A new model to determine lipophilicity of 1,2-ethanediamine-N,N'-di-2-(3-cyclohexyl)propanoic acid and 1,3-propanediamine-N,N'-di-2-(3-cyclohexyl)propanoic acid derivatives with antiproliferative activity by combining shake flask procedure and UHPLC-MS method.

Fourteen compounds representing ester derivatives of (S,S)-1,2-ethanediamine-N,N'-di-2-(3-cyclohexyl) propanoic and (S,S)-1,3-propanediamine-N,N'-di-2-(3-cyclohexyl)propanoic acids, expressing antiproliferative activity in vitro were examined. The objective of this study was to determinate their lipophilicity data, and also to ensure a mathematical model for prediction lipophilicity data of potential in vivo metabolites and new derivatives of (S,S)-1,2-ethanediamine-N,N'-di-2-(3-cyclohexyl)propanoic acid, based on chromatographic parameters. Experimentally, lipophilicity data were obtained by a traditional shake flask procedure and an ultra-high performance liquid chromatographic tandem mass spectrometry (UHPLC-MS) method. A correlation between the partition coefficient n-octanol/water (logD7,4) and chromatographic data (CHI, 0), and also, between logD7,4 and retention time was investigated. A very good correlation (r2=0.8969) was found between lipophilicity parameters 0 and logD7,4 obtained using UHPLC-MS and shake flask methods: logD7,4 = (0.11±0.01)×0 + (1.25±0.20)×Nc - (9.19±1.18); statistical parameter F=47.84; significance of F = 3.74×10-6, Nc=number of C atoms between two amino groups (Nc=2 for 1,2-ethanediamine derivatives and Nc=3 for 1,3-propanediamine derivatives). The model predictivity power was determined by cross validation leave one out (LOO) technique, and expressed by the term Q2, was 0.89. The developed model has good predictivity power for prediction lipophilicity data of potential in vivo metabolites of the investigated compounds, such as novel 1,2-ethanediamine and 1,3-propanediamine N,N'-di-2-(3-cyclohexyl)propanoic acid derivatives. Also, the lipophilicity data obtained in the present study correlated with the antiproliferative activity of the investigated substances shown previously in in vitro studies.

[CARDIORENAL SYNDROME: CLINICAL FEATURES, EARLY DIAGNOSIS AND TREATMENT AT FAMILY MEDICINE].

The interdependent damage to the heart and kidney organ systems is defined as cardiorenal syndrome, a complex pathophysiological disorder of the heart and kidney in which acute or chronic dysfunction of one organ can lead to acute or chronic damage to the other. Identification and early diagnosis of some subtypes of cardiorenal syndrome very often begin at family physician office, however, the use of simple and reliable diagnostic procedures such as MICE score using ECG and biomarkers has not been implemented yet. The clinical picture, diagnosis and treatment vary according to the 5 cardiorenal syndrome subtypes, as described herein. Rational diagnosis of heart failure at family medicine office should include biomarkers (BNP and NT-pro BNP) before performing ultrasound of the heart, while for kidneys creatinine and estimated glomerular filtration rate are still in use, but not cysteine C and NGAL. Diagnostic procedure for suspected heart failure at family medicine office should include kidney function estimate and vice versa. Access to treatment of cardiorenal syndrome differs depending on the specialty to which the patient is referred first, i.e. consultant examination, cardiologist or nephrologist. A multidisciplinary approach to treatment of cardiorenal syndrome is still lacking.

[PREVENTION AND EARLY DETECTION OF COLORECTAL CANCER].

Colorectal cancer is a global problem worldwide because of its very high prevalence and mortality. Therefore, prevention of colorectal cancer and its early diagnosis is of great importance. In Croatia, the National Program for Colorectal Cancer has been carried out since 2007; however, the rate of response was about 18 percent, depending on the region. Such a great public health and social and economic problem requires multidisciplinary approach in which family physicians have an important role. The well spread and developed network of primary health care and the availability of family physicians to each inhabitant have not been sufficiently exploited, especially for such preventive activities where family physicians could supervise program implementation.

[APPROACH TO A PATIENT WITH DYSPEPSIA IN FAMILY MEDICINE PRACTICE].

Dyspepsia is a common symptom among patients in family medicine practice. The prevalence in adult population is about 40%. Two-thirds of patients have functional dyspepsia. Clinical assessment, diagnostic procedures and treatment of patients depend on the age, symptoms and Helicobacter pylori infection. In patients with dyspepsia, it is necessary to assess the potential impact of other concurrent diseases and medications that the patient regularly uses. Prompt or early endoscopy is recommended in patients with newly detected dyspepsia older than 50 and presenting with alarming symptoms. In persons younger than 50, the recommended strategy is 'test and treat'. In some patients, treatment is carried out by acid suppression. In patients failing to achieve success in treatment, further endoscopic diagnosis is indicated. Ultrasound diagnostics in primary care can significantly contribute to diagnostic evaluation and early treatment in patients with hepatobiliary and pancreas diseases presenting with symptoms of dyspepsia. Treatment of concurrent mental disorders can improve the symptoms of dyspepsia. Treatment of patients who do not respond to the recommended treatment strategies is a challenge for family physicians. Regular visits and psychotherapeutic support in these patients can reduce the level of anxiety and encourage the patient for treatment of psychological morbidity, as well as his efforts in healthy behavior.

Lack of mitochondrial DNA structure in Balkan donkey is consistent with a quick spread of the species after domestication.

A total of 132 mtDNA sequences from 10 Balkan donkey populations were analysed to ascertain their regional genetic structure and to contribute to the knowledge of the spreading of the species after domestication. The Balkan donkey sequences were compared with those from 40 Burkina Faso donkeys as an African outgroup to account for possible local Balkan scenarios. The 172 sequences gave 62 different haplotypes (55 in Balkan donkey). Virtually all the analysed populations had haplotypes assigned to either Clade 1 or Clade 2 even though the relative proportion of Clade 1 or 2 haplotypes differed across populations. Geographical maps constructed using factors computed via principal component analysis showed that the Balkan donkey populations are not spatially structured. AMOVA confirmed a lack of genetic structure in Balkan donkey mtDNA. Balkan populations were poorly differentiated (ΦST  = 0.071). Differentiation between the Balkan donkey and the African outgroup also was low. The lack of correspondence between geographical areas and maternal genetic structure is consistent with the hypothesis suggesting a very quick spread of the species after domestication. The current research illustrates the difficulties to trace routes of expansion in donkey, as the species has no geographical structure.

Brain and art: illustrations of the cerebral convolutions. A review.

BACKGROUND: Aesthetics and functional significance of the cerebral cortical relief gave us the idea to find out how often the convolutions are presented in fine art, and in which techniques, conceptual meaning and pathophysiological aspect. MATERIALS AND METHODS: We examined 27,614 art works created by 2,856 authors and presented in art literature, and in Google images search. RESULTS: The cerebral gyri were shown in 0.85% of the art works created by 2.35% of the authors. The concept of the brain was first mentioned in ancient Egypt some 3,700 years ago. The first artistic drawing of the convolutions was made by Leonardo da Vinci, and the first colour picture by an unknown Italian author. Rembrandt van Rijn was the first to paint the gyri. Dozens of modern authors, who are professional artists, medical experts or designers, presented the cerebralc onvolutions in drawings, paintings, digital works or sculptures, with various aesthetic, symbolic and metaphorical connotation. Some artistic compositions and natural forms show a gyral pattern. The convolutions, whose cortical layers enable the cognitive functions, can be affected by various disorders. Some artists suffered from those disorders, and some others presented them in their artworks. CONCLUSIONS: The cerebral convolutions or gyri, thanks to their extensive cortical mantle, are the specific morphological basis for the human mind, but also the structures with their own aesthetics. Contemporary authors relatively often depictor model the cerebral convolutions, either from the aesthetic or conceptual aspect. In this way, they make a connection between the neuroscience and fineart.

Deposition of 137Cs and precipitation distribution in Vojvodina, Northern Serbia after the Chernobyl accident.

There is a dilemma whether the Chernobyl accident is the main source of this anthropogenic radionuclide in the region of Vojvodina, Northern Province of Serbia. The difference compared to the other dominant source of radiocaesium in the environment, the nuclear weapon tests, is lying in the local character of contamination due to accidents, which implies it's highly dependent on meteorological parameters, such as precipitation and wind in the area at that time. According to published data, the average value of surface contamination by radiocaesium on the territory of the former Yugoslavia after the Chernobyl accident was several times higher than the estimated contamination by nuclear testing (1945-1963). The main aim of this research is to explore possible correlations of the spatial distribution of precipitation from April 1986 to December 1987 with the deposition of radiocaesium in the surface soil to justify this claim in the absence of data on caesium deposition before the Chernobyl accident for the Vojvodina region. The database of 137Cs content in Vojvodina soil and precipitation for this region during the timeframe of interest were used. From a total of 245 precipitation stations in Vojvodina, 164 to 244 precipitation stations were selected for analysis in conditionals from different time scale aspects and data validation. The best correlation between amounts of total precipitation and 137Cs content in surface soil obtained for two rainy periods 2nd-5th and 7th-21st May 1986. Further cluster analysis separated four different regions according to 137Cs deposition and precipitation for the mentioned rainy period which could be applied in the field of soil erosion assessment on the local and regional level.

Percutaneous nephrostomy in the management of advanced and terminal-stage gynecologic malignancies: outcome and complications.

PURPOSE: The goal of the study was to evaluate the outcome and complications after percutaneous nephrostomy (PCN) insertion in advanced and terminal-stage gynecological malignancies with ureteral obstruction (UO). MATERIALS AND METHODS: We analyzed data of 117 patients with UO due to gynecological malignancies, who had undergone PCN between 1996 and 2006. Cervical cancer was evidenced in 108 patients, uterine carcinoma in six and ovarian cancer in three patients. Eighty-nine had UO at the initial manifestation of the disease, 22 had persistent or recurrent cancer, and six were disease-free after initial therapy. Oliguria was observed in 22.2% and creatine elevation in 79.5%. Mean follow-up was 11.43 months (range 0-112). RESULTS: The median age was 51 years (range 28-85). Bilateral nephrostomy was performed in 36.7% and unilateral in 63.3%. Renal function normalization occurred in 24.8%. Overall two-year survival (OS) was 16.8%. Higher OS occurred in patients without initial azotemia versus those with azotemia (26.8% vs 13.9%). Median survival time for all the patients was seven months, eight in primary cases versus six in recurrent ones, and eight months in patients after initial therapy. Complications appeared in 53.85%. Most frequent were the loss of the nephrostomy catheter in 37.61% and urinary tract infections in 19.6%. CONCLUSION: Improvement of renal function after PCN can be of clinical benefit in patients who might be cured or for prolonged palliative care. Azotemia seems to be poor prognostic sign.

A novel method for the functionalization of gamma-irradiated single wall carbon nanotubes with DNA.

In this work we describe a novel method for highly efficient functionalization of single wall carbon nanotubes (SWCNTs) by DNA wrapping. Exposure of SWCNTs to gamma-irradiation (50 kGy) has lowered by one order of magnitude the amount of single stranded deoxyribonucleic acid (ssDNA) required for SWCNT modification. The resulting hybrids of gamma-irradiated SWCNTs and ssDNA were characterized by optical absorbance spectroscopy, Raman spectroscopy and Fourier transform infrared spectroscopy. Atomic force microscopy was used to investigate the morphology of hybrids. While gamma-irradiation in three different media has significantly improved the process of SWCNT dispersion, irradiation in ammonia was the most efficient. The gamma-irradiated SWCNTs functionalized with ssDNA were stabilized by electrostatic forces. This preliminary study suggests that gamma-irradiation can significantly improve the functionalization of SWCNTs with DNA.

Comparative process analysis of fullerene production by the arc and the radio-frequency discharge methods.

In this work, comparative analysis of processes in carbon arc and radio frequency (RF) plasma during fullerene synthesis has been presented. The kinetic model of fullerene formation developed earlier has been verified in both types of plasma reactors. The fullerene yield depended on carbon concentration, velocity of plasma flame and rotational temperature of C2 radicals predominantly. When mean rotational temperature of C2 radicals was 3000 K, the fullerene yield was the highest regardless of the type of used reactor. The zone of fullerene formation is larger significantly in RF plasma reactor compared to arc reactor.

Loss of KAI1 messenger RNA expression in both high-grade and invasive human bladder cancers.

The molecular mechanisms responsible for metastasis are not fully understood. Recently, expression of the KAI1 gene on human chromosome 11p11.2 was found to be down-regulated in metastatic prostate cancer cell lines compared with normal human prostate, suggesting that KAI1 may be a metastasis suppressor gene. The aim of this study was to investigate whether there is reduced expression of KAI1 in late-stage bladder cancer. Sixty-six paraffin-embedded bladder tissue sections were analyzed for KAI1 mRNA by in situ hybridization. Nineteen of these were from patients with no histological evidence of bladder cancer, and 47 were from papillary transitional cell carcinomas (TCCs); of these, 16 were highly invasive. KAI1 mRNA was highly expressed in the specimens of normal bladder (11 of 11; 100%), inflammatory bladder (5 of 8; 63%), and noninvasive papillary TCCs of grades 1 and 2 (15 of 24; 63%), compared to grade 3 papillary TCCs (1 of 7; 14%) or invasive TCCs (1 of 16; 6%). The differences in expression between local and invasive disease were statistically significant (P

In vitro cytotoxicity of selected chemicals commonly produced during fire combustion using human cell lines.

Fire combustion products contain a broad range of chemicals, which have a multitude of possible toxic interactions in humans. The aim of this study was to evaluate the cytotoxicity of selected substances commonly produced during fire combustion. A range of human cell lines and cultures including: skin fibroblasts, HepG2 (liver derived), and A549 (lung derived cells) were used to represent different human target organs. The colorimetric MTS assay (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) was used to detect the cytotoxic effects of selected substances including: acetic acid, ammonia, formaldehyde, hydrobromic acid, hydrochloric acid, hydrofluoric acid, potassium cyanide, sodium fluoride, sodium nitrite, sodium sulphide, and sulphurous acid. In this study, the NOAEC (No Observable Adverse Effect Concentration), IC(10) (10% inhibitory concentration), IC50 (50% inhibitory concentration), and TLC (Total Lethal Concentration) values were determined. The ratio between in vitro IC50 to in vivo human toxicity data (Lowest Lethal Dose-LDLo and Lowest Lethal Concentration--LCLo) was also established. Results indicated a strong relationship between IC50 values on the cell types used: fibroblast and A549 (R2: 0.92), A549 and HepG2 (R2: 0.72), fibroblast and HepG2 (R2: 0.69). Good correlation was obtained between the IC50 against LDLo and LCLo when an appropriate adjustment factor was implemented. Results of this study indicated that in vitro methods could be a potential technique for assessing the toxicity of fire combustion products.

Nonbursal origin of humoral immunity: immune capacity and cytomorphological changes in chickens bursectomized as 52- to 64-hours-old embryos.

Chickens sham-bursectomized as 52- to 64-hours-old embryos (SBx), chickens bursectomized at hatching (NeoBx) and chickens from which the bursal primordium was removed at 52-64 h of embryonation (EBx) were immunized with guinea pig red blood cells when 21 days old. Following the second injection of antigen, EBx chickens were found to elaborate ME-sensitive and ME-resistant hemagglutinins, to produce direct and indirect plaque-forming cells, and to contain lymphocytes bearing a specific bursa (Bu) antigen. These immune properties were more often expressed in sham-bursectomized and neonatally bursectomized chickens. Cytomorphological changes in the spleen, bone marrow and cecal tonsil of EBx chickens were typical for birds lacking the bursa. The number of lymphocytes and plasma cells in the bone marrow and liver of SBx, NeoBx and EBx chickens was very low. These results suggest that a very early embryonic bursectomy of chickens does not abolish humoral immunity completely and does not prevent the differentiation of Bu antigen-bearing lymphocytes.

Quantitation of Fc gamma RII mRNA in platelets and megakaryoblastic cell lines by a new method of in situ hybridization.

We have developed a highly sensitive and quantitative, non-isotopic method of in situ hybridization in which the level of probe binding to intracellular mRNA is determined using an ELISA based detection method. Highly purified cell preparations or cells from a cultured cell line are centrifuged into 96 well microtiter plates. The cells are fixed with formalin and pre-treated with Triton X-100 and Nonidet P40 before photobiotin labeled cDNA probes are applied. The biotin from the hybridization is detected using multiple applications of streptavidin and biotinylated alkaline phosphatase and then visualized by the p-NPP (p-nitrophenyl phosphate) conversion method. We have determined a number of the optimal parameters in the procedure including the effects of cell numbers per well, development times and standardization of data using ubiquitous beta-actin mRNA and poly-A+ RNA expression as controls. We have used the technique to study the level of expression of FcgR mRNA in platelets and precursors. We found that platelets and megakaryoblastic cell lines only express mRNA for Fc gamma RII. The presence of the Fc gamma RII molecules was confirmed by complementary studies using immunohistochemistry with specific monoclonal antibodies IV.3 and KB61.

Preparation of specific anti-thymocyte and anti-bursacyte sera in rabbits.

A method of producing anti-thymocyte and anti-bursacyte sera in rabbits is described. Chickens which served as donors of cells were thoroughly perfused with saline to remove plasma proteins and circulating blood elements. For immunization, thymocytes were obtained from neonatally bursectomized birds, and bursacytes from chickens thymectomized at hatching. The purification of rabbit anti-lymphocyte sera included absorptions with leucocyte-free suspension of chicken erythrocytes, chicken liver cell membranes, thymocytes from bursectomized-irradiated and bursacytes from thymectomized-irradiated chickens, and chicken IgM and IgG immunoadsorbents. Cytotoxicity and fluorescent-antibody assays revealed that anti-thymocyte and anti-bursacyte sera thus produced clearly distinguished the surface antigenic determinants of thymocytes from those of bursacytes.

Suppression of experimental allergic encephalomyelitis in offspring of DA and Wistar rats following immunization of mother with encephalitogen.

Immunization of female rats with encephalitogen before gestation, during gestation, and during lactation differentially decreased susceptibility to experimental allergic encephalomyelitis (EAE) in their offspring. The most pronounced suppression, revealed by lowered incidence and weaker clinical signs of the disease, was observed in offspring of mothers immunized before gestation and during lactation in both Dark August (EAE-susceptible), and Wistar (EAE-relatively resistant) rat strains. Induction of EAE in mothers during pregnancy only delayed the onset of the disease in DA progeny. The overall effect on EAE in offspring did not depend on the disease intensity in mothers. Our results suggest that anti-myelin basic protein (MBP) antibodies passively transferred from mothers are not responsible for the observed protection in offspring.

Characterization of Fc gamma receptors on human megakaryocytes.

Megakaryocyte and platelet Fc gamma receptors (FcR) are of importance in the pathophysiology of immune complex-mediated thrombocytopenias such as heparin-induced thrombocytopenia. In this study, Fc gamma R proteins and mRNAs in normal human megakaryocytes were examined. Fc gamma R proteins were studied with immunocytochemical staining, dual colour flow cytometry and immunoprecipitation using monoclonal antibodies against Fc gamma R I, Fc gamma R II and Fc gamma R III. Fc gamma R mRNAs were measured with biotinylated cDNA of oligonucleotide probes using a novel quantitative in situ hybridization technique. Using these techniques, Fc gamma R II protein and mRNA, but not Fc gamma R I and Fc gamma R III proteins and transcripts were detected in megakaryocytes. Further, transcript analysis showed that megakaryocytes contain only the transcript of Fc gamma R IIA gene but no transcripts of Fc gamma R IIB nor Fc gamma R IIC genes; Fc gamma R IIA transcripts with and without the transmembrane (TM) exon are present in approximately equal proportions. In contrast, neutrophils and macrophages also contain Fc gamma R IIA transcript but Fc gamma R IIA transcript with the TM exon predominates suggesting cell lineage-specific Fc gamma R IIA expression. Fc gamma R IIA transcript lacking the TM exon predicts the presence of a potential soluble form of Fc gamma R in platelets and megakaryocytes which may have a physiological role as it can compete with the membrane-bound Fc gamma R IIA for binding of IgG-containing immune complexes and thus protect these cells from excessive binding and injurious effects of immune complexes.

The expression of cytokine messenger-rnas in clear cell acanthoma.

Clear cell acanthoma (syn: Degos' acanthoma) is a tumorous condition of the skin of unknown aetiology. It is seen as fleshy vascular papules which are solitary and are usually located on die lower extremities. We have studied 4 cases using cDNA-mRNA in situ hybridization for growth and immunological factors thought to be important molecular regulators in the skin. We obtained strongest binding with an interleukin-2 (IL-2) probe which bound to the tumour cells in the epidermis and to lymphoid cells in the dermal infiltrate. We also observed binding of probes for IL-1beta, IL-4, IL-6, IFN-alpha and TGF-alpha. Binding with the IL-2R probe was confined to scant cells in the tumour and a few cells in the lymphoid infiltrate. The significance of these observations and possible mechanisms for the pathogenesis of clear cell acanthoma are discussed.

Adsorption and dissolution behavior of human plasma fibronectin on thermally and chemically modified titanium dioxide particles.

Titanium is known for its biocompatibility and is widely used in dental and orthopedic reconstructive surgery. There are reports that osteointegration of these implants is not optimal. The objective of this study was to modify titanium dioxide particles and examine the resultant effects on protein adsorption to these altered surfaces using a model cell binding protein, human plasma fibronectin (HPF). HPF is an important matrix glycoprotein that plays a major role in cell and protein attachment, Titanium dioxide surfaces were modified by heating the titanium dioxide powder at 800 degrees C for 1 h or treating with an oxidizing agent: peroxide in ammonium hydroxide followed by peroxide in hydrochloric acid. Oxidized and control samples were further treated with 9:1 butanol:water for 30 min. Brunauer-Emmett-Teller showed no change in particle surface area as a result of thermal or chemical treatment. Hydrophobicity increased with butanol treatment of titanium dioxide. Diffuse reflectance Fourier transform infrared spectroscopy showed the presence of -CH2 and -CH3 vibrations in the region of 2850-3000 cm(-1) for both the heated, butanol and peroxide/butanol-treated samples. The absence of increased C-O and O-C=O features as determined by electron spectroscopy for chemical analysis indicates that butanol adsorption is not occurring via an esterification mechanism. The interaction between butanol and pre-heated or peroxide-treated titanium dioxide may be one of association (weak electrostatic and/or Van der Waals forces) rather than direct ionic bonding. Maximum HPF adsorption on modified or unmodified titanium dioxide occurred within 30 min, with greater protein adsorption occurring on butanol-treated samples. Desorption was minimal with all modifications. Zeta potential measurements showed that HPF adsorption caused an increase in the negative zeta potential with the greatest change noted for the butanol-treated samples. These findings suggest that wettability and surface charge both play an important role in protein adsorption to titanium dioxide. Thus, by modifying the physico-chemical properties of titanium dioxide surfaces, it may be possible to alter protein adsorption and hence optimize cell attachment.