Reversal of TGF-β1 stimulation of α-smooth muscle actin and extracellular matrix components by cyclic AMP in Dupuytren's - derived fibroblasts

<p>Abstract</p> <p>Background</p> <p>Myofibroblasts, a derived subset of fibroblasts especially important in scar formation and wound contraction, have been found at elevated levels in affected Dupuytren's tissues. Transformation of fibroblasts to myofibroblasts is characterized by expression of alpha- smooth muscle actin (α-SMA) and increased production of extracellular matrix (ECM) components, both events of relevance to connective tissue remodeling. We propose that increasing the activation of the cyclic AMP (cAMP)/protein kinase A signaling pathway will inhibit transforming growth factor-beta1 (TGF-β₁)-induced ECM synthesis and myofibroblast formation and may provide a means to blunt fibrosis.</p> <p>Methods</p> <p>Fibroblasts derived from areas of Dupuytren's contracture cord (DC), from adjacent and phenotypically normal palmar fascia (PF), and from palmar fascia from patients undergoing carpal tunnel release (CTR; CT) were treated with TGF-β₁(2 ng/ml) and/or forskolin (10 μM) (a known stimulator of cAMP). Total RNA and protein extracted was subjected to real time RT-PCR and Western blot analysis.</p> <p>Results</p> <p>The basal mRNA expression levels of fibronectin- extra domain A (FN1-EDA), type I (COL1A2) and type III collagen (COL3A1), and connective tissue growth factor (CTGF) were all significantly increased in DC- and in PF-derived cells compared to CT-derived fibroblasts. The TGF-β₁stimulation of α-SMA, CTGF, COL1A2 and COL3A1 was greatly inhibited by concomitant treatment with forskolin, especially in DC-derived cells. In contrast, TGF-β₁stimulation of FN1-EDA showed similar levels of reduction with the addition of forskolin in all three cell types.</p> <p>Conclusion</p> <p>In sum, increasing cAMP levels show potential to inhibit the formation of myofibroblasts and accumulation of ECM components. Molecular agents that increase cAMP may therefore prove useful in mitigating DC progression or recurrence.</p

A new model dependency ratio for European cities.

Background: Sometimes referred to as ‘the demographic time bomb,’ the European Union, state and local governments are concerned about the impact of an ageing population on both sustainable economic development and the demand for health and social support services. Seeking to mitigate these pressures, the Organisation for Economic Co-operation and Development (OECD) has developed a policy framework Live Longer: Work Longer and the World Health Organization (WHO) has set a policy framework for Active Ageing which maintains that early life course interventions can reduce levels of disability and dependency in older age. The WHO European Healthy Cities Network (WHO-EHCN) promotes healthy urban planning to encourage healthy lifestyles and maintain older people as a resource in the workplace and to their communities. Our objective is to develop a new model dependency ratio (NMDR) for European cities which synthesises these three policy frameworks. Methods: Starting from the classic formulation of the dependency ratio (DR), which compares the 'dependent' population segments with the working-age or 'productive' segments, the model is developed in six stages, drawing on data from secondary European and national sources and from primary sources contained in Healthy Ageing Profiles of fifteen (WHO-EHCN) cities. Results: From an orthodox baseline, the second stage of modelling increases the DR by moving economically inactive people of working age from denominator to numerator. Thereafter, refinements introduced in stages three to six, gradually reduce the DR. Conclusions: The NMDR challenges the 'demographic time bomb' predicted by orthodox formulations and can be used as a tool by city decision makers

Inherited thrombophilia in infertile women: implication in unexplained infertility

Many studies evaluating a possible relationship between inherited thrombophilia and the etiology of unexplained infertility have been performed recently. No significant difference in the prevalence of three genetic mutations associated with the increased risk of thrombophilia (Factor V Leiden G1691A, prothrombin G20210A, and methylenetetrahydrofolate reductase [MTHFR] C677 T) was found in 100 infertile women with unexplained infertility when compared with 200 control fertile women without an infertility history

Homologous intrauterine insemination in controlled ovarian hyperstimulation cycles: a comparison among three different regimens

OBJECTIVE: The objective was to assess the efficacy of double intrauterine insemination (IUI) over a single periovulatory IUI in patients undergoing controlled ovarian hyperstimulation with low-dose recombinant follicle stimulating hormone (rFSH) combined with human chorionic gonadotropin (HCG). STUDY DESIGN: Ninety-four infertile women were randomly assigned to three groups; in group A (38 patients, 47 cycles) a single IUI was performed 36 h after HCG administration combined with timed intercourse the day of HCG administration; within group B (43 patients, 48 cycles) IUI alone was performed 36 h after HCG administration; in group C (39 patients, 43 cycles) a double IUI 12 and 36 h after HCG administration was performed. RESULTS: The mean age and the causes of infertility were similar between the three groups. The number of follicles greater than 15 mm on the day of HCG administration and the overall dose of rFSH required per cycle was not significantly different among the groups. The pregnancy rate (PR) per cycle and per patient was 14.9% and 18.4% in group A, 10.4% and 11.6% in group B, 20.9% and 23.1% in group C, respectively. There was no statistically significant difference in PR among the three groups. CONCLUSION: In rFSH/HCG cycles, two IUIs performed 12 and 36 h after HCG administration do not significantly improve pregnancy rates over a single insemination performed 36 h after HCG administration combined with or without timed intercourse the day of HCG administration

VOR gain calculation methods in video head impulse recordings.

BACKGROUND International consensus on best practices for calculating and reporting vestibular function is lacking. Quantitative vestibulo-ocular reflex (VOR) gain using a video head impulse test (HIT) device can be calculated by various methods. OBJECTIVE To compare different gain calculation methods and to analyze interactions between artifacts and calculation methods. METHODS We analyzed 1300 horizontal HIT traces from 26 patients with acute vestibular syndrome and calculated the ratio between eye and head velocity at specific time points (40 ms, 60 ms) after HIT onset ('velocity gain'), ratio of velocity slopes ('regression gain'), and ratio of area under the curves after de-saccading ('position gain'). RESULTS There was no mean difference between gain at 60 ms and position gain, both showing a significant correlation (r2 = 0.77, p < 0.001) for artifact-free recordings. All artifacts reduced high, normal-range gains modestly (range - 0.06 to - 0.11). The impact on abnormal, low gains was variable (depending on the artifact type) compared to artifact-free recordings. CONCLUSIONS There is no clear superiority of a single gain calculation method for video HIT testing. Artifacts cause small but significant reductions of measured VOR gains in HITs with higher, normal-range gains, regardless of calculation method. Artifacts in abnormal HITs with low gain increased measurement noise. A larger number of HITs should be performed to confirm abnormal results, regardless of calculation method

Freqüência da mutação 844ins68 do gene da cistationina beta-sintetase em pacientes com trombose venosa profunda

A hiper-homocisteinemia, resultante da deficiência na conversão da homocisteína em cistationina, constitui em fator de risco isolado para doenças vasculares. A mutação 844ins68 do gene da cistationina beta-sintetase é um fator adicional de risco para a trombose venosa profunda. O objetivo deste estudo foi avaliar a freqüência da mutação 844ins68 do gene da cistationina beta-sintetase em pacientes com trombose venosa profunda. Foram avaliados em estudo caso-controle 95 pacientes com trombose venosa profunda, a presença da mutação 844ins68 no éxon 8 do gene da cistationina beta-sintetase. Como critério de inclusão foi adotada a presença de trombose venosa profunda confirmada pelo dúplex ou flebografia. O grupo controle constituiu-se de 95 doadores de sangue, sem história familiar prévia de trombose venosa, com sexo, grupo étnico e idades pareados aos do grupo de estudo. Foram coletados 5 mL de sangue venoso com o uso de anticoagulante EDTA de cada participante. O DNA foi extraído dos leucócitos pelo método DTAB e CTAB. A detecção da mutação do gene foi realizada por amplificação de um segmento gênico por PCR, com iniciadores que flanqueiam a região de inserção e com revelação em gel de agarose a 2%, corado com brometo de etídio, sob luz UV. O fragmento correspondente ao alelo normal contém 184 pares de base e o correspondente ao alelo mutante, 252 pares de base. O teste exato de Fisher foi utilizado na análise dos resultados. A condição heterozigota para a mutação foi encontrada em 14,73% dos pacientes e em 3,1% dos indivíduos do grupo controle (p = 0,009). A freqüência do alelo mutante mostrou diferença significativa (p = 0,01), sendo 0,074 para os pacientes versus 0,016 para o grupo controle. Não foram encontrados casos de homozigose.Hyperhomocysteinemia, resulting from a deficiency in the conversion of homocysteine to cystathionine, constitutes an independent risk factor for vascular diseases. The 844ins68 mutation of the cystathionine-beta-synthetase gene is an additional risk factor for deep venous thrombosis. The aim of this study was to evaluate the frequency of the 844ins68 mutation of the cystathionine-beta-synthetase gene in deep venous thrombosis. In a case control study, 95 patients with deep venous thrombosis were evaluated in respect to the presence of the 844ins68 mutation on exon 8 of the cystathionine-beta-synthetase gene. The inclusion criterion included the presence of deep venous thrombosis confirmed by duplex or phlebography. A control group was formed consisting of 95 blood donors, without previous history of venous thrombosis with data such as gender, age and race similar to the study group. Five milliliters of venous blood were collected in EDTA anticoagulant from all the members of both groups. The DNA was extracted from the leukocytes by te DTAB and CTAB methods. Detection of the gene mutation was made by amplification by PCR, using primers for the insertion region and observed by 2% agarose gel electrophoresis using ethidium bromide stain. The fragment corresponding to the normal allele contains 184 base pairs and the mutant allele 252 base pairs. The Fisher exact test was utilized in the analysis of the results. Heterozygote individuals for the mutation were evidenced in 14.73% of the patients and in 3.1% of the control group (p-value = 0.009). The frequency of the mutant allele demonstrated a significant difference (0.074 for the patients versus 0.016 for the control group) (p-value = 0.01). No homozygotic individuals were found