Pulmonary arterial hypertension in interferonophaties: a case report and a review of the literature.

BACKGROUND: Pulmonary arterial hypertension consists in an increase of mean pulmonary arterial pressure (PAPm ≥ 25 mmHg), and may lead to right ventricular failure. Pulmonary arterial hypertension can arise in several disorders, encompassing inflammatory conditions and connective tissue diseases. The occurrence of pulmonary arterial hypertension has recently been reported in monogenic interferonopathies and in systemic lupus erythematosus, highlighting the pathogenic role of type I interferons and paving the way to therapies aimed at inhibiting interferon signaling. CASE: We describe a 17-year-old boy with DNase II deficiency, presenting a clinical picture with significant overlap with systemic lupus erythematosus. During treatment with the Janus kinase inhibitor ruxolitinib, he developed pulmonary arterial hypertension, raising the question whether it could represent a sign of insufficient disease control or a drug-related adverse event. The disease even worsened after drug withdrawal, but rapidly improved after starting the drug again at higher dosage. SUMMARY AND CONCLUSION: Pulmonary arterial hypertension can complicate type I interferonopathies. We propose that ruxolitinib was beneficial in this case, but the wider role of Janus kinase inhibitors for the treatment of pulmonary arterial hypertension is not clear. For this reason, a strict cardiologic evaluation must be part of the standard care of subjects with interferonopathies, especially when Janus kinase inhibitors are prescribed.

Long-term follow-up of nailfold videocapillaroscopic changes in dermatomyositis versus systemic sclerosis patients.

To identify nailfold videocapillaroscopy (NVC) changes in patients with dermatomyositis (DM) during a 3-year follow-up and to compare the NVC findings between DM and systemic sclerosis (SSc) patients at their first visit. Retrospective study of 24 DM and 24 SSc patients, matched for age and disease duration at first NVC. Capillaroscopic patterns/scores and clinical parameters had been yearly assessed. Nineteen out of 24 DM patients (79%) showed a NVC "scleroderma-like pattern." No statistically significant variation of all the capillaroscopic scores was observed during the 3-year follow-up. By comparing DM patients with or without anti-Jo-1 positivity, no statistically significant difference of the scores of the main capillary parameters was observed at baseline between the groups. Comparing at baseline DM with SSc patients, the giant capillary and microhemorrhage scores were significantly higher in SSc than those in DM patients (p = 0.04 and p = 0.05, respectively), while capillary density, ramification (abnormally shaped capillaries, expression of angiogenesis), and disorganization scores were higher in DM patients (p = 0.05, p = 0.002, p = 0.004, respectively). The absolute number of ramified capillaries was significantly higher in DM patients (p = 0.002), while the absolute capillary number was significantly higher in SSc patients (p = 0.05) at baseline. This pilot study demonstrates, for the first time, over long-term, that the capillaroscopic manifestations of DM persist in contrast to the progressive changes described in SSc patients, and the anti-Jo-1 positivity does not seem to modify the NVC pattern.

Retraction Note to: MIR221/MIR222-driven post-transcriptional regulation of P27KIP1 and P57KIP2 is crucial for high-glucose- and AGE-mediated vascular cell damage.

The authors are retracting this article [1]. Following publication, concerns were raised with respect to some of the western blots and the authors were asked to supply the original unmodified blots.

Correlations between blood perfusion and dermal thickness in different skin areas of systemic sclerosis patients.

OBJECTIVE: To identify possible correlations between skin blood perfusion (BP) and dermal thickness (DT) in different skin areas of systemic sclerosis (SSc) patients. METHODS: Sixty-two SSc patients, according to 2013 EULAR/ACR criteria, and 62 healthy subjects (CNT) were enrolled. Skin BP was analysed by laser speckle contrast analysis (LASCA) at the level of dorsum of the middle phalanx of the third fingers, dorsal aspect of the hands and zygoma. DT was assessed by both skin high frequency ultrasound (US) and modified Rodnan skin score (mRSS) in the same above reported areas. All patients were studied also by nailfold videocapillaroscopy (NVC) to assess the proper pattern of microvascular damage ("Early", "Active", or "Late"). RESULTS: At the level of finger dorsum a statistically significant negative correlation was observed in SSc patients between skin BP and both ultrasound-DT (p=0.0005 r=0.43) and mRSS (p=0.0007 r=0.42), but not at the level of hand dorsum and zygoma. No statistically significant correlation was present between skin BP and ultrasound-DT at any level in CNT. In detail, SSc patients, compared to CNT, showed a statistically significant lower BP only at level of fingers (median PU 72.6 vs 136.1 respectively, p<0.0001) and a statistically significant higher ultrasound-DT at the level of dorsum of 3th finger bilaterally (median mm 0.9 vs 0.7, p<0.0001), dorsum of hands (median mm 0.9 vs 0.7, p<0.0001) and zygoma (median mm 0.8 vs 0.7, p<0.0001). A significant positive correlation between ultrasound-DT and mRSS was observed in SSc patients at level of the three areas (dorsum of fingers p<0.0001 r=0.51; dorsum of hands p=0.03 r=0.27; zygoma p=0.0001 r=0.45). A progressive decrease of skin BP and increase of ultrasound-DT was found correlated with the progression of the severity of NVC patterns. CONCLUSIONS: This study demonstrates for the first time in SSc patients a significant inverse relationship between skin BP, measured by LASCA, and DT, evaluated by both US and mRSS, at the level of dorsum of the middle phalanx of the third fingers.

Sexual distress, sexual dysfunction and relationship quality in women with systemic sclerosis: correlation with clinical variables.

To assess the rate of sexual distress, sexual dysfunction and relationship quality and their association with clinical variables in women with systemic sclerosis (SSc), 102 sexually active women with SSc were recruited. Sexual distress, sexual dysfunction and dissatisfaction with relationship quality were investigated by Female Sexual Distress Scale Revised (FSDS-R), Female Sexual Function Index (FSFI) and Dyadic Adjustment Scale (DAS), respectively. The patients underwent medical examinations and nailfold videocapillaroscopy (NVC). Of the 102 patients, 37 (36%) reported sexual distress with FSDS-R score >11, 45 (44%) had sexual dysfunction with FSFI score <19 and 49 (48%) were not satisfied with relationship quality with DAS score <100. There was a negative correlation (p<0.001, R= -0.30) between FSDS-R and FSFI. No correlation was found between FSDS-R and DAS. FSFI showed a positive correlation with DAS (p<0.0001, R= 0.36). Age correlated negatively (p<0.05, R= -0.26) with FSFI, while FSDS-R and DAS did not correlate (p>0.05) with age. SSc women with digital ulcers (DU) had a reduction of FSFI and DAS compared with women without DU. In patients with late capillaroscopic pattern, mean value of FSFI was significantly lower than the other two capillaroscopic patterns. DAS decreased with progression of capillaroscopic damage. In a high percentage of women with SSc FSDS-R was increased, while FSFI and DAS were reduced. Age correlated negatively with FSFI, while skin score showed a negative correlation with DAS. Digital vascular damage negatively influenced FSFI and DAS.

A diabetic milieu promotes OCT4 and NANOG production in human visceral-derived adipose stem cells.

AIMS/HYPOTHESIS: Successful outcomes have been obtained by exploiting adipose-derived stem cells (ASCs) in regenerative medicine. NADPH oxidase (NOX)-generated reactive oxygen species (ROS) are known to control stem cell self-renewal. Several high glucose (HG)-mediated effects depend on NOX-generated ROS. In this study, we investigated whether, and how mechanistically, HG concentrations control ASC fate in patients with diabetes. METHODS: ASCs from the visceral adipose tissue of non-diabetic (N-ASCs) and diabetic participants (D-ASCs), identified by surface markers, were counted and evaluated for ROS generation and stem cell properties. Their ability to release soluble factors was assessed by BioPlex analysis. To reproduce an in vitro diabetic glucose milieu, N-ASCs were cultured in HG (25 mmol/l) or normal glucose (NG) concentration (5 mmol/l), as control. ASC pluripotency was assessed by in vitro study. The p47(phox) NOX subunit, AKT and octamer-binding transcription factor 4 (OCT4; also known as POU5F1) were knocked down by small-interfering RNA technology. Stem-cell features were evaluated by sphere cluster formation. RESULTS: The ASC number was higher in diabetic patients than in non-diabetic controls. Production of OCT4 and NANOG, stem-cell-specific transcription factors, was upregulated in D-ASCs compared with N-ASCs. Moreover, we found that ROS production and AKT activation drove D-ASC, but not N-ASC, secretion. When N-ASCs were cultured in vitro in the presence of HG, they also expressed OCT4/NANOG and formed spheres. By knock-down of the p47(phox) NOX subunit, AKT and OCT4 we demonstrated that NOX-generated ROS and their downstream signals are crucial for HG-mediated ASC de-differentiation and proinflammatory cytokine production. CONCLUSIONS/INTERPRETATION: We herein provide a rationale for exploiting D-ASCs in regenerative medicine and/or exploiting HG preconditioning to increase ASCs ex vivo.

MIR221/MIR222-driven post-transcriptional regulation of P27KIP1 and P57KIP2 is crucial for high-glucose- and AGE-mediated vascular cell damage.

AIMS/HYPOTHESIS: MicroRNAs (miRNAs) are a novel group of small non-coding RNAs that regulate gene expression at the post-transcriptional level and act on their target mRNAs in a tissue- and cell-type-specific manner. Herein, the relevance of MIR221/MIR222 in high-glucose- and AGE-mediated vascular damage was investigated. METHODS: Functional studies were performed using human mature endothelial cells and endothelial progenitor cells subjected to high glucose or AGE. Quantitative real-time amplification was performed to analyse MIR221/MIR222 expression in these experimental conditions. Luciferase assay was used to identify MIR221/MIR222 targets. Functional studies were performed in vitro and in vivo in mice using gain- and loss-of-function approaches. RESULTS: Using an in vivo mouse model we demonstrated that exposure to AGE and high glucose impaired vessel formation. Moreover, in vitro functional studies revealed that both high glucose and AGE inhibit cell-cycle progression by modulating the expression of P27KIP1 (also known as CDKN1B) and P57KIP2 (also known as CDKN1C), which encode cyclin-dependent kinase inhibitor 1B (p27, Kip1) (P27KIP1) and cyclin-dependent kinase inhibitor 1C (p57, Kip2) (P57KIP2), respectively. Crucial to AGE- and high-glucose-mediated cell-cycle arrest was the downregulation of MIR221/MIR222 expression. Luciferase assay showed that MIR221 and MIR222 specifically bind to the P27KIP1 and P57KIP2 mRNA 3'-untranslated regions, implicating P27KIP1 and P57KIP2 as MIR221/MIR222 targets. These results were confirmed by gain-of-function experiments in vitro, and by injecting mice with endothelial cells overexpressing MIR221 and MIR222. CONCLUSIONS/INTERPRETATION: We provide evidence that high-glucose- and AGE-induced inhibition of vascular cell proliferation is controlled by MIR221/MIR222-driven post-transcriptional regulation of P27KIP1 and P57KIP2. These data add further insight to the possible contribution of miRNAs in vascular damage mediated by a high-glucose environment.

Involvement of PPARs in Cell Proliferation and Apoptosis in Human Colon Cancer Specimens and in Normal and Cancer Cell Lines.

PPAR involvement in cell growth was investigated "in vivo" and "in vitro" and was correlated with cell proliferation and apoptotic death. "In vivo" PPARgamma and alpha were evaluated in colon cancer specimens and adjacent nonneoplastic colonic mucosa. PPARgamma increased in most cancer specimens versus mucosa, with a decrease in c-Myc and in PCNA proteins, suggesting that colon cancer growth is due to increased cell survival rather than increased proliferation. The prevalence of survival over proliferation was confirmed by Bcl-2 or Bcl-X(L) increase in cancer versus mucosa, and by decreased PPARalpha. "In vitro" PPARgamma and PPARalpha were evaluated in human tumor and normal cell lines, treated with natural or synthetic ligands. PPARgamma was involved in inhibiting cell proliferation with a decrease in c-Myc protein, whereas PPARalpha was involved in inducing apoptosis with modulation of Bcl-2 and Bad proteins. This involvement was confirmed using specific antagonists of two PPARs. Moreover, the results obtained on treating cell lines with PPAR ligands confirm observations in colon cancer: there is an inverse correlation between PPARalpha and Bcl-2 and between PPARgamma and c-Myc.

HMG-CoA reductase and PPARalpha are involved in clofibrate-induced apoptosis in human keratinocytes.

Contrasting data have been reported on the effects of clofibrate, a PPARalpha agonist and hypolipidemic drug. The carcinogenic and anti-apoptotic effects have been demonstrated especially in rodents in both "in vivo" and "in vitro" experiments. In contrast, in rat and human hepatoma cell lines, several reports have shown its concentration-dependent pro-apoptotic effect. No epidemiological data exist about its carcinogenetic effect in man. This study shows that clofibrate also induced apoptosis in a human non-tumour cell line, NCTC 2544, which shares the characteristic of proliferation with tumour cells. Both HMG-CoA reductase and PPARalpha were found to be involved in the signal transduction pathway inducing apoptosis, the former being the principal target: HMG-CoA reductase decreased and PPARalpha increased. Changes in HMG-CoA reductase expression caused activation of parameters leading to apoptosis via the mitochondria pathway. Clofibrate must be considered a pro-apoptotic molecule at concentrations of 0.25 mM and above: the effect is exercised not only on tumour cells but also on normal human proliferating cells. Clofibrate should thus be regarded as a potential drug to reduce the number of proliferating cells in pathological conditions.

Atrial fibrillation and mitral prolapse in a subject affected by Refetoff syndrome.

Inappropriate secretion of TSH (IST) refers to a heterogeneous group of syndromes in which patients show unsuppressed TSH levels in spite of high serum free thyroid hormone concentrations. It has been recognised that IST can be due to both thyroid hormone resistance (RTH) and pituitary TSH-secreting tumours. The former can be generalised (GRTH) or pituitary (PRTH) if the resistance is more severe in the pituitary than in the remaining tissues. This case report describes a peculiar coexistence of atrial fibrillation and mitral valve prolapse in a patient affected by generalized resistance to thyroid hormone. This finding is suggestive for a major and almost physiological sensitivity of the myocardium to the thyroid hormones activity which in the course of years may determine the modifications responsible for the pathologies described.

[Management of overweight and obese patient. New acquisition]. / Management del paziente in sovrappeso e obeso. Recenti acquisizioni.

Obesity is a chronic disease and prevalence and incidence are progressively increasing. Treatment of obesity is important to reduce mortality and associated diseases, like diabetes mellitus, hypertension, abnormal blood lipid levels, coronary heart disease, thromboembolic disease, cancer (endometrial, gallbladder, cervical, ovarian, breast, prostate and colorectal), polycystic ovary syndrome (PCOS), gallbladder disease, respiratory disease, arthritis, gout. Most of these pathologies profits by a modest weight loss (5-10%). A correct management of obesity should include integration of therapeutic strategies, that we have actually at disposal: diet, physical training, behaviour therapy, pharmacologic therapy and surgery. We should get together low-calorie and low-fat diet with behaviour change and physical training. Physical training induces a significant weight loss and reduces cardiovascular risks and insulin resistance. Orlistat, that reduces up to 30% lipid adsorption, is a valid remedy if with an adequate diet. A new drug, sibutramine, shows efficacy: it increases satiety and energy expenditure caused by thermogenesis in brown adipose tissue. Surgical approaches including some procedures, are indicated for great obesity (BMI >40).

Increase in class 2 aldehyde dehydrogenase expression by arachidonic acid in rat hepatoma cells.

Aldehyde dehydrogenase (ALDH) is a family of several isoenzymes important in cell defence against both exogenous and endogenous aldehydes. Compared with normal hepatocytes, in rat hepatoma cells the following changes in the expression of ALDH occur: cytosolic class 3 ALDH expression appears and mitochondrial class 2 ALDH decreases. In parallel with these changes, a decrease in the polyunsaturated fatty acid content in membrane phospholipids occurs. In the present study we demonstrated that restoring the levels of arachidonic acid in 7777 and JM2 rat hepatoma cell lines to those seen in hepatocytes decreases hepatoma cell growth, and increases class 2 ALDH activity. This latter effect appears to be due to an increased gene transcription of class 2 ALDH. To account for this increase, we examined whether peroxisome-proliferator-activated receptors (PPARs) or lipid peroxidation were involved. We demonstrated a stimulation of PPAR expression, which is different in the two hepatoma cell lines: in the 7777 cell line, there was an increase in PPAR alpha expression, whereas PPAR gamma expression increased in JM2 cells. We also found increased lipid peroxidation, but this increase became evident at a later stage when class 2 ALDH expression had already increased. In conclusion, arachidonic acid added to the culture medium of hepatoma cell lines is able to partially restore the normal phenotype of class 2 ALDH, in addition to a decrease in cell growth.

The effect of a novel irreversible inhibitor of aldehyde dehydrogenases 1 and 3 on tumour cell growth and death.

Aldehyde dehydrogenases (ALDHs) are a family of several isoenzymes expressed in various tissues and in all subcellular fractions. In some tumours, there is an increase of ALDH activity, especially that of class 1 and 3. The increase in the activity of these isoenzymes is correlated with cell growth and drug resistance shown by these cells. It has been observed that hepatoma cells expressing low ALDH3 activity are more susceptible to growth inhibition by low concentration of lipid peroxidation products than hepatoma cells expressing high ALDH3 activity. The products of lipid peroxidation are good substrates for ALDH, but when their intracellular levels are increased in hepatoma cells treated repeatedly with prooxidants, they inhibit ALDH3 and bring about growth inhibition or cell death. As a follow up to the work previously reported on S-methyl 4-amino-4-methylpent-2-ynethioate, a synthetic suicide inhibitor of ALDH1, which induced bcl2 overexpressing cells into apoptosis and exhibited an ED50 of 400 microM, a novel broad spectrum inhibitor of ALDH1 and ALDH3 was synthesised. This new compound (ATEM) is a suicide inhibitor of ALDH1, an irreversible inhibitor of ALDH3 and exhibits an ED50 of 10-25 microM on rat cultured hepatoma cells. Four hours after treatment with 25 microM ATEM, ALDH activity using benzaldehyde or propionaldehyde in hepatoma cells was decreased by 40% and cell number by 15% compared with controls. As cell growth did not resume when the inhibitor was removed from the culture medium, it suggested strongly that ALDHs play a pivotal role in mediating cell death.

Inhibition of cytosolic class 3 aldehyde dehydrogenase by antisense oligonucleotides in rat hepatoma cells.

Aldehyde dehydrogenases (ALDHs) are a superfamily of several isoenzymes widely expressed in bacteria, yeast, plant and animals. Three major classes of ALDHs have been traditionally identified, classes 1, 2 and 3. Both exogenous and endogenous aldehydes, including aldehydes derived from lipid peroxidation, are oxidized by the ALDH superfamily. Several changes in ALDH isoenzyme expression take place in hepatoma cells, in particular cytosolic class 3 ALDH (ALDH3), not expressed in normal hepatocytes, appears and increases with the degree of deviation. It has been demonstrated that cytosolic ALDH3 is important in determining the resistance of tumor cells to antitumor drugs, such as cyclophosphamide. Moreover, hepatoma-associated ALDH3 seems to be important in metabolizing aldehydes derived from lipid peroxidation, and in particular the cytostatic aldehyde 4-hydroxynonenal (4-HNE). We demonstrated previously that restoring endogenous lipid peroxidation in hepatoma cells by enriching them with arachidonic acid causes a decrease of mRNA, protein and enzyme activity of ALDH3 and that this decrease reduces cell growth and/or causes cell death, depending on basal class 3 ALDH activity. To confirm the correlation between inhibition of class 3 ALDH and reduction of cell proliferation, we exposed hepatoma cells to antisense oligonucleotides (ODNs) against ALDH3. In JM2 hepatoma cell line, with high ALDH3 activity, the exposure to antisense ODNs significantly decreases mRNA and enzyme activity (90%). At the same time, cell growth was reduced by about 70%. The results confirm that in hepatoma cells ALDH3 expression is closely related with cell growth, and that its inhibition is important in reducing the proliferation of hepatoma cells overexpressing ALDH3.

Abnormally high TSH in subject with thyrotoxicosis affected by congenital hypothyroidism. Case report.

Inappropriate secretion of TSH (IST) refers to a heterogeneous group of syndromes in which patients show unsuppressed TSH levels in spite of high serum free thyroid hormone concentrations. It has been recognised that IST can be due to both thyroid hormone resistance (RTH) and pituitary TSH-secreting tumours. The former can be generalised (GRTH) or pituitary (PRTH) if the resistance is more severe in the pituitary than in the rest of the tissues. This case report points out the persistence of this patient's TSH resistance to the inhibition of high concentrations of circulating thyroid hormones with clear symptoms of thyrotoxicosis even after many years of replacement therapy; it also suggests that in this case FT4 is the parameter to evaluate the therapy's effectiveness.

Dose-dependent inhibition of cell proliferation induced by lipid peroxidation products in rat hepatoma cells after enrichment with arachidonic acid.

Polyunsaturated fatty acids (PUFA) are important constituents of membrane phospholipids, whose levels are decreased in some tumor cells. This deficiency may cause alterations in signal transduction and an interruption of normal cellular events. The enrichment of tumor cells with PUFA may stimulate or inhibit tumor growth, probably depending on the type of PUFA and the cellular concentration of aldehydes derived from restored lipid peroxidation. We examined the effect of several doses of prooxidant on the growth of hepatoma cells with different aldehyde dehydrogenase activities, enriched with arachidonic acid. Two doses of prooxidant were sufficient to reduce growth of hepatoma cells with low aldehyde dehydrogenase activity, whereas three doses were necessary for those with high enzyme activity. In both cases, lipid peroxidation products blocked the cells in the S phase.

[Increased body mass index increase and insulin resistance parameters in dyslipidemia. Risk and positive correlation in essential obesity]. / Aumento del BMI e parametri di insulino-resistenza e di dislipidemia. Riscontro di correlazioni positive nell'obesità essenziale.

BACKGROUND: Obesity is frequently characterized by hyperinsulinemia with insulin-resistance, tendency to impaired glucose tolerance and by impaired lipidic picture. The relationship existing between some obesity parameters i.e. BMI and weight excess (EP) regarding ideal weight calculated according Lorenz's formula and main parameters both of insulin-resistance and lipidic picture can be interesting. METHODS: 58 obese subject (10 M e 48 F) age 38 +/- 6.2 years (mean +/- DS) with BMI 34.3 +/- 5.3 and EP 33.7 +/- 15; they didn't suffer from hypertension and diabetes; they was subjected to an OGTT and evaluation of glycemia and insulinemia after 0', 60', 90', 120', 150', 180'. Moreover an assessment of lipidic state (cholesterol, triglyceride, HDL, LDL) was carried out. Glycemia and insulinemia with respective areas under curve (AUC glyc. e AUC ins.) and increases (AUCI glyc. e AUCI ins.) were compared with BMI and EP for searching relationship. The same was done for lipidic state. RESULTS: BMI was found positively related with glycemia after 0' (r = 0.3043, p < 0.05) 60' (r = 0.3465, p < 0.05) 120' (r = 0.2895, p < 0.05) with AUC glyc. (r = 0411, p < 0.01) and AUCI glyc. (r = 0.276, p < 0.05), with insulinemia after 0' (r = 0.365, p < 0.01) 60' (r = 0.350, p < 0.01) and with AUC ins. (r = 0.272, p < 0.05). HDL is negatively (r = -0.307, p < 0.05) instead of triglyceride positive related (r = 0.338, p < 0.05) with BMI. EP is positively related with glycemia after 0' (r = 0.376, p < 0.01), 60' (r = 0.362, p < 0.01), 120' (r = 0.290, p < 0.05), with AUC glyc. (r = 0.422, p < 0.01), with insulinemia after 0' (r = 0.512, p < 0.01), 60' (r = 0.473, p < 0.01) with AUC ins. (r = 0.420, p < 0.01) and AUCI ins. (r = 0.354, p < 0.01). Triglyceride was positively related (r = 0.365 p < 0.01) with EP. CONCLUSIONS: These relationships suggest that BMI and EP can be considered as indicators not only of obesity degree, but also of hyperinsulinemia and, to a lesser extent, dyslipidemia severity.