102 REPEATED OVARIAN STIMULATIONS BY EXOGENOUS GONADOTROPIN COULD ALTER ATP CONTENT AND MITOCHONDRIAL DISTRIBUTION IN MOUSE OOCYTES

2013 ◽  
Vol 25 (1) ◽  
pp. 198
Author(s):  
Y. Kameyama ◽  
G. Shimoi ◽  
S. Kubo ◽  
R. Hashizume

Ovarian stimulation (OS) by exogenous gonadotropin enhances the availability of mammalian oocytes but compromises their developmental competence (Combelles and Albertini 2003 Biol. Reprod. 68, 812–821). Recently, several studies have reported mitochondrial function-related abnormalities in oocytes after single and repeated OSs. Because mitochondria can directly influence fertilization outcomes (El Shourbagy et al. 2006 Reproduction 131, 233–245), this study aimed to determine the relationship between mitochondria-related parameters and developmental competence on the basis of ATP content, mitochondrial DNA (mtDNA) copy number, mitochondrial distribution, and IVF results for oocytes after repeated OSs. Ovulated oocytes were recovered from ICR female mice treated with 7.5 IU of eCG and 5 IU of hCG at an interval of 48 h in 1 (control) to 3 stimulation cycles, which were performed at intervals of either 5 or 10 d (n = 15–25 in each treatment group). The ATP content in oocytes was determined using a luminometer and commercial kits (BacTiter-Glo; Promega, WI, USA; n = 15–29 in each treatment group). The mtDNA copy number in oocytes was quantified by performing absolute quantification with real-time PCR (n = 4–8 in each treatment group). Mitochondrial distribution in oocytes stained by rhodamine123 was observed under a confocal microscope (n = 12–26 in each treatment group). These analyses were performed only for morphologically normal oocytes. The data were analyzed by one-way ANOVA, followed by Fisher’s least significant difference, or by the chi-square test. Some mice did not ovulate in the third stimulation cycle for both intervals (5-d interval, 32%; 10-d interval, 80%). The mean numbers of ovulated oocytes gradually decreased with progression of the stimulation cycles. The ATP content of the oocytes significantly decreased both in the second and third stimulation cycles, performed with a 5-d interval (control, 1.038 ± 0.117 pmol; second cycle, 0.852 ± 0.189 pmol; third cycle, 0.932 ± 0.272 pmol). The mean mtDNA copy number in oocytes did not change significantly but varied widely in the third stimulation cycle (control, 146 000 ± 21 000; 5-d interval, 135 000 ± 35 000; 10-d interval: 148 000 ± 50 000; mean ± SD). The mitochondrial staining patterns were classified into homogeneous, aggregation, and perinuclear accumulation. The rates of aggregation and perinuclear accumulation increased after repeated stimulation cycles. The blastocyst rates did not significantly differ among the treatment groups after IVF (75–82%). Repeated OSs not only decreased the number of ovulated oocytes but also caused changes related with mitochondrial function, even in the morphologically normal ovulated oocytes. Translocation of active mitochondria, which are associated with energy production, has some functional correlation with successful pre-implantation development (Suzuki et al. 2006 J. Mamm. Ova. Res. 23, 128–134). Changes in mitochondrial distribution might compensate for the negative effect of the decrease in ATP content and low mtDNA copy number after repeated OSs to help reach the blastocyst stage.

Author(s):  
Eva C. Böckmann ◽  
E. S. Debus ◽  
R. T. Grundmann

Abstract Purpose The publication activity of 38 German general/visceral surgery university departments, documented by first or last authorship from staff surgeons (chief and consultants), was evaluated. Methods The observation period extended from 2007 to 2017 and all PubMed-listed publications were considered. Impact factor (IF) was evaluated through the publishing journal’s 5-year IF in 2016, as was the IF for each individual publication. Ranking was expressed in quartiles. Results The staff surgeons of the 38 departments comprised 442 surgeons, of which only 351 (79.4%) were active as first or last authors. Four thousand six hundred and ninety-nine publications published in 702 journals were recorded. The four leading departments in publication number published as much as the last 20 departments (1330 vs. 1336 publications, respectively). The mean of the first (most active) department quartile was 19.6 publications, the second 15.4, the third 11.0, and the last quartile 7.6 per publishing surgeon. The total cumulative impact factor was 14,130. When examining the mean number of publications per publishing surgeons per the 10 year period, the mean of the first quartile was 57.9 cumulative IF, the second 45.0, the third 29.5, and the fourth quartile 17.1. With 352 (7.5%) publications, the most frequently used journal was Chirurg, followed by Langenbeck’s Archives of Surgery with 274 (5.8%) publications. Pancreas-related topics led in terms of publication number and IF generated per individual publication. Conclusion A significant difference in publication performance of individual departments was apparent that cannot be explained by staff number. This indicates that there are as yet unknown factors responsible for minor publication activity in many university departments.


Parasitology ◽  
1941 ◽  
Vol 33 (1) ◽  
pp. 40-46 ◽  
Author(s):  
A. J. Haddow

1. Isolated unmated female body-lice were worn in pillboxes between the skin and the clothes. They were kept constantly on the body but, by a simple device, groups of ten were permitted feeding periods of different length. These groups were fed for 4, 8, 12, 16, 20 and 24 hr. per day respectively. Another group of ten were never allowed to feed after the last moult.2. Some of the figures for egg yield were high. Lice in the 24 hr. group were able to maintain a rate of ten eggs per day for 4−5 days at a time.3. No significant difference in longevity or rate of egg-laying was found to exist between the 12, 16, 20 and 24 hr. groups nor between the 4 and 8 hr. groups but a pronounced and significant difference exists between the 8 and 12 hr. groups. Below 12 hr. there is a sharp fall in longevity and rate of egg production. The unfed group all died, without laying, on the third day.4. The rate of laying as shown by the mode increases progressively with increase in time allowed daily for feeding.5. With regard to the mean eggs per louse the position is less clear. It is felt that the 24 hr. group may differ significantly from the 12, 16 and 20 hr. groups but this is uncertain.


2018 ◽  
Vol 5 (10) ◽  
pp. 2726-2732
Author(s):  
Masoum Khoshfetrat ◽  
Sima Davarpanah ◽  
Aliakbar Keykha

Background: Co-administration of drugs with synergistic effects is considered as one of the methods to increase the effectiveness of intrathecal anesthesia and to reduce the need for injectable analgesics. The purpose of this study was to investigate the efficacy of intrathecal midazolam on enhancing the analgesic effect of fentanyl in patients undergoing lower limb surgery. Materials: The present double-blinded clinical trial was conducted on 90 candidate patients undergoing lower limb orthopedic surgery in 2017 at Khatam-Al-Anbiya Hospital in the city of Zahedan, Iran. To this end, the patients were selected via convenience sampling method and then randomly divided into three groups. Afterwards, 3 cc of 0.5% hyperbaric Marcaine was injected intrathecally in the first group, 3 cc of Marcaine + 25 ug of fentanyl was administered to the second group, and 3 cc of Marcaine + 25 ug of fentanyl + 1 mg of midazolam was administered to the third group (the final volume of all three syringes was brought up to 3.7 cc with normal saline). The duration of sensory nerve block and those of motor block and analgesia, as well as changes in vital signs, were then measured in these groups. Moreover, the data were analyzed using SPSS Statistics (Version 22.0) through Chi-square test and one-way analysis of variance (ANOVA). Results: The mean age and gender distribution of the patients in the three study groups did not differ significantly. The mean duration of sensory nerve block was equal to 98.6+/-11.1 min in the first group, 142.2+/-12.4 min in the second group, and 174.3+/-10.9 min in the third group; all showed a statistically significant difference (P=0.0001, P=0.001, P=0.000).The mean duration of analgesia was also reported to be 204.43+/-0.3 min in the first group, 323.62+/-7.4 min in the second group, and 526.22+/-0.9 min in the third group, indicating a statistically significant difference between the three study groups (P=0.0001, P=0.000, P=000). Conclusion: The results of this study demonstrated that adding midazolam to intrathecal fentanyl could significantly increase the duration of sensory nerve block and that of post-operative analgesia.  


Author(s):  
Sophia Bam ◽  
Erin Buchanan ◽  
Caitlyn Mahony ◽  
Colleen O’Ryan

Autism spectrum disorder (ASD) is a complex disorder that is underpinned by numerous dysregulated biological pathways, including pathways that affect mitochondrial function. Epigenetic mechanisms contribute to this dysregulation and DNA methylation is an important factor in the etiology of ASD. We measured DNA methylation of peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1α), as well as five genes involved in regulating mitochondrial homeostasis to examine mitochondrial dysfunction in an ASD cohort of South African children. Using targeted Next Generation bisulfite sequencing, we found differential methylation (p < 0.05) at six key genes converging on mitochondrial biogenesis, fission and fusion in ASD, namely PGC-1α, STOML2, MFN2, FIS1, OPA1, and GABPA. PGC-1α, the transcriptional regulator of biogenesis, was significantly hypermethylated at eight CpG sites in the gene promoter, one of which contained a putative binding site for CAMP response binding element 1 (CREB1) (p = 1 × 10–6). Mitochondrial DNA (mtDNA) copy number, a marker of mitochondrial function, was elevated (p = 0.002) in ASD compared to controls and correlated significantly with DNA methylation at the PGC-1α promoter and there was a positive correlation between methylation at PGC-1α CpG#1 and mtDNA copy number (Spearman’s r = 0.2, n = 49, p = 0.04) in ASD. Furthermore, DNA methylation at PGC-1α CpG#1 and mtDNA copy number correlated significantly (p < 0.05) with levels of urinary organic acids associated with mitochondrial dysfunction, oxidative stress, and neuroendocrinology. Our data show differential methylation in ASD at six key genes converging on PGC-1α-dependent regulation of mitochondrial biogenesis and function. We demonstrate that methylation at the PGC-1α promoter is associated with elevated mtDNA copy number and metabolomic evidence of mitochondrial dysfunction in ASD. This highlights an unexplored role for DNA methylation in regulating specific pathways involved in mitochondrial biogenesis, fission and fusion contributing to mitochondrial dysfunction in ASD.


2019 ◽  
Vol 31 (3) ◽  
pp. 570 ◽  
Author(s):  
W. Karin Hendriks ◽  
Silvia Colleoni ◽  
Cesare Galli ◽  
Damien B. B. P. Paris ◽  
Ben Colenbrander ◽  
...  

Intracytoplasmic sperm injection is the technique of choice for equine IVF and, in a research setting, 18–36% of injected oocytes develop to blastocysts. However, blastocyst development in clinical programs is lower, presumably due to a combination of variable oocyte quality (e.g. from old mares), suboptimal culture conditions and marginal fertility of some stallions. Furthermore, mitochondrial constitution appears to be critical to developmental competence, and both maternal aging and invitro embryo production (IVEP) negatively affect mitochondrial number and function in murine and bovine embryos. The present study examined the onset of mitochondrial (mt) DNA replication in equine embryos and investigated whether IVEP affects the timing of this important event, or the expression of genes required for mtDNA replication (i.e. mitochondrial transcription factor (TFAM), mtDNA polymerase γ subunit B (mtPOLB) and single-stranded DNA binding protein (SSB)). We also investigated whether developmental arrest was associated with low mtDNA copy number. mtDNA copy number increased (P<0.01) between the early and expanded blastocyst stages both invivo and invitro, whereas the mtDNA:total DNA ratio was higher in invitro-produced embryos (P=0.041). Mitochondrial replication was preceded by an increase in TFAM but, unexpectedly, not mtPOLB or SSB expression. There was no association between embryonic arrest and lower mtDNA copy numbers.


2019 ◽  
Vol 20 (23) ◽  
pp. 6086 ◽  
Author(s):  
Meng Xu ◽  
Qing Ma ◽  
Chunlan Fan ◽  
Xue Chen ◽  
Huiming Zhang ◽  
...  

This study aimed to evaluate whether ginsenosides Rb1 (20-S-protopanaxadiol aglycon) and Rg1 (20-S-protopanaxatriol aglycon) have mitochondrial protective effects against oxygen-glucose deprivation/reoxygenation (OGD/R)-induced injury in primary mouse astrocytes and to explore the mechanisms involved. The OGD/R model was used to mimic the pathological process of cerebral ischemia-reperfusion in vitro. Astrocytes were treated with normal conditions, OGD/R, OGD/R plus Rb1, or OGD/R plus Rg1. Cell viability was measured to evaluate the cytotoxicity of Rb1 and Rg1. Intracellular reactive oxygen species (ROS) and catalase (CAT) were detected to evaluate oxidative stress. The mitochondrial DNA (mtDNA) copy number and mitochondrial membrane potential (MMP) were measured to evaluate mitochondrial function. The activities of the mitochondrial respiratory chain (MRC) complexes I–V and the level of cellular adenosine triphosphate (ATP) were measured to evaluate oxidative phosphorylation (OXPHOS) levels. Cell viability was significantly decreased in the OGD/R group compared to the control group. Rb1 or Rg1 administration significantly increased cell viability. Moreover, OGD/R caused a significant increase in ROS formation and, subsequently, it decreased the activity of CAT and the mtDNA copy number. At the same time, treatment with OGD/R depolarized the MMP in the astrocytes. Rb1 or Rg1 administration reduced ROS production, increased CAT activity, elevated the mtDNA content, and attenuated the MMP depolarization. In addition, Rb1 or Rg1 administration increased the activities of complexes I, II, III, and V and elevated the level of ATP, compared to those in the OGD/R groups. Rb1 and Rg1 have different chemical structures, but exert similar protective effects against astrocyte damage induced by OGD/R. The mechanism may be related to improved efficiency of mitochondrial oxidative phosphorylation and the reduction in ROS production in cultured astrocytes.


2019 ◽  
Vol 22 (1) ◽  
pp. 139-151 ◽  
Author(s):  
Han Shen ◽  
Man Yu ◽  
Maria Tsoli ◽  
Cecilia Chang ◽  
Swapna Joshi ◽  
...  

Abstract Background Despite increased understanding of the genetic events underlying pediatric high-grade gliomas (pHGGs), therapeutic progress is static, with poor understanding of nongenomic drivers. We therefore investigated the role of alterations in mitochondrial function and developed an effective combination therapy against pHGGs. Methods Mitochondrial DNA (mtDNA) copy number was measured in a cohort of 60 pHGGs. The implication of mtDNA alteration in pHGG tumorigenesis was studied and followed by an efficacy investigation using patient-derived cultures and orthotopic xenografts. Results Average mtDNA content was significantly lower in tumors versus normal brains. Decreasing mtDNA copy number in normal human astrocytes led to a markedly increased tumorigenicity in vivo. Depletion of mtDNA in pHGG cells promoted cell migration and invasion and therapeutic resistance. Shifting glucose metabolism from glycolysis to mitochondrial oxidation with the adenosine monophosphate–activated protein kinase activator AICAR (5-aminoimidazole-4-carboxamide ribonucleotide) or the pyruvate dehydrogenase kinase inhibitor dichloroacetate (DCA) significantly inhibited pHGG viability. Using DCA to shift glucose metabolism to mitochondrial oxidation and then metformin to simultaneously target mitochondrial function disrupted energy homeostasis of tumor cells, increasing DNA damage and apoptosis. The triple combination with radiation therapy, DCA and metformin led to a more potent therapeutic effect in vitro and in vivo. Conclusions Our results suggest metabolic alterations as an onco-requisite factor of pHGG tumorigenesis. Targeting reduced mtDNA quantity represents a promising therapeutic strategy for pHGG.


2019 ◽  
Vol 56 (2) ◽  
pp. 141-145 ◽  
Author(s):  
Maryam GERAVAND ◽  
Parviz FALLAH ◽  
Mojtaba Hedayat YAGHOOBI ◽  
Fatemeh SOLEIMANIFAR ◽  
Malihe FARID ◽  
...  

ABSTRACT BACKGROUND: Colorectal cancer is one of the most commonly diagnosed cancers around the world. One of the factors involved in the development of colorectal cancer is the changes in the normal flora of the intestine. OBJECTIVE: In this study, the mean copy number of Enterococcus faecalis in people with polyps and people with colorectal cancer has been evaluated in comparison with healthy controls. METHODS: In this study, 25 patients with colorectal cancer and 28 patients with intestinal polyps were selected and stool specimens were taken. In addition, 24 healthy individuals were selected as control group. Extraction of bacterial DNA from the stool sample were performed. The molecular methods of PCR for confirmation of standard strain and absolute Real Time PCR (qRT-PCR) method were used to evaluate the number of Enterococcus faecalis in the studied groups. RESULTS: The results of this study indicate that the mean copy number of Enterococcus faecalis in patients with colorectal cancer was 11.2x109 per gram of stool, and in patients with polyps was 9.4x108 per gram of stool. In healthy people, this number was 9x108 per gram of stool. There was a significant difference between the implicit copy numbers in the three groups. (P<0.05). CONCLUSION: Enterococcus faecalis in faecal flora of people with colorectal cancer was significantly higher than those with polyps and healthy people. This could potentially signify the ability of this bacterium to induce colorectal cancer. More studies are needed to prove this theory.


2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Jolanta Opiela ◽  
Joanna Romanek ◽  
Daniel Lipiński ◽  
Zdzisław Smorąg

The objective of the present study was to evaluate the effect of hyaluronan (HA) during IVM on meiotic maturation, embryonic development, and the quality of oocytes, granulosa cells (GC), and obtained blastocysts. COCs were maturedin vitroin control medium and medium with additional 0.035% or 0.07% of exogenous HA. The meiotic maturity did not differ between the analysed groups. The best rate and the highest quality of obtained blastocysts were observed when 0.07% HA was used. A highly significant difference (P<0.001) was noted in the mean number of apoptotic nuclei per blastocyst and in the DCI between the 0.07% HA and the control blastocysts (P<0.01). Our results suggest that addition of 0.035% HA and 0.07% HA to oocyte maturation media does not affect oocyte nuclear maturation and DNA fragmentation. However, the addition of 0.07% HA during IVM decreases the level of blastocysts DNA fragmentation. Finally, our results suggest that it may be risky to increase the HA concentration during IVM above 0.07% as we found significantly higherBaxmRNA expression levels in GC cultured with 0.07% HA. The final concentration of HA being supplemented to oocyte maturation media is critical for the success of the IVP procedure.


2008 ◽  
Vol 78 (2) ◽  
pp. 324-331 ◽  
Author(s):  
Katri Keski-Nisula ◽  
Leo Keski-Nisula ◽  
Hannu Salo ◽  
Kati Voipio ◽  
Juha Varrela

Abstract Objective: To evaluate skeletal and dentoalveolar changes induced by the eruption guidance appliance in the early mixed dentition. Materials and Methods: Pre- and posttreatment cephalometric radiographs of 115 consecutively treated children, 62 boys and 53 girls, were compared with those obtained from a control group of 104 children, 52 boys and 52 girls. Pretreatment radiographs were taken at the deciduous-mixed dentition interphase (T1) and after full eruption of all permanent incisors and first molars (T2). The mean age of the children in both groups was 5.1 years at T1 and 8.4 years at T2. Results: A significant difference between the groups at T2 was found in the mandibular length, midfacial length, and maxillomandibular differential. The increase in mandibular length was 11.1 mm in the treatment group and 7.2 mm in the control group. No differences were found in measurements of maxillary position or size. There was a significant shift toward a Class I relationship in the treatment group. Labial tipping and linear protrusion of the mandibular incisors was evident in the treatment group at T2. There was no effect on the inclination or position of the maxillary incisors. Conclusions: Occlusal correction was achieved mainly through changes in the dentoalveolar region of the mandible. In addition, the appliance enhanced condylar growth resulting in a clinically significant increase in mandibular length. No effect was observed on maxillary position, maxillary size, inclination or protrusion of the maxillary incisors, or facial height.


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