Antihyperlipidemic potential of dietary supplementation with carnosine in high-fat diet-fed rats.

OBJECTIVE: The aim of this study was to investigate the hypolipidemic effects of carnosine and a commercial carnosine supplement on lipid status, liver and kidney function, and inflammation associated with dyslipidemia in rats with high-fat diet-induced hyperlipidemia. MATERIALS AND METHODS: The study was conducted on adult male Wistar rats, divided into control and experimental groups. Animals were kept in standard laboratory conditions and according to groups were treated with saline, carnosine, carnosine dietary supplement, simvastatin, and their combinations. All substances were prepared fresh every day and used by oral gavage. RESULTS: Treatment with a carnosine-based supplement significantly improved total and LDL cholesterol levels in serum, especially in the combination with simvastatin as a conventional drug in dyslipidemia treatment. The effect of carnosine on the metabolism of triglycerides was not as evident as in the case of cholesterol. Nevertheless, the values of the atherogenic index showed that the combinations of carnosine and carnosine supplement with simvastatin were the most effective in lowering this comprehensive lipid index. Dietary carnosine supplementation resulted also in anti-inflammatory effects, as demonstrated by immunohistochemical analyses. Besides, the good safety profile of carnosine in terms of its effect on liver and kidney functions was also confirmed. CONCLUSIONS: The use of carnosine supplements in preventing and/or treatment of metabolic disorders requires further investigations into the mechanisms of action and potential interactions with conventional therapy.

Body mass index trend as a new parameter for evaluating children's nutritional status.

OBJECTIVES: The objective was to identify a body mass index (BMI) trend as a new indicator and predictor of children's nutritional status, replacing absolute BMI, and to demonstrate that a local BMI trend is more appropriate for this purpose than a national (or international) BMI trend. STUDY DESIGN: An entire school year group of primary school children were subject to a longitudinal 8-year cohort study. METHODS: BMI was measured three times during primary school education-on enrolment to primary school, then in the fifth and eighth grades. The BMI values obtained were used to create gender- and age-based percentile curves for children born in 1998/1999 (Osijek Percentile Curves (OPC) study) in the Osijek-Baranja County, Croatia. Every BMI result obtained was linked to one of the five percentile ranks (PRs) according to threshold percentiles 3, 10, 90 and 97; hence, the PRs were <3, ≥3 to <10, ≥10 to <90, ≥90 to <97 and ≥ 97. Percentiles and PRs were specified for each BMI value in a dual fashion, i.e. according to OPC and the entire Croatian territory (Croatian Percentile Curves (CROPC)-2008) cross-sectional study. After integrating the BMI values at all three time points, two BMI trends appeared-one based on the OPC criteria and one based on the CROPC-2008 criteria. Individual trends were grouped into three descriptive BMI categories: 'healthy', 'at risk' and 'unhealthy'. RESULTS: For the OPC and CROPC-2008, the 'healthy' trend characterised 2097 (78.72%) and 1975 (74.14%) children, respectively; the 'at risk' trend marked 434 (16.29%) and 458 (17.19%) children, respectively; and the 'unhealthy' trend was found in 133 (4.99%) and 231 (8.67%) children, respectively. The biggest difference between the OPC-based and CROPC-2008-based results was detected in the 'unhealthy' trend. According to CROPC-2008, there were almost twice as many children in that category. For the purpose of study design and comparing results obtained based on OPC and those obtained based on CROPC-2008, the BMI of one boy was used. His BMI values on starting primary school, in the 5th and in the 8th grades were 19.05 kg/m2, 26.11 kg/m2 and 27.11 kg/m2, respectively. Analysis of PRs based on OPC and CROPC-2008 reveals that the same boy was represented by different trends: '<90 <90 <97' and '<90 <97 <97', respectively. In terms of residence, girls from urban areas have higher BMI values than rural girls when starting primary school. CONCLUSION: This study suggests that a BMI trend could be a better indicator and predictor of children's nutritional status than absolute BMI. In addition, a cohort study is preferential to a cross-sectional study as a trend in an OPC-based study can be personalised. Also, local BMI reference values defined for each generation of children are more advantageous in this context than national BMI reference values.

Tool for decision-making regarding general evacuation during a rapid river flood.

OBJECTIVES: To propose a simple and effective tool for decision-making regarding general evacuation during a rapid river flood. STUDY DESIGN: Virtual testing of a tool in a real event. METHOD: A four-component tool was applied to build an alternative scenario of the catastrophic river flood in Obrenovac, Serbia, on May 2014. The components of this tool are: (1) the amount of precipitation above the 95th percentile of all previous measurements; (2) upstream river discharge above the 95th percentile of all previous measurements; (3) upstream river level above the 95th percentile of all previous measurements; and (4) worsening of the hydrometeorological situation in the following 48 h. RESULTS: In the early morning of 16 May 2014, a rapid river wave flooded 80% of the Obrenovac territory. There were 13 deaths due to drowning. Application of the study tool shows that these lives could have been saved, as the score to recommend general evacuation was reached 1 day before the flooding. The application of this tool to two previous great floods in Serbia shows that the score to recommend general evacuation was reached either 1 day before or on the onset of flash flooding. CONCLUSIONS: Due to its simplicity, this tool is universally applicable to facilitate decision-making regarding general evacuation during a rapid river flood, and it should be further tested in future similar catastrophes.

Docking-based preliminary study on the interactions of bile acids with drugs at the transporter level in intestinal bacteria.

OBJECTIVE: The aim of this study was to estimate the binding-affinities of different bile acids towards drug transporters in Lactobacillus acidophilus and Bifidobacterium longum in order to predict the influence of bile acids and probiotics interactions on drug pharmacokinetics. MATERIALS AND METHODS: In order to study interactions of bile acids with transporters of intestinal bacteria, molecular-docking step was performed, using SwissDock web-service. For the purpose of comparison, two natural bile acids, cholic acid (CA) and deoxycholic acid (DCA), and one semi-synthetic bile acid, 12-monoketocholic acid (MKC), were studied in parallel. The free-binding energy was used as the main criterion for ranking ligands. RESULTS: Studied bile acids exhibited different binding affinities towards bacterial transporters with MKC showing the most prominent effect. For the majority of studied transporters, the estimated affinities of bile acids decreased in the following order: MKC-CA-DCA. Namely, 38.7% of examined transport proteins gave the lowest free-binding energy with MKC. The weak inverse relationship between number of hydrogen bonds and estimated free-binding energies was revealed. CONCLUSIONS: The predominant effect of MKC for the majority of studied transport proteins suggests that keto group at carbon 12 of the steroid core has a significant influence on the properties of MKC and consequently, on interactions with membrane transporters. Present findings might have a role in the prediction of potential influence of bile acids and probiotics on drug pharmacokinetics.

Analgesic effects of rosemary essential oil and its interactions with codeine and paracetamol in mice.

OBJECTIVE: The use of herbal medicinal products in the management of pain has been increasing steadily in recent years, often in combination with conventional analgesics, which can induce significant interactions. In traditional medicine, rosemary was used as mild analgesic, for relieving renal colic pain and dysmenorrhea. The aim of our study was to examine analgesic effects of rosemary essential oil and its pharmacodynamic interactions with codeine and paracetamol in mice. MATERIALS AND METHODS: The identification and quantification of chemical constituents of the essential oil isolated from air-dried aerial parts of rosemary were carried out by GC/FID and GC/MS. The hot plate test was performed on NMRI mice by placing them individually on hot plate and assessing their response to the thermal stimulus. RESULTS: In this research, we identified 29 chemical compounds of the studied rosemary essential oil, and the main constituents were 1,8-cineole, camphor, and α-pinene. Administration of investigated essential oil increased significantly the latency time of animal response to heat-induced pain between 20th and 50th minute of the test, when compared to saline-treated group. Rosemary essential oil in the dose of 20 mg/kg was shown to be more efficient than in the dose of 10 mg/kg, in combinations with both codeine and paracetamol. CONCLUSIONS: Our findings support the use of rosemary in the management of pain and indicate a therapeutic potential of rosemary essential oil in combination with analgesic drugs. The mechanisms involved in analgesic effects of rosemary essential oil and the potential influence on cytochromes and drug metabolism should be more in-depth investigated.

[Diurnal fluctuations of protein contents and the pH dependence of beta2-microglobulin stability in urine].

Diurnal fluctuations of protein excretion into urine and the effect of urinary pH on the urinary protein concentrations were studied in patients with various kidney diseases. The diurnal kinetics of gamma-immunoglobulin, transferrin, albumin, alpha1-microglobulin, gamma-immunoglobulin light chains, and the retinol-binding protein proved to positively correlate with the diurnal fluctuations of proteinuria and to negatively correlate with urinary pH. Diurnal changes in urinary beta2-microglobulin content did not correlate with those of any other protein. Oral bicarbonate intake alkalinized the urine, increased the urinary beta2-microglobulin content, and led to a direct correlation between beta2-microglobulin excretion and excretion of other low-molecular proteins. Thus, proteinuria, single protein excretion, and urinary pH displayed diurnal rhythmicity in the patients; beta2-microglobulin was unstable in acid urine and its urinary level depended on the urinary pH.

Green fluorescent protein-tagged sarco(endo)plasmic reticulum Ca2+-ATPase overexpression in Paramecium cells: isoforms, subcellular localization, biogenesis of cortical calcium stores and functional aspects.

We have followed the time-dependent transfection of Paramecium cells with a vector containing the gene of green fluorescent protein (GFP) attached to the C-terminus of the PtSERCA1 gene. The outlines of alveolar sacs (ASs) are labelled, as is the endoplasmic reticulum (ER) throughout the cell. When GFP fluorescence is compared with previous anti-PtSERCA1 antibody labelling, the much wider distribution of GFP (ER+ASs) indicates that only a small amount of SERCA molecules is normally retained in the ER. A second isoform, PtSERCA2, also occurs and its C-terminal GFP-tagging results in the same distribution pattern. However, when GFP is inserted in the major cytoplasmic loop, PtSERCA1 and two fusion proteins are mostly retained in the ER, probably because of the presence of the overt C-terminal KKXX ER-retention signal and/or masking of a signal for transfer into ASs. On the overall cell surface, new SERCA molecules seem to be permanently delivered from the ER to ASs by vesicle transport, whereas in the fission zone of dividing cells ASs may form anew. In cells overexpressing PtSERCA1 (with C-terminal GFP) in ASs, [Ca2+]i regulation during exocytosis is not significantly different from controls, probably because their Ca2+ pump has to mediate only slow reuptake.

Microdomain arrangement of the SERCA-type Ca2+ pump (Ca2+-ATPase) in subplasmalemmal calcium stores of paramecium cells.

We localized SERCA pumps to the inner region of alveolar sac membranes, facing the cell interior, by combining ultrastructural and biochemical methods. Immunogold labeling largely predominated in the inner alveolar sac region which displayed aggregates of intramembrane particles (IMPs). On image analysis, these represented oligomeric arrangements of approximately 8-nm large IMP subunits, suggesting formation of SERCA aggregates (as known from sarcoplasmic reticulum). We found not only monomers of typical molecular size ( approximately 106 kD) but also oligomeric forms on Western blots (using anti-SERCA antibodies, also against endogenous SERCA from alveolar sacs) and on electrophoresis gelautoradiographs of 32P-labeled phosphoenzyme intermediates. Selective enrichment of SERCA-pump molecules in the inner alveolar sac membrane region may eliminate Ca2+ after centripetal spread observed during exocytosis activation, while the plasmalemmal Ca2+ pump may maintain or reestablish [Ca2+] in the narrow subplasmalemmal space between the outer alveolar sac membrane region and the cell membrane. We show for the first time the microzonal arrangement of SERCA molecules in a Ca2+ store of a secretory system, an intensely discussed issue in stimulus-secretion coupling research.

Molecular characterization of a sarco(endo)plasmic reticulum Ca2+-ATPase gene from Paramecium tetraurelia and localization of its gene product to sub-plasmalemmal calcium stores.

A cDNA encoding the gene for a sarco(endo)plasmic reticulum-type Ca2+-ATPase (SERCA) was isolated from a cDNA library of Paramecium tetraurelia by using degenerated primers according to conserved domains of SERCA-type ATPases. The identified nucleotide sequence (PtSERCA) is 3114 nucleotides in length with an open reading frame of 1037 amino acids. An intron of only 22 nucleotides occurs. Homology searches for the deduced amino acid sequence revealed 38-49% similarity to SERCA-type ATPases from organisms ranging from protozoans to mammals, with no more similarity to some parasitic protozoa of the same phylum. The calculated molecular mass of the encoded protein is 114.7 kDa. It contains the typical 10 transmembrane domains of SERCA-type ATPases and other conserved domains, such as the phosphorylation site and the ATP binding site. However, there are no binding sites for phospholamban and thapsigargin present in the PtSERCA. Antibodies raised against a cytoplasmic loop peptide between the phosphorylation site and the ATP binding site recognize on Western blots a protein of 106 kDa, exclusively in the fraction of sub-plasmalemmal calcium stores ('alveolar sacs'). In immunofluorescence studies the antibodies show labelling exclusively in the cell cortex of permeabilized cells in a pattern characteristic of the arrangement of alveolar sacs. When alveolar sacs where tested for phosphoenzyme-intermediate formation a phosphoprotein of the same molecular mass (106 kDa) could be identified.

[Peritoneal dialysis in patients with polycystic kidneys]. / Peritoneumska dijaliza kod bolesnika s policisticnim bubregom.

Polycystic kidney disease is a relative contraindication for active treatment with peritoneal dialysis (PD). We have treated 10 end stage kidney disease patients with PD, 4 with intermittent PD and 6 with CAPD. PD was successful in most of these patients. The following complications recurred: a marked increase in arterial blood pressure occurred in one patient, epileptiform fits appeared three times in a patient due to hypocalcemia; renal cancer was documented in a patient after two months on PD. Our results support the view that in selected cases peritoneal dialysis could be successfully used in end-stage polycystic kidney disease.

[Epidemiology of Lyme borreliosis]. / Epidemiologija Lyme borreliosis.

Our prospective study covered 1093 patients with epidemiological features and demonstrable clinical manifestations of Lyme borreliosis from 1987 to 1992. Basic criteria for the inclusion of our patients in the study were: tick bite, erythema migrans and antibodies to B. burgdorferi in each patient's serum. Various clinical manifestations were found un 1034 (94.6%) patients out of whom 829 (80.2%) had a history of tick bite, 901 (87.1%) had erythema migrans and 217 (21.0%) had serologic confirmation of the diagnosis. Inapparent infection was discovered in 59 (5.4%) patients. Other Types of skin changes (II and III stage) were found in 123 (11.9%) patients. The disease resulted in neurologic, joint and cardiac manifestations in somewhat lower percentage of patients (6.3%, 6.6% and 1.5%, respectively). In the three-year period (1990-1992) microscopic examination of the ticks (Ixodes ricinus) collected at several green areas of Belgrade confirmed the presence of B. burgdorferi in 29.1 per cent.

Partial lecithin:cholesterol acyltransferase (LCAT) deficiency in Balkan endemic nephropathy.

The role of lipid abnormalities has been also implicated in the progression of renal diseases. The search for lipid abnormalities in Balkan endemic nephropathy (BEN) has roused sporadic interest and has not been fully elucidated. This study was performed in 54 healthy subjects from the families affected with BEN (group A), 18 members from non-affected families living in the same location (group B), and 25 control subjects (group C). Lipid profiles and lecithin:cholesterol acyltransferase (LCAT) were determined in each subject. The most striking distinction between the groups was that of the LCAT activity, which was abnormally low in group A (39 +/- 2), significantly different (P less than 0.0001) from that of the other groups. Thirty individuals from group A were those accounting for the low LCAT activity (A1). This group had a significantly lower total cholesterol and free cholesterol than all of the other subjects. The entire group A subjects had a significantly lower percentage of free cholesterol than the other two groups. There was no significant difference in HDL cholesterol between any of the groups, but group A1 had significantly higher HDL than group C (P less than 0.04). What emerges from our study is that a certain proportion of subjects from BEN families have a peculiar form of lipid abnormalities associated with an abnormal LCAT activity. At present we have no explanation for these findings. We believe that these changes may have an important role in the pathogenesis of BEN.

Beta 2-microglobulin in patients with Balkan nephropathy and in healthy members of their families.

The aim of this study was to investigate serum levels, urinary excretion and in vitro peripheral blood mononuclear cell (PBMC) production of beta 2-microglobulin in patients with Balkan nephropathy and their families. Increased urinary beta 2-microglobulin excretion was found in Balkan nephropathy, chronic pyelonephritis and glomerulonephritis patients, being highest in the first group. The serum level of beta 2-microglobulin in Balkan nephropathy patients correlated with residual kidney function. Synthesis of beta 2-microglobulin by PBMC, untreated or stimulated by PHA, was not increased in Balkan nephropathy patients or their healthy family members compared to the control group of healthy persons living outside of an endemic region. This study has shown that the increased serum beta 2-microglobulin level in Balkan nephropathy patients is the consequence of the glomerular filtration rate (GFR) reduction. Urinary beta 2-microglobulin excretion was found increased not only in patients but in some healthy members of nephropathic families. beta 2-microglobulin therefore can serve as a marker of the early tubular damage in Balkan nephropathy. However, urinary beta 2-microglobulin is not specific for Balkan nephropathy, lacking specificity required for screening purposes. The different patterns of serum and urinary beta 2-microglobulin, and other urinary proteins, in patients with Balkan nephropathy from patients with chronic pyelonephritis and glomerulonephritis favor the opinion that Balkan nephropathy is a separate clinical entity.

[Possibilities in the practical use of computers in the selection of candidates for kidney transplantation]. / O nekim mogucnostima prakticne primene racunara u izboru kandidata za transplantaciju bubrega.

This paper points out some problems appearing in the selection of candidates for the cadaveric kidney transplantation treats basic issues regarding medical and other references important for making decision in the process of candidate selection, emphasizes the significance of data base for prepared candidates and presents the specific example of the applications of APLLE II C in the practice of the clinic for nephrology. The data for each candidate are grouped in two sections: A. PERSONAL DATA 1. Family name and surname 2. Year of birth 3. Place of residence 4. Address 5. Telephone number 6. Register number of case history 7. Date of enlisting B. BASIC MEDICAL DATA 1. Blood type 2. HLA tipization of tissue: A, B, C, DR 3. Cytotoxic antibodies 4. Urgency priority 5. Date of eventual previous transpl. 6. Special remarks This mode of Candidate selection provides the availability of all relevant data at one place: objectivity and high efficiency. Creation of such data base in every Yugoslav Kidney Transplantation Center would "at the time" mean the formation of YU Information System, thus providing a possibility of choice of the most appropriate cadaveric Kidney recipient on Yugoslav level at any time.