Mitochondrial DNA Content in Human Omental Adipose Tissue

Background: Impairment of mitochondrial function plays an important role in obesity and the development of insulin resistance. The aim of this project was to investigate the mitochondrial DNA copy number in human omental adipose tissue with respect to obesity. Methods: The mitochondrial DNA (mtDNA) content per single adipocyte derived from abdominal omental adipose tissue was determined by quantitative RT-PCR in a group of 75 patients, consisting of obese and morbidly obese subjects, as well as non-obese controls. Additionally, basal metabolic rate and fat oxidation rate were recorded and expressed as total values or per kilogram fat mass. Results: MtDNA content is associated with obesity. Higher body mass index (BMI) resulted in a significantly elevated mtDNA count (ratio = 1.56; p = 0.0331) comparing non-obese (BMI < 30) to obese volunteers (BMI ≥ 30). The mtDNA count per cell was not correlated with age or gender. Diabetic patients showed a trend toward reduced mtDNA content. A seasonal change in mtDNA copy number could not be identified. In addition, a substudy investigating the basal metabolic rate and the fasting fat oxidation did not reveal any associations to the mtDNA count. Conclusions: The mtDNA content per cell of omental adipose tissue did not correlate with various clinical parameters but tended to be reduced in patients with diabetes, which may partly explain the impairment of mitochondrial function observed in insulin resistance. Furthermore, the mtDNA content was significantly increased in patients suffering from obesity (BMI above 30). This might reflect a compensatory response to the development of obesity, which is associated with impairment of mitochondrial functio

Contribution of Various Carbon Sources Toward Isoprene Biosynthesis in Poplar Leaves Mediated by Altered Atmospheric CO2 Concentrations

Biogenically released isoprene plays important roles in both tropospheric photochemistry and plant metabolism. We performed a 13CO2-labeling study using proton-transfer-reaction mass spectrometry (PTR-MS) to examine the kinetics of recently assimilated photosynthate into isoprene emitted from poplar (Populus × canescens) trees grown and measured at different atmospheric CO2 concentrations. This is the first study to explicitly consider the effects of altered atmospheric CO2 concentration on carbon partitioning to isoprene biosynthesis. We studied changes in the proportion of labeled carbon as a function of time in two mass fragments, M41+, which represents, in part, substrate derived from pyruvate, and M69+, which represents the whole unlabeled isoprene molecule. We observed a trend of slower 13C incorporation into isoprene carbon derived from pyruvate, consistent with the previously hypothesized origin of chloroplastic pyruvate from cytosolic phosphenolpyruvate (PEP). Trees grown under sub-ambient CO2 (190 ppmv) had rates of isoprene emission and rates of labeling of M41+ and M69+ that were nearly twice those observed in trees grown under elevated CO2 (590 ppmv). However, they also demonstrated the lowest proportion of completely labeled isoprene molecules. These results suggest that under reduced atmospheric CO2 availability, more carbon from stored/older carbon sources is involved in isoprene biosynthesis, and this carbon most likely enters the isoprene biosynthesis pathway through the pyruvate substrate. We offer direct evidence that extra-chloroplastic rather than chloroplastic carbon sources are mobilized to increase the availability of pyruvate required to up-regulate the isoprene biosynthesis pathway when trees are grown under sub-ambient CO2

Standardised exhaled breath collection for the measurement of exhaled volatile organic compounds by proton transfer reaction mass spectrometry

BACKGROUND: Exhaled breath volatile organic compound (VOC) analysis for airway disease monitoring is promising. However, contrary to nitric oxide the method for exhaled breath collection has not yet been standardized and the effects of expiratory flow and breath-hold have not been sufficiently studied. These manoeuvres may also reveal the origin of exhaled compounds. METHODS: 15 healthy volunteers (34 +/- 7 years) participated in the study. Subjects inhaled through their nose and exhaled immediately at two different flows (5 L/min and 10 L/min) into methylated polyethylene bags. In addition, the effect of a 20 s breath-hold following inhalation to total lung capacity was studied. The samples were analyzed for ethanol and acetone levels immediately using proton-transfer-reaction mass-spectrometer (PTR-MS, Logan Research, UK). RESULTS: Ethanol levels were negatively affected by expiratory flow rate (232.70 +/- 33.50 ppb vs. 202.30 +/- 27.28 ppb at 5 L/min and 10 L/min, respectively, p < 0.05), but remained unchanged following the breath hold (242.50 +/- 34.53 vs. 237.90 +/- 35.86 ppb, without and with breath hold, respectively, p = 0.11). On the contrary, acetone levels were increased following breath hold (1.50 +/- 0.18 ppm) compared to the baseline levels (1.38 +/- 0.15 ppm), but were not affected by expiratory flow (1.40 +/- 0.14 ppm vs. 1.49 +/- 0.14 ppm, 5 L/min vs. 10 L/min, respectively, p = 0.14). The diet had no significant effects on the gasses levels which showed good inter and intra session reproducibility. CONCLUSIONS: Exhalation parameters such as expiratory flow and breath-hold may affect VOC levels significantly; therefore standardisation of exhaled VOC measurements is mandatory. Our preliminary results suggest a different origin in the respiratory tract for these two gasses

Microflora, Contents of Polyamines, Biogenic Amines, and TVB-N in Bovine Offal and Game Meat for the Raw-Feeding of Adult Dogs

Microflora and contents of biogenic amines/polyamines and total volatile basic nitrogen (TVB-N) in 99 samples of bovine offal (red offal, n = 41 and other offal and mixes, n = 45) and wild game meat (n = 13) for raw meat-based diets (RMBD) for dogs were analyzed. Samples were bought in 11 local pet food shops and in one game-handling establishment in Austria (Lower Austria, Styria, and Vienna) in September and October 2022. Median contents (first and third quartiles in brackets) of cadaverine, histamine, tyramine, spermidine, and spermine were 20.7 [16.7; 28.6]; 25.4 [17.1; 47.2]; 18.9 [13.6; 38.9]; 15.2 [11.2; 21.2]; and 41.9 [50 mg/kg in 85.9% of samples, indicating the use of low-quality ingredients or inappropriate storage conditions. However, only 10.1% of samples were determined to be not compliant with a maximum amine content proposed for pet food. Median contents of the total aerobic bacteria counts (TACs), Pseudomonas, and Enterobacteriaceae were 7.4 [6.4; 8.0]; 6.5 [5.5; 7.7]; and 4.8 [3.9; 5.6] log CFU/g, respectively, with significantly lower counts in red offal RMBD (p Enterobacteriaceae numbers in red offal RMBD were comparable to those in food-grade red offal after 6 days of aerobic storage at 7 °C, i.e., temperatures higher than required for food-grade offal, but acceptable for animal by-products intended for RMBD production. In 80.8% of samples, numbers of Enterobacteriaceae exceeded the EU legal limit. From 12 of these samples, Salmonellae was able to be isolated, with counts from 0.03 MPN/g to 110 MPN/g. Salmonella enterica ser. Montevideo (n = 3), and S. enterica ser. Give and S. enterica ssp. Diarizonae (n = 2 each) were the most frequently isolated, while Listeria monocytogenes was rarely recovered (2%). Whilst exposure of humans handling such pet food can be reduced by hygiene precautions, the risk remains that dogs can acquire a feed-borne salmonellosis and shed the pathogen

Simultaneous real-time magnetic resonance imaging (MRI) and nose-space analysis by proton-transfer-reaction mass-spectrometry during eating

In order to better understand the performance of food products during the actual process of eating, it is important to develop techniques that allow exploring foods in-vivo and in real-time during consumption. Here we present first results on the simultaneous anal. of the eating process, combining two techniques. This includes real-time magnetic resonance imaging (MRI) to visualize the mastication, breakdown and swallowing and online nose-space anal. by proton-transfer-reaction mass-spectrometry (PTR-MS) to monitor the in-mouth aroma release

Mechanisms of hepatocellular toxicity associated with dronedarone -a comparison to amiodarone

Dronedarone is a new antiarrhythmic drug with an amiodarone-like benzofuran structure. Shortly after its introduction, dronedarone became implicated in causing severe liver injury. Amiodarone is a well-known mitochondrial toxicant. The aim of our study was to investigate mechanisms of hepatotoxicity of dronedarone in vitro and to compare them with amiodarone. We used isolated rat liver mitochondria, primary human hepatocytes, and the human hepatoma cell line HepG2, which were exposed acutely or up to 24h. After exposure of primary hepatocytes or HepG2 cells for 24h, dronedarone and amiodarone caused cytotoxicity and apoptosis starting at 20 and 50µM, respectively. The cellular ATP content started to decrease at 20µM for both drugs, suggesting mitochondrial toxicity. Inhibition of the respiratory chain required concentrations of ~10µM and was caused by an impairment of complexes I and II for both drugs. In parallel, mitochondrial accumulation of reactive oxygen species (ROS) was observed. In isolated rat liver mitochondria, acute treatment with dronedarone decreased the mitochondrial membrane potential, inhibited complex I, and uncoupled the respiratory chain. Furthermore, in acutely treated rat liver mitochondria and in HepG2 cells exposed for 24h, dronedarone started to inhibit mitochondrial β-oxidation at 10µM and amiodarone at 20µM. Similar to amiodarone, dronedarone is an uncoupler and an inhibitor of the mitochondrial respiratory chain and of β-oxidation both acutely and after exposure for 24h. Inhibition of mitochondrial function leads to accumulation of ROS and fatty acids, eventually leading to apoptosis and/or necrosis of hepatocytes. Mitochondrial toxicity may be an explanation for hepatotoxicity of dronedarone in vivo