The diabetes gene Zfp69 modulates hepatic insulin sensitivity in mice

AIMS/HYPOTHESIS: Zfp69 was previously identified by positional cloning as a candidate gene for obesity-associated diabetes. C57BL/6J and New Zealand obese (NZO) mice carry a loss-of-function mutation due to the integration of a retrotransposon. On the NZO background, the Zfp69 locus caused severe hyperglycaemia and loss of beta cells. To provide direct evidence for a causal role of Zfp69, we investigated the effects of its overexpression on both a lean [B6-Tg(Zfp69)] and an obese [NZO/B6-Tg(Zfp69)] background. METHODS: Zfp69 transgenic mice were generated by integrating the cDNA into the ROSA locus of the C57BL/6 genome and characterised. RESULTS: B6-Tg(Zfp69) mice were normoglycaemic, developed hyperinsulinaemia, and exhibited increased expression of G6pc and Pck1 and slightly reduced phospho-Akt levels in the liver. During OGTTs, glucose clearance was normal but insulin levels were significantly higher in the B6-Tg(Zfp69) than in control mice. The liver fat content and plasma triacylglycerol levels were significantly increased in B6-Tg(Zfp69) and NZO/B6-Tg(Zfp69) mice on a high-fat diet compared with controls. Liver transcriptome analysis of B6-Tg(Zfp69) mice revealed a downregulation of genes involved in glucose and lipid metabolism. Specifically, expression of Nampt, Lpin2, Map2k6, Gys2, Bnip3, Fitm2, Slc2a2, Ppargc1α and Insr was significantly decreased in the liver of B6-Tg(Zfp69) mice compared with wild-type animals. However, overexpression of Zfp69 did not induce overt diabetes with hyperglycaemia and beta cell loss. CONCLUSIONS/INTERPRETATION: Zfp69 mediates hyperlipidaemia, liver fat accumulation and mild insulin resistance. However, it does not induce type 2 diabetes, suggesting that the diabetogenic effect of the Zfp69 locus requires synergy with other as yet unidentified genes

Non-Invasive Quantification of White and Brown Adipose Tissues and Liver Fat Content by Computed Tomography in Mice

OBJECTIVES: Obesity and its distribution pattern are important factors for the prediction of the onset of diabetes in humans. Since several mouse models are suitable to study the pathophysiology of type 2 diabetes the aim was to validate a novel computed tomograph model (Aloka-Hitachi LCT-200) for the quantification of visceral, subcutaneous, brown and intrahepatic fat depots in mice. METHODS: Different lean and obese mouse models (C57BL/6, B6.V-Lep(ob), NZO) were used to determine the most adequate scanning parameters for the detection of the different fat depots. The data were compared with those obtained after preparation and weighing the fat depots. Liver fat content was determined by biochemical analysis. RESULTS: The correlations between weights of fat tissues on scale and weights determined by CT were significant for subcutaneous (r(2) = 0.995), visceral (r(2) = 0.990) and total white adipose tissue (r(2) = 0.992). Moreover, scans in the abdominal region, between lumbar vertebrae L4 to L5 correlated with whole-body fat distribution allowing experimenters to reduce scanning time and animal exposure to radiation and anesthesia. Test-retest reliability and measurements conducted by different experimenters showed a high reproducibility in the obtained results. Intrahepatic fat content estimated by CT was linearly related to biochemical analysis (r(2) = 0.915). Furthermore, brown fat mass correlated well with weighted brown fat depots (r(2) = 0.952). In addition, short-term cold-expose (4 °C, 4 hours) led to alterations in brown adipose tissue attributed to a reduction in triglyceride content that can be visualized as an increase in Hounsfield units by CT imaging. CONCLUSION: The 3D imaging of fat by CT provides reliable results in the quantification of total, visceral, subcutaneous, brown and intrahepatic fat in mice. This non-invasive method allows the conduction of longitudinal studies of obesity in mice and therefore enables experimenters to investigate the onset of complex diseases such as diabetes and obesity

RIGHT TO ABORTION IN MODERN LAW AND IN THE PRACTICE OF THE EUROPEAN COURT OF HUMAN RIGHTS

U pogledu dometa zaštite osnovnog ljudskog prava - prava na život i danas se suočavamo s brojnim nedoumicama i nedorečenostima. Jedna od tih nedoumica jeste i pitanje prava na abortus, koje je predmet brojnih polemika među pravnicima, filozofima, medicinskim radnicima, teolozima, ali i građanima. Debate koje se vode u raznim naučnim disciplinama ukazuju na kompleksnost pitanja koje je neophodno pravno regulisati na unutrašnjem i međunarodnom nivou. Iz tog razloga je nužno pratiti i izučavati presude međunarodnih tela koje su donete u vezi sa ovim pitanjem. Budući da je najrazvijeniji sistem zaštite ljudskih prava uspostavljen u okviru Evropske konvencije za ljudska prava, a ujedno i najrelevantniji za našu državu, u ovom radu autorka se bavi proučavanjem dosadašnje prakse Evropskog suda za ljudska prava u predstavkama koje su se odnosile na pitanje odnosa prava na život i prava na abortus.A basic human right - the right to life, even today faces numerous questions when it comes to its scope. One of those questions is the issue of the right to abortion, which is the subject of numerous controversies among lawyers, philosophers, medical workers, theologists, as well as among citizens in the broadest sense. Debates that exist in various scientific disciplines indicate the complexity of these issues that needs to be legally regulated at the domestic and international level. For that reason, it is necessary to follow and study the judgments of international bodies that have been passed in connection with this issue. As the most developed system of Human Rights protection has been established within the European Convention on Human Rights, and at the same time the most relevant for our country, in this paper the author studies the current practice of the European Court of Human Rights related to the right to abortion. It is evident, from the case law presented in this paper that the Court had a very delicate and difficult task to balance between diametrically opposing rights and interests of various interested parties. The Court’s judgments show a consensus only regarding the question of the existence of the right to abortion in cases where the right to life and health of women is endangered. Opponents of abortion claim that in this case, it is not the right to abortion, but the right to life of a woman and that only then an abortion is allowed and justified to be performed, as well as that it is a conclusion that can be deduced from the Court’s case law. However, the author of this paper believes that even though the practice of the court is quite neutral, it still tends more towards granting the right to safe abortion

Diabetes prevalence in NZO females depends on estrogen action on liver fat content.

In humans and rodents, risk of metabolic syndrome is sexually dimorphic, with an increased incidence in males. Additionally, the protective role of female gonadal hormones is ostensible, as prevalence of type 2 diabetes mellitus (T2DM) increases after menopause. Here, we investigated the influence of estrogen (E2) on the onset of T2DM in female New Zealand obese (NZO) mice. Diabetes prevalence (defined as blood glucose levels >16.6 mmol/l) of NZO females on high-fat diet (60 kcal% fat) in week 22 was 43%. This was markedly dependent on liver fat content in week 10, as detected by computed tomography. Only mice with a liver fat content >9% in week 10 plus glucose levels >10 mmol/l in week 9 developed hyperglycemia by week 22. In addition, at 11 wk, diacylglycerols were elevated in livers of diabetes-prone mice compared with controls. Hepatic expression profiles obtained from diabetes-prone and -resistant mice at 11 wk revealed increased abundance of two transcripts in diabetes-prone mice: Mogat1, which catalyzes the synthesis of diacylglycerols from monoacylglycerol and fatty acyl-CoA, and the fatty acid transporter Cd36. E2 treatment of diabetes-prone mice for 10 wk prevented any further increase in liver fat content and reduced diacylglycerols and the abundance of Mogat1 and Cd36, leading to a reduction of diabetes prevalence and an improved glucose tolerance compared with untreated mice. Our data indicate that early elevation of hepatic Cd36 and Mogat1 associates with increased production and accumulation of triglycerides and diacylglycerols, presumably resulting in reduced hepatic insulin sensitivity and leading to later onset of T2DM

Food Recognition and Food Waste Estimation Using Convolutional Neural Network

In this study, an evaluation of food waste generation was conducted, using images taken before and after the daily meals of people aged between 20 and 30 years in Serbia, for the period between 1 January and 31 April in 2022. A convolutional neural network (CNN) was employed for the tasks of recognizing food images before the meal and estimating the percentage of food waste according to the photographs taken. Keeping in mind the vast variates and types of food available, the image recognition and validation of food items present a generally very challenging task. Nevertheless, deep learning has recently been shown to be a very potent image recognition procedure, while CNN presents a state-of-the-art method of deep learning. The CNN technique was implemented to the food detection and food waste estimation tasks throughout the parameter optimization procedure. The images of the most frequently encountered food items were collected from the internet to create an image dataset, covering 157 food categories, which was used to evaluate recognition performance. Each category included between 50 and 200 images, while the total number of images in the database reached 23,552. The CNN model presented good prediction capabilities, showing an accuracy of 0.988 and a loss of 0.102, after the network training cycle. The average food waste per meal, in the frame of the analysis in Serbia, was 21.3%, according to the images collected for food waste evaluation

Comparison of bioimpedance spectroscopy and X-Ray micro-computed tomography for total fat volume measurement in mice

Obesity and the metabolic syndrome are two pathologies whose prevalence are in a constant increase. Evaluation of the total fat mass but also of the distribution between visceral and subcutaneous adipose tissue are important factors while assessing the pathophysiology of these two pathologies. Computed tomography (CT) and bioimpedance (BIS) are the translational methods the most frequently used in human beings as well as in rodent models in longitudinal studies on adiposity and obesity. Surprisingly, no direct comparison of micro-CT and BIS was reported yet in mice. Therefore, the present study was carried out to evaluate and compare the accuracy and the uncertainty of measurement of micro-CT and BIS in this species. The proportion of fat mass was measured with BIS, micro-CT and direct post-mortem tissue weight, and correlations between the data were established to evaluate the accuracy of the methods but also the uncertainty of BIS and micro-CT. There were significant correlations between weights of fat tissues on scale and proportion of total fat mass determined by BIS or micro-CT (r = 0.81 and 0.86 respectively) but both methods overestimated the total fat mass, especially in the smallest animals; overestimation of fat mass was amplified with BIS compared to micro-CT. In addition BIS and micro-CT were highly correlated (r = 0.94). Test-test reliability showed a greater variability of the BIS with respect to the micro-CT (coefficient of variation = 17.2 vs 5.6% respectively). Hence, as far as subtle differences between groups or changes within one group are awaited, micro-CT may appear as the most reliable method for determination of fat mass in mice. Micro-CT, unlike BIS, will also allow to qualitatively and quantitatively differentiate between subcutaneous and visceral adipose tissues, which is of major importance in studies on adiposity and its complications