Association of methylenetetrahydrofolate (MTHFR) and apolipoprotein E (apo E) genotypes with homocysteine, vitamin and lipid levels in carotid stenosis

The aim of the study was to investigate the association between methylenetetrahydrofolate (MTHFR) genotypes and levels of homocysteine (Hcy), folate, vitamin B12 and lipids as well as the association between apolipoprotein E (apo E) genotypes and levels of lipids in a Croatian healthy control group and a group of patients with > 70% carotid stenosis (CS). The study included 98 Croats, 38 patients with > 70% carotid stenosis and 60 age- and sex-matched controls. The MTHFR and apo E genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), Hcy by enzyme immunoassay, vitamins by immunochemiluminiscence, and lipids by spectrophotometric method. There was no difference between control subjects and CS patients in the distribution of C677T MTHFR genotypes (p=0. 786) and alleles (p=0.904), however, differences in the frequencies of apo E genotypes (p=0.012) and alleles (p=0.029) were statistically significant. The odds ratio for apo E 3/4 genotype was 3.93 (95% CI 1.23-12.61). Hyperhomocysteinemia (> or =15 micromol/L) was found in 11% of CS patients and 5% of control subjects. Total cholesterol, triglycerides, vitamin B12 and folate were statistically different in "all MTHFR genotypes" (p<0.001, p<0.01, p=0.044 and p=0.036, respectively), and in TC/TT (p<0.001, p=0.003, p=0.030 and p=0.032, respectively) groups. The levels of total cholesterol, LDL cholesterol and triglycerides in the apo E 3/3, and total cholesterol in the apo E 3/4 group yielded statistical difference. An association was found of apo E 3/4 genotype but not of MTHFR genotypes with the risk of CS. MTHFR and apo E affect blood lipid levels, which was statistically confirmed. An association was also recorded between hyperhomocysteinemia and patients with CS. Vitamin status in CS showed a statistically verified association with TC/TT MTHFR genotype. In the group of patients with TC/TT MTHFR genotype, lower vitamin B12 and higher folate values were recorded. The results of multiple logistic analysis showed that there was no statistical significance of Hcy levels (OR 2.403, p=0.334) or conventional vascular risk factors such as smoking habit (OR 0.505, p=0.149), age (OR 1.048, p=0.087) or sex (OR 2.037, p=0.112) in predicting CS.     

Do we need the "adolescent crisis" diagnosis?

The aim of the study was to examine patients in adolescent crisis at the beginning of treatment and after a period of 12 months in order to evaluate the relative diagnostic and therapeutic validity. The study included 153 Split University students in adolescent crisis; 90 of them were treated by counseling and 63 served as controls. For diagnosis, Hampstead index and Diagnostic and Statistical Manual of Mental Disorders Fourth Edition (DSM-IV) multiaxial evaluation were used, allowing a wider insight into personal functioning. The study sample was split in 7 significantly different diagnostic subgroups. The counseling-treated examinees had better personality functioning after 12 months, but did not differ significantly from the control group. Some of their single functions were more severely disturbed at the very beginning. Counseling is a valuable therapeutic and diagnostic tool for adolescent crisis. The assessment must evaluate the entire person, because looking at only one aspect, due to different development and its place, a wrong conclusion may be reached. The "adolescents crisis" entity is clinically relevant.     

Can we detect the development of baroreflex sensitivity in humans between 11 and 20 years of age?

The aim of the study was to determine changes of baroreflex sensitivity in humans between 11 and 20 years of age. Continuous 5 min blood pressure recordings using a Finapres were taken in 415 healthy subjects while in a sitting, resting position (breathing at a frequency of 0.33 Hz). Beat-by-beat values of interbeat intervals (IBI) or heart rate, and systolic and diastolic blood pressures were measured. Baroreflex sensitivity in ms/mmHg (BRS) and in mHz/mmHg (BRSf) was determined at an average frequency of 0.1 Hz by spectral analysis. BRS did not correlate with age, but BRSf significantly decreased with age (p < 0.001). BRS correlated with mean IBI (p < 0.001) in all subjects and also in the particular subgroups, but BRSf was IBI-independent. Results of multiregression equations were BRS = 1.37 - 0.56 x age (years) + 0.02 x IBI (ms) (p < 0.001 for BRS vs. age and for BRS vs. IBI); BRSf = 34.74 - 0.97 x age (years) - 0.001 x IBI (ms) (p < 0.001 only for BRS vs. age), where age was measured in years and IBI was measured in ms. The limits of BRS were estimated for the total group: 5th percentile, 3.9; 50th percentile, 9.1; and 95th percentile, 18.7 ms/mmHg; and limits for BRSf were 5th percentile, 8.5; 50th percentile, 16.4; and 95th percentile, 33.6 mHz/mmHg. We conclude that IBI-dependent BRS was unchanged in the particular age groups, but the standardization of BRS on IBI decreased with age. BRSf was IBI-independent and better reflected the development of the BRS.     

Anomalous scattering analysis of Agrobacterium radiobacter phosphotriesterase: the prominent role of iron in the heterobinuclear active site

Bacterial phosphotriesterases are binuclear metalloproteins for which the catalytic mechanism has been studied with a variety of techniques, principally using active sites reconstituted in vitro from apoenzymes. Here, atomic absorption spectroscopy and anomalous X-ray scattering have been used to determine the identity of the metals incorporated into the active site in vivo. We have recombinantly expressed the phosphotriesterase from Agrobacterium radiobacter (OpdA) in Escherichia coli grown in medium supplemented with 1 mM CoCl2 and in unsupplemented medium. Anomalous scattering data, collected from a single crystal at the Fe-K, Co-K and Zn-K edges, indicate that iron and cobalt are the primary constituents of the two metal-binding sites in the catalytic centre (alpha and beta) in the protein expressed in E. coli grown in supplemented medium. Comparison with OpdA expressed in unsupplemented medium demonstrates that the cobalt present in the supplemented medium replaced zinc at the beta-position of the active site, which results in an increase in the catalytic efficiency of the enzyme. These results suggest an essential role for iron in the catalytic mechanism of bacterial phosphotriesterases, and that these phosphotriesterases are natively heterobinuclear iron-zinc enzymes.     

Conformational stability of 17 beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus

The functional activities of proteins are closely related to their molecular structure and understanding their structure-function relationships remains one of the intriguing problems of molecular biology. We investigated structural changes in 17beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus (17beta-HSDcl) induced by pH, temperature, salt, urea, guanidine hydrochloride, and coenzyme NADPH binding. At 25 degrees C and within the relatively narrow pH range of 7.0-9.0, 17beta-HSDcl exists in its native conformation as a dimer. This native conformation is thermally stable up to 40 degrees C in this pH range. At 25 degrees C and pH 2.0 in the presence of 150-300 mM NaCl, 17beta-HSDcl forms soluble aggregates enriched in alpha-helical and beta-sheet structures. At higher temperatures and NaCl concentrations, these soluble aggregates start to precipitate. The denaturants urea and guanidine hydrochloride unfold 17beta-HSDcl at concentrations of 1.2 and 0.4 M, respectively. Binding of the coenzyme NADPH to 17beta-HSDcl causes local structural changes that do not significantly affect the thermal stability of this protein.     

Structure and thermal behavior of the layered zincophosphate [NH3-CH2-CH(NH3)-CH3](ZnPO4)2

A zinc phosphate with the composition [NH₃-CH₂-CH(NH₃)-CH₃](ZnPO₄)₂, containing the doubly protonated 1,2-diaminopropane (abbr. HDAP), was synthesized by hydrothermal crystallization of zinc nitrate, phosphoric acid, 1,2-diaminopropane and trimethylenedipiperidine. The single crystal analysis shows a layered inorganic-organic structure built up of a sandwich-like motif of alternating inorganic layers and HDAP cations (trimethylenedipiperidine not being a constituent of the structure). The HDAP species are found to be disordered and serve as bridges between two adjacent inorganic layers which are separated by 3.95 Å. The bridging interaction occurs via a hydrogen-bonding network. The inorganic layer features a pattern of four-membered rings involving two ZnO₄ and two PO₄ tetrahedra connected by sharing O atoms. Thermal analysis shows that the compound is stable up to 370 °C and that the thermal decomposition of HDAP occurs in two steps between 370 and 460 °C, causing the collapse of the structure. The first decomposition step corresponds to ammonia removal which proceeds with a high activation energy (E_a = 282 kJ mol⁻¹). The high E_a value is mainly attributed to strong electrostatic interactions between organic cations and anionic inorganic layers, the disruption of the interactions being the main reason for structural collapse after the HDAP removal.     

HIV risks among female sex workers in Croatia and Montenegro

The study analyzed the prevalence and determinants of HIV-risks among female sex workers (FSWs) in Croatia and Montenegro. Face-to-face interviews were carried out in Zagreb, Split, and Podgorica during the 2006-2008 period. Croatian participants (n = 154) reported fewer clients, more consistent condom use, higher rates of HIV testing, and greater HIV knowledge. The participants interviewed in Montenegro (n = 119) were more likely to have injected drugs and to have experienced sexual abuse in the previous year. Although Montenegrin FSWs were more exposed to HIV-risks than Croatian FSWs, they reported lower HIV-risk self-assessment. Consistent condom use was significantly associated with education and HIV-risk self-assessment in the Croatian and the experience of physical/sexual abuse in the Montenegrin sample. In spite of a number of methodological limitations, the empirical insights provided by this study may assist in improving the existing HIVV prevention initiatives.     

Psychophysical characteristics of the premenstrual period

Aim of this study was to investigate psychophysical characteristics of the premenstrual period in regard to the period following menstruation. Psychophysical characteristics are defined as assessments of the physical state (fatigue, breast tenderness, bloating and lack of sleep), emotional state (happiness, anger, sadness, irritability, anxiety and distraction) and general assessments of physical attractiveness and satisfaction with oneself Diary of psychophysical characteristics was designed for this study as well as a socio-demographic questionnaire and a menstrual cycle calendar. 51 healthy, non-help-seeking women, aged 21-46 kept a diary of psychophysical characteristics a week before menstruation and four days one week after the onset of menstruation, during two menstrual cycles. The premenstrual period is characterized by the intensification of negative emotional states, the decline of positive emotional states, and the worsening of the general self-image through the decline of the perception of physical attractiveness and satisfaction with oneself In conclusion, the premenstrual period is not marked only by the mood-swings but also by the changes in the cognitive representation of self     

Effect of Encapsulated Probiotic Starter Culture on Rheological and Structural Properties of Natural Hydrogel Carriers Affected by Fermentation and Gastrointestinal Conditions

The suitability of natural hydrogel carriers with probiotic starter culture as whey beverages supplements was examined by assessing their rheological and structural changes during the fermentation and gastrointestinal conditions. Effect of encapsulated cells on the carrier structure is of great importance for the selection of proper material for the preparation of functional supplements. The structural changes of the chitosan-coated alginate/whey carriers were considered based on (1) cell viability and the carrier average volume vs. time (2) the storage and loss modulus vs. time obtained under low oscillator strain conditions, (3) FTIR analysis and (4) SEM cross-sectional observation of the hydrogel carriers. The presence of chitosan coating and fermentation conditions increased cell viability up to 9.01 ± 0.18 (log CFU/g). According to our results, the encapsulated cells induce weakening of carriers under the gastric conditions but improve their mechanical stability under the intestinal condition. The mechanical behaviour of carriers was also considered in order to formulate the rheological constitutive model equation for describing the irreversible structural changes under the gastric and intestinal conditions. The cell leakage under the gastric condition after the 2 h was less than 5%. Carriers are rapidly degraded under the intestinal condition which ensures the release of cells and provides their beneficial effects on the host health. Our results indicate that this type of coated carrier is suitable to be used for encapsulation of probiotic starter culture in the production of fermented whey-based products.

     

ERRγ enhances UCP1 expression and fatty acid oxidation in brown adipocytes

Objective:

Estrogen‐related receptors (ERRs) are important regulators of energy metabolism. Here we investigated the hypothesis that ERRγ impacts on differentiation and function of brown adipocytes.

Design and Methods:

We characterize the expression of ERRγ in adipose tissues and cell models and investigate the effects of modulating ERR? activity on UCP1 gene expression and metabolic features of brown and white adipocytes.

Results:

ERRγ was preferentially expressed in brown compared to white fat depots, and ERRγ was induced during cold‐induced browning of subcutaneous white adipose tissue and brown adipogenesis. Overexpression of ERRγ positively regulated uncoupling protein 1 (UCP1) expression levels during brown adipogenesis. This ERRγ‐induced augmentation of UCP1 expression was independent of the presence of peroxisome proliferator‐activated receptor coactivator‐1 (PGC‐1α) but was associated with increased rates of fatty acid oxidation in adrenergically stimulated cells. ERR? did not influence mitochondrial biogenesis, and its reduced expression in white adipocytes could not explain their low expression level of UCP1.

Conclusions:

Through its augmenting effect on expression of UCP1, ERRγ may physiologically be involved in increasing the potential for energy expenditure in brown adipocytes, a function that is becoming of therapeutic interest.