Immunohistochemical Localisation of PDE5 in Rat Lung during Pre- and Postnatal Development

In mammalian lung, at the transition to extrauterine life, NO/cGMP signal transduction system is known to play crucial roles in the regulation of vascular resistance and is supposed to act in angiogenesis. PDE5, which is the most abundant cGMP metabolizing enzyme within the lung, is highly expressed in the perinatal period, but its localisation in the different pulmonary cells is still poorly known. In our research, PDE5 immunohistochemical distribution was investigated in foetal and neonatal rat lung. The highest expression of PDE5 was found in cells randomly located in the stroma; in newborns, in particular, many cells in the intersaccular walls were heavily labelled, while much lower staining levels were shown by smooth myocytes belonging to vessels and airways. On the basis of their immunoreactivity for α-SM actin and/or desmin, most of the heavily PDE5-positive cells were identified as interstitial myofibroblasts and transitional pericytes, while only a few were interpreted as interstitial lipofibroblasts

Preferential association of hepatitis C virus with CD19+ B cells is mediated by complement system

Extrahepatic disease manifestations are common in chronic hepatitis C virus (HCV) infection. The mechanism of HCV-related lymphoproliferative disorders is not fully understood. Recent studies have found that HCV in peripheral blood mononuclear cells (PBMCs) from chronically infected patients is mainly associated with CD19+ B cells. To further elucidate this preferential association of HCV with B cells, we used in vitro cultured virus and uninfected PBMCs from healthy blood donors to investigate the necessary serum components that activate the binding of HCV to B cells. First, we found that the active serum components were present not only in HCV carriers, but also in HCV recovered patients and HCV negative healthy blood donors and that the serum components were heat labile. Second, the preferential binding activity of HCV to B cells could be blocked by anti-complement C3 antibodies. In experiments with complement-depleted serum and purified complement proteins, we demonstrated that complement proteins C1, C2, and C3 were required to activate such binding activity. Complement protein C4 was partially involved in this process. Third, using antibodies against cell surface markers, we showed that the binding complex mainly involved CD21 (complement receptor 2), CD19, CD20, and CD81; CD35 (complement receptor 1) was involved but had lower binding activity. Fourth, both anti-CD21 and anti-CD35 antibodies could block the binding of patient-derived HCV to B cells. Fifth, complement also mediated HCV binding to Raji cells, a cultured B cell line derived from Burkitt´s lymphoma.CONCLUSION:In chronic HCV infection, the preferential association of HCV with B cells is mediated by the complement system, mainly through complement receptor 2 (CD21), in conjunction with the CD19 and CD81 complex. This article is protected by copyright. All rights reserved.Fil: Wang, Richard. National Institutes of Health; Estados UnidosFil: Baré, Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina. National Institutes of Health; Estados UnidosFil: De Giorgi, Valeria. National Institutes of Health; Estados UnidosFil: Matsuura, Kentaro. Nagoya City University Graduate School of Medicine; Japón. National Institutes of Health; Estados UnidosFil: Salam, Kazi Abdus. National Institutes of Health; Estados Unidos. University of Rajshahi; IndiaFil: Grandinetti, Teresa. National Institutes of Health; Estados UnidosFil: Schechterly, Cathy. National Institutes of Health; Estados UnidosFil: Alter, Harvey J.. National Institutes of Health; Estados Unido

Sex differences in the effect of type 2 diabetes on major cardiovascular diseases: results from a population-based study in Italy.

The aim of the study is to assess sex difference in association between type 2 diabetes and incidence of major cardiovascular events, that is, myocardial infarction, stroke, and heart failure, using information retrieved by diabetes register. The inhabitants of Reggio Emilia (Italy) aged 30\u201384 were followed during 2012\u20132014. Incidence rate ratios and 95% confidence intervals were calculated using multivariate Poisson model. The age- and sex-specific event rates were graphed. Subjects with type 2 diabetes had an excess risk compared to their counterparts without diabetes for all the three major cardiovascular events. The excess risk is similar in women and men for stroke (1.8 times) and heart failure (2.7 times), while for myocardial infarction, the excess risk in women is greater than the one observed in men (IRR 2.58, 95% CI 2.22\u20133.00 and IRR 1.78, 95% CI 1.60\u20132.00, resp.; P of interaction < 0.0001). Women had always a lesser risk than men, but in case of myocardial infarction, the women with type 2 diabetes lost part of advantage gained by women free of diabetes (IRR 0.61, 95% CI 0.53\u20130.72 and IRR 0.36, 95% CI 0.33\u20130.39, resp.). In women with type 2 diabetes, the risk of major cardiovascular events is anticipated by 20\u201330 years, while in men it is by 15\u201320

Quality Traits of Medical Cannabis sativa L. Inflorescences and Derived Products Based on Comprehensive Mass-Spectrometry Analytical Investigation

Cannabis sativa L. has been cultivated throughout the world for industrial and medical purposes and is the most controversial plant ever exploited, with considerable discrepancies in the praise and disapproval it receives. Medical Cannabis prescriptions are on the increase in several countries where its therapeutic use is authorised due to its positive role in treating several pathologies even if it represents a multifaceted reality in terms of application. There are at least 550 identified compounds in C. sativa L., including more than 100 phytocannabinoids and 120 terpenes. The chemical complexity of its bioactive constituents highlights the need for standardised and well-defined analytical approaches able to characterise plant chemotype and herbal drug quality as well as to monitor the quality of pharmaceutical cannabis extracts and preparations. This research highlights the potential of using different analytical procedures involving the combination of headspace-solid-phase microextraction (HS-SPME) coupled to GC–MS and accelerated solvent extraction (ASE) coupled to high resolution mass-spectrometry (HPLC-Q Orbitrap®) for the in-depth profiling of quality traits in authorised medical varieties of Cannabis sativa L. flos (Bediol®) and corresponding macerated oil preparations. This approach could add new knowledge to the field of “omic” analytical applications which are fundamental nowadays for Cannabis used for therapeutic remedies

Darolutamide in hormone-sensitive and castration-resistant prostate cancer

Introduction: Important changes in the treatment of prostate cancer have taken place in recent years. Non-metastatic castration-resistant prostate cancer (nmCRPC) has been clinically delineated. In this setting, three drugs have been approved in high-risk disease: apalutamide, enzalutamide and darolutamide.Areas covered:This manuscript aims to profile darolutamide, its clinical development, pharmacologic properties, efficacy and safety. We presented the results of published clinical studies, but we also investigated ongoing ones.Expert opinion: An indirect comparison with the other two aforementioned drugs emerged. While the clinical efficacy is comparable, the toxicity profile is different for darolutamide, resulting in greater tolerance. We must wait for the results of the trials that study darolutamide in hormone-sensitive disease, both in the metastatic phase and in the localized phase. Clinical experience will also be important to determine ever more personalized treatments for patients.</p

How harvest, cleaning and conservation good practices affect the quality of saffron: results of a research conducted in Italy

Saffron (Crocus sativus L.) spice making requires time-spending manual operations: stigmas are separated from the flower picked in bud early in the morning, and once dried they are preserved protected from light. This study verified how the correct pursuing of these good practices affects saffron quality. Few hours of exposure of the flower to the sun determined a significant decrease in the colouring strength (239.66 ± 10.33 versus 255.35 ± 11.87). The correct cleaning of stigmas determined a very significant increase of colouring strength (247.12 ± 13.32 instead of 224.35 ± 14.88) and a significant increase of flavour strength (99.72 ±7.48 against 90.31 ± 6.32, p &lt;0.05). In 24 months, all the samples kept in the dark were still of first category of quality while the ones kept in the light dropped in second category. For all samples there was an increase of aroma strength and a decrease of the flavour and colouring strength, but only the colouring strength loss followed a significantly more sloping trendline. A detectable difference in the content of trans-crocin 4 correlated to the ageing or the way of conservation was not found, nevertheless, it was confirmed that the isomers tend towards a photostationary state where the trans isomer is more present

Identification of murine phosphodiesterase 5A isoforms and their functional characterization in HL-1 cardiac cell line

Phosphodiesterase 5A (PDE5A) specifically degrades the ubiquitous second messenger cGMP and experimental and clinical data highlight its important role in cardiac diseases. To address PDE5A role in cardiac physiology, three splice variants of the PDE5A were cloned for the first time from mouse cDNA library (mPde5a1, mPde5a2 and mPde5a3). The predicted amino acidic sequences of the three murine isoforms are different in the N-terminal regulatory domain. mPDE5A isoforms were transfected in HEK293T cells and they showed high affinity for cGMP and similar sensitivity to sildenafil inhibition. RT-PCR analysis showed that mPde5a1, mPde5a2 and mPde5a3 had differential tissue distribution. In the adult heart, mPde5a1 and mPde5a2 were expressed at different levels whereas mPde5a3 was undetectable. Overexpression of mPDE5As induced an increase of HL-1 number cells which progress into cell cycle. mPDE5A1 and mPDE5A3 overexpression increased the number of polyploid and binucleated cells, mPDE5A3 widened HL-1 areas and modulated hypertrophic markers more efficiently respect to the other mPDE5A isoforms. Moreover, mPDE5A isoforms had differential subcellular localization: mPDE5A1 was mainly localized in the cytoplasm, mPDE5A2 and mPDE5A3 were also nuclear localized. These results demonstrate for the first time the existence of three PDE5A isoforms in mouse and highlight their potential role in the induction of hypertrophy. This article is protected by copyright. All rights reserved

STAT3 can serve as a hit in the process of malignant transformation of primary cells

The transcription factor signal transducer and activator of transcription 3 (STAT3) acts downstream of many pro-oncogenic signals, including cytokines, growth factors and oncogenes, and is accordingly constitutively active in a wide variety of tumors that often become addicted to it. Moreover, STAT3 is a key player in mediating inflammation-driven tumorigenesis, where its aberrant continuous activation is typically triggered by local or systemic production of the pro-inflammatory cytokine IL-6. We recently showed that mouse embryonic fibroblasts (MEFs) derived from STAT3C k/in mice, which express physiological levels of the constitutively active mutant STAT3C, display features of transformed cells such as increased proliferation, resistance to apoptosis and senescence, and aerobic glycolysis. Here, we show that pre-existing constitutively active STAT3 is sufficient to prime primary MEFs for malignant transformation upon spontaneous immortalization. Transformation is strictly STAT3-dependent and correlates with high resistance to apoptosis and enhanced expression of anti-apoptotic/pro-survival genes. Additionally, hypoxia inducible factor (HIF)-1α level is elevated by twofold and contributes to STAT3 oncogenic activity by supporting high rates of aerobic glycolysis. Thus, constitutively active STAT3, an accepted essential factor for tumor growth/progression, can also act as a first hit in multistep carcinogenesis; this ability to predispose cells to malignant transformation may be particularly relevant in the pro-oncogenic niche represented by chronically inflamed tissues

Determinants of inappropriate setting allocation in the care of patients with type 2 diabetes: A population-based study in Reggio Emilia province

The study aims to describe the distribution of patients with type 2 diabetes (T2D) by care plan and to highlight determinants of underuse and overuse of integrated care (IC). This cross-sectional study included all T2D patients resident in Reggio Emilia on 31/12/2015 based on the population-based diabetes registry. Eligibility for IC requires good glycaemic control, no rapid insulin, no kidney failure and no diabetes complications. We calculated the proportion of IC underuse and overuse and adjusted prevalence estimate using multivariate logistic regression. Determinants were age, sex, citizenship, district of residence and time since diagnosis. Of 29,776 patients, 15,364 (51.6%) were in diabetes clinic plan, 9851 (33.1%) in IC plan and 4561 (15.3%) not in any care plan (i.e., in Other group). There were 10,906 (36.6%) patients eligible for IC, of whom 1000 in Other group. When we adjusted for all covariates and restricted the analysis to patients included in care plans, the proportion of those eligible for IC plan but cared for in diabetes clinic plan (i.e. underuse of IC) was 28% (n = 3028/9906; 95%CI 27–29). Similarly, the proportion of those not eligible for IC but cared for in IC plan (i.e. overuse of IC) was 11% (n = 1720/11,896; 95%CI 10–11).The main determinant of both IC underuse and overuse was the district of residence. Foreign status was associated with underuse (37%; 95%CI 33–43), while old age (≥80 years) with both underuse (36%; 95%CI 0.33–0.38) and overuse (23%; 95%CI 22–25). The criterion for suspension of IC plan most frequently found was renal failure, followed by hospitalization for diabetes-related complications. Patients are more often allocated to more specialized settings than not. Healthcare provider-related factors are the main determinants of inappropriate setting allocation

Quality traits of saffron produced in Italy: geographical area effect and good practices

Saffron (Crocus sativus L.) is the most expensive spice in the world and is used in food, cosmetic and dyeing industries. Considering that the production of saffron is increasingly widespread in medium-small Italian farms as well as the scarceness of information and studies regarding the quality of the saffron produced in Italy, the principal aim of this study was to investigate the quality of Italian saffron. Qualitative analysis was conducted in accordance with ISO 3632 1,2:2010-2011 considering 484 samples collected over four years (2015-2018). In particular, moisture content, aroma strength (safranal), colouring strength (crocin) and flavour strength (picrocrocin) were assessed for each sample, and whether spice quality varied according to the geographical area where the spice was produced was also investigated. Qualitative analysis showed that the majority (84-93%) of the samples analysed are of the first quality category, regardless of the year of production. Moisture content and colouring strength are the factors that influence the quality of the spice most. Principal component analysis showed that quality is not influenced by the geographical area where the spice was produced. Finally, some best agricultural practices to obtain a high quality saffron spice are reported