HCV genotype shift occurred over the 15 years in PWIDs in the Czech Republic.

OBJECTIVES: To follow on the epidemiology of HCV, especially genotypes spreading among people who inject drugs (PWID) in Prague and surrounding Central Bohemia, Czech Republic. METHODS: 546 patients who reported past and/or recent injecting of drugs were recruited in the years 2010-2012. They were initially tested for anti-HCV. Real-time PCR was used for quantification and genotyping of hepatitis C virus. Obtained data from the years 2010-2012 were compared with historical controls from periods of 1998-2000 and 2005-2007. RESULTS: Of 546 initially recruited and tested patients were 393 (72%) anti-HCV seropositive and of them 269 (68.4%) had detectable HCV PCR RNA. The most prevalent subtype was 3a in 97 patients (36.1%), 1a was detected in 85 patients (31.6%) and 1b in 57 patients (21.2%). These three genotypes were responsible for nearly 89% of infections. CONCLUSION: Significant increase in both genotypes 1a and 3a over the 15 years was apparent and significant, followed by the decrease in genotype 1b. In the genotype 1b and genotype 3a the significance has risen with the years of data collection. Described genotypic shift reflects the evolution of HCV epidemics and corresponds with the mode of transmission.

Electrocatalytic monitoring of peptidic proton-wires.

The transfer of protons or proton donor/acceptor abilities is an important phenomenon in many biomolecular systems. One example is the recently proposed peptidic proton-wires (H-wires), but the ability of these His-containing peptides to transfer protons has only been studied at the theoretical level so far. Here, for the first time the proton transfer ability of peptidic H-wires is examined experimentally in an adsorbed state using an approach based on a label-free electrocatalytic reaction. The experimental findings are complemented by theoretical calculations at the ab initio level in a vacuum and in an implicit solvent. Experimental and theoretical results indicated Ala3(His-Ala2)6 to be a high proton-affinity peptidic H-wire model. The methodology presented here could be used for the further investigation of the proton-exchange chemistry of other biologically or technologically important macromolecules.

The Endocannabinoid System - The Prediction of Spontaneous Preterm Birth in High-Risk Women: Protocol of a Study.

Spontaneous preterm birth (sPTB) is a major cause of perinatal morbidity and mortality, even in developed countries. Prediction of sPTB is therefore a valuable tool to reduce the associated risks. The current standard for the prediction of sPTB consists, in addition to anamnestic data, of previous sPTB and previous second trimester miscarriage, measurement of cervical length by transvaginal ultrasound (TVU CL) together with assessment of fetal fibronectin levels in cervicovaginal fluid. Other evaluation parameters, such as the level of endocannabinoids in the pregnant woman's blood, could increase the sensitivity of this management. Endocannabinoids (eCBs) are a part of the endocannabinoid system (ECS); out of them anandamide (arachidonoyl-ethanolamide, AEA), in particular, plays an important role in the regulation of pregnancy and childbirth. We present the protocol for an open, non-randomized study to evaluate concentrations of AEA and other endocannabinoids: 2 linoleoylglycerol (2-AG), 2 linoleoylglycerol (2-LG), 2 oleoylglycerol (2-OG), and 2 arachidonoyldopamine (2-ADOPA or also NADA) in the blood of pregnant women as potential predictors of sPTB. In a total of 230 women with a history of sPTB or miscarriage, eCBs levels between 22 and 28 weeks of gestation will be assessed from maternal blood, in addition to the standard procedure. The aim of the study is to determine the relationship between blood concentrations of the endocannabinoids tested and the risk of sPTB. The results of this study will describe the prognostic significance of maternal blood eCBs levels for sPTB, and could subsequently enable improved screening programs for early identification of sPTB.

Solid-phase/supercritical-fluid extraction for liquid chromatography of phenolic compounds in freshwater microalgae and selected cyanobacterial species.

In the present paper a new extraction technique based on the combination of solid-phase/supercritical-fluid extraction (SPE/SFE) with subsequent reversed-phase HPLC is described. The SPE/SFE extractor was originally constructed from SPE-cartridge incorporated into the SFE extraction cell. Selected groups of benzoic acid derivatives (p-hydroxybenzoic, protocatechuic, gallic, vanillic and syringic acid), hydroxybenzaldehydes (4-hydroxybenzaldehyde and 3,4-dihydroxybenzaldehyde) and cinnamic acid derivatives (o-coumaric, p-coumaric, caffeic, ferulic, sinapic and chlorogenic acid) were extracted. Cyclic addition of binary extraction solvent system based on methanol:water (1:1, v/v) and methanol/ammonia aqueous solution was used for extraction at 40MPa and 80 degrees C. The p-hydroxybenzoic, protocatechuic, vanillic, syringic, caffeic and chlorogenic acid; 4-hydroxybenzaldehyde and 3,4-dihydroxybenzaldehyde were identified by HPLC-electrospray mass spectrometry in SPE/SFE extracts of acid hydrolyzates of microalga (Spongiochloris spongiosa) and cyanobacterial strains (Spirulina platensis, Anabaena doliolum, Nostoc sp., and Cylindrospermum sp.). For the identification and quantification of the compounds the quasi-molecular ions [M-H](-) and specific fragments were analysed by quadrupole mass spectrometry analyzer. Our analysis showed that the microalgae and cyanobacteria usually contained phenolic acids or aldehydes at microg levels per gram of lyophilized sample. The proposed SPE/SFE extraction method would be useful for the analysis of different plant species containing trace amount of polar fraction of phenols.

Ultrahigh-pressure liquid chromatography of isoflavones and phenolic acids on different stationary phases.

Complete separation of aglycones and glucosides of selected isoflavones (genistin, genistein, daidzin, daidzein, glycitin, glycitein, ononin, sissotrin, formononetin, and biochanin A) was possible in 1.5 min using an ultrahigh-pressure liquid chromatography (U-HPLC) on a different particular chemically modified stationary phases with a particle size under 2 microm. In addition, selected separation conditions for simultaneous determination of isoflavones together with a group of phenolic acids (gallic, protocatechuic, p-hydroxybenzoic, vanillic, caffeic, syringic, p-coumaric, ferulic, and sinapic acid) allowed separation of all 19 compounds in 1.9 min. Separations were conducted on a non-polar reversed phase (C(18)) and also on more polar phases with cyanopropyl or phenyl groups using a gradient elution with a mobile phase consisting of 0.3% aqueous acetic acid and methanol. Chromatographic peaks were characterised using parameters such as resolution, symmetry, selectivity, etc. Individual substances were identified and quantified using UV-vis diode array detector at wavelength 270 nm. Limits of detection (3S/N) were in the range 200-400 pg ml(-1). Proposed U-HPLC technique was used for separation of isoflavones and phenolic acids in samples of plant materials (Trifolium pratense, Glycine max, Pisum sativum and Ononis spinosa) after acid hydrolysis of the samples and modified Soxhlet extraction.

[Modern instrumentation for the studies of isoflavones]. / Moderní instrumentální metody studia isoflavonu.

Isoflavones belong to the natural substances exhibiting a number of physiological effects in living organisms. The substances are synthesized in plant tissues as protective agents against biotic stress (i.e. bacterial infection). Isoflavones are also an important dietary constituent in human nutrition. The review discusses modern trends in the studies of isoflavones in plant materials and foodstuffs and procedures for chemical analyses of isoflavones in human body fluids and plant tissues. Highly effective extraction and purification techniques, i.e. solid-phase extraction (SFE), accelerated-solvent extraction (ASE), and Soxhlet extraction, are presented. Latest procedures in chromatographic separation of isoflavones that apply different types of stationary phases are described. Immunochemical analysis, electrochemical sensing of isoflavones, spectrometric and other analytical techniques and their applications are also mentioned. Special attention is focused on a highly selective and sensitive technique of mass spectrometry and its application for identification of isoflavones and their glucosides in plants. Studies of interactions of isoflavones with cell receptors and a number of biologically active substances such as DNA and proteins are described. The reason for the presentation of the review was not to give a full overview of the presented topics but mainly to show modern and the most recent methods in the studies of isoflavones.

Nontoxic combretafuranone analogues with high in vitro antibacterial activity.

A library of thirty two 3,4-diphenylfuranones related to both combretastatin A-4 and antifungal 5-(acyloxymethyl)-3-(halophenyl)-2,5-dihydrofuran-2-ones was prepared. Cytotoxic effects on a panel of cancer and normal cell lines and antiinfective activity were evaluated, and the data were complemented with tests for the activation of caspase 3 and 7. High cytotoxicity was observed in some of the halogenated analogues, eg. 3-(3,4-dichlorophenyl)-4-(4-methylphenyl)-2,5-dihydrofuran-2-one with IC50 0.12-0.23 µM, but the compounds were also highly toxic against non-malignant control cells. More importantly, notable antibacterial activity indicating G+ selectivity has been found in the 3,4-diarylfuranone class of compounds for the first time. Hydroxymethylation of furanone C5 knocked out cytotoxic effects (up to 40 µM) while maintaining significant activity against Staphylococcus strains in some derivatives. MIC95 of the most promising compound, 3-(4-bromophenyl)-5,5-bis(hydroxymethyl)-4-(4-methylphenyl)-2,5-dihydrofuran-2-one against S. aureus strain ATCC 6538 was 0.98 µM (0.38 µg/mL) and 3.9 µM (1.52 µg/mL) after 24 and 48 h, respectively.

Rapid-resolution HPLC with spectrometric detection for the determination and identification of isoflavones in soy preparations and plant extracts.

A rapid-resolution HPLC/UV-VIS DAD separation method (which takes <1 min) for the determination and identification of genistin, genistein, daidzein, daidzin, glycitin, glycitein, ononin, formononetin, sissotrin and biochanin A in fmol quantities in submicroliter sample volumes was optimized. A linear gradient elution (0 min 22% B, 1.0 min 80% B, 1.4 min 100% B, 1.8 min 22% B) using a mobile phase containing 0.2 % (v/v) acetic acid (solvent A) and methanol (solvent B) was applied on a Zorbax SB C18 column (1.8 microm particle size) at 80 degrees C. The method was verified using samples of bits of soy and methanolic extracts from Trifolium pratense, Iresine herbstii and Ononis spinosa plants. Pseudobaptigenin glucoside, irilone, prunetin, texasin, tlatlancuayin and other isoflavones, in addition to aglycones of isoflavones and their beta-glucosides and malonyl and acetyl derivatives, were identified by UV-VIS DAD and electrospray mass spectrometric (ESI-MS) detection in the extracts.

Hypericin and hyperforin production in St. John's wort in vitro culture: influence of saccharose, polyethylene glycol, methyl jasmonate, and Agrobacterium tumefaciens.

Influence of saccharose in the presence or absence of polyethylene glycol (PEG), methyl jasmonate, and an inactivated bacterial culture of Agrobacterium tumefaciens in cultivation medium on morphology of Hypericum perforatum L. and production of hypericin and hyperforin was studied under in vitro conditions. Production of hypericin and hyperforin was influenced by the presence of different concentrations of saccharose (10-30 g L(-1)) in cultivation medium. Addition of PEG (1.25-5 g L(-1)) in the presence of saccharose (10-30 g L(-1)) increased production of hypericin and hyperforin in the H. perforatum in vitro culture. Synthesis of hypericin and hyperforin was unchanged or reduced for most of the experimental plants at higher contents of PEG (10 and 15 g L(-1)). Concentrations of hypericin and hyperforin in the H. perforatum were on the order 100 and 103 microg g(-1) of dry plant material, respectively. Production of hypericin and hyperforin was stimulated either in the presence of a chemical elicitor (methyl jasmonate) or an inactivated bacterial culture of A. tumefaciens. Morphological changes induced by the abovementioned substances were observed and described in detail. The obtained results will be applied in experimental botany and in the technology of H. perforatum cultivation for pharmaceutical applications.

[Hypericin and hyperforin: bioactive components of St. John's Wort (Hypericum perforatum). Their isolation, analysis and study of physiological effect]. / Hypericin a hyperforin: biologicky aktivní komponenty trezalky teckované (Hypericum perforatum). Jejich izolace, analýza a studium fyziologických úcinku.

St. John's Wort (Hypericum perforatum L.) is commonly accepted as a medicinal plant. The data on the physiological activities of the individual substances that are produced in different organs of H. perforatum are well known at present. The highest attention is focused on the characterization and phytochemical properties of hypericin and hyperforin. These organic compounds are used as antidepressant, anticarcinogenic (photodynamic), antimicrobial and virostatic agents. The review paper surveys the present knowledge of chemical and analytical methods for their identification and quantification, physiological activity, and pharmacological and biomedical applications of hypericin and hyperforin.

Coronary angioplasty versus bypass surgery in patients > 70 years old matched for ventricular function.

OBJECTIVES: This study compared the relative risks and benefits of coronary angioplasty and coronary artery bypass graft surgery in patients > 70 years old. BACKGROUND: Few objective, comparative data exist to guide the clinician in the decision to use bypass surgery or angioplasty in elderly patients. METHODS: The study was a case-control, retrospective analysis of 195 consecutive patients who underwent bypass surgery in 1987 and 1988 and were compared with a concurrent cohort of 195 coronary angioplasty-treated patients. The groups were matched for left ventricular function, age and gender mix. RESULTS: The in-hospital morbidity and mortality rates were significantly lower in the coronary angioplasty-treated patients. Mean postprocedural hospital stay was 4.8 and 14.3 days for angioplasty and surgical group patients, respectively (p < 0.001). In-hospital death occurred in 2% of angioplasty-treated patients compared with 9% of surgically treated patients (p = 0.007). Serious in-hospital stroke occurred in no patient in the angioplasty group and in 5% of patients in the surgical group (p < 0.0001). Q wave infarction occurred in 1% of angioplasty-treated patients and 6% of bypass-treated patients (p = 0.01). The 5-year actuarial survival rate was similar in the two groups: 63% in the angioplasty group, 65% in the bypass group (p = NS). However, surgical group patients experienced less recurrent angina, required fewer repeat revascularization procedures and had fewer Q wave infarctions during follow-up compared with angioplasty group patients. CONCLUSIONS: When performed in patients > 70 years old, angioplasty and coronary bypass surgery result in similar long-term survival rates but otherwise distinctly different clinical courses.

The initial electrocardiogram during admission for myocardial infarction. Use as a predictor of clinical course and facility utilization.

The first electrocardiogram obtained on presentation for suspected myocardial infarction was examined for its usefulness in predicting clinical course and facility use. We studied 221 patients consecutively admitted to a nonuniversity hospital coronary care unit. High-risk patients were identified if the electrocardiographic diagnoses included myocardial infarction, ischemia, left ventricular hypertrophy, left bundle-branch block, or paced rhythm. These 63 patients (29% of total) had significantly greater incidences of serious events, need for procedures, and death than low-risk patients whose initial electrocardiograms did not carry the above diagnoses. Patients with a low-risk initial electrocardiogram may not require the facilities of a coronary care unit and perhaps could be safely observed in an intermediate care area. However, many hospitals do not have an intermediate care facility available, and in those that do, daily costs may not be markedly different than for treatment in a coronary care unit. Whether these low-risk patients could be safely treated in general medicine beds, where potential cost savings would be much greater, is unknown.

A randomized, controlled trial of the effects of remote, intercessory prayer on outcomes in patients admitted to the coronary care unit.

CONTEXT: Intercessory prayer (praying for others) has been a common response to sickness for millennia, but it has received little scientific attention. The positive findings of a previous controlled trial of intercessory prayer have yet to be replicated. OBJECTIVE: To determine whether remote, intercessory prayer for hospitalized, cardiac patients will reduce overall adverse events and length of stay. DESIGN: Randomized, controlled, double-blind, prospective, parallel-group trial. SETTING: Private, university-associated hospital. PATIENTS: Nine hundred ninety consecutive patients who were newly admitted to the coronary care unit (CCU). INTERVENTION: At the time of admission, patients were randomized to receive remote, intercessory prayer (prayer group) or not (usual care group). The first names of patients in the prayer group were given to a team of outside intercessors who prayed for them daily for 4 weeks. Patients were unaware that they were being prayed for, and the intercessors did not know and never met the patients. MAIN OUTCOME MEASURES: The medical course from CCU admission to hospital discharge was summarized in a CCU course score derived from blinded, retrospective chart review. RESULTS: Compared with the usual care group (n = 524), the prayer group (n = 466) had lower mean +/- SEM weighted (6.35 +/- 0.26 vs 7.13 +/- 0.27; P=.04) and unweighted (2.7 +/- 0.1 vs 3.0 +/- 0.1; P=.04) CCU course scores. Lengths of CCU and hospital stays were not different. CONCLUSIONS: Remote, intercessory prayer was associated with lower CCU course scores. This result suggests that prayer may be an effective adjunct to standard medical care.

Homocysteine Induced Cerebrovascular Dysfunction: A Link to Alzheimer's Disease Etiology.

A high serum level of homocysteine, known as hyperhomocystenemia (HHcy) is associated with vascular dysfunction such as altered angiogenesis and increased membrane permeability. Epidemiological studies have found associations between HHcy and Alzheimer's disease (AD) progression that eventually leads to vascular dementia (VaD). VaD is the second most common cause of dementia in people older than 65, the first being AD. VaD affects the quality of life for those suffering by drastically decreasing their cognitive function. VaD, a cerebrovascular disease, generally occurs due to cerebral ischemic events from either decreased perfusion or hemorrhagic lesions. HHcy is associated with the hallmarks of dementia such as tau phosphorylation, Aß aggregation, neurofibrillary tangle (NFT) formation, neuroinflammation, and neurodegeneration. Previous reports also suggest HHcy may promote AD like pathology by more than one mechanism, including cerebral microangiopathy, endothelial dysfunction, oxidative stress, neurotoxicity and apoptosis. Despite the corelations presented above, the question still exists - does homocysteine have a causal connection to AD? In this review, we highlight the role of HHcy in relation to AD by discussing its neurovascular effects and amelioration with dietary supplements. Moreover, we consider the studies using animal models to unravel the connection of Hcy to AD.

Mexiletine effects on theophylline disposition.

The effect of mexiletine administration on steady-state plasma theophylline concentrations was studied in eight normal healthy men in a prospective open label nonrandomized two-way crossover trial. Repeated doses of 300 mg of sustained-release theophylline were given every 12 hours for 9 days. Mexiletine hydrochloride, 200 mg every 8 hours, was given for five consecutive doses starting on the morning of day 6. Mexiletine increased theophylline levels in all subjects. Mean predose (trough) levels rose from 8.1 +/- 0.1 microgram.ml-1 to 13.4 +/- 0.6 micrograms.ml-1 and AUC(0-12) from 96.8 +/- 9.1 to 160.2 +/- 3.7 micrograms.ml-1.hr. Plasma clearance was reduced by mexiletine from 44.7 +/- 5.1 to 25.4 +/- 1.2 ml.hr-1. Both N-demethylated metabolites of theophylline were decreased by 60% by mexiletine, whose levels remained within its therapeutic range. Theophylline levels returned to pre-mexiletine values when this drug was discontinued. Mexiletine reduces theophylline clearance and increases its plasma concentration by inhibiting N-demethylation of theophylline. Plasma theophylline levels should be monitored when mexiletine is added.

Sarcoidosis mimicking cor triatriatum. Echolucency of adenopathy due to sarcoidosis.

A 20-year-old man with sarcoidosis presented with the echocardiographic findings of cor triatriatum. Computed tomography of the chest and digital subtraction angiography of the heart revealed that the patient had massive retrocardiac lymphadenopathy and normal cardiac anatomy. This is the first report demonstrating the echolucency of sarcoid lymphadenopathy and the mimicking of cor triatriatum by such adenopathy fortuitously positioned.

Prolonged and functional survival with the asplenia syndrome.

Asplenia syndrome is a rare congenital complex of splenic agenesis, cardiac malformation, and malposition of the abdominal viscera. Prolonged functional survival is very uncommon, with death usually caused by severe infection or congestive heart failure. A 21-year-old man with typical asplenia syndrome who is currently asymptomatic and in his third year of college is described.

Lack of reproducibility of frequency versus time domain signal-averaged electrocardiographic analyses and effects of lead polarity in coronary artery disease.

Time domain signal-averaged electrocardiographic parameters have been found to be consistent over the short term. Similar data have not been verified for the frequency domain. In addition, the effects of lead polarity changes on time domain or frequency domain parameters are not known. Signal-averaged electrocardiography was performed on 15 patients using the Arrhythmia Research Technology model 1200 EPX with orthogonal X, Y and Z leads. Time domain variables included filtered QRS, root-mean-square voltage of the last 40 ms of the QRS and duration of signals less than 40 microV. Two frequency domain area ratios were analyzed: a 140-ms window including last 40 ms of QRS + 100 ms of ST; and a 140-ms window beginning at QRS onset (both 20 to 50/0 to 20 Hz). Values were compared for each lead, as well as for a vector composite and the arithmetic mean of the 3 leads. Each patient underwent 3 studies performed 5 minutes apart. The polarity of the X lead was reversed between the first and second studies, and then returned to standard for the third study. Time domain variables correlated closely among the studies irrespective of lead polarity, with r values of 0.993 to 1.000 (p less than 0.0001). Frequency domain parameter correlation was much poorer, with r values as low as 0.276. Frequency domain correlations between tests 1 and 2 and 2 and 3 (mean r = 0.778) were poorer than between tests 1 and 3 (mean r = 0.829), implying an effect of lead polarity change.(ABSTRACT TRUNCATED AT 250 WORDS)

One-year outcomes of diabetic versus nondiabetic patients with non-Q-wave acute myocardial infarction treated with percutaneous transluminal coronary angioplasty.

Risk factors and outcomes associated with non-Q-wave myocardial infarction (MI) in diabetics and nondiabetics were analyzed for 376 consecutive patients, 77 with diabetes (20%) and 299 nondiabetics (80%), who had non-Q-wave MI and had percutaneous transluminal coronary angioplasty (PTCA) performed before discharge from hospital during the period from January 1992 to February 1996. Diabetics were slightly older (64 +/- 10 years vs 61 +/- 12 years, p <0.053), had more prior coronary artery bypass grafting (CABG) surgery (27% vs 12%, p <0.001), and hypertension (77% vs 49%, p <0.001). There was no significant difference in unstable angina, saphenous vein graft PTCA, single versus multiple vessel disease, or history of MI. PTCA success rates for diabetics versus nondiabetics were similar (96% vs 97%, p = NS). In-hospital complications such CABG, recurrent MI, repeat PTCA, stroke, and death were not statistically significant between the 2 groups. At 1-year follow-up, survival in diabetics (92%) was similar to nondiabetics (94%, p = NS), although event-free survival (PTCA, CABG, MI, death) was worse in diabetics (55% vs 67% for nondiabetics, p <0.05). Although diabetic patients with non-Q-wave MI represent a cohort with more risk factors for poor outcome, aggressive in-hospital revascularization with PTCA results in an excellent short-term outcome as well as 1-year survival similar to the nondiabetic patients. However, total events at 1-year follow-up are more common in the diabetic patients, suggesting that more aggressive screening and therapy in follow-up may be warranted, and that a diabetic with non-Q-wave MI will require increased utilization of cardiovascular resources in the first year after the event.

Gender-related differences in reperfusion treatment allocation and outcome for acute myocardial infarction.

Gender-related differences in outcome after myocardial infarction may relate to biased treatment allocation. To address this concern we analyzed 573 patients presenting with ST-segment elevation acute myocardial infarction (AMI), and treated within 6 hours with reperfusion therapy. Two-hundred eighty patients (49%) received direct coronary angioplasty, whereas 293 (51%) received thrombolytics followed by angioplasty (p = NS). Seventy-four percent were men and 26% were women (p = NS for differences in sex distribution between the 2 treatment groups). Women were older in both groups (p < 0.01). Inferior AMI was seen more often in women (64% of direct angioplasty, 71% of lytic first) than in men (51% and 59%, respectively; p < 0.03). There was no gender-related differences in presence of multivessel coronary artery disease, prior AMI, prior bypass surgery, baseline ejection fraction, percentage of patients with ejection fraction < or = 40%, number of narrowings dilated, or angioplasty success. Patients who underwent direct angioplasty had more multivessel disease (p < 0.001) and prior coronary artery bypass surgery (p = 0.002). After a mean follow-up of 129 +/- 113 weeks, no gender-related differences were seen in the need for cardiac catheterization, documented restenosis, AMI, coronary artery bypass surgery, clinical ischemia, or death. Patients treated with direct angioplasty were more likely to undergo coronary artery bypass surgery (p < 0.05) or to die (p < 0.01). Thus, women undergoing reperfusion therapy for ST-segment elevation were older than men, with a higher frequency of inferior wall AMI. No specific gender-related bias in treatment allocation was evident.