In vitro blood compatibility and in vitro cytotoxicity of amphiphilic poly-N-vinylpyrrolidone nanoparticles.

This study focused on defining the in vitro behavior of amphiphilic poly-N-vinylpyrrolidone (Amph-PVP) nanoparticles toward whole blood, blood plasma and blood cells in order to assess nanoparticle blood compatibility. In addition, possible effects on endothelium cell growth/viability were evaluated. The Amph-PVP nanoparticles were formed via self-assembling in aqueous media and composed of a hydrophobic alkyl core and a hydrophilic PVP outer shell. Their blood compatibility was evaluated by investigating their effect on red blood cells (RBCs) or erythrocytes, white blood cells (WBCs) or leukocytes, platelets (PLTs) and on complement system activation. Our results clearly demonstrate that the Amph-PVP nanoparticles are stable in presence of blood serum, have no significant effects on the function of RBCs, WBCs, PLTs and complement system activation. The Amph-PVP nanoparticles did not show considerable hemolytic or inflammatory effect, neither influence on platelet aggregation, coagulation process, or complement activation at the tested concentration range of 0.05-0.5 mg/ml. The Amph-PVP nanoparticles did not exhibit any significant effect on HMEC-1 microvascular skin endothelial cells' growth in in vitro experiments. The excellent blood compatibility of the Amph-PVP nanoparticles and the lack of effect on endothelium cell growth/viability represent a crucial feature dictating their further study as novel drug delivery systems.

Common mental disorders and association with telomere length.

Telomeres are repeated 5'-TTAGGG-3' sequences at the end of chromosomes, which maintain genomic stability. Their length is related to a number of diseases that affect humans. Apart from cancer, cardiovascular diseases, diabetes and other, telomere length has been associated with chronic diseases. Chronic mental illness includes various types of mental disorders with the most common being depression, schizophrenia and stress-anxiety. The aim of this review is to summarize the current state of knowledge on the role of telomeres in these disorders and to compare telomere length variations in patients receiving medication and patients not taking treatment. Most studies report reduced telomere length in patients suffering from mental disorders, compared to the general population. Since the factors that can affect telomere length are various, more experiments and investigations are required to understand the general impact of different factors on telomere length.

Simulating real-life exposures to uncover possible risks to human health: A proposed consensus for a novel methodological approach.

In real life, consumers are exposed to complex mixtures of chemicals via food, water and commercial products consumption. Since risk assessment usually focuses on individual compounds, the current regulatory approach doesn't assess the overall risk of chemicals present in a mixture. This study will evaluate the cumulative toxicity of mixtures of different classes of pesticides and mixtures of different classes of pesticides together with food additives (FAs) and common consumer product chemicals using realistic doses after long-term exposure. Groups of Sprague Dawley (CD-SD) rats (20 males and 20 females) will be treated with mixtures of pesticides or mixtures of pesticides together with FAs and common consumer product chemicals in 0.0, 0.25 × acceptable daily intake (ADI)/tolerable daily intake (TDI), ADI/TDI and 5 × ADI/TDI doses for 104 weeks. All animals will be examined every day for signs of morbidity and mortality. Clinical chemistry hematological parameters, serum hormone levels, biomarkers of oxidative stress, cardiotoxicity, genotoxicity, urinalysis and echocardiographic tests will be assessed periodically at 6 month intervals. At 3-month intervals, ophthalmological examination, test for sensory reactivity to different types of stimuli, together with assessment of learning abilities and memory performance of the adult and ageing animals will be conducted. After 24 months, animals will be necropsied, and internal organs will be histopathologically examined. If the hypothesis of an increased risk or a new hazard not currently identified from cumulative exposure to multiple chemicals was observed, this will provide further information to public authorities and research communities supporting the need of replacing current single-compound risk assessment by a more robust cumulative risk assessment paradigm.

Feasibility of hair sampling to assess levels of organophosphate metabolites in rural areas of Sri Lanka.

Measuring chronic pesticide exposure is important in order to investigate the associated health effects. Traditional biological samples (blood/urine) are difficult to collect, store and transport in large epidemiological studies in settings such as rural Asia. We assessed the acceptability of collecting hair samples from a rural Sri Lankan population and found that this method of data collection was feasible. We also assessed the level of non-specific metabolites (DAPS) of organophosphate pesticides in the hair samples. The median concentration (pg/mg) of each DAP was: diethyl phosphate: 83.3 (IQI 56.0, 209.4); diethyl thiophosphate: 34.7 (IQI 13.8, 147.9); diethyl dithiophosphate: 34.5 (IQI 23.4, 55.2); and dimethyl phosphate: 3 (IQI 3, 109.7). Total diethylphosphates were recovered in >80% of samples and were positively correlated with self-reported pesticide exposure.

Dialkyl phosphates in amniotic fluid as a biomarker of fetal exposure to organophosphates in Crete, Greece; association with fetal growth.

The aim of this study was to evaluate fetal exposure to organophosphate pesticides (OPs) by measuring their non-specific dialkyl-phosphate metabolites (DAPs) in amniotic fluid (AF), and to examine the potential association between prenatal exposure and fetal growth. AF samples were collected from 415 women during the second gestational trimester. The determined OPs metabolites were DMP, DMTP, DEP, DETP, and DEDTP. DAPs were extracted by liquid-solid extraction, derivatized and analyzed by gas chromatography-mass spectrometry. 97.8% of AF samples were positive for at least one DAP. DAPs levels did not differ between urban and rural areas. Macrosomic neonates have significantly higher sum levels of DMPs (p=0.043), which exerted a linear positive association with birth-weight centile (b=4.43, p=0.016). Conclusively, as DAPs are detectable in AF they may be used as a potential biomarker of fetal exposure to OPs. Sum levels of DMPs appear to be associated with birth weight independently of other covariates.

Comparative evaluation of xenobiotics in human and dietary milk: persistent organic pollutants and mycotoxins.

Publications produced over the past 20 years regarding the concentration of xenobiotics in human and dietary milk were evaluated, focusing primarily on persistent organic pollutants (e.g. polychlorinated biphenyls, flame retardants), pesticides (e.g organochlorine) and mycotoxins. In general, countries of low industrialization rate present low levels of dietary milk contamination with dioxins compared to those with high rate of industrialization. According to published data, the most common persistent organic pollutants detected in breast and dietary milk are dichlorodiphenyltrichloroethane compounds, hexachlorocyclohexane, and hexachlorobenzene. Even though the potential risks of persistent organic pollutants in human milk have been acknowledged, the beneficial effect of breastfeeding as the optimal food source for newborn babies should not be disregarded. Especially when sharing information with the general public, it should be made clear that the presence of dioxins and persistent organic pollutants in human milk is not an indication for avoiding breastfeeding. The implications of xenobiotics in human and dietary milk is a matter of growing importance and warrants future work given its important health effects.

Hair nicotine/cotinine concentrations as a method of monitoring exposure to tobacco smoke among infants and adults.

In this pilot study, we examined the validity and usefulness of hair nicotine-cotinine evaluation as a biomarker of monitoring exposure to tobacco. Head hair samples were collected from 22 infants (<2 years of age) and 44 adults with different exposures to tobacco (through either active or passive smoking) and analyzed by liquid chromatography-mass spectrometry (LC-MS) for nicotine and cotinine. Hair samples were divided into three groups, infants, passive smoker adults and active smoker adults, and into eight subgroups according to the degree of exposure. The limit of quantification (LOQ) was 0.1 ng/mg for nicotine and 0.05 ng/mg for cotinine. Mean recovery was 69.15% for nicotine and 72.08% for cotinine. The within- and between-day precision for cotinine and nicotine was calculated at different concentrations. Moreover, hair nicotine and cotinine concentrations were highly correlated among adult active smokers (R (2) = 0.710, p < 0.001), among adult nonsmokers exposed to secondhand smoke (SHS; R (2) = 0.729, p < 0.001) and among infants (R (2) = 0.538, p = 0.01). Among the infants exposed to SHS from both parents the noted correlations were even stronger (R (2) = 0.835, p = 0.02). The above results identify the use of hair samples as an effective method for assessing exposure to tobacco, with a high association between nicotine and cotinine especially among infants heavily exposed to SHS.

Is hair analysis for dialkyl phosphate metabolites a suitable biomarker for assessing past acute exposure to organophosphate pesticides?

In the present paper, the possibility to use dialkyl phosphate metabolites (DAPs) hair segmental analysis as a biomarker of past acute exposure to organophosphates is examined. Hair samples of four acute poisoning survivors were collected and segmental hair analysis was performed. The total hair samples were divided to 1 cm segments and analysed by gas chromatography-mass spectrometry (GC-MS) for the presence of four DAP metabolites, dimethyl phosphate (DMP), diethyl phosphate (DEP), diethyl thiophosphate (DETP) and diethyl dithiophosphate (DEDTP). Results were examined under the light of pesticide type and time of hair sample collection. Although DAPs were detected all along the hair shaft, higher concentrations (peaks) were detected in the segments proximate to the suicide period. It was also observed that the elevated concentrations of the present metabolites corresponded to the ones produced by the ingested parent compound. Conclusively, measurements of DAPs in the appropriate hair segments of OP-poisoned patients can be used for assessing past acute exposure to organophosphates in certain cases.

Determination of dialkyl phosphates in human hair for the biomonitoring of exposure to organophosphate pesticides.

A new, simple, fast and sensitive method that enables the measurement of four dialkyl phosphates (DAPs) in human head hair is presented in the current study. The dialkyl phosphates, dimethyl phosphate (DMP), diethyl phosphate (DEP), diethyl thiophosphate (DETP) and diethyl dithiophosphate (DEDTP) are non-selective metabolites of the organophosphate pesticides (OPs). The extraction of DAPs from hair matrix was achieved by one step methanolic extraction. Head hair samples from general population and population occupationally exposed to OPs were analysed using gas chromatography-mass spectrometry (GC-MS) after derivatization with pentafluorobenzylbromide. The recovery of the target compounds was estimated at 84.3% for DMP, 116.1% for DEP, 109.0% for DETP and 91.5% for DEDTP. The limit of quantitation (LOQ) and detection (LOD) was 20 and 6 pg/mg for DMP, 10 and 5 pg/mg for DEP and DETP and 5 and 3 pg/mg for DEDTP, respectively. With-run and between-run precision as well as accuracy was estimated. The percentage of positive hair samples for DMP, DEP, DETP and DEDTP for the group of general population was 63.0%, 96.3%, 66.7%, and 70.4% respectively. The samples from the group with occupational exposure were positive for all dialkyl phosphates analysed. The median concentrations for DMP were 165.0 and 181.7 pg/mg, for DEP were 51.2 and 812.9 pg/mg, for DETP were 54.0 and 660.1 pg/mg, and for DEDTP were 40.0 and 60.6 pg/mg for the general population group and the group with occupational exposure respectively. Significant differences in the levels of the total dialkyl phosphates amongst exposed and not exposed groups were observed (p<0.001). More specifically, the total ethyl phosphate (DEPs) and DAPs median concentrations were 119.5 and 301.5 pg/mg for the general population group and 1498.8 and 1694.4 pg/mg for the group with occupational exposure.

Biomarker evaluation of Greek adolescents' exposure to secondhand smoke.

Exposure to secondhand smoke (SHS) is a significant threat to public health, and represents a danger for both the development and health status of children and adolescents. Taking the above into account, our aim was to quantify Greek adolescents' exposure to SHS using serum cotinine levels. During 2006, 341 adolescents aged 13-17 were randomly selected from high schools in Heraklion and agreed to participate as part of the European Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) study. Blood samples were drawn from a random sample of 106 adolescents, while serum cotinine/nicotine concentrations were measured by Gas Chromatography-Mass Spectrometry (GC-MS). The mean levels of serum cotinine and nicotine were calculated at 1.60 +/- 2.18 ng/mL and 4.48 +/- 4.00 ng/mL, respectively, while 97.7% of the non-smoker adolescents were found to have measureable levels of serum cotinine indicating exposure to SHS. The analysis revealed that their paternal (p = .001) and maternal smoking habits (p = .018) as also the existence of a younger brother or sister (p = .008) were the main modifiers of SHS exposure during adolescence. Conclusively, almost all of the measured Greek adolescents were exposed to SHS, even when their parents were non-smokers. This finding indicates the need for both community and school-based educational programmes as also the implementation of a comprehensive ban on smoking in public places.

Dialkyl phosphates in meconium as a biomarker of prenatal exposure to organophosphate pesticides: a study on pregnant women of rural areas in Crete, Greece.

The authors developed a sensitive analytical method for the determination of dialkyl phosphates (DAPs) in meconium. This method was applied to determine the DAPs, which are non-specific metabolites of the organophosphate pesticides (OPs), in meconium of newborns by mothers who live in rural areas in Crete, Greece. DAPs are considered as biomarkers of exposure to OPs. Meconium is produced in the foetus at approximately 16 weeks of gestation and it acts as a repository of many xenobiotics. The determined organophosphate metabolites were dimethylphosphate (DMP), diethylphosphate (DEP), dimethylthiophosphate (DMTP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP). The DAPs were extracted from meconium by liquid-solid extraction, derivatized, and analysed by gas chromatography-mass spectrometry (GC-MS). The mean percentile recoveries were 76.9%, 65.2%, 94.1%, 109.4%, and 107.2% for DMP, DEP, DMTP, DETP, and DEDTP, respectively. The percentage of positive samples was 92.1% for DMP, 36.8% for DEP, 60.5% for DMTP, 63.2% for DETP, and 57.9% for DEDTP. Mean (+/- standard deviation) and the range concentrations of the positive samples (ng g(-1)) were 126.74 +/- 142.73 (10.64-739.45), 11.46 +/- 20.43 (1.50-79.14), 215.05 +/- 187.34 (8.54-662.16), 4.92 +/- 5.09 (1.25-19.04), and 1.84 +/- 2.07 (0.5-8.04) for DMP, DEP, DMTP, DETP, and DEDTP, respectively. Statistical analysis revealed no significant difference in meconium levels between high- and low-risk groups of exposure of pregnant women. However, the results of this study demonstrate that DAPs in meconium may be considered as a potential biomarker for the assessment of foetal exposure to organophosphate pesticides.

Relation of PON1 and CYP1A1 genetic polymorphisms to clinical findings in a cross-sectional study of a Greek rural population professionally exposed to pesticides.

Allelic variants of CYP1A1 and PON1 have been extensively studied as susceptibility factors in toxic response, although little is known about the role of these variants as risk factors for the plethora of diseases appearing in the human population. In this study we investigated the hypothesis of correlation of CYP1A1 and PON1 enzymes with the incidence of various medical examination findings in a Greek rural population professionally exposed to a variety of pesticides. The medical history of 492 individuals, randomly selected for the total population of 42,000, was acquired by interviews and their genotype determined for the CYP1A1*2A, PON1 M/L and PON1 Q/R polymorphisms. The assessment of exposure to pesticides of the population was verified by analytical methods. Analysis of the genetic data revealed that the allele frequencies of PON1 R, M and CYP1A1*2A alleles were 0.243, 0.39 and 0.107 respectively. The CYP1A1*2A polymorphism was found to have significant association with chronic obstructive pneumonopathy (p=0.045), peripheral circulatory problems (trend p=0.042), arteritis (p=0.022), allergies (trend p=0.046), hemorrhoids (trend p=0.026), allergic dermatitis (p=0.0016) and miscarriages (p=0.012). The PON1 Q/R polymorphism was found to have significant association with hypertension (p=0.046) and chronic constipation (p=0.028) whereas, the L/M polymorphism, with diabetes (p=0.036), arteritis (trend p=0.022) and hemorrhoids (trend p=0.027). Our results demonstrated an association between the CYP1A1/PON1 polymorphisms and several medical examination findings, thus indicating the possible involvement of the human detoxification system to health effects in a rural population exposed professionally to pesticides.

Assessment of levels of organochlorine pesticides and their metabolites in the hair of a Greek rural human population.

We present the assessment of chronic exposure of the rural population of Helia Peloponnesus, Greece to banned organochlorine pesticides, hexachlorocyclohexane (HCH), and 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT), using hair analysis. A total of 222 head hair samples were collected and analyzed for the presence of those organochlorine pesticides and their metabolites or isomers. Gas chromatography coupled to mass spectrometry was used to measure the levels of the pollutants. The median concentrations of alpha-HCH, hexachlorobenzene, lindane, ortho para 1,1-dichloro-2,2-bis-(4-chlorophenyl)ethylene (opDDE), para para 1,1-dichloro-2,2-bis(4-chlorophenyl)ethylene (ppDDE), ortho para 1,1-dichloro-2,2-bis(4-chlorophenyl)ethane (opDDD), para para 1,1-dichloro-2,2-bis(4-chlorophenyl)ethane (ppDDD) + ortho para 1,1,1-trichloro-2,2-bis(4-chlorophenyl)-ethane, and para para 1,1,1-trichloro-2,2-bis(4-chlorophenyl)-ethane were determined at 40.4, 19.7, 124.2, 6.2, 7.8, 73.1, 8.0, and 5.7 pg/mg. The median concentration of total HCHs and DDTs were 117.8 pg/mg and 9.4 pg/mg, respectively. The levels of total HCHs were much higher than the levels of DDTs in the hair samples of the studied population. This may be attributed to the presence of lindane, a pesticide officially banned in 2002. It is interesting to see that DDTs are still traced in samples despite their use being banned for more than three decades. There was no difference in the levels of the detected pesticides in hair sampled from men or women. The concentration of HCHs remains high and relatively stable across the age groups, suggesting constant exposure until very recently. The concentration of the total DDTs and the parent compound, pp-DDT presents a statistically significant decreasing trend across the age groups.

Pesticide levels in head hair samples of Cretan population as an indicator of present and past exposure.

In the present work we assessed chronic exposure of different working population groups of Messara and Sitia districts, Crete, Greece, to common currently used pesticides (diazinon, fenthion, methyl parathion and malathion) and two banned pesticides hexachlorocyclohexane (HCH) and DDT. The study population (211 persons, 110 females and 101 males) was divided to three groups; people working in greenhouses, animal breeders and people working in open cultivations. Methanolic extraction of pulverized hair was used for organophosphate pesticides extraction, followed by liquid-liquid extraction with water-ethyl acetate as a clean up step. The extraction of organochlorine pollutants was performed by acidic hydrolysis of the hair matrix followed by liquid-liquid and solid phase extraction. The levels of the aforementioned pesticides were measured by GC-ECD and gas chromatography-mass spectrometry (GC-MS). The median concentrations of a-HCH, HCB, lindane, opDDE, ppDDE, opDDD, ppDDD + opDDT and ppDDT were determined at 7.2, 2.2, 70.2, 2.7, 5.7, 3.1, 2.6 and 23.2 pg/mg. The median concentration of total HCHs and DDTs detected in the three working groups were: 95.0 and 8.9 pg/mg for the greenhouse workers, 38.2 and 3.3 pg/mg for the animal breeders and 24.1 and 5.2 pg/mg for the open cultivation group. Ten head hair samples were positive for diazinon at a mean concentration of 2.8 pg/mg. Fenthion, methyl parathion and malathion were not detected. Our results demonstrated the ability to assess chronic human past pesticides exposure, offering valuable information to epidemiological clinical studies.

Legal issues of addiction assessment: the experience with hair testing in Greece.

The purpose of this paper is to present Greek law and legislation for crimes and felonies regarding drugs of abuse and the interpretation of hair testing results with respect to Greek law. Details (such as the process, the decision and the competence of the Court, the police record, the indictment, the expert reports, the defendant's individuality, the crimes and the penal confrontal and many others) from legal cases related to toxicomany and its judicial verification were collected and analysed. Laboratory data of cases concerning the laboratory evaluation of toxicomany in addicts and also occasionally the legal course of cases with addict defendants are presented. In four representative cases segmental hair analysis proved that, for as long as the individuals were imprisoned, findings with drug substances corresponding to that period were lesser or practically absent compared with samples corresponding to the time out of prison, which showed increased drug abuse. Hair analysis provides information on chronic exposure rather than acute poisoning. Its detection window varies from some days to months or even years. The procedure that the law lays down in many cases is insufficient and in most cases impossible to abide by. When the medical examiner is not able to decide if the claim of toxicomany is real, segmental hair analysis may be the only way to prove it. In other cases where the medical examiner is able to diagnose the addiction, a segmental hair analysis is necessary because it can show long-term drug abuse.

Penetration of newer quinolones in the empyema fluid.

The degree of penetration of newer quinolones into the pleural fluid has not been studied. The objective of the present study was to determine the degree to which moxifloxacin and levofloxacin penetrate into empyemic pleural fluid using a new rabbit model of empyema. An empyema was created via the intrapleural injection of turpentine (1 mL), followed 24 h later by instillation of 2 mL (1 x 10(10)) Escherichia coli bacteria (ATCC 35218) into the pleural space of New Zealand white rabbits. After an empyema was verified by thoracentesis and pleural fluid analysis, moxifloxacin and levofloxacin (25 mg.kg(-1) for both, i.v.) were administered. Antibiotic levels were determined in samples of pleural fluid and in blood collected serially over 12 h. Antibiotic levels were measured using HPLC. Each of the antibiotics penetrated well into the empyemic pleural fluid. Antibiotic penetration was the greatest for moxifloxacin (area under the curve (AUC) for pleural fluid/blood (AUCPF/AUCblood) ratio=1.37) followed by levofloxacin (ratio=1.13). The time to equilibration between the pleural fluid and blood antibiotic levels was more rapid for moxifloxacin (3.9 h) than for levofloxacin (4.4 h). With moxifloxacin, the peak pleural fluid concentration (Cmax,PF) was 2.77 microg.mL(-1) and occurred at a time to maximum pleural fluid concentration (Tmax,PF) of 6 h after infusion and decreased thereafter. The peak blood concentration (Cmax,blood) was 4.81 microg.mL(-1) at 1 h after administration. With levofloxacin, the peak pleural fluid level (Cmax,PF=1.39 microg.mL(-1)) occurred at 6 h (Tmax,PF=6 h) after infusion. The Cmax,blood was 1.88 microg.mL(-1) at 1 h after administration. In conclusion, differences were found in the degree of penetration of the two quinolones into infected pleural fluid in rabbits. The clinical significance of these differences is unknown. More studies are needed to evaluate the pharmacokinetic parameters in the pleural space in humans.

Three-year study of fenthion and dimethoate pesticides in olive oil from organic and conventional cultivation.

Residues of fenthion and dimethoate pesticides were determined in organic and conventional olive oils by liquid-liquid and solid-phase extractions with subsequent gas chromatography and mass spectrometric analysis. The olive oil samples were collected from Crete during 1997-99. The average concentrations of fenthion in conventional olive oils were 0.1222, 0.145 and 0.1702 mg x kg(-1), and for dimethoate were 0.0226, 0.0264 and 0.0271 mg x kg(-1) for 1997, 1998 and 1999, respectively. The average concentrations of fenthion in organic olive oils were 0.0215, 0.0099 and 0.0035 mg x kg(-1) for 1997, 1998 and 1999, while for dimethoate they were 0.0098, 0.0038 and 0.0010 mg x kg(-1), respectively. All the olive oils contained residue levels lower than the maximum residue levels according to the FAO/WHO Codex Alimentarius. The organic olive oil contained significantly lower concentrations of the two pesticides. The levels of fenthion and dimethoate in organic olive oils exhibited a decreasing trend following the implementation of the new cultivation method. We propose procedures that should be established in the organic cultivation in order to maximize its effectiveness.

Evaluation of the addiction history of a dead woman after exhumation and sectional hair testing.

In Greece, sectional hair analysis, in addition to clinical examination, has been used as a valuable tool for the confirmation of a person's history of drug use. The present report concerns the toxicologic analysis of the exhumed remains and hair samples of an 18-year-old woman. Postmortem toxicologic analysis of blood and urine confirmed recent opiate and cannabis use and indicated that death was associated with heroin abuse. Several months later, the woman's family asked for exhumation and reexamination of the body, insisting that the cause of death was homicide. The investigating judge ordered exhumation and new medicolegal examination of the body. The investigation of the drug profile along the hair shaft was undertaken by analyzing hair sections 1 cm from the hair root for morphine, 6-monoacetylmorphine, heroin, and cannabinoids. The total lengths of the hair samples ranged from 8 to 11 cm. The total morphine levels in the hair sections corresponding to the 3-month period before death were significantly lower (1.5-2.85 ng/mg) than those of the 4- to 10-month period before death (7.4-14.8 ng/mg). An interpretation of these results may be occasional drug use (with considerable attenuation of use during the last 3 months before death). Decrease of tolerance to heroin caused by abstinence and relapse in use could have been the cause of death.

Comparison of in vitro activities of amphotericin, clotrimazole, econazole, miconazole, and nystatin against Fusarium oxysporum.

The inhibitory activity of amphotericin B, clotrimazole, econazole, miconazole and nystatin was compared against Fusarium oxysporum f.sp. radicis-cucumerinum. The most efficient antifungal agent against the growth of Fusarium oxysporum was econazole, followed by clotrimazole, miconazole, amphotericin and nystatin. The ED50 and ED90 values were 0.053 and 1.002 ppm for econazole, 0.088 and 1.100 ppm for clotrimazole, 0.173 and 3.210 ppm for miconazole, 0.713 and greater than 48 ppm for amphotericin and 3.860 and 16.702 ppm for nystatin. The ED50 values of nystatin and amphotericin against spore germination of Fusarium oxysporum were determined at 3.1427 ppm and 8.3990 ppm respectively, nystatin was 2.76 times more effective than amphotericin, while no effect was observed after the addition of econazole, clotrimazole and miconazole. The tested azoles were more effective than amphotericin and nystatin on growth inhibition of Fusarium oxysporum but amphotericin and nystatin acted significantly better on spore germination of Fusarium.

The application of histochemical methods to the age evaluation of skin wounds: experimental study in rabbits.

Enzyme histochemical methods allow determination of wound age, especially in the range of a few hours, and are used to distinguish between postmortem and antemortem skin wounds. The methods are based on the determination of the presence and changes of the enzyme reaction in the wound area. Increased activity of nonspecific esterases was observed approximately 1 hour after wounding and was followed by an increase in adenosine triphosphatase at approximately 2 hours and alkaline phosphatase at approximately 3.5 hours. Maximum enzyme activity was reached for nonspecific esterases at 24 hours, for adenosine triphosphatase at 20 hours, and for alkaline phosphatase at 32 hours after wounding.