Effect of different combination of maternal and postnatal diet on adipose tissue morphology in male rat offspring.

PURPOSE: Adipose tissue expansion can occur through several different ways and, under certain conditions, can be connected with chronic inflammation. TNF-α is one of the important cytokines involved in this process. Prolonged inflammation in obesity can lead to obesity-related insulin resistance and tissue dysfunction. The aim of our study was to investigate how different combination of maternal and postnatal diet affects offspring adipose tissue morphology and adipose tissue TNF-α expression. METHODS: Ten female Sprague Dawley rats, 9 weeks old, were randomly divided into two groups and fed either standard laboratory chow or food rich in saturated fatty acids during 6 weeks and then mated with the same male rat. After birth and lactation male rat offspring from both groups were divided into four subgroups depending on the diet they were fed until 22 weeks old. Samples of white adipose tissue were taken from the subcutaneous, epididymal, and perirenal fat pad. On tissue sections, histomorphometric analysis was conducted using CellProfiler program v 2.1.1, and immunohistochemical staining for TNF-α was performed. RESULTS: Greater mean surface area of subcutaneous and epididymal adipocytes was found in groups of male rat offspring with altered diet. In perirenal adipose tissue, the highest number of adipocytes was measured in the group where both mother and offspring were fed a high-fat diet. Adipocyte staining intensity for TNF-α did not differ significantly between the groups. CONCLUSIONS: Together with our previously published data, our results lead to the conclusion that alteration of postnatal diet can lead to TNF-α and adipocyte morphology changes.

Different combinations of maternal and postnatal diet are reflected in changes of hepatic parenchyma and hepatic TNF-alpha expression in male rat offspring.

Obesity is related to increased TNF-alpha production in different tissues. TNF-alpha is connected to mitochondrial dysfunction in the liver and also development of fatty infiltration of the liver. Also, postnatal change from normal to high-fat diet causes a significant increase in TNF-alpha serum levels. The aim of this research was to determine how maternal diet and switching male offspring to a different dietary regime after lactation influences rat liver. Ten female Sprague Dawley rats at nine weeks of age were randomly divided in two groups and fed either standard laboratory chow or high-fat diet during six weeks, and then mated with the same male subject. After birth and lactation male offspring from both groups were further divided into four subgroups depending on their subsequent diet. At 22 weeks of age, the animals were weighted, sacrificed and major organs were collected and weighted. Immunohistochemistry for TNF-alpha was performed on liver, and liver samples were analyzed for pathohistological changes. The group in which mothers were fed standard chow and offspring high-fat diet had the most pronounced changes: heaviest liver, poorest histopathological findings and strongest TNF-alpha immunohistochemical staining of liver parenchyma. High-fat diet during pregnancy and lactation and switching to high-fat diet postnatally affects liver weight, histological structure and TNF-alpha expression in male offspring.

Chemerin Blood Levels are Associated with MRI Measured Volumes of Abdominal Adipose Tissue Compartments and Lifestyle Choices.

Obesity is a low-grade chronic inflammatory state, in which a cytokine chemerin with its immunometabolic modulation has an important role. We aimed to study in a healthy population relationships between serum chemerin levels, lifestyle choices and magnetic resonance imaging (MRI) assessed volumes of abdominal visceral (VAT) and subcutaneous (SAT) adipose tissues, which have different cytokine expression profiles. Overall, 73 healthy participants undertook lifestyle questionnaire and underwent anthropometric measurements along with MRI scanning of abdominal SAT and VAT. Furthermore, complete blood count was determined along with chemerin and fibrinogen serum levels. Regression model for prediction of chemerin serum levels was built after controlling for sex, age and anthropometric measures. Median serum chemerin was 141 (125-195) ng/mL. Serum chemerin had a moderate positive correlation with SAT and VAT volumes. The two most important predictors of chemerin levels were MRI detected SAT and thigh circumference. Independently, chemerin levels were associated with smoking, preference of salty food, and favoring flavor/simplicity of preparation over nutritional value of the food. Serum chemerin seems to be more strongly correlated with the volume of abdominal SAT than VAT, with certain lifestyle choices influencing chemerin levels independently of abdominal fat.

High-fat diet induced changes in lumbar vertebra of the male rat offsprings.

In obesity, bone marrow adiposity increases and proinflammatory cytokines excretion activates RANK/RANKL/OPG system, which leads to increased bone resorption. The aim of this study was to analyze trabecular and cortical bone parameters in animals exposed to the high-fat diet in utero and after lactation. Skeletal organ of interest was the fifth lumbar vertebra, which is not exposed to biomechanical loading in rats. Further aims were to determine TNF-α and IL-6 serum concentrations, and the intensity of the TNF-α immunohistochemical staining in the bone marrow. Ten female Sprague Dawley rats, nine weeks old, were randomly divided in two groups and fed either standard laboratory chow or food rich in saturated fatty acids during five weeks, and then mated with genetically similar male subjects. After birth and lactation male offsprings from both groups were divided in four subgroups depending on the diet they were fed until twenty-two weeks of age. The highest cholesterol and triglyceride concentration were found in both groups of offsprings fed with high-fat diet. The lowest trabecular bone volume, lowest trabecular number and highest trabecular separation were found in offsprings fed with high-fat diet of mothers on standard laboratory chow. The same group of offsprings was also characterized by the highest intensity of TNF-α immunostaining in the bone marrow and the highest TNF-α serum concentration, which suggest that this proinflammatory cytokine has interfered with bone metabolism, possibly by stimulation of bone resorption, which led to inadequate trabecular bone development and bone modeling of the fifth lumbar vertebra.

Effects of between generations changes in nutrition type on vaginal smear and serum lipids in Sprague-Dawley rats.

OBJECTIVE: Previous studies established that between generations changes in feeding protocol can have significant impact on reproductive physiology. The aim of the study was to determinate effects of mothers' nutrition and nutrition of the offspring on the characteristics of vaginal smear and serum lipid content. METHODS: Ten female rats were randomly divided in two groups; first group fed with food containing high content of saturated fatty acids (HFD) and the second with standard laboratory chow (CD). After coupling and lactation period their offspring were further randomly divided into two subgroups fed HFD or CD forming four study groups: (a) CD-CD, (b) CD-HFD, (c) HFD-CD and (d) HFD-HFD. The dams and offspring at the age of 37 and 18 weeks, respectively, were subjected to biochemical analysis of the blood and cytological analysis of the vaginal smears. Additionally body weight was recorded and body mass index (BMI) was calculated. RESULTS: The HFD-HFD group presented with highest levels of triglycerides and the CD-HFD with the highest levels of cholesterol. Therefore, triglyceride and cholesterol levels were significantly different among the groups (p = 0.001 and p = 0.002, respectively). Vaginal cytological smears analysis showed features of irregular phase interchanges or extended estrous phase in offspring of high-fat fed dams. CONCLUSION: Maternal HFD consumption predisposes offspring to increased risk of developing metabolic abnormalities and estrous disorders.

Transcardial gradient of adiponectin, interleukin-6 and tumor necrosis factor-α in overweight coronary artery disease patients.

BACKGROUND: Obesity is associated with coronary artery disease (CAD), where epicardial adipose tissue (EAT) express proatherogenic cytokines (i.e., interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α)) and decreases production of beneficial adiponectin. Studies on endocrine role of EAT are mostly based on assessing cytokines' mRNAs, whereas cytokine blood levels might not readily correlate. In order to get better insight into the endocrine role of thickened EAT in CAD, we assessed transcardial gradient of adiponectin, IL-6 and TNF-α. METHODS: We assessed anthropometric and ultrasound measures in cohort of fifty nondiabetic subjects (21 CAD and 29 non-CAD). Blood sampled from aortic root and coronary sinus was assayed for adiponectin, IL-6 and TNF-α, using ELISA. RESULTS: Except thicker EAT in CAD subjects, anthropometric measures were similar (overweight), with higher adiponectin in coronary sinus than in aortic root (p<0.001) in both groups. CAD subjects had lower adiponectin in aortic root (p<0.001) and higher levels of TNF-α in coronary sinus than in aortic root (p=0.05). EAT thickness positively correlated with hip circumference (p=0.038) and negatively correlated with adiponectin levels (for both p<0.05). CONCLUSIONS: Transcardial gradient of adiponectin in non-CAD and CAD overweight subjects was similar, while latter had lower systemic adiponectin level and thicker EAT. EAT with thickening reaches the threshold level of near-maximal down-regulation of adiponectin and its further thickening is not associated with continued decrease of adiponectin production. In CAD patients levels of TNF-α were higher, but IL-6 were not, and these cytokines might be flush out by lymphatic route.

Extended Eversion Carotid Endarterectomy: Computation of Hemodynamics.

BACKGROUND: Stroke prevention includes surgery for significant stenosis of internal carotid artery (ICA). Consensus on a standard approach lacks and one alternative approach is eversion carotid endarterectomy (eCEA). To overcome disadvantages of eCEA, we developed extended-eversion carotid endarterectomy (exeCEA). Aiming to investigate hemodynamics after different surgical approaches, we created computational fluid dynamics (CFD) models of exeCEA and eCEA with included progressing lumen narrowing, representation of artery restenosis at the incision line. METHODS: Blood flow velocities, volume flow rates, and wall shear stress (WSS) were established in carotid arteries from models of eCEA and exeCEA with included increasing groove (1, 1.5, 2, and 2.5 mm) at the "incision line", under input pressure of 120 and 150 mm Hg. RESULTS: For the corresponding restenosis grade, models of exeCEA had a larger orifice toward ICA, lower blood flow velocities and higher volume flow rates in ICA, with lower volume flow rates in external carotid artery. WSS values in ICA of exeCEA models were lower than in eCEA models, later reaching the thrombotic range. CONCLUSIONS: CFD showed better hemodynamic properties in exeCEA models, indicating presented approach might be better at preserving brain perfusion.

Proton pump (H+/K+-ATPase) expression in human larynx.

OBJECTIVE: The goal of our study was to investigate and to identify the existence of proton pump in different parts of larynx. The presence of acidic content in this area is known to be connected to several laryngeal diseases. It is mostly developed by upward recurrence of acidic gastric content, but there are some signs that the acid can be produced in the larynx as well, because of the proton pump activity in laryngeal mucosa. METHODS: The study was performed on two types of specimens: (1) 50 cadaver larynges and (2) 11 surgical larynges obtained after laryngectomy. Samples were taken from supraglottis, glottis and subglottic areas and immunohistochemistry for the beta subunit of the proton pump was done. RESULTS: The presence of proton pump was proved in seromucous glands in laryngeal supraglottic area, but it was also, for the first time, found in human chondrocytes in the thyroid and epiglottic cartilage. CONCLUSION: These new findings could encourage further research that would illuminate better the etiopathogenesis not only of laryngopharyngeal reflux, but also the pathophysiology of cartilaginous disorders.

Advantages of modified osteosynthesis in treatment of osteoporotic long bones fractures--experimental model.

In surgery of fractured long bones, a patient suffering from osteoporosis represents constant challenge to a surgeon and applied material and instruments that need to destroy as little as possible of an already damaged bone. One potential way of increasing the contact surface between the implants and osteoporotic bone is injection of bone cement (methyl-metacrilat, Palakos) into a prepared screw bed. This method of osteosynthesis was therefore subjected to experimental research to prove that application of modified osteosynthesis using bone cement in treatment of fractures in osteoporotic patients has advantage over the standard method of osteosynthesis because this modified method enables significantly greater firmness and stability of the osteosynthesis, which is the essential precondition of a successful fracture healing. The research was carried out on six macerated cadaveric preparations of a shin bone from the osteological collection from Institute for Anatomy, School of Medicine, University "J. J. Strossmayer". All samples of long bones were artificially broken in the middle part of the diaphysis and then standard osteosynthesis and modified osteosynthesis with screws filled with bone cement were performed on the samples. Results show that under identical static action of the moment of torsion in the modified osteosynthesis torsion angle deviation is lower than in the standard osteosynthesis. In modified osteosynthesis with bone cement the first results for angle of torsion deviation greater than 0.2 degrees were noticed after 120 minutes, while in the standard method of osteosynthesis they were noticed already in the first minute.

Morphometric analysis of the sternum.

Sternum has a great clinical significance, considering that median sternotomy is the most common surgical approach used in cardiac surgery. The aim of this study is to standardize the sternum according to size, shape and sex and to obtain ranges of the "standard sternum". The study was done on 55 male and 35 female sterna of the average age of 65. Complex morphometric analysis of breadth, length and thickness of the sterna were performed on sternal segments which were defined by costal notches. Morphometric analysis shows that the general sternum structure in the females and in the males is equal. The standard dimensions of female and male sternum were determined. Standardization according to shape suggests that there is one standard sternum shape present in more than 2/3 of analysed samples of both sexes.

Resorption of gas trapped in body cavities: comparison of alveolar and pleural space with inner ear and paranasal sinuses.

This paper describes our attempt to devise a short text aimed at improving students' understanding of gas resorption in body cavities. Students are expected to understand the mechanisms behind paranasal sinusitis, otitis media, closed pneumothorax, and atelectasis of collapsed lung tissue, all used as examples. On the basis of the interpretation that during pneumothorax resorption, gas diffuses down pressure gradients into the blood, students are encouraged to calculate tables of pressure gradients for the above-mentioned pathological conditions. After answering a few questions, students need to analyze and eventually accept the following conclusion: in cases of air trapping in collapsible body cavities, all gases will be fully reabsorbed without pain. Air trapping in bone cavities leads only to partial reabsorption of gases and results in subatmospheric intracavity pressure. Partial vacuum causes painful mucosal edema and free fluid secretion.

Atmospheric pressure as a force that fills developing bones with marrow and air.

Many theories try to explain the existence and function of paranasal sinuses. This paper is an attempt to correlate process of paranasal sinus development in human with bone pneumatization processes in animals. It is here proposed that this mechanism starts in utero and continues after birth. During endochondral development, a solid hyaline cartilage model transforms into long bones. Central chondrocytes hypertrophy and their lacunae become confluent. Dissolving of the cartilage intercellular matrix forms a primitive marrow cavity. It is soon invaded by the periostal bud. Once circulation is established in the developing bone, the dissolved hyaline matrix can be slowly washed away from the bone cavity. Circulation in the bone cavity can develop slight subatmospheric pressures, similar to negative interstitial pressures in subcutaneous tissues. The amniotic fluid conducts atmospheric pressure to the fetal body. The pressure is trying to fill enlarging bone cavities through the existing vascular openings, or to create new openings. Bone walls of developing paranasal bones are to weak to resist the pressure gradient on their walls. New openings form on the weakest spots allowing airway mucosa to form initial paranasal sinuses. The enlarging cavities of long bones that are remote from the body surface and airway also develop a slightly subatmospheric pressure that fills them with cellular elements. These elements enter bone through the feeding vessels and form bone marrow. During after birth skeletal growth, bone remodeling shapes paranasal sinuses in a process of slow evolution that do not require measurable pressure gradients. When two sinuses come in vicinity, their growth rate declines, since the remaining thin and fragile bone lamella between them does not retract anymore.