MicroRNA-target pairs in human renal epithelial cells treated with transforming growth factor β1: a novel role of miR-382

We reported previously an approach for identifying microRNA (miRNA)-target pairs by combining miRNA and proteomic analyses. The approach was applied in the present study to examine human renal epithelial cells treated with transforming growth factor β1 (TGFβ1), a model of epithelial–mesenchymal transition important for the development of renal interstitial fibrosis. Treatment of human renal epithelial cells with TGFβ1 resulted in upregulation of 16 miRNAs and 18 proteins and downregulation of 17 miRNAs and 16 proteins. Of the miRNAs and proteins that exhibited reciprocal changes in expression, 77 pairs met the sequence criteria for miRNA–target interactions. Knockdown of miR-382, which was up-regulated by TGFβ1, attenuated TGFβ1-induced loss of the epithelial marker E-cadherin. miR-382 was confirmed by 3′-untranslated region reporter assay to target five genes that were downregulated at the protein level by TGFβ1, including superoxide dismutase 2 (SOD2). Knockdown of miR-382 attenuated TGFβ1-induced downregulation of SOD2. Overexpression of SOD2 ameliorated TGFβ1-induced loss of the epithelial marker. The study provided experimental evidence in the form of reciprocal expression at the protein level for a large number of predicted miRNA-target pairs and discovered a novel role of miR-382 and SOD2 in the loss of epithelial characteristics induced by TGFβ1.     

Rocky-shore unconformities marking the base of Badenian (Middle Miocene) transgressions on Mt. Medvednica basement (North Croatian Basin,Central Paratethys)

Badenian (Middle Miocene) transgressive deepening-upward successions located in the NE part of Mt. Medvednica (North Croatian Basin, Pannonian Basin System) unconformably overlie Mesozoic basement. Triassic and Upper Cretaceous limestone pebbles, cobbles, and boulders of the Badenian basal conglomerates display abundant in situ bivalve borings of Gastrochaenolites and sponge borings of Entobia. This Gastrochaenolites-Entobia ichnoassemblage is related to the Entobia subichnofacies of the Trypanites ichnofacies, characterizing littoral rocky-shore environments (wave-cut platforms and marine transgressive surfaces with a low or null rate of sedimentation). Gastrochaenolites torpedo, Gastrochaenolites lapidicus, and Entobia recorded in Badenian basal conglomerates (compared with modern Northern Adriatic rocky-shore environments), enabled more precise palaeoenvironmental interpretations. The occurrence of G. torpedo (produced by lithophaginid bivalves) on all sides of individual limestone lithoclasts in the Gornje Ore?je basal conglomerate, coupled with truncation of the formation (possibly indicating multiphase colonization), reflect gravel transport, roll-over, overturning and erosion by wave action in high-energy rocky-shore settings. Gornje Psarjevo-2 basal conglomerate boulders were probably not subjected to significant movement and abrasion, as suggested by good preservation of both G. lapidicus (potentially produced by gastrochaenid bivalves), associated G. torpedo, as well as abundant shallow Entobia borings. The Badenian Gastrochaenolites-Entobia ichnoassemblage also could be the result of a composite development. However, direct cross-cutting relationships between G. torpedo and G. lapidicus and/or Entobia were rarely observed. In addition, Badenian boring tracemakers might have coexisted at the same water depth. Northeast Mt. Medvednica Badenian successions probably formed during different Central Paratethys transgressive pulses (NN5 and NN6 Zones). However, exact timing of Badenian transgressions, stratigraphic correlations and tectono-eustatic implications are unresolved, due to sparsely integrated biostratigraphic and high-precision geochronological studies of Early–Middle Miocene North Croatian Basin deposits and due to the absence of a uniform biostratigraphic zonation and regional chronostratigraphic division of Central Paratethys.     

Quality of life after cardiac surgery

The aim of this study was to determine a health-related quality of life two or more years after a cardiac surgery using the MOS-36 item short-form health survey questionnaire. 330 members of "Croatian society of patients who underwent cardiac surgery" were asked and 196 responders, 122 men and 74 women, were included in survey, all of them more than two years after surgery and older than fifty years of age. The answers were collected by phone. Women gained better results for Physical Functioning (p < 0.003) and men estimated better scores for psychical health (p < 0.001). Our patients gained some different results in respect to the Croatian sample of healthy people of the same age. They declared better General Health (p < 0.001), less Bodily Pain (p < 0.001), but inferior Role-Emotion (p < 0.001) and Social Functioning (p < 0.001). The estimations of QOL improving were comparable with the results of numerous shorter follow-ups.     

Pediatric urolithiasis in Croatia

A retrospective review was performed of the records of 148 Croatian children with urolithiasis treated between 1989 and 2003. The study evaluated age, gender, family history, clinical symptoms, location of stone, laboratory findings, stone composition, mode of treatment and compared our results with data from higher and lower socio-economic countries. The mean age of our patients was 9.38 years (10 months to 18 years). Thirty-seven children (25%) were less than 5 years (group 1), 44 (29.7%) were between 5 and 10 years (group 2) and 67 (45.3%) were older than 10 years of age (group 3). There were 60 girls and 88 boys with overall male to female ratio of 1.47. Abdominal pain (83%) and haematuria (59.5%) were the main symptoms in the groups 2 and 3. Urinary tract infection was predominant symptom in the group 1 (62.1%). Calculi were located in the kidney in 90 children (60.8%), in the ureter in 39 (26.4%), in the bladder in 8 (5.4%). Urinary tract anomalies with or without infection were associate with a greater frequency of urolithiasis in the youngest age group and hypercalciuria was predominant cause in children over 5. Stone analysis was performed in 80 children. Predominant constituent of stones was calcium oxalate (48.7%), followed by struvite (25%), calcium phosphate (13.7%), cystine (10%) and uric acid (1.2%). Calcium oxalate stones were most common in all age groups. Struvite stones were most prevalent in the children younger than 5 years of age. Most patients (33.1%) underwent surgery for removal of their calculi. In 31.8% of children stones were passed spontaneously and the highest spontaneous passage rate was in the group 3 (37.3%). Stone composition, location and etiology in Croatian children are similar to those in developed Western countries.     

Cerebrospinal fluid markers in differential diagnosis of Alzheimer's disease and vascular dementia

Alzheimer's disease (AD) and vascular dementia (VaD) are the two most common causes of dementia in old people. They remain difficult to differentiate in practice because of lack of sensitivity and specificity of current clinical diagnostic criteria. Recent molecular and cellular advancements indicate that the use of cerebrospinal fluid markers may improve early detection and differential diagnosis of AD. Our objective in this study was to determine diagnostic accuracy of three cerebrospinal (CSF) markers: total tau protein (t-tau), tau protein phosphorylated on threonine 181 (p-tau181) and tau protein phosphorylated on serine 199 (p-tau199). Using commercially available ELISA kits concentrations of t-tau, p-tau181 and p-tau199 were analyzed in 12 patients with probable AD, 9 patients with VaD and 12 NC subjects. The median levels of all three markers were significantly higher in AD group versus VaD and NC groups. However, when the sensitivity levels were set to 85% or higher, only t-tau and p-tau199 satisfied consensus recommendations (specificity more than 75%) when differentiating AD from VaD. In conclusion, our preliminary data on a small group of selected subjects suggest that the CSF t-tau and p-tau199 levels are useful markers for differentiating AD from VaD.     

Stable isotopic (δ13C and δ18O) signatures of biogenic calcretes marking discontinuity surfaces: a case study from Upper Cretaceous carbonates of central Dalmatia and eastern Istria,Croatia

Biogenic calcretes associated with a regional Cretaceous to Paleogene subaerial unconformity and an intraformational composite (polygenic) surface in Upper Cretaceous intra-platform peritidal successions in central Dalmatia and eastern Istria, Croatia (Adriatic-Dinaridic Carbonate Platform), were analyzed for their δ¹³C and δ¹⁸O signatures in order to provide insight into the conditions of subaerial exposure and calcrete development. The distinctly negative δ¹³C signatures of biogenic calcretes marking the regional subaerial unconformity differ considerably from the δ¹³C values of the host marine limestones. This indicates carbon isotope exchange of primary marine CaCO₃ with CO₂ released by root and rhizomicrobial respiration and subsequent precipitation of pedogenic calcrete. The range of δ¹³C (from ?13.1 to ?8.2 ‰ Vienna PeeDee Belemnite standard, VPDB) and δ¹⁸O (from ?10.1 to ?6.1 ‰ VPDB) values of calcretes are similar to those reported from calcretes elsewhere, and the δ¹³C values of biogenic calcretes with typical Microcodium aggregates (?13.1 to ?12.3 ‰ VPDB) at the ?ibenik locality are very close to, or at the lower limit of, values for soil carbonates formed in isotopic equilibrium with soil CO₂. These values are expected for authigenic pedogenic carbonates formed under the influence of C₃ plant communities, without influence from heavier carbon from pre-existing carbonate and lack of input of atmospheric CO₂. Such low δ¹³C values support the interpretation of Microcodium aggregates as being precipitated under a direct biological control within the soil, although the relationship between formation mechanisms and stable isotope signatures of Microcodium needs further investigation. The δ¹³C values (?4.4 to ?3.6 ‰ VPDB) of rhizogenic calcretes formed inside firmground Thalassinoides burrows of the composite surface at the ?ibenik locality are more negative than the δ¹³C values of the host marine limestones, which confirms that the composite surface went through a phase of meteoric pedo(dia)genesis. However, the overall δ¹³C values of calcretes are less negative than expected, which might reflect contamination from associated primary marine carbonate. This study represents the first detailed stable isotope investigation of calcretes from carbonate successions of the External Dinarides, and the results may be applied to discontinuities present in other shallow-water carbonate rock successions.     

Behaviour after cardiosurgery

Aim of this study was to estimate behavioral changes after cardiac surgery (CS). A questionnaire was sent by mail to 74 women and 122 men during May 2004 and the answers were collected by phone. For statistical analysis, the chi2-test was used. A common unhealthy habits before CS were inappropriate diet (80% of all responders), addictions to alcohol (62%) and nicotine (54%). Significantly fewer patients after CS continue with fatty diet (chi2 = 39.069; p < 0.001), smoking (chi2 = 90.286; p < 0.001) and alcohol drinking habits (chi2 = 60.667; p < 0.001). A significant worsening of sexual life (chi2 = 91.533; p < 0.001) and significantly less negative influence of weather changes (chi2 = 20.821; p < 0.001) was found after CS. Men drink more (chi2 = 16.299; p < 0.001) and smoke more (chi2 = 19.635; p < 0.001), have better sexual life (chi2 = 17.317; p < 0.001), fewer sleeping disorders (chi2 = 17.334; p < 0.001) and better oral health (chi2 = 38.632; p < 0.001) than women. These results can be useful for setting up preventive measures for CVD.