Serum leptin levels in children and adolescents with spinal muscular atrophy types 2 and 3✰.

AIM: It has been reported that in patients with spinal muscular atrophy (SMA), lower levels of motor function are associated with hyperleptinemia. Additionally, hyperleptinemia has been found to be more frequent in underweight SMA patients. Therefore, we aimed to analyze serum leptin levels in patients with SMA. METHOD: This was a cross-sectional study of pediatric patients (2-19 years old) with SMA types 2 and 3. The investigations included anthropometric measurements, assessment of pubertal status, motor function evaluation using the Hammersmith Functional Motor Scale - Expanded (HFMSE), and serum leptin levels. RESULTS: In total, 37 patients (22 with type 2 and 15 with type 3 SMA) were included in the study. The male-to-female ratio was 1:1.3 and 62.2% of patients were prepubertal. No statistically significant correlation was found between the HFMSE score and leptin levels, rs(35) = 0.24, p = 0.15. There was, however, a strong positive relationship between the body mass index (BMI) z-score and leptin levels, rs(35) = 0.87, p < 0.001. CONCLUSION: Serum leptin levels do not seem to be a useful marker of disease severity in children and adolescents with types 2 and 3 SMA. As in the general pediatric population, leptin levels are strongly correlated with BMI, which is a surrogate measure of body fat.

Whole-genome analysis of extraintestinal Escherichia coli sequence type 73 from a single hospital over a 2 year period identified different circulating clonal groups.

Sequence type (ST)73 has emerged as one of the most frequently isolated extraintestinal pathogenic Escherichia coli. To examine the localized diversity of ST73 clonal groups, including their mobile genetic element profile, we sequenced the genomes of 16 multiple-drug resistant ST73 isolates from patients with urinary tract infection from a single hospital in Sydney, Australia, between 2009 and 2011. Genome sequences were used to generate a SNP-based phylogenetic tree to determine the relationship of these isolates in a global context with ST73 sequences (n=210) from public databases. There was no evidence of a dominant outbreak strain of ST73 in patients from this hospital, rather we identified at least eight separate groups, several of which reoccurred, over a 2 year period. The inferred phylogeny of all ST73 strains (n=226) including the ST73 clone D i2 reference genome shows high bootstrap support and clusters into four major groups that correlate with serotype. The Sydney ST73 strains carry a wide variety of virulence-associated genes, but the presence of iss, pic and several iron-acquisition operons was notable.

Assessing and visualising hazard impacts to enhance the resilience of Critical Infrastructures to urban flooding.

The design, construction and maintenance of Critical Infrastructures (CI) is commonly based on standards that are rigorous, so as to withstand any climate or weather-linked pressures. However, due to climate change, climate characteristics may shift, resulting in increased frequency/magnitude of potential failures, or exposure to new unknown risks. As vital components for the normal functioning of modern societies, the resilience of CIs under climate stressors encompasses their structural integrity, their operational elements, and their capacity to maximize business output. In this work, we propose an integrated and participatory methodological approach to enhance the resilience of interconnected CIs to urban flooding under climate change, by assessing the risk and introducing adaptation measures. The main objectives of the proposed methodology and approach are: (i) to provide scientific evidence for better understanding of how future climate regimes might affect normal operation of interconnected CI in urban areas during their lifespan; (ii) to assess the cost-effectiveness of different adaptation measures; (iii) to involve local stakeholders and operators in the co-design of the approach, as well as the assessment and the evaluation of adaptation measures; (iv) to combine computational modelling with advanced 3D visualisation techniques for effectively engaging stakeholders in decision making; (v) to include risk assessment and damage functions co-designed by end-users and local stakeholders; (vi) to integrate all of the aforementioned components in a specifically designed cloud platform as a Decision Support System for end-users, (vii) to validate the DSS by the end users and local stakeholders. The paper presents the computational background and tools. Additionally, it describes a Case Study in Torbay, UK, where the full methodology and the proposed participatory approach have been applied, with all the specifics, i.e., the scenarios of extreme flooding, the numerical and visualisation results, the response of the stakeholders and the evaluation of selected adaptation measures.

Mycoplasma hyopneumoniae resides intracellularly within porcine epithelial cells.

Enzootic pneumonia incurs major economic losses to pork production globally. The primary pathogen and causative agent, Mycoplasma hyopneumoniae, colonises ciliated epithelium and disrupts mucociliary function predisposing the upper respiratory tract to secondary pathogens. Alleviation of disease is reliant on antibiotics, vaccination, and sound animal husbandry, but none are effective at eliminating M. hyopneumoniae from large production systems. Sustainable pork production systems strive to lower reliance on antibiotics but lack of a detailed understanding of the pathobiology of M. hyopneumoniae has curtailed efforts to develop effective mitigation strategies. M. hyopneumoniae is considered an extracellular pathogen. Here we show that M. hyopneumoniae associates with integrin ß1 on the surface of epithelial cells via interactions with surface-bound fibronectin and initiates signalling events that stimulate pathogen uptake into clathrin-coated vesicles (CCVs) and caveosomes. These early events allow M. hyopneumoniae to exploit an intracellular lifestyle by commandeering the endosomal pathway. Specifically, we show: (i) using a modified gentamicin protection assay that approximately 8% of M. hyopneumoniae cells reside intracellularly; (ii) integrin ß1 expression specifically co-localises with the deposition of fibronectin precisely where M. hyopneumoniae cells assemble extracellularly; (iii) anti-integrin ß1 antibodies block entry of M. hyopneumoniae into porcine cells; and (iv) M. hyopneumoniae survives phagolysosomal fusion, and resides within recycling endosomes that are trafficked to the cell membrane. Our data creates a paradigm shift by challenging the long-held view that M. hyopneumoniae is a strict extracellular pathogen and calls for in vivo studies to determine if M. hyopneumoniae can traffic to extrapulmonary sites in commercially-reared pigs.

Towards satisfying performance of an O/W cosmetic emulsion: screening of reformulation factors on textural and rheological properties using general experimental design.

OBJECTIVE: Consumers' demand for improved products' performance, alongside with the obligation of meeting the safety and efficacy goals, presents a key reason for the reformulation, as well as a challenging task for formulators. Any change of the formulation, whether it is wanted - in order to innovate the product (new actives and raw materials) or necessary - due to, for example legislative changes (restriction of ingredients), ingredients market unavailability, new manufacturing equipment, may have a number of consequences, desired or otherwise. The aim of the study was to evaluate the influence of multiple factors - variations of the composition, manufacturing conditions and their interactions, on emulsion textural and rheological characteristics, applying the general experimental factorial design and, subsequently, to establish the approach that could replace, to some extent, certain expensive and time-consuming tests (e.g. certain sensory analysis), often required, partly or completely, after the reformulation. METHODS: An experimental design strategy was utilized to reveal the influence of reformulation factors (addition of new actives, preparation method change) on textural and rheological properties of cosmetic emulsions, especially those linked to certain sensorial attributes, and droplet size. RESULTS: The general experimental factorial design revealed a significant direct effect of each factor, as well as their interaction effects, on certain characteristics of the system and provided some valuable information necessary for fine-tuning reformulation conditions. Upon addition of STEM-liposomes, consistency, index of viscosity, firmness and cohesiveness were decreased, as along with certain rheology parameters (elastic and viscous modulus), whereas maximal and minimal apparent viscosities and droplet size were increased. The presence of an emollient (squalene) affected all the investigated parameters in a concentration-dependent manner. Modification of the preparation method (using Ultra Turrax instead of a propeller stirrer) produced emulsions with higher firmness and maximal apparent viscosity, but led to a decrease in minimal apparent viscosity, hysteresis loop area, all monitored parameters of oscillatory rheology and droplet size. CONCLUSION: The study showed that the established approach which combines a general experimental design and instrumental, rheological and textural measurements could be appropriate, more objective, repeatable and time and money-saving step towards developing cosmetic emulsions with satisfying, improved or unchanged, consumer-acceptable performance during the reformulation.

Marrubium vulgare ethanolic extract induces proliferation block, apoptosis, and cytoprotective autophagy in cancer cells in vitro.

Marrubium vulgare is a European medicinal plant with numerous beneficial effects on human health. The aim of the study was to isolate the plant ethanolic extract (MVE) and to investigate its anti-melanoma and anti-glioma effects. MVE was prepared by the modified pharmacopoeial percolation method and characterized by UHPLC-LTQ OrbiTrap MS. MVE dose-dependently reduced viability of melanoma (B16) and glioma (U251) cells, but not peripheral blood mononuclear cells. It arrested cell cycle in S+G2/M phase, which was associated with the activation of MAP kinase p38 and up-regulation of antiproliferative genes p53, p21 and p27. MVE induced oxidative stress, while antioxidants abrogated its antitumor effect. Furthermore, MVE induced mitochondrial depolarization, activation of caspase-9 and -3, Parp cleavage, phosphatidylserine exposure and DNA fragmentation. The mitochondrial apoptotic pathway was associated with the up-regulation of proapoptotic genes Pten, Bak1, Apaf1, and Puma and down-regulation of antiapoptotic genes survivin and Xiap. MVE also stimulated the expression of autophagy-related genes Atg5, Atg7, Atg12, Beclin-1, Gabarab and Sqstm1, as well as LC3-I conversion to the autophagosome associated LC3-II, while autophagy inhibitors exacerbated its cytotoxicity. Finally, the most abundant phenolic components of MVE, ferulic, p-hydroxybenzoic, caffeic and chlorogenic acids, did not exert a profound effect on viability of tumor cells, suggesting that other components individually or in concert are the mediators of the extracts' cytotoxicity. By demonstrating the ability of MVE to inhibit proliferation, induce apoptosis and cytoprotective autophagy, our results suggest that MVE, alone or combined with autophagy inhibitors, could be a good candidate for anti-melanoma and anti-glioma therapy.

Temporal dynamics and subpopulation analysis of Theileria orientalis genotypes in cattle.

In Australia, outbreaks of clinical theileriosis caused by Theileria orientalis have been largely associated with the Ikeda genotype which can occur as a sole infection, or more commonly, as a mixture of genotypes. The most prevalent genotype, Chitose, frequently co-occurs with type Ikeda, however the role of this genotype in clinical disease has not been clearly established. Furthermore, the dynamics of individual genotypes in field infection of cattle have not been examined. In this study we developed quantitative PCR (qPCR) and genotyping methods to examine the role of the Chitose genotype in clinical disease and to investigate the temporal dynamics of T. orientalis Ikeda, Chitose and Buffeli genotypes in naïve animals introduced to a T. orientalis-endemic area. Analysis of the major piroplasm surface protein (MPSP) genes of Chitose isolates revealed the presence of two distinct phylogenetic clusters, Chitose A and Chitose B. A genotyping assay aimed at determining Chitose A/B allele frequency revealed that the Chitose A phylogenetic cluster is strongly associated with clinical disease but nearly always co-occurs with the Ikeda genotype. qPCR revealed that the Chitose genotype (particularly Chitose A), undergoes temporal switching in conjunction with the Ikeda genotype and contributes substantially to the overall parasite burden. The benign Buffeli genotype can also undergo temporal switching but levels of this genotype appear to remain low relative to the Ikeda and Chitose types. Interplay between vector and host immunological factors is presumed to be critical to the population dynamics observed in this study. Genotypic switching likely contributes to the persistence of T. orientalis in the host.

A low-power, high-speed, 9-channel germanium-silicon electro-absorption modulator array integrated with digital CMOS driver and wavelength multiplexer.

We demonstrate the first germanium-silicon C-band electro-absorption based waveguide modulator array and echelle-grating-based silicon wavelength multiplexer integrated with a digital CMOS driver circuit. A 9-channel, 10Gbps SiGe electro-absorption wavelength-multiplexed modulator array consumed a power of 5.8mW per channel while being modulated at 10.25Gbps by 40nm CMOS drivers delivering peak-to-peak voltage swings of 2V, achieving a modulation energy-efficiency of ~570fJ/bit including drivers. Performance up to 25Gbps on a single-channel SiGe modulator and CMOS driver is also reported.

Decision making in flood risk based storm sewer network design.

It is widely recognised that flood risk needs to be taken into account when designing a storm sewer network. Flood risk is generally a combination of flood consequences and flood probabilities. This paper aims to explore the decision making in flood risk based storm sewer network design. A multiobjective optimization is proposed to find the Pareto front of optimal designs in terms of low construction cost and low flood risk. The decision making process then follows this multi-objective optimization to select a best design from the Pareto front. The traditional way of designing a storm sewer system based on a predefined design storm is used as one of the decision making criteria. Additionally, three commonly used risk based criteria, i.e., the expected flood risk based criterion, the Hurwicz criterion and the stochastic dominance based criterion, are investigated and applied in this paper. Different decisions are made according to different criteria as a result of different concerns represented by the criteria. The proposed procedure is applied to a simple storm sewer network design to demonstrate its effectiveness and the different criteria are compared.

Thermal and ultrasonic influence in the formation of nanometer scale hydroxyapatite bio-ceramic.

Hydroxyapatite (HAP) is a widely used biocompatible ceramic in many biomedical applications and devices. Currently nanometer-scale forms of HAP are being intensely investigated due to their close similarity to the inorganic mineral component of the natural bone matrix. In this study nano-HAP was prepared via a wet precipitation method using Ca(NO(3))(2) and KH(2)PO(4) as the main reactants and NH(4)OH as the precipitator under ultrasonic irradiation. The Ca/P ratio was set at 1.67 and the pH was maintained at 9 during the synthesis process. The influence of the thermal treatment was investigated by using two thermal treatment processes to produce ultrafine nano-HAP powders. In the first heat treatment, a conventional radiant tube furnace was used to produce nano-particles with an average size of approximately 30 nm in diameter, while the second thermal treatment used a microwave-based technique to produce particles with an average diameter of 36 nm. The crystalline structure and morphology of all nanoparticle powders produced were investigated using X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), and Fourier transform infrared spectroscopy (FT-IR). Both thermal techniques effectively produced ultrafine powders with similar crystalline structure, morphology and particle sizes.

Calibration of a 1D/1D urban flood model using 1D/2D model results in the absence of field data.

Recently increased flood events have been prompting researchers to improve existing coupled flood-models such as one-dimensional (1D)/1D and 1D/two-dimensional (2D) models. While 1D/1D models simulate sewer and surface networks using a one-dimensional approach, 1D/2D models represent the surface network by a two-dimensional surface grid. However their application raises two issues to urban flood modellers: (1) stormwater systems planning/emergency or risk analysis demands for fast models, and the 1D/2D computational time is prohibitive, (2) and the recognized lack of field data (e.g. Hunter et al. (2008)) causes difficulties for the calibration/validation of 1D/1D models. In this paper we propose to overcome these issues by calibrating a 1D/1D model with the results of a 1D/2D model. The flood-inundation results show that: (1) 1D/2D results can be used to calibrate faster 1D/1D models, (2) the 1D/1D model is able to map the 1D/2D flood maximum extent well, and the flooding limits satisfactorily in each time-step, (3) the 1D/1D model major differences are the instantaneous flow propagation and overestimation of the flood-depths within surface-ponds, (4) the agreement in the volume surcharged by both models is a necessary condition for the 1D surface-network validation and (5) the agreement of the manholes discharge shapes measures the fitness of the calibrated 1D surface-network.

An analysis of the combined consequences of pluvial and fluvial flooding.

Intense rainfall in urban areas often generates both pluvial flooding due to the limited capacity of drainage systems, as well as fluvial flooding caused by deluges from river channels. The concurrence of pluvial and fluvial flooding can aggravate their (individual) potential damages. To analyse the impact caused by individual and composite type of flooding, the SIPSON/UIM model, an integrated 1D sewer and 2D overland flow was applied to numerical modelling. An event matrix of possible pluvial scenarios was combined with hypothetic overtopping and breaching situations to estimate the surface flooding consequences in the Stockbridge area, Keighley (Bradford, UK). The modelling results identified different flooding drivers in different parts of the study area and showed that the worst scenarios resulted from synthesised events.

Identification of lipoprotein MslA as a neoteric virulence factor of Mycoplasma gallisepticum.

Many lipoproteins are expressed on the surfaces of mycoplasmas, and some have been implicated as playing roles in pathogenesis. Family 2 lipoproteins of Mycoplasma pneumoniae have a conserved "mycoplasma lipoprotein X" central domain and a "mycoplasma lipoprotein 10" C-terminal domain and are differentially expressed in response to environmental conditions. Homologues of family 2 lipoproteins are Mycoplasma specific and include the lipoprotein of Mycoplasma gallisepticum, encoded by the MGA0674 gene. Comparative transcriptomic analysis of the M. gallisepticum live attenuated vaccine strain F and the virulent strain R(low), reported in this study, indicated that MGA0674 is one of several differentially expressed genes. The MGA0674-encoded lipoprotein is a proteolytically processed, immunogenic, TX-114 detergent-phase protein which appears to have antigenic divergence between field strains R(low) and S6. We examined the virulence of an R(low) Delta MGA0674 mutant (P1H9) in vivo and observed reduced recovery and attenuated virulence in the tracheas of experimentally infected chickens. The virulence of two additional R(low) Delta MGA0674 mutants, 2162 and 2204, was assessed in a second in vivo virulence experiment. These mutants exhibited partial to complete attenuation in vivo, but recovery was observed more frequently. Since only Mycoplasma species harbor homologues of MGA0674, the gene product has been renamed "Mycoplasma-specific lipoprotein A" (MslA). Collectively, these data indicate that MslA is an immunogenic lipoprotein exhibiting reduced expression in an attenuated strain and plays a role in M. gallisepticum virulence.

Mycoplasma leachii sp. nov. as a new species designation for Mycoplasma sp. bovine group 7 of Leach, and reclassification of Mycoplasma mycoides subsp. mycoides LC as a serovar of Mycoplasma mycoides subsp. capri.

The Mycoplasma mycoides cluster consists of six pathogenic mycoplasmas causing disease in ruminants, which share many genotypic and phenotypic traits. The M. mycoides cluster comprises five recognized taxa: Mycoplasma mycoides subsp. mycoides Small Colony (MmmSC), M. mycoides subsp. mycoides Large Colony (MmmLC), M. mycoides subsp. capri (Mmc), Mycoplasma capricolum subsp. capricolum (Mcc) and M. capricolum subsp. capripneumoniae (Mccp). The group of strains known as Mycoplasma sp. bovine group 7 of Leach (MBG7) has remained unassigned, due to conflicting data obtained by different classification methods. In the present paper, all available data, including recent phylogenetic analyses, have been reviewed, resulting in a proposal for an emended taxonomy of this cluster: (i) the MBG7 strains, although related phylogenetically to M. capricolum, hold sufficient characteristic traits to be assigned as a separate species, i.e. Mycoplasma leachii sp. nov. (type strain, PG50(T) = N29(T) = NCTC 10133(T) = DSM 21131(T)); (ii) MmmLC and Mmc, which can only be distinguished by serological methods and are related more distantly to MmmSC, should be combined into a single subspecies, i.e. Mycoplasma mycoides subsp. capri, leaving M. mycoides subsp. mycoides (MmmSC) as the exclusive designation for the agent of contagious bovine pleuropneumonia. A taxonomic description of M. leachii sp. nov. and emended descriptions of M. mycoides subsp. mycoides and M. mycoides subsp. capri are presented. As a result of these emendments, the M. mycoides cluster will hereafter be composed of five taxa comprising three subclusters, which correspond to the M. mycoides subspecies, the M. capricolum subspecies and the novel species M. leachii.

Characterisation of Erysipelothrix rhusiopathiae isolates from pigs associated with vaccine breakdowns.

Swine erysipelas vaccines are routinely used to protect pigs against peracute and acute/urticarial forms of Erysipelothrix. Between 1995 and 1998, 34 swine herds across four Australian states experienced vaccine failure. Forty-four isolates of Erysipelothrix rhusiopathiae of serovars 2, 1a, 1b and 1bx21 were recovered from 15 of these 34 vaccine breakdown herds. These isolates were characterised by restriction fragment length polymorphism (RFLP) analyses using RsaI and AluI on whole cell DNA and for the presence of plasmid DNA. Results were compared with those of 20 isolates from 16 herds unaffected by vaccine breakdown and 13 isolates representing 10 reference strains. The majority of breakdown herds possessed isolates of serovar 2 (9/15 herds), followed by serovar 1a (5 herds). No geographic predominance of a single serovar was evident. The identification of 10 RsaI profiles from whole cell DNA among the 44 isolates from 15 breakdown herds indicated that a single, new clonal lineage of E. rhusiopathiae was not responsible for vaccine failure. RsaI RFLP analyses detected a further 14 distinct profiles among 20 field strains unassociated with vaccine breakdowns, and none matched profiles of the 10 serovar reference strains for serovars 1a, 1b, 2 or 21. This technique is recommended for epidemiological studies of E. rhusiopathiae strains.

Automatic calibration of urban drainage model using a novel multi-objective genetic algorithm.

In order to successfully calibrate an urban drainage model, multiple calibration criteria should be considered. This raises the issue of adopting a method for comparing different solutions (parameter sets) according to a set of objectives. Amongst the global optimization techniques that have blossomed in recent years, Multi Objective Genetic Algorithms (MOGA) have proved effective in numerous engineering applications, including sewer network modelling. Most of the techniques rely on the condition of Pareto efficiency to compare different solutions. However, as the number of criteria increases, the ratio of Pareto optimal to feasible solutions increases as well. The pitfalls are twofold: the efficiency of the genetic algorithm search worsens and decision makers are presented with an overwhelming number of equally optimal solutions. This paper proposes a new MOGA, the Preference Ordering Genetic Algorithm, which alleviates the drawbacks of conventional Pareto-based methods. The efficacy of the algorithm is demonstrated on the calibration of a physically-based, distributed sewer network model and the results are compared with those obtained by NSGA-II, a widely used MOGA.

SIPSON--simulation of interaction between pipe flow and surface overland flow in networks.

The new simulation model, named SIPSON, based on the Preissmann finite difference method and the conjugate gradient method, is presented in the paper. This model simulates conditions when the hydraulic capacity of a sewer system is exceeded, pipe flow is pressurized, the water flows out from the piped system to the streets, and the inlets cannot capture all the runoff. In the mathematical model, buried structures and pipelines, together with surface channels, make a horizontally and vertically looped network involving a complex interaction of flows. In this paper, special internal boundary conditions related to equivalent inlets are discussed. Procedures are described for the simulation of manhole cover loss, basement flooding, the representation of street geometry, and the distribution of runoff hydrographs between surface and underground networks. All these procedures are built into the simulation model. Relevant issues are illustrated on a set of examples, focusing on specific parameters and comparison with field measurements of flooding of the Motilal ki Chal catchment (Indore, India). Satisfactory agreement of observed and simulated hydrographs and maximum surface flooding levels is obtained. It is concluded that the presented approach is an improvement compared to the standard "virtual reservoir" approach commonly applied in most of the models.

Liquid chromatographic-mass spectrometric method for the determination of fluoxetine and norfluoxetine in human plasma: application to clinical study.

A selective, sensitive, simple, and rapid method for the simultaneous determination of fluoxetine (FL) and norfluoxetine (nor-FL) was developed and validated, and further applied to analyze plasma samples obtained from FL-treated patients with Parkinson disease (n = 18). After one step liquid-liquid extraction with ethyl acetate, plasma samples were chromatographed on a C8 column. The mobile phase was acetate buffer and acetonitrile (40:60 v/v). Determination of FL and nor-FL was performed with MS detection in selective ion monitoring (SIM) mode, so the other components did not interfere with this assay. FL, nor-FL and flumazenil as internal standard were eluted in 6 min. Recoveries ranged from 89.7 to 96.6% and from 80.2 to 85.3% for FL and nor-FL, respectively. The limit of quantitation under described conditions was 2.5 microg/l for FL and 10 microg/l for nor-FL. The method was found to be reproducible with coefficient of variation less than 9%. The parameters of the method were found to be acceptable to enable its routine use for clinical studies. The method was employed to analyze the Parkinsonian patients' plasma samples. A great deviation in plasma concentrations of FL and nor-FL found among 18 studied patients indicates high pharmacokinetic variability of the drug. Obtained results also indicate absence of the influence of Parkinson disease on the drug disposition.

Plasma albumin level as an indicator of severity of preeclampsia.

We propose that hypoalbuminemia in preeclampsia is the result of reduced hepatic blood flow which is secondary to hypovolemia created by higher filtration pressure in the capillaries. Thus, hypoalbuminemia can be identified as an early sign in developing preeclampsia. We reviewed the medical records of 60 patients, aged 22-28, admitted with a diagnosis of preeclampsia during the third trimester of pregnancy. Normotensive patients served as a control group. Albumin levels were correlated with severity of the disorder. Levels between 3.0-3.5 g/dl were seen in both groups. All patients with severe preeclampsia had values below 3.0 gm/dl. Serum albumin levels may serve as an indicator of the severity of preeclampsia.

The separation of hypericine and pseudohypericine from Hypericum perforatum L.

The extraction of Hypericum perforatum L. was carried out by the method of ultrasonic maceration. The separation of H. perforatum L. extract in order to obtain hypericine and pseudohypericine rich fractions was carried out by the following chromatographic methods: flash column chromatography, high speed countercurrent chromatography, XAD solid phase extraction and Sephadex column chromatography. The separation by the Sephadex column chromatography gave the best results. Preparative HPLC was used to isolate hypericine and pseudohypericine from fraction 4 obtained by the Sephadex column chromatography procedure.