Gene Amplification Permits Minimally Invasive Analysis of Fish Mitochondrial DNA

Introduction Mitochondrial DNA (mtDNA) is gaining increasing interest as a marker of cellular damage and could also act as an inflammatory mediator in cardiopulmonary bypass induced postoperative inflammatory response. Although minimally invasive heart valve surgery reportedly reduces inflammation, the mtDNA and cytokine profile in this context remains unclear. Materials and Methods Here, we report a prospective series of 40 elderly patients with aortic stenosis who underwent bioprosthetic aortic valve replacement (AVR) through upper ministernotomy with either a sutureless (n = 20) or a conventional (n = 20) valve. Primary end points included serial plasma levels of mtDNA (T1: at baseline; T2: 4 hours after surgery; and T3: 24s hour after surgery), cytokines (interleukin-6 [IL-6], tumor necrosis factor-α [TNF-α]), and myocardial necrosis biomarkers (MNBs), whereas secondary end points included clinical and echocardiographic data. Results Significant increases in the postoperative plasma levels (T2) of mtDNA, cytokines, and MNBs were observed in all patients. The postoperative plasma levels of mtDNA, TNF-α, and MNBs showed no significant differences between the treatment groups, although there was a trend toward lower levels in the sutureless group. The decreases in aortic cross-clamp and cardiopulmonary bypass times seen in the sutureless group were associated with significant lower postoperative levels (T2 and T3) of IL-6. Conclusion AVR through upper ministernotomy was associated with a significant increase in postoperative plasma levels of mtDNA and cytokines. There was no difference in the mtDNA levels between the sutureless and conventional valve groups, suggesting a similar level of inflammation in both groups. However, the shorter operation time observed in the sutureless valve group was associated with significantly lower postoperative levels of IL-6, indicating potential clinical benefits.

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Multiple populations of deleted mitochondrial DNA detected by a novel gene amplification method

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(89)92362-0 ◽

1989 ◽

Vol 162 (2) ◽

pp. 664-672 ◽

Cited By ~ 48

Author(s):

Wataru Sato ◽

Masashi Tanaka ◽

Kinji Ohno ◽

Tomoko Yamamoto ◽

Goro Takada ◽

...

Keyword(s):

Mitochondrial Dna ◽

Gene Amplification ◽

Multiple Populations ◽

Novel Gene ◽

Amplification Method

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Evaluation of the dark field method for large association of spread DNA

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081814 ◽

1978 ◽

Vol 36 (2) ◽

pp. 190-191

Author(s):

Douglas C. Barker

Keyword(s):

Electron Microscopy ◽

Molecular Weight ◽

Mitochondrial Dna ◽

Extraction Method ◽

Field Method ◽

Dark Field ◽

Kinetoplast Dna ◽

Field Electron ◽

Field Electron Microscopy ◽

Dark Field Electron

A number of satisfactory methods are available for the electron microscopy of nicleic acids. These methods concentrated on fragments of nuclear, viral and mitochondrial DNA less than 50 megadaltons, on denaturation and heteroduplex mapping (Davies et al 1971) or on the interaction between proteins and DNA (Brack and Delain 1975). Less attention has been paid to the experimental criteria necessary for spreading and visualisation by dark field electron microscopy of large intact issociations of DNA. This communication will report on those criteria in relation to the ultrastructure of the (approx. 1 x 10-14g) DNA component of the kinetoplast from Trypanosomes. An extraction method has been developed to eliminate native endonucleases and nuclear contamination and to isolate the kinetoplast DNA (KDNA) as a compact network of high molecular weight. In collaboration with Dr. Ch. Brack (Basel [nstitute of Immunology), we studied the conditions necessary to prepare this KDNA Tor dark field electron microscopy using the microdrop spreading technique.

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Properties of Isolated Mitochondria of Agaricus Bisporus: Their Relationship to Dormancy and the Germination of Spores

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072733 ◽

1973 ◽

Vol 31 ◽

pp. 458-459

Author(s):

K. S. McCarty ◽

R. F. Weave ◽

L. Kemper ◽

F. S. Vogel

Keyword(s):

Mitochondrial Dna ◽

Ethidium Bromide ◽

Microscopic Examination ◽

Agaricus Bisporus ◽

Osmotic Shock ◽

Electron Microscopic Examination ◽

Electron Microscopic ◽

Isolated Mitochondria ◽

Dna Strands ◽

Cytoplasmic Ribosomes

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.

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Genetics of Hearing Loss: Contributions of Mitochondrial DNA

Perspectives on Hearing and Hearing Disorders in Childhood ◽

10.1044/hhdc12.1.2 ◽

2002 ◽

Vol 12 (1) ◽

pp. 2-5

Author(s):

Jill L. Elfenbein

Keyword(s):

Hearing Loss ◽

Mitochondrial Dna

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Considerations in Dysphagia Management Following Esophagectomy

Perspectives of the ASHA Special Interest Groups ◽

10.1044/persp1.sig13.169 ◽

2016 ◽

Vol 1 (13) ◽

pp. 169-176

Author(s):

Lisa M. Evangelista ◽

James L. Coyle

Keyword(s):

Esophageal Cancer ◽

Minimally Invasive ◽

Minimally Invasive Surgical Procedures ◽

Esophageal Resection ◽

Minimally Invasive Surgical ◽

Operative Complications ◽

Swallowing Difficulties ◽

Epidemiological Impact ◽

Invasive Techniques ◽

Management Of Dysphagia

Esophageal cancer is the sixth leading cause of death from cancer worldwide. Esophageal resection is the mainstay treatment for cancers of the esophagus. While curative, surgical resection may result in swallowing difficulties that require intervention from speech-language pathologists (SLPs). Minimally invasive surgical procedures for esophageal resection have aimed to reduce morbidity and mortality associated with more invasive techniques. Both intra-operative and post-operative complications, regardless of the surgical approach, can result in dysphagia. This article will review the epidemiological impact of esophageal cancers, operative complications resulting in dysphagia, and clinical assessment and management of dysphagia pertinent to esophageal resection.

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