Intervertebral Disc and Adipokine Leptin-Loves Me, Loves Me Not.

Leptin-the most famous adipose tissue-secreted hormone-in the human body is mostly observed in a negative connotation, as the hormone level increases with the accumulation of body fat. Nowadays, fatness is becoming another normal body shape. Fatness is burdened with numerous illnesses-including low back pain and degenerative disease of lumbar intervertebral disc (IVD). IVD degeneration and IVD inflammation are two indiscerptible phenomena. Irrespective of the underlying pathophysiological background (trauma, obesity, nutrient deficiency), the inflammation is crucial in triggering IVD degeneration. Leptin is usually depicted as a proinflammatory adipokine. Many studies aimed at explaining the role of leptin in IVD degeneration, though mostly in in vitro and on animal models, confirmed leptin's "bad reputation". However, several studies found that leptin might have protective role in IVD metabolism. This review examines the current literature on the metabolic role of different depots of adipose tissue, with focus on leptin, in pathogenesis of IVD degeneration.

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.

Expression of leukocyte adhesion-related glycosyltransferase genes in acute coronary syndrome patients.

INTRODUCTION: Acute coronary syndrome (ACS) is caused by destabilization and rupture of atherosclerotic plaque in the coronary artery via mechanisms affecting leukocyte signaling, rolling, adhesion, extravasation and inflammation-promoting factors. The majority of cellular communication takes place on the membrane surface that is covered with glycoproteins and glycolipids synthesized by glycosyltransferases. The aim of this study was to determine the mRNA expression of leukocyte adhesion-related glycosyltransferases in patients during the onset and the chronic phase of ACS and to compare the expression with matching subjects without coronary disease. SUBJECTS AND METHODS: The study included 26 ACS patients and 26 ACS-free matched-pair controls. Blood samples were collected at the time of hospital admittance and 8 days later. Expression analysis of six fucosyltransferases and six sialyltransferases was performed by a real-time polymerase chain reaction. RESULTS: At the time of admittance ACS subjects had lower expression levels of FUT4, ST6GalNac4, ST6Gal1 and GM3 synthase (p < 0.05) than the control subjects, and moreover, after 8 days down-regulation of FUT7 and ST6GalNac3 was also observed (p < 0.05). When compared to the initial gene expression, after treatment and stabilization of ACS subjects, FUT7, ST6GalNac2 and ST6GalNac3 were down-regulated, whereas ST6GalNac1 was up-regulated. Expression levels of FUT7, ST6GalNac1, ST6GalNac2 and ST6GalNac3 were predicted by several drugs and medical history. CONCLUSION: Expression of glycosyltransferase genes differs in ACS and control subjects. During the course of the ACS study we established further changes in gene expression levels. Medical history was predictive of gene expression levels while drugs were shown to modulate expression levels.

Emigration-related attitudes of the final year medical students in Croatia: a cross-sectional study at the dawn of the EU accession.

AIM: To investigate the emigration-related attitudes of final year medical students in Croatia at the dawn of the EU accession in 2013. METHODS: All final-year medical students at four Croatian medical schools (Zagreb, Rijeka, Split, and Osijek) were invited to participate in a cross-sectional survey on emigration attitudes. RESULTS: Among 260 respondents (response rate 61%), 90 students (35%) reported readiness for permanent emigration, expecting better quality of life (N=22, 31%), better health care organization (N=17, 24%), more professional challenges (N=10, 14%), or simply to get a job (N=8, 11%), while the least common expectation were greater earnings (N=7, 10%). The most common target countries were Germany (N=36, 40%), USA and Canada (N=15, 17%), and UK (N=10, 11%). In a multivariate analysis, readiness for permanent emigration was associated with an interest in undertaking a temporary training abroad (odds ratio [OR] 6.87; 95% confidence interval [CI] 2.83-16.72), while the belief that the preferred specialty could be obtained in Croatia appeared protective against emigration (OR 0.26; 95% CI 0.12-0.59). CONCLUSION: Despite shortages of health care workers in Croatia, the percentage of students with emigration propensity was rather high. Prevalent negative perception of the Croatian health care and recent Croatian accession to the EU pose a threat of losing newly graduated physicians to EU countries.

Croatia is a safe tourist destination - study of foreign citizen mortality in Splitsko-dalmatinska and Primorsko-goranska County during the period 2001-2010.

AIM: To investigate the mortality rate of foreign citizens in Croatia. METHODS: Data were collected from the Departments for Forensic Pathology in Split and Rijeka, which are the autopsy centers of the counties with approximately 35% of total foreign visitors, as well as from the Croatian Central Bureau for Statistics for the period 2001-2010. The mortality rate (number of deaths of members of each nationality per 100000 entrances ratio) and standardized mortality ratio (ratio between the observed and expected number of deaths) were calculated, and χ(2) goodness of fit test was used for statistical analyses. RESULTS: There were 447 deaths (325 men, 72.7%) of foreign citizens (mortality rate of 0.0015%). A total of 207 deaths (46.3%) were by natural causes, more often among older people, and 240 deaths (53.7%) were injury deaths, more often among younger people, mostly by drowning or traffic-related (22.2% and 18.6% of all deaths, respectively). Most represented were citizens of German, Austrian, Czech, and Italian nationality, with 115 (25.7%), 59 (13.2%), 58 (13.0%), and 52 deaths (11.6%), respectively. Mortality rate by nationality showed no significant difference (P<0.05). The standardized mortality ratio was lowest in Hungarian and Czech citizens (0.17) and highest in US citizens (0.35). CONCLUSIONS: Croatia has low foreign citizens' mortality rate and could be considered a safe tourist destination.

Do IL-1B and IL-1RN modulate chronic low back pain in patients with post-traumatic stress disorder?

The aim of this study was to analyze the association between single nucleotide polymorphism (SNP) in IL1B (rs1143 634) and IL1RN (rs2234677) with chronic low back pain (LBP) in chronic post-traumatic stress disorder (PTSD). A total of 406 war veterans from 1991-1995 war in Croatia participated in this study. They were divided into four groups, according to psychiatric interview, psychometric testing and the presence of LBP, verified by the imaging of lumbar area, into: (i) war veterans suffering from PTSD and LBP (N = 102), (ii) war veterans suffering from PTSD only (N = 108), (iii) war veterans suffering from LBP only (N = 99) and (iv) healthy controls (N =97). Each subject provided a blood sample for IL1B and IL1RN polymorphism testing. We found no association of rs1143634 in IL1-B with LBP Permutation test showed significant association of rs1143634 in IL1-RN with LBP group and presence of wild type allele A was protective in LBP group. The same SNP (rs1143634) in IL1-B was associated with the intensity of pain. No other associations were observed between these two markers and self-reported measures evaluating PTSD and pain symptoms. These results suggest the potential role of cytokine network in the pathogenesis of chronic PTSD and LBP, although the direct causative pathway remains unclear. The alteration of cytokine network on the level of the brain, spinal medulla and the spine may be responsible for modulation of pain and the occurrence of LBP

Genetic parameters of five new European Standard Set STR loci (D10S1248, D22S1045, D2S441, D1S1656, D12S391) in the population of eastern Croatia.

AIM: To establish allele frequencies and genetic parameters in eastern Croatia population and to compare them with those in other populations. The second aim was to compare the genetic profiles obtained with different forensic kits amplifying the same genetic markers. METHODS: Blood samples of 217 unrelated individuals from eastern Croatia were genotyped using AmpFlSTR NGM kit. Allele distribution and other genetic parameters were determined for 15 short tandem repeat (STR) loci, including the 5 loci recently added to the European Standard Set (ESS) of STR loci (D10S1248, D22S1045, D2S441, D1S1656, and D12S391). Ninety-six samples underwent duplicate analysis using AmpFlSTR Identifiler kit. RESULTS: Power of discrimination was highest for the two new ESS loci, D1S1656 (0.97254) and D12S391 (0.97339). Comparison of allele frequencies for 5 new ESS loci in our sample with previously published population data showed a significant difference from Maghreb population on D2S441 and from American Caucasian population on D1S1656. Comparison of allele frequencies for standard 10 STR loci with all the neighboring populations' data showed a significant difference only from Albanian population (on D2S1338, D18S51, and TH01). Discordant genotypes were observed in 5 (5.2%) samples at a single locus when amplified with both AmpFlSTR NGM and AmpFlSTR Identifiler kit. CONCLUSION: New ESS STR loci are highly polymorphic and short, and therefore very useful for the analysis of challenging forensic samples. DNA samples purposed for establishing databases should be routinely amplified in duplicate.

Dendritic morphology and spine density is not altered in motor cortex and dentate granular cells in mice lacking the ganglioside biosynthetic gene B4galnt1 - A quantitative Golgi cox study.

Gangliosides are characteristic plasma membrane constituents of vertebrate brain used as milestones of neuronal development. As neuronal morphology is a good indicator of neuronal differentiation, we analyzed how lack of the ganglioside biosynthetic gene Galgt1 whose product is critical for production of four major adult mammalian brain complex gangliosides (GM1, GD1a, GD1b and GT1b) affects neuronal maturation in vivo. To define maturation of cortical neurons in mice lacking B4galnt1 we performed a morphological analysis of Golgi-Cox impregnated pyramidal neurons in primary motor cortex and granular cells of dentate gyrus in 3, 21 and 150 days old B4galnt1-null and wild type mice. Quantitative analysis of basal dendritic tree on layer III pyramidal neurons in the motor cortex showed very immature dendritic picture in both mice at postnatal day 3. At postnatal day 21 both mice reached adult values in dendritic length, complexity and spine density. No quantitative differences were found between B4galnt1-null and wild type mice in pyramidal cells of motor cortex or granular cells of dentate gyrus at any examined age. In addition, the general structural and neuronal organization of all brain structures, qualitatively observed on Nissl and Golgi-Cox, were similar Our results demonstrate that neurons can develop normal dendritic complexity and length without presence of complex gangliosides in vivo. Therefore, behavioral differences observed in B4galnt1-null mice may be attributed to functional rather than morphological level of dendrites and spines of cortical pyramidal neurons.

Population substructure can significantly affect reliability of a DNA-led process of identification of mass fatality victims.

Aiming to evaluate the effects of population substructure on the reliability of a DNA correspondence in the process of human identification, we used the model of "in silico" constructed populations with and without substructure. Effects of population substructure were evaluated at the level of locus heterozygosity, Hardy-Weinberg equilibrium and mini-haplotype distribution. Inbreeding in a subpopulation of 100 individuals through 10 generations did not significantly alter the level of heterozygosity and Hardy-Weinberg equilibrium. However, analysis of mini-haplotype distribution revealed a significant homogenization in separated subpopulations. Average observed mini-haplotype frequency (f(o)) increased to threefold from expected values (f(e)), and the number of mini-haplotypes with f(o)/f(e) above 10 increased over sixfold, suggesting that the effects of population substructure on calculated likelihood ratios (LR) might be larger than previously estimated. In most criminal cases, this would not represent a problem, whereas for identifications in large-scale mass fatality events, population substructure might considerably increase the risk of false identification.

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.

Arterial tree asymmetry reduces cerebral pulsatility.

With each heartbeat, pressure wave (PW) propagates from aorta toward periphery. In cerebral circulation, at the level of circle of Willis (CW), four arteries and four PWs converge. Since the interference is an elemental property of the wave, PWs interfere at the level of CW. We hypothesize that the asymmetry of brain-supplying arteries (that join to form CW) creates phase difference between the four PWs that interfere at the level of CW and reduce downstream cerebral pulsatility. To best of our knowledge, the data about the sequence of PWs' arrival into the cerebral circulation is lacking. Evident imperfect bilateral symmetry of the vessels results with different path length of brain-supplying arteries, hence, PWs should arrive into the head at different times. The probabilistic calculation shows that asynchronous arrival is more probable than synchronous. The importance of PWs for the cerebral circulation is highlighted by the observation that barotrauma protection mechanisms are more influenced by the crest of PW (pulse pressure) than by the mean arterial pressure. In addition, an increased arterial pulsatility is associated with several brain pathologies. We created simple computational models of four converging arteries and found that asynchronous arrival of the PWs results with lower maximum pressure, slower rate of pressure amplification and lower downstream pulsatility. In analogy, the asynchronous arrival of the pressure waves into the cerebral circulation should decrease blood flow pulsatility and lower transmission of kinetic energy on arterial wall. We conclude that asynchronous arrival of PWs into the cerebral circulation influences cerebral hemodynamics and represents a physiological necessity.

Analysis of biomechanical properties of patellar ligament graft and quadruple hamstring tendon graft.

INTRODUCTION: Two types of transplant are commonly used in the surgical management of anterior cruciate ligament lesions: the central part of the patellar ligament and quadruple tendons of the gracilis muscle and semitendinosus muscle. AIMS: The aim of this study was to determine the biomechanical characteristics of patellar ligament transplants and transplants of the quadruple tendons of the hamstring muscles under tensile force in the laboratory, and to compare the results in each group of samples. MATERIALS AND METHODS: The study comprised 160 specimens: 40 specimens of gracilis muscle tendons, 40 of semitendinosus muscle tendons, 40 of quadruple tendons and 40 of the patellar ligament, approximately equally distributed according to sex, age (50-70 years) and the side of the body from which the specimen had been taken. RESULTS: The working curve analysis of the specimens under tensile load of a maximum force of 30N showed the least elongation (0.31%) in the quadruple tendon, followed by the gracilis muscle tendon (1.48%) and patellar ligament tendon (3.91%). CONCLUSIONS: The quadruple tendon specimen showed greater strength and higher elasticity compared with the patellar ligament specimen, which proved the starting hypothesis.

Epicardial Adipose Tissue Is Nonlinearly Related to Anthropometric Measures and Subcutaneous Adipose Tissue.

Introduction. Adipose tissue is the largest endocrine organ, composed of subcutaneous (SAT) and visceral adipose tissue (VAT), the latter being highly associated with coronary artery disease (CAD). Expansion of epicardial adipose tissue (EAT) is linked to CAD. One way of assessing the CAD risk is with low-cost anthropometric measures, although they are inaccurate and cannot discriminate between VAT and SAT. The aim of this study is to evaluate (1) the relationship between EAT thickness, SAT thickness and anthropometric measures in a cohort of patients assessed at the cardiology unit and (2) determine predictive power of anthropometric measures and EAT and SAT thickness in establishment of CAD. Methods. Anthropometric measures were obtained from 53 CAD and 42 non-CAD patients. Vascular and structural statuses were obtained with coronarography and echocardiography, as well as measurements of the EAT and SAT thickness. Results. Anthropometric measures showed moderate positive correlation with EAT and SAT thickness. Anthropometric measures and SAT follow nonlinear S curve relationship with EAT. Strong nonlinear power curve relationship was observed between EAT and SAT thinner than 10 mm. Anthropometric measures and EAT and SAT were poor predictors of CAD. Conclusion. Anthropometric measures and SAT have nonlinear relationship with EAT. EAT thickness and anthropometric measures have similar CAD predictive value.

Identification of person and quantification of human DNA recovered from mosquitoes (Culicidae).

Mosquitoes (Culicidae) are widespread insects and can be important in forensic context as a source of human DNA. In order to establish the quantity of human DNA in mosquitoes' gut after different post-feeding interval and for how long after taking a bloodmeal the human donor could be identified, 174 blood-engorged mosquitoes (subfamily Anophelinae and Culicinae) were captured, kept alive and sacrificed at 8h intervals. Human DNA was amplified using forensic PCR kits (Identifiler, MiniFiler, and Quantifiler). A full DNA profiles were obtained from all Culicinae mosquitoes (74/74) up to 48 h and profiling was successful up to 88 h after a bloodmeal. Duration of post-feeding interval had a significant negative effect on the possibility of obtaining a full profile (p<0.001), and logistic regression found that the probability of obtaining the full profile is 15.5% less for every 8h increase in post-feeding interval. Culicinae mosquitoes are a suitable source of human DNA for forensic STR kits more than three days after a bloodmeal. Human DNA recovered from mosquito can be used for matching purposes and could be useful in revealing spatial and temporal relation of events that took place at the crime scene. Therefore, mosquitoes at the crime scene, dead or alive, could be a valuable piece of forensic evidence.

Function of circle of Willis.

Nearly 400 years ago, Thomas Willis described the arterial ring at the base of the brain (the circle of Willis, CW) and recognized it as a compensatory system in the case of arterial occlusion. This theory is still accepted. We present several arguments that via negativa should discard the compensatory theory. (1) Current theory is anthropocentric; it ignores other species and their analog structures. (2) Arterial pathologies are diseases of old age, appearing after gene propagation. (3) According to the current theory, evolution has foresight. (4) Its commonness among animals indicates that it is probably a convergent evolutionary structure. (5) It was observed that communicating arteries are too small for effective blood flow, and (6) missing or hypoplastic in the majority of the population. We infer that CW, under physiologic conditions, serves as a passive pressure dissipating system; without considerable blood flow, pressure is transferred from the high to low pressure end, the latter being another arterial component of CW. Pressure gradient exists because pulse wave and blood flow arrive into the skull through different cerebral arteries asynchronously, due to arterial tree asymmetry. Therefore, CW and its communicating arteries protect cerebral artery and blood-brain barrier from hemodynamic stress.

Evaluation of the reliability of DNA typing in the process of identification of war victims in Croatia.

Aiming to estimate the frequency of various types of errors that can occur in the large-scale process of identification, we identified and compared genotypes of 911 parent-child pairs in the database of 3498 relatives of people that disappeared during the 1991/1992 war in Croatia. Genotypes of 891 pairs (97.8%) were matching, while 20 pairs did not match in one or more loci. Reanalysis of these samples revealed that out of 1822 analyzed genotypes, one genotype was completely wrong, and two genotypes had one wrong allele because of human errors. Five genotypes had a single wrong allele due to either polymerase chain reaction or electrophoresis errors. In five genotypes mutations were the cause of mismatch. Genetic inconsistencies with parentage were found in four "fathers" (4.2%) and three "mothers" (0.36%). As the majority of observed single-locus errors were caused by nonhuman errors, all databases produced with similar technology would probably have comparable level of errors.