Increase In Size Of Mitochondria Research Articles

Synaptic Aging is Associated with Mitochondrial Dysfunction, Reduced Antioxidant Contents and Increased Vulnerability to Amyloid-β Toxicity

Synaptic loss is the major neuropathological correlate of memory decline as a result of Alzheimer's Disease (AD). Synaptic failure appears to depend on the toxic actions of small and soluble amyloid-β (Aβ) oligomers. However, few studies have addressed the mechanism by which aging makes synapses more vulnerable to Aβ toxicity. In the present study we analyzed mitochondrial function and morphology and markers of oxidative stress in isolated presynaptic nerve endings from the hippocampus that were exposed to Aβ peptide at different ages. We found an age-related decline in mitochondrial activity, reduced antioxidant contents and increased oxidative stress markers in resting and depolarized synaptic terminals. Ultrastructural changes including an increase in mitochondrial size and a significant reduction of synaptic vesicles contents were also observed. In addition, synaptosomes obtained from 24 month old rats were more sensitive to Aβ toxicity. These data provide evidence of morphological and biochemical synaptic changes associated with aging that may contribute to exacerbate the damaging effects of Aβ.

Physiological response of Pichia pastoris GS115 to methanol-induced high level production of the Hepatitis B surface antigen: catabolic adaptation, stress responses, and autophagic processes

BackgroundPichia pastoris is an established eukaryotic host for the production of recombinant proteins. Most often, protein production is under the control of the strong methanol-inducible aox1 promoter. However, detailed information about the physiological alterations in P. pastoris accompanying the shift from growth on glycerol to methanol-induced protein production under industrial relevant conditions is missing. Here, we provide an analysis of the physiological response of P. pastoris GS115 to methanol-induced high-level production of the Hepatitis B virus surface antigen (HBsAg). High product titers and the retention of the protein in the endoplasmic reticulum (ER) are supposedly of major impact on the host physiology. For a more detailed understanding of the cellular response to methanol-induced HBsAg production, the time-dependent changes in the yeast proteome and ultrastructural cell morphology were analyzed during the production process.ResultsThe shift from growth on glycerol to growth and HBsAg production on methanol was accompanied by a drastic change in the yeast proteome. In particular, enzymes from the methanol dissimilation pathway started to dominate the proteome while enzymes from the methanol assimilation pathway, e.g. the transketolase DAS1, increased only moderately. The majority of methanol was metabolized via the energy generating dissimilatory pathway leading to a corresponding increase in mitochondrial size and numbers. The methanol-metabolism related generation of reactive oxygen species induced a pronounced oxidative stress response (e.g. strong increase of the peroxiredoxin PMP20). Moreover, the accumulation of HBsAg in the ER resulted in the induction of the unfolded protein response (e.g. strong increase of the ER-resident disulfide isomerase, PDI) and the ER associated degradation (ERAD) pathway (e.g. increase of two cytosolic chaperones and members of the AAA ATPase superfamily) indicating that potential degradation of HBsAg could proceed via the ERAD pathway and through the proteasome. However, the amount of HBsAg did not show any significant decline during the cultivation revealing its general protection from proteolytic degradation. During the methanol fed-batch phase, induction of vacuolar proteases (e.g. strong increase of APR1) and constitutive autophagic processes were observed. Vacuolar enclosures were mainly found around peroxisomes and not close to HBsAg deposits and, thus, were most likely provoked by peroxisomal components damaged by reactive oxygen species generated by methanol oxidation.ConclusionsIn the methanol fed-batch phase P. pastoris is exposed to dual stress; stress resulting from methanol degradation and stress resulting from the production of the recombinant protein leading to the induction of oxidative stress and unfolded protein response pathways, respectively. Finally, the modest increase of methanol assimilatory enzymes compared to the strong increase of methanol dissimilatory enzymes suggests here a potential to increase methanol incorporation into biomass/product through metabolic enhancement of the methanol assimilatory pathway.

Early muscle and brain ultrastructural changes in polymerase gamma 1‐related encephalomyopathy

Mutations affecting the mitochondrial DNA-polymerase gamma 1 (POLG1) gene have been shown to cause Alpers-Huttenlocher disease. Ultrastructural data on brain and muscle tissue are rare. We report on ultrastructural changes in brain and muscle tissue of two sisters who were compound heterozygous for the c.2243G>C and c.1879C>T POLG1 mutations. Patient 1 (16 years) presented with epilepsia partialis continua that did not respond to antiepileptic treatment. Neuroimaging showed right occipital and bithalamic changes. Light microscopy from a brain biopsy performed after 3 weeks suggested chronic encephalitis showing astro- and microgliosis as well as perivascular CD8-positive T-cells. However, immunosuppressive therapy failed to improve her condition. When her 17-year-old sister (patient 2) also developed epilepsy, an intensified search for metabolic diseases led to the diagnosis. On electron microscopy mitochondrial abnormalities mainly affecting neurons were detected in the brain biopsy of patient 1, including an increase in number and size, structural changes and globoid inclusions. In patient 2, light and electron microscopy on a muscle biopsy confirmed a mitochondrial myopathy, also revealing an increase in mitochondrial size and number, as well as globoid inclusions. Neurons may be the primary target of mitochondrial dysfunction in brains of patients with Alpers disease related to POLG1 mutations. During early disease stages, brain histopathology may be misleading, showing reactive inflammatory changes.

Abstract 3903: Dynamin-related Protein 1 Independent Apoptosis In Neurons Following Oxygen-glucose Deprivation

Background and purpose: Previous studies showed that mitochondrial (mito) fission occurs prior to apoptosis and that a key initiator is the mito fission protein dynamin-related protein 1 (Drp1). However, little information is available concerning mito dynamics in neurons due to OGD. Therefore, we investigated mito fission and mito fusion responses in cultured neurons in an in vitro model of brain ischemia. Methods: Primary rat cortical neurons were isolated from E18 Sprague Dawley fetuses. Neurons were exposed to OGD 9 days after culturing for 3 hr followed by up to 24 hr of reoxygenation. Mito dynamics were investigated by measuring the expression of: 1) mito fission [Drp1; mito fission 1 (Fis1)] and mito fusion [mitofusin-2 (Mfn2); optic atrophy-1 (OPA1)] proteins; 2) electron transport chain proteins (complex II 70 kDa subunit, complex IV subunit I, complex V alpha subunit); 3) mito DNA; and 4) apoptosis protein Bax. We also examined neuronal and mito morphology using transmission electron microscopy. The effects of the Drp1 inhibitors 15d-Prostaglandin J 2 (2.5-20 µM) (15d-PGJ 2 ) and Mdivi-1 (2.5-250 µM) were also investigated. Results: 3hr OGD followed by 3-24 hr recovery resulted in an increase in the level of mito DNA and enhanced expression of complex IV and V proteins. During this time, abnormal morphology, including enlarged and/or swollen mitochondria, was often observed. However, many mitochondria still retained normal morphology. Drp1 levels decreased before virtually disappearing by 6 hr of reoxygenation following OGD while Fis1 expression did not change. Bax expression increased with an increasing number of apoptotic cells during this time. Mfn2 decreased after OGD while OPA1 did not change significantly. 15d-PGJ 2 treatment dose-dependently increased cell death both in control and OGD-exposed neurons. Furthermore, 15d-PGJ 2 treatment surprisingly increased Drp1 both in controls and in OGD-exposed cells, and also increased the mito DNA. Additionally, 15d-PGJ 2 treatment itself increased the number of larger mitochondria in the neurons. However, Mdivi-1 did not have any effects. Conclusions: Despite a lower fission/fusion protein ratio, the increase in mitochondrial size indicates a shift to enhanced fusion in neurons following OGD. In addition, apoptosis induction seems to be Drp1 independent in this model. Additionally, 15d-PGJ 2 treatment appears to decrease survival in neurons following OGD. Thus, mito dynamics are affected by OGD and 15d-PGJ 2 treatment and play a role in cell survival/death.

Effect of melafen on mitochondrial apparatus of apical meristem in growth regulation in potato tubers

Growth stimulation in potato Solanum tuberosum L. tubers by melafen preparation caused an increase in area ofmitochondrial apparatus (increase in mitochondrial size) in apical meristem cells. Melafen stimulated mitochondrial differentiation (increase in number of condensed mitochondria enriched in cristas). Obtained data revealed an increase in activity of mitochondrial apparatus which is connected with an increase in energetic demands of cells in potato tuber apexes at melafen growth activation.

Chronic hypoxia-induced alterations in mitochondrial energy metabolism are not reversible in rat heart ventricles

Chronic hypoxia alters mitochondrial energy metabolism. In the heart, oxidative capacity of both ventricles is decreased after 3 weeks of chronic hypoxia. The aim of this study was to evaluate the reversal of these metabolic changes upon normoxia recovery. Rats were exposed to a hypobaric environment for 3 weeks and then subjected to a normoxic environment for 3 weeks (normoxia-recovery group) and compared with rats maintained in a normoxic environment (control group). Mitochondrial energy metabolism was differentially examined in both left and right ventricles. Oxidative capacity (oxygen consumption and ATP synthesis) was measured in saponin-skinned fibers. Activities of mitochondrial respiratory chain complexes and antioxidant enzymes were measured on ventricle homogenates. Morphometric analysis of mitochondria was performed on electron micrographs. In normoxia-recovery rats, oxidative capacities of right ventricles were decreased in the presence of glutamate or palmitoyl carnitine as substrates. In contrast, oxidation of palmitoyl carnitine was maintained in the left ventricle. Enzyme activities of complexes III and IV were significantly decreased in both ventricles. These functional alterations were associated with a decrease in numerical density and an increase in size of mitochondria. Finally, in the normoxia-recovery group, the antioxidant enzyme activities (catalase and glutathione peroxidase) increased. In conclusion, alterations of mitochondrial energy metabolism induced by chronic hypoxia are not totally reversible. Reactive oxygen species could be involved and should be investigated under such conditions, since they may represent a therapeutic target.

A12 Mitochondrial proteome analysis of R6/2 mouse brains

BackgroundSeveral observations indicate that mitochondrial dysfunction plays an important role in the pathogenesis of Huntington's disease (HD). HD patients lose significant body weight despite normal or increased food intake and...

High mitochondrial densities in the hearts of Antarctic icefishes are maintained by an increase in mitochondrial size rather than mitochondrial biogenesis

We investigated the molecular mechanisms regulating differences in mitochondrial volume density between heart ventricles of Antarctic notothenioids that vary in the expression of hemoglobin (Hb) and myoglobin (Mb). In mammals, peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) and nuclear respiratory factor 1 (NRF-1) stimulate mitochondrial biogenesis and maintain mitochondrial density in muscle tissues. We hypothesized that these factors would also maintain mitochondrial density in the hearts of Antarctic notothenioids. The percent cell volume occupied by mitochondria is significantly lower in hearts of the red-blooded notothenioid Notothenia coriiceps (18.18+/-0.69%) in comparison with those of the icefish Chaenocephalus aceratus (36.53+/-2.07%), which lacks both Hb and cardiac Mb. Mitochondrial densities are not different between hearts of N. coriiceps and Chionodraco rastrospinosus, which lacks Hb, but whose heart expresses Mb. Despite differences in mitochondrial volume density between hearts of N. coriiceps and C. aceratus, the levels of transcripts of the genes encoding PGC-1alpha, NRF-1 and citrate synthase, and the copy number of mitochondrial DNA do not differ. Our results indicate that the high mitochondrial densities in hearts of C. aceratus may result from an increase in organelle size. The surface-to-volume ratio of mitochondria from N. coriiceps is 1.9-fold greater than that of mitochondria from C. aceratus. In addition, the levels of PGC-1alpha correlate with mitochondrial density in muscle tissues of notothenioids possessing mitochondria of similar size and morphology. Finally, the levels of PGC-1alpha are 4.6-fold higher in the aerobic pectoral adductor muscle in comparison with the glycolytic skeletal muscle of N. coriiceps. The potential physiological significance of an increase in mitochondrial size in hearts of Antarctic icefishes is discussed.

Eliminating theAnt1Isoform Produces a Mouse with CPEO Pathology but Normal Ocular Motility

The adenine nucleotide transporter 1 gene (ANT1) encodes an inner mitochondrial membrane protein that transports ATP into the cell. Mutations within ANT1 produce a syndrome of chronic progressive external ophthalmoplegia (CPEO) in humans. Ant1 knockout (Ant1-/-) mice develop cardiomyopathy and mitochondrial myopathy of limb muscles. Because the extraocular muscles (EOM) are preferentially affected in human CPEO, the objective of this study was to determine whether Ant1-/- mice also exhibit an EOM mitochondrial myopathy. ANT isoform expression of isolated EOMs, EOM morphology and mitochondrial content, mitochondrial structure and function, ocular motility in intact mice, and contractile performance in isolated muscle preparations were examined. Ant1-/- EOMs had the typical appearance of mitochondrial myopathy, including increase in mitochondrial size, number, and oxidative phosphorylation (OXPHOS) staining. However, there were no measurable ocular motor abnormalities in intact Ant1-/- mice, and their isolated EOMs did not show evidence of increased fatigability. EOMs of wild-type mice exhibited higher levels of Ant2 mRNA compared with hindlimb muscle, which may compensate for the Ant1 loss in mutant mouse EOMs and account for the normal EOM function. The Ant1-/- mice provide a model in which to study CPEO pathology and compensatory mechanisms.

Aging of the liver: age-associated mitochondrial damage in intact hepatocytes.

Mitochondrial damage may be a major cause of cellular aging. So far, this hypothesis had only been tested using isolated mitochondria. The aim of this study was to investigate the involvement of mitochondria in aging using whole liver cells and not isolated mitochondria only. Using flow cytometry, we found that age is associated with a decrease in mitochondrial membrane potential (30%), an increase in mitochondrial size, and an increase in mitochondrial peroxide generation (23%). Intracellular peroxide levels were also increased. The number of mitochondria per cell and inner mitochondrial membrane mass did not change. Gluconeogenesis from glycerol or fructose (mitochondrial-independent) did not change with age, whereas it did from lactate (mitochondrial-dependent). The change in the rate of gluconeogenesis was not accompanied by changes in any of the following parameters: phosphoenolpyruvate carboxykinase or pyruvate carboxylase activities or mitochondrial ATP/ADP or cytosolic NADH/NAD+ ratios. This was caused by a decreased rate of malate export (to 20% of the controls) from mitochondria. The impairment of the mitochondrial malate transporter is posttranscriptional because its expression in Xenopus oocytes using polyadenylated RNA from livers of young or old animals did not change. Ketogenesis from oleate also fell in hepatocytes from old rats. Our results show, for the first time in intact cells, a correlation between age-associated impairment of cell metabolism and specific changes in mitochondrial function and morphology, supporting the hypothesis that mitochondrial damage plays a key role in aging.

Aging of the liver: Age-associated mitochondrial damage in intact hepatocytes

Mitochondrial damage may be a major cause of cellular aging. So far, this hypothesis had only been tested using isolated mitochondria. The aim of this study was to investigate the involvement of mitochondria in aging using whole liver cells and not isolated mitochondria only. Using flow cytometry, we found that age is associated with a decrease in mitochondrial membrane potential (30%), an increase in mitochondrial size, and an increase in mitochondrial peroxide generation (23%). Intracellular peroxide levels were also increased. The number of mitochondria per cell and inner mitochondrial membrane mass did not change. Gluconeogenesis from glycerol or fructose (mitochondrial- independent) did not change with age, whereas it did from lactate (mitochondrial-dependent). The change in the rate of gluconeogenesis was not accompanied by changes in any of the following parameters: phosphoenolpyruvate carboxykinase or pyruvate carboxylase activities or mitochondrial ATP/ADP or cytosolic NADH/NAD+ ratios. This was caused by a decreased rate of malate export (to 20% of the controls) from mitochondria. The impairment of the mitochondrial malate transporter is posttranscriptional because its expression in Xenopus oocytes using polyadenylated RNA from livers of young or old animals did not change. Ketogenesis from oleate also fell in hepatocytes from old rats. Our results show, for the first time in intact cells, a correlation between age-associated impairment of cell metabolism and specific changes in mitochondrial function and morphology, supporting the hypothesis that mitochondrial damage plays a key role in aging. (Hepatology 1996 Nov;24(5):1199-205)

Hyperthyroid Myopathy with Mitochondrial Paracrystalline Rectangular Inclusions

The ultrastructural features of a case of severe hyperthyroid myopathy are presented. Along with the moderate increase in mitochondrial size and number usually observed in most patients with hyperthyroid myopathy, some of the skeletal muscle mitochondria in the present case also contained paracrystalline rectangular inclusions. This finding has not been previously reported in hyperthyroid myopathy and further supports the current view that mitochondrial abnormalities play a major role in the pathogenesis of muscle dysfunction in hyperthyroid patients.

Chronic Rapid Atrial Pacing

Despite the clinical importance of atrial fibrillation (AF), the development of chronic nonvalvular AF models has been difficult. Animal models of sustained AF have been developed primarily in the short-term setting. Recently, models of chronic ventricular myopathy and fibrillation have been developed after several weeks of continuous rapid ventricular pacing. We hypothesized that chronic rapid atrial pacing would lead to atrial myopathy, yielding a reproducible model of sustained AF. Twenty-two halothane-anesthetized mongrel dogs underwent insertion of a transvenous lead at the right atrial appendage that was continuously paced at 400 beats per minute for 6 weeks. Two-dimensional echocardiography was performed in 11 dogs to assess the effects of rapid atrial pacing on atrial size. Atrial vulnerability was defined as the ability to induce sustained repetitive atrial responses during programmed electrical stimulation and was assessed by extrastimulus and burst-pacing techniques. Effective refractory period (ERP) was measured at two endocardial sites in the right atrium. Sustained AF was defined as AF > or = 15 minutes. In animals with sustained AF, 10 quadripolar epicardial electrodes were surgically attached to the right and left atria. The local atrial fibrillatory cycle length (AFCL) was measured in a 20-second window, and the mean AFCL was measured at each site. Marked biatrial enlargement was documented; after 6 weeks of continuous rapid atrial pacing, the left atrium was 7.8 +/- 1 cm2 at baseline versus 11.3 +/- 1 cm2 after pacing, and the right atrium was 4.3 +/- 0.7 cm2 at baseline versus 7.2 +/- 1.3 cm2 after pacing. An increase in atrial area of at least 40% was necessary to induce sustained AF and was strongly correlated with the inducibility of AF (r = .87). Electron microscopy of atrial tissue demonstrated structural changes that were characterized by an increase in mitochondrial size and number and by disruption of the sarcoplasmic reticulum. After 6 weeks of continuous rapid atrial pacing, sustained AF was induced in 18 dogs (82%) and nonsustained AF was induced in 2 dogs (9%). AF occurred spontaneously in 4 dogs (18%). Right atrial ERP, measured at cycle lengths of 400 and 300 milliseconds at baseline, was significantly shortened after pacing, from 150 +/- 8 to 127 +/- 10 milliseconds and from 147 +/- 11 to 123 +/- 12 milliseconds, respectively (P < .001). This finding was highly predictive of inducibility of AF (90%). Increased atrial area (40%) and ERP shortening were highly predictive for the induction of sustained AF (88%). Local epicardial ERP correlated well with local AFCL (R2 = .93). Mean AFCL was significantly shorter in the left atrium (81 +/- 8 milliseconds) compared with the right atrium 94 +/- 9 milliseconds (P < .05). An area in the posterior left atrium was consistently found to have a shorter AFCL (74 +/- 5 milliseconds). Cryoablation of this area was attempted in 11 dogs. In 9 dogs (82%; mean, 9.0 +/- 4.0; range, 5 to 14), AF was terminated and no longer induced after serial cryoablation. Sustained AF was readily inducible in most dogs (82%) after rapid atrial pacing. This model was consistently associated with biatrial myopathy and marked changes in atrial vulnerability. An area in the posterior left atrium was uniformly shown to have the shortest AFCL. The results of restoration of sinus rhythm and prevention of inducibility of AF after cryoablation of this area of the left atrium suggest that this area may be critical in the maintenance of AF in this model.

Hepatic ulstrastructure changes associated with the parr‐smolt transformation of Atlantic salmon (Salmo salar)

AbstractLiver samples of two groups of juvenile Atlantic salmon (Salmo salar) were collected fall through spring and examined by light and transmission electron microscopy. One group of fish was maintained under simulated natural photoperiod (SNP), the other under an 8 month advanced, phase‐shifted photoperiod (8MA). As indicated by changes in branchial Na+/K+ATPase activity, salinity tolerance, and morphological characteristics, fish in the SNP group underwent parr‐smolt transformation in the spring. Smoltification‐associated changes in ultrastructure were observed in liver parenchymal cells only. Hepatocytes of SNP smolts were smaller with markedly decreased glycogen content as compared to cells of SNP parr. An increase in mitochondrial size and a decrease in numbers of mitochondria per hepatocyte were also seen during smoltification. In smolt hepatocytes, the nucleus and mitochondria occupied a greater percent of the cell volume than in parr liver parenchymal cells. Rough and smooth endoplasmic reticulum, dense bodies, and Golgi profiles also appeared to occupy more of the cell volume of smaller hepatocytes seen during smoltification. No smoltification‐associated changes were seen in hepatocyte nuclear size, chromatin state, or nucleolar frequency. During the fall, Atlantic salmon in the 8MA group underwent some morphological and osmoregulatory changes associated with smoltification. Hepatic fine structure of these fish in the fall resembled that seen in SNP smolts. In spring, 8MA fish exhibited no indications of smoltification, and hepatocyte ultrastructure of these fish was similar to that of SNP parr. Biological implications of observations are discussed, particularly in light of other investigations concerning the liver and smoltification.

Mitochondrial growth and DNA synthesis occur in the absence of nuclear DNA replication in fission yeast

Cell growth and division require the doubling of cellular constituents followed by their equal distribution to the two daughter cells. Within a growing population, the ratio of mitochondrial to cellular volume is maintained, as is the number of mitochondrial genomes per cell. The mechanisms responsible for coordinating nuclear and mitochondrial DNA synthesis, and for balancing increases in cell and mitochondrial size are not well understood. In studies of the fission yeast Schizosaccharomyces pombe we quantified cellular and mitochondrial DNA content by both Southern blot analysis and flow cytometry of cells stained with a variety of DNA-binding fluorochromes, which we show are able to detect nuclear and mitochondrial DNA with different efficiencies. In the conditional cell division cycle mutant cdc10, which is unable to initiate nuclear DNA synthesis, we found that there was an increase in the mitochondrial DNA content in the absence of nuclear DNA replication. This demonstrates that mitochondrial and nuclear DNA synthesis are not obligately linked. We also show that mitochondrial DNA replication is not required for the increase in mitochondrial size that occurs as cells elongate, although this results in a decrease in the ratio of mitochondrial DNA to mitochondrial volume.

Comparison of the Effectiveness of Myocardial Preservation in Right Atrium and Left Ventricle

Ten patients underwent cardiac operations during which myocardial preservation was provided by systemic hypothermia, topical cardiac cooling, and cold blood cardioplegia. The duration of ischemia ranged from 45 to 142 minutes (mean, 84.2 ± 36.2 minutes). Two serial specimens (preischemic and ischemic) were obtained from the right atrium and the left ventricle, respectively; thus, a total of 40 biopsy specimens was obtained from these 10 patients. A combination of grading of ischemic injury and stereological morphometric measurement of mitochondria was performed to assess the effectiveness of myocardial preservation. Our findings from the mitochondrial score studies (grading of ischemic injury) were as follows. In the right atrium, the average mitochondrial score rose from 0.337 ± 0.235 in the preischemic stage to 1.969 ± 0.492 in the ischemic stage. In contrast, the average mitochondrial score for the left ventricle was only elevated from 0.380 ± 0.161 to 1.353 ± 0.396. The difference between preischemia of the right atrium and left ventricle is not statistically significant, but the difference between ischemia of these chambers is significant ( p < 0.01). Our stereological morphometric studies revealed that in the left ventricle, the average mitochondrial surface area was 0.316 ± 0.046 μm 2 in the preischemic stage and 0.347 ± 0.073 μm 2 in the ischemic stage, a 9.8% increase in mitochondrial size (not significant). In contrast, the mitochondrial surface area of the right atrium showed a mean increase of 65.8%, from 0.231 ± 0.038 μm 2 in the preischemic stage to 0.383 ± 0.057 μm 2 in the ischemic stage ( p < 0.001). These data showed that with regard to ultrastructural and morphometric studies of mitochondria, there was significantly poorer preservation of the right atrium compared with the left ventricle.

The effect of mycoplasmas and acholeplasmas of equine origin on organ cultures of chicken-embryo trachea

Chicken-embryo tracheal organ cultures were inoculated with equine strains of Mycoplasma arginini, M. equigenitalium, 2 strains of M. subdolum, Acholeplasma laidwii and 3 strains of A. oculi. All strains established and multiplied in the explant cultures, but only M. subdolum and A. oculi produced a cytopathic effect on ciliated epithelial cells, causing sloughing of cells and cilia after 6 days. There was a correlation between ciliostasis and increase in titre of both M. subdolum and A. oculi and this relationship was not observed with M. equigenitalium and A. laidlawii. All the strains of acholeplasma multiplied to some extent in organ culture media, but reached higher titres in the presence of explants. Cells infected with the M. subdolum strain showed sloughing of cilia, vacuolization, and increase in size of mitochondria, followed by disorganization of epithelium and marked destruction of subcellular organelles. Mycoplasmas were closely attached to the epithelial surface of the tracheal explant 8 days after infection.

Changes in ultrastructure and function of hypoxic V79 fibroblast cells after treatment with misonidazole.

Morphologic and enzymatic changes due to exposure to the radiosensitizing chemical, misonidazole, have been identified in V79 cells grown in a system in which oxygen tensions and culture density have been controlled. Misonidazole prevented the increase in mitochondrial size normally seen during exposure of these cells to conditions of moderate hypoxia (2 X 10(3) ppm O2). Mitochondrial size was also significantly decreased in cells from exponential cultures exposed to 1 mmol/l misonidazole. Morphologic changes to the mitochondria that varied from data reported elsewhere were also noted. Misonidazole caused a significant initial decrease in cytochrome oxidase activity after 4 hours of exposure of aerobic and moderately hypoxic cultures that did not return to normal in chronically hypoxic cells during continued exposure to the drug.

Comparison of Blood Cardioplegia to Electrolyte Cardioplegia on the Effectiveness of Preservation of Right Atrial Myocardium: Mitochondrial Morphometric Study

The right atrium differs from the left ventricle in two respects during cardioplegic arrest: a higher proportion of noncoronary collateral flow is delivered to the right atrium, and the atrium is frequently excluded from topical ice cooling because of its higher position relative to the left ventricle. These factors result in early rewarming of atrial myocardium. To the best of our knowledge, the surgical literature contains no reports on whether blood cardioplegia can provide better atrial myocardial preservation than electrolyte cardioplegia. Twenty consecutive patients who underwent cardiac operations were randomly selected to receive blood cardioplegia (Group 1) or electrolyte cardioplegia (Group 2). Hypothermia was achieved by systemic cooling and continuous topical cooling with ice slush. Stereological morphometric study of mitochondria was performed on 40 biopsy specimens taken from the right atrium prior to aortic cross-clamping (preischemia) and at the end of ischemia. In Group 1, total aortic cross-clamp time was 72.8 ± 32.5 minutes. The mean mitochondrial surface area before ischemia was 0.224 ± 0.032 μ 2 and after ischemia, 0.336 ± 0.032 μ 2, a 50.0% increase in mitochondrial size. In Group 2, total aortic cross-clamp time was 69.7 ± 30.9 minutes. The mean mitochondrial surface area before ischemia was 0.205 ± 0.025 μ 2 and after ischemia, 0.439 ± 0.111 μ 2, an average increase in mitochondrial size of 114.2%. There was no significant difference between the two groups in mitochondrial size before ischemia. However, after ischemia the mean mitochondrial surface areas were significantly different ( p < 0.05). We conclude that blood cardioplegia provided better preservation of right atrial myocardium than electrolyte cardioplegia with reference to the mitochondrial morphometric study.

The effect of starvation and subsequent feeding on the hepatocytes of Chanos chanos (Forsskal) fingerlings and fry

Excised liver sections of the milkfish, Chanos chanos, fry and fingerlings were studied by transmission electron microscopy. The hepatocytes underwent marked ultrastructural alterations in response to food deprivation of 10‐day starvation for fry and 2 months for the fingerlings. The prominent features characterizing the hepatocytes of starved fish were: a reduction of cell and nucleus size; apparent loss of nucleoli; condensation of chromatin material in fry; loss of stored glycogen; reduction of ER profiles; increase in the number of electron‐dense bodies containing large amounts of iron in fingerlings; and an increase in mitochondrial size. These changes were reversible following short periods of re‐feeding, i.e. 2 days for fry and 4 days for fingerlings, using natural food for the fry and formulated diet for the fingerlings.