302 VIABILITY OF BOVINE SPERMATOZOA INCUBATED WITH FOREIGN DNA

2006 ◽  
Vol 18 (2) ◽  
pp. 258
Author(s):  
W. B. Feitosa ◽  
M. P. Milazzoto ◽  
M. Rovegno ◽  
L. F. Martins ◽  
J. A. Visintin ◽  
...  
Keyword(s):  
Incubation Period ◽  
Molecular Probes ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Sperm Viability ◽  
Exogenous Dna ◽  
Significant Difference ◽  
Class 1 ◽  
Bovine Spermatozoa ◽  
Over Time

Several studies have been performed over the years to promote the understanding and improvement of the sperm-mediated gene transfer technique. However, little is known about the effect of exogenous DNA in the sperm cells. The aim of this study was to evaluate the effect of the incubation period and exogenous DNA addition on mitochondrial activity and acrosomal status of bovine spermatozoa. Frozen-thawed semen was separated by Percoll gradient (45/90%) at 600g for 30 min, and the pellet was resuspended and washed by centrifugation in sperm-TALP at 200g for 5 min. The spermatozoa were resuspended in fertilization medium (without heparin) at a concentration of 5 × 106 spermatozoa/mL and incubated at 39°C, 5% CO2 in air with 500 ng pEYFP-Nuc/mL (Clontech, Mountain View, CA, USA) (DNA group) for 1 and 2 h or without DNA (control group) for 0, 1 and 2 h. JC-1 (Molecular Probes; Invitrogen Brasil, Ltd., Sao Paulo, Brazil) was used to determine mitochondrial potential and fluorsecein isothiogyanate (FITC-PSA; Sigma-Aldrich, Brazil) was used to assess acrosomal status. Two microliters of JC-1 (76.5 μM in DMSO) and 50 μL of FITC-PSA (100 μg/mL in DPBS) were added to 150 μL of IVF medium + 5 × 106 spermatozoa/mL; the mixture was incubated at 25°C for 10 min. DNA incorporation was evaluated by p-EYFP-Nuc PCR amplification. All treatments were repeated ten times and data were analyzed by ANOVA and Tukey test (P < 0.05). The results are described in Table 1. The sperm were classified as: Class 1 (intact acrosome and high mitochondria potential), Class 2 (intact acrosome and low mitochondria potential), Class 3 (reacted acrosome and high mitochondria potential) and Class 4 (reacted acrosome and low mitochondria potential). In both groups, the Class 1 sperm decreased over time, whereas an increase in number was verified for Class 2 sperm. There was no significant difference in numbers among the incubation periods in both groups for Class 3 sperm. However, there was an increase in Class 4 sperm number over time, indicating that the main effect of the incubation period was the loss of mitochondria potential. There was no effect of the exogenous DNA addition on sperm viability in relation to the control group, indicating that the exogenous DNA had no effect on mitochondrial activity and acrosomal status of bovine semen. Table 1. Effect of incubation period and exogenous DNA addition on sperm viability This work was supported by FAPESP 03/10234–7; 03/07456–8.

scholarly journals Effect of Tramadol on Sperm Profile of Male Albino Rats

2019 ◽  
pp. 1-6
Author(s):  
I. S. Esua ◽  
U. U. Uno ◽  
U. B. Ekaluo
Keyword(s):  
Sperm Motility ◽  
Sperm Count ◽  
Sperm Head ◽  
Male Rats ◽  
Control Group ◽  
Albino Rats ◽  
Dependent Manner ◽  
Sperm Viability ◽  
Albino Rat ◽  
Significant Difference

Background and Aim: Tramadol is a potent analgesic effective in the treatment of mild to severe pains. However, the use of the drug can pose a threat to other organs and systems. Therefore, this study evaluated the effect of graded doses of tramadol on sperm profile of male albino rats. Materials and Methods: Eighteen male rats were divided into three groups (A, B and C) using completely randomized design (CRD) with six rats in each group. Rats in group A served as the control group and were given just food and water while groups B and C were given tramadol at 50 and 100 mg/kg body weight (BW) respectively, daily for the period of 65 days. The treatment was administered via oral gavage and at the end of the treatments, the rats were sacrificed. Immediately after sacrifice, a puncture was made in the epididymis with a sterile pin and examined for semen pH. The epididymes were processed for epididymal sperm motility, viability, count and sperm head abnormality. Results: There was no significant difference in the weight of testes and semen pH. Sperm viability, sperm motility, sperm count and weight of epididymes significantly reduced (p<0.05) in tramadol treated animals when compared with the control. Results also indicated statistically significant (p<0.05) increase in sperm head abnormalities in rats treated with tramadol when compared with the control. Conclusion: The results obtained from this study reveal that tramadol has negative effects on weight of epididymes, sperm count, sperm viability, sperm motility and sperm head abnormalities in male albino rat as mammalian models in a dose dependent manner.

113 Effects of Sleep on Intrusive Symptoms and Emotion Reactivity in a Laboratory-Based Film Analogue Study

SLEEP ◽  
2021 ◽  
Vol 44 (Supplement_2) ◽  
pp. A46-A46
Author(s):  
Anna Marie Nguyen ◽  
Rebecca Campbell ◽  
Abigail Vance ◽  
Ellen Leen-Feldner
Keyword(s):  
Sleep Deprivation ◽  
Control Group ◽  
Significant Interaction Effect ◽  
Emotion Reactivity ◽  
Negative Valence ◽  
Significant Difference ◽  
Regulation Strategies ◽  
Acute Sleep Deprivation ◽  
The Impact ◽  
Over Time

Abstract Introduction Recent literature highlights the need to focus on the impact of intrusive symptoms as a possible risk factor for the development and maintenance of PTSD. Cognitive and sleep models also contribute to the further understanding of intrusive symptoms. Further emotion work emphasizes that disgust is an emotion closely associated with the emergence of posttraumatic stress symptomology following traumatic events. Methods This study utilized a film eliciting disgust to examine the effects of acute sleep deprivation on the intensity of intrusive symptoms and emotion reactivity. Forty-nine college students were randomly assigned to sleep as usual or an acute sleep deprivation after watching a disturbing film. It was hypothesized that, relative to the control group, participants who were acutely sleep deprived would report higher frequency of intrusive symptoms and higher negative valence. Results Findings were partially consistent with hypotheses. There were no group or interaction effects on intrusive symptoms, although participants across both groups reported significant decreases in negative valence and intrusive symptoms across the study (F(1, 47) = 10.30, p &lt; 0.01). There was a significant interaction effect between sleep group and self-reported negative valence, where individuals in the sleep deprived group reported significantly higher valence than individuals in the control group, despite significant decreases in negative valence over time (F(1, 48) = 7.869, p &lt; 0.01). Conclusion Possible mechanisms that may contribute to the significant difference in valence may be due to higher order emotion regulation strategies that are compromised due to sleep loss. However, the significant decreases in negative valence and intrusive symptoms over time may be due to methodological factors or the type of sleep manipulation. Further work can address these challenges by using a larger sample size or examining the effects of chronic, partial sleep deprivation. Support (if any):

398 EFFECT OF INCUBATION PERIOD ON TRANSFECTION RATES IN BOVINE SPERMATOZOA

2007 ◽  
Vol 19 (1) ◽  
pp. 314
Author(s):  
M. E. O. A. Assumpção ◽  
W. B. Feitosa ◽  
M. Rovegno ◽  
M. G. Marques ◽  
A. C. Nicacio ◽  
...  
Keyword(s):  
Real Time ◽  
Incubation Period ◽  
Real Time Pcr ◽  
Dna Interaction ◽  
Current Data ◽  
Sperm Cells ◽  
Exogenous Dna ◽  
State College ◽  
Bovine Spermatozoa ◽  
Late Apoptosis

Current data about the kinetics of exogenous DNA interaction with spermatozoa are controversial. The amount of DNA associated with the spermatozoa is proportional to the incubation period. However, a great amount of exogenous DNA, associated or internalized, can activate endonucleasis, which cleaves the exogenous and/or endogenous DNA similarly to apoptosis. The aim of this work was to evaluate the effect of the incubation period of bovine spermatozoa with exogenous DNA on transfection rate and induction of apoptosis, oncosis, or initial oncosis and/or late apoptosis of sperm cells. For that, 5 × 106 spermatozoa/mL were incubated with 500 ng of plasmid pEYFP-Nuc (Clontech, Mountain View, CA, USA) for 60, 90, and 120 min. Sperm cells were then subjected to electrophoresis to remove non-associated exogenous DNA. DNA (endogenous and exogenous) was extracted and real-time PCR was performed to quantify exogenous DNA/spermatozoa association. Each reaction was performed with 12.5 µL of Platinum SYBR Green; 0.5 µL of ROX; 5.6 µL of H2O; 5 µL of sample (1 µg); 0.7 µL of primer sense (5′-ATGGCCGACAAGCAGAAGAAC-3′) and 0.7 µL of primer anti-sense (5′-TGCCGTCCTCGATGTTGTG-3′); 500 nM each. Real-time PCR was conducted for 40 cycles of 95°C for 15 s and 64°C for 1 min. Apoptosis and oncosis analyses were evaluated by flow cytometry with 106 sperm/mL stained with 2 µL of Yo-pro (100 µM) for 20 min and with 10 µL of propidium iodide (6 µM) for 10 min at room temperature. Data were analyzed by MINITAB Realease 14 Statistical Software (Minitab, Inc., State College, PA, USA), subjected to ANOVA, and compared by Fisher's or Tukey's test (P ≤ 0.05) for transfection rate and apoptosis/oncosis, respectively. Linear regression obtained by the amplification of pEYFP-Nuc at different concentrations showed efficiency of R2 = 0.9987. The results summarized in Table 1 show that increase in length of the incubation period increases the amount of exogenous DNA associated with spermatozoa and does not affect the average of live spermatozoa in apoptosis, oncosis, and initial oncosis and/or late apoptosis groups. Table 1.Quantification of exogenous DNA associated with bovine sperm cells and percentages of live, apoptotic, and oncotic spermatozoa This work was supported financially by FAPESP 03/07456-8 and 03/10234-7.

scholarly journals Penerapan Metode Debat dan Model Artikulasi untuk melihat perbandingan kemampuan Berbicara Siswa kelas XI SMK Negeri Palopo

2020 ◽  
Vol 5 (2) ◽  
pp. 38-44
Author(s):  
Besse Herdiana
Keyword(s):  
Experimental Research ◽  
Statistical Techniques ◽  
Control Group ◽  
Product Service ◽  
Significant Difference ◽  
Class 1 ◽  
Post Test ◽  
Sample Technique ◽  
Method Class ◽  
Purposive Sample

This research is an experimental research that aims to compare differences in students' speaking abilities using the debate method and articulation models. This study was designed with a post-test only control group desaign. This research was conducted at SMK Negeri 1 Palopo, with a population of 143 students consisting of four classes. Sampling using purposive sample technique so that the chosen ones are accounting class 1 and accounting class 2. The data obtained is data that explains the comparison of speaking skills between those who use the debate method and those who use the articulation model of students of SMK Negeri 1 Palopo, in this study, the data collected will be analyzed by using descriptive statistical techniques and parametric infrential statistics of the type of independent sample T test that is processed with a computer program Statistics Product Service Solution (SPSS). The results showed that the results of the debating method class completeness category contained 29 students who scored> 75 with a percentage of completeness (94%), and only 2 students who scored ≤ 75 with a percentage of completeness (6%). Likewise with the class using the articulation model there were 29 students who scored> 75 with a percentage of completeness (94%), and only 2 students who scored ≤ 75 with a percentage of completeness (6%). This proves that there is no significant difference between the speaking ability of class XI students of SMK Negeri 1 Palopo using the debate method and the articulation model

scholarly journals Antioxidant Effects of Marigold (Calendula officinalis) Flower Extract on the Oxidative Balance of Bovine Spermatozoa

2019 ◽  
Vol 68 (3-4) ◽  
pp. 92-102
Author(s):  
Filip Benko ◽  
Valentína Palkovičová ◽  
Michal Ďuračka ◽  
Július Árvay ◽  
Norbert Lukáč ◽  
...  
Keyword(s):  
Protein Oxidation ◽  
Quality Parameters ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Calendula Officinalis ◽  
Positive Effects ◽  
Flower Extract ◽  
Bovine Spermatozoa ◽  
Antioxidant Effects

Summary The aim of our study was evaluation of potential antioxidant effects of marigold (Calendula officinalis) extract and assessment of its in vitro impact on the selected quality parameters of bovine spermatozoa. Marigold is medicinal herb from the family Asteraceae native to southern Europe, and it commonly used in pharmacology and medicine. Its well-known positive properties include antioxidant, antibacterial, antiflammatory, antiviral, antifungal, antihelmintic and wound-healing activities. In this study, the flower extracts were subjected to high performance liquid chromatography (HPLC), which identified especially phenolic acids (rosmarinic and chlorogenic acids) and polyphenols (rutin, kaempferol, resveratrol, quercetin and apigenin). These substances are known for their antioxidant activity and protective effects against oxidative stress. For our experiments, 10 samples of semen from sexually mature Holstein bulls were collected on a single day by using an artificial vagina, diluted in physiological saline solution and exposed to solutions with different concentration of marigold flower extract (75, 150 and 300 µg/mL). Selected quality parameters (motility, mitochondrial activity, production of reactive oxygen species – ROS, protein oxidation and lipid peroxidation – LPO) were analyzed after 0, 2 and 24 hours of in vitro culture. The motility evaluation was performed by using the computer-assisted sperm analysis (CASA) method. This method revealed that 75 and 150 µg/mL extract had positive effects and increased the motility (P<0.01) and mitochondrial activity (P<0.0001) of bovine spermatozoa compared to the control group following 2 and 24 hours. This phenomenon was observed also in case of ROS production, protein oxidation and LPO. Marigold extract concentrations of 75 and 150 µg/mL decreased the levels of ROS, protein oxidation and damage to the membranes caused by LPO compared to the control group (P<0.05; P<0.01) at time 2 and 24 hours. At the same time, 300 µg/mL extract exhibited positive, although less significant, effects compared to 75 and 150 µg/mL extracts. The data acquired from our study confirm that 75 and 150 µg/mL of marigold flower extract have positive effects on the motility and mitochondrial activity of bovine spermatozoa, and decrease ROS generation, LPO and protein oxidation in spermatozoa. Based on our results, the flower extract from marigold could be used for protection against oxidative stress in in vitro cultures of male gametes.

scholarly journals Positive effects of trehalose and cysteine on ram sperm parameters

2017 ◽  
Vol 62 (No. 5) ◽  
pp. 245-252 ◽  
Author(s):  
S. Gungor ◽  
C. Ozturk ◽  
AD Omur
Keyword(s):  
Sperm Motility ◽  
Sperm Parameters ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Sperm Viability ◽  
Significant Protection ◽  
Positive Effects ◽  
Liquid Storage ◽  
Acrosome Integrity ◽  
Ram Sperm

The aim of this study was to determine the effects of trehalose and cysteine on sperm motility, viability, mitochondrial activity and acrosome integrity during liquid storage of Merino ram semen. Ejaculates were collected using artificial vaginas from five Merino rams, microscopically evaluated and pooled at 37 °C. The pooled semen samples were diluted in a Tris-based extender, including cysteine (2 mM and 4 mM), trehalose (10 mM and 25 mM) and no antioxidant (control). Diluted semen samples were kept in tubes and cooled from 37 to 5 °C in a cold cabinet, and maintained at 5 °C. Cooled samples were evaluated for sperm motility, viability, mitochondrial activity and acrosome integrity at 0, 24, 48, 72 and 96 h. Extender supplemented with trehalose (10 and 25 mM) and cysteine (2 and 4 mM) led to higher motility in comparison to the control at 24, 48, 72 and 96 h of liquid storage (P &lt; 0.05). Trehalose at the doses of 10 mM, 25 mM and 2 mM cysteine led to higher viability between 24–48–72 h and at 96 h of liquid storage (P &lt; 0.05). Further, 4 mM of cysteine improved sperm viability rates at 24 and 48 h of storage compared to the control group (P &lt; 0.05), and resulted in improved acrosome integrity rates compared to the control group at 72 and 96 h of storage (P &lt; 0.05). Extender supplemented with 10 and 25 mM trehalose at 24 and 72 h and 4 mM cysteine at 24 and 96 h of storage led to higher sperm mitochondrial activity rates when compared to the control group (P &lt; 0.05). In conclusion, the findings of this study show that trehalose and cysteine provided significant protection to ram sperm parameters during liquid storage.

scholarly journals Etlik Piliçlerde Embriyonun Erken ve Geç Gelişim Dönemlerinde Yapılan Yüksek Isıl Uygulamaların Asimetri Üzerine Etkileri

2015 ◽  
Vol 3 (11) ◽  
Author(s):  
Sezai Alkan ◽  
Özgür Barış Birgül
Keyword(s):  
Relative Humidity ◽  
Incubation Period ◽  
Incubation Temperature ◽  
Early Embryogenesis ◽  
Control Group ◽  
Late Embryogenesis ◽  
Significant Difference ◽  
Face Length ◽  
Left And Right ◽  
The Asymmetry

The aim of this study was to determine the effect of thermal manipulations during early and late embryogenesis on asymmetry in terms of sides of shank length, shank width and face length of broilers. Incubation conditions were 37.5°C and 55% relative humidity for control group throughout the incubation period until the 19th days. In the thermally treated eggs during early embryogenesis (8-10 days), incubation temperature was increased to 41°C and relative humidity to 65% for 3 hours (12.00-15.00) on the 8th-10th days of incubation. Also, in the late embryogenesis stage (16-18 days) incubation temperature was increased to 41°C and relative humidity to 65 % for 3 hours (12.00-15.00) on the 16th-18th days of incubation. Total 16 chickens were selected at randomly from all experimental groups to determine the asymmetry. The weekly left and right sides of shank length, shank width and face length of chickens were measured from 7 days of age to 35 days of age, and relative asymmetry values were calculated. There was no significant difference among the groups in point of relative asymmetry. Asymmetry values were reduced due to aging.

scholarly journals Etlik Piliçlerde Embriyonun Erken ve Geç Gelişim Dönemlerinde Yapılan Yüksek Isıl Uygulamaların Asimetri Üzerine Etkileri

2015 ◽  
Vol 3 (11) ◽  
pp. 861
Author(s):  
Sezai Alkan ◽  
Özgür Barış Birgül
Keyword(s):  
Relative Humidity ◽  
Incubation Period ◽  
Incubation Temperature ◽  
Early Embryogenesis ◽  
Control Group ◽  
Late Embryogenesis ◽  
Significant Difference ◽  
Face Length ◽  
Left And Right ◽  
The Asymmetry

The aim of this study was to determine the effect of thermal manipulations during early and late embryogenesis on asymmetry in terms of sides of shank length, shank width and face length of broilers. Incubation conditions were 37.5°C and 55% relative humidity for control group throughout the incubation period until the 19th days. In the thermally treated eggs during early embryogenesis (8-10 days), incubation temperature was increased to 41°C and relative humidity to 65% for 3 hours (12.00-15.00) on the 8th-10th days of incubation. Also, in the late embryogenesis stage (16-18 days) incubation temperature was increased to 41°C and relative humidity to 65 % for 3 hours (12.00-15.00) on the 16th-18th days of incubation. Total 16 chickens were selected at randomly from all experimental groups to determine the asymmetry. The weekly left and right sides of shank length, shank width and face length of chickens were measured from 7 days of age to 35 days of age, and relative asymmetry values were calculated. There was no significant difference among the groups in point of relative asymmetry. Asymmetry values were reduced due to aging.

The effects of magnetic separation on cryopreserved bovine spermatozoa motility, viability and cryo-capacitation status

Zygote ◽  
2012 ◽  
Vol 22 (3) ◽  
pp. 378-386 ◽  
Author(s):  
S.S.M. Faezah ◽  
F.M.Y Zuraina ◽  
J.H.F. Farah ◽  
O. Khairul ◽  
N.I. Hilwani ◽  
...  
Keyword(s):  
Annexin V ◽  
Control Group ◽  
Preparation Technique ◽  
Sperm Preparation ◽  
Spermatozoa Motility ◽  
Male Gametes ◽  
Significant Difference ◽  
Long Time ◽  
Bovine Spermatozoa ◽  
Magnetic Activated Cell Sorting

SummaryCryopreservation is a technique used to preserve cells for long-time storage. It is widely used in agriculture to store male gametes in liquid nitrogen. The aim of this study was to determine the optimum thawing temperature and time for samples subjected to annexin V magnetic-activated cell sorting (AnMACS) as the sperm preparation technique. Pooled semen samples from three ejaculates were divided into two groups. The treatment group was subjected both to AnMACS and to being cryopreserved, whilst the control group was cryopreserved directly without MACS. Post-thaw analysis was carried out for samples thawed at either 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s or 80°C for 5 s. Sperm kinematics, viability and capacitation status were determined for samples subjected to all thawing temperatures described. Results showed that thawing at 37°C for 13 s for MACS-processed samples was a superior option compared with other thawing procedures; there was a significant difference in P < 0.05 values for curvilinear velocity (VCL μm/s) and sperm straightness (STR %) when samples were thawed at 40°C for 7 s, with fewer capacitated spermatozoa (P < 0.05) when samples were thawed at 37°C for 30 s, 40°C for 7 s or 60°C for 6 s. Hence, we can speculate that the use of AnMACS as the sperm preparation technique can somehow enhance sperm cryosurvival rate after cryopreservation, however the fertilization potential of these cells has yet to be determined.

scholarly journals Progressive contralateral hippocampal atrophy following Temporal Lobe Epilepsy Surgery (TLS)

2015 ◽  
Vol 42 (S1) ◽  
pp. S26-S27
Author(s):  
CA Elliott ◽  
C Yasuda ◽  
L Concha ◽  
M Liu ◽  
M Wheatley ◽  
...  
Keyword(s):  
Temporal Lobe Epilepsy ◽  
Temporal Lobe ◽  
Time Course ◽  
Hippocampal Volume ◽  
Control Group ◽  
Hippocampal Atrophy ◽  
Subject Design ◽  
Significant Difference ◽  
Delayed Scan ◽  
Over Time

Background: Temporal Lobe Epilepsy is associated with bilateral gray (GM) and white matter (WM) loss. After surgical treatment progressive bilateral temporal and extra-temporal WM change occur, however, less is known regarding post-operative GM change. We set out to measure contralateral hippocampal volume (CHV) following TLS. Methods: 1.5T-3D-1mm-isotropic-MPRAGE scans in 26 TLE patients and 3 controls in two groups: longitudinal (n=10)(imaged POD1,2,3,6,60,120 and >360d) and single post-operative scan (n=16). Manual volumetry protocols. Results: We find significant CHV atrophy at delayed scan relative to baseline (mean atrophy 26.8%). In the longitudinal group there is significant and progressive atrophy from baseline to POD4-8 (72.6+/-6.5%), POD60-360 (69.7+/-12.3%) and >360 (58.5+/-10.6%). No significant atrophy in either the control group HV or contralateral CV over time. No significant difference in mean HV at the most delayed exam for surgery type (p=0.13) or side (p=0.24). Conclusions: We find a statistically significant CHV atrophy following surgery which is progressive over time. Our longitudinal within-subject design describes the time course and extent more fully than previous work. Caudate analysis indicates that early CHV atrophy is not due to global atrophy following brain surgery but rather may be due to deafferentation and deefferentation. Finally, we find no significant difference in atrophy when analyzed by surgical approach or surgical side.

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