The effect of cryopreservation on mitochondrial function in freshwater mussel tissue samples (Bivalvia: Unionida)

Rational: Heart disease is the leading cause of death worldwide and abnormalities in mitochondrial function are increasingly recognized in association with cardiomyopathy, heart failure, endothelial dysfunction and coronary artery disease (CAD). However the direct contribution and mechanism of the mitochondrial dysfunction on the development of CAD is not fully determined. We have recently shown a critical role of TERT, the catalytic subunit of telomerase, as a regulator of mitochondrial integrity in the microcirculation. We observed increased expression of the dominant negative splice variant of TERT ( β -del) in the Left Ventricle from subjects with CAD. Therefore, we hypothesize that TERT ( β -del) decreases mitochondrial telomerase activity in the human heart resulting in mitochondrial damage that contributes to an environment that promotes the development of CAD . Methods: Fresh and frozen tissue samples of discarded heart tissue from subjects with and without CAD were used. Protein, cell lysate or mitochondria were isolated using standard techniques. Mitochondrial DNA, levels of NAD+ and ATP as well as mitochondrial oxidative phosphorylation were evaluated. Results: PCR analysis revealed an increased frequency of mitochondrial common deletion, an established marker for mitochondrial abnormalities (0.9±0.2 in CAD; vs 1.5±0.2 in non-CAD; N=8; P<0.05). NAD+ and ATP levels were significantly decreased in CAD subjects compared to non-CAD (291±62 and 0.5±0.1 RLU/mg protein in CAD vs. 4203±336 and 84.1±24.8 pmol/mg protein in non-CAD respectively; N=15; P<0.005). Decrease respiration control index (RCI) in the presence of either complex I substrate K (+)-pyruvate/malate (PM) or complex II substrate K (+)-succinate (SUC) was observed in tissue form subjects with CAD (KPM-RCI: 2.9±1.3; SUC-RCI: 7.6± 1.9 in CAD vs KPM-RCI: 8.5±1.9; SUC-RCI: 19.1± 8.3 in non-CAD; N=3; P<0.05) Conclusions: Together these results point to significantly impaired mitochondrial function in subjects with CAD that are associated with decreased in mitochondrial telomerase activity.

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Use of the Zebra Mussel (Dreissena polymorpha) as a Bioindicator for Aromatic Hydrocarbons in Hamilton Harbour

Water Quality Research Journal ◽

10.2166/wqrj.2000.004 ◽

2000 ◽

Vol 35 (1) ◽

pp. 59-72 ◽

Cited By ~ 3

Author(s):

Chris Marvin ◽

Laurie Allan ◽

Douglas Bryant ◽

Brian McCarry

Keyword(s):

Aromatic Hydrocarbons ◽

Dreissena Polymorpha ◽

Metabolic Activation ◽

Gas Chromatography Mass Spectrometry ◽

Gel Chromatography ◽

Mussel Tissue ◽

Tissue Samples ◽

Hamilton Harbour ◽

Freeze Dried ◽

Polycyclic Aromatic

Abstract Three methods for the extraction of polycyclic aromatic hydrocarbons (PAHs) from zebra mussels (Dreissena polymorpha) sampled from Hamilton Harbour were compared. Replicate freeze-dried mussel tissue samples were extracted using acid digestion, tissue homogenization (mechanical extraction) and ultra-sonication. Each extract was submitted to a cleanup procedure (alumina chro-matography and Sephadex LH20 gel chromatography), followed by analysis using gas chromatography-mass spectrometry (GC-MS). The three extraction methods were equally efficient, based on a statistical comparison of mean concentrations of individual PAHs. Mussel extracts, when subjected to bioassays with Salmonella typhimurium strain YG1029 (TA100-like) in the presence of an exogenous metabolic activation system (S9), exhibited significant mutagenic responses; these responses varied with the PAH content of the mussel extracts. Sources of PAHs in mussel extracts were determined by examining the profiles of sulfur-containing polycyclic aromatic compounds (thia-arenes). Comparison of the ratios of certain thia-arenes with ratios in source samples enabled identification of vehicular emissions and coal tar-contaminated sediment as two sources of PAH contamination in Hamilton Harbour.

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HYDROCARBONS DETECTION IN A COASTAL LAGOON IMPACTED BY AN OIL SPILL IN SÃO SEBASTIÃO CHANNEL, SÃO PAULO STATE, BRAZIL1

International Oil Spill Conference Proceedings ◽

10.7901/2169-3358-2005-1-777 ◽

2005 ◽

Vol 2005 (1) ◽

pp. 777-785

Author(s):

Maria Eulália Rocha Carneiro ◽

Rogério Migueis Picado ◽

Cinthia Masumoto ◽

Leonardo Mitidiero Mansor ◽

Märcia Caruso Bícego ◽

...

Keyword(s):

Coastal Lagoon ◽

Petroleum Hydrocarbons ◽

Monitoring Program ◽

Mussel Tissue ◽

Tissue Samples ◽

Total N ◽

Bulk Oil ◽

Lagoon Water ◽

The North ◽

Polycyclic Aromatic

ABSTRACT An operational problem involving the loading arms occurred at São Sebastião Oil Terminal of PETROBRAS, located in the Brazilian Southeastern coast, during the oil transference from the tanker to the Terminal, resulting in the spill of approximately 26.0 m3 of crude Marlin-33 oil on 3 June 2003. The spilled bulk oil was transported by wind-driven currents to the north of São Sebastião Channel where skimmer vessels and booms collected the product. In spite of all contingency actions, some of the oil reached a coastal lagoon. The Environmental Monitoring Program, involving sampling a week and four months after the event, included chemical analysis for benzene, toluene, ethybenzene and xylenes (BTEX), petroleum hydrocarbons (TPH), aliphatic compounds (n-alkanes), polycyclic aromatic compounds (PAH), unresolved complex mixtures (UCM) and total organic carbon (TOC); toxicological tests for water using the Microtox system and Lytechinus variegatus embryo test, and for sediment using the Nitokra copepod test; and hydrocarbon detection in mussel tissue, related to local aquaculture. The coastal water PAH concentrations after the spill ranged from 0.14 to 1.92 μg/L in chrysene equivalents. For the lagoon water analysis, the concentrations of PAHs ranged from 1.54 to 9.66 μg/L. The total PAH concentrations in lagoon sediment ranged from 5.6 to 359.2 ng/g with perilene prevalence. The total PAH concentrations in mussel tissue samples ranged below the detection limit to 110.0 ng/g; the aliphatic hydrocarbon concentrations varied between 11.02 to 21.36 μg/g and total n-alkanes between 4.80 to 6.21 μg/g, showing no influence of petroleum hydrocarbons. The toxicity results were not conclusive: by means of acute tests, no toxicity was observed in the lagoon water. However, the chronic tests indicated toxicity in some samples including in the controls located in non-affected places.

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Mitochondrial impairment drives intestinal stem cell transition into dysfunctional Paneth cells predicting Crohn’s disease recurrence

Gut ◽

10.1136/gutjnl-2019-319514 ◽

2020 ◽

Vol 69 (11) ◽

pp. 1939-1951 ◽

Cited By ~ 9

Author(s):

Sevana Khaloian ◽

Eva Rath ◽

Nassim Hammoudi ◽

Elisabeth Gleisinger ◽

Andreas Blutke ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Mitochondrial Dysfunction ◽

Mitochondrial Function ◽

Mitochondrial Respiration ◽

Ex Vivo ◽

Paneth Cell ◽

Disease Recurrence ◽

Tissue Samples

ObjectiveReduced Paneth cell (PC) numbers are observed in inflammatory bowel diseases and impaired PC function contributes to the ileal pathogenesis of Crohn’s disease (CD). PCs reside in proximity to Lgr5+ intestinal stem cells (ISC) and mitochondria are critical for ISC-renewal and differentiation. Here, we characterise ISC and PC appearance under inflammatory conditions and describe the role of mitochondrial function for ISC niche-maintenance.DesignIleal tissue samples from patients with CD, mouse models for mitochondrial dysfunction (Hsp60Δ/ΔISC) and CD-like ileitis (TNFΔARE), and intestinal organoids were used to characterise PCs and ISCs in relation to mitochondrial function.ResultsIn patients with CD and TNFΔARE mice, inflammation correlated with reduced numbers of Lysozyme-positive granules in PCs and decreased Lgr5 expression in crypt regions. Disease-associated changes in PC and ISC appearance persisted in non-inflamed tissue regions of patients with CD and predicted the risk of disease recurrence after surgical resection. ISC-specific deletion of Hsp60 and inhibition of mitochondrial respiration linked mitochondrial function to the aberrant PC phenotype. Consistent with reduced stemness in vivo, crypts from inflamed TNFΔARE mice fail to grow into organoids ex vivo. Dichloroacetate-mediated inhibition of glycolysis, forcing cells to shift to mitochondrial respiration, improved ISC niche function and rescued the ability of TNFΔARE mice-derived crypts to form organoids.ConclusionWe provide evidence that inflammation-associated mitochondrial dysfunction in the intestinal epithelium triggers a metabolic imbalance, causing reduced stemness and acquisition of a dysfunctional PC phenotype. Blocking glycolysis might be a novel drug target to antagonise PC dysfunction in the pathogenesis of CD.

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Effect of culture system on the prevalence of parasites of the Mediterranean mussel Mytilus galloprovincialis (Lamark, 1819)

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.15484 ◽

2017 ◽

Vol 64 (2) ◽

pp. 113 ◽

Cited By ~ 5

Author(s):

D. KARAGIANNIS (Δ. ΚΑΡΑΓΙΑΝΝΗΣ) ◽

I. N. VATSOS (Ι.Ν. ΒΑΤΣΟΣ) ◽

A. THEODORIDIS (Α. ΘΕΟΔΩΡΙΔΗΣ) ◽

P. ANGELIDIS (Π. ΑΓΓΕΛΙΔΗΣ)

Keyword(s):

Culture System ◽

Mytilus Galloprovincialis ◽

Farming Systems ◽

Farming System ◽

Mussel Tissue ◽

Visceral Mass ◽

Tissue Samples ◽

Culture Systems ◽

Mediterranean Mussel ◽

The Mediterranean

For the production of the Mediterranean mussel, Mytilus galloprovincialis, two systems are used in Greece, the ‘long-line’ and the ‘on-table’ system. In the present study, effects of farming system on prevalence of infection by some parasites of Mediterranean mussels were investigated. Three mussel culture sites, located at Thermaikos gulf, with similar profiles, where both farming systems are practiced, were selected. Two samplings took place in 2008, one in July and one in October. From each site, 30 mussels per culture system were collected. From each mussel, tissue samples from the visceral mass, mantle and gills were collected and examined histologically for the presence of any parasites. Number of mussels parasitized by at least one parasitic species was substantially increased in mussels cultured with the ‘on-table’ system. In particular, prevalence of infection by Urastoma cyprinae and Marteilia spp. was substantially increased in mussels cultured with the ‘on-table’ system, while no relation between farming system and prevalence of infection by Eugymnanthea inquilina was observed. Prevalence of infection by Myticola intestinalis, Steinhausia mytilovum and Proctoeces maculatus was considerably low in both culture systems. In addition, in both culture systems, number of the mussels infected by E. inquilina and U. cyprinae was substantially increased in October, while an opposite trend was noted for Marteilia spp.

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A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues

International Journal of Molecular Sciences ◽

10.3390/ijms23010109 ◽

2021 ◽

Vol 23 (1) ◽

pp. 109

Author(s):

Jerome Kluza ◽

Victoriane Peugnet ◽

Blanche Daunou ◽

William Laine ◽

Gwenola Kervoaze ◽

...

Keyword(s):

Oxygen Consumption ◽

Mitochondrial Function ◽

Neurological Diseases ◽

Mitochondrial Respiratory Chain ◽

Tissue Samples ◽

Enzymatic Assays ◽

Extracellular Flux ◽

Tissue Organization ◽

New Strategy ◽

Set Up

Mitochondrial dysfunctions are implicated in several pathologies, such as metabolic, cardiovascular, respiratory, and neurological diseases, as well as in cancer and aging. These metabolic alterations are usually assessed in human or murine samples by mitochondrial respiratory chain enzymatic assays, by measuring the oxygen consumption of intact mitochondria isolated from tissues, or from cells obtained after physical or enzymatic disruption of the tissues. However, these methodologies do not maintain tissue multicellular organization and cell-cell interactions, known to influence mitochondrial metabolism. Here, we develop an optimal model to measure mitochondrial oxygen consumption in heart and lung tissue samples using the XF24 Extracellular Flux Analyzer (Seahorse) and discuss the advantages and limitations of this technological approach. Our results demonstrate that tissue organization, as well as mitochondrial ultrastructure and respiratory function, are preserved in heart and lung tissues freshly processed or after overnight conservation at 4 °C. Using this method, we confirmed the repeatedly reported obesity-associated mitochondrial dysfunction in the heart and extended it to the lungs. We set up and validated a new strategy to optimally assess mitochondrial function in murine tissues. As such, this method is of great potential interest for monitoring mitochondrial function in cohort samples.

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In situ microprobe quantitation of inorganic particle burden in paraffin sections of lung tissue

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100107010 ◽

1988 ◽

Vol 46 ◽

pp. 990-991

Author(s):

Jerrold L. Abraham

Keyword(s):

Lung Tissue ◽

Quantitative Information ◽

Particulate Material ◽

Paraffin Sections ◽

Tissue Samples ◽

Qualitative And Quantitative ◽

The Past ◽

Quantitative Analyses ◽

Data Result

Inorganic particulate material of diverse types is present in the ambient and occupational environment, and exposure to such materials is a well recognized cause of some lung disease. To investigate the interaction of inhaled inorganic particulates with the lung it is necessary to obtain quantitative information on the particulate burden of lung tissue in a wide variety of situations. The vast majority of diagnostic and experimental tissue samples (biopsies and autopsies) are fixed with formaldehyde solutions, dehydrated with organic solvents and embedded in paraffin wax. Over the past 16 years, I have attempted to obtain maximal analytical use of such tissue with minimal preparative steps. Unique diagnostic and research data result from both qualitative and quantitative analyses of sections. Most of the data has been related to inhaled inorganic particulates in lungs, but the basic methods are applicable to any tissues. The preparations are primarily designed for SEM use, but they are stable for storage and transport to other laboratories and several other instruments (e.g., for SIMS techniques).

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Oligodendrocytes in Developing Hameter Cerebral Cortex

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100090610 ◽

1976 ◽

Vol 34 ◽

pp. 126-127

Author(s):

MB. Tank Buschmann

Keyword(s):

Cerebral Cortex ◽

Optic Nerve ◽

Frontal Cortex ◽

Osmium Tetroxide ◽

Sodium Cacodylate Buffer ◽

Sodium Cacodylate ◽

Tissue Samples ◽

Cacodylate Buffer ◽

Layer V ◽

Adult Hamster

Development of oligodendrocytes in rat corpus callosum was described as a sequential change in cytoplasmic density which progressed from light to medium to dark (1). In rat optic nerve, changes in cytoplasmic density were not observed, but significant changes in morphology occurred just prior to and during myelination (2). In our study, the ultrastructural development of oligodendrocytes was studied in newborn, 5-, 10-, 15-, 20-day and adult frontal cortex of the golden hamster (Mesocricetus auratus).Young and adult hamster brains were perfused with paraformaldehyde-glutaraldehyde in sodium cacodylate buffer at pH 7.3 according to the method of Peters (3). Tissue samples of layer V of the frontal cortex were post-fixed in 2% osmium tetroxide, dehydrated in acetone and embedded in Epon-Araldite resin.

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Confocal laser microscopy of impregnated central nervous system tissue

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102559 ◽

1988 ◽

Vol 46 ◽

pp. 94-95

Author(s):

J.N. Turner ◽

M. Siemens ◽

D. Szarowski ◽

D.N. Collins

Keyword(s):

Electron Microscopy ◽

Central Nervous System ◽

Nervous System ◽

Three Dimensional ◽

Dimensional Structure ◽

Confocal Laser ◽

High Contrast ◽

Central Nervous System Tissue ◽

Tissue Samples ◽

Reflected Light

A classic preparation of central nervous system tissue (CNS) is the Golgi procedure popularized by Cajal. The method is partially specific as only a few cells are impregnated with silver chromate usualy after osmium post fixation. Samples are observable by light (LM) or electron microscopy (EM). However, the impregnation is often so dense that structures are masked in EM, and the osmium background may be undesirable in LM. Gold toning is used for a subtle but high contrast EM preparation, and osmium can be omitted for LM. We are investigating these preparations as part of a study to develop correlative LM and EM (particularly HVEM) methodologies in neurobiology. Confocal light microscopy is particularly useful as the impregnated cells have extensive three-dimensional structure in tissue samples from one to several hundred micrometers thick. Boyde has observed similar preparations in the tandem scanning reflected light microscope (TSRLM).

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An Electron Microscopic Study of the Acute Effects of Hypothalamic Releasing Factors on the Anterior Pituitary Gland

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100100901 ◽

1968 ◽

Vol 26 ◽

pp. 232-233

Author(s):

P.W. Coates ◽

E.A. Ashby ◽

L. Krulich ◽

A. Dhariwal ◽

S. McCann

Keyword(s):

Anterior Pituitary ◽

Microscopic Investigation ◽

Uranyl Acetate ◽

Electron Microscopic Investigation ◽

Electron Microscopic ◽

Tissue Samples ◽

Thin Sections ◽

Dense Membrane ◽

Membrane Bound ◽

Releasing Factors

The morphologic effects on somatotrophs of crude sheep hypothalamic extract prepared from stalk-median eminence were studied by electron microscopy in conjunction with concurrently run bioassays performed on the same tissue samples taken from young adult male Sherman rats.Groups were divided into uninjected controls and injected experimentals sacrificed at 5', 15', and 30' after injection. Half of each anterior pituitary was prepared for electron microscopic investigation, the other half for bioassay. Fixation using collidine buffered osmium tetroxide was followed by dehydration and embedment in Maraglas. Uranyl acetate and lead citrate were used as stains. Thin sections were examined in a Philips EM 200.Somatotrophs from uninjected controls appeared as described in the literature (Fig. 1). In addition to other components, these cells contained moderate numbers of spherical, electron-dense, membrane-bound granules approximately 350 millicrons in diameter.

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