eGLU-Box Mobile:A Smartphone App for Usability Testing by Italian Public Administration Webmasters

Smartphones and tablets now offer consumers unique advantages such as portability and accessibility. Developers are also working with a mobile-first approach, and are prioritizing mobile applications over desktop versions. This study introduces eGLU-box Mobile, an application for performing a drive usability test directly from a smartphone. An experimental study was conducted in which the participants were divided into two groups: an experimental group, which used the new mobile application from a smartphone, and a control group, which used the desktop application from a computer. The participants’ behavior was assessed using explicit (self-report questionnaires) and implicit measures (eye movement data). The results were encouraging, and showed that both the mobile and desktop versions of eGLU-box enabled participants to test the usability with a similar level of UX, despite some minimal (although significant) differences in terms of satisfaction of use

Bioactive Materials for Soft Tissue Repair

Over the past decades, age-related pathologies have increased abreast the aging population worldwide. The increased age of the population indicates that new tools, such as biomaterials/scaffolds for damaged tissues, which display high efficiency, effectively and in a limited period of time, for the regeneration of the body’s tissue are needed. Indeed, scaffolds can be used as templates for three-dimensional tissue growth in order to promote the tissue healing stimulating the body’s own regenerative mechanisms. In tissue engineering, several types of biomaterials are employed, such as bioceramics including calcium phosphates, bioactive glasses, and glass–ceramics. These scaffolds seem to have a high potential as biomaterials in regenerative medicine. In addition, in conjunction with other materials, such as polymers, ceramic scaffolds may be used to manufacture composite scaffolds characterized by high biocompatibility, mechanical efficiency and load-bearing capabilities that render these biomaterials suitable for regenerative medicine applications. Usually, bioceramics have been used to repair hard tissues, such as bone and dental defects. More recently, in the field of soft tissue engineering, this form of scaffold has also shown promising applications. Indeed, soft tissues are continuously exposed to damages, such as burns or mechanical traumas, tumors and degenerative pathology, and, thereby, thousands of people need remedial interventions such as biomaterials-based therapies. It is known that scaffolds can affect the ability to bind, proliferate and differentiate cells similar to those of autologous tissues. Therefore, it is important to investigate the interaction between bioceramics and somatic/stem cells derived from soft tissues in order to promote tissue healing. Biomimetic scaffolds are frequently employed as drug-delivery system using several therapeutic molecules to increase their biological performance, leading to ultimate products with innovative functionalities. This review provides an overview of essential requirements for soft tissue engineering biomaterials. Data on recent progresses of porous bioceramics and composites for tissue repair are also presented

Simultaneous Detection and Viral DNA Load Quantification of Different Human Papillomavirus Types in Clinical Specimens by the High Analytical Droplet Digital PCR Method

Human papillomaviruses (HPVs) are small DNA tumor viruses that mainly infect mucosal epithelia of anogenital and upper respiratory tracts. There has been progressive demand for more analytical assays for HPV DNA quantification. A novel droplet digital PCR (ddPCR) method was developed to simultaneously detect and quantify HPV DNA from different HPV types. DdPCR was initially tested for assay sensitivity, accuracy, specificity as well as intra- and inter-run assay variation employing four recombinant plasmids containing HPV16, HPV18, HPV11, and HPV45 DNAs. The assay was extended to investigate/quantify HPV DNA in Cervical Intraepithelial Neoplasia (CIN, n = 45) specimens and human cell lines (n = 4). DdPCR and qPCR data from clinical samples were compared. The assay showed high accuracy, sensitivity and specificity, with low intra-/inter- run variations, in detecting/quantifying HPV16/18/11/45 DNAs. HPV DNA was detected in 51.1% (23/45) CIN DNA samples by ddPCR, whereas 40% (18/45) CIN tested HPV-positive by qPCR. Five CIN, tested positive by ddPCR, were found to be negative by qPCR. In CIN specimens, the mean HPV DNA loads determined by ddPCR were 3.81 copy/cell (range 0.002–51.02 copy/cell), whereas 8.04 copy/cell (range 0.003–78.73 copy/cell) by qPCR. DdPCR and qPCR concordantly detected HPV DNA in SiHa, CaSki and Hela cells, whereas HaCaT tested HPV-negative. The correlation between HPV DNA loads simultaneously detected by ddPCR/qPCR in CINs/cell lines was good (R2 = 0.9706, p < 0.0001). Our data indicate that ddPCR is a valuable technique in quantifying HPV DNA load in CIN specimens and human cell lines, thereby improving clinical applications, such as patient management after primary diagnosis of HPV-related lesions with HPV-type specific assays

MicroRNAs Modulate Signaling Pathways in Osteogenic Differentiation of Mesenchymal Stem Cells

Mesenchymal stem cells (MSCs) have been identified in many adult tissues and they have been largely studied in the last years, especially in view of their potential use for treating diseases, damaged tissues and organs. MSCs are capable of self-replication and differentiation into osteo-blasts and are considered an important source of cells in tissue engineering for bone regeneration. Several epigenetic factors are believed to play a role in the osteogenic differentiation of MSCs, in-cluding microRNAs (miRNAs). MiRNAs are small, single-stranded, non-coding RNAs of approxi-mately 22 nucleotides able to regulate cell proliferation, differentiation and apoptosis by binding the 3′ untranslated region (3′-UTR) of target mRNAs, which can be subsequently degraded or trans-lationally silenced. MiRNAs control gene expression in osteogenic differentiation by regulating two crucial signaling cascades in osteogenesis: the transforming growth factor-beta (TGF-β)/bone mor-phogenic protein (BMP) and the Wingless/Int-1(Wnt)/β-catenin signaling pathways. This review provides an overview of the miRNAs involved in osteogenic differentiation and how these miRNAs could regulate the expression of target genes

Chronic lymphocytic leukemia tested positive for the oncogenic Merkel Cell Polyomavirus, MCC-350 strain

Merkel cell carcinoma (MCC) have been found to be associated with the oncogenic Merkel cell polyomavirus (MCPyV) [1]. However, MCPyV sequences have been detected at low prevalence in buffy coats [2] and sera of blood donors [3]. Chronic lymphocytic leukemia (CLL) were also found associated with MCPyV by some investigators, whereas other studies did not confirm this result. In our investigation, DNA sequences belonging to MCPyV were identified with a different prevalence in sera from CLL patients and blood donors. MCPyV sequences in sera of CLL patients and blood donors had a prevalence of 7% (16/224) and 5% (14/284), respectively. Specifically, MCPyV DNA sequences, coding for the viral oncoprotein large T antigen (LT), analyzed by the droplet digital polymerase chain reaction (ddPCR) method and DNA sequencing, showed the circulation of two different MCPyV strains, MCC350 and MKL-1. Indeed, DNA sequencing performed in MCPyVpositive sera indicated that MCPyV LT sequences belong to the ubiquitous MKL-1 [1-2] and oncogenic MCC350 strains. Interestingly, the more oncogenic MCC-350 strain was present at higher prevalence, 81% (13/16), in CLL samples, while the prevalence of MCC-350 was only 21% (3/14) in sera of blood donors (P<0.05). It is worth recalling that MCPyV MCC350 was the strain originally identified in MCC, a rare skin cancer. In our study, this oncogenic strain is found to be more prevalent in CLL patients, whereas the ubiquitous MKL-1 is more prevalent in blood donors. These data suggest that the MCC 350 strain could be responsible of the CLL onset in a fraction of these patients, whereas the MKL-1 strain seems to be the MCPyV circulating in normal subjects

METHYLATION PROFILE OF IRF6 AND RARB GENE PROMOTERS IN NORMAL VULVAR TISSUES AND VULVAR CARCINOMAS

Interferon regulatory factor 6 (IRF6) and retinoic acid receptor beta (RARB) play an important role in regulating cell proliferation and differentiation of the epithelia. IRF6 and RARB by modulating p63 and c-jun, respectively, arrest cell proliferation thus inducing differentiation. The promoter hypermethylation of tumor suppressor genes may favour the onset of cancers. In this study, IRF6 and RARB promoter methylation profiles were investigated in normal vulvar (NV, n=20) and pathological vulvar tissues from cancer-free lichen sclerosus (cfLS, n=20), cancer-associated lichen sclerosus (caLS, n=20), vulvar intraepithelial neoplasia (VIN, n=6, only IRF6) and vulvar cancer (VC, n=20) specimens. Methylation analyses were performed with the bisulphite-DNA treatment and PCR amplifications/sequencing of IRF6 and RARB promoters. IRF6 and RARB gene expressions, together with p63 and c-jun genes were analysed by qRT-PCR. IRF6 gene promoter was found to be hypermethylated in 10% cfLS, 20% VIN, 45% caLS and 80% VC (p<0.01, caLS and VC versus NV). IRF6 expression decreased 2.2-, 2.9-, 4.5- and 6.6-fold from cfLS, VIN, caLS to VC, respectively, whereas p63 was overexpressed in all specimens compared to NV (p<0.05). RARB gene promoter tested hypermethylated in 50% caLS, 55% cfLS and 90% VC (p<0.01, versus NV). Unlike IRF6, RARB was significantly down-expressed, 4.8-fold, only in VC (p<0.01, versus NV). Consistently, c-jun expression was 2.6-fold up-expressed in VC (p<0.01, versus NV). Interestingly, 2/18 (11.1%) VC, showing full methylation of RARβ gene promoter, were from females with tumor recurrences. IRF6 and RARB expressions are hampered by promoter hypermethylation in vulvar diseases and vulvar cancer. While IRF6 promoter hypermethylation occurs in a stepwise manner from vulvar LS to cancer, RARB promoter hypermethylation was found to be mainly associated to vulvar cancer. IRF6 and RARB dysregulations may play a role in caLS development and progression, respectively

HYPERMETHYLATION-INDUCED INACTIVATION OF IRF6 AND RARβ GENES AS A POTENTIAL PROGNOSTIC BIOMARKER IN VULVAR SQUAMOUS CELL CARCINOMA

Objectives: Vulvar squamous cell carcinoma (VSCC) represents 5% of gynecological malignancies. About 80% of VSCCs arises from inflammatory diseases, such as lichen sclerosus (LS). The molecular alterations involved in onset/progression of LS-associated VSCC are unknown. Interferon regulatory factor 6 (IRF6) and retinoic acid receptor β (RARβ) tumor-suppressor genes have been found to be downregulated by promoter methylation in different carcinomas.1,2 In this study, we aimed to evaluate the involvement of the IRF6 and RARβ promoter methylation in the onset of VSCC from LS. Methods: IRF6 and RARβ mRNA expression and promoter methylation profiles were investigated by quantitative PCR (qPCR) and sequencing of PCR-amplified bisulfite-treated DNA, respectively, in VSCC (n=20) and the adjacent LS (n=20), vulvar intraepithelial neoplasia (VIN; n=5, only for IRF6), cancer-free LS (cfLS, n=20) and normal skin (NS, n=20). IRF6 and RARβ pathway-related genes p63 and c-Jun mRNAs were also investigated by qPCR.3,4 Results: IRF6 expression decreased 2.2-, 2.9-, 4.5- and 6.6-fold from cfLS, VIN, LS to VC, respectively, whereas p63 was overexpressed in all specimens compared to NS (p<0.05). IRF6 promoter was hypermethylated in 9/20 (45%) LS, 1/5 (20%) VIN, 16/20 (80%) VSCC, 2/20 (10%) cfLS, and 0 NS.3 Unlike IRF6, RARB was significantly down-expressed, 4.8-fold, only in VC (p<0.01, versus NS). Consistently, c-jun expression was 2.6-fold up-expressed in VC (p<0.01, versus NV). RARβ gene promoter was hypermethylated in 18/20 (90%) VSCC, 11/20 (55%) cfLS, 10/20 (50%) LS and 5/20 (25%) NS. Interestingly, 2/18 (11.1%) VSCC from females with tumor recurrences showed full methylation of RARβ gene promoter.4 Conclusions: IRF6 and RARB gene expressions are hampered by promoter hypermethylation in vulvar diseases and vulvar cancer. While IRF6 promoter hypermethylation occurs in a stepwise manner from vulvar LS to cancer, RARB promoter hypermethylation was found to be mainly associated to vulvar cancer. IRF6 and RARB dysregulations may play a role in LS development and progression, respectively. Therefore, IRF6 and RARβ promoter hypermethylations may be biomarkers of cancer-risk in LS patients, and prognostic biomarkers of cancer progression in patients affected by LS-associated-VSCC.3,4 References: 1.Ivanova T et al. BMC Cancer.2:4,2002. 2.Botti E et al. PNAS.108:13710-15,2011. 3.Rotondo JC et al. JAMA Dermatol.152:928-33,2016. 4.Rotondo JC et al. JAMA Dermatol.154:1-5,2018

Mother-to-child transmission of oncogenic polyomaviruses BKPyV, JCPyV and SV40

Objectives: Polyomavirus (PyV) infections have been associated with different diseases. BK (BKPyV), JC (JCPyV) and simian virus 40 (SV40) are the three main PyVs whose primary infection occurs early in life. Their vertical transmission was investigated in this study. Methods: PyV sequences were analyzed by the digital droplet PCR in blood, serum, placenta, amniotic fluid, vaginal smear from two independent cohorts of pregnant females and umbilical cord blood (UCB) samples. IgG antibodies against the three PyVs were investigated by indirect E.L.I.S.As with viral mimotopes. Results: DNAs from blood, vaginal smear and placenta tested BKPyV-, JCPyV- and SV40- positive with a distinct prevalence, while amniotic fluids were all PyVs-negative. A prevalence of 3%, 7%, and 3% for BKPyV, JCPyV and SV40 DNA sequences, respectively, was obtained in UCBs. Serum IgG antibodies from pregnant females reached an overall prevalence of 62%, 42% and 17% for BKPyV, JCPyV and SV40, respectively. Sera from newborns (UCB) tested IgGpositive with a prevalence of 10% for BKPyV/JCPyV and 3 % for SV40. Conclusions: In this investigation, PyV vertical transmission was revealed by detecting PyV DNA sequences and IgG antibodies in samples from females and their offspring suggesting a potential risk of diseases in newborns

The role of microRNAs in the osteogenic and chondrogenic differentiation of mesenchymal stem cells and bone pathologies

Mesenchymal stem cells (MSCs) have been identified in many adult tissues. MSCs can regenerate through cell division or differentiate into adipocytes, osteoblasts and chondrocytes. As a result, MSCs have become an important source of cells in tissue engineering and regenerative medicine for bone tissue and cartilage. Several epigenetic factors are believed to play a role in MSCs differentiation. Among these, microRNA (miRNA) regulation is involved in the fine modulation of gene expression during osteogenic/chondrogenic differentiation. It has been reported that miRNAs are involved in bone homeostasis by modulating osteoblast gene expression. In addition, countless evidence has demonstrated that miRNAs dysregulation is involved in the development of osteoporosis and bone fractures. The deregulation of miRNAs expression has also been associated with several malignancies including bone cancer. In this context, bone-associated circulating miRNAs may be useful biomarkers for determining the predisposition, onset and development of osteoporosis, as well as in clinical applications to improve the diagnosis, follow-up and treatment of cancer and metastases. Overall, this review will provide an overview of how miRNAs activities participate in osteogenic/chondrogenic differentiation, while addressing the role of miRNA regulatory effects on target genes. Finally, the role of miRNAs in pathologies and therapies will be presented