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Preliminary beta-tricalcium phosphate coating prepared by discharging in a modified body fluid enhances collagen immobilization onto titanium.

Because of its excellent scaffold properties toward bone cells, collagen has been recognized as a promising extracellular matrix protein for surface modification of titanium implants. Hydroxyapatite (HA) coatings have been investigated as a preliminary coating for collagen immobilization on titanium implants. However, the composition of HA-collagen is recognized as being difficult, and while many studies have suggested that biodegradable beta-tricalcium phosphate (beta-TCP) has better osteoconductivity than HA, the efficiency of preliminary beta-TCP coating for collagen immobilization on titanium surfaces has yet to be evaluated. This investigation aimed to evaluate the applicability of HA and beta-TCP coatings, prepared by discharging in modified body fluids, as preliminary collagen coatings. To increase collagen induction on preliminary HA and beta-TCP coatings, we used a new cathodic polarization method. X-ray photoelectron spectroscopy revealed that the bonding strength between the collagen NH(+) amino groups of collagen and phosphate (PO(4) (3-)) was greater on the beta-TCP coating than the HA coating. The preliminary beta-TCP coating was tightly crosslinked with RCOO(-) carboxyl groups of the collagen molecules and showed high cellular responses, even in the early stage of cell cultivation. Thus, this coating was found to be more effective than HA as a preliminary coating for collagen immobilization on titanium implants.

Animals↗

Polypyrrole thin films formed by admicellar polymerization support the osteogenic differentiation of mesenchymal stem cells.

The objective of this study was to evaluate the attachment, proliferation, and differentiation of rat mesenchymal stem cells (MSC) toward the osteoblastic phenotype seeded on polypyrrole (PPy) thin films made by admicellar polymerization. Three different concentrations of pyrrole (Py) monomer (20, 35, and 50 x 10(-3) M) were used with the PPy films deposited on tissue culture polystyrene dishes (TCP). Regular TCP dishes and PPy polymerized on TCP by chemical polymerization without surfactant using 5 x 10(-3) M Py, were used as controls. Rat MSC were seeded on these surfaces and cultured for up to 20 d in osteogenic media. Surface topography was characterized by atomic force microscopy, X-ray photoelectron spectroscopy, and static contact angle. Cell attachment, proliferation, alkaline phosphatase (ALP) activity, and calcium content were measured to evaluate the ability of MSC to adhere and differentiate on PPy-coated TCP. Increased monomer concentrations resulted in PPy films of increased thickness and surface roughness. PPy films generated by different monomer concentrations induced drastically different cellular events. A wide spectrum of cell attachment characteristics (from excellent cell attachment to the complete inability to adhere) were obtained by varying the monomer concentration from 20 m to 50 x 10(-3) M. In particular the 20 x 10(-3) M PPy thin films demonstrated superior induction of MSC osteogenicity, which was comparable to standard TCP dishes, unlike PPy films of similar thickness prepared by chemical polymerization without surfactant. Adhesion of mesenchymal stem cells on tissue culture plates (TCP) coated with polypyrrole thin films made by admicellar polymerization.

Alkaline Phosphatase↗

The aquatic fate of triclopyr in whole-pond treatments.

The aquatic fate of the triethylamine salt formulation of triclopyr (3,5,6-trichloro-2-pyridinyloxyacetic acid) was determined in whole-pond applications in closed (no water exchange) systems in California, Missouri and Texas in two studies conducted in 1995 and 1996. These studies determined dissipation rates of triclopyr and its principal metabolites, 3,5,6-trichloropyridinol (TCP) and 3,5,6-trichloro-2-methoxypridine (TMP) in water, sediment and finfish. Ponds at each site containing a healthy biological community were treated at 2.5 mg AE litre-1 triclopyr. Water and sediment samples were collected through 12 weeks post-treatment, and non-target animals were collected through 4 weeks post-treatment. Dissipation rates for triclopyr, TCP and TMP were similar at each of the study sites, despite differences in weather, water quality, biotic community, light transmission and geographic location. Half-lives of triclopyr in water ranged from 5.9 to 7.5 days, while those of TCP and TMP ranged from 4 to 8.8 and 4 to 10 days, respectively. Levels of triclopyr and TCP declined in sediments at half-lives ranging from 2.8 to 4.6 days and 3.8 to 13.3 days, respectively. No TMP was detected in sediment. Triclopyr and TCP cleared from fish in relation to concentrations found in the water column. TMP levels in fish were generally an order of magnitude higher than levels of triclopyr and TCP, particularly in the visceral portion of the animals. No adverse effects on water quality or on the non-target biotic community were found following triclopyr applications. Results of these studies were comparable to those of triclopyr dissipation studies conducted in reservoirs, lakes and riverine systems in Georgia, Florida, Minnesota and Washington, indicating that the degradation and dissipation of triclopyr and its metabolites are similar in representative systems throughout the USA.

Animals↗

[3H]1-[2-(2-thienyl)cyclohexyl]piperidine labels two high-affinity binding sites in human cortex: further evidence for phencyclidine binding sites associated with the biogenic amine reuptake complex.

Previous work demonstrated two high-affinity PCP binding sites in guinea pig brain labeled by [3H]TCP (1-(1-[2-thienyl]cyclohexyl)piperidine): site 1 (N-methyl-D-aspartate [NMDA]-associated) and site 2 (dopamine-reuptake complex associated). The present study examined brain membranes prepared from various species, including human, for the presence of site 2, defined as binding in the presence of (+)-5-methyl-10,11-dihydro-5H-dibenzo [a, d]cyclohepten-5,10-imine maleate ((+)-MK801) minus binding in the presence of 10 microM TCP (nonspecific binding). Studies were conducted in absence of sodium which was found to be inhibitory to [3H]TCP binding. The results demonstrated detectable levels of site 2 in brain membranes of guinea pig, rabbit, pig, mouse, sheep, and human but not in the rat or chicken. Using human cortical membranes, site 2 was the predominant binding site. Detailed studies conducted with human cortical tissue showed that high-affinity dopamine (1-[2- [bis(4-fluorophenyl)-methoxy]ethyl]-4-(3-phenylpropyl)piperazine (GBR12909)], [1,2]benzo(b)thiophenylcyclo-hexylpiperidine (BTCP), and serotonin (fluoxetine) uptake inhibitors produced a wash-resistant inhibition of [3H]TCP binding to site 2, but not site 1. Preincubation of guinea pig brain membranes with BTCP was shown to produce an increase in the dissociation rate of [3H]TCP from PCP site 2. Structure activity studies with various uptake inhibitors showed that GBR12909, benztropine, fluoxetine, and BTCP have higher affinity for site 2 than for site 1. (+)-MK801, ketamine, and tiletamine were very selective for site 1, whereas dexoxadrol and TCP were moderately selective for site 1. These results suggest that human cortex possesses high-affinity PCP binding sites associated with biogenic reuptake binding sites, and that guinea pig brain, but not rat brain, may be an appropriate animal model for studying PCP site 2 in human brain.

Animals↗

Subchronic inhalation toxicity of 1,1,1,3-tetrachloropropane in rats.

The purpose of this study was to evaluate the inhalation toxicity of 1,1,1,3-tetrachloropropane (TCP), an intermediate in the production of chlorinated silicone fluids. Male and female Sprague-Dawley rats were exposed 6 hr/day, 5 days/week, for 90 days to TCP at concentrations of 0, 25, 75, or 225 ppm (Phase I study) and to 0, 1, 5, or 10 ppm (Phase II study). Phase II of the study was conducted because a no-observed-effect level was not achieved in Phase I. No animals died during the study. Clinical signs of toxicity included oral, nasal, and/or ocular discharge. No statistically significant differences were observed in either body weights or food consumption between exposed and control animals. Clinical pathology did not indicate any treatment-related effects. Absolute and relative liver and kidney weights were increased in male and female rats exposed to 225 ppm TCP, and heart weights were increased in male rats exposed to 225 ppm TCP. The liver and heart weight changes were supported by the findings of microscopic lesions in these organs. These lesions consisted of multifocal/focal myofiber degeneration necrosis with adjacent chronic myocarditis in the heart and multifocal single-cell necrosis in the liver parenchyma. The liver lesions had essentially resolved at the end of a 28-day recovery period but the heart lesions were still present in male rats in the recovery group exposed to 225 ppm TCP. No treatment-related effects were observed in animals exposed to 1, 5, or 10 ppm TCP.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗

Does prostacyclin mediate alpha-adrenergic induced hypotension?

We have extended our previous observations on the effect of tranylcypromine (TCP) on intraocular pressure (IOP), following topical administration of catecholamines is normal and chemically denervated rabbit eyes. In normal eyes, TCP inhibits the hypotensive phase after topical norepinephrine (nE); less after epinephrine (E); and not at all after isoproterenol. In denervated eyes, the inhibitory effect of TCP on the hypotensive phase of nE and E is enhanced. Phenoxybenzamine (PBA), but not timolol maleate or indomethacin, blocks the effect of TCP on alpha-adrenergic induced hypotension. Prostacyclin-like activity was estimated by bioassay using ADP induced rat platelet aggregation. This activity is significantly reduced in the primary aqueous and in the iris after TCP but it is significantly increased in pooled data of aqueous samples after topical nE. We conclude that the inhibitory effect of TCP on the hypotensive phase after topical E and nE is the result of inhibition of prostacyclin synthesis and not of MAO inhibition nor blockade of alpha-receptors. It is possible that the normal production of prostacyclin by the iris and ciliary body maintains (lower) basal levels of IOP.

Animals↗

Specific action of tranylcypromine to precipitate barbital withdrawal convulsions.

This research was designed to determine whether the convulsion-eliciting action of tranylcypromine (TCP) during bartibal (B) withdrawal was specific to physical dependence on B, and to compare the findings with the action of pentetrazol (PTZ). Challenge with 15-20 mg/kg IP TCP at 48 or B withdrawal resulted in the elicitation of clonic-tonic convulsion (CTC) in all rats (n = 6) within 10 min. Another challenge with 5-20 mg/kg TCP led to the dose-related precipitation and intensification of CTC. The CTC-inducing action of TCP was relatively reduced as the B withdrawal signs were gradually mitigated. In other words, when challenge with the drug was made at 72h of B withdrawal, the time of CTC onset was prolonged, and the incidence was reduced to 50% in parallel with abolition of the other withdrawal signs. A challenge at 120 h of B withdrawal when the vital signs had almost recovered to prewithdrawal level failed to induce even the prodromal signs of convulsion. In all rats exhibiting only mild to moderate withdrawal signs (such as hyperirritability, hyper-reflexia, anorexia, weight loss), 40 mg/kg TCP was required to induce CTC during B withdrawal, which was twice the dose required in severely dependent rats. Other monoamine oxidase inhibitors, i.e., pargyline, iproniazid, and clorgyline, elicited no CTC during B withdrawal. Methamphetamine was without effect on B withdrawal convulsions. From these findings, the convulsive threshold for TCP during B withdrawal proved well correlated to the grade of B-dependence and the duration of B-withdrawal signs.

Animals↗

Role of monoamines in behavior of reserpinized rats given tranylcypromine stereoisomers.

The effects of tranylcypromine stereoisomers [(+)-TCP and (-)-TCP] on behavior were studied in reserpinized rats. Rats given reserpine showed a behavioral syndrome characterized by hypoactivity. Administration of (+)-TCP to reserpinized rats led to a dose-dependent increase in their locomotor activity. At a dose of 15 mg/kg, (+)-TCP produced activation, stereotypy and hyperstimulation in reserpinized rats. In contrast, (-)-TCP failed to influenced the behavior of reserpinized rats reliably. Pharmacological studies using agonists and antagonists of monoaminergic functions showed the actions of (+)-TCP to depend on presynaptic serotonergic and catecholaminergic functions. The results provide another example of stereoselective effects of tranylcypromine stereoisomers on behavior mediated mainly by monoaminergic neurotransmission.

Animals↗

Effects of tranylcypromine enantiomers on monoamine uptake and release and imipramine binding.

Studies were carried out in vitro to determine effects of tranylcypromine enantiomers ([+]- and [-]-TCP) on uptake and release of 5-HT, DA and NA in rat synaptosomes and on imipramine binding to rabbit platelets. (+)-TCP was more potent than (-)-TCP as inhibitor of 5-HT uptake and imipramine binding, whereas (-)-TCP was more potent than (+)-TCP as inhibitor of DA and NA uptake. The enantiomers differed only slightly in their effects on monoamine release. The findings agree with previous reports on the stereo-selectivity of monoaminergic mechanisms toward TCP enantiomers, and support the notion that the 5-HT uptake site may be associated with the imipramine binding site.

Amines↗

[The transverse coloplasty pouch after low anterior resection: early postoperative results].

INTRODUCTION: A colon J pouch (CJP) still represents the standard rectal reservoir after low anterior resection. Though the CJP shows favourable early functional results, pouch evacuation problems tend to occur in the long term. The transverse coloplasty pouch (TCP), developed by our group allows comparable early functional results while avoiding evacuation problems. We report our experience with the TCP at the University Hospital of Heidelberg, Germany, and examine the risk of anastomotic leaks with this technique. METHODS: Between 1 October 2001 and 31 May 2003, 201 patients with rectal tumours underwent resection. Eighty-two patients with formation of TCP were enrolled in the study. RESULTS: During the creation of the TCP, no technical problems occurred, and the overall morbidity was 28%, including anastomotic leaks in seven patients (8.5%) and bleeding in two. The reoperation rate was 8.5%. An association between postoperative morbidity and preoperative radiation therapy could not be established. The hospital mortality rate was 3.6%. CONCLUSIONS: The use of TCP is a safe procedure which has gained worldwide acceptance in a short time, representing a technically straightforward procedure. Independently of patient size, habitus, and bulkiness of the colon, a TCP can always be performed after low rectal resection.

Adult↗

Arabidopsis glucosyltransferases with activities toward both endogenous and xenobiotic substrates.

Arabidopsis thaliana Heynh. harbors UDP-glucose-dependent glucosyltransferase (UGT; EC 2.4.1.-) activities that are able to glucosylate xenobiotic substrates as a crucial step in their detoxification, similar to other plants. However, it has remained elusive whether side-activities of UGTs acting on endogenous substrates could account for that property. Therefore, seven recombinantly expressed A. thaliana enzymes were tested using the phytotoxic xenobiotic model compound 2,4,5-trichlorophenol (TCP) as a substrate. The enzymes were selected from the large Arabidopsis UGT gene family because their previously identified putative endogenous substrates comprised both carboxylic acid, and phenolic and aliphatic hydroxyl moieties as biochemical targets. In addition, UGT75D1, which was shown to accept the endogenous flavonoid kaempferol as a substrate, was included. All enzymes tested, except the sterol-conjugating UGT80A2, glucosylated TCP as a parallel activity. The K(m) values for TCP ranged from 0.059 to 1.25 mM. When tested at saturating concentrations of the native substrates the glucosylation of TCP by the glucose-ester-forming UGT84A1 and UGT84A2 was suppressed by p-coumaric acid and sinapic acid, respectively. In contrast, the activities of UGT72E2 and UGT75D1 toward their phenolic native substrates and the xenobiotic TCP were mutually inhibited. TCP was a competitive inhibitor of sinapyl alcohol glucosylation by UGT72E2. These overlapping in vitro activities suggest cross-talk between the detoxification of xenobiotics and endogenous metabolism at the biochemical level, depending on the presence of competing substrates and enzymes.

Arabidopsis↗

Beta-tricalcium phosphate as a bone substitute for dorsal spinal fusion in adolescent idiopathic scoliosis: preliminary results of a prospective clinical study.

The aim of this study is to evaluate the ability of beta-tricalcium phosphate (TCP) in granular form to achieve dorsal spondylodesis in adolescent idiopathic scoliosis (AIS). Twenty-eight patients underwent surgical correction and were followed up for 13+/-8 (range 6-33) months. Posterolateral grafting was performed, using either autograft bone mixed with allograft bone (n=19; "bone group") or autograft bone mixed with 25 g TCP (n=9; "TCP group"). Patients were followed by clinical examination, X-rays and computed tomographic (CT) scans to measure bone mineral density. Fusion involved 12+/-1 (range 10-14) vertebrae. The segments were fused after 6+/-1 months in both groups according to the radiographs. No pseudarthrosis was observed. Bone mineral density was 430+/-111 (range 273-629) mg/cm3 in the TCP group versus 337+/-134 (range 130-669) mg/cm3 in the bone group. Resorption of TCP was complete on the radiographs after 8+/-2 (range 6-10) months. Based upon the results of this small preliminary study, the use of TCP appears to be a valuable alternative to allografts for application in the spine, even when large amounts of bone are needed.

Adolescent↗

Experimental study of bone formation around a titanium rod with beta-tricalcium phosphate and prostaglandin E2 receptor agonists.

beta-Tricalcium phosphate (beta-TCP) is an excellent bone-filling material that is completely absorbed by the body and replaced by autologous bone. Unfortunately, its mechanical strength is low, rendering its application at loaded regions difficult. The purpose of this study is to evaluate the histological and mechanical effects of single and combined use of beta-TCP and EP4 agonist on bone formation around a titanium rod. beta-TCP was loaded into the femoral bone marrow from the distal end of the femur, where the titanium implants were inserted, and the animals received twice-daily subcutaneous injections of EP4 agonist. Group I received the rod only and was designated the control group; group II received EP4 agonist only; group III received beta-TCP only; and group IV received both beta-TCP and EP4 agonist. Examination of decalcified specimens revealed favorable bone formation in all treatment groups compared with that in group I, with the most active bone formation seen in group IV. Mechanical evaluation revealed significant differences in maximum pull-out force compared with group I at weeks 4 and 8. There were no differences between groups II and III at either week 4 or 8, but the values seen in group IV at weeks 4 and 8 were significantly higher compared with the other groups. Combined use of beta-TCP and EP4 agonist is expected to compensate for bone defects resulting from revision total joint arthroplasty and to achieve stability at an early stage.

Analysis of Variance↗

Accelerated repair of a bone defect with a synthetic biodegradable bone-inducing implant.

BACKGROUND: Nothing has ever had osteoinductive capacity and degradability equivalent to that of autogenous bone, although many types of biomaterials have been developed. To address this issue, we constructed a new bone graft substitute with osteogenic potential and degradability by using porous beta-tricalcium phosphate (beta-TCP) granules, bone morphogenetic protein (BMP), and a synthetic block copolymer composed of poly-D: ,L: -lactic acid with randomly inserted p-dioxanone and polyethylene glycol (PLA-DX-PEG). In this experimental study, the bone-inducing capacity and degradation properties of the composite implant during the bone healing process were examined in vivo in a cortical and cancellous bone defect model in rabbits. METHODS: The advantages of this type of implant have been examined in a cortical bone defect model created in the distal femur of rabbits. The defects (6.5 x 5 mm) were filled with 30 mg of various implants: BMP-H [rhBMP-2, 0.0025% (w/w)], BMP-L [rhBMP-2, 0.000625% (w/w)], control A (beta-TCP alone), and control B (no implant). The distal femurs were harvested at scheduled intervals after surgery and examined for the evaluation of the bony repair of the defects by three-dimensional computed tomography and histology. RESULTS: The repair of both cortical and cancellous bone occurred predominantly in the BMP-H group, and only minor cortical bone repair and cancellous bone formation were noted in the BMP-L and control A groups. Most of the beta-TCP was resorbed in the BMP-H group at 6 weeks after surgery, whereas a significant amount of beta-TCP remained in the BMP-L and control A groups. CONCLUSIONS: beta-TCP granules coated with a BMP-retaining synthetic polymer appear to be effective in enhancing the repair of both cancellous and cortical bone defects. The early disappearance of the implanted beta-TCP and restoration of the normal anatomy of bone tissue are two notable features of this approach.

Absorbable Implants↗

Mechanism of enhanced conversion of 1,2,3-trichloropropane by mutant haloalkane dehalogenase revealed by molecular modeling.

1,2,3-Trichloropropane (TCP) is a highly toxic, recalcitrant byproduct of epichlorohydrin manufacture. Haloalkane dehalogenase (DhaA) from Rhodococcus sp. hydrolyses the carbon-halogen bond in various halogenated compounds including TCP, but with low efficiency (k (cat)/K (m )= 36 s(-1) M(-1)). A Cys176Tyr-DhaA mutant with a threefold higher catalytic efficiency for TCP dehalogenation has been previously obtained by error-prone PCR. We have used molecular simulations and quantum mechanical calculations to elucidate the molecular mechanisms involved in the improved catalysis of the mutant, and enantioselectivity of DhaA toward TCP. The Cys176Tyr mutation modifies the protein access and export routes. Substitution of the Cys residue by the bulkier Tyr narrows the upper tunnel, making the second tunnel "slot" the preferred route. TCP can adopt two major orientations in the DhaA enzyme, in one of which the halide-stabilizing residue Asn41 forms a hydrogen bond with the terminal halogen atom of the TCP molecule, while in the other it bonds with the central halogen atom. The differences in these binding patterns explain the preferential formation of the (R)- over the (S)-enantiomer of 2,3-dichloropropane-1-ol in the reaction catalyzed by the enzyme.

Hydrolases↗

Anaesthetic properties of phencyclidine (PCP) and analogues may be related to their interaction with Na+ channels.

Among other properties, phencyclidine (PCP) and analogues display anaesthetic and anticonvulsant properties. Interaction of PCP and some analogues with the voltage-sensitive Na+ channels have been investigated and compared with their interaction with the PCP receptor. PCP and TCP inhibit apparently in a competitive manner the veratridine stimulated 22Na+ synaptosomal uptake with Ki values of 8.6 and 12.7 microM, respectively, close to those obtained in the inhibition of [3H]BTX-B binding (IC50 = 4.1 and 3.8 microM, respectively). The specific [3H]TCP binding to synaptosomes in ionic near physiological conditions is inhibited by PCP and TCP with IC50 values of 1.25 and 0.29 microM, respectively. Other PCP derivatives (GK3 and GK4) and PCP-like drugs (ketamine and MK801) inhibit 22Na+ uptake in an order of potency (GK3 greater than GK4 greater than PCP greater than TCP greater than MK801 greater than ketamine) which is different from that obtained in the inhibition of [3H]TCP binding (MK801 greater than TCP greater than PCP greater than ketamine greater than GK4 greater than GK3). Ketamine inhibits the veratridine-stimulated Na+ uptake at a concentration where its anesthetic effect occurs. It was concluded that the interaction of these drugs with the Na+ channel may reflect their anaesthetic properties while the interaction with the PCP receptors may be mainly related to their anticonvulsant and ataxic properties.

Anesthetics↗

The terminal period of cancer: definition and descriptive epidemiology.

There is considerable current interest in the care of the terminally ill. This study provides information on the incidence, prevalence and duration of terminal cancer, and examines patient characteristics and patterns of care during the terminal care period (TCP). The data were derived from a retrospective study of 144 cancer deaths, utilizing information from physicians, medical records, and interviews with surviving relatives. It was estimated that 90% of all cancer deaths had a TCP, of average duration 94 days. Better functional status was associated with a longer TCP, and with spending a greater proportion of the TCP at home. Several areas of concern were noted in patterns of care, including use of a tumor-directed therapy during the TCP (in 18% of patients with a TCP) and the lack of face-to-face contact with the physician outside institutions, particularly as death approached.

Adult↗

Influence of the positioning error on 3D conformal dose distributions during fractionated radiotherapy.

The influence of patient immobilization error on 3D planned conformal radiation therapy in tumors of the thorax and pelvis was studied. The mean positioning error in 43 patients with carcinomas of the thorax and pelvis undergoing 3D conformal radiotherapy (laser supported alignment, no immobilization device) was measured. A total of 194 portal films were superposed with the corresponding simulator radiographs according to anatomic landmarks and using a subtrascope. x-, y- and z-axis deviation was determined within a coordinate system. Using specialized software including Fourier transformation the mean positioning error was employed to recalculate the dose distributions of all cases under the influence of random (Gaussian) immobilization uncertainty. The mean two-dimensional positioning error using the data from all patients was 5.5 (+/- 3.7) mm. The distribution was Gaussian. Dose volume histograms (DVHs) of each patient with and without consideration of positioning uncertainty were compared on the base of tumor control probability estimations (TCP) using published DVH reduction and TCP algorithms. Inclusion of the positioning error resulted in a mean decrease in TCP (given as the difference between the TCP assuming no positioning error and the TCP modified by the positioning error) of 2% in a series of esophagus carcinomas and of 5% in the prostate carcinomas when looking at gross tumor volume (GTV), only. Planning target volume (PTV) exhibited a relative decrease in TCP of 5% and 11%, respectively.

Algorithms↗