Expression of the semicarbazide-sensitive amine oxidase in articular cartilage: its role in terminal differentiation of chondrocytes in rat and human.

OBJECTIVE: Semicarbazide-sensitive amine oxidase (SSAO) catalyzes the oxidation of primary amines into ammonia and reactive species (hydrogen peroxide, aldehydes). It is highly expressed in mammalian tissues, especially in vascular smooth muscle cells and adipocytes, where it plays a role in cell differentiation and glucose transport. The study aims at characterizing the expression and the activity of SSAO in rat and human articular cartilage of the knee, and to investigate its potential role in chondrocyte terminal differentiation. DESIGN: SSAO expression was examined by immunohistochemistry and western blot. Enzyme activity was measured using radiolabeled benzylamine as a substrate. Primary cell cultures of rat chondrocytes were treated for 21 days by a specific SSAO inhibitor, LJP 1586. Terminal chondrocyte differentiation markers were quantified by RT-qPCR. The basal and IL1ß-stimulated glucose transport was monitored by the entrance of (3)[H]2-deoxyglucose in chondrocytes. RESULTS: SSAO was expressed in chondrocytes of rat and human articular cartilage. SSAO expression was significantly enhanced during the hypertrophic differentiation of chondrocytes characterized by an increase in MMP13 and in alkaline phosphatase (ALP) expressions. SSAO inhibition delayed the late stage of chondrocyte differentiation without cell survival alteration and diminished the basal and IL1ß-stimulated glucose transport. Interestingly, SSAO activity was strongly increased in human osteoarthritic cartilage. CONCLUSIONS: SSAO was expressed as an active form in rat and human cartilage. The results suggest the involvement of SSAO in rat chondrocyte terminal differentiation via a modulation of the glucose transport. In man, the increased SSAO activity detected in osteoarthritic patients may trigger hypertrophy and cartilage degeneration.

A new case of sodium-dependent multivitamin transporter defect occurring as a life-threatening condition responsive to early vitamin supplementation and literature review.

BACKGROUND: Biallelic pathogenic variants in SLC5A6 resulting in sodium-dependent multivitamin transporter (SMVT) defect have recently been described as a vitamin-responsive inborn error of metabolism mimicking biotinidase deficiency. To our knowledge, only 16 patients have been reported so far with various clinical phenotypes such as neuropathy and other neurologic impairments, gastro-intestinal dysfunction and failure to thrive, osteopenia, immunodeficiency, metabolic acidosis, hypoglycemia, and recently severe cardiac symptoms. METHODS: We describe a case report of a 5-month-old girl presenting two recurrent episodes of metabolic decompensation and massive cardiac failure in the course of an infectious disease. We compare clinical, biological, and genetic findings of this patient to previous literature collected from Pubmed database (keywords: Sodium-dependent multivitamin transporter (SMVT), SMVT defect/disorder/deficiency, SLC5A6 gene/mutation). RESULTS: We highlight the life-threatening presentation of this disease, the stagnation of psychomotor development, the severe and persistent hypogammaglobulinemia, and additionally, the successful clinical response on early vitamin supplementation (biotin 15 mg a day and pantothenic acid 100 mg a day). Metabolic assessment showed a persistent increase of urinary 3-hydroxyisovaleric acid (3-HIA) as previously reported in this disease in literature. CONCLUSION: SMVT deficiency is a vitamin-responsive inborn error of metabolism that can lead to a wide range of symptoms. Increased and isolated excretion of urinary 3-hydroxyisovaleric acid may suggest, in the absence of markedly reduced biotinidase activity, a SMVT deficiency. Prompt supplementation with high doses of biotin and pantothenic acid should be initiated while awaiting results of SLC5A6 sequencing as this condition may be life-threatening.

Harmonization of strategies for the validation of quantitative analytical procedures: a SFSTP proposal part IV. Examples of application.

A harmonized approach for the validation of analytical methods based on accuracy profile was introduced by a SFSTP commission on the validation of analytical procedure. This fourth and last document aims at illustrating this methodology and the statistics used. Therefore the validation of real case methods are proposed such as methods for the quality control of drugs, for the quantitation of impurities in drug substances, for bioanalysis or for the determination of nutriments. Furthermore, different types of analytical methods are used in order to demonstrate the applicability of the proposed approach to a wide range of methods such as liquid chromatography (LC-UV, LC-MS), spectrophotometry or ELISA.

Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--part III.

In the first two documents [Ph. Hubert, J.J. Nguyen-Huu, B. Boulanger, E. Chapuzet, P. Chiap, N. Cohen, P.A. Compagnon, W. Dewé, M. Feinberg, M. Lallier, M. Laurentie, N. Mercier, G. Muzard, C. Nivet, L. Valat, J. Pharm. Biomed. Anal. 36 (2004) 579-586; Ph. Hubert, J.J. Nguyen-Huu, B. Boulanger, E. Chapuzet, P. Chiap, N. Cohen, P.A. Compagnon, W. Dewé, M. Feinberg, M. Lallier, M. Laurentie, N. Mercier, G. Muzard, C. Nivet, L. Valat, E. Rozet, J. Pharm. Biomed. Anal., in press], a recent SFSTP Commission on the validation of analytical procedure has introduced a harmonized approach for the validation of analytical procedures. In order to complete this guide, the statistical methodology allowing to correctly conclude about the validity of a procedure is proposed in this third part of the guide. Indeed all the steps to obtain the decision tool namely the accuracy profile are described and illustrated step by step by a numerical example. This tool, based on the concept of total error (bias+standard deviation) build with a beta-expectation tolerance interval, allows to easily take the right decision and simultaneously minimizing the risk of the future use of the analytical procedure.

Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--part II.

As reported in a previous paper, the main objective of the new commission of the Société Française des Sciences et Techniques Pharmaceutiques (SFSTP) was the harmonisation of approaches for the validation of quantitative analytical procedures. In a series of meetings, members of this Commission have first tried to review the objectives of analytical methods and the objectives of validation methods and to recommend the use of two-sided beta-expectation tolerance intervals for total error of validation samples (accuracy profile) in the acceptance/rejection of analytical method in validation phase. In the context of the harmonization, the other objectives were: (i) to propose a consensus on the norms usually recognized, while widely incorporating the ISO terminology; (ii) to recommend to validate the analytical procedure accordingly to the way it will be used in routine; (iii) to elaborate a rational, practical and statistically reliable strategy to assure the quality of the analytical results generated. This strategy has been formalised in a guide and the three latter objectives made by the Commission are summarised in the present paper which is the second part of summary report of the SFSTP commission. The SFSTP guide has been produced to help analysts to validate their analytical methods. It is the result of a consensus between professionals having expertise in analytical and/or statistical fields. The suggestions presented in this paper should therefore help the analyst to design and perform the minimum number validation experiments needed to obtain all the required information to establish and demonstrate the reliability of its analytical procedure.

A robust viologen and Mn-based porous coordination polymer with two types of Lewis acid sites providing high affinity for H2O, CO2 and NH3.

A novel porous coordination polymer [Mn(pc3)(H2O)2]·xH2O (3 < x < 4) is synthesized in water at pH = 7 using the anionic viologen-carboxylate ligand 4,4'-bipyridinium,1,1'-bis-(2,4-dicarboxyphenyl) (pc32-). Dehydration of the material results in the formation of open pores containing two types of accessible Lewis acid sites: exposed Mn2+ cations and N+ atoms of viologen units. Due to this property the PCP shows high affinity and capacity in the adsorption of H2O, CO2 and NH3. Despite the presence of strong adsorption sites this material is stable in liquid water and in gaseous NH3.

Significance of bone-marrow scintigraphy in aplastic anemia: concise communication.

Tc-99m colloid and In-111 transferrin were used in a semiquantitative scintigraphic study of bone-marrow activity in 76 patients with aplastic anemia, the majority of which were severe cases. The results are compared with other known prognostic parameters and with a predictive index formulated from a prior multi-parametric analysis performed in 352 cases. In 47 cases parallel abnormality of Tc and In uptakes was noted and was well correlated with other prognostic factors. Indium uptake is apparently a good indicator of the severity of aplasia; extension of active erythroid tissue, demonstrated with this method, is correlated with prognosis. In nine cases, excessive In uptake is explained by dyserythropoiesis associated with granulo- and thrombocytopenia (Fanconi's anemia in most cases). In 20 of our patients, TcSC uptake was excessive compared with that of In and with other prognostic factors. Statistically, this phenomenon carries an unfavorable prognosis but its physiological meaning remains to be defined.

Temperature-dependent sex determination and gonadal differentiation in reptiles.

In many reptile species, sexual differentiation of gonads is sensitive to temperature (temperature-dependent sex determination, TSD) during a critical period of embryonic development (thermosensitive period, TSP). Experiments carried out with different models including turtles, crocodilians and lizards have demonstrated the implication of estrogens and the key role played by aromatase (the enzyme complex that converts androgens to estrogens) in ovary differentiation during TSP and in maintenance of the ovarian structure after TSP. In some of these experiments, the occurrence of various degrees of gonadal intersexuality is related to weak differences in aromatase activity, suggesting subtle regulations of the aromatase gene at the transcription level. Temperature could intervene in these regulations. Studies presently under way deal with cloning (cDNAs) and expression (mRNAs) of genes that have been shown, or are expected, to be involved in gonadal formation and/or differentiation in mammals. Preliminary results show that homologues of the WT1, SF1, SOX9, DAX1 and AMH genes exist in TSD reptiles. However, the expression patterns of these genes during gonadal differentiation may be different between mammals and TSD reptiles and also between different reptile species. How these genes could interact with aromatase is being examined.

Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--Part I.

This paper is the first part of a summary report of a new commission of the Société Française des Sciences et Techniques Pharmaceutiques (SFSTP). The main objective of this commission was the harmonization of approaches for the validation of quantitative analytical procedures. Indeed, the principle of the validation of theses procedures is today widely spread in all the domains of activities where measurements are made. Nevertheless, this simple question of acceptability or not of an analytical procedure for a given application, remains incompletely determined in several cases despite the various regulations relating to the good practices (GLP, GMP, ...) and other documents of normative character (ISO, ICH, FDA, ...). There are many official documents describing the criteria of validation to be tested, but they do not propose any experimental protocol and limit themselves most often to the general concepts. For those reasons, two previous SFSTP commissions elaborated validation guides to concretely help the industrial scientists in charge of drug development to apply those regulatory recommendations. If these two first guides widely contributed to the use and progress of analytical validations, they present, nevertheless, weaknesses regarding the conclusions of the performed statistical tests and the decisions to be made with respect to the acceptance limits defined by the use of an analytical procedure. The present paper proposes to review even the bases of the analytical validation for developing harmonized approach, by distinguishing notably the diagnosis rules and the decision rules. This latter rule is based on the use of the accuracy profile, uses the notion of total error and allows to simplify the approach of the validation of an analytical procedure while checking the associated risk to its usage. Thanks to this novel validation approach, it is possible to unambiguously demonstrate the fitness for purpose of a new method as stated in all regulatory documents.

Modeling of transient deformation of piezoelectric ceramics [US NDE].

A method based on a finite-element analysis that permits the calculation and the visualization of deformations of ultrasonic transducers, in different configurations is presented. The method uses a Fourier series synthesis giving deformation of the free surface and electrical admittance for wide frequency bands by the use of finite-element procedures previously developed at Universite de Technologie de Compiegne (UTC). The method was originally developed for the monochromatic case. The authors report on the adaptation and validation of the method for the multifrequency excitation case, which is a more realistic approach used in the field of ultrasonic NDE. Several geometries were tested. In spite of the limitations of the method, due to the calculation requirements, the results show the role of the radial displacement behavior usually neglected in monodimensional analysis.

Fractures of the distal humerus operated on using the Lambda® plate: report of 75 cases at 9.5 years follow-up.

INTRODUCTION: The study sought to assess the functional results and complications of osteosynthesis using the Lambda(®) plate (Zimmer, Étupes, France) in treating fracture of the distal extremity of the humerus. MATERIALS AND METHODS: The initial series comprised 115 patients (116 fractures), treated between 1992 and 2008. Forty-one (mainly foreigners) were lost to follow-up. The final series thus comprised 74 patients (75 fractures): 44 female and 30 male, with a mean age of 46±23 years (range, 16-95 years), 22 (29%) being aged 65 years or over. According to AO classification, 22 of the fractures of the distal extremity of the humerus (29%) were diaphyseal-metaphyseal, corresponding to a particular grade of type A2, 12 (16%) were type A2 or A3, six (8%) type B, and 35 (47%) type C. Eight were open fractures: seven grade 1 and one grade IIa on Gustilo's classification. The Lambda(®) plate is an inverted Y shape, with a stem of up to ten holes and sectile arms that can be remodeled to adapt perfectly to the type of fracture and the shape of the distal end of the humerus. A posterior approach was used in all cases: in 26 cases, an extra-articular transolecranal approach and otherwise a transtricipital approach, either vertical for diaphyseal-metaphyseal fracture or inverted V for type-C fracture. RESULTS: Mean follow-up was 115±64 months (range, 24-227 months). There were no cases of infection, non-union of olecranial osteotomy or disassembly of the internal fixation; there was, on the other hand, one case of non-union of the lateral condyle and one of the distal extremity of the humerus, two cases of dysesthesia in the ulnar nerve territory and one in the radial nerve territory (following preoperative paralysis), and four cases of stiffness requiring surgical arthrolysis between 6 and 12 months postoperatively. At follow-up, mean active elbow flexion was 133°±13° (range, 90°-150°) and active extension -12°±14° (range, -45°-0°). Mean elbow flexion range of motion exceeded 100° in 58 patients (77%), was between 50° and 100° in 16 (21%) and was less than 50° in one. Mean Mayo Elbow Performance Score (MEPS) was 97±7 points (range, 40-100), and mean Quick DASH Score (converted as a score out of 100) was 10±18 (range, 0-54). There were 67 excellent results (MEPS, 90-100 points), five good (75-89), two moderate and one poor. The 35 type-C fractures displayed no significant differences from the series as a whole (P=1.24 for MEPS). CONCLUSION: Osteosynthesis using the Lambda(®) plate gave excellent medium-term results in terms of both fixation stability and recovery of elbow function after fracture of the distal extremity of the humerus, even in elderly osteoporotic patients. LEVEL OF EVIDENCE: Level IV: retrospective study.

Medial facet patelloplasty in patellar instability associated with patellofemoral dysplasia: a report of 26 cases.

INTRODUCTION: Numerous procedures may be used in the surgical treatment of patellar instability. We have noticed that certain types of patellar instability result in a bulge (or protrusion) of the distal medial facet of the patella which can hinder recentering in the trochlear groove. HYPOTHESIS: In certain patellofemoral dysplasias, trimming down of this bulge (patelloplasty) as long as it is associated with a "à la carte surgery" program, can improve stability, centering and lateral patellar tilt. PATIENTS AND METHODS: This retrospective series included 23 patients (26 knees), 13 women and 10 men, mean age 25.9±9.01 years old (15-52) operated between 1997-2008. Patellar dislocation had occurred at least once in all knees, and at least twice in 16 knees. The mean preoperative Kujala score was 79.1±6.1 points (68-91). There was a bulge on the medial facet of the patella in all cases, in particular on skyline views associated with the usual criteria for patellofemoral dysplasia. Patelloplasty was associated in all cases with resection of the lateral patellar retinaculum and anteromedialization of the tibial tubercle (7 were lowered) and four Albee trochleoplasties. RESULTS: Recurrent dislocation occurred in one case (4.7%). Nineteen patients (22 knees) were followed up for a mean 7.53±3.27 years (2-13 years). The mean postoperative Kujala score was 91.8±7.9 points (70-100) and the subjective results in 19/22 (86.5%) knees were satisfactory or very satisfactory. On skyline views, 21 patellae (95.5%) were well centered and one (4.5%) still presented with lateral tilt. There was no osteoarthritis in 15 knees (68%). CONCLUSION: In selected cases, patelloplasty of the medial facet of the patella has no particular morbidity at intermediate follow-up. Associated with other surgical procedures, the rate of satisfaction of patients is very high and especially encouraging. LEVEL OF EVIDENCE: Level IV, retrospective cohort study.

Posterior approach and dislocation rate: a 213 total hip replacements case-control study comparing the dual mobility cup with a conventional 28-mm metal head/polyethylene prosthesis.

INTRODUCTION: Dislocation is a frequent complication of total hip arthroplasties (THA) especially in older patients, especially when using a posterior approach. In these cases, dual mobility (DM) cups developed by Gilles Bousquet in 1975 can be indicated to reduce this complication risk. HYPOTHESIS: Dual mobility cups reduce the rate of dislocation in primary total hip arthroplasty using posterior approach in a single-surgeon series. AIM: Test this hypothesis in a controlled study to compare the rate of dislocation in primary total hip arthroplasties done in patients over 50 years old either with a dual mobility cup or a conventional metal-on-polyethylene 28-mm diameter head. PATIENTS AND METHODS: Two consecutive series of primary total hip replacements were performed by a single surgeon using a posterolateral approach. The piriformis tendon was left intact. The DM series included 105 patients who underwent arthroplasty between January 2005 and June 2007 with a dual mobility cup (60 women and 45 men, mean age 76.6±5.65 years old [53-93]). The control series (S series) included 108 patients who underwent arthroplasty (56 women and 52 men, mean age 74.2±5.9 years old [53-87]) with a conventional 28-mm polyethylene cup between January 2003 and June 2005. All hip replacements included a 28-mm metal-polyethylene cup and a 12-14-mm Morse taper. Both groups were comparable for gender, diagnosis, body mass index, type of anesthesia and ASA score distribution. All patients included in this series had a minimum follow-up of 1 year. RESULTS: There were no dislocations in the DM series and five early dislocations (before the third month) in the S series for a rate of 4.63%. Although the rate of dislocation was higher in the S series (4.63% vs 0%), the difference was barely significant (P=0.0597). DISCUSSION: This study comparing the incidence of dislocations after THA with conventional or dual mobility cups, shows that even using a posterior approach and in older patients, dual mobility cups increase stability with no postoperative dislocations. Although results are barely significant, a larger series should confirm the benefit of this implant. In this series, morbidity was not increased with dual mobility cups. LEVEL OF EVIDENCE: Level III: retrospective case-control study.

Limitation of adipose tissue enlargement in rats chronically treated with semicarbazide-sensitive amine oxidase and monoamine oxidase inhibitors.

Inhibition of semicarbazide-sensitive amine oxidases (SSAO) and monoamine oxidases (MAO) reduces fat deposition in obese rodents: chronic administration of the SSAO-inhibitor semicarbazide (S) in combination with pargyline (MAO-inhibitor) has been shown to reduce body weight gain in obese Zucker rats, while (E)-2-(4-fluorophenethyl)-3-fluoroallylamine, an SSAO- and MAO-B inhibitor, has been reported to limit weight gain in obese and diabetic mice. Our aim was to state whether such weight gain limitation could occur in non-obese, non-diabetic rats and to extend these observations to other amine oxidase inhibitors. Prolonged treatment of non-obese rats with a high dose of S (900 micromol kg(-1) day(-1)) reduced body weight gain and limited white adipose tissue enlargement. When chronically administered at a threefold lower dose, S also inhibited SSAO activity but not fat depot enlargement, suggesting that effects other than SSAO inhibition were involved in adipose tissue growth retardation. However, combined treatment of this lower dose of S with pargyline inhibited SSAO, MAO, energy intake, weight gain and fat deposition. Adipocytes from treated rats exhibited unchanged insulin responsiveness but impaired antilipolytic responses to amine oxidase substrates. Phenelzine clearly inhibited both MAO and SSAO when tested on adipocytes. Obese rats receiving phenelzine i.p. at 17 micromol kg(-1) day(-1) for 3 weeks, exhibited blunted MAO and SSAO activities in any tested tissue, diminished body weight gain and reduced intra-abdominal adipose tissue. Their adipocytes were less responsive to lipogenesis activation by tyramine or benzylamine. These observations suggest that SSAO inhibition is not sufficient to impair fat deposition. However, combined MAO and SSAO inhibition limits adiposity in non-obese as well as in obese rats.

Dating the demise: neandertal extinction and the establishment of modern humans in the southern Caucasus.

This paper considers the recent radiometric dating (14C-AMS, TL, ESR) of 76 late Middle and early Upper Paleolithic samples from Ortvale Klde Rockshelter, located in the Republic of Georgia. We present a critical evaluation of each date based on its stratigraphic and archaeological context, its pretreatment and contamination history, and its resulting accuracy and precision, the goal being to establish a sound chronology for the site. Only by systematically identifying aberrant dates within a data set and isolating them from further analysis can we hope to understand cultural and biological phenomena on an accurate temporal scale. Based on the strict discard protocol outlined here, we omit 25% of the dated samples from the analysis. The remaining data speak to the lengthy tenure of Neandertals in the region, but also to their relatively rapid demise and the establishment of modern human populations approximately 38-34 ka 14C BP (42-39 kacalBP(Hulu)). We compare these chronometric data with those from the neighboring sites of Bronze and Dzudzuana caves, as well as Mezmaiskaya Cave, located in the northern Caucasus. While the lack of key contextual information limit our ability to subject these other data sets to the same critical evaluation procedure, they provide the first interregional temporal assessment of the Middle to Upper Paleolithic "transition," the results of which suggest an initial expansion of modern humans into the southern Caucasus followed by expansion along the Black Sea coast and into the northern Caucasus.

[Polycythemias of young subjects (author's transl)]. / Polyglobulie des sujets jeunes. 86 observations.

During the last 12 years, 86 apparently primary polycythemias have been observed in subjects less than 40 years old. In twelve of them a non-suspected cause has been found (seven abnormalities of the hemoglobin function). In 43 cases, all males, the erythrocytosis (red cell volume more than 36 ml/kg) was pure and stable, without any complication; this "benign erythrocytosis" probably depicts extreme values of the normal distribution of the red cell mass. In 31 cases, the disease had criteria similar to those of polycythemia vera observed in older patients; in 12 of these cases, severe thrombo-embolic complications have been observed at the very beginning of the disease; however no further complication has been seen, when the patients were actively treated (phlebotomies or 32P); at the present time, no one of these cases did show transformation into acute leukemia, but the long-term prognosis (beyond 15 years) cannot yet been precisely drawn.