Prevalence of hospitalized patients with sarcoidosis in Croatia

The aim of this study was to investigate the prevalence of hospitalized patients of sarcoidosis in the Republic of Croatia, its distribution in relation to sex and age as well as its geographical distribution. The data on sarcoidosis patients hospitalized in Croatia in the last six years, from 1997 to 2002, were analyzed retrospectively. The prevalence of sarcoidosis patients hospitalised in the Republic of Croatia is 4.1/100,000. The prevalence among women is 4.7 and among men 3.5 per 100,000 persons, with a ratio of 1.4:1. The disease more frequently occurs in the regions with a continental climate than in the Mediterranean zone. The ratio of sarcoidosis patients in the continental zone to the Mediterranean zone is 1.5:1. It occurs predominantly among the adults. Over the investigated period, in our country we have not registered any case of sarcoidosis among children. It occurs more frequently at a younger age and therefore 44.5% of the patients with sarcoidosis were between 20 and 39 years of age, 40.1% were between 40 and 59 years of age and 15.3% were more than 60 years old.     

Modulatory effects of acid-sensing ion channels on action potential generation in hippocampal neurons

Extracellular acidification has been shown to generate action potentials (APs) in several types of neurons. In this study, we investigated the role of acid-sensing ion channels (ASICs) in acid-induced AP generation in brain neurons. ASICs are neuronal Na⁺ channels that belong to the epithelial Na⁺ channel/degenerin family and are transiently activated by a rapid drop in extracellular pH. We compared the pharmacological and biophysical properties of acid-induced AP generation with those of ASIC currents in cultured hippocampal neurons. Our results show that acid-induced AP generation in these neurons is essentially due to ASIC activation. We demonstrate for the first time that the probability of inducing APs correlates with current entry through ASICs. We also show that ASIC activation in combination with other excitatory stimuli can either facilitate AP generation or inhibit AP bursts, depending on the conditions. ASIC-mediated generation and modulation of APs can be induced by extracellular pH changes from 7.4 to slightly <7. Such local extracellular pH values may be reached by pH fluctuations due to normal neuronal activity. Furthermore, in the plasma membrane, ASICs are localized in close proximity to voltage-gated Na⁺ and K⁺ channels, providing the conditions necessary for the transduction of local pH changes into electrical signals. cellular excitability; neuronal signaling; pH     

Structural characterization of extracellular lipase from Streptomyces rimosus: assignment of disulfide bridge pattern by mass spectrometry

The cloning, sequencing and high-level expression of the gene encoding extracellular lipase from Streptomyces rimosus R6-554W have been recently described, and the primary structure of this gene product was deduced using a bioinformatic approach. In this study, capillary electrophoresis-on-the-chip and mass spectrometry were used to characterize native and overexpressed extracellular lipase protein from S. rimosus . The exact molecular mass of the wild-type and the overexpressed lipase, determined by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry, were in excellent agreement (Deltam=0.11 Da and Deltam=0.26 Da, respectively) with a value of 24165.76 Da calculated from the structure deduced from the nucleotide sequence, considering the mature enzyme with all six cysteines forming disulfide bridges. The primary structure derived from the nucleotide sequence was completely verified using a combination of tryptic digestion and formic acid cleavage of the protein, followed by peptide mass fingerprinting. Selected peptides were further investigated by MALDI low-energy collision-induced dissociation hybrid tandem mass spectrometry, allowing the unambiguous determination of their predicted amino acid sequence. No post-translational modifications of mature S. rimosus lipase were detected. Comparison of the peptide mass fingerprints from the reduced and non-reduced overexpressed enzyme unequivocally revealed three intramolecular disulfide bonds with the following linkages: C27-C52, C93-C101 and C151-C198.     

Potentiation of dietary restriction-induced lifespan extension by polyphenols

Dietary restriction (DR) extends lifespan across multiple species including mouse. Antioxidant plant extracts rich in polyphenols have also been shown to increase lifespan. We hypothesized that polyphenols might potentiate DR-induced lifespan extension. Twenty week old C57BL/6 mice were placed on one of three diets: continuous feeding (control), alternate day chow (Intermittent fed, IF), or IF supplemented with polyphenol antioxidants (PAO) from blueberry, pomegranate, and green tea extracts (IF+PAO). Both IF and IF+PAO groups outlived the control group and the IF+PAO group outlived the IF group (all p<0.001). In the brain, IF induced the expression of inflammatory genes and p38 MAPK phosphorylation, while the addition of PAO reduced brain inflammatory gene expression and p38 MAPK phosphorylation. Our data indicate that while IF overall promotes longevity, some aspects of IF-induced stress may paradoxically lessen this effect. Polyphenol compounds, in turn, may potentiate IF-induced longevity by minimizing specific components of IF-induced cell stress.     

Gene expression and activity analysis of the first thermophilic U32 peptidase

Peptidase family U32 is one of the few whose catalytic type and structure has not yet been described. It is generally accepted that U32 peptidases represent putative collagenases and contribute to the pathogenicity of some bacteria. Meanwhile, U32 peptidases are also found in nonpathogenic bacteria including thermophiles and hyperthermophiles. Here we report cloning of the U32.002 peptidase gene from thermophilic Geobacillus thermoleovorans DSM 15325 and demonstrate expression and characterization of the recombinant protein. It has been determined that U32.002 peptidase is constitutively expressed in the cells of thermophilic G. thermoleovorans DSM 15325. The recombinant oligomeric enzyme showed its activity only against heat-treated collagen. It was unable to degrade albumin, casein, elastin, gelatine and keratin. In contrast to this, the monomeric recombinant protein showed no activity at all. This paper is the first report about the thermophilic U32 peptidase. As the thermophilic bacteria are non-pathogenic, the role of constitutively expressed extracellular collagenolytic U32 peptidase in these bacteria is unclear.     

Genome size and chromosome number in Echinops (Asteraceae, Cardueae) in the Aegean and Balkan regions: technical aspects of nuclear DNA amount assessment and genome evolution in a phylogenetic frame

This work focuses on the representatives of genus Echinops (Asteraceae, Cardueae) in the Aegean and Balkan regions, from the perspective of their genome evolution. Chromosome numbers were determined by orcein staining in 14 populations of nine taxa, and DNA contents were assessed by flow cytometry in 24 populations of nine taxa. A molecular phylogeny based on the internal transcribed spacer (ITS) and trnL-trnF and including first sequences for two taxa (Echinops sphaerocephalus subsp. taygeteus and E.?spinosissimus subsp. neumayeri) provided a framework for discussing genome changes. From a methodological point of view, similar C-DNA value estimates were obtained when measuring, for a same population, fresh leaves from adult plants collected in the field and from cultivated seedlings. Conversely, despite giving the appearance of being correct (e.g., low coefficient of variation), genome size assessed using silica gel-preserved material differs significantly from values obtained for the same populations with fresh material. Nevertheless, silica gel-preserved material may still provide rough estimates of genome size for, e.g., inferring ploidy level. Suitable—non-silica gel-based—DNA amounts assessed for 23 populations range from 2C?=?6.52?pg (E.?spinosissimus subsp. neumayeri) to 2C?=?9.37?pg (E.?bannaticus). Chromosome counts were established for the first time for Echinops graecus (2n?=?32), E.?sphaerocephalus subsp. albidus (2n?=?32), E.?sphaerocephalus subsp. taygeteus (2n?=?ca.?30), and E.?spinosissimus subsp. neumayeri (2n?=?28). Genome size and chromosome number are confirmed as crucial parameters for deciphering lineage diversification within the genus Echinops.     

Optical aspect of deformation analysis in the bone-denture complex

The aim of this study was to register and measure any deformation of mandible models under load. The method for full field measurement of strain is done by using the ARAMIS three-dimensional image correlation system. The system uses two digital cameras that provide a synchronized stereo view of the specimen and the results show the complete strain field during the tests. The biggest deformation values were just under the working force of the biggest intensity 500 N, and for the region of the lower second premolar the deformation is 625 microm. The following study is presented that highlight the use of stereometric measuring system for modern research. It is shown that this measuring methodology can capture the trends of the experiments.     

Relationship between the training of young recruits and values of bite forces

Analysis of masticatory function is the basis of clinical work in almost all fields of dentistry. Bite forces are the expression and measure of masticatory function. Physical training has an effect on the development of functional ability, motoric ability of the organism and the formation of desired physical proportions. The purpose of this study was to examine the association between physical fitness and bite force values. Because of strictly defined regulations in the army with regard to training and nutrition, Croatian Army recruits were ideal examinees for this examination. The examinees were 135 recruits. Bite forces were measured on three places (area of the central incisors, left and right in the area of the first molars) before and after three-months of training. Of all the examinees, 108 had increased their body weight, 12 had decreased it and 15 had not changed their body weight. The median of measured forces in the recruits prior to training was 291 N in the right (lateral quadrant), 285.5 N in the left lateral quadrant and 205 N in the anterior area. After training the median of measured forces in the right quadrant was 312 N, in the left 313 N and in the anterior area 216 N Greater bite forces after training on all measured places were statistically proved. Increased activity of masticatory muscles can have the same effect on the values of bite forces as bite training. There are few data on the correlation between physical muscles and values of bite forces. The results of those studies are doubtful. In this study, after three months of conditional training, the body mass of the recruits had increased and they expressed greater values of bite forces. However, correlation between body mass and bite forces cannot be proved with certainty.     

Unexpected sudden death due to recreational swimming and diving in men in Croatia in a 14-year period

The article deals with 17 sudden deaths which occurred during recreational swimming and diving in men in Croatia in a 14-year period: from January 1, 1998 to December 31, 2011. The sample is taken out from the total number of 61 sudden deaths in men during or immediately after sport or recreational exercise. Included are also sudden deaths of 8 foreigners spending holidays at the Croatian Adriatic Coast. In all of them forensic medicine autopsy was done. Thirteen males from Croatia died during recreational swimming. Three of them were aged 15-29 yrs: one had signs of hypertrophic cardiomyopathy, the second suffered from chronic myopericarditis with left ventricular aneurysm, and the third had cardiomegaly and blood alcohol level of 1.7 per thousand. Five were aged 30-64 yrs: four of them have suffered from coronary atherosclerosis and left ventricular hypertrophy of 15-18-18-22 mm, and one with left ventricular hypertrophy drowned suddenly, probably because of malignant ventricular arrhythmia. The fifth suffered stroke and drowned. Five elderly men, aged 65-85 yrs, have suffered from coronary atherosclerosis, myocardial fibrosis or myocardial scars, and three of them had left ventricular hypertrophy of 19 mm. Four males died during recreational diving. One aged 26yrs drowned, at autopsy he had left ventricular hypertrophy of 17 mm. Three males were middle-aged: two had coronary atherosclerosis, two of them had a severe degree of coronary atherosclerosis and one had coronary atherosclerosis of medium degree but with myocardial fibrosis and left ventricular hypertrophy of 18 mm. Seven male foreigners died, five of them during swimming: two aged 30-64 and two aged 65-85. They all have had coronary atherosclerosis: one of them had an acute myocardial infarction of the posterior wall, and one hypertrophic cardiomyopathy as well. One middle-aged and one elderly man died during diving, and both had an acute myocardial infarction of the posterior wall. One elderly foreign woman died during swimming, she had coronary atherosclerosis and a myocardial scar. In Croatia, death rate during both swimming and diving in men aged 15-29 years amounted to 0.63/1,000.000 (p=1.0000); in those aged 30-64 it reached 0.56/1,000.000 (p=0.3698), and in those aged 65-85 it was 1.41/1,000.000 (p=0.1849). The death rate during swimming in men aged 15-29 amounted to 1.47/1,000.000 (p=0.9864), in men aged 30-64 it reached 0.35/1,000.000 (p=0.2245), and in those aged 65-85 it was 1.41/1,000.000 (the difference is significant, p=0.0472). The death rate during diving in men aged 15-29 was 0.16/1,000.000, and in men aged 30-64 the observed rate was 0.21/1,000.000 (p=1.0000).