The β-adrenoceptors and catecholamine levels in lead poisoned rats

To investigate β-adrenoceptor dysfunction upon exposure to lead, we measured (a) β-adrenoceptor density in brain, heart, blood vessels and lymphocytes and (b) plasma catecholamine levels in rats with lead poisoning. Wistar rats were given drinking water containing lead acetate (2% w/v) for a period of 60 days. The radioligand [(125)I]iodocyanopindolol was used for determining the density of β-adrenoceptors in membrane fragments in vitro and a high performance liquid chromatography (HPLC) for measuring plasma catecholamine levels. Plasma norepinephrine levels were found to be significantly higher in lead-exposed rats than in control animals (4.69 ± 0.58 μg/l vs. 3.67 ± 0.53 μg/l, p < 0.01). In lead-exposed animals the density of β-adrenoceptors in brain (36%), heart (68%), blood vessels (57%) and lymphocytes (48%) was significantly less than in controls (p < 0.001), whereas the K(d) did not vary between the two groups. We have found that β-adrenoceptor dysfunction in lead-poisoned rats was brought about by a decline in β-adrenoceptor density.     

Environmental cadmium and lead exposure and anti-Müllerian hormone in pregnant women

BackgroundAnti-Müllerian Hormone (AMH) has been suggested as a marker for ovarian function. Cadmium and lead have been suggested to reduce female fecundity. In this study we aimed to investigate whether environmental exposure to cadmium and lead was associated with alterations in serum-AMH.Materials and methodThe associations between serum-AMH and whole blood cadmium or lead were investigated by general linear models in a population-based sample of 117 pregnant women.ResultsThe mean concentrations of blood cadmium and lead were 0.71 μg/L and 17.4 μg/L, respectively. The mean serum-AMH was 17.3 pmol/L. No association between lead and AMH was detected. In the cadmium analysis the adjusted mean AMH level (95% CI) in the highest exposure tertile was 12.4 (6.4;23.8) compared to 5.6 (2.7;11.4) in the lowest exposure tertile (p = 0.06).ConclusionThe study provides suggestive evidence that environmental exposure to cadmium, but not lead, may alter the level of AMH.     

Low levels of lead exposure induce oxidative damage and DNA damage in the testes of the frog Rana nigromaculata

We have investigated the chronic effects of low concentrations of lead (Pb) on oxidative damage and DNA damage in testes of the frog Rana nigromaculata. Sixty adult male frogs were randomly divided into six groups of ten. Based on the levels of the Integrated Wastewater Discharge Standard (GB 8978-1996) of China, five groups (II–VI) were treated by epidermal absorption with a PbNO₃ solution at concentrations of 0.1, 0.2, 0.4, 0.8, 1.6 mg/l, respectively. The first group (I), which served as a control, was treated with distilled water only. Thirty days after treatment, all frogs were sacrificed and the testis tissues removed for the measurement of malondialdehyde (MDA) and glutathione (GSH) levels. DNA damage, including indicators of damage rate, DNA tail length (TL), and DNA tail moment (TM), was also analyzed by comet assays. Our data suggest that MDA levels in all treatment groups and GSH levels in the 0.2–1.6 mg/l Pb groups increased significantly relative to the controls (P < 0.01). Treatment with Pb at concentrations >0.4 mg/l also increased DNA damage rate and TM, while TL increased when the Pb level was >0.2 mg/l (P < 0.01 for DNA damage rate and TM, P < 0.05 for TL). Positive correlations were also found between DNA damage levels in the testes and MDA levels (r = 0.796 for DNA damage rate, r = 0.811 for TL, r = 0.796 for TM; P < 0.01 for all) as well between MDA and GSH levels (r = 0.455, P < 0.05) in the testes. Results from MDA measurements indicated that Pb-induced DNA damage in the testes of R. nigromaculata was possibly due to oxidative damage. Taken together, we conclude that Pb can induce male reproductive toxicity in R. nigromaculata.     

Electroretinographic and Ophthalmologic Examinations in the Yucatan Micropig and in the Göttingen Minipig

Abstract

External gross observations of the eye and its adnexae, ocular reflexes, anterior ocular segment biomicroscopic examinations, fundic examinations performed with an indirect ophthalmoscope, and/or electroretinographic investigations (ERG) were carried out on 112 7-12-month-old Yucatan micropigs, on 18 6-8-week-old, and 81 2-10-month old Gottingen minipigs to evaluate the incidence of observed ocular abnormalities and to compare the ERG waves. A statistical comparison was performed for these findings.

The most important ocular defects were classified as remnants of embryological vascular tissue. The other findings were considered either as embryonic remnants or of nondeterminate etiology. The most noteworthy findings were, in decreasing order of incidence, for Yucatan micropigs, 6-8 week-old and 2-10-monfh-old Gottingen minipigs, respectively, hyaloid artery remnants (82.1%, 83.3%, and 46.3%), pupillary     

Flavonoids as lead compounds modulating the enzyme targets in Alzheimer’s disease

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder of the brain amongst the elderly. The various enzyme targets have been identified including cholinesterase (acetylcholinesterase and butyrylcholinesterase), beta secretase, glycogen synthase kinase-3β and NADPH that play an important role in the pathogenesis of AD. Flavonoids are the phytochemicals with wide range of potential therapeutic activities including AD. Naturally occurring flavonoids have been shown to produce the beneficial effects in experimental models of AD through multiple mechanisms. Accordingly, the naturally occurring flavonoids scaffolds have been modified or new flavonoids analogues have been synthesized to obtain effective drugs for AD management. The present review describes the enzyme targets-(of AD) modulating naturally/synthesized flavonoids which may be potentially useful in the management of AD.     

Synthesis and in vivo lead detoxification evaluation of Poly-α,β-dl-aspartyl-l-methionine

To increase the metal selectivity of polyaspartic acid, a so-called green chelant, poly-α,β-dl-aspartyl-l-methionine (PDM) was synthesized as a novel lead chelating agent. The phosphoric acid (80%) catalyzed thermal poly condensation of dl-aspartic acid provided poly succinimide, which was amidated with l-methionine to form PDM (MW: 29161). At the doses of 0.1, 1.0, and 10.0 nmol/kg, either by intraperitoneal injection (i.p.) or oral administration, PDM removed Pb from the spleens, hearts, and kidneys of mice, especially dose-dependently decreasing the accumulation of Pb in the brains, livers, and femurs of the mice, and did not interfere with the essential metals, including Cu, Fe, Mn, and Ca. Even at the dose of 0.1 nmol/kg, the i.p. injection of PDM removed Pb from the spleens, hearts, and kidneys of mice and increased the amount of urinary volume and urinary Pb, and the amount of fecal matter and the amount of fecal Pb, resulting in effective removal of Pb from the body of mice given Pb by i.p. injection. Our findings revealed that in aqueous solution PDM formed diverse nanospecies.     

Effect of lead exposure on some biological indices related to porphyrin metabolism and the activity of erythrocyte pyrimidine 5′-nucleotidase in the mice

The urinary excretion of -aminolevulinic acid (ALA) and coproporphyrin (CP) isomers (I and III) was investigated in mice exposed to lead in drinking water (200 and 500 ppm) for 14 or 30 days. Furthermore, the inhibition of pyrimidine 5-nucleotidase (P5N) and -aminolevulinic acid dehydratase (ALAD) was studied in the same mice. In the lead-exposed mice, urinary excretion of ALA was significantly elevated (12-times control), while that of CP remained unchanged. This discrepancy on the urinary excretion may be of interest. The P5N activity in both erythrocytes and bone marrow cells was significantly inhibited by exposure to lead. However, the degree of inhibition was greater in the erythrocytes (45% inhibition) than in the bone marrow cells (25% inhibition). The inhibition of ALAD was much higher in erythrocytes (90% inhibition) than in liver (20–40% inhibition). The increase in erythrocyte protoporphyrin and the decrease in both hemoglobin and hematocrit were not observed in mice exposed to lead under the above conditions.     

Determination of lead in samples of zooplankton,water, and sediments in a Mexican reservoir: Evidence for lead biomagnification in lower/intermediate trophic levels?

We have determined lead concentration of water, sediment, and zooplankton samples of El Niágara, a reservoir in Aguascalientes, Mexico. Our results include the first report of bioconcentration factor (BCF) obtained in an actual ecosystem (as opposed to the experimental setups in the laboratory) for a rotifer species; Asplanchna brigthwellii (BCF ca. 49 300). The BCF of this predatory zooplanktonic species (A. brigthwellii) are up to four times greater than those of two grazing zooplanktonic species (Daphnia similis and Moina micrura). In this contaminated reservoir that lacks fishes, Asplanchna, and Culex sp. together with ducks and other bigger invertebrates might represent the top predators. Our data suggest that biomagnification of lead through at least one trophic level can occur in freshwater systems. Biomagnification in A. brigthwellii might be explained in part by predation of this voracious predator on young of the herbivorous cladoceran, M. micrura. Our findings stand opposite to the current theoretical framework where lead biomagnification occurs only in lower trophic levels.     

Comparison of the in vitro binding characteristics of the β-carbolines harman and norharman in rat brain and liver and in bovine adrenal medulla

The in vitro binding of the naturally occurring -carbolines harman and norharman in their tritium-labelled forms to cell membranes from the rat brain and liver and from bovine adrenal medulla was investigated. Displacement of the specific [³H]harman binding in bovine adrenal medulla and rat liver by several -carbolines and monoamine oxidase (MAO) inhibitors revealed the pharmacological profile of a single, high-affinity binding site (K _D 4.92±0.43 nmol/l, B_max 8.47±0.17 pmol/mg protein; adrenal medulla) which corresponded to the active site of MAO type A (MAO-A). Similar characteristics have previously been found for brain tissue from rat, marmoset and pig. In order to determine the temperature dependence of the [³H]harman binding, the K _D and B_max values for rat cerebral cortex were calculated from the results of saturation experiments at 5 temperatures (range: 0°C–37°C). Whereas the B_max values under all conditions were – 4 pmol/mg protein, the K _D values, with increasing temperature, ranged from 3 nmol/l to 30 nmol/l. The calculated linear van't Hoff plot (-In K _D against 1/T) suggested an enthalpy-driven binding of [³H]harman to MAO-A.At least three different [³H]norharman-binding sites were detected. In the rat forebrain, 85% of the specific binding (at about 2 nmol/l of [³H]norharman) can be attributed to a MAO binding site of type B: the binding is displaceable, in nmol/l concentrations by the potent and selective MAO-B inhibitors MDL 72,974A, R(–)-deprenyl and pargyline and, in mol/l concentrations, by S(+)-deprenyl and the potent and selective MAO-A inhibitors clorgyline, harmine, harman, harmaline, brofaromine 5-F--methyltryptamine. After suppression of the MAO binding sites with 1 mol/l clorgyline and mol/l R(–)-deprenyl, a second binding site was found. However, the binding at this site was biphasically displaceable by harman and norharman (Hill-slopes about 0.5 and 0.6, curvilinear Rosenthal plots) suggesting the presence of negative co-operativity or of two binding sites (states). A similar clorgyline/R(–)-deprenyl resistent single (Hill-slopes of displacement by norharman, harman and 6-hydroxy--carboline about unity; linear Rosenthal plots) high affinity binding site (K _D 7.5±2 nmol/l, B_max 130±30 fmol/mg protein) was found in bovine adrenal medullary cell membranes. A third quite different clorgyline/R(–)-deprenyl resistent high-affinity (K _D14 nmol/l) and high-density (B_max 10–30 pmol/mg protein) binding site was detected in the liver. The specific binding at this site was not displaceable by harman or most other substituted -carbolines or by tetrahydro--carbolines, but was displaced by norharman and several newly synthesized 6-substituted aromatic -carbolines (e.g. F-, CH₃-, CH₃O-, HO-). The [³H]norharman binding site in the liver is certainly not identical with any of the binding sites for MAO-inhibitors, benzodiazepines or sigma receptor ligands and is slightly enriched in the microsomal (P₃) fraction whereas most of the specific [³H]harman binding was detected in the crude mitochondrial (P₂) fraction. Correspondence to: T. May at the above address     

Present drug-likeness filters in medicinal chemistry during the hit and lead optimization process: how far can they be simplified?

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GABAA ρ receptor mechanisms in the rat amygdala and its role in the modulation of fear and anxiety

Rationale  

Accumulating evidence for the presence of GABA_A ρ receptors within the amygdala which differ from other members of the GABA_A receptor family in both subunit composition and functional properties has been recently obtained.     

Source and impact of lead contamination on δ-aminolevulinic acid dehydratase activity in several marine bivalve species along the Gulf of Cadiz

Coastal areas and estuaries are particularly sensitive to metal contamination from anthropogenic sources and in the last few decades the study of space-time distribution and variation of metals has been extensively researched. The Gulf of Cadiz is no exception, with several rivers draining one of the largest concentrations of sulphide deposits in the world, the Iberian Pyrite Belt (IPB). Of these rivers, the Guadiana, one of the most important in the Iberian Peninsula, together with smaller rivers like the Tinto and Odiel, delivers a very high metal load to the adjacent coastal areas.The purpose of this work was to study the source and impact of lead (Pb) drained from historical or active mining areas in the IPB on the activity of a Pb inhibited enzyme (δ-aminolevulinic acid dehydratase, ALAD) in several bivalve species along the Gulf of Cadiz.Seven marine species (Chamelea gallina, Mactra corallina, Donax trunculus, Cerastoderma edule, Mytilus galloprovincialis, Scrobicularia plana and Crassostrea angulata) were collected at 12 sites from Mazagón, near the mouth of the rivers Tinto and Odiel (Spain), to Cacela Velha (Ria Formosa lagoon system, Portugal). Lead concentrations, ALAD activity and lead isotope ratios (²⁰⁶Pb/²⁰⁴Pb, ²⁰⁷Pb/²⁰⁴Pb and ²⁰⁸Pb/²⁰⁴Pb) were determined in the whole soft tissues.The highest Pb concentrations were determined in S. plana (3.50 ± 1.09 μg g⁻¹ Pb d.w.) and D. trunculus (1.95 ± 0.10 μg g⁻¹ Pb d.w.), while M. galloprovincialis and C. angulata showed the lowest Pb levels (<0.38 μg g⁻¹ Pb d.w.). In general, ALAD activity is negatively correlated with total Pb concentration. However this relationship is species dependent (e.g. linear for C. gallina ALAD = −0.36[Pb] + 0.79; r = 0.837; or exponential for M. galloprovincialis ALAD = 2.48e^−8.3[Pb]; r = 0.911). This indicates that ALAD activity has considerable potential as a biomarker of Pb and moreover, in marine bivalve species with different feeding habits. Lead isotope data showed significant seasonal and spatial changes in bivalve isotopic composition reflecting seasonal and geographic differences in bioaccumulation. Within the study area, Pb can be modelled as a mixing between geogenic Pb and mine-related, discharges of Pb from the IPB. For some sites at the mouth of the Guadiana River, the bivalves show contamination from other anthropogenic sources, such as leaded boat/aviation fuel and/or leaded paint. Finally, the study demonstrates convincingly the need to consider species-specific variation when using bivalve ALAD activity as a biomarker for Pb.     

Effects of historical lead–zinc mining on riffle-dwelling benthic fish and crayfish in the Big River of southeastern Missouri, USA

The Big River (BGR) drains much of the Old Lead Belt mining district (OLB) in southeastern Missouri, USA, which was historically among the largest producers of lead–zinc (Pb–Zn) ore in the world. We sampled benthic fish and crayfish in riffle habitats at eight sites in the BGR and conducted 56-day in situ exposures to the woodland crayfish (Orconectes hylas) and golden crayfish (Orconectes luteus) in cages at four sites affected to differing degrees by mining. Densities of fish and crayfish, physical habitat and water quality, and the survival and growth of caged crayfish were examined at sites with no known upstream mining activities (i.e., reference sites) and at sites downstream of mining areas (i.e., mining and downstream sites). Lead, zinc, and cadmium were analyzed in surface and pore water, sediment, detritus, fish, crayfish, and other benthic macro-invertebrates. Metals concentrations in all materials analyzed were greater at mining and downstream sites than at reference sites. Ten species of fish and four species of crayfish were collected. Fish and crayfish densities were significantly greater at reference than mining or downstream sites, and densities were greater at downstream than mining sites. Survival of caged crayfish was significantly lower at mining sites than reference sites; downstream sites were not tested. Chronic toxic-unit scores and sediment probable effects quotients indicated significant risk of toxicity to fish and crayfish, and metals concentrations in crayfish were sufficiently high to represent a risk to wildlife at mining and downstream sites. Collectively, the results provided direct evidence that metals associated with historical mining activities in the OLB continue to affect aquatic life in the BGR.     

Case reports of lead poisoning in dogs from the National Animal Poison Control Center and the Centre National D'Informations Toxicologiques, Veterinaires: anecdotes or reality?

This paper presents case reports of lead toxicoses from 2 major animal poison control centers in Europe and North America, gathered from 1985 through 1989. All results examined here involved cases assessed as "toxicosis" or "suspected toxicosis" by the National Animal Poison Control Center (NAPCC) or the Centre National d'Informations Toxicologiques Veterinaries (CNITV). 537 cases were reported to the NAPCC, most of them concerning dogs (59%). In France, most of the 362 cases involved cattle (57.2%). There was an increased number of cases reported during late summer and early fall, and a decreased number of cases in November and December, in both centers. Dogs intoxicated were predominantly young animals (60% were less than 2 years old). No sex difference was noted. Pure bred dogs appeared more often involved than mixed-breed ones, but the breed distribution closely resembles dog breed distribution in the US. The source of lead was usually unknown and, when information was available, paint seemed to be the most common cause of poisoning. Clinical signs reported to the animal poison control centers involved the CNS and GI tract. Results from the French and the American database showed similar trends. They are compared to data from veterinary clinics and veterinary colleges in the US and Australia. In each case, data are very similar to what was reported to the CNITV and the NAPCC. It is concluded that animal poison control centers databases can provide a useful tool for better knowledge of animal poisoning. They can also help identify unexpected toxicologic problems related to drug administration or pesticide use.     

O13 Computer Simulations as Alternatives in Education and in Research： the Sim-and cLabs-Programs and the BioSim Project

Teaching programs of the successfully used in practical While these programs originally ＂Virtual Physiology＂ series （ SimNerv, SimHeart, etc. ） are physiology and pharmacology courses all over the world. y might have been seen as only minor substitutes of the real labs it turned out in the course of teaching that they have major didactic advantages especially because the students are much more active in the virtual than in the animal labs. More recently, an advanced series of virtual computer labs （e. g. cLABs-Neuron） has been developed which also allows to do experiments that would be to difficult for most students to be physically carried out but can be realized in a virtual lab （http：//www. clabs, de/）.     

β-blockers and the risk of incident depression in the elderly

The hypothesis that β-blockers cause depression has been both confirmed and refuted in previous studies. However, in hardly any of these studies, depression was systematically and adequately assessed. The aim of this cohort study was to examine whether β-blockers, in general, highly lipid-soluble, nonselective, or serotonergic receptor-binding β-blockers, are associated with incident depression. Between 1993 and 2005, 5104 elderly persons were followed for incident depressions. Depressions were identified by regular interview and continuous monitoring of medical records. Cases were categorized as clinically relevant depressive symptoms or as depressive syndromes, the latter including Diagnostic and Statistical Manual of Mental Disorders-IV-defined depressive disorders. Pharmacies provided information on filled β-blockers. We used Cox regression with drug use as a time-dependent variable to analyze the data, adjusted for potential demographic covariates, activity of daily living, and (contra)indications for β-blockers. We found that use of β-blockers in general did not convey an increased risk of depressive symptoms (hazard ratio [HR], 0.76; 95% confidence interval [CI], 0.37-1.59) or depressive syndromes (HR, 0.99; 95% CI, 0.53-1.84). Highly lipid-soluble β-blockers, mostly propranolol in our study, were associated with depressive symptoms during the first 3 months of use (HR, 3.31; 95% CI, 1.03-10.6), but not with depressive syndromes. Nonselective or serotonergic receptor affinity was not associated with an increased risk of depressive symptoms or syndromes independent of high lipid solubility. We conclude that β-blockers in general do not convey an increased risk of depression. Lipophilic β-blockers are associated with an increased risk of depressive symptoms.     

The effects of lead exposure on placental NF-κB expression and the consequences for gestation

The present study was designed to investigate the toxicity of lead exposure on the placenta at different dosages and the relationship with placental expression of NF-κB. A total of 67 unrelated Han Chinese pregnant women and 108 Wistar rats were included in this study. The rats were randomly divided into four groups for consumption of water with or without 0.025% lead acetate during various gestational periods; blood samples and placenta were harvested for analysis. Blood lead content was determined by atomic absorption spectrophotometry. Placental NF-κB expression was evaluated by immunohistochemistry. Placental cytoarchitecture was examined by histopathology and electronic microscopy. Fetal body weight, body length and placental weight was significantly lower (p < 0.05) in the lead-exposed rats compared to controls. Maternal blood lead levels in the rats negatively correlated with placental weight (r = 0.652, p < 0.01). Rat placenta showed focal necrosis in the decidua with trophoblast degeneration and fibrin deposition. Mitochondria were swollen and decreased in number, rough endoplasmic reticula were distended and ribosomal number on membranes decreased. In the human placenta, we did not find abnormal cytoarchitecture. On the other hand, placental expression of NF-κB in lead-exposed rats was significantly higher than that in controls and the expression of NF-κB in human placenta was positively correlated with maternal blood lead levels (r = 0.663, p < 0.01). These findings suggest that lead exposure at various gestational periods produce varied effects, with NF-κB activation following lead exposure. Injury to cytoplasmic organelles may interfere with the nutrition and oxygen exchange between mother and fetus, which may be contribute to abnormal pregnancy outcomes.