284 FERTILIZING CAPACITY OF BUFFALO (BUBALUS BUBALIS) SPERM CO-CULTURED WITH OVIDUCT EPITHELIAL CELLS

2010 ◽  
Vol 22 (1) ◽  
pp. 299 ◽  
Author(s):  
E. Mariotti ◽  
S. Di Francesco ◽  
M. De Blasi ◽  
C. Siniscalchi ◽  
M. V. Suárez ◽  
...  

The overall in vitro embryo production efficiency in buffalo is hampered by the poor IVF efficiency. The aim of this work was to evaluate whether the fertilizing ability of buffalo sperm is improved by the presence of bovine oviductal cells (BOEC) during IVF. Because of limited availability of buffalo oocytes, this was assessed by heterologous IVF. Bovine oviducts were obtained at a local abattoir from cows that were in the preovulatory phase of a normal estrous cycle. BOEC recovered from 5 oviducts as previously described (Gualtieri and Talevi 2000 Biol. Reprod. 62, 1754-1762) were pooled and plated in 100 μL drops of TCM-199 + 10% FCS, 100 U mL-1 penicillin, 100 μg mL-1 streptomycin and 0.25 μg mL-1 amphotericin B under mineral oil. Medium was changed every 48 h up to Day 6, when cell confluence and cilia activity were optimal. On day of IVF the medium was removed from the drops and replaced with TALP supplemented with 0.2 mM penicillamine, 0.1 mM hypotaurine, and 0.01 mM heparin (IVF medium). Frozen-thawed sperm from an IVF-tested buffalo bull, treated by Percoll gradients, were used for all IVF groups (2 × 106 sperm mL-1). In vitro-matured bovine oocytes (n = 409), over 3 replicates, were distributed in 4 fertilization groups: (A) IVF medium alone (control); (B) BOEC monolayer + IVF medium; (C) sperm preincubated for 6 h in IVF medium; and (D) sperm preincubated for 6 h with BOEC + IVF medium. After 20 h of coincubation at 38.5°C and 5% CO2 in air, putative zygotes were denuded, washed, and cultured in SOF medium. Forty-eight hours after IVF, cleavage rate was evaluated, and cleaved and uncleaved oocytes were fixed in 60% methanol and stained with DAPI for nuclei examination under fluorescence microscope. Data were analyzed by chi-square test. Although cleavage rate was not different among groups (46.2, 55.8, 50.0, and 50.0% for A, B, C, and D, respectively), the monospermic penetration rate increased (P < 0.01) in group B (79.3%) compared with group A (69.6%), with intermediate values in groups C (75.2%) and D (76.0%). Interestingly, the percentage of advanced embryos (>4 cells) was higher (P < 0.01) in groups C and D (47.9 and 37.1%, respectively) than in group A (12.1%), whereas group B (21.0%) was only different from group C. We demonstrated that the fertilizing capacity of buffalo sperm, evaluated as oocyte penetration rate after heterologous IVF, is enhanced by the presence of BOEC. This suggests that IVF of buffalo oocytes on BOEC monolayer may improve the IVF efficiency in buffalo. The higher incidence of advanced embryos in both groups with preincubated sperm may be accounted for by an earlier accomplishment of capacitation, leading to anticipated oocyte penetration. However, because the penetration rate in these groups was not improved compared with the control, we hypothesize that sperm viability may have decreased and hence that shorter incubation times should be tested in further studies.

2021 ◽  
Vol 45 (5) ◽  
pp. 312-316
Author(s):  
Mishra Neha Sanjeev ◽  
Harsimran Kaur ◽  
Sandeep Singh Mayall ◽  
Rishika ◽  
Ramakrishna Yeluri

Objective: To evaluate the effectiveness of placing a resorbable collagen barrier in impeding the extrusion of obturation material in primary molars undergoing resorption. Study design: All the 94 canals in 47 mandibular molars were allocated to 2 groups- Group ‘A’- 47 canals with collagen barrier (Test group) and Group ‘B’- 47 canals without collagen barrier (Control group) based on randomization protocol. Pulpectomy was performed and obturation of both test and control canals were radiographically assessed. Pearson’s chi – square test was applied to analyze the results. The significance level was predetermined at p &lt; 0.05. Results: Among the test group, 93.6% of the canals showed no extrusion while, 6.4% showed visible extrusion of the material outside the apex. In the control group, 83% showed no extrusion whereas 17% of the canals showed visible extrusion outside the apex. But no significant difference was noted (p&gt;0.05). Conclusion: The placement of resorbable collagen barrier in the apical third of the canal prevented the extrusion of obturating material beyond the apex in resorbing primary molars.


2019 ◽  
Vol 3 (9) ◽  
pp. 283-289
Author(s):  
Shuchi Sukul ◽  
Goldy Rathee ◽  
Parimal Anand ◽  
Sakshi Kataria ◽  
Pratibha Taneja

OBJECTIVES: The present research was conducted to assess incidences of root microcracks caused by hand and rotary file system at different lengths MATERIAL AND METHODS: This in-vitro study was undertaken to assess incidence of root microcracks caused by hand and rotary file system at different lengths In total, 100 the mandibular premolar with straight roots determined with intact, fully formed apices were taken. Samples were randomly distributed into 5 groups based on the file system used: a) Group A: Control, b) Group B: Reciproc, c) Group C: WaveOne, d) Group D: One Shape and e) Group E: ProTaper. Pearson Chi-square test was used to determine the differences between groups. The dentinal defects were expressed as percentage of samples with microcracks in each group. Level of statistical significance was set at p-value less than 0.05. RESULTS: The chi square test was used to compare the Distribution of the number of teeth in which cracks were observed on the horizontal sections. It was found to be significant with group ProTaper showing maximum cracks at 3,6,9 mm level as compared to other file systemCONCLUSION: Nickel-titanium instruments causes cracks on the apical root surface or in the root canal wall. ProTaper causes maximum dentinal cracks as compared to other file systems


2015 ◽  
Vol 87 (8) ◽  
Author(s):  
Santosh Kumar ◽  
Rajesh Roat ◽  
Swati Agrawal ◽  
Kumar Jayant ◽  
Ravimohan S. Mavuduru ◽  
...  

Abstractwas to assess efficacy of Tadalafil alone versus Tadalafil plus Pentoxifylline in the treatment of erectile dysfunction by using self administered IIEF-5 questionnaire.Two hundred and thirty seven patients presenting with ED at andrology OPD were evaluated for ED by a self administered IIEF (International Index of Erectile Function) questionnaire. Patients were systematically randomized by computer generated random table into two groups groups namely, group A: Tadalafil only group, group B: combination of Tadalafil + Pentoxifyl-line. All the patients were re-assessed by IIEF-5 questionnaire after 8 weeks of medical therapy. Statistical analysis was performed using student’s unpaired t-test, paired t-test, chi square test. p-value < 0.05 was considered statistically significant.Two hundred and thirty seven patients were included in the present study, in group A: 92 patients (78.6%) showed improvement in their IIEF score after 8 weeks of tadalafil treatment. While in group B, overall 104 patients(86.6%) showed improvement after combination of Tadalafil and Pentoxifylline. There was a statistically significant difference of percentage change in IIEF score was seen in group B (group A 90.7±15.2%, group B 95.6±13.4%; p value – 0.014). We found this difference even more statistically significant in patients with severe ED (group A 72.7±47.2%, group B 132.3±54.3%; p value – 0.000). There was no significant difference in between the two groups with regards to occur-rence of side effects.Both tadalafil and combination of Tadalafil + Pentoxifylline improve erectile function in patients of ED. Patients with severe ED showed much significant improvement in erectile function with combination therapy.


2006 ◽  
Vol 18 (2) ◽  
pp. 143
Author(s):  
D. Salamone ◽  
M. Catala ◽  
A. Gibbons ◽  
F. Pereyra Bonnet ◽  
M. Cueto

Different types of somatic cells have been used as nucleus donors for cloning. Most of them were previously cultured in vitro as a monolayer through several plate passages. The experiment reported here was conducted to study the potential usages of granulosa and cumulus cells for cloning without previous culture as a monolayer. A first-plate-passage fibroblast was also used. Oocytes were aspirated by laparoscopy from Criolla goats and matured in TCM-199 + 5% FCS at 39°C for 24 h. Matured oocytes were denuded by vortexing for 3 min in TL HEPES with 1 mg/mL bovine testis hyaluronidase. Metaphases were assessed and oocytes were enucleated by visualization with Hoechst 33342 (5 μg/mL) under UV light (<6 s). Granulosa and cumulus cells were also recovered by laparoscopy and maintained in maturation medium in cryotube for 20 h at room temperature or 39°C, respectively. Goat adult ear fibroblasts were cultured for 1 or 2 weeks and used 2 days after confluence. All types of donor cells were transferred to the perivitlline space of enucleated oocytes and fused by an electrical pulse. After 2 h, activation was induced by incubation in TL-HEPES with 5 µM ionomycin for 4 min and 2 mM 6-DMAP for 3 h. The oocytes were then washed with TL-HEPES and cultured in SOF medium and atmosphere of 5% CO2 + 5% O2 + 90% N2. Cleavage (Day 2) and development to blastocysts (Day 6) were recorded and analyzed by chi-square test. The cleavage rate for non-plated granulosa cells was higher than for the other treatment goups; cumulus cells had a lower rate of development to blastocysts (Table 1). These results suggest that granulosa cells collected and maintained for 24 h at room temperature could be used to produce cloned blastocysts. Table 1. Effect of non-plated granulosa and cumulus cells and first passage fibroblasts as donor nucleus oocytes in goat cloning


2009 ◽  
Vol 21 (1) ◽  
pp. 131 ◽  
Author(s):  
M. De Blasi ◽  
E. Mariotti ◽  
M. Rubessa ◽  
S. Di Francesco ◽  
G. Campanile ◽  
...  

Despite the increasing interest, buffalo oocyte cryopreservation is still inefficient, especially in terms of blastocyst development after IVF. The aim of this work was to evaluate chromatin and spindle organization of buffalo in vitro-matured oocytes after vitrification/warming by cryotop and after their simple exposure to cryoprotectants (CP). An overall amount of 251 COC was selected and matured in vitro. In the vitrification group, COC were first exposed to 10% ethylene glycol (EG) + 10% DMSO for 3 min, and then to 20% EG + 20% of DMSO and 0.5 m sucrose, loaded on cryotops, and plunged into liquid nitrogen within 25 s. Oocytes were warmed into a 1.25 m sucrose solution for 1 min and then to decreasing concentrations of sucrose (0.625 m, 0.42 m, and 0.31 m) for 30s each. In order to test CP toxicity, COC were simply exposed to the vitrification and warming solutions. Two hours after warming, oocytes were fixed and immunostained for microtubules using a method previously described (Messinger SM and Albertini DF 1991 J. Cell Sci. 100, 289–298), stained for nuclei with Hoechst, and examined by fluorescence microscopy. Fresh in vitro-matured oocytes were fixed and stained as controls. Data were analyzed by chi-square test; results are shown in Table 1. The percentages of MII oocytes in the control and vitrification groups were greater than in the toxicity group, in which a greater percentage of telophase II stage oocytes were found compared with both the control and vitrification groups, indicating occurrence of activation. Of the MII oocytes, both exposure to CP and vitrification procedures gave greater percentages of oocytes with abnormal spindle and abnormal chromatin configuration compared with the control. An unexpected datum was the evidence of a significant percentage of spontaneously activated oocytes in the toxicity group. We speculate that the lack of activation in the vitrification group may be related to the slowing down of metabolic activity subsequent to thermal shock, and hence, that activation after vitrification may occur later than 2 h post-warming. In conclusion, the simple exposure to CP causes activation of the COC and damage to the cytoskeleton similar to that induced by the whole vitrification protocol. The damages to the meiotic spindle and DNA fragmentation may lead to aneuploidy incompatible with subsequent embryo development and account for the poor embryo development currently recorded in buffalo. Table 1.Chromatin and spindle organization in oocytes vitrified and exposed to cryoprotectants


2010 ◽  
Vol 22 (1) ◽  
pp. 317
Author(s):  
I. Venditto ◽  
E. Mariotti ◽  
L. Boccia ◽  
M. Rubessa ◽  
M. De Blasi ◽  
...  

Fertilization is a critical step of the in vitro embryo production (IVEP) technology in buffalo. It is known that proteolytic enzymes are involved in different steps of the fertilization process; among these, a critical role may be played by the plasminogen activator-plasmin system. It has been demonstrated that plasmin, the active enzyme of this system, induces acrosome reaction (AR) in bull spermatozoa (Taitzoglou IA et al. 2003 Andrologia 35, 112-116). The aim of this study was to investigate the effect of plasmin on the ability of buffalo sperm to undergo the AR. Frozen- thawed sperm from 4 buffalo bulls were treated by swim-up and incubated with 0.01 mM heparin for 4 h. At 0, 2, and 4 h, aliquots of spermatozoa were exposed for 10 min to 60 μg mL-1 of lysophosphatidylcholine (LPC), as positive control, and to 0.01 μg mL-1 of plasmin. This concentration was chosen after a preliminary dose-response trial. Another sample from each treatment was incubated with IVF medium (negative control). After 10 min, sperm motility was evaluated and sperm were fixed in 37% formaldehyde and stained with trypan blue-Giemsa for subsequent microscopic examination. The total number of sperm counted, over 3 replicates, was 1269 for the negative control, 1293 for LPC, and 1238 for plasmin, equally distributed among incubation times. Differences among groups were analyzed by chi-square test. After swim-up, acrosomal loss was observed only in 4% of the sperm. The addition of 0.01 μg mL-1 of plasmin for 10 min to buffalo spermatozoa at time 0 significantly (P < 0.01) enhanced (23%) AR compared with the control (7.8%), with the same efficiency of LPC (17.1%). After 2 h of incubation with heparin, both plasmin and LPC increased the AR compared to the control (24.4, 20.1, and 14.0%, respectively; P < 0.01). After 4 h, plasmin gave higher percentages of AR (27.2%) compared to both the control (21.0%; P < 0.05) and LPC (19.2%; P < 0.01). Another interesting result is the improved motility recorded with plasmin compared to both the control and LPC groups at 2 h of incubation (90, 75, and 75%, respectively; P < 0.05) and at 4 h of incubation (75, 60, and 60%, respectively; P < 0.05). Finally, no differences in sperm viability were observed between plasmin and the control, whereas a decreased viability was found when LPC was used at 0 h (96.2, 95.0, and 89.0%, respectively, for plasmin, control, and LPC; P < 0.05), at 2 h (85.0, 87.5, and 77.0%, respectively, for plasmin, control, and LPC; P < 0.01), and at 4 h (85.0, 93.3, and 81.1%, respectively, for plasmin, control, and LPC; P < 0.01). In conclusion, we found that addition of plasmin to capacitated sperm increases the percentage of acrosome-reacted spermatozoa and improves motility. Our results suggest that plasmin may play a role in events surrounding fertilization and suggest to evaluate in further studies whether the addition of plasmin during IVF improves the IVEP efficiency in buffalo.


2011 ◽  
Vol 29 (4_suppl) ◽  
pp. 589-589
Author(s):  
S. Yuki ◽  
H. Nakatsumi ◽  
M. Tateyama ◽  
Y. Uehata ◽  
M. Kudo ◽  
...  

589 Background: Indisetron is a 5-HT3 receptor antagonist which shows also 5-HT4 antagonistic activity, that had approved in 2004 by Japan's PMDA. There are no recommendations of prophylactic regimens for preventing nausea and vomiting induced by FOLFOX therapy. To explore the optimal dose period of indisetron tablets during mFOLFOX6, we designed the study to compare the antiemetic efficacy and safety of 3-day regimen of indisetron with a single dose regimen. Methods: Advanced colorectal cancer patients who were treated with mFOLFOX6 (+/- bevacizumab) as first-line chemotherapy were enrolled in this study. They were randomly assigned to Group A (3-day of indisetron) or Group B (a single dose of indisetron). Dexamethazone (8mg) was also administered intravenously in both groups before administering of oxaliplatin. The follow-up period was 5 days from the start of chemotherapy. The primary endpoint was complete protection from vomiting, and secondary endpoints were complete protection from nausea, no use of rescue therapy, and severe adverse events. Results: Of 45 patients enrolled in this trial, 42 (93.3%) were assessable. The proportions of patients with complete protection from vomiting were 85.7% in Group A, and 81.0% in Group B (p=1.000; Fisher's exact test). The proportions of patients with complete protection from nausea were 47.6% in each group (p=1.000; chi-square test). The no rescue therapy rates were 66.7% in Group A, and 57.1% in Group B (p=0.525; chi-square test). No severe adverse events were observed in both groups. Conclusions: We suggested that the efficacy of a single dose of indisetron might be equivalent of 3-day regimen for preventing from nausea and vomiting induced by mFOLFOX6. [Table: see text]


Author(s):  
Obaid Ul Haseeb ◽  
Haris Rashid ◽  
Afrin Ahmed ◽  
Mir Arsalan Ali ◽  
Shakil Alam ◽  
...  

Background: Laparoscopic surgery or "minimally invasive" surgery is a type of specialist surgery. The most crucial, demanding, and risky part of the laparoscopy is the creation of pneumoperitoneum. The two most prevalent methods for creating a pneumoperitoneum are the closed and open approaches. Despite the fact that there is no universal consensus on the best approach to gain access to the peritoneal cavity in order to create a pneumoperitoneum. The aim of present study was to compare the operative time and post-operative outcome associated with closed technique and open classic technique. Methods: This randomized controlled trial was carried out by recruiting patients presented in surgery department of Ziauddin Hospital North campus. The samples were divided into two equal groups A and B. Group A was operated for Laparascopic abdominal surgery by open technique while group B operated through closed technique. Effectiveness of procedures was measured by number of complications occurred during and after surgery. Chi-square test and independent T-test were applied for association. P-value of < 0.05 was considered as significant. Results: Mean age was found to be 45.5±16 years and mean weight was 68±10.5 kilograms. Mean time of operations was 84.5±18.5 minutes. 60 (69.8%) of patients included underwent the laparoscopic cholecystectomy while 15 (17.4%) patients had laparoscopic appendectomy. 13/86: 15.1% of patients had the minor complication including 12/43: 27.9% in group B and 1/43: 2.3% in group A. The comparative analysis between the two groups in terms of effectiveness of either method compared by means of development of the complications was found to be highly significant with p value 0.002. Conclusion: The open approach to laparoscopic entrance has been linked to fewer surgical problems than the closed approach.


2017 ◽  
Vol 24 (11) ◽  
pp. 1749-1754
Author(s):  
Muhammad Abdullah ◽  
Zaheer Ahmed ◽  
Hassan Fareed

The aim on the study was to compare the efficancy of salmeterol and formoterolin persistent asthama. Study Design: Randomized-Controlled-Trial(RCT). Setting: Departmentof Medicine, Allied Hospital, Faisalabad. Period: June 2014 to December 2014. Methodology:Patients of both genders with ages between 18 and 70 years having persistent bronchial asthmawhile Pregnant or lactating mothers, patients with upper or lower respiratory tract infections,acute asthma exacerbations within 4 weeks of first visit, Oral corticosteroids within 4 weeks ordepot steroids within 12 weeks of first visit and Smoking history of more than 10 pack yearswere excluded from study. Patients were randomly divided into two groups (Group A & GroupB) using computer generated random number table. Salmeterol/Fluticasone combination wasgiven to group A with a dose of 50/250μg, 2 actuations with ABEL SPACER DEVICE twice aday for a period of 24 weeks. Formoterol/Budesonide combination was given to group B with adose of 400/6μg with Rotahaler twice a day. Follow up was done by patient’s outdoor visits at6th,12th,18h and 24th week. Results: 180 patients were enrolled in the study. 79 (44%) weremales and 101 (56%) were females. Mean age of study population was 45.25+13.382 years.Patients in Group B experienced lesser number of exacerbations than patients in Group B.Group B showed better response to treatment than Group A using chi square test. (P-Value0.001). Conclusion: It has been concluded that budesonide/Formoterol is more effective incontrolling asthma symptoms than fluticasone/Salmeterol.


2021 ◽  
Vol 71 (Suppl-3) ◽  
pp. S437-39
Author(s):  
Muhammad Sohail Babar Niazi ◽  
Khawar Kamal ◽  
Zaheer Ul Hassan ◽  
Aisha Qaiser ◽  
Shahid Farooq Khattak ◽  
...  

Objective: To carry out the comparison of the effectiveness of Epleys manoeuvre and Vestibular Sedative in the management of benign paroxysmal positional vertigo. Study Design: Quasi experimental study. Place and Duration of Study: Department of ENT, Combined Military Hospital Peshawar, from Jul 2019 to Dec 2020. Methodology: The study was conducted on 60 patients who were diagnosed Benign Paroxysmal Positional Vertigo by Dix Hallpike test. The patients were divided into 2 groups of 30 each. Group A was treated with vestibular sedative and group B was treated with Epleys manoeuvre. The examination of all patients was done on day 3, day 7 and day 14 and the results were documented. Results: The study revealed that the 30 cases which were managed by Epleys manoeuvre, 27 cases were completely symptom free after 14 days. Meanwhile from the 30 patients who were treated by oral vestibular sedation, only 5 patients had total recovery on 14th day. Chi square test was used to compare the results. The results showed that Epleys manoeuvre has greater efficacy in the treatment of Epleys man oeuvre than vestibular sedative. Conclusion: Epleys manoeuvre is more effective for the management of benign paroxysmal positional vertigo than vestibular sedative.


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