tachyzoites and 1 control cow given uninfected material at the same stage of gestation. Dead fetuses were surgically removed from the infected cows in group 1 on postinfection day (PID) 17. The histopathology was compatible with protozoal fetal infection, and protozoa were identified by immunohistochemistry. Viable fetuses were removed surgically from cows in group 2 on PID 28-30. The histopathology was compatible with protozoal fetal infection, protozoa were identified by immunoperoxidase techniques, and Neospora tachyzoites were reisolated in vitro from tissues of the 2 infected fetuses. In groups 3 and 4, the control fetus and 1 infected fetus were removed surgically between PID 26 and PID 33. The remaining infected cows were observed until fetal death or abortion occurred. In group 3, the fetus that was surgically removed from 1 infected cow had no pathologic abnormalities, and parasites were not found (PID 26). The second fetus in group 3 died in utero, and expulsion of a mummified fetus was induced on PID 67. Brain histopathology was compatible with protozoal infection, and parasites were identified by immunoperoxidase techniques. The fetus that was surgically removed (PID 32) from 1 infected cow in group 4 had lesions compatible with protozoal infection, and Neospora tachyzoites were reisolated in vitro from fetal tissues. The second infected cow in group 4 produced a full-term live calf that had a precolostral Neospora titer of 20,480. Clinically, this calf had depressed conscious proprioception in all limbs. Very mild lesions were found in the central nervous system, but protozoa were not found in the tissues. The results demonstrate that the bovine Neospora protozoa can be transplacentally transmitted, resulting in fetal infection and death, and mimics the naturally occurring fetal disease.

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