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Adaptation of the biobed composition for chlorpyrifos degradation to Southern Europe conditions.

Biobeds developed in Sweden bind and degrade pesticides from point sources. The objective of this work was to adapt the biobed to Italian operating conditions, for example, to identify organic materials as effective as those in the original Swedish composition. The capacity of urban and garden composts alone or mixed with citrus peel or straw to degrade chlorpyrifos and its metabolite TCP was compared to the typical Swedish biomix consisting of straw, peat, and soil. A tendency for higher 14C-chlorpyrifos mineralization and lower TCP levels was observed in the biomixes with garden compost alone or amended with straw. In a second trial, a high correlation of lower TCP with increasing levels of straw in typical Swedish biomixes was observed. Straw stimulates production of lignin-degrading enzymes such as manganese peroxidase (MnP), and further trials with pure MnP showed that this enzyme degrades TCP. Materials with an active lignin-degrading microflora are a prerequisite for effective dissipation of chlorpyrifos and non-accumulation of TCP. Thus, lignocellulosic materials as straw and garden composts should be present in biomixes to be used under Italian conditions.

Chlorpyrifos↗

Mechanical properties of sintered hydroxyapatite and tricalcium phosphate ceramic.

The ultimate values for compressive strength, Young's modulus, and toughness of cylindrical specimens of unitary aspect ratios and uniform grain-size distributions were extrapolated for hydroxyapatite (HAP) to 70 MPa, 9.2 GPa, and 0.36 J cm(-3), and for tricalcium phosphate (TCP), to 315 MPa, 21 GPa, and 2.34 J cm(-3). For total volume porosities of 50%, the corresponding values were determined: for HAP, 9.3 MPa, 1.2 GPa, 0.042 J cm(-3), for TCP, 13 MPa, 1.6 GP, 0.077 J cm(-3). Porosities of HAP specimens ranged from 3%-50%; TCP from 10%-70%. Two pore-size distributions were employed. Exponential dependencies of the mechanical properties were found upon porosity (p < 0.0001). No differences in measured mechanical properties, as determined in compression, could be attributed to pore size. The superiority of TCP increases with density and suggests that a larger or more selective pore-size distribution could be effectively employed in TCP biological implants. This work also suggests the dominant role of secondary calcium phosphates in increasing compressive strengths.

Journal Article↗

Transformation of 2,4,6-trichlorophenol by the white rot fungi Panus tigrinus and Coriolus versicolor.

The toxicity of thirteen isomers of mono-, di-, tri- and pentachlorophenols was tested in potato-dextrose agar cultures of the white rot fungi Panus tigrinus and Coriolus versicolor. 2,4,6-Trichlorophenol (2,4,6-TCP) was chosen for further study of its toxicity and transformation in liquid cultures of these fungi. Two schemes of 2,4,6-TCP addition were tested to minimize its toxic effect to fungal cultures: stepwise addition from the moment of inoculation and single addition after five days of growth. In both cases the ligninolytic enzyme systems of both fungi were found to be responsible for 2,4,6-TCP transformation. 2,6-Dichloro-1,4-hydroquinol and 2,6-dichloro-1,4-benzoquinone were found as products of primary oxidation of 2,4,6-TCP by intact fungal cultures and purified ligninolytic enzymes, Mn-peroxidases and laccases of both fungi. However, primary attack of 2,4,6-TCP in P. tigrinus culture was conducted mainly by Mn-peroxidase, while in C. versicolor it was catalyzed predominantly by laccase, suggesting a different mode of regulation of these enzymes in the two fungi.

Benzoquinones↗

Tubular cell and HIV-1 gp120 interaction products promote migration of monocytes.

Renal interstitial accumulation of monocytes is an important feature of HIV-associated nephropathy. We studied the effects of proximal tubular cell products (TCP) and proximal tubular cell-gp120 interaction products (TC-120IP) on the migration of monocytes across a modified Boyden chamber. TC-120IP promoted (P < 0.001) the migration of monocytes when compared with TCP (TCP, 45.0 +/- 5.9 vs. TC-120IP, 192.3 +/- 39.5 migrated monocytes/field). This effect of TC-120IP on monocyte migration was dose dependent. Anti-MCP-1 (TCP, 24.7 +/- 2.6; TC-120IP, 82.3 +/- 5.5; TC120-IP + anti-MCP-1 antibody, 46.5 +/- 3.5 migrated monocytes/field) as well as anti-TGF-beta antibodies (TCP, 25.8 +/- 3.4; TC120-IP, 80.3 +/- 6.9; TC-120IP + anti-TGF-beta antibody, 43.8 +/- 5.6 migrated monocytes/field) partly attenuated TC-120IP-induced migration of monocytes across a filter. Moreover, anti-MCP-1 and anti-TGF antibodies showed an additive inhibitory effect on TC-120IP-induced migration of monocytes across a filter. These results suggest that TC-120IP-induced migration of monocytes may be mediated through the generation of MCP-1 and TGF-beta by tubular cells. The present study provides the basis for a hypothesis that HIV-1 gp120 protein may be contributing to the infiltration of monocytes in the renal interstitium of patients with HIV-associated nephropathy.

AIDS-Associated Nephropathy↗

Acute effect of high-calcium milk with or without additional magnesium, or calcium phosphate on parathyroid hormone and biochemical markers of bone resorption.

OBJECTIVE: To assess whether there are any differences in the postprandial physiological responses to apple drink (control), calcium phosphate (tricalcium phosphate, TCP) and high-calcium skim milk (HCSM) with or without additional magnesium in postmenopausal women. DESIGN: Randomized, controlled, cross-over. Measurements after overnight fast before each drink, and subsequently every hour for 8 h postprandially. SETTING: Human Nutrition Studies Laboratory, Milk and Health Research Centre, Massey University, Palmerston North, New Zealand. SUBJECTS: Twenty-one healthy postmenopausal women. INTERVENTION: Four drinks, each 400 ml. (1) Apple drink (25% fruit juice). (2) TCP dispersed in water containing 1200 mg Ca. (3) HCSM containing 1200 mg Ca and 65.5 mg Mg. (4) HCSM containing 1200 mg Ca and 172 mg Mg. RESULTS: There was no difference in baseline serum calcium, PTH or C-telopeptide levels between drinks. There were no overall differences in serum calcium after apple or after either milk, but after TCP serum calcium increased from a baseline value of 2.12+/-0.08 to a mean peak of 2.21+/-0.12 mmol/l (s.d.) (P=0.0001) after 2 h. There were no significant differences in serum PTH after either apple or HCSM+Mg. In contrast, after TCP, serum PTH decreased from 2.76+/-0.69 to a mean nadir of 2.23+/-0.65 pmol/l (P=0.0001) after 1 h, and after HCSM, it decreased from 2.71+/-0.78 to a mean nadir of 2.51+/-0.87 pmol/l (P=0.007) after 2 h. Serum C-telopeptides decreased after each drink, reaching nadirs after 5 h. At this time the serum values for each of the high calcium drinks were not different from each other, but were significantly less than for apple (P=0.001 for each), being 0.22+/-0.09 ng/ml for apple, 0.15+/-0.08 for TCP, 0.14+/-0.07 for HCSM and 0.16+/-0.07 for HCSM+Mg. CONCLUSION: Despite differences in serum calcium and PTH responses to the three high-calcium drinks that we tested, there was no distinguishable difference in serum C-telopeptides between high calcium drinks.

Aged↗

Experimental study of neorectal physiology after formation of a transverse coloplasty pouch.

BACKGROUND: A novel transverse coloplasty pouch (TCP) with a larger neorectal volume than a straight coloanal anastomosis (CAA) but a smaller volume than a short colonic J pouch (CJP) may improve short-term function after rectal excision. METHODS: Twelve pigs were investigated 6 weeks after complete rectal excision followed by reconstruction with a CAA, CJP or TCP. The results were compared with findings in the normal pig rectum. RESULTS: The colonic transit times assessed by radio-opaque marker transit were 24 h for CAA, 60 h for CJP and 32 h for TCP. Non-operated control pigs had a mean transit time of 46 h. Pigs that had a CJP developed colonic dilatation and substantial faecal impaction. Colonic electrostimulation induced an adaptive relaxation in the normal rectum but a pressure increase in all neorecta, particularly after CAA. The neorectal longitudinal smooth muscle layer in pigs with a TCP was significantly thicker than that in pigs with a CAA or CJP; its thickness was closest to that of the normal pig rectum. Colonic smooth muscle layers 10 cm proximal to the coloanal anastomosis, above the neorecta, were significantly thicker after CJP than after CAA or TCP formation. No significant difference in microcirculation was observed between the three restorative procedures. CONCLUSION: Accelerated colonic transit and a lack of adequate relaxation upon endoluminal pressure increase was associated with urgency and incontinence after CAA. Delayed colonic transit, faecal impaction and ineffective muscular hypertrophy due to pouch dilatation and constipation indicated evacuation problems after CJP construction. Functional and morphometric data for TCPs suggested almost normal defaecation. Of the three restorative procedures, the data for TCPs were most similar to those obtained in the normal pig rectum at short-term follow-up.

Animals↗

Delineation of pilin domains required for bacterial association into microcolonies and intestinal colonization by Vibrio cholerae.

The toxin-co-regulated pilus (TCP), a type 4 pilus that is expressed by epidemic strains of Vibrio cholerae O1 and O139, is required for colonization of the human intestine. The TCP structure is assembled as a polymer of repeating subunits of TcpA pilin that form long fibres, which laterally associate into bundles. Previous passive immunization studies have suggested that the C-terminal region of TcpA is exposed on the surface of the pilus fibre and has a critical role in mediating the colonization functions of TCP. In the present study, we have used site-directed mutagenesis to delineate two domains within the C-terminal region that contribute to TCP structure and function. Alterations in the first domain, termed the structural domain, result in altered pilus stability or morphology. Alterations in the second domain, termed the interaction domain, affect colonization and/or infection by CTX-bacteriophage without affecting pilus morphology. In vitro and in vivo analyses of the tcpA mutants revealed that a major function of TCP is to mediate bacterial interaction through direct pilus-pilus contact required for microcolony formation and productive intestinal colonization. The importance of this function is supported by the finding that intragenic suppressor mutations that restore colonization ability to colonization-deficient mutants simultaneously restore pilus-mediated bacterial interactions. The alterations resulting from the suppressor mutations also provide insight into the molecular interactions between pilin subunits within and between pilus fibres.

Alleles↗

Degradation of 2,4,6-trichlorophenol by a specialized organism and by indigenous soil microflora: bioaugmentation and self-remediability for soil restoration.

A selected mixed culture and a strain of Alcaligenes eutrophus TCP were able to totally degrade 2,4,6,-TCP with stoichiometric release of Cl-. In cultures of Alc. eutrophus TCP, a dioxygenated dichlorinated metabolite was detected after 48 h of incubation. Experiments conducted with soil microcosms gave evidence that: the degradative process had a biotic nature and was accompanied by microbial growth; the soil used presented an intrinsic degradative capacity versus 2,4,6-TCP; the specialized organism used as inoculum was effective in degrading 2,4,6-TCP in a short time. These results could be utilized for the adoption of appropriate remediation techniques for contaminated soil.

Alcaligenes↗

Dual-setting calcium phosphate cement modified with ammonium polyacrylate.

alpha-Tricalcium phosphate bone cement, as formerly designed and developed by Driessens et al., consists of a powder composed by alpha-tricalcium phosphate (alpha-TCP) and hydroxyapatite (HA) seeds, and an aqueous solution of Na2HPO4 as mixing liquid. After mixing powder and liquid, alpha-TCP dissolves into the liquid and calcium deficient hydroxyapatite (CDHA), more insoluble than the former, precipitates as an entanglement of crystals, which causes the setting and hardening of the cement. alpha-TCP bone cement offers several advantages in comparison to calcium phosphate bioceramics and acrylic bone cements as bone graft and repairing material, like perfect adaptability to the defect size and shape, osteotransductibility, and absence of thermal effect during setting. The main handicap is its low mechanical strength. Therefore, approaching its mechanical strength to that of human bone could considerably extend its applications. In the present work, an in situ polymerization system based on acrylamide (AA) and ammonium polyacrylate (PA) as liquid reducer was added to alpha-TCP cement to increase its mechanical strength. The results showed that the addition of 20 wt% of acrylamide and 1 wt% AP to the liquid increased the compressive and tensile strength of alpha-TCP bone cement by 149 and 69% (55 and 21 MPa), respectively. The improvement in mechanical strength seems to be caused by a decrease of porosity and the reinforcing effect of a polyacrylamide network coexisting with the entanglement of CDHA crystals. The studied additives do not affect the nature of the final product of the setting reaction, CDHA, but promote the reduction of its crystal size.

Acrylic Resins↗

Genetic diversity and virulence potential of environmental Vibrio cholerae population in a cholera-endemic area.

To understand the evolutionary events and possible selection mechanisms involved in the emergence of pathogenic Vibrio cholerae, we analyzed diverse strains of V. cholerae isolated from environmental waters in Bangladesh by direct enrichment in the intestines of adult rabbits and by conventional laboratory culture. Strains isolated by conventional culture were mostly (99.2%) negative for the major virulence gene clusters encoding toxin-coregulated pilus (TCP) and cholera toxin (CT) and were nonpathogenic in animal models. In contrast, all strains selected in rabbits were competent for colonizing infant mice, and 56.8% of these strains carried genes encoding TCP alone or both TCP and CT. Ribotypes of toxigenic O1 and O139 strains from the environment were similar to pandemic strains, whereas ribotypes of non-O1 non-O139 strains and TCP(-) nontoxigenic O1 strains diverged widely from the seventh pandemic O1 and the O139 strains. Results of this study suggest that (i) the environmental V. cholerae population in a cholera-endemic area is highly heterogeneous, (ii) selection in the mammalian intestine can cause enrichment of environmental strains with virulence potential, (iii) pathogenicity of V. cholerae involves more virulence genes than currently appreciated, and (iv) most environmental V. cholerae strains are unlikely to attain a pandemic potential by acquisition of TCP and CT genes alone. Because most of the recorded cholera pandemics originated in the Ganges Delta region, this ecological setting presumably favors extensive genetic exchange among V. cholerae strains and thus promotes the rare, multiple-gene transfer events needed to assemble the critical combination of genes required for pandemic spread.

Animals↗

A monooxygenase catalyzes sequential dechlorinations of 2,4,6-trichlorophenol by oxidative and hydrolytic reactions.

Ralstonia eutropha JMP134 2,4,6-trichlorophenol (2,4,6-TCP) 4-monooxygenase catalyzes sequential dechlorinations of 2,4,6-TCP to 6-chlorohydroxyquinol. Although 2,6-dichlorohydroxyquinol is a logical metabolic intermediate, the enzyme hardly uses it as a substrate, implying it may not be a true intermediate. Evidence is provided to support the proposition that the monooxygenase oxidized 2,4,6-TCP to 2,6-dichloroquinone that remained with the enzyme and got hydrolyzed to 2-chlorohydroxyquinone, which was chemically reduced by ascorbate and NADH to 6-chlorohydroxyquinol. When the monooxygenase oxidized 2,6-dichlorophenol, the product was 2,6-dichloroquinol, which was not further converted to 6-chlorohydroxyquinol, implying that the enzyme only converts 2,6-dichloroquinone to 6-chlorohydroxyquinol. Stoichiometric analysis indicated the consumption of one O2 molecule per 2,4,6-TCP converted to 6-chlorohydroxyquinol, ruling out the possibility of two oxidative reactions. Experiments with 18O-labeling gave direct evidence for the incorporation of oxygen from both O2 and H2O into the produced 6-chlorohydroxyquinol. A monooxygenase that catalyzes hydroxylation by both oxidative and hydrolytic reactions has not been reported to date. The ability of the enzyme to perform two types of reactions is not due to the presence of a second functional domain but rather is due to catalytic promiscuity, as a homologous monooxygenase converts 2,4,6-TCP to only 2,6-dichloroquinol. Employing both conventional catalysis and catalytic promiscuity of a single enzyme in two consecutive steps of a metabolic pathway has been unknown previously.

Catalysis↗

Microbial dechlorination of 2,4,6-trichlorophenol in anaerobic sewage sludge.

The dechlorination of 2,4,6-trichlorophenol (TCP) in municipal sewage sludge with a chlorophenol (CP)-adapted consortium was investigated. Results show that dechlorination rates differed according to the source of the sludge samples used in the batch experiments. No significant differences in 2,4,6-TCP dechlorination were observed following treatment with inoculum at densities ranging from 10% to 50% (V/V), but a significant delay was noted at 5% (V/V) density. Overall, results show that the higher the 2,4,6-TCP concentration, the slower the dechlorination rate. The addition of acetate, lactate, pyruvate, vitamin B12 or manganese dioxide did not results in a significant change in 2,4,6-TCP dechlorination. Data collected from a bioreactor experiment revealed that pH 7.0 and a total solid concentration of 10 g/L were optimal for dechlorination. Dechlorination rates decreased significantly at higher agitation speeds. 2,4,6-TCP dechlorination was enhanced under methanogenic conditions, but it was inhibited under denitrifying and sulfate-reducing conditions.

Anaerobiosis↗

Natural vitamin E alpha-tocotrienol: retention in vital organs in response to long-term oral supplementation and withdrawal.

The natural vitamin E tocotrienol (TCT) possesses biological properties not shared by tocopherols (TCP). Nanomolar alpha-TCT, not alpha-TCP, is potently neuroprotective (JBC 275:13049; 278:43508; Stroke 36:2258). The report that the affinity of TTP to bind (alpha-TCT is an order of magnitude lower than that for alpha-TCP questions the bioavailability of orally taken TCT to tissues. Oral supplementation of TCT for 3 years in nine generations of female and male rat was studied. Ten vital organs were examined. To gain insight into the turnover of alpha-TCT in tissues, a subset of supplemented rats was moved to vitamin E deficient diet for 7 weeks. Orally supplemented alpha-TCT was delivered to all vital organs including the brain and spinal cord in significant amounts. In organs such as the skin, adipose and gonads the maximum level of alpha-TCT achieved in response to supplementation was folds higher than baseline values of alpha-TCP in rats maintained on laboratory chow. Females had higher levels of alpha-TCT compared to matched tissues of corresponding males. To gain insight into how quickly alpha-TCT is metabolized in the tissues, washout of alpha-TCT from vital organs was examined. alpha-TCT accumulated in vital organs over more than 2 years was almost completely lost in less than 2 months when the supplementation was stopped. This is in sharp contrast with findings related to alpha-TCP retention. The ability of long-term oral supplementation to maintain and elevate alpha-TCT levels in vital organs together with the rapid elimination of the intact vitamin from all organs studied underscores the need for continuous oral supplementation of TCT.

Adipose Tissue↗

Some characteristics of tumour control probability for heterogeneous tumours.

Some mathematical characteristics of tumour control probability (TCP) have been examined in the light of potential intra-tumour variations in clonogenic cell characteristics. In particular, an explicit expression for the relationship between the dose distribution and the probability of local clonogen eradication is obtained which maximizes the TCP using both general and specific TCP models, for the case of fixed energy deposition into the tumour volume. Characteristics of this expression are considered for the cases of varying clonogen cell density (uniform radiosensitivity) and uniform cell density (uniform radiosensitivity), yielding the 'uniform TCP' and 'uniform dose' results respectively for TCP maximization. These situations are examined graphically to highlight the possible consequences of neglecting intra-tumour heterogeneity in dose prescription.

Cell Survival↗

Tumour control probability: a formulation applicable to any temporal protocol of dose delivery.

An analytic expression for the tumour control probability (TCP), valid for any temporal distribution of dose, is discussed. The TCP model, derived using the theory of birth-and-death stochastic processes, generalizes several results previously obtained. The TCP equation is [equation: see text] where S(t) is the survival probability at time t of the n clonogenic tumour cells initially present (at t = 0), and b and d are, respectively, the birth and death rates of these cells. Equivalently, b = 0.693/Tpot and d/b is the cell loss factor of the tumour. In this expression t refers to any time during or after the treatment; typically, one would take for t the end of the treatment period or the expected remaining life span of the patient. This model, which provides a comprehensive framework for predicting TCP, can be used predictively, or--when clinical data are available for one particular treatment modality (e.g. fractionated radiotherapy)--to obtain TCP-equivalent regimens for other modalities (e.g. low dose-rate treatments).

Brachytherapy↗

Adapting radiotherapy to hypoxic tumours.

In the current work, the concepts of biologically adapted radiotherapy of hypoxic tumours in a framework encompassing functional tumour imaging, tumour control predictions, inverse treatment planning and intensity modulated radiotherapy (IMRT) were presented. Dynamic contrast enhanced magnetic resonance imaging (DCEMRI) of a spontaneous sarcoma in the nasal region of a dog was employed. The tracer concentration in the tumour was assumed related to the oxygen tension and compared to Eppendorf histograph measurements. Based on the pO(2)-related images derived from the MR analysis, the tumour was divided into four compartments by a segmentation procedure. DICOM structure sets for IMRT planning could be derived thereof. In order to display the possible advantages of non-uniform tumour doses, dose redistribution among the four tumour compartments was introduced. The dose redistribution was constrained by keeping the average dose to the tumour equal to a conventional target dose. The compartmental doses yielding optimum tumour control probability (TCP) were used as input in an inverse planning system, where the planning basis was the pO(2)-related tumour images from the MR analysis. Uniform (conventional) and non-uniform IMRT plans were scored both physically and biologically. The consequences of random and systematic errors in the compartmental images were evaluated. The normalized frequency distributions of the tracer concentration and the pO(2) Eppendorf measurements were not significantly different. 28% of the tumour had, according to the MR analysis, pO(2) values of less than 5 mm Hg. The optimum TCP following a non-uniform dose prescription was about four times higher than that following a uniform dose prescription. The non-uniform IMRT dose distribution resulting from the inverse planning gave a three times higher TCP than that of the uniform distribution. The TCP and the dose-based plan quality depended on IMRT parameters defined in the inverse planning procedure (fields and step-and-shoot intensity levels). Simulated random and systematic errors in the pO(2)-related images reduced the TCP for the non-uniform dose prescription. In conclusion, improved tumour control of hypoxic tumours by dose redistribution may be expected following hypoxia imaging, tumour control predictions, inverse treatment planning and IMRT.

Animals↗

Optimum parameters in a model for tumour control probability, including interpatient heterogeneity: evaluation of the log-normal distribution.

The heterogeneity of human tumour radiation response is well known. Researchers have used the normal distribution to describe interpatient tumour radiosensitivity. However, many natural phenomena show a log-normal distribution. Log-normal distributions are common when mean values are low, variances are large and values cannot be negative. These conditions apply to radiosensitivity. The aim of this work was to evaluate the log-normal distribution to predict clinical tumour control probability (TCP) data and to compare the results with the homogeneous (delta-function with single alpha-value) and normal distributions. The clinically derived TCP data for four tumour types-melanoma, breast, squamous cell carcinoma and nodes-were used to fit the TCP models. Three forms of interpatient tumour radiosensitivity were considered: the log-normal, normal and delta-function. The free parameters in the models were the radiosensitivity mean, standard deviation and clonogenic cell density. The evaluation metric was the deviance of the maximum likelihood estimation of the fit of the TCP calculated using the predicted parameters to the clinical data. We conclude that (1) the log-normal and normal distributions of interpatient tumour radiosensitivity heterogeneity more closely describe clinical TCP data than a single radiosensitivity value and (2) the log-normal distribution has some theoretical and practical advantages over the normal distribution. Further work is needed to test these models on higher quality clinical outcome datasets.

Breast Neoplasms↗

Use of a biphasic graft constructed with chondrocytes overlying a beta-tricalcium phosphate block in the treatment of rabbit osteochondral defects.

To evaluate the ability of a biphasic construct to repair osteochondral defects in articular cartilage, plugs made of chondrocytes in collagen gel overlying a resorbable porous beta-tricalcium phosphate (TCP) block were implanted into defects in rabbit knees. The repair tissue was evaluated at 8, 12, and 30 weeks. Eight weeks after implantation of the biphasic construct, histologic examination showed hyaline-like cartilage formation that was positive for safranin O and type II collagen. At 12 weeks, most of the beta-TCP was replaced by bone, with a small amount remaining in the underlying cartilage. In the cell-seeded layer, the newly formed middle and deep cartilage adjacent to the subchondral bone stained with safranin O, but no staining was observed in the superficial layer. In addition, cell morphology was distinctly different from the deep levels of the reparative cartilage, with hypertrophic cells at the bottom of the cartilaginous layer. At 30 weeks, beta-TCP had completely resorbed and a tidemark was observed in some areas. In contrast, controls (defects filled with a beta-TCP block alone) showed no cartilage formation but instead had subchondral bone formation. These findings indicate that beta-TCP-supported chondrocytes in collagen gel can partially repair isolated articular cartilage osteochondral defects.

Animals↗