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Toxicokinetics of 2,4,5-trichlorophenol and benzo(a)pyrene in the clam Pisidium amnicum: effects of seasonal temperatures and trematode parasites.

Several biotic and abiotic stress factors may affect aquatic organisms simultaneously. However, not much is known about the effects of, e.g., low temperatures and parasite infections on the toxicokinetics of organic hydrophobic chemicals. Here we studied the accumulation and depuration of [(14)C]2,4,5-trichlorophenol (TCP) and [(3)H]benzo(a)pyrene (BaP) in the sediment-dwelling freshwater clam Pisidium amnicum. Experiments were made in October (+15 degrees C), April (+4 degrees C), and July (+15 degrees C) both with uninfected clams and clams infected with Bunodera luciopercae (Trematoda). The accumulation rate for both chemicals was slower at 4 degrees C than at 15 degrees C. The depuration of TCP was biphasic, and the slowest depuration occurred at 4 degrees C. For BaP, the depuration was very slow and monophasic at all temperatures. The highest BCFs for both chemicals were found in July at 15 degrees C. Surprisingly, the BCFs for TCP were higher in April at 4 degrees C than in October at 15 degrees C. For BaP, no steady-state was reached in April. Differences in chemical toxicokinetics between the infected and uninfected clams were only minor. However, for both chemicals a trend of slightly lower BCFs in the infected clams was found. In conclusion, low temperatures modify the toxicokinetics of organic chemicals in P. amnicum and the effects depend on hydrophobicity of the chemical. The effects of parasites on toxicokinetics seem to be small.

Algorithms↗

Degradation of prochloraz and 2,4,6-trichlorophenol by environmental bacterial strains.

Eight bacterial isolates from enrichment with 2,4,6-trichlorophenol (TCP) as sole carbon source were tested for their potential to degrade prochloraz. None of them could grow on prochloraz. Strain C964, identified as Aureobacterium sp., effectively reduced the fungitoxic activity of prochloraz in a bioassay and degradation was confirmed by HPLC. Two other isolates, strain C611 and C961, using TCP as a carbon source, belong to the beta subclass of the proteobacteria and presumely degrade TCP via 2,4-dichlorohydroquinone and hydroxyhydroquinone as indicated by oxygen-consumption tests.

Bacteria↗

Transformation of 2,4,6-trichlorophenol by free and immobilized fungal laccase.

Laccase from the white rot fungus Coriolus versicolor was immobilized on Celite R-637 by covalent binding with glutaraldehyde. After a sharp primary decline in activity (up to 50%), the retained enzyme activity was stable over a storage period of 33 days at 4 degrees C. A comparative study of soluble and immobilized laccases revealed the increased resistance of immobilized enzyme to the unfavourable effects of alkaline pH, high temperature and the action of inhibitors. A combination of these properties of immobilized laccase resulted in the ability to oxidize 2,4,6-trichlorophenol (2,4,6-TCP) at 50 degrees C at pH 7.0. The reactions of soluble and immobilized laccase with 2,4,6-TCP were examined in the presence and absence of redox mediators. 3,5-Dichlorocatechol, 2,6-dichloro-1,4-benzoquinone and 2,6-dichloro-1,4-hydroquinone were found to be the primary products of 2,4,6-TCP oxidation by laccase; oligo- and polymeric compounds were also found.

Basidiomycota↗

Clinical, radiological and histological study of the failure of cervical interbody fusions with bone substitutes.

Few histological studies on bone substitutes in human cervical spine are available and the biological processes of bone substitutes are not well documented. The authors studied four failure cases of cervical interbody fusion: two cases with hydroxyapatite (HA), one case with beta-tricalcium phosphate ceramic (beta-TCP) and one case with xenograft (bovine bone). Clinical data showed that all the patients experienced neck pain with or without numbness of upper extremity due to fusion failure. Successful fusions were achieved after the salvage surgeries in which autograft were used. Radiographs showed that radiolucent lines were present in all cases. Two HA substitutes fractured without complications. One of them sank into the vertebral body. Some small beta-TCP fragments were found under the microscope. Histological study demonstrated only a few newly formed bones at the interface of the substitutes. The fragments of HA were encapsulated by fibrous tissue. The degradation process and bone regeneration were more active in beta-TCP than in HA. The intertrabecular spaces of bovine bone were filled with fibrous tissue. The results suggest that a porous calcium phosphate ceramic with special design might assure bone ingrowth and meet the mechanical requirements in cervical interbody fusion. The complications of these materials in the cervical spine should be highlighted.

Adult↗

[Novel calcium phosphate formula for filling bone defects. Initial in vivo long-term results].

BACKGROUND: Up to now hydroxyapatite (HA) and beta-tricalciumphosphate (beta-TCP) ceramics have been routinely sintered at temperatures between 1100 degrees and 1500 degrees C. Our new calcium ceramic is fabricated by a sol-gel process at 200 degrees C. The aim of this investigation was to test the biodegradation of and the induction of bone formation by this material. MATERIAL AND METHODS: Eighteen 1-year-old Goettingen minipigs were divided into three groups. Critical size defects (>5 cm(3)) in the mandible were treated differently in all three animals (group 1: filling with 40% beta-TCP plus 60% HA, group 2: pure HA was applied, group 3 served as controls: only gelatinous material was given). Macroscopic and microscopic investigations of the former defects were made 8 months postoperatively. RESULTS. In groups 1 and 2 biodegradation of more than 93% of the new calcium phosphate formula was found 8 months postoperatively and considered to be very good. No difference was observed between pure HA (group 2) and the combination of HA and beta-TCP (group 1). In both groups complete bone formation was seen macroscopically in the former defects. In the control group only incomplete bone formation with 48.4% of the defect area was noted. This difference was significant ( p<0.001). DISCUSSION: The new calcium phosphate formula made by a sol-gel method at 120 degrees C seems to be suitable for filling bone defects and is of interest for orthopedic surgery, traumatology, craniomaxillofacial surgery, and dentistry.

Absorbable Implants↗

Biosynthesis of poly-beta-hydroxyalkanoates by Sphingopyxis chilensis S37 and Wautersia sp. PZK cultured in cellulose pulp mill effluents containing 2,4,6-trichlorophenol.

Poly-beta-hydroxyalkanoates (PHA) polymer is synthesized by different bacterial species. There has been considerable interest in the development and production of biodegradable polymers; however, the high cost of PHA production has restricted its applications. Kraft cellulose industry effluents containing 2,4,6-trichlorophenol (10 or 20 microg ml(-1)) were used by the bacteria Sphingopyxis chilensis S37 and Wautersia sp. PZK to synthesize PHA. In this condition, S. chilensis S37 was able to grow and degrade 2,4,6-trichlorophenol (ca. 60%) and 80% of these cells accumulated PHA. Wautersia PZK completely degraded 2,4,6-TCP and more than 90% of the cells accumulated PHA in 72 h. The PHA detection was performed by flow cytometry and polyester composition was characterized by gas chromatography-mass spectroscopy (GC-MS), indicating that these polymers are made by 3-hydroxybutyric acid and 3-hydroxyhexadecanoic acid for S37 and PZK strains, respectively. Results demonstrated that strains' growth and PHA production and composition are not modified in cellulose effluents with or without 2,4,6-TCP (10-20 microg ml(-1)). Therefore, our results indicate that S. chilensis S37 and Wautersia sp. PZK are able to degrade a toxic compound such as a 2,4,6-TCP and simultaneously produce a valuable biopolymer using low-value substrates.

3-Hydroxybutyric Acid↗

Effects of TiO2, ZrO2 and Al2O3 dopants on the compressive strength of tricalcium phosphate.

Tricalcium phosphate (TCP) powders synthesised using the Ca(NO3)2 and Ca(OH)2 routes were doped with TiO2, ZrO2 and Al2O3 in order to increase their compressive strength. An ultimate compressive strength (UCS) of 255 +/- 6 MPa was achieved for approximately 10 vol% TiO2 doping compared to 30 +/- 3 MPa for an un-doped control processed and tested in the same manner. Higher levels of TiO2 doping resulted in smaller increases in UCS with 30 and 50 vol% achieving 213 +/- 9 and 178 +/- 15 MPa, respectively. Very small amounts of Al2O3 doping (< 0.5 vol%) also resulted in a stronger materials. However, under the processing conditions employed, higher levels of Al2O3 and ZrO2 doping resulted in no beneficial effect on the UCS. Polyvinyl alcohol (PVA) was used as binding agent to facilitate processing. As expected, higher levels of PVA were associated with smaller increases in UCS. Powders synthesised using the Ca(OH)2 route had smaller particle size and resulted in larger increases in UCS compared to the Ca(NO3)2-synthesised powders. Although some powders contained alpha and beta-TCP phases, no other calcium phosphate, CaO, CaTiO3 or CaZrO3 phases were detected. In conclusion, a significant increase in the UCS of TCP was achieved by doping with approximately 10 vol% TiO2 which is expected to have little or no effect on the bioactivity or bioresorbability of the material.

Aluminum Oxide↗

A novel bioactive porous bredigite (Ca7MgSi4O16) scaffold with biomimetic apatite layer for bone tissue engineering.

The aim of this study was to develop a novel bioactive, degradable and cytocompatible bredigite (Ca(7)MgSi(4)O(16)) scaffold with biomimetic apatite layer for bone tissue engineering. Porous bredigite scaffolds were prepared using polymer sponge method. The bredigite scaffolds with biomimetic apatite layer (BTAP) were obtained by soaking bredigite scaffolds in simulated body fluid (SBF) for 10 days. The porosity and in vitro degradability of BTAP scaffolds were investigated. In addition, osteoblast-like cell morphology, proliferation and differentiation on BTAP scaffolds were evaluated and compared with beta-tricalcium phosphate (beta-TCP) scaffolds. The results showed that BTAP scaffolds possessed 90% of porosity. The degradation of BTAP scaffolds was comparable to that of beta-TCP scaffolds. Cells on BTAP scaffolds spread well and presented a higher proliferation rate and differentiation level as compared with those on beta-TCP scaffolds. Our results indicated that BTAP scaffolds were degradable and possessed the function to enhance cell proliferation and differentiation, and might be used as bone tissue engineering materials.

Absorbable Implants↗

The mechanical properties of calcium phospate ceramics modified by collagen coating and populated by osteoblasts.

Bioceramics containing hydroxyapatite (HA), tricalcium phosphate (TCP) or composites which combine the best properties of both materials are among the principal candidates for bone replacement grafts. In this study we have investigated the mechanical strength of HA, TCP and composites of the two in the ratios 75;25 (H75), 50:50 (H50) and 25:75 (H25). The strength of each material was investigated in the presence and absence of collagen coating, and the influence of osteoblast culture for up to 28 days on strength was determined. TCP, H25 and H75 were significantly weakened by collagen coating, the strengths of the other materials were either not affected (HA) or increased (H50). Culture with osteoblasts significantly increased the strength of uncoated HA and H50, but this effect was not observed when the materials were coated with collagen. Our results indicate that ceramic composition affects the interactions between collagen coating, culture with osteoblasts and mechanical strength of the material. Although collagen coating has been found to increase the proliferation of osteoblasts into these ceramic materials, it may be necessary to stabilise and optimise the coating process to minimise effects on mechanical strength.

Animals↗

Multiple mode of binding of phencyclidines: high affinity association between phencyclidine receptors in rat brain and a monovalent ion-sensitive polypeptide.

Two populations of phencyclidine (PCP) binding sites are shown to exist in the rat brain: a high-affinity monovalent ion-sensitive site (Kd of 10-14 nM for [3H]TCP, [3H]N-[1-(2-thienyl)cyclohexyl]piperidine), which exists in both the frontal cortex and the hippocampus, and a lower affinity site (Kd of 80-130 nM for [3H]TCP) which is found in the hippocampus but not in the frontal cortex. The nature of the interactions between the ion-binding sites and the high affinity PCP receptors depend on both ligand structure (PCP or TCP) and the ion involved (K' or Na'). The high-affinity sites are associated with an Mr 90,000 polypeptide whose labeling by [3H]azido phencyclidine is selectively inhibited by monovalent ions.

Animals↗

Presence of sigma and phencyclidine (PCP)-like receptors in membrane preparations of bovine adrenal medulla.

Receptor binding studies with [3H]-(+)SKF-10047 were carried out to characterize the putative sigma (sigma) and phencyclidine (PCP) receptors in membrane preparations of bovine adrenal medulla. Specific binding of the radiolabelled compound was observed after incubation with the membrane preparation at 37 degrees, the equilibrium being reached at 20 min and the maximal binding being observed with 0.6 mg/ml protein. Saturation binding studies were performed at equilibrium (30 min at 37 degrees with 0.5 mg/ml of membrane protein) in the presence of haloperidol (1 microM) or 1-[1-(2-thienyl) cyclohexyl] piperidine (TCP; 0.2 microM) to block sigma or PCP receptors, respectively. The binding of [3H]-(+)SKF-10047 was characterized by two distinct components. A high affinity binding site (haloperidol sensitive) had an apparent KD of 8.3 nM and a Bmax of 67 pmol/g protein. A lower affinity binding site (TCP sensitive) had an apparent KD of 32.7 nM and a Bmax of 83 pmol/g protein. The drug specificity of the high affinity binding site resembled that of the putative sigma receptor, being potently inhibited by haloperidol and pentazocine. The binding pharmacology of the low affinity site resembled that of the phencyclidine receptor, being potently displaced by TCP and PCP. The binding of [3H]-(+)SKF-10047 to both receptors showed marked stereoselectivity for the dextrorotatory (+) isomer of SKF-10047 and was insensitive to the receptor specific opioid ligands DAGO (mu), DSLET (delta) and U-69593 (kappa). These data indicate that bovine adrenal medulla contains sigma and PCP-like receptors.

Adrenal Medulla↗

A gas chromatographic procedure for separation and quantitation of the enantiomers of the antidepressant tranylcypromine.

A novel assay procedure has been developed that allows for the separation and quantification of the enantiomers of the monoamine oxidase inhibitor tranylcypromine (TCP) in brain and liver of rats. The analytical method involves extraction of the drug from rat tissue with an organic solvent. TCP is then derivatized with S-(-)-N-(trifluoroacetyl)-prolyl chloride to allow gas chromatographic analysis of the resulting diastereoisomers. Conditions for analysis by a gas chromatograph equipped with a nitrogen-phosphorus detector and a capillary column are described. The method has been applied to the separation and quantification of the enantiomers of TCP in samples of brain and liver of rats that had been injected with this drug alone and after pretreatment with iprindole, a drug known to block aromatic ring hydroxylation.

Animals↗

GABAergic neurons in human spinal cord cultures: a computer-aided analysis of normal and thienyl-phencyclidine-treated cells.

gamma-Aminobutyric acid (GABA)-containing neurons were studied in dissociated cell cultures of human spinal cords from 6-10-week-old fetuses using immunohistochemistry with anti-GABA antibodies. Light microscopy showed two types of immunoreactive (IR) neurons: (1) IR neurons with short neuritic processes remaining near the cell body (small neuritic tree neurons); and (2) IR neurons with long neuritic processes extending far from the cell body (large neuritic tree neurons). Both types were studied at different ages in vitro, in control and in thienyl phencyclidine (TCP)-treated cultures by means of computer reconstructions and morphometric parameters. A discriminant analysis permitted the recognition of three populations: whatever the age, the control and TCP-treated neurons with small neuritic trees were not discriminated from each other and were considered to be one population whereas the 98 DIV control and both 21 DIV and 98 DIV TCP-treated cells with large neuritic trees were clearly separated from each other and from the small cell population. In all models, an astrocytic labeling, weaker than that of the neurons, was observed. The nature of these neurons (probably interneurons) intrinsic to the spinal cord is discussed in view of previous findings concerning the anatomical distribution and organization of the GABAergic system in the spinal cord.

Cells, Cultured↗

Synthesis of mouse t complex proteins during haploid stages of spermatogenesis.

We have analyzed the expression, through spermiogenesis, of a series of testicular cell polypeptides encoded by genes within the mouse t complex. Two of these polypeptides, TCP-3 and TCP-7, are synthesized in a testes-specific manner with highest levels of expression during haploid stages of spermatogenesis. A third, TCP-1, is also expressed at highest levels in haploid cells, and expression of this polypeptide continues until the last residual body stage of spermiogenesis. The genes that encode these polypeptides have been correlated with the t phenotype of transmission ratio distortion.

Animals↗

Effects of amygdaloid kindling on NMDA receptor function and regulation.

These studies were conducted to determine whether amygdaloid kindling results in the long-term alteration of NMDA receptors which could explain the persistent reduction in seizure threshold seen in this phenomenon. NMDA-induced [3H]norepinephrine (NE) release, NMDA-sensitive L-[3H]glutamate binding, and NMDA and glycine-enhanced [3H]TCP binding were measured in brain tissue from kindled rats and nonstimulated control rats 3 to 6 weeks after the last seizure. There was no difference in the ability of NMDA to induce [3H]NE release from kindled or control slices of amygdala or hippocampus. There was also no difference in the ability of phencyclidine (PCP) or Mg2+ to inhibit [3H]NE release induced by 100 microM NMDA. Equilibrium saturation experiments of NMDA-sensitive L-[3H]glutamate binding revealed no differences in KD or Bmax values between control and kindled cortex, amygdala, and hippocampus. The Ki values for NMDA displacement of L-[3H]glutamate binding also did not differ in kindled tissue. NMDA-enhanced [3H]TCP binding was similar in cortex, amygdala, and hippocampus of kindled and control tissues. Finally, glycine-enhanced [3H]TCP binding was not different in control or kindled tissues. These studies suggest that the NMDA recognition site and the modulation of the NMDA receptor/ion channel complex by magnesium, PCP, and glycine are not altered several weeks after the last seizure. Even though NMDA-mediated electrophysiological responses are reportedly enhanced in kindled tissue at that time, the mechanism(s) underlying the enhancement remains to be determined.

Amygdala↗

Solubilization of phencyclidine receptors from rat cerebral cortex in an active ligand binding state.

A phencyclidine (PCP) receptor binding site has been solubilized in an active ligand-binding state from rat cerebral cortical membranes with sodium deoxycholate. Optimal receptor solubilization occurs at a detergent/protein ratio of 0.5 (w/w); for 5 mg protein/ml solubilized with 0.25% sodium deoxycholate, about 60% of the protein and 25% of the receptor is solubilized. Specific binding of either [3H]-N-[1-(2-thienyl)cyclohexyl]piperidine ([3H]TCP) or [3H]MK-801 is measurable by filtration through Sephadex G-50 columns or glass fiber filters; more than 60% of the binding activity is stable after 48 h at 4 degrees C. In the presence of detergent, [3H]TCP binding exhibits a Kd of 250 nM, a Bmax of 0.56 pmol/mg protein, and a pharmacological profile consistent with that of the membrane-bound PCP receptor, although most drugs bind with affinities 2 to 8 fold lower than in membranes. Upon reduction of detergent concentration, binding parameters approximate those for the membrane-bound receptor ([3H]TCP binding: Kd = 48 nM, Bmax = 1.13 pmol/mg protein).

Animals↗

Pseudoallosteric modulation by (+)-MK801 of NMDA-coupled phencyclidine binding sites.

Two high affinity phencyclidine (PCP) binding sites, labelled by [3H] 1-[1-(2-thienyl)cyclohexyl]piperidine ([3H]TCP), have been identified in guinea pig brain, with one site coupled to the N-methyl-D-aspartate (NMDA) receptor (site 1) and the other site associated with the dopamine reuptake carrier complex (site 2). In this study, PCP enhanced the dissociation of [3H]TCP from PCP site 1 and site 2, while (+)-MK801 only enhanced dissociation of [3H]TCP from PCP site 1. Although additional studies will be required to determine the exact mechanism(s) of these effects, these data demonstrate that the interactions of PCP with both site 1 and site 2 are more complex than previously appreciated.

Allosteric Regulation↗

The effects of rubidium, caesium and quinine on 5-HT-mediated behaviour in rat and mouse--1. Rubidium.

The administration of TCP (15 mg/kg, i.p.) to rats pretreated with either intraperitoneal RbCl (3 mmol/kg, twice daily for 5 days) or dietary RbCl (30 mmol/kg diet, for 14 days), resulted in the complete 5-HT behavioural syndrome. Pretreatment with p-chlorophenylalanine (i.p. 300 mg/kg x2) or (-)-propranolol (20 mg/kg, i.p.), pindolol (4 mg/kg, i.p.) and ritanserin (0.4 mg/kg, s.c.) prevented the occurrence of the 5-HT syndrome, produced by dietary RbCl plus TCP. Intraperitoneal administration of RbCl had no effect upon the 5-HT behavioural syndrome, produced by 8-OH-DPAT (0.5 mg/kg, s.c.) or 5-MeODMT (2 mg/kg, i.p.) but enhanced the 5-HT syndrome produced by quipazine (20 mg/kg, i.p.), DOI (8 mg/kg, s.c.), p-chloramphetamine (4 mg/kg, i.p.) or by TCP plus L-tryptophan (50 mg/kg, i.p.) in rats. Dietary administration of RbCl resulted in the enhancement of the 5-HT2-mediated head-twitches in the mouse and the attenuation of hypothermia in the mouse, induced by 8-OH-DPAT (0.5 mg/kg, s.c.). The accumulation of 5-HT (after inhibition of monoamine oxidase) and the rate of synthesis of 5-HT in the whole brain (minus cerebellum) were enhanced by dietary and intraperitoneal administration of RbCl, respectively. The effects of lithium and rubidium, respectively, on 5HT function in brain are compared.

8-Hydroxy-2-(di-n-propylamino)tetralin↗