Objective and subjective reduction of cellulite volume using a localized vibrational massage device in a 24-week randomized intra-individual single-blind regression study.

Cellulite occurs in females and is a common condition of altered connective tissue matrix and increased adipogenicity with visible dimples and orange-peel appearance on the skins surface. Whilst advancements in methods continue to help our understanding, attempts to correct the appearance of cellulite topically have yielded limited success. Various kinds of non-invasive body contouring methods such as whole body vibration have been reported with demonstrable visible improvements in the cellulite condition. The aim of this study was to evaluate volume reduction and improvement of the visible appearance of cellulite as judged both objectively (AEVA-HE phase-shift 3-D fringe projection, macrophotography image grading) and subjectively (questionnaires) after application of a hand-held localized vibrational device over 24-weeks. The study was conducted on 40 healthy female volunteers who were instructed how to use the device on defined areas of cellulite of the outside and rear of the thighs (iliotibial band, and over biceps femoris region respectively). The initial 12 weeks of continuous massage application of the study were followed by a 12 week phase in which volunteers were split into 2 subgroups - one for assessment of regression effects and one for continuous application effects. AEVA (skin surface volume) measurements of cellulite-related dimples correlated with questionnaires and visual image evaluation scoring, in that in the iliotibial region cellulite was significantly reduced at 12 weeks. In the regression subgroup cellulite returned to initial values soon after cessation of treatment, whereas in the continuous application subgroup, cellulite remained diminished. The effect of this device to reduce cellulite as observed in this study proves that continuous use of vibrational massage is beneficial to mitigate visible signs of cellulite.

La cellulite se produit chez les femmes et est un état d'altération courant de la matrice du tissu conjonctif et d'une adipogenèse accrue se manifestant par l´apparition des fossettes visibles et un aspect de peau d'orange à la surface de la peau. Bien que les progrès des méthodes approfondissent notre compréhension, les tentatives de corrections de l'apparence de la cellulite par voie topique ont produit des résultats limités. Divers types de méthodes non invasives du body contouring, telle que la vibration entière de celui-ci, ont été rapportées, et ce, avec des améliorations visibles et démontrables de l'état de la cellulite.L'objectif de cette étude était d'évaluer la réduction du volume et l'amélioration de l'apparence visible de la cellulite, comme jugées à la fois objectivement (AEVA-HE phase-shift 3-D projection de franges, gradation d'images par macrophotographie) et subjectivement (questionnaires) après application localisée d'un appareil vibratoire manuel pendant 24 semaines. L'étude a été menée sur 40 femmes volontaires en bonne santé qui ont reçu des instructions à-propos de comment utiliser l'appareil sur des zones externes et bien définies de la cellulite et à l'arrière des cuisses (bande iliotibiale et sur la région du biceps fémoral respectivement). Les 12 premières semaines d'application de massage continue ont été suivies d'une phase de 12 semaines au cours de laquelle les volontaires ont été divisés en deux sous-groupes - un pour l'évaluation des effets de régression et un pour l'évaluation des effets de l'application continue.Les mesures AEVA (volume de la surface de la peau) des capitons liés à la cellulite corrélées avec les questionnaires et le score visuel d'évaluation d'images, dans laquelle, la cellulite dans la région iliotibiale, était significativement réduite à 12 semaines. Dans la régression du sous-groupe, la cellulite est revenue aux valeurs initiales peu après l'arrêt du traitement, tandis que dans le sous-groupe d'application continue, la cellulite est restée diminuée. L'effet de ce dispositif pour réduire la cellulite tel qu'observé dans cette étude prouve que l'utilisation continue du massage vibratoire est bénéfique pour mitiger les signes visibles de la cellulite.

Matrix metalloproteinase 19 processes the laminin 5 gamma 2 chain and induces epithelial cell migration.

In this study we analyzed the proteolytic activity of MMP-19 and its impact on keratinocyte migration. In the HaCaT keratinocyte cell line overexpressing wild-type MMP-19 (HaCaT-WT), transmigration through fibrin and type IV collagen matrices was significantly increased compared to cells harboring a catalytically inactive mutant (HaCaT-EA). Studying the expression of MMP-19 in early stages of squamous cell cancer (SCC), we found co-localization of MMP-19 and laminin 5 at the invading tumor front but not in suprabasal epidermis of the tumor. Examination of laminin 5 processing revealed increased processing of the gamma2 chain in the medium and matrix of HaCaT-WT cells and degradation by recombinant human MMP-19 to 105-kDa and 80-kDa fragments. Parental HaCaT grown on the matrix of HaCaT-WT and HaCaT-EA cells displayed differential tyrosine phosphorylation. Using integrin blocking and stimulating antibodies we could attribute these differences to a shift from beta4-integrin-dependent signaling on the HaCaT-EA matrix toward alpha3-integrin-dependent signaling on the HaCaT-WT matrix. As a consequence, parental HaCaT showed increased migration on the matrix of HaCaT-WT cells. These data suggest that the MMP-19-dependent processing of the gamma2 chains leads to the integrin switch favoring epithelial migration and that MMP-19 actively participates in the early stages of SCC invasion.

Activity of MMP-19 inhibits capillary-like formation due to processing of nidogen-1.

Matrix metalloproteinase 19 (MMP-19) is able to process various proteins of the basement membrane. To investigate the impact of MMP-19 activity on endothelial cells in the context of tumor extracellular matrix (ECM), we treated Matrigel matrix with an active recombinant MMP-19 and analyzed its effect on capillary-like formation. Human microvascular endothelial cells (HMEC-1) could not form capillary-like formation on Matrigel treated with recombinant MMP-19. Analyzing the Matrigel proteins, we found that MMP-19 preferentially cleaved nidogen-1. The cleavage site of nidogen-1 was mapped to Thr867-Leu868. This cleavage separates the G3 globular domain containing the binding site for the gamma1 chain of laminin-1 and collagen IV and thus abolishes the capacity of nidogen-1 to cross-link ECM proteins. Anti-nidogen antibodies directed against the G3 domain of nidogen-1 inhibited the capillary-like structure formation to a similar extent as MMP-19. Since nidogen-1 is thought to stabilize microvessels, MMP-19 might be one of the enzymes that interferes with stabilization or maturation of nascent vasculature.

Differential effects of nonsteroidal antiinflammatory drugs on the IL-1 altered expression of plasminogen activators and plasminogen activator inhibitor-1 by articular chondrocytes.

OBJECTIVE: To determine the in vitro effects of several nonsteroidal antiinflammatory drugs on the IL-1 altered expression and activity of tPA, uPA and PAI-1 by articular chondrocytes. METHODS: Bovine chondrocytes were cultured in alginate gel beads. Cells were treated with IL-1alpha in the presence or absence of drugs at various concentrations. Expression of mRNA for the plasminogen activators (uPA and tPA) and their inhibitor (PAI-1) were analyzed by RT-PCR-ELISA. The protein content of PAI-1 in culture media was deter mined by ELISA. PA activity was measured by a functional assay. RESULTS: All tested NSAIDs dose dependently inhibited the IL-1 induced mRNA expression of tPA, whereas only indomethacin and tiaprofenic acid were also able to reduce the expression of uPA. Expression of PAI-1 was elevated by IL-1 without an accompanying increase in secreted amounts of the inhibitor. Indomethacin, naproxen and tiaprofenic acid stimulated the release of PAI-1 into culture media, whereas meloxicam also induced expression of PAI-1 above IL-1 stimulated levels. CONCLUSION: In conclusion, our studies indicate that NSAIDs preferentially inhibit tPA expression by bovine articular chondrocytes. By increasing the production of PAI-1 at therapeutical concentrations meloxicam could reduce PA activity, whereas the other NSAIDs tested mainly enhanced the release of this inhibitor from the extracellular matrix. In how far this would affect the enzyme-inhibitor balance within cartilage has to be determined in further studies.

Effects of non-steroidal antiinflammatory drugs and dexamethasone on the activity and expression of matrix metalloproteinase-1, matrix metalloproteinase-3 and tissue inhibitor of metalloproteinases-1 by bovine articular chondrocytes.

OBJECTIVE: To determine the in-vitro effects of several non-steroidal antiinflammatory drugs and the glucocorticoid dexamethasone on the IL-1 altered expression and activity of MMP-1, MMP-3 and TIMP-1 by bovine articular chondrocytes. DESIGN: Bovine chondrocytes were cultured in alginate gel beads. Cells were treated with IL-1alpha in the presence of vehicle or drugs at various concentrations. After 48 h mRNA expression of MMP-1, MMP-3, and of the tissue inhibitor of metalloproteinases (TIMP-1) was analysed by RT-PCR-ELISA. The protein synthesis of TIMP-1 and MMP-3 was determined by immunoprecipitation. The activity of enzymes and inhibitors was measured by functional assays. RESULTS: IL-1 increased the expression and activity of MMPs. In contrast, TIMP activity remained unchanged although TIMP-1 expression was down-regulated. All tested NSAIDs and dexamethasone inhibited collagenase activity induced by IL-1. Transcript levels of MMP-1, however, were only reduced by indomethacin, meloxicam, naproxen and dexamethasone. Proteoglycanase activity was only reduced by indomethacin, meloxicam and dexamethasone. These effects were pre-translational as confirmed by immunoprecipitation. The IL-1 decreased expression of TIMP-1 was further reduced by dexamethasone, which resulted in a significant loss of TIMP activity. No effects on TIMP activity or TIMP-1 biosynthesis were observed after treatment of chondrocytes with NSAIDs. CONCLUSION: Our studies clearly demonstrate that marked differences exist between individual NSAIDs with respect to their ability to modulate the imbalance between proteases and inhibitors during OA and RA, suggesting that the respective modes of action are independent of the inhibition of cyclooxygenases. Due to their co-regulation of MMPs and TIMP(s) glucocorticoids should be carefully studied for their overall effect on ECM proteolysis.

Effects of tetracyclines on the production of matrix metalloproteinases and plasminogen activators as well as of their natural inhibitors, tissue inhibitor of metalloproteinases-1 and plasminogen activator inhibitor-1.

OBJECTIVE: Evaluation of tetracycline effects on the expression of MMP-1, MMP-3, tissue inhibitor(s) of metalloproteinase-1 (TIMP-1), plasminogen activators (PA), and PA inhibitor-1, which are all involved in the ultimate regulation of MMP activity could provide new insight into how tetracyclines achieve their cartilage preserving effects. MATERIALS AND METHODS: We used bovine articular chondrocytes cultured in alginate gel beads for our studies which were initially treated with 10 microM tetracyclines in the presence of IL-1. Only significant effects were studied at additional concentrations. Expression of mRNA was analyzed by RT-PCR-ELISA. The activity of enzymes and TIMP was measured by functional assays; whereas, the level of PAI-1 was determined by ELISA. RESULTS: Treating chondrocytes with IL-1 induced the expression of MMPs and downregulated TIMP-1 but stimulated both the expression of PAs and PAI-1. When tested at 10 microM only minocycline reduced collagenase activity and expression of MMP-1. Further pharmacokinetic analysis revealed IC50 values of 26 microM and 16 microM for the inhibition of collagenase activity and mRNA expression, respectively. Production of MMP-3 was only decreased by tetracycline (IC50 = 45.4 microM). No effects of tetracyclines could be observed on proteoglycan degradation, TIMP activity and the production of PAs, PAI-1, and TIMP-1. CONCLUSIONS: We conclude that the inhibition of MMPs by tetracyclines occurs mainly via down-regulation of the respective gene expression.

Minocycline inhibits the production of inducible nitric oxide synthase in articular chondrocytes.

OBJECTIVE: To determine the in vitro effects of tetracyclines and nonsteroidal antiiflammatory drugs on interleukin 1alpha (IL-1alpha) induced NO production and biosynthesis of inducible NO synthase (iNOS) by articular chondrocytes. METHODS: Bovine chondrocytes were cultured in alginate beads. Cells were treated with IL-lalpha in the presence or absence of drugs at various concentrations. Expression of mRNA for iNOS was analyzed by reverse transcription polymerase chain reaction-ELISA. Protein synthesis of iNOS was determined by immunoprecipitation. NO production was taken as a measure for the activity of the enzyme. RESULTS: Minocycline dose dependently reduced IL-1 stimulated NO production by inhibition of the mRNA expression (IC50 = 69.9 microM) and protein synthesis (IC50 = 37.11 microM) of iNOS. Diclofenac-Na at a concentration of 10 microM only weakly reduced nitrite accumulation and mRNA expression of iNOS. No effects were observed for tetracycline, doxycycline, and meloxicam. CONCLUSION: Inhibition of iNOS in articular chondrocytes may be a new mechanism by which minocycline could exert its beneficial effects in the treatment of joint diseases.

The murine ortholog of matrix metalloproteinase 19: its cloning, gene organization, and expression.

We have isolated a murine cDNA orthologous to the human matrix metalloproteinase 19 (hMMP-19). The murine MMP-19 cDNA was amplified by RT-PCR using specific primers whose DNA sequences were derived from both murine MMP-19 genomic DNA and partial cDNA sequences. The murine MMP-19 (mMMP-19) is 79% identical to the human ortholog and encodes a protein of 527 amino acids with a deduced molecular mass of 59.1kDa. Analyzing the exon/intron junctions we revealed that the murine MMP-19 gene consists of nine exons and eight introns, and thus differs from the gene organization of other matrix metalloproteinases. Furthermore, a 587bp fragment of the mMMP-19 promoter containing a TATA box and an AP-1 binding motif was cloned, and 3.3kb transcripts of the MMP-19 gene were identified in liver, kidney, spleen, and colon. Finally, immunostaining of murine heart cryosections showed that mMMP-19, like its human counterpart, is expressed in the arterial tunica media of large blood vessels. By cloning mMMP-19 and unraveling its genomic structure, we have obtained valuable information for further study of the function of this MMP in vivo.

Focal liver disease: neural network-aided diagnosis based on clinical and laboratory data.

OBJECTIVES: The purpose of this study was to evaluate the use of clinical and laboratory data in the diagnosis of benign or malignant focal liver disease. METHODS: Diagnosis was made by artificial neural network (NN), a system of simple computing units connected in a specific structural network. Seven clinical and laboratory variables were retrospectively studied in 172 patients with a liver mass (93 benign, 79 malignant) detected with ultrasound. The diagnostic efficacy of NN was compared with a score based on the logistic regression model (Beaujon score). RESULTS: Although the sensitivity of the Beaujon score and the neural network was similar (4 malignant tumors inversely classified), neural network-aided diagnosis was characterized by higher specificity and accuracy (respectively 98.9% vs 82.5%, P < 0.001, and 97.1% vs 88.4%, P < 0.002). CONCLUSION: In patients with a hepatic mass, neural network is a valuable method for differentiating malignant and benign tumors.

[Preliminary studies of the occurrence of Ixodes ricinus (Acari: Ixodida: Ixodidae) in the areas of the Cracow-Czestochowa highland at various stages of human industrial activities]. / Wstepne badania nad wystepowaniem Ixodes ricinus (Acari: Ixodida: Ixodidae) na obszarach Jury Krakowsko-Czestochowskiej w róznym stopniu zmienionych antropopresja.

In the years 1987-88 the density of the population of I. ricinus on the territory of Kraków-Czestochowa Highland including the Ojców National Park was studied. The method of collection of ticks from 100 m2 fields was applied. On the spite of similar ecological conditions there was marked variation in the population density of I. ricinus. It decreased from East to West, where territories are under the strong pressure of the Upper Silesia and Olkusz industry.