Publications by authors named "Ivica Grkovic"

40 Publications

Expression of Leukocytes Following Myocardial Infarction in Rats is Modulated by Moderate White Wine Consumption.

Nutrients 2019 Aug 14;11(8). Epub 2019 Aug 14.

Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Šoltanska 2, Split, 21000, Croatia.

How moderate white wine consumption modulates inflammatory cells infiltration of the ischemic myocardium following permanent coronary ligation was the key question addressed in this study. Male Sprague-Dawley rats were given either a combination of different white wines or water only for 28 days. Three peri-infarct/border zones and a control/nonischemic zone were analysed to determine the expression of myeloperoxidase (MPO) and cluster of differentiation 68 (CD68). Smaller expressions for both MPO and CD68 were found in all three peri-infarct zones of wine drinking animals ( < 0.001). There was no difference in the expression of leukocyte markers between animals drinking standard and polyphenol-rich white wine, although for CD68, a nonsignificant attenuation was noticed. In sham animals, a subepicardial MPO/CD68 immunoreactive "inflammatory ring" is described. Standard white wine consumption caused attenuation of the expression of MPO but not of CD68 in these animals. We conclude that white wine consumption positively modulates peri-infarct inflammatory infiltration.
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http://dx.doi.org/10.3390/nu11081890DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722553PMC
August 2019

Disturbed Fatty Acid Oxidation, Endoplasmic Reticulum Stress, and Apoptosis in Left Ventricle of Patients With Type 2 Diabetes.

Diabetes 2019 10 7;68(10):1924-1933. Epub 2019 Aug 7.

Department of Physiology, University of Split School of Medicine, Split, Croatia

Chronic heart failure is a common complication in patients with type 2 diabetes mellitus (T2DM). T2DM is associated with disturbed metabolism of fat, which can result in excessive accumulation of lipids in cardiac muscle. In the current study, we assessed mitochondrial oxidation of carbohydrates and fatty acids, lipid accumulation, endoplasmic reticulum (ER) stress, and apoptosis in diabetic left ventricle. Left ventricular myocardium from 37 patients (a group of patients with diabetes and a group of patients without diabetes [ejection fraction >50%]) undergoing coronary artery bypass graft surgery was obtained by subepicardial needle biopsy. The group with diabetes had a significantly decreased rate of mitochondrial respiration fueled by palmitoyl-carnitine that correlated with blood glucose dysregulation, while there was no difference in oxidation of pyruvate. Diabetic myocardium also had significantly decreased activity of hydroxyacyl-CoA dehydrogenase (HADHA) and accumulated more lipid droplets and ceramide. Also, markers of ER stress response (GRP78 and CHOP) and apoptosis (cleaved caspase-3) were elevated in diabetic myocardium. These results show that, even in the absence of contractile failure, diabetic heart exhibits a decreased mitochondrial capacity for β-oxidation, increased accumulation of intracellular lipids, ER stress, and greater degree of apoptosis. Lower efficiency of mitochondrial fatty acid oxidation may represent a potential target in combating negative effects of diabetes on the heart.
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http://dx.doi.org/10.2337/db19-0423DOI Listing
October 2019

The Medical School of the Catholic University of Croatia: Principles, Goals, Standards and Organization.

Acta Med Acad 2018 May;47(1):61-75

Catholic University of Croatia, Zagreb, Croatia.

The aim of the study was to present the concept on which the School of Medicine at the Catholic University of Croatia (CUC) will be established. The new School will alleviate the shortage of physicians in Croatia and introduce an innovative form of medical education focused on principles of patient-centered care and social accountability. At the same time, the students will acquire all relevant competencies and levels of knowledge, skills and attitudes that are required by current evidence in medical education, European standards and guidelines for quality assurance at higher education institutions. The four pillars of the CUC Medical School are: 1) distributed medical education that involves health institutions outside major medical centers, 2) the concept of transformative learning, 3) teaching and practicing evidence-based medicine, and 4) implementation of quality management principles supported by information technology solutions for effective management of learning, research and practice. The overall aim of the CUC School of Medicine is to educate and train physicians capable of using best available medical evidence to deliver economically sustainable healthcare that can improve equity and health outcomes in the communities they serve, particularly those that are currently underserved.

Conclusion: The proposed programme is introducing an original system of modern medical education that insists on developing humanistic aspects of medicine, patient-centred care and social accountability, while maintaining all competencies and knowledge levels that a physician should have according to the current understanding of medical education.
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http://dx.doi.org/10.5644/ama2006-124.215DOI Listing
May 2018

Expression of adhesion molecules on granulocytes and monocytes following myocardial infarction in rats drinking white wine.

PLoS One 2018 10;13(5):e0196842. Epub 2018 May 10.

Department of Pharmacology, University of Split School of Medicine, Split, Croatia.

Neutrophils and monocytes through their CD15s, CD11b and CD44 adhesion molecules are implicated in the initiation and resolution of cardiac inflammation as well as in healing processes after the myocardial infarction (MI). The aim of this study was to determine the effect of white wine consumption on granulocyte and monocyte CD15s, CD11b, and CD44 expression 24h after the surgically inflicted MI. Granulocytes and monocytes were analyzed by flow cytometry, using whole blood of male Sprague-Dawley rats that consumed white wine for 4 weeks. This group was compared with water only drinking controls, sham animals (subject to surgery without myocardial infarction) and baseline group (intact animals that received no intervention prior to being sacrificed). Sham animals did not differ from baseline animals in CD11b+CD44+ percentage and CD44+ median fluorescence intensity. Wine drinking was associated with striking increase in CD44 expression on monocyte subpopulations. Its expression was three and fourfold increased on monocytes and large monocytes, respectively, relative to the water only drinking controls. Because of known role of CD44 on suppression of post-infarction inflammation, its upregulation on granulocytes and monocytes may significantly contribute to the microenvironment favourable for the cardiac regeneration.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0196842PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5945017PMC
August 2018

Effects of White Wine Consumption on Weight in Rats: Do Polyphenols Matter?

Oxid Med Cell Longev 2017 31;2017:8315803. Epub 2017 Oct 31.

Department of Pharmacology, University of Split School of Medicine, Split, Croatia.

Introduction: Effects of white wine and the role of wine polyphenols on weight gain in rats of different age were examined in the 4-week-voluntary-consumption trial.

Methods And Materials: Biochemically characterized standard (low polyphenols, W) and macerated (high polyphenolic content, PW) white wines were compared. One- and three-month-old Sprague-Dawley male rats ( = 78) were used. Each age group was subdivided into water-only-drinking controls (C), W, and PW-drinking animals. Daily wine and total liquid consumption, food intake, and body weight were measured, and energy intake and feed efficiency index were calculated.

Results: In both age categories, wine-drinking animals consumed less food and gained less weight in comparison to C (181 ± 2, 179 ± 6, and 201 ± 5 in younger animals and 32 ± 5, 28 ± 6, and 47 ± 4 grams in older animals, resp.), regardless of wine type. Total energy intake was the lowest in PW-drinking animals.

Conclusion: Wine-drinking animals gained less weight in comparison to C, regardless of the wines' polyphenol content. Although our results are indicative of the major role of nonphenolic constituents of the wines (probably ethanol), the modifying role of wine phenolics on weight gain cannot be excluded as the group consuming PW had lower total energy intake than other groups.
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http://dx.doi.org/10.1155/2017/8315803DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5684572PMC
July 2018

White Wine Consumption Influences Inflammatory Phase of Repair After Myocardial Infarction in Rats.

J Cardiovasc Pharmacol 2017 Nov;70(5):293-299

Departments of *Anatomy, Histology and Embryology; and †Pharmacology, University of Split School of Medicine, Split, Croatia.

Effects of white wine (WW) consumption on the expression of inflammatory markers/mediators (MMP-2, MMP-9, NF-ĸB p65 and TGF-β1) in myocardial tissue after experimentally induced permanent myocardial ischemia was investigated. Male Sprague-Dawley rats were given either a combination of WW and water or only water, for 28 days. After coronary ligation, animals were left to survive for 24 hours. Three representative areas: infarct/ischemic, peri-infarct/border zone, and control/non-ischemic zones were analyzed for expression of immunoreactivity by measuring the threshold area % of signal density. For MMP-9, significantly smaller expression was found in all 3 zones of wine drinking animals (P < 0.001). There was no difference in MMP-2 immunoreactivity between the 2 groups, except in peri-infarct zones, where the signal was significantly decreased (P < 0.001). The same pattern of expression was found for the NF-κB p65 signal, although no differences between experimental groups were observed for TGF-β1. White wine consumption decreases the expression of the 3 investigated inflammatory markers/mediators in the peri-infarct zone, suggesting its significant modulatory effect. For MMP-9 and MMP-2, expression was similar to the effect of postischemic reperfusion. No effect on TGF-β1 was observed, highlighting its role in being the master-switch, changing from the inflammatory to the proliferative stage of infarct healing.
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http://dx.doi.org/10.1097/FJC.0000000000000519DOI Listing
November 2017

Management of teaching processes using the Share point platform: A case study from the University of Split School of Medicine.

Acta Med Acad 2016 May;45(1):34-8

University of Split School of Medicine Croatia, Split, Croatia.

Objective: To develop a software tool that will combine teaching timetables with the generation of reports on teaching load.

Methods: The University of Split School of Medicine project team and the external experts from the company LAMA, LLC. determined necessary functionalities and developed the software platform as an extension of the existing software solutions already in use by the Croatian academic community.

Results: By combining comprehensive scheduling functionality with planned and performed teaching activities we determined the teaching load and realized automatic generation of payments for adjunct lecturers. The implementation required perfecting of the human resources services, brought about a manifold alleviation of the work of the entire school's administration and substantially increased the effectiveness of the quality management. The software is currently managing 54,676 teaching hours, 841 teaching staff member, 111 teaching rooms, 8 study programs, and 645 courses.

Conclusion: The program resolved several administrative problems of the school and is an example of successful implementation of IT technology in medical school management.
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http://dx.doi.org/10.5644/ama2006-124.154DOI Listing
May 2016

The business process management software for successful quality management and organization: A case study from the University of Split School of Medicine.

Acta Med Acad 2016 May;45(1):26-33

University of Split School of Medicine, Croatia.

Objective: Our aim was to describe a comprehensive model of internal quality management (QM) at a medical school founded on the business process analysis (BPA) software tool.

Methods: BPA software tool was used as the core element for description of all working processes in our medical school, and subsequently the system served as the comprehensive model of internal QM.

Results: The quality management system at the University of Split School of Medicine included the documentation and analysis of all business processes within the School. The analysis revealed 80 weak points related to one or several business processes.

Conclusion: A precise analysis of medical school business processes allows identification of unfinished, unclear and inadequate points in these processes, and subsequently the respective improvements and increase of the QM level and ultimately a rationalization of the institution's work. Our approach offers a potential reference model for development of common QM framework allowing a continuous quality control, i.e. the adjustments and adaptation to contemporary educational needs of medical students.
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http://dx.doi.org/10.5644/ama2006-124.153DOI Listing
May 2016

Expression pattern of CYP24 in liver during ageing in long-term diabetes.

Acta Histochem 2016 Jun 9;118(5):486-95. Epub 2016 May 9.

Laboratory for Neurocardiology, Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Croatia; Laboratory for Early Human Development, Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Croatia. Electronic address:

Association of liver calcitriol (active vitamin D metabolite) catabolism with osteomalacia during prolonged use of certain drugs was reported in several recent studies. To examine whether the increased calcitriol catabolism could be a potential link between ageing/diabetes mellitus (DM) and bone loss, we studied the dynamic of expression of CYP24, the main calcitriol catabolising enzyme in the liver of rats during ageing and a long-term experimental DM1. DM1 model was induced with intraperitoneally injected streptozotocin (STZ) (55mg/kg). Sprague-Dawley rats were sacrificed 6 and 12 months after the DM1 induction. The immunohistochemical analyses of CYP24 and transforming growth factor β 1 (TGF-β1) expression in the liver were performed. We found that ageing and long-term DM1 resulted in a significantly increased expression of CYP24 in hepatocytes, as well as in non-hepatocyte liver cells (Kupffer cells, hepatic stellate cells and sinusoidal endothelial cells). Ageing and long-term DM1 resulted in an increased expression of TGF-β1 as well. Expression of CYP24 coexisted with the expression of TGF-β1 in all types of hepatic cells. We concluded that liver has the capacity for an active vitamin D catabolism in different populations of liver cells, especially in sinusoidal endothelial cells, through an expression of CYP24. That capacity is substantially increased during ageing and long-term diabetes mellitus. Increased liver calcitriol catabolism could be one of the mechanisms of the bone metabolism impairment related to ageing and diabetes.
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http://dx.doi.org/10.1016/j.acthis.2016.05.001DOI Listing
June 2016

Aging and a long-term diabetes mellitus increase expression of 1 α-hydroxylase and vitamin D receptors in the rat liver.

Exp Gerontol 2015 Dec 22;72:167-76. Epub 2015 Oct 22.

Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Šoltanska 2, 21000 Split, Croatia. Electronic address:

Diabetes mellitus (DM) is a metabolic disorder associated with serious liver complications. As a metabolic chronic disease, DM is very common in the elderly. Recent studies suggest ameliorating effects of vitamin D on metabolic and oxidative stress in the liver tissue in an experimental model of DM. The aim of this study was to investigate the expression of vitamin D receptors (VDRs) and 1α-hydroxylase, the key enzyme for the production of active vitamin D form (calcitriol) in the liver during long-term diabetes mellitus type 1 (DM1) in aging rats. We performed immunohistochemical analysis of liver expression of 1α-hydroxylase and VDRs during aging in long-term streptozotocin-induced DM1. 1α-Hydroxylase was identified in the monocyte/macrophage system of the liver. In addition to the nuclear expression, we also observed the expression of VDR in membranes of lipid droplets within hepatocytes. Aging and long-term DM1 resulted in significant increases in the number of 1α-hydroxylase immunoreactive cells, as well as the percentage of strongly positive VDR hepatocytes. In conclusion, the liver has the capacity for active vitamin D synthesis in its monocyte/macrophage system that is substantially increased in aging and long-term diabetes mellitus. These conditions are also characterized by significant increases in vitamin D receptor expression in hepatocytes. The present study suggests that VDR signaling system could be a potential target in prevention of liver complications caused by diabetes and aging.
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http://dx.doi.org/10.1016/j.exger.2015.10.005DOI Listing
December 2015

Diabetes mellitus affects activity of calcium/calmodulin-dependent protein kinase II alpha in rat trigeminal ganglia.

J Chem Neuroanat 2015 Mar-Apr;64-65:12-9. Epub 2015 Feb 17.

Department of Anatomy, Histology and Embryology, University of Split, School of Medicine, Šoltanska 2, 21000 Split, Croatia. Electronic address:

The activity of calcium/calmodulin-dependent protein kinase II alpha (CaMKIIα) may play a critical role in the modulation of nociceptor activity and plasticity of primary sensory trigeminal neurons. The aim of this study was to investigate the immunoreactivity of phosphorylated CaMKIIα (pCaMKIIα) in subpopulations of trigeminal ganglion (TG) neurons in rat models of early diabetes type 1 (dm1) and 2 (dm2). DM1 model was induced with intraperitoneally (i.p.) injected streptozotocin (STZ) (55mg/kg). DM2 rats were fed with the high fat diet (HFD) for 2 weeks and then received 35mg/kg of STZ i.p. Two weeks and 2 months after the STZ-diabetes induction, rats were sacrificed and immunohistochemical analysis for detection of pCaMKIIα immunoreactivity and double immunofluorescence labelling with isolectin (IB4) was performed. Increased intensity of pCaMKIIα immunofluorescence, restricted to IB4-negative small-diameter neurons, was seen in TG neurons two months after STZ-DM1 induction. DM1 model, as well as the obesity (control dm2 groups) resulted in neuronal impaired growth while dm2 model led to neuron hypertrophy in TG. Observed changes may play a critical role in the modulation of nociceptor activity and plasticity of primary sensory trigeminal neurons. In future, innovative strategies for modulation of CaMKIIα activity in specific subpopulations of neurons could be a novel approach in therapy of diabetic trigeminal neuropathy.
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http://dx.doi.org/10.1016/j.jchemneu.2015.02.003DOI Listing
February 2016

Expression of nestin in superior cervical ganglia of rats is influenced by gender and gonadectomy.

J Chem Neuroanat 2015 Jan 4;63:6-12. Epub 2014 Dec 4.

Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Šoltanska 2, 21000 Split, Croatia.

Neurons and glia arise from neural progenitor cells that express nestin. Although substantial changes in neuronal development were observed during the postnatal period, data concerning dynamics of nestin expression in the superior cervical ganglia (SCG) of rat during that period are lacking. It is known that gonadectomy and steroid hormones influence the development of neurons in the SCG during the postnatal period, but there are no data on how they influence the persistence of nestin expression in the SCG cells. The dynamics of nestin expression in the SCG in rats of three age groups, as well as the influence of gender and gonadectomy, was investigated. Three groups of male rats were sacrificed at 2, 3 and 6 months of age. Additional groups of male and female Sprague-Dawley rats were gonadectomized at the age of 2 months. After 30 days, they were sacrificed and SCGs were harvested and processed immunohistochemically. Immunoreactivity for nestin in the SCG was observed in satellite glia, based on their expression of s100. The proportion of neurons that were encircled with nestin-immunoreactive satellite cells (nestin encircled neurons, NEN) decreased between second and third month of age (p<0.05). The proportion of NEN was greater in the NPY+ than in the NPY- subpopulation. The proportion of NEN in the SCG of female rats was significantly higher (p<0.05) than that of both, the male rats and ovariectomised groups. The percentage of these neurons was significantly higher (p<0.05) in orchidectomised, in comparison to male rats. Results show the existence of nestin-immunoreactive satellite cells in the SCG of adult rats. A substantial decrease of nestin expression in SCG cells of rats, after the onset of sexual maturation, was observed. This decrease showed significant sex-dependence and was dramatically influenced by gonadal activity.
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http://dx.doi.org/10.1016/j.jchemneu.2014.11.003DOI Listing
January 2015

Effects of isoflurane postconditioning on chronic phase of ischemia-reperfusion heart injury in rats.

Cardiovasc Pathol 2015 Mar-Apr;24(2):94-101. Epub 2014 Sep 16.

Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Split, Croatia.

Introduction: The application of isoflurane in a postconditioning manner, during early reperfusion of ischemic myocardium, reduces the infarct size. Its favorable effect on highly vascularized granulation tissue formation is very important considering the fact that increased genesis of blood vessels in peri-infarct zone reduces the infarct size and improves cardiac function. Taking into consideration the influence of isoflurane on the subacute phase of infarct healing, by using different immunohistochemical markers, we wanted to explore whether isoflurane postconditioning influences the chronic phase of healing.

Methods: The size of infarcted region was measured, and comparisons between isoflurane-treated and control animals were made. Quality of infarcted area was assessed by detecting vascular endothelial growth factor (VEGF), platelet/endothelial cell adhesion molecule-1 (PECAM-1/CD31) as a marker of angiogenesis, and nestin as a marker of immature progenitor cells, and de novo formed blood vessels (vasculogenesis).

Results: There was no difference between the control and isoflurane-treated groups in VEGF and PECAM-1/CD31 expression. However, a large reduction in infarct size was found (68.1% of control). Also, a marked decrease of nestin expression in immature progenitor cells, along with a marked increase of the same marker in cardiomyocytes, (signs of myocardium regeneration), was found in experimental animals when compared to control animals that did not receive isoflurane treatment.

Conclusions: Based on our results, we can emphasize two morphologically detectable benefits of isoflurane postconditioning: a marked reduction in infarct size along with a more mature-looking infarct area in the chronic phase of infarct healing.
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http://dx.doi.org/10.1016/j.carpath.2014.09.004DOI Listing
November 2015

Expression of PTHrP and PTH/PTHrP receptor 1 in the superior cervical ganglia of rats.

Neuropeptides 2014 Dec 20;48(6):353-9. Epub 2014 Sep 20.

Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Šoltanska 2, 21000 Split, Croatia.

PTHrP and its receptor PTHR1 are found in the CNS and peripheral nervous system. The presence of PTHrP mRNA has been detected in the superior cervical ganglion (SCG), but there are no data on the cellular distribution of PTHrP and PTHR1 in the SCG. Although it is known that ovarian activity and reproductive status influence sympathetic activity, and the PTHrP/PTHR1 system is influenced by estrogens in different tissues, it is not known whether these factors have a similar effect on expression of PTHrP and PTHR1 in the nervous system. Hence, we investigated the presence and distribution of PTHrP and PTHR1 in neurons and glia of the SCG of rats, as well as the influence of ovariectomy on their expression, by using immunohistochemistry. PTHrP and PTHR1 immunoreactivity was observed in cytoplasm as well as in nuclei of almost all neurons in the SCG. In male rats, intensity of PTHrP fluorescence was significantly higher in cytoplasm of NPY-, in comparison to NPY+ neurons (p < 0.05). In female rats, 2 months post-ovariectomy, significantly lower intensity of PTHrP fluorescence in cytoplasm of the SCG neurons was observed in comparison to sham operated animals (p < 0.05). In addition to neurons, PTHrP and PTHR1 immunoreactivity was observed in most of the glia and was not influenced by ovariectomy. Results show the expression of PTHrP and its receptor, PTHR1, in the majority of neurons and glial cells in the SCG of rats. Expression of PTHrP, but not PTHR1 in the cytoplasm of SCG neurons is influenced by ovarian activity.
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http://dx.doi.org/10.1016/j.npep.2014.09.005DOI Listing
December 2014

Behavioral changes following experimentally-induced acute myocardial infarction in rats.

Int Heart J 2014 14;55(2):169-77. Epub 2014 Mar 14.

Department of Anatomy, Embryology and Histology, University of Split School of Medicine.

Rats with experimentally-induced acute myocardial infarction (AMI) have proven to be a clinically relevant model for visceral pain. As there are no behavioral data available on rats in the postinfarction period, we aimed to identify specific pain-related behavioral changes following AMI to increase the validity of the model. AMI was induced by left coronary artery ligation and pain-related behavior was analyzed using the open field test (OFT) and elevated plus maze (EPM). Morphine was applied following AMI induction to differentiate pain-related changes from those related to nonspecific global changes in responsiveness. AMI was histologically confirmed. Hypolocomotion was consistently evident in all behavioral tests for both the infarcted group and sham group. In the OFT, both AMI and sham rats exhibited less exploratory behavior and less activity. A similar pattern of behavior was observed in EPM, where both surgical groups showed fewer entries to the open arms and spent less time in the open arms. The sham group with an intact pericardium showed the same pattern of activity as control rats. The reduction in activity and rearing observed following AMI was successfully reversed following morphine injection. This effect was abolished after naloxone application allowing us to attribute observed changes specifically to pain.This study demonstrates that pain-related behavior in the acute postinfarction period is generally characterized by reduced mobility and explorative behavior. Our results showed that cardiac ischemia as a consequence of experimentally-induced infarction is a less important source of pain behavior than manipulation of the pericardium.
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http://dx.doi.org/10.1536/ihj.13-275DOI Listing
May 2014

Gender and gonadectomy influence on neurons in superior cervical ganglia of sexually mature rats.

Neurosci Lett 2014 Mar 28;563:55-60. Epub 2014 Jan 28.

Department of Anatomy, Histology and Embriology, University of Split School of Medicine, Šoltanska 2, 21000 Split, Croatia.

Gonadal hormones have a significant influence on both the number of neurons and the density of synapses in the superior cervical ganglion (SCG) during the early postnatal period. There are no studies reporting influence of the absence of these factors in sexually mature animals, although changes in SCG-neurons of the rat were observed up to 6 months of age. Hence, we investigated whether gonadectomy of sexually mature rats influences morphological properties of neurons in the SCG of the rat and if so, would it have a specific effect on neurochemically distinct subpopulations. Male and female Sprague-Dawley rats were gonadectomized at the age of two months. After 30 days, they were sacrificed and SCGs were harvested and processed immunohistochemically. The mean diameter of NPY- neurons was greater in male rats, in comparison to all other groups (p<0.05). The number of NPY+, NPY- and total neurons per section area was significantly higher in female than in male, orchidectomized or the ovariectomized animals (p<0.05). The share of the different neuronal populations in the SCG that were encircled with calretinin-positive baskets (c.b+) or c.b.- (NPY+ or NPY-) was significantly influenced by the gender of the animals and gonadectomy, with significantly more c.b.+ in male animals (p<0.05). Results of the present study indicate that substantial changes in the SCG neurons of the rat occur after reaching sexual maturity, and are influenced by the gonadectomy.
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http://dx.doi.org/10.1016/j.neulet.2014.01.028DOI Listing
March 2014

Changes in cardiac innervation during maturation in long-term diabetes.

Exp Gerontol 2013 Dec 16;48(12):1473-8. Epub 2013 Oct 16.

Laboratory for Neurocardiology, University of Split School of Medicine, Soltanska 2, 21000 Split, Croatia.

Diabetic autonomic neuropathy being a common complication of diabetes mellitus (DM) is related to an increased risk of cardiovascular mortality. However, mechanisms underlying changes of innervation density in affected hearts remain insufficiently understood. Hence, the aim of this study was to describe quantitative changes of intra-myocardial nerve terminals in hearts of diabetic rats of various ages. Male Sprague-Dawley rats were injected with 55mg/kg streptozotocin (STZ) (DM group) or with citrate buffer (control). After 2weeks, 2months, 6months and 12months, sections of their hearts were analyzed in five areas-left ventricle, interventricular septum, right ventricle, anterior and posterior wall. Nerve fibers were visualized immunohistochemically, using antibody against a general neuronal marker, protein gene product 9.5 (PGP 9.5). Significant increase in total nerve fibers from all heart areas was observed 2weeks and 2months after diabetes induction, followed by a decrease at 6months and again increase at 12months was observed in both control and diabetic rats. Significant difference between control and diabetic rats was visible after 2weeks and 2months, with diabetic rats exhibiting significantly more nerve fibers. There were no consistent differences in quantity of nerve fibers in different areas of the heart within a particular age-related group of animals. In conclusion, cardiac innervation undergoes dynamic changes both in control and in diabetic rats, with a time-dependent significant increase in neuronal fiber density in diabetic animals. This novel information may contribute to our understanding of pathophysiological changes associated with diabetic cardiac neuropathy.
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http://dx.doi.org/10.1016/j.exger.2013.10.004DOI Listing
December 2013

Isoflurane post-conditioning stimulates the proliferative phase of myocardial recovery in an ischemia-reperfusion model of heart injury in rats.

Histol Histopathol 2014 01 12;29(1):89-99. Epub 2013 Jul 12.

Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Croatia.

The application of isoflurane in a post-conditioning manner, during early reperfusion following a period of coronary occlusion, has numerous beneficial effects on the ischemic myocardium, including reduction of infarct size. It does so by stimulating a sequence of well studied anti-apoptotic pro-survival mechanisms in a similar manner to various 'ischemic' pre-/post-conditioning approaches which achieve their cardio protective effects in both laboratory and clinical situations. Proliferation of newly formed blood vessels, resulting in formation of highly vascularized granulation tissue, is an essential stage of infarct healing. It can be evaluated by detecting various angiogenic factors, including vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) or by quantification of expression of vascular smooth muscle progenitors, such as Nestin. Expression of these three markers was used to evaluate the effect of early isoflurane post-conditioning in ischemia-reperfusion type cardiac injury. A large reduction in infarct size (59.3% of control), and marked increase of expression of VEGF (43.4%), PECAM-1/CD31 (136%) and Nestin (77.9%) was found in experimental animals when compared to control animals that did not receive isoflurane treatment. Hence, based on our results, we can emphasize two morphologically detectable benefits of isoflurane post-conditioning: a marked reduction in infarct size and much better organization/vascularization of necrotic tissue.
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http://dx.doi.org/10.14670/HH-29.89DOI Listing
January 2014

Increased vitamin D receptor expression in dorsal root ganglia neurons of diabetic rats.

Neurosci Lett 2013 Aug 16;549:140-5. Epub 2013 May 16.

School of Medicine in Split, Department of Anatomy, Histology and Embriology, Šoltanska 2, 21000 Split, Croatia.

The effects of vitamin D on the nervous system have been studied extensively. In spite of accumulating data about the substantial changes in the vitamin D receptor (VDR) signaling system, during different types of neuroinflammatory diseases, its role in diabetic neuropathy has not been investigated in detail. To assess the role of VDR signaling in diabetic neuropathy, we examined expression of VDRs in dorsal root ganglia (DRG) neurons in a rat model of streptozotocin-induced diabetes mellitus type 1. Diabetes mellitus (DM) type 1 was induced with streptozotocin in male Sprague-Dawley rats. After two months, expression of VDRs was analyzed immunohistochemically in the cytoplasm of L4 and L5 DRG neurons of diabetic rats. Semi-quantitative analysis for the determination of staining in nuclei and plasma-membranes of DRG neurons was performed. A significant increase in VDR expression was observed in DRG neurons of diabetic rats. Expression of VDRs was increased in the cytoplasm, nuclei and in cell membranes of neurons. An increase in VDR expression occurred in all neurons, but the greatest increase of fluorescence intensity in cytoplasm was observed in neurons of small diameter. Results of the present study indicate that the VDR signaling system could be a potential therapeutic target for diabetic neuropathy.
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http://dx.doi.org/10.1016/j.neulet.2013.05.023DOI Listing
August 2013

Age-related changes of neurochemically different subpopulations of cardiac spinal afferent neurons in rats.

Exp Gerontol 2013 Aug 25;48(8):774-7. Epub 2013 Apr 25.

Department of Anatomy, Histology and Embriology, School of Medicine in Split, Šoltanska 2, 21000 Split, Croatia.

This study investigated the effect of aging on cardiac spinal afferent neurons in the rat. A patch loaded with retrograde tracer Fast Blue (FB) was applied to all chambers of the rat heart. Morphological and neurochemical characteristics of labeled cardiac spinal afferent neurons were assessed in young (2 months) and old (2 years) rats using markers for likely unmyelinated (isolectin B4; IB4) and myelinated (neurofilament 200; N52) neurons. The number of cardiac spinal afferent neurons decreased in senescence to 15% of that found in young rats (1604 vs. 248). The size of neuronal soma as well as proportion of IB4+ neurons increased significantly, whereas the proportion of N52+ neurons decreased significantly in senescence. Unlike somatic spinal afferents, neurochemically different populations of cardiac spinal afferent neurons experience morphological and neurochemical changes related to aging. A major decrease in total number of cardiac spinal afferent neurons occurs in senescence. The proportion of N52+ neurons decreased in senescence, but it seems that nociceptive innervation is preserved due to increased proportion and size of IB4+ unmyelinated neurons.
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http://dx.doi.org/10.1016/j.exger.2013.04.006DOI Listing
August 2013

Ways to address the challenges of a modern medical curriculum: living academic medicine at the University of Split, School of Medicine.

Acta Med Acad 2012 ;41(1):7-17

Department of Anatomy, Histology and Embryology, University of Split, School of Medicine, Soltanska 2, Split, Croatia.

Structure of a modern medical curriculum should follow recommendations of professional bodies (like World Federation for Medical Education) in order to educate and train medical professionals equipped with problem solving/critical thinking skills entering a world of evidence-based medicine and demands of contemporary medical practices. Also, political and socio-economic realities in addition to traditional and cultural values should be taken into account in order to avoid creating an unsustainable program. Requests for curricular changes by the European Union Commission were used as a chance to shape our program into an original blend of traditional pre-clinical and clinical subjects with several vertically integrated subjects focusing on mastering clinical skills, professional attitudes, information management and critical as well as evidence-based reasoning and decision making. Reasons for introducing curricular changes in addition to detailed structure of current medical course at the University of Split School of Medicine is presented in this paper.
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http://dx.doi.org/10.5644/ama2006-124.31DOI Listing
July 2013

Genome-wide association and large-scale follow up identifies 16 new loci influencing lung function.

Authors:
María Soler Artigas Daan W Loth Louise V Wain Sina A Gharib Ma'en Obeidat Wenbo Tang Guangju Zhai Jing Hua Zhao Albert Vernon Smith Jennifer E Huffman Eva Albrecht Catherine M Jackson David M Evans Gemma Cadby Myriam Fornage Ani Manichaikul Lorna M Lopez Toby Johnson Melinda C Aldrich Thor Aspelund Inês Barroso Harry Campbell Patricia A Cassano David J Couper Gudny Eiriksdottir Nora Franceschini Melissa Garcia Christian Gieger Gauti Kjartan Gislason Ivica Grkovic Christopher J Hammond Dana B Hancock Tamara B Harris Adaikalavan Ramasamy Susan R Heckbert Markku Heliövaara Georg Homuth Pirro G Hysi Alan L James Stipan Jankovic Bonnie R Joubert Stefan Karrasch Norman Klopp Beate Koch Stephen B Kritchevsky Lenore J Launer Yongmei Liu Laura R Loehr Kurt Lohman Ruth J F Loos Thomas Lumley Khalid A Al Balushi Wei Q Ang R Graham Barr John Beilby John D Blakey Mladen Boban Vesna Boraska Jonas Brisman John R Britton Guy G Brusselle Cyrus Cooper Ivan Curjuric Santosh Dahgam Ian J Deary Shah Ebrahim Mark Eijgelsheim Clyde Francks Darya Gaysina Raquel Granell Xiangjun Gu John L Hankinson Rebecca Hardy Sarah E Harris John Henderson Amanda Henry Aroon D Hingorani Albert Hofman Patrick G Holt Jennie Hui Michael L Hunter Medea Imboden Karen A Jameson Shona M Kerr Ivana Kolcic Florian Kronenberg Jason Z Liu Jonathan Marchini Tricia McKeever Andrew D Morris Anna-Carin Olin David J Porteous Dirkje S Postma Stephen S Rich Susan M Ring Fernando Rivadeneira Thierry Rochat Avan Aihie Sayer Ian Sayers Peter D Sly George Davey Smith Akshay Sood John M Starr André G Uitterlinden Judith M Vonk S Goya Wannamethee Peter H Whincup Cisca Wijmenga O Dale Williams Andrew Wong Massimo Mangino Kristin D Marciante Wendy L McArdle Bernd Meibohm Alanna C Morrison Kari E North Ernst Omenaas Lyle J Palmer Kirsi H Pietiläinen Isabelle Pin Ozren Pola Sbreve Ek Anneli Pouta Bruce M Psaty Anna-Liisa Hartikainen Taina Rantanen Samuli Ripatti Jerome I Rotter Igor Rudan Alicja R Rudnicka Holger Schulz So-Youn Shin Tim D Spector Ida Surakka Veronique Vitart Henry Völzke Nicholas J Wareham Nicole M Warrington H-Erich Wichmann Sarah H Wild Jemma B Wilk Matthias Wjst Alan F Wright Lina Zgaga Tatijana Zemunik Craig E Pennell Fredrik Nyberg Diana Kuh John W Holloway H Marike Boezen Debbie A Lawlor Richard W Morris Nicole Probst-Hensch Jaakko Kaprio James F Wilson Caroline Hayward Mika Kähönen Joachim Heinrich Arthur W Musk Deborah L Jarvis Sven Gläser Marjo-Riitta Järvelin Bruno H Ch Stricker Paul Elliott George T O'Connor David P Strachan Stephanie J London Ian P Hall Vilmundur Gudnason Martin D Tobin

Nat Genet 2011 Sep 25;43(11):1082-90. Epub 2011 Sep 25.

Department of Health Sciences, University of Leicester, Leicester, UK.

Pulmonary function measures reflect respiratory health and are used in the diagnosis of chronic obstructive pulmonary disease. We tested genome-wide association with forced expiratory volume in 1 second and the ratio of forced expiratory volume in 1 second to forced vital capacity in 48,201 individuals of European ancestry with follow up of the top associations in up to an additional 46,411 individuals. We identified new regions showing association (combined P < 5 × 10(-8)) with pulmonary function in or near MFAP2, TGFB2, HDAC4, RARB, MECOM (also known as EVI1), SPATA9, ARMC2, NCR3, ZKSCAN3, CDC123, C10orf11, LRP1, CCDC38, MMP15, CFDP1 and KCNE2. Identification of these 16 new loci may provide insight into the molecular mechanisms regulating pulmonary function and into molecular targets for future therapy to alleviate reduced lung function.
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http://dx.doi.org/10.1038/ng.941DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3267376PMC
September 2011

A comprehensive evaluation of potential lung function associated genes in the SpiroMeta general population sample.

PLoS One 2011 20;6(5):e19382. Epub 2011 May 20.

Nottingham Respiratory Biomedical Research Unit, Division of Therapeutics and Molecular Medicine, University Hospital of Nottingham, Nottingham, United Kingdom.

Rationale: Lung function measures are heritable traits that predict population morbidity and mortality and are essential for the diagnosis of chronic obstructive pulmonary disease (COPD). Variations in many genes have been reported to affect these traits, but attempts at replication have provided conflicting results. Recently, we undertook a meta-analysis of Genome Wide Association Study (GWAS) results for lung function measures in 20,288 individuals from the general population (the SpiroMeta consortium).

Objectives: To comprehensively analyse previously reported genetic associations with lung function measures, and to investigate whether single nucleotide polymorphisms (SNPs) in these genomic regions are associated with lung function in a large population sample.

Methods: We analysed association for SNPs tagging 130 genes and 48 intergenic regions (+/-10 kb), after conducting a systematic review of the literature in the PubMed database for genetic association studies reporting lung function associations.

Results: The analysis included 16,936 genotyped and imputed SNPs. No loci showed overall significant association for FEV(1) or FEV(1)/FVC traits using a carefully defined significance threshold of 1.3×10(-5). The most significant loci associated with FEV(1) include SNPs tagging MACROD2 (P = 6.81×10(-5)), CNTN5 (P = 4.37×10(-4)), and TRPV4 (P = 1.58×10(-3)). Among ever-smokers, SERPINA1 showed the most significant association with FEV(1) (P = 8.41×10(-5)), followed by PDE4D (P = 1.22×10(-4)). The strongest association with FEV(1)/FVC ratio was observed with ABCC1 (P = 4.38×10(-4)), and ESR1 (P = 5.42×10(-4)) among ever-smokers.

Conclusions: Polymorphisms spanning previously associated lung function genes did not show strong evidence for association with lung function measures in the SpiroMeta consortium population. Common SERPINA1 polymorphisms may affect FEV(1) among smokers in the general population.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0019382PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3098839PMC
November 2011

The influence of exercise on morphological and neurochemical properties of neurons in rat nodose ganglia.

Neurosci Lett 2011 Feb 15;490(1):36-40. Epub 2010 Dec 15.

Department of Anatomy, Histology and Embryology, University of Split, School of Medicine, Šoltanska 2, 21000 Split, Croatia.

Physical exercise can induce immunohistochemical changes and cell proliferation in the hippocampus. One of the main effects of prolonged exercise is resting bradycardia, most probably caused by enhanced vagal activity. To investigate whether physical exercise can cause neurochemical and morphological changes in vagal afferent neurons, we performed immunohistochemical studies of nodose neurons using isolectin B4 (IB4), 200-kDa neurofilament protein (N52) and calretinin in adult female rats. To distinguish subpopulations of neurons projecting to the left ventricle, we applied a Fast Blue patch to the epicardial surface of the left ventricle. Treadmill running for 8 weeks significantly increased the size of N52-positive cardiac projecting neurons. Furthermore, the proportion of IB4-positive neurons among all nodose ganglia neurons was significantly higher in trained animals. These data indicate that exercise leads to plastic changes in nodose ganglia neurons that may initiate changes of vagal activity caused by prolonged exercise.
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http://dx.doi.org/10.1016/j.neulet.2010.12.021DOI Listing
February 2011

New paradigm in training of undergraduate clinical skills: the NEPTUNE-CS project at the Split University School of Medicine.

Croat Med J 2010 Oct;51(5):373-80

Unit for Science and Research, University Hospital Split, 21 000 Split, Croatia.

Clinical skills' training is arguably the weakest point in medical schools' curriculum. This study briefly describes how we at the Split University School of Medicine cope with this problem. We consider that, over the last decades, a considerable advancement in teaching methodologies, tools, and assessment of students has been made. However, there are many unresolved issues, most notably: (i) the institutional value system, impeding the motivation of the teaching staff; (ii) lack of a strong mentoring system; (iii) organization, timing, and placement of training in the curriculum; (iv) lack of publications pertinent to training; and (v) unwillingness of patients to participate in student training. To improve the existing training models we suggest increased institutional awareness of obstacles, as well as willingness to develop mechanisms for increasing the motivation of faculty. It is necessary to introduce changes in the structure and timing of training and to complement it with a catalog, practicum, and portfolio of clinical skills. At Split University School of Medicine, we developed a new paradigm aimed to improve the teaching of clinical skills called "Neptune-CSS," which stands for New Paradigm in Training of Undergraduate Clinical Skills in Split.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2969132PMC
http://dx.doi.org/10.3325/cmj.2010.51.373DOI Listing
October 2010

Histological differences in healing following experimental transmural infarction in rats.

Histol Histopathol 2010 12;25(12):1507-17

Department of Anatomy, Histology and Embryology, School of Medicine, Split, Croatia.

Mechanisms of cardiac regeneration following transmural myocardial infarction were analysed in rat hearts using immunohistochemistry for a-SMA, caspase-3, Ki-67 and nestin markers. Seven weeks after experimental myocardial infarction, two different types of healing processes were revealed in rats with and without aneurysmatic bulging of the left ventricular wall. Besides thinning of the ventricular wall, three zones characterized both types of scars: the scar zone (divided into central and peripheral parts), the peri-infarct zone and the border zone. The main difference between the types of scars was the presence of a central necrotic zone inside the aneurysmatic wall, while connective tissue with myofibroblasts characterized the same zone in non-bulging wall. Apoptotic caspase-3 positive cells were found in the granulation tissue of the border zone in aneurysmatic scar, while in non-bulging scar they characterized all three zones. Proliferating Ki-67 positive cells displayed reverse expression pattern compared to apoptotic cells. Quantification of a-SMA positive cells revealed 60% a-SMA positive cells inside the central part of the aneurysmatic scar zone and 39% in invaginating areas, versus 19% in non-invaginating areas of the peripheral zone, but only 30% in the peripheral part of the non-bulging scar zone. Nestin positive cells were found in both types of scars, but with different distribution. These results suggest that even seven weeks after myocardial infarction, the healing processes in non-bulging scars are in chronic phase, while aneurysmatic scars are still in subacute phase. Histological differences in scar healing might be important for functional properties of the heart wall and for heart recovery prognosis.
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http://dx.doi.org/10.14670/HH-25.1507DOI Listing
December 2010

Immunohistochemical characteristics of neurons in nodose ganglia projecting to the different chambers of the rat heart.

Auton Neurosci 2010 Jun 27;155(1-2):33-8. Epub 2010 Jan 27.

Department of Anatomy, Histology and Embryology, School of Medicine in Split, Soltanska 2, 21000 Split, Croatia.

Despite the contribution of nodose ganglia neurons to the innervation of the heart being the subject of several studies, specific neuronal subpopulations innervating the four different chambers of the heart have not been distinguished. In our study, the application of Fast Blue-loaded patch to the epicardial surface of different chambers of the rat heart (the right or left atrium or the right or left ventricle) resulted in labeling of discrete populations of immunohistochemically diverse neurons. About one half (55%) of these neurons showed immunoreactivity for the 200-kDa neurofilament protein (marker of myelinated neurons), with a higher proportion of positive staining among neurons projecting to the left than to the right ventricle. Isolectin B4 immunoreactivity (characteristic for a subset of nonmyelinated non-peptidergic neurons) was more abundant among neurons projecting to the right side of the heart (right atria and right ventricles) compared to the left side (23% vs. 16%). Calretinin immunoreactivity (possible marker of mechanosensitive neurons) was significantly higher among neurons projecting to the ventricles than among those projecting to atria (36% vs. 11%). These findings reveal that chambers of the rat heart are innervated with immunohistochemically different subpopulations of neurons from the nodose ganglia.
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http://dx.doi.org/10.1016/j.autneu.2010.01.001DOI Listing
June 2010

Characterization of spinal afferent neurons projecting to different chambers of the rat heart.

Neurosci Lett 2010 Jan 16;469(3):314-8. Epub 2009 Dec 16.

School of Medicine in Split, Department of Anatomy, Histology and Embriology, Soltanska 2, 21000 Split, Croatia.

The pattern of distribution of spinal afferent neurons (among dorsal root ganglia-DRGs) that project to anatomically and functionally different chambers of the rat heart, as well as their morphological and neurochemical characteristics were investigated. Retrograde tracing using a patch loaded with Fast blue (FB) was applied to all four chambers of the rat heart and labeled cardiac spinal afferents were characterized by using three neurochemical markers. The majority of cardiac projecting neurons were found from T1 to T4 DRGs, whereas the peak was at T2 DRG. There was no difference in the total number of FB-labeled neurons located in ipsilateral and contralateral DRGs regardless of the chambers marked with the patch. However, significantly more FB-labeled neurons projected to the ventricles compared to the atria (859 vs. 715). The proportion of isolectin B(4) binding in FB-labeled neurons was equal among all neurons projecting to different heart chambers (2.4%). Neurofilament 200 positivity was found in greater proportions in DRG neurons projecting to the left side of the heart, whereas calretinin-immunoreactivity was mostly represented in neurons projecting to the left atrium. Spinal afferent neurons projecting to different chambers of the rat heart exhibit a variety of neurochemical phenotypes depending on binding capacity for isolectin B(4) and immunoreactivity for neurofilament 200 and calretinin, and thus represent important baseline data for future studies.
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http://dx.doi.org/10.1016/j.neulet.2009.12.016DOI Listing
January 2010

Genome-wide association study identifies five loci associated with lung function.

Nat Genet 2010 Jan 13;42(1):36-44. Epub 2009 Dec 13.

Departments of Health Sciences and Genetics, Adrian Building, University of Leicester, Leicester, UK.

Pulmonary function measures are heritable traits that predict morbidity and mortality and define chronic obstructive pulmonary disease (COPD). We tested genome-wide association with forced expiratory volume in 1 s (FEV(1)) and the ratio of FEV(1) to forced vital capacity (FVC) in the SpiroMeta consortium (n = 20,288 individuals of European ancestry). We conducted a meta-analysis of top signals with data from direct genotyping (n < or = 32,184 additional individuals) and in silico summary association data from the CHARGE Consortium (n = 21,209) and the Health 2000 survey (n < or = 883). We confirmed the reported locus at 4q31 and identified associations with FEV(1) or FEV(1)/FVC and common variants at five additional loci: 2q35 in TNS1 (P = 1.11 x 10(-12)), 4q24 in GSTCD (2.18 x 10(-23)), 5q33 in HTR4 (P = 4.29 x 10(-9)), 6p21 in AGER (P = 3.07 x 10(-15)) and 15q23 in THSD4 (P = 7.24 x 10(-15)). mRNA analyses showed expression of TNS1, GSTCD, AGER, HTR4 and THSD4 in human lung tissue. These associations offer mechanistic insight into pulmonary function regulation and indicate potential targets for interventions to alleviate respiratory disease.
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http://dx.doi.org/10.1038/ng.501DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2862965PMC
January 2010

Daily mini quizzes as means for improving student performance in anatomy course.

Croat Med J 2009 Feb;50(1):55-60

Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Soltanska 2, 21000 Split, Croatia.

Aim: To evaluate daily-written 10-question quizzes in a medical anatomy course as a way to integrate assessment into the course and to evaluate their effect on the course success.

Methods: Students answering correctly 8/10 or more questions were awarded 0.5 points per quiz. There were 34 quizzes with a maximum point score 17. Measurable outcomes of academic progress in anatomy course (pass rates on 4 examination terms, total pass rate, and average marks) were calculated, and 2007/08 academic year was compared with the previous academic year in which daily written quizzes were not a part of the course. The relationship between cumulative points on daily quizzes and 3 components of the final examination (written, practical, and oral) for 2007/08 academic year was assessed by non-parametric correlation testing.

Results: Individual scores on quizzes ranged from 1.5 to 13.5 points. There was a positive correlation between scores on quizzes and grades on 3 components of the final examination: written (Spearman rho=0.784, P<0.001, n=79), practical (Spearman rho=0.342, P<0.002, n=79), and oral (Spearman rho=0.683, P<0.001, n=79) part. Compared with students in the previous academic year, students attending the course with daily quizzes significantly improved their academic achievement, expressed as the pass rate at the first examination term (39% vs 62%, respectively, chi(2) test, P=0.006, ) and the average course grade (2.71+/-1.08 vs 3.38+/-1.26, respectively; t test, P<0.001).

Conclusion: Despite their frequency and possible associated stress, daily quizzes were associated with better academic success in the anatomy course.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2657562PMC
http://dx.doi.org/10.3325/cmj.2009.50.55DOI Listing
February 2009