RENEB Inter-Laboratory Comparison 2021: The FISH-Based Translocation Assay.

Translocation analysis using fluorescence in situ hybridization (FISH) is the method of choice for dose assessment in case of chronic or past exposures to ionizing radiation. Although it is a widespread technique, unlike dicentrics, the number of FISH-based inter-laboratory comparisons is small. For this reason, although the current Running the European Network of Biological and Physical retrospective Dosimetry (RENEB) inter-laboratory comparison 2021 was designed as a fast response to a real emergency scenario, it was considered a good opportunity to perform an inter-laboratory comparison using the FISH technique to gain further experience. The Bundeswehr Institute of Radiobiology provided peripheral blood samples from one healthy human volunteer. Three test samples were irradiated with blinded doses of 0, 1.2, and 3.5 Gy, respectively. Samples were then sent to the seven participating laboratories. The FISH technique was applied according to the standard procedure of each laboratory. Both, the frequency of translocations and the estimated dose for each sample were sent to the coordinator using a special scoring sheet for FISH. All participants sent their results in due time. However, although it was initially requested to send the results based on the full analysis, evaluating 500 equivalent cells, most laboratories only sent the results based on triage, with a smaller number of analyzed cells. In the triage analysis, there was great heterogeneity in the number of equivalent cells scored. On the contrary, for the full analysis, this number was more homogeneous. For all three samples, one laboratory showed outlier yields compared to the other laboratories. Excluding these results, in the triage analysis, the frequency of translocations in sample no. 1 ranged from 0 to 0.013 translocations per cell, and for samples no. 2 and no. 3 the genomic mean frequency were 0.27 ± 0.03 and 1.47 ± 0.14, with a coefficient of variation of 0.29 and 0.23 respectively. Considering only results obtained in the triage analysis for sample no. 1, all laboratories, except one, classified this sample as the non-irradiated one. For sample no. 2, excluding the outlier value, the mean reported dose was 1.74 ± 0.16 Gy indicating a mean deviation of about 0.5 Gy to the delivered dose of 1.2 Gy. For sample no. 3 the mean dose estimated was 4.21 ± 0.21 Gy indicating a mean deviation of about 0.7 Gy to the delivered dose of 3.5 Gy. In the frame of RENEB, this is the second FISH-based inter-laboratory comparison. The whole exercise was planned as a response to an emergency, therefore, a triage analysis was requested for all the biomarkers except for FISH. Although a full analysis was initially requested for FISH, most of the laboratories reported only a triage-based result. The main reason is that it was not clearly stated what was required before starting the exercise. Results show that most of the laboratories successfully discriminated unexposed and irradiated samples from each other without any overlap. A good agreement in the observed frequencies of translocations was observed but there was a tendency to overestimate the delivered doses. Efforts to improve the harmonization of this technique and subsequent exercises to elucidate the reason for this trend should be promoted.

A rare case of retroperitoneal primitive neuroectodermal tumor (PNET).

Retroperitoneal primitive neuroectodermal tumors (PNET) is a rare neoplastic disease of high malignancy with a tendency towards early metastasis, affect young adults irrespective of the gender. We present 81 year old woman, who was admitted in the Urology Department with symptoms of right flank pain and hematuria. Contrast-enhanced computerized tomography scan (CT-scan) showed a large heterogeneous right kidney mass around 12 cm in diameter. The final diagnosis of primitive neuroectodermal tumor (PNET) was established based on clinical, pathological, and molecular results.

Management of Leydig cell tumors of the testis-a case report.

We report 1 case of Benign Leydig cell tumor. A 45-year-old male was admitted to the Urology department with a large painless mass in the right testis of 1 year duration. The patient underwent radical high right orchiectomy, with a preliminary diagnosis of right testicular tumor. On the basis of the pathologic and immunohistochemical findings, the testicular mass was diagnosed as a benign Leydig cell tumor. Long-term follow-up is necessary to exclude recurrence or metastasis and also the endocrine profile and imaging investigations need to be repeated periodically.

Model-predicted occurrence of multiple pharmaceuticals in Swedish surface waters and their flushing to the Baltic Sea.

An exposure assessment for multiple pharmaceuticals in Swedish surface waters was made using the STREAM-EU model. Results indicate that Metformin (27 ton/y), Paracetamol (6.9 ton/y) and Ibuprofen (2.33 ton/y) were the drugs with higher amounts reaching the Baltic Sea in 2011. 35 of the studied substances had more than 1 kg/y of predicted flush to the sea. Exposure potential given by the ratio amount of the drug exported to the sea/amount emitted to the environment was higher than 50% for 7 drugs (Piperacillin, Lorazepam, Metformin, Hydroxycarbamide, Hydrochlorothiazide, Furosemide and Cetirizine), implying that a high proportion of them will reach the sea, and below 10% for 27 drugs, implying high catchment attenuation. Exposure potentials were found to be dependent of persistency and hydrophobicity of the drugs. Chemicals with Log D > 2 had exposure potentials <10% regardless of their persistence. Chemicals with Log D  <  -2 had exposure potentials >35% with higher ratios typically achieved for longer half-lives. For Stockholm urban area, 17 of the 54 pharmaceuticals studied had calculated concentrations higher than 10 ng/L. Model agreement with monitored values had an r2 = 0.62 for predicted concentrations and an r2 = 0.95 for predicted disposed amounts to sea.

Evaluation of human pharmaceutical emissions and concentrations in Swedish river basins.

An emissions inventory for top consumed human pharmaceuticals in Sweden was done based on national consumption data, human metabolic rates and wastewater treatment removal rates. Concentrations of pharmaceuticals in surface waters in Swedish river basins were predicted using estimated emissions from the inventory and river discharges. Our findings indicate that the top ten emitted pharmaceuticals in our study set of 54 substances are all emitted in amounts above 0.5ton/y to both surface waters and soils. The highest emissions to water were in decreasing order for Metformin, Furosemide, Gabapentin, Atenolol and Tramadol. Predicted emissions to soils calculated with the knowledge that in Sweden sludge is mostly disposed to soil, point to the highest emissions among the studied drugs coming from, in decreasing order, Metformin, Paracetamol, Ibuprofen, Gabapentin and Atenolol. Surface water concentrations in Sweden's largest rivers, all located in low density population zones, were found to be below 10ng/L for all substances studied. In contrast, concentrations in surface waters in Stockholm's metropolitan area, the most populous in Sweden, surpassed 100ng/L for four substances: Atenolol, Metformin, Furosemide and Gabapentin.

A novel thermostable neutral proteinase from Saccharomonospora canescens.

A novel thermostable neutral proteinase, called NPS, was purified to electrophoretic homogeneity from the culture broth of Saccharomonospora canescens sp. novus, strain 5. The molecular mass was determined by SDS-polyacrylamide gel electrophoresis to be 35,000 Da. The enzyme exhibits a sharp pH optimum of proteolytic activity at pH 6.7. NPS was completely inactivated with inhibitors, typical for metalloendopeptidases, EDTA and 1,10-phenantroline, whereas the serine proteinase inhibitor PMSF had no effect. Atomic absorption measurements showed that the proteinase binds a single zinc and four calcium ions. The enzyme thermostability was characterized in the absence and presence of added calcium. Melting temperature, Tm = 77 degrees C and an activation energy, Ea, for the thermal deactivation of the excited protein fluorophores of 72.13 kJ mol-1 were calculated in the presence of 100 mM CaCl2. The Ea-value is considerably higher than those obtained for a number of proteinases from microorganisms and was explained by the thermostable structure of the enzyme. Effective radiationless energy transfer from phenol groups to indole rings was observed. 68% of the light absorbed by tyrosyl residues is transferred to tryptophyl side chains. No homology was found after comparison of the NPS N-terminal sequence, including the first 26 residues, with those of other neutral proteinases from microorganisms. In contrast to the well-known bacterial neutral proteinase thermolysin and related enzymes from microorganisms, NPS possesses arylamidase and esterase activities. Further crystallographic studies will reveal the structural reasons for this specificity. Epoxy and epithio pyranosides are inhibitors of the proteinase arylamidase activity.

A novel thermostable inhibitor of trypsin and subtilisin from the seeds of Brassica nigra: amino acid sequence, inhibitory and spectroscopic properties and thermostability.

A novel thermostable protein inhibitor of trypsin and subtilisin, called BN, was isolated from the seeds of Brassica nigra. The purified protein gave a single band on SDS-PAGE, corresponding to a molecular mass of 15 500 +/- 1000 Da. The inhibitor is composed of two disulfide-linked polypeptide chains, consisting of 39 and 90 residues, respectively. The amino acid sequence of the two chains was determined by Edman degradation of peptides, isolated from enzyme hydrolysates with TPCK-trypsin, EndoLysC proteinase and a Glu-specific proteinase of reduced and vinylpyridinated protein samples. A segment of the 'heavy' chain, between residues 65 and 81, showed homology with the reactive site loop region of the 6-kDa trypsin inhibitors from Nicotiana alata. The basic residue in position 39 (N. alata) or 70 (napins) is conserved as arginine or lysine in all inhibitors from N. alata and in all napins hitherto sequenced. Probably, the two families of trypsin inhibitors have structurally similar reactive sites. BN exhibits an extremely high thermostability: CD measurements showed that during heating to 97 degrees C it preserves a considerable part of the polypeptide backbone folding. Studies on the fluorescence properties of the inhibitor BN in the absence and presence of neutral or ionic quenchers demonstrated that the intrinsic emission of this protein is dominated by a tryptophyl residue, buried in the interior of the protein matrix. 20% of the light absorbed by Tyr 63 of the 'heavy' chain is transferred to Trp 26 of the 'light' chain.

The role of angiotensin II and of its receptor subtypes in the acetic acid-induced abdominal constriction test.

The effects of angiotensin II (AngII), the AngII analogues saralasin--[Sar1, Ala8]AngII, sarmesin--[Sar1Tyr(Me)4]AngII, the nonpeptide AngII receptor antagonists DuP753 (losartan) (for AT1 receptor subtype) and PD123319 (for AT2 receptor subtype), as well as combinations of AngII and each of its analogues and receptor antagonists, administered intracerebroventricularly (ICV), were studied on mice using the acetic acid-induced abdominal constrictions test (acetic acid 1% intraperitoneally, IP). The abdominal constrictions were counted at 5-min intervals for 30 min. AngII at doses of 0.05, 0.1, and 1 microg exerted a dose-dependent antinociceptive effect. Saralasin, sarmesin, losartan, and PD123319 exhibited a dose-dependent effect on nociception: they either increased or decreased it. PD123319 antagonized the antinociceptive effect of AnglI while losartan was ineffective. The importance of AT2 receptor subtype for the nociception reducing effect of AngII is considered.

Characterisation of Trichinella isolates from Bulgaria by molecular typing and cross-breeding.

Trichinella spp. larvae were collected from domestic and wild-life animals in association with 15 human trichinellosis outbreaks registered between 1999-2002 in Bulgaria. Furthermore, Trichinella spp. isolates were obtained from 62 naturally infected wild animals and of a rat. All isolates were subjected to speciation by both multiplex PCR and cross-breeding experiments. Epidemiological and clinical data were collected and analysed using standard protocols for epidemiological surveillance and control of outbreaks. Only two species were identified-Trichinella britovi and Trichinella spiralis. Results obtained by molecular typing fully matched those of cross-breeding. More specifically, parasite isolates obtained upon 15 epidemic outbreaks revealed the predominance of T. britovi (n = 10) when compared to T. spiralis (n = 5). With regard to host origin, the predominant species detected among wild boar was T. britovi (n = 4), and T. spiralis was identified in one wild boar sample only. Among the isolates obtained from domestic pig products, T. britovi was found in five cases and T. spiralis in four cases, respectively. In the naturally infected wild animals not related to epidemics, only T. britovi was demonstrated. The present results provide a strong indication that both T. britovi and T. spiralis operate within domestic and sylvatic cycles in Bulgaria. Geographically, the distribution of T. britovi appears to include Central, Southern, Eastern and Western parts of the country, and wildlife animals from the Mid Balkan Mountains and Mid Sredna Gora Mountains, T. spiralis was found in Western and Southwestern Bulgaria, only.

Steady-state and time-resolved fluorescence of Esperase: comparison with the X-ray structure in the region of the two tryptophans.

Fluorescence emission properties of the alkaline protease Esperase have been investigated using steady-state and time-resolved fluorescence spectroscopy. The local polarity and solvent accessibility of the tryptophyl chromophores is characterized. Quenching studies demonstrated that Trp 6 and Trp 113 are 'buried' to acrylamide, iodide ions and caesium ions. An abnormally low tryptophan quantum yield was calculated showing that the emission of the two indole rings is significantly quenched by nearby side chains or peptide bonds. The fluorescence decay of PMS-Esperase was well fitted by two exponentials with lifetimes of 2.7 and 0.35 ns. X-ray data for Esperase (S. Klupsch, Ph.D. Thesis, University of Hamburg, Hamburg, Germany) in the region of the two tryptophans were used to explain the observed emission properties. Gln 182 and Asn 204 as well as Asn 117 and Met 119 are the most likely quenchers, respectively, of the Trp 6 and Trp 113 fluorescence. The two tryptophans in Esperase are 'buried' in hydrophobic regions and are excellent intrinsic probes to study folding-unfolding reactions. Experiments in the presence and absence of added calcium ions demonstrated the stabilizing role of the Ca(2+)-binding sites.

Spectroscopic investigation of calcium binding sites in the neurotoxin vipoxin and its components-relation with the X-ray structure.

Vipoxin is a neurotoxin from the venom of Vipera ammodytes meridionalis, the most toxic snake in Europe. It is a unique complex of a toxic phospholipase A2 (PLA2) and a non-toxic PLA2-like protein inhibitor (Inh) which probably evolved from the enzyme and reduces its activity and toxicity. The enzymatic activity of Vipoxin is Ca2+-dependent and the interaction of this metal ion with the neurotoxic complex and its separated components was investigated using the fluorescent probe ANS. Vipoxin binds two calcium ions, one per each subunit. The X-ray model of the Ca2+-free neurotoxin shows that the potential metal-binding sites require minor structural changes to bind calcium. The dissociation constants K(2+)Ca of the calcium complexes of Vipoxin and its components, PLA2 and Inh, were determined to be 16, 10 and 9 mM, respectively. The affinity for calcium of Vipoxin is reduced in comparison to those of PLA2 and Inh. The X-ray model shows that the potential Ca2+-binding sites in the two components are partially 'shielded' in the complex. The affinity of the neurotoxin to Sr2+ and Ba2+ is lower and the respective K(2+)Ca are 20 and 30 mM. The saturation of Ca2+-binding sites increased the melting point Tm of Vipoxin by 11 degrees C and the activation energy for the thermal deactivation of the excited tryptophans Ea by 11 kJ mol(-1) x Ca2+ is important not only for the enzymatic activity of Vipoxin but also for its thermostability.

Spectroscopic investigation of phenolic groups ionization in the vipoxin neurotoxic phospholipase A2: comparison with the X-ray structure in the region of the tyrosyl residues.

The neurotoxin vipoxin is the major lethal component of the venom of Vipera ammodites meridionalis, the most toxic snake in Europe. It is a complex between a toxic phospholipase A2 (PLA2) and a non-toxic protein inhibitor (Inh). Tyrosyl residues are involved in the catalytic site (Tyr 52 and 73) and in the substrate binding (Tyr 22). Spectroscopic studies demonstrated differences in the ionization behavior of the various phenolic hydroxyl groups in the toxic PLA2. The tyrosyl side chains of the enzyme can be classified into three groups: (a) three phenolic hydroxyls are accessible to the solvent and titrate normally, with a pKeff = 10.45; (b) three residues are partially 'buried' and participate in hydrogen bonds with neighboring functional groups. They titrate anomalously with a pKeff = 12.17; (c) two tyrosines with a pKeff = 13.23 are deeply 'buried' in the hydrophobic interior of PLA2. They became accessible to the titrating agent only after alkaline denaturation of the protein molecule. The spectroscopic data are related to the X-ray structure of the vipoxin PLA2. The refined model was investigated in the region of the tyrosyl side chains. The accessible surface area of each tyrosyl residue and each phenolic hydroxyl group was calculated. A good correlation between the spectrophotometric and the crystallographic data was observed. The ionization behavior of the phenolic groups is explained by peculiarities of the protein three-dimensional structure and the participation of tyrosines in the catalytic site hydrogen bond network. Attempts are made to assign the calculated pKeff values to individual residues. The high degree of 'exposure' on the protein surface of Tyr 22 and 75 is probably important for their function as parts of the substrate binding and pharmacological sites.

Penaeus monodon (tiger shrimp) hemocyanin: subunit composition and thermostability.

Penaeus monodon (class Crustacea, order Decapoda) is one of the largest shrimps of the Penaeidea family from the Indo-West Pacific region. The dioxygen-transporting protein hemocyanin, isolated from the hemolymph of this invertebrate, is composed of three 75-76 kDa structural/functional subunits designated as Pm1, Pm2 and Pm3. The N-terminal sequences of the chains were determined and compared with those of other decapodan hemocyanin subunits. Pm2 and Pm3 are highly homologous and electrophoretically undistinguishable polypeptides. In comparison to Pml, they have an extension of six residues. Pm1 is closely related to the subunit Pv2 of the Penaeus vannamei hemocyanin. Probably, subunits like Pm1 and Pv2 are family-specific for the Penaeidea hemocyanins and the other subunits are species-specific. Comparison of N-terminal sequences of respiratory proteins from the sub-orders Natantia and Reptantia demonstrated family- and sub-order-specific sequences. A melting point of 69 degrees C, lower than those for the di-hexameric decapodan hemocyanins, was determined from the temperature dependence of ellipticity of the mono-hexameric Penaeus monodon hemocyanin. Thermostability of decapodan hemocyanins depends on their aggregation state.

Viviparus ater hemocyanin: investigation of the dioxygen-binding site and stability of the oxy- and apo-forms.

The active site of Viviparus ater (mollusc) hemocyanin was investigated using the fact that the binding of dioxygen to the binuclear copper-containing sites of hemocyanins is connected with the appearance of specific dichroic bands which are very sensitive to changes in the structrure and polarity of the environment. Oxy-Viviparus ater hemocyanin exhibits near UV and visible circular dichroism spectra different from those of other molluscan and arthropodan hemocyanins. These differences are due probably to variations in the geometry or charge distribution in the dioxygen binding sites of the compared proteins. The thermostability of Viviparus ater hemocyanin and the significance of the copper-dioxygen system for the stability were also investigated. "Melting" temperatures, Tm, of 77 degrees C for the oxy-hemocyanin and 57 degrees C for the apo-protein were calculated from the denaturation curves which demonstrates the considerable role of the binuclear active site for the thermostability. Viviparus ater hemocyanin is more thermostable than other hemocyanins for which data are published.

Significance of ultrasound in the diagnosis and treatment of achilles tendon rupture.

UNLABELLED: The aim of this study is to show the importance of ultrasound method in the diagnosis of Achilles tendon rupture, the choice of method of treatment and monitoring of treatment using the same method. MATERIAL AND METHODS: Between 1999 and 2009, 134 patients with Achilles tendon rupture were referred to our Clinic. 66 patients (with a mean age of 38 years) were treated with surgical suture followed by plaster immobilization for a period of 8 weeks. 68 patients (with a mean age of 42 years) were treated conservatively with plaster immobilization for a period of 8 weeks. The follow-up in both groups of patients was 2 years. RESULTS: During the clinical and ultrasound monitoring of the patients it was proved that repeated rupture of the same tendon occurs on average within 12 months. Return to sports activities showed in 57% of the conservatively treated patients and in 55% of surgically treated patients. The patients with Achilles tendon rupture were treated at our Clinic with previously standardized protocol which, besides the clinical examination, used the ultrasound method. SUMMARY: Ultrasound examination is a very important method in the diagnosis and the choice of the method of treatment, as well as in the evaluation of results in patients with Achilles tendon rupture, either in operative or conservative treatment.

Prevention, management and rehabilitation of patients with cerebral palsy.

UNLABELLED: The aim of this paper is to present the importance of orthopaedics in the prevention, treatment and rehabilitation of persons with cerebral palsy. MATERIAL AND METHODS: The paper is based on a study realized at the University Orthopaedic Clinic, Medical School, Ss. Cyril and Methodius University in Skopje, with application of documentary analysis, observation and clinical analysis of 76 patients with cerebral palsy, aged between 1 and 15 years. RESULTS: Orthopaedics is a surgical field of medicine, in which context there have been, are and will be, as a challenge, many questions for the present and future generations. Data analysis pointed out that surgical treatment in combination with conservative treatment give a hope that a definite or prolonged cure is possible, with maximal and possibly preserved function of the loco-motor system or, in the most serious cases, abatement of the impaired locomotor system symptoms. SUMMARY: Early diagnosis and appropriate application of surgical treatment enable better functioning of persons with cerebral palsy by maximal exploitation of their remaining abilities.

Simulation of chemical metabolism for fate and hazard assessment. III. New developments of the bioconcentration factor base-line model.

The new development of the bioconcentration factor (BCF) base-line model of Dimitrov et al. [SAR QSAR Environ. Res. 6 (2005), pp. 531-554] is presented. The model applicability domain was expanded by enlarging the training set of the model up to 705 chemicals. The list of chemical-dependent mitigating factors was expanded by including water solubility of chemicals. The original empirical term for estimating ionization of chemicals was mechanistically analysed using two different approaches. In the first one, the ionization potential of chemicals was estimated based on the acid dissociation constant (pK(a) ). This term was found to be less adequate for inclusion in the ultimate BCF model, due to overestimating ionization of chemicals. The second approach, estimating the ionization as a ratio between distribution and partition coefficients (log P and log D), was found to be more successful. The new ionization term allows modelling of chemicals with both acidic and basic functionalities and chemicals undergoing different degrees of ionization. The significance of the different mitigating factors which can reduce the maximum bioconcentration potential of the chemicals was re-formulated and model parameters re-evaluated.

Clinical and radiographic analysis of the operative procedure results according to the method of Mitchell and Keller used for correction of hallux valgus deformities.

UNLABELLED: The aim of this study is to make a correlation of the clinical and radiographic results after performing two different surgical procedures for correction of hallux valgus deformity. MATERIAL AND METHODS: The study included 70 patients having hallux valgus deformity of the foot, and they were divided into two groups. The first group (Group 1) was composed of 35 patients who were treated by osteotomy of the I-st metatarsal bone according to Mitchell, while the second group (Group 2) was also composed of 35 patients who were treated by resectional arthroplasty according to Keller. Clinical (pain and metatarsalgia, as well as most dominant symptoms) and radiographic examinations (I metatarsophalangeal angle and I intermetatarsal angle) were analysed comparatively during the evaluation. The analysis of the clinical and radiographic results was performed pre-operatively and post-operatively for the two groups. RESULTS: According to their sex, the patients were 5 men and 65 women. Using the method of Mitchell, pain as a clinical symptom post-operatively was found in only 3 patients out of the 35 with operated feet, while in the other group of patients treated by the method of Keller, there was no presence of pain in any of the patients. Comparatively, this does not present a statistically significant difference (p>0.05). Nor do, the differences in the distribution of metatrsalgia incidence show a statistical significance between the two groups (p>0.05). There is no significant difference (p>0.05) in the patients of the two groups concerning the pre-operative mean dimension values of the I metatarsophalangeal angle and I intermetatarsal angle. However, the radiographic analysis of the same angles in both groups, one year post-operatively, showed a high statistically significant difference (p<0.001). SUMMARY: Mitchell's operative technique could be recommended as an effective procedure for the correction of hallux valgus and metatarsus primus varus in young and middle-aged patients, while the resection arthroplasty according to the method of Keller is recommended for older patients with arthrotic changes.

The role of clinical diagnosis in meniscal lesions of the knee.

The aim of the present paper is to evaluate the importance of clinical examination in the process of diagnosing meniscal lesions, as well as to establish the accuracy of clinical examination in comparison to the arthroscopic diagnosis. In the last 5 years, in the Orthopedic Surgery Clinic in Skopje, the authors have diagnosed and treated 205 patients suffering from meniscal lesions. The method of anamnestic testing, and of clinical examination and the statistical method have been applied. All the patients were subjected to the following tests: well taken anamnesis of the knee injury mechanism, standard orthopedic examination (Stainman I and II, McMurray and Appley test), standard radiographic images of the knee, additional examination with MRI for some of the patients and arthroscopic examination. The results obtained with clinically established diagnosis were compared to the results obtained with arthroscopic diagnosis. Comparing the results of the clinical and arthroscopic diagnoses, an insufficiency of the clinical diagnosis of almost 20% was established. The results are close to those given in the professional literature which refers to the standardized approach in taking the anamnestic data and the realization of the physical signs and tests examination technique. It means that the physical signs and tests, as well as the well-taken anamnesis, are the foundation for establishing a diagnosis of meniscal lesions. Arthroscopy is truly the only golden standard for the diagnosis of all entities of the knee joint, including meniscal lesions.

Simulation of chemical metabolism for fate and hazard assessment. II CATALOGIC simulation of abiotic and microbial degradation.

The unprecedented pollution of the environment by xenobiotic compounds has provoked the need to understand the biodegradation potential of chemicals. Mechanistic understanding of microbial degradation is a premise for adequate modelling of the environmental fate of chemicals. The aim of the present paper is to describe abiotic and biotic models implemented in CATALOGIC software. A brief overview of the specificities of abiotic and microbial degradation is provided followed by detailed descriptions of models built in our laboratory during the last decade. These are principally new models based on unique mathematical formalism already described in the first paper of this series, which accounts more adequately than currently available approaches the multipathway metabolic logic in prokaryotes. Based on simulated pathways of degradation, the models are able to predict quantities of transformation products, biological oxygen demand (BOD), carbon dioxide (CO(2)) production, and primary and ultimate half-lives. Interpretation of the applicability domain of models is also discussed.