Publications by authors named "Cristina Calvi"

28 Publications

  • Page 1 of 1

Macrophage Receptor With Collagenous Structure Polymorphism and Recurrent Respiratory Infections and Wheezing During Infancy: A 5-Years Follow-Up Study.

Front Pediatr 2021 27;9:666423. Epub 2021 Jul 27.

Department of Public Health and Pediatric Sciences, Paediatric Laboratory, Medical School, University of Turin, Turin, Italy.

Recurrent wheezing is a common clinical manifestation in childhood, and respiratory syncytial virus infection is a well-known risk factor. However, the genetic background favoring the development of recurrent wheezing is not fully understood. A possible role of macrophage receptor with collagenous gene (MARCO) polymorphism has been recently proposed. To investigate a correlation between MARCO rs1318645 polymorphisms and susceptibility to recurrent wheezing during childhood. We prospectively recruited 116 infants, of which 58 with respiratory syncytial virus bronchiolitis and 58 controls hospitalized at Regina Margherita Children's Hospital, Turin, Italy, between November 2014 and April 2015. All subjects were investigated for MARCO rs1318645 polymorphisms in the first period of life. Genotyping of rs1318645 was carried out by TaqMan mismatch amplification mutation assay real-time polymerase chain reaction procedure. Subjects were then enrolled in a 5-year follow-up study to monitor the occurrence of wheezing and respiratory infections. The analysis of MARCO rs1318645 of allelic frequencies shows an increasingly significant risk to develop recurrent infection ( = 0.00065) and recurrent wheezing ( = 0.000084) with a wild-type C allele compared with a G allele. No correlation was found between wheezing and past respiratory syncytial virus infection ( = 0.057) and for a history of atopy in the family ( = 0.859). Our finding showed that subjects with C allelic MARCO rs1318645 polymorphism are at higher risk for recurrent infection and wheezing episodes during the first 5 years of life. Future studies of genetic associations should also consider other types of polymorphisms.
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http://dx.doi.org/10.3389/fped.2021.666423DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8353117PMC
July 2021

COVID-19 in Children: Expressions of Type I/II/III Interferons, TRIM28, SETDB1, and Endogenous Retroviruses in Mild and Severe Cases.

Int J Mol Sci 2021 Jul 13;22(14). Epub 2021 Jul 13.

Department of Pediatric Sciences and Public Health, University of Turin, Piazza Polonia 94, 10126 Turin, Italy.

Children with the new coronavirus disease 2019 (COVID-19) have milder symptoms and a better prognosis than adult patients. Several investigations assessed type I, II, and III interferon (IFN) signatures in SARS-CoV-2 infected adults, however no data are available for pediatric patients. TRIM28 and SETDB1 regulate the transcription of multiple genes involved in the immune response as well as of human endogenous retroviruses (HERVs). Exogenous viral infections can trigger the activation of HERVs, which in turn can induce inflammatory and immune reactions. Despite the potential cross-talks between SARS-CoV-2 infection and TRIM28, SETDB1, and HERVs, information on their expressions in COVID-19 patients is lacking. We assessed, through a PCR real time Taqman amplification assay, the transcription levels of six IFN-I stimulated genes, IFN-II and three of its sensitive genes, three IFN-lIIs, as well as of TRIM28, SETDB1, pol genes of HERV-H, -K, and -W families, and of env genes of Syncytin (SYN)1, SYN2, and multiple sclerosis-associated retrovirus (MRSV) in peripheral blood from COVID-19 children and in control uninfected subjects. Higher expression levels of IFN-I and IFN-II inducible genes were observed in 36 COVID-19 children with mild or moderate disease as compared to uninfected controls, whereas their concentrations decreased in 17 children with severe disease and in 11 with multisystem inflammatory syndrome (MIS-C). Similar findings were found for the expression of TRIM-28, SETDB1, and every HERV gene. Positive correlations emerged between the transcriptional levels of type I and II IFNs, TRIM28, SETDB1, and HERVs in COVID-19 patients. IFN-III expressions were comparable in each group of subjects. This preserved induction of IFN-λs could contribute to the better control of the infection in children as compared to adults, in whom IFN-III deficiency has been reported. The upregulation of IFN-I, IFN-II, TRIM28, SETDB1, and HERVs in children with mild symptoms, their declines in severe cases or with MIS-C, and the positive correlations of their transcription in SARS-CoV-2-infected children suggest that they may play important roles in conditioning the evolution of the infection.
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http://dx.doi.org/10.3390/ijms22147481DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8303145PMC
July 2021

Comparison of methods for isolating fungal DNA.

Pract Lab Med 2021 May 17;25:e00221. Epub 2021 Apr 17.

Department of Public Health and Pediatrics, University of Turin, School of Medicine, Turin, Italy.

Objectives: The main aim of this work was to compare the methods of DNA isolation in the moulds of genus with special regard to the amount and purity of the DNA acquired. The acquired DNA was then amplified by specific real-time PCR.

Design: Five DNA extraction procedures were carried out in a Class 2 Biosafety cabinet in a dedicated room with suitable biosafety precautions and appropriate biowaste disposal methods. A total of 6 clinical strains were used.

Results: From the viewpoint of concentration and purity, methods A shown abundant amount of fungal DNA whereas methods E report a pure fungal DNA with R260/280 of 1.7 near the optimal 1.8. The DNA quantity reach statistically difference at ANOVA test with p value 0.0005.

Conclusion: Overall, the E method was the most efficient method in the extraction of DNA from fungal cultures compared to the other methods considering time, cost, technical expertise, and instrumentation. Use of this assay will allow researchers to obtain DNA from fungi quickly for use in molecular assays.
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http://dx.doi.org/10.1016/j.plabm.2021.e00221DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8145739PMC
May 2021

TaqMan real time PCR for the Detection of the Gilbert's Syndrome Markers UGT1A1*28; UGT1A1*36 and UGT1A1*37.

Mol Biol Rep 2021 May 4;48(5):4953-4959. Epub 2021 Jun 4.

Department of Public Health and Pediatrics, School of Medicine, University of Turin, Piazza Polonia 94, 10126, Turin, Italy.

Gilbert's syndrome is characterized by mild unconjugated hyperbilirubinemia. The key of this disease is a diminished activity of UDP-glucuronosyltransferase 1A1 (UGT1A1). TA insertion into the TATA box promoter region of the UGT1A1 gene on chromosome 2 is the genetic basis of Gilbert's syndrome (UGT1A1*28). An extra TA insert leads to eight (TA)8 repeats (UGT1A1*37) resulting in a further reduction of glucuronidation activity. A variant lacking one TA repeat (TA)5 (UGT1A1*36) has been identified. (TA)8 repeats (UGT1A1*37) and (TA)5 (UGT1A1*36) have been detected in Africans (frequency up to 0.07 and 0.08 respectively). We present a real time PCR method for genotyping the UGT1A1 (TA)n polymorphism (UGT1A1*28, UGT1A1*36, UGT1A1*37) using Taqman PCR on 7500 and cfx96 Real-Time PCR System. We present a real time PCR method for genotyping the UGT1A1 (TA)n polymorphism (UGT1A1*28, UGT1A1*36, UGT1A1*37) using Taqman PCR. About clinical validation, all 53 samples collected from patients referred for suspected Gilbert's syndrome were analyzed. We found 21 on the 53 patients (39.6%) were homozygotes (UGT1A1-TATA (TA)6) and referred as wild-type, 13 on the 53 patients (24.5%) were homozygotes (UGT1A1-TATA (TA)7) and referred as mutated, 1 on the 53 patients (1.9%) were homozygotes (UGT1A1-TATA (TA)8) and referred as mutated, 1 on the 53 patients (1.9%) were heterozygotes (UGT1A1-TATA (TA)7/8) and referred as mutated, 1 on the 53 patients (1.9%) were heterozygotes (UGT1A1-TATA (TA)5/6) and referred as mutated, and 16 on the 53 patients (30.2%) were heterozygotes (UGT1A1-TATA (TA)6/7). None were homozygotes UGT1A1-TATA (TA)5, homozygotes UGT1A1-TATA (TA)8, or heterozygotes with (TA)5 or (TA)8 alleles. The newly described technique represents a valid alternative method to sequencing, mainly due to its rapidity, easiness, and minor costs.
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http://dx.doi.org/10.1007/s11033-021-06454-2DOI Listing
May 2021

Bufavirus, Cosavirus, and Salivirus in Diarrheal Italian Infants.

Intervirology 2021 30;64(3):165-168. Epub 2021 Mar 30.

Department of Public Health and Pediatric Sciences, Pediatrics Laboratory, University of Turin, Medical School, Turin, Italy.

Three newly discovered viruses have been recently described in diarrheal patients: Cosavirus (CosV) and Salivirus (SalV), 2 picornaviruses, and bufavirus (BuV), a parvovirus. The detection rate and the role of these viruses remain to be established in acute gastroenteritis (AGE) in diarrheal Italian infants. From November 2016 to November 2017, stool samples were collected from 160 children <5 years old suffering from AGE and attending the Children's Hospital in Turin, Italy. During the study period, 1 (0.5%) sample was positive for 1 of the 3 investigated viruses: 0 (0%) CosV, 1 (0.5%) SalV, and 0 (0%) BuV, whereas 42 (26.0%) children were infected with rotavirus and 2 (1%) with adenovirus. No mixed infections involving the 3 viruses were found. Although these viruses are suspected to be responsible for AGE in children, our data showed that this association was uncertain. Therefore, further studies with large cohorts of healthy and diarrheal children will be needed to evaluate their clinical role in AGE.
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http://dx.doi.org/10.1159/000514384DOI Listing
March 2021

Overexpression of endogenous retroviruses in children with celiac disease.

Eur J Pediatr 2021 Aug 26;180(8):2429-2434. Epub 2021 Mar 26.

Department of Public Health and Pediatric Sciences, University of Turin, Piazza Polonia, 96 10126, Turin, Italy.

Human endogenous retroviruses (HERVs) represent 8% of our genome. Although no longer infectious, they can regulate transcription of adjacent cellular genes, produce retroviral RNAs, and encode viral proteins that can modulate both innate and adaptive immune responses. Based on this, HERVs have been studied and proposed as contributing factors in various autoimmune disorders. Celiac disease (CD) is considered an autoimmune disease, but HERV expression has not been studied in celiac patients. The aim of this study is to assess the transcription levels of pol genes of HERV-H, -K, and -W and of their TRIM28 repressor in WBCs from celiac children and age-matched control subjects. A PCR real-time TaqMan amplification assay was used to evaluate HERV and TRIM28 transcripts with normalization of the results to glyceraldehyde-3-phosphate dehydrogenase. The RNA levels of pol genes of the three HERV families were significantly higher in WBCs from 38 celiac patients than from 51 control subjects. TRIM28 transcription was comparable between the two study populations.Conclusion: Present results show, for the first time, that pol genes of HERV-H, -K, and -W are overexpressed in patients with CD. Given their proinflammatory and autoimmune properties, this suggests that HERVs may contribute to the development of CD in susceptible individuals. What is Known: • Based on this, HERVs have been studied and proposed as contributing factors in various autoimmune disorders. What is New: • Present results show, for the first time, that pol genes of HERV-H, -K, and -W are overexpressed in patients with CD.
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http://dx.doi.org/10.1007/s00431-021-04050-xDOI Listing
August 2021

Lack of detection of HPyV12 DNA using real-time PCR in Italian infants with diarrhea.

Minerva Pediatr 2020 Jun 5. Epub 2020 Jun 5.

Department of Public Health and Pediatric Sciences, University of Turin Medical School, Turin, Italy.

Background: HPyV12 was found in organs of the digestive tract, in particular the liver but also in colon, rectum and faeces. Until now, the prevalence of HPyV12 is not well.characterized.

Methods: In this study, we investigate the presence of this novel polyomavirus DNA in stool specimens collected from under-five-year-old children with gastroenteritis compared to healthy infants. A total of 190 fecal specimens previously screened for Rotavirus (RV) and Adenovirus (ADV) and 80 fecal samples from healthy infants, were tested for HPyV12 DNA using a home-made real time PCR. All fecal specimens were tested for the presence of HPyV12 with specific primers and probes.

Results: None of 190 (0%) episodes of acute gastroenteritis was associated with HPyV12. We did not detect HPyV12 DNA in any of 80 control subjects, as well.

Conclusions: Our study represents a pilot study aiming to clarify the current epidemiological pattern in pediatric italian patients regarding the novel and rare HPyV12. On the basis of our negative data and the recently observations reported in literature, doubts remain on human tropism of the HPyV12 and epidemiology: these issues need further investigations.
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http://dx.doi.org/10.23736/S0026-4946.20.05738-2DOI Listing
June 2020

Chronic HCV Infection Is Associated with Overexpression of Human Endogenous Retroviruses that Persists after Drug-Induced Viral Clearance.

Int J Mol Sci 2020 Jun 1;21(11). Epub 2020 Jun 1.

Department of Pediatric Sciences and Public Health, University of Turin, Piazza Polonia 94, 10126 Turin, Italy.

Chronic hepatitis C virus (HCV) infection is associated with several hepatic and extrahepatic complications, including cancers and autoimmune disorders, whose frequency is reduced but not abolished after drug-induced viral clearance. The causes of these complications and of their persistence are ill-defined. Human endogenous retroviruses (HERVs) are remnants of ancestral infections and constitute 8% of the human genome. Most HERV elements are inactive, but some are transcribed. HERV overexpression is associated with many cancers and autoimmune diseases with a putative pathogenetic role. Several viral infections trigger HERV activation, but there are no studies on HCV-infected subjects. We assessed, through a PCR real-time amplification assay, the transcription levels of the pol genes of HERV-H, -K, and -W, and of their repressor TRIM28 in white blood cells (WBCs) of vertically infected children, both before and after therapy with direct-acting antivirals (DAAs). The results documented significantly higher expressions of HERV-H-pol and HERV-K-pol, not of HERV-W-pol, in HCV-infected subjects as compared to age-matched controls. HERV RNA levels remained unchanged after DAA-driven viral clearance. No significant variations in transcription levels of TRIM28 were observed in infected subjects. Our findings demonstrate HERV-H-pol and HERV-K-pol overexpression in subjects with chronic HCV infection, without variations after a positive response to DAAs; this might justify their predisposition to cancers and autoimmune disorders that persist after a DAA-induced resolution of viremia.
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http://dx.doi.org/10.3390/ijms21113980DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7313012PMC
June 2020

Modulation of human endogenous retroviruses -H, -W and -K transcription by microbes.

Microbes Infect 2020 09 6;22(8):366-370. Epub 2020 Feb 6.

Department of Pediatrics, Azienda Ospedaliera Universitaria Città della Salute e della Scienza di Torino, Turin, Italy. Electronic address:

The human endogenous retroviruses (HERVs) are endogenous retroviruses that are inserted into the germ cell DNA of humans over 30 million years ago. Using real-time RT-PCR we describe HERV modulation by commensal microbes in the human gut. Infants, exclusively or predominant breast milk feeding, less than 12 weeks of age, during bacteria gut colonization, were assessed for eligibility. Our data demonstrate that the colonization with commensal microbes, in particular, Bifidobacterium spp., of the gut causes modulation of HERVs.
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http://dx.doi.org/10.1016/j.micinf.2020.01.006DOI Listing
September 2020

Mir-155 expression is downregulated in hematopoietic stem cell transplantation patients with Epstein-Barr virus infection.

Minerva Pediatr 2019 Dec 11. Epub 2019 Dec 11.

Department of Public Health and Pediatric Sciences, University of Turin, Medical School, Turin, Italy -

Background: MicroRNAs (miRNAs) are a class of short length double strand genome-encoded RNAs that are produced to repress post- transcriptionally the expression of cellular mRNAs. 2578 unique mature miRNAs are currently annotated in the human genome and participate in the regulation of multiple events, such as cellular proliferation or apoptosis. The over-expression of miR-155 of cellular origin might play a key role in the life cycle of EBV.

Objectives: In this study 24 paediatric patients undergoing HSCT seropositive and seronegative to EBV were enrolled. Thirty-one peripheral blood samples were collected from these patients. The mir-155 expression profile has been evaluated by a stem-loop Real Time PCR in all these conditions.

Study Designs: Of 24 patients, 4 were seronegative to EBV and EBV negative to PCR (Group I), 10 were seropositive to EBV and EBV negative to PCR (Group II) and 10 were seropositive to EBV and EBV positive to PCR (Group III).

Results: Based on relative quantification, the mir-155 expression was compared among the groups. The comparison between HSCT patients without EBV infection seronegative to EBV (Group I) showed higher levels of mir-155 expression than patients seropositive to EBV (p=0.1419). The mir-155 expression levels in seronegative to EBV were not significantly different compared with the patients seropositive to EBV ( p=0.6504). The mir-155 expression levels in seropositive to EBV without and with EBV infection (positive viral load), were not significantly (p=0.7667). Also when we evaluated the mir-155 expression levels comparing all EBV negative patients with an active EBV infection, we did not observe a statistical significant difference ( p=0.9782).

Conclusions: Our result is controversial as the results obtained by Gao et al. showed a higher production of mir-155 levels during EBV infection.
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http://dx.doi.org/10.23736/S0026-4946.19.05420-3DOI Listing
December 2019

Quantification of fecal adenovirus viral load and correlation with Vesikari score in children with acute gastroenteritis.

Minerva Pediatr 2019 Oct 11. Epub 2019 Oct 11.

Pediatric Laboratory, Department of Public Health and Pediatric Sciences, University of Turin Medical School, Turin, Italy.

Background: Human adenoviruses (HAdVs) are an important cause of acute respiratory tract infections, conjunctivitis, hemorrhagic cystitis, and gastroenteritis. In addition to enteric serotypes 40 and 41, some serotypes belonging to subgroups A, B, and C have also been implicated to be etiological agents of gastroenteritis among infants and young children.The Vesikari Scoring System (VSS) is the severity scale that was originally developed to evaluate the effectiveness and efficacy of rotavirus vaccines on 20 points. The objective of this study was to evaluate and compare the diagnostic value of the VSS with HAdVs genome quantification in fecal samples collected from hospitalized children with acute gastroenteritis.

Methods: A total of 137 fecal specimens (69 male and 68 female) were tested for HAdVs. The samples were collected from under-five-year-old children with acute gastroenteritis in pediatric Hospital Regina Margherita of Turin in Italy.

Results: A total of 69 out of 137 (50.3%) samples were associated with HAdV genomic detection with a mean viral load of 1.08x1011 ± 9.02x1011 genomes/mg fecal specimens. The samples were grouped on the basis of Mild VSS and Moderate VSS and the HAdV viral load was calculated in the two groups. No statistical differences were observed between two groups (p= 0.6123 calculated by Mann-Whitney test).

Conclusions: Our results did not show a difference in mean viral load between the group with mild VVS and moderate VVS.
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http://dx.doi.org/10.23736/S0026-4946.19.05510-5DOI Listing
October 2019

DSM 17938 Probiotics May Increase CC-Chemokine Receptor 7 Expression in Infants Treated With for Colic.

Front Pediatr 2019 16;7:292. Epub 2019 Jul 16.

Department of Public Health and Paediatric Sciences, Scuola di Medicina, Università degli Studi di Torino, Turin, Italy.

Studies have shown that ri probiotics can affect cells that play a key role in the immune system. This Italian study investigated how DSM 17938 influenced CC-chemokine receptor 7 (CCR7) and interleukin 10 (IL-10) in breastfed colicky infants. Our University hospital in Turin recruited 50 healthy outpatients, at a median age of approximately 1 month, from September 2017 to August 2018. They were randomized to daily DSM17938 (1 × 10 cfu) or a placebo for 28 days from recruitment. We collected peripheral blood and evaluated the expression of CCR7 messenger ribonucleic acid using the real-time TaqMan reverse transcription polymerase chain reaction method at baseline and after the study period. We found increased expression of CC-chemokine receptor 7 in infants treated with the probiotic, but not the controls ( < 0.0026). No differences were observed for interleukin 10 after the study period in either group. At baseline, daily crying time was comparable in the probiotic and control groups: 341 (25) vs. 337 (29) min., respectively ( = 0.450). After 28 days, daily mean crying time decrease statistically in the probiotic group: 78 (23) vs. 232 (31), respectively ( < 0.001). The increase in CC-chemokine receptor 7 might have been a response to probiotic treatment. As a relatively small sample was used to conduct this study, our research needs to be replicated in different settings, and over time, to produce comparable findings.
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http://dx.doi.org/10.3389/fped.2019.00292DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6646728PMC
July 2019

Peritoneal mesothelioma and asbestos exposure: a population-based case-control study in Lombardy, Italy.

Occup Environ Med 2019 08 8;76(8):545-553. Epub 2019 Jul 8.

Occupational Health Unit, Fondazione IRCCS Ca' Granda - Ospedale Maggiore Policlinico, Milan, Italy.

Objectives: Asbestos is the main risk factor for peritoneal mesothelioma (PeM). However, due to its rarity, PeM has rarely been investigated in community-based studies. We examined the association between asbestos exposure and PeM risk in a general population in Lombardy, Italy.

Methods: From the regional mesothelioma registry, we selected PeM cases diagnosed in 2000-2015. Population controls (matched by area, gender and age) came from two case-control studies in Lombardy on lung cancer (2002-2004) and pleural mesothelioma (2014). Assessment of exposure to asbestos was performed through a quantitative job-exposure matrix (SYN-JEM) and expert evaluation based on a standardised questionnaire. We calculated period-specific and gender-specific OR and 90% CI using conditional logistic regression adjusted for age, province of residence and education.

Results: We selected 68 cases and 2116 controls (2000-2007) and 159 cases and 205 controls (2008-2015). The ORs for ever asbestos exposure (expert-based, 2008-2015 only) were 5.78 (90% CI 3.03 to 11.0) in men and 8.00 (2.56 to 25.0) in women; the ORs for definite occupational exposure were 12.3 (5.62 to 26.7) in men and 14.3 (3.16 to 65.0) in women. The ORs for ever versus never occupational asbestos exposure based on SYN-JEM (both periods) were 2.05 (90% CI 1.39 to 3.01) in men and 1.62 (0.79 to 3.27) in women. In men, clear positive associations were found for duration, cumulative exposure (OR 1.33 (1.19 to 1.48) per fibres/mL-years) and latency.

Conclusions: Using two different methods of exposure assessment we provided evidence of a clear association between asbestos exposure and PeM risk in the general population.
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http://dx.doi.org/10.1136/oemed-2019-105826DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6703122PMC
August 2019

Comparison of Quantitative Analysis of Methylated Alleles Real-Time PCR and Methylation-Specific MLPA for Molecular Diagnosis of Beckwith-Wiedemann Syndrome.

Pathobiology 2019 25;86(4):217-224. Epub 2019 Jun 25.

Department of Public Health and Pediatric Sciences, University of Turin Medical School, Turin, Italy.

Background/aims: Beckwith-Wiedemann syndrome (BWS) is a congenital overgrowth disorder predisposing to tumorigenesis caused by abnormal expression or function of imprinted genes of the chromosome 11p15.5 imprinting gene cluster. This real-time PCR-based assay determines the methylation status of a selected CpG island and has been proposed for use in high-throughput methylation analysis.

Methods: Here, we use quantitative analysis of methylated alleles (QAMA) for the detection of methylation status of the KCNQ10T1 gene, in a region immediately upstream of the transcription initiation site, and the CTCF binding site 6, located approximately 2 kb upstream of the SmaI site currently used for clinical laboratory testing. We assayed a series of controls and patients diagnosed with BWS at two different loci at 11p15.5 to assess the diagnostic yield of QAMA PCR for clinical laboratory testing.

Results: These results compare favorably with methylation-specific multiple ligation probe amplification (MS-MLPA) analysis at both differentially methylated region (DMR)1 and DMR2. There are several advantages of the QAMA PCR over MS-MLPA. The QAMA PCR is less labor-intensive and therefore more cost-effective and does not require dedicated analysis software. A second advantage is that the assay is amenable to high-throughput analysis.

Conclusions: The small sample size reflects the rare nature of this epigenetic disorder, and the range of ages was quite wide, as was the degree of disease severity. Therefore, further validation with larger cohorts is warranted.
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http://dx.doi.org/10.1159/000500627DOI Listing
December 2019

Human endogenous retroviruses HERV-H, HERV-W and HERV-K in preeclampsia.

Minerva Ginecol 2019 Jun 12;71(3):260-262. Epub 2019 Feb 12.

Citoimmunodiagnostics Laboratory, Department of Public Health and Pediatric Sciences, University of Turin, Medical School, Turin, Italy.

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http://dx.doi.org/10.23736/S0026-4784.19.04321-1DOI Listing
June 2019

Human bocavirus in children with acute gastroenteritis in Piedmont, Italy.

Minerva Pediatr 2019 Jan 2. Epub 2019 Jan 2.

Citoimmunodiagnostics Laboratory, Department of Public Health and Pediatric Sciences, Medical School, University of Turin, Turin, Italy.

Background: Gastroenteritis is a common disease in children, characterized by diarrhea, vomiting, abdominal pain, and fever. Co-detection of HBoV with other gastroenteric viruses was reported a lot in patients with acute gastroenteritis.

Methods: This paper presents the real-time RT-PCR Taqman assay for the detection and quantification of HBoV for clinical fecal samples collected from hospitalized children with acute gastroenteritis in Piedmont.

Results: All fecal specimens were tested for the presence of HBoV with specific primers and probe. A total of 17 out of 123 (13.92%) episodes of acute gastroenteritis were associated with HBoV genomic detection with median viral load 6864.75±19784.79 genomes/mg fecal specimens. Among the 17 HBoV-positive cases, 11 were also positive for other viral pathogens, including rotavirus (n = 2), astrovirus (n=1), norovirus GII (n=6), norovirus GI (n=2) . Two cases were positive for more than one virus including norovirus GII and norovirus GI (n=1) and rotavirus, sapovirus and astrovirus (n=1). A higher detection of HBoV infections was observed in winter, and peaking in February.

Conclusions: Although HBoV is suspected to be responsible for gastroenteritis in children, our data showed that this association was uncertain since none difference was observed in term of viral load in the group with single infection of HBoV and group of coinfections with other viral agent.
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http://dx.doi.org/10.23736/S0026-4946.18.05365-3DOI Listing
January 2019

Transcriptional Activity of Human Endogenous Retroviruses in Response to Prenatal Exposure of Maternal Cigarette Smoking.

Am J Perinatol 2019 08 30;36(10):1060-1065. Epub 2018 Nov 30.

Department of Public Health and Pediatric Sciences, Citoimmunodiagnostics Laboratory, University of Turin, Medical School, Turin, Italy.

Objective: Transcription of human endogenous retrovirus (HERV) elements is usually suppressed by epigenetic factors such as DNA methylation and heterochromatin silencing by histone modifications. There is an association between maternal smoking during pregnancy and DNA methylation levels in placental tissue and in DNA from cord blood.

Study Design: We assessed the transcriptional activity of HERV-H, HERV-K, and HERV-W in umbilical cord blood from 47 term babies unexposed to tobacco smoke in utero and 23 term babies exposed to tobacco smoke in utero.

Results: In our population, the HERV-H, HERV-K, and HERV-W families were always transcriptionally active, and the levels of all HERVs (H, K, W) were significantly higher in unexposed than smoke-exposed babies.

Conclusion: This study provides preliminary information about the transcriptional activity of HERV-H, HERV-K, and HERV-W families in human umbilical cord blood.
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http://dx.doi.org/10.1055/s-0038-1675768DOI Listing
August 2019

Lack of detection of Cutavirus DNA using PCR real time in cutaneous T-cell lymphomas.

G Ital Dermatol Venereol 2020 Dec 29;155(6):772-774. Epub 2018 Oct 29.

Unit of Infectious Diseases, Department of Pediatrics, Regina Margherita Children's Hospital, University of Turin, Turin, Italy.

Background: A novel human protoparvovirus named Cutavirus has been discovered. We investigated the presence of Cutavirus in a sample of Cutaneous T-cell lymphomas by using PCR real time TaqMan® (Thermo Fisher Scientific, Waltham, MA, USA).

Methods: In total, 55 CTCL samples were analyzed using a TaqMan® Real time PCR on a 7500 ABI instrument. All of these shown internal control amplification.

Results: The presence of Cutavirus DNA corresponding was examined. CuV DNA sequences were not detected in any skin specimen.

Conclusions: The role of Cutaviruses in cutaneous cancers remains to be investigated.
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http://dx.doi.org/10.23736/S0392-0488.18.06161-8DOI Listing
December 2020

HERV-E expression in peripheral mononuclear cells of patients with psoriasis.

Ital J Dermatol Venerol 2021 Feb 24;156(1):42-45. Epub 2018 Sep 24.

Section of Dermatology, Department of Medical Sciences, University of Turin, Turin, Italy.

Background: Psoriasis is a common inflammatory skin disease characterized by uncontrolled proliferation of keratinocytes and recruitment of T lymphocytes into the skin. Possible triggers for psoriasis have been attributed to drugs or pathogens such as bacteria and possibly virus. Human endogenous retroviruses (HERVs) might play a role in triggering these antiviral immune responses, since the role of HERVs in the pathogenesis of autoimmune diseases has generated considerable interest. Some studies have also reported an association of HERV-E and psoriasis. None of them investigate the HERV-E expression in peripheral blood of psoriasis. All these considerations have prompted us to perform a survey for HERV-E expression in PBMC from psoriatic patients.

Methods: Peripheral blood mononuclear cells from 69 psoriatic patients were analyzed. Total RNA was extracted and amplified with reverse transcription polymerase chain reaction. Results were compared with those obtained in a cohort of 20 healthy donors.

Results: HERV E was expressed in all samples analyzed but the level of expression was much lower in the psoriasis that in HC P<0.0001.

Conclusions: The reasons for the unexpected, low levels of HERV expression in psoriatic patients are unclear and might be in part a consequence of antiviral defense mechanisms.
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http://dx.doi.org/10.23736/S0392-0488.18.06133-3DOI Listing
February 2021

Prevalence of human cosavirus and saffold virus in young children with gastroenteritis, Northern Italy.

Minerva Pediatr 2018 Jul 18. Epub 2018 Jul 18.

Department of Public Health and Pediatric Sciences, Citoimmunodiagnostics Laboratory, University of Turin, Medical School, Turin, Italy -

Background: Gastroenteritis is a common disease in children, characterized by diarrhea, vomiting, abdominal pain, and fever. Human Cosavirus (HCoSV) and Saffold virus (SAFV) both have a worldwide distribution. Both viruses have been detected in the stools of patients with acute gastroenteritis in several countries.

Methods: In order to provide more insights into the epidemiology of enteric viruses that are not included usually in routine diagnostic tests, cases of childhood sporadic gastroenteritis of unknown etiology requiring hospital admission in Turin, Italy, during December 2014 to November 2015, were screened for HCoSV and SAFV.

Results: A total of 1 out of 164 (0.6%) episodes of acute gastroenteritis were associated with SAFV genomic detection. Among the 1 SAFV-positive cases, 1 were also positive for adenovirus. The patient positive for SAFV don't present diarrheal episodes but vomiting. HCoSV was not detected in any of the samples.

Conclusions: In conclusion, this study presents the current epidemiological data regarding the two viruses, HCoSV and SAFV, circulating in pediatric patients admitted to hospital with acute gastroenteritis in Turin, Italy.
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http://dx.doi.org/10.23736/S0026-4946.18.05219-2DOI Listing
July 2018

Prevalence and Clinical Profile of Human Salivirus in Children with Acute Gastroenteritis in Northern Italy, 2014-2015.

Intervirology 2018 18;61(1):49-52. Epub 2018 Jul 18.

Citoimmunodiagnostics Laboratory, Department of Public Health and Pediatric Sciences, University of Turin Medical School, Turin, Italy.

Objective: Human Salivirus (SalV) has been associated with gastroenteritis on all continents.

Methods: This paper presents the real-time RT-PCR assay for the detection of SalV in clinical fecal samples collected from 192 hospitalized children with acute gastroenteritis in Piedmont, Italy.

Results: The most commonly detected virus was Norovirus genogroup II (GII) (33.8%), followed by Rotavirus (21.3%), Sapovirus (10.9%), Parechovirus (8%), Norovirus GI (6.7%), and Adenovirus (1%). PCR detected SalV in 1 (0.5%) subject.

Conclusions: Our data show that the detection rate of SalV in diarrheal children (0.5%) is lower than that observed in other countries, where it is reported in diarrheal children in 8.6-1.2% of patients.
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http://dx.doi.org/10.1159/000490568DOI Listing
December 2018

Is HERV-K and HERV-W Expression Regulated by miR-155 in Kidney Transplant Patients with Human Cytomegalovirus Infection?

Intervirology 2018 10;61(1):23-29. Epub 2018 Jul 10.

Laboratory of Molecular Virology, Maggiore della Carità Hospital, Novara, Italy.

According to the latest update, 2,578 unique mature micro-RNAs (miRNAs) are currently annotated in the human genome and participate in the regulation of multiple events, such as cellular proliferation or apoptosis. A previous study analyzing global miRNA expression patterns in GH cells (high human endogenous retrovirus, HERV, K vs. low) showed that 2 miRNAs (miR-663 and miR-638) are differentially regulated and exhibit expression parallel to that of HERV-K. The aim of this study was to evaluate HERV-K and -W pol gene and miR-155 expression in kidney transplant recipients and the possible relationship between them. The comparison between kidney transplant patients negative for human cytomegalovirus (HCMV) infection and positive patients showed a significant difference in terms of miR-155 expression (p = 0.0111). We demonstrated that HERV-K and -W pol gene expression was significantly higher in CMV-infected kidney transplant recipients versus those not infected as previously reported by our groups. Our correlation data suggest that miR-155 are not directly involved in regulating the HERV notwithstanding that we together observed increased expression of HERV-K and -W and diminished expression of miR-155 in HCMV-infected human kidney transplant recipients.
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http://dx.doi.org/10.1159/000490057DOI Listing
December 2018

DNA from Human Polyomaviruses, MWPyV, HPyV6, HPyV7, HPyV9 and HPyV12 in Cutaneous T-cell Lymphomas.

Anticancer Res 2018 Jul;38(7):4111-4114

Department of Medical Sciences, Dermatology Section, University of Turin, Turin, Italy.

Background/aim: The etiopathogenesis of mycosis fungoides and Sézary syndrome remains obscure. Different viruses have been proposed to have a role in the etiopathogenesis of cutaneous T-cell lymphomas (CTCL). In the present study, the presence of five recently discovered human polyomaviruses 6 (HPyV6), human polyomaviruses 7 (HPyV7), human polyomaviruses 9 (HPyV9), human polyomaviruses 12 (HPyV12), and Malawi polyomavirus (MWPyV), have been analyzed in 55 CTCL in order to confirm the skin tropism and the possible pathological association of these new polyomaviruses.

Materials And Methods: Human polyomaviruses DNA were amplified from skin lesions were recovered from a total of 55 patients (32 males and 23 females, average age 63±15 years) affected by CTCL.

Results: When assayed for the presence of 5 different HPyVs, (HPyV6, HPyV7, HPyV9, MWPyV, and HPyV12) HPyV9, HPyV10 and HPyV12 DNA sequences were not found in any skin specimens. HPyV6 and 7 DNA was detected in 1/55 (1.8%) of skin specimens.

Conclusion: The low-level presence of HPyV6 and HPyV7 DNA, and lack of detection of polyomaviruses HPyV9, MWPyV and HPyV12 in our series do not support a significant role of these HPyVs subtypes in the etiopathogenesis of skin cancers.
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http://dx.doi.org/10.21873/anticanres.12701DOI Listing
July 2018

The interaction between KDR and interleukin-3 receptor (IL-3R) beta common modulates tumor neovascularization.

Oncogene 2005 Sep;24(42):6394-405

Department of Internal Medicine University of Torino, Corso Dogliotti 14, Torino 10126, Italy.

As vascular endothelial growth factor (VEGF), interleukin-3 (IL-3), released into the tumor microenvironment stimulates motogenic and mitogenic activity of normal and transformed cells. In the present study, we investigate the effects of IL-3 and VEGF on neoplastic vascular growth. Engagement of IL-3 receptor beta common (IL-3R beta c) contributes to both IL-3- and VEGF-induced Rac1 activation, cell migration and in vitro tube-like structure formation as shown by the expression of the dominant-negative IL-3R beta c construct (Delta455). In normal and transformed endothelial cells (EC) as well as in HEK 293 cells expressing KDR and IL-3R, VEGF and IL-3 treatment induces the formation of a KDR/IL-3R beta c complex. Moreover, as shown by the IL-3R Delta455 mutant or by the kinase dead KDR, functional receptors are required for this interaction. Consistent with the contribution of IL-3R beta c in both IL-3- and VEGF-mediated angiogenic signal, a reduced number of vessels inside tumors are found in mice injected with cells expressing the IL-3R Delta455 mutant. Thus, these findings provide a novel mechanism through which IL-3 and VEGF support cell survival and tumor neovascularization.
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http://dx.doi.org/10.1038/sj.onc.1208786DOI Listing
September 2005

{beta}1 Integrin and IL-3R coordinately regulate STAT5 activation and anchorage-dependent proliferation.

J Cell Biol 2005 Mar;168(7):1099-108

Department of Genetics, Biology and Biochemistry, University of Torino, 10126, Torino, Italy.

We previously demonstrated that integrin-dependent adhesion activates STAT5A, a well known target of IL-3-mediated signaling. Here, we show that in endothelial cells the active beta1 integrin constitutively associates with the unphosphorylated IL-3 receptor (IL-3R) beta common subunit. This association is not sufficient for activating downstream signals. Indeed, only upon fibronectin adhesion is Janus Kinase 2 (JAK2) recruited to the beta1 integrin-IL-3R complex and triggers IL-3R beta common phosphorylation, leading to the formation of docking sites for activated STAT5A. These events are IL-3 independent but require the integrity of the IL-3R beta common. IL-3 treatment increases JAK2 activation and STAT5A and STAT5B tyrosine and serine phosphorylation and leads to cell cycle progression in adherent cells. Expression of an inactive STAT5A inhibits cell cycle progression upon IL-3 treatment, identifying integrin-dependent STAT5A activation as a priming event for IL-3-mediated S phase entry. Consistently, overexpression of a constitutive active STAT5A leads to anchorage-independent cell cycle progression. Therefore, these data provide strong evidence that integrin-dependent STAT5A activation controls IL-3-mediated proliferation.
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http://dx.doi.org/10.1083/jcb.200405116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2171831PMC
March 2005

RAGE- and TGF-beta receptor-mediated signals converge on STAT5 and p21waf to control cell-cycle progression of mesangial cells: a possible role in the development and progression of diabetic nephropathy.

FASEB J 2004 Aug 4;18(11):1249-51. Epub 2004 Jun 4.

Department of Internal Medicine, University of Torino, Corso Dogliotti 14, Torino, Italy.

The molecular events associated with acute and chronic exposure of mesangial cells (MC) to hyperglycemia were evaluated. We found that, unlike high glucose (HG) and Amadori adducts, advanced glycation end products (AGE) and transforming growth factor-beta (TGF-beta) induced p21waf expression and accumulation of MC in G0/G1. TGF-beta1 blockade inhibited AGE-mediated collagen production but only partially affected AGE-induced p21waf expression and cell-cycle events, indicating that AGE by binding to AGE receptor (RAGE) per se could control MC growth. Moreover, AGE and TGF-beta treatment led to the activation of the signal transduction and activators of transcription (STAT)5 and the formation of a STAT5/p21SIE2 complex. The role of STAT5 in AGE- and TGF-beta-mediated p21waf expression and growth arrest, but not collagen production, was confirmed by the expression of the dominant negative STAT5 (DeltaSTAT5) or the constitutively activated STAT5 (1*6-STAT5) constructs. Finally, in p21waf-/- fibroblasts both AGE and TGF-beta failed to inhibit cell-cycle progression. A potential in vivo role of these mechanisms was sustained by the increasing immunoreactivity for the activated STAT5 and p21(waf) in kidney biopsies from early to advanced stage of diabetic nephropathy. Our data indicate that AGE- and TGF-beta-mediated signals, by converging on STAT5 activation and p21waf expression, may regulate MC growth.
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http://dx.doi.org/10.1096/fj.03-1053fjeDOI Listing
August 2004

IL-3 affects endothelial cell-mediated smooth muscle cell recruitment by increasing TGF beta activity: potential role in tumor vessel stabilization.

Oncogene 2004 Mar;23(9):1681-92

Department of Internal Medicine, University of Torino, Corso Dogliotti 14, 10126 Torino, Italy.

Interleukin-3 (IL-3) expression by tumor-infiltrating lymphocytes (TILs) and its effects on vessel assembly were evaluated. TILs from 'in situ' human breast cancers expressed CD4/CD25 antigens and IL-3. An injection of Matrigel containing SMC and IL-3 or basic-fibroblast growth factor (bFGF) into SCID mice confirmed the neoangiogenetic effect of both factors. However, in response to IL-3, but not to bFGF, only few SMC became incorporated into the nascent vessels. To evaluate the possibility that signals emanated by the nascent vasculature in the presence of IL-3 may negatively regulate SMC recruitment, conditioned media (CM) from IL-3-treated endothelial cells (EC) or SMC were tested for their biological effects on SMC and EC. CM from IL-3-treated SMC stimulated the migration of EC. In contrast, the migration of SMC was not affected by CM from IL-3-stimulated EC; however, it was greatly enhanced by blocking transforming growth factor beta (TGF beta) activity. TGF beta immunoenzymatic assay demonstrated the following: (i) the absence of TGF beta activity in CM from IL-3-stimulated EC; (ii) a barely detectable TGF beta activity in CM from IL-3-stimulated SMC; and (iii) the presence of TGF beta activity in the supernatants of SMC stimulated with CM from IL-3-, but not from bFGF-stimulated EC. Increased TGF beta mRNA expression was only detected in SMC stimulated with CM from IL-3-treated EC. Finally, the inhibitory signals induced by IL-3 in vivo were abrogated by the addition of the neutralizing TGF beta antibody. Thus, the positive immunostaining for IL-3 by TILs in 'in situ' breast cancers sustains the possibility that early in tumor development, IL-3 can contribute to the chronic immaturity of these vessels.
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http://dx.doi.org/10.1038/sj.onc.1207290DOI Listing
March 2004

Angiopoietin 2 induces cell cycle arrest in endothelial cells: a possible mechanism involved in advanced plaque neovascularization.

Arterioscler Thromb Vasc Biol 2004 Mar 15;24(3):511-8. Epub 2004 Jan 15.

Department of Internal Medicine, University of Torino, Torino, Italy.

Objective: To characterize the molecules and the mechanisms regulating the neoangiogenetic process in advanced atherosclerotic plaques.

Methods And Results: Western blot and immunofluorescence analysis of atherosclerotic specimens demonstrated that unlike neovessels from early lesions that expressed vascular endothelial growth factor (VEGF) and angiopoietin1 (Angio1), vessels from advanced lesions expressed VEGF and angiopoietin 2 (Angio2). Moreover, only few neovessels from advanced lesions showed a positive immunostaining for proliferating cell nuclear antigen. Angio1-elicited and Angio2-elicited intracellular events in endothelial cells (EC) demonstrated that while Angio1 triggered Erk1/Erk2 mitogen activated protein kinases (MAPK) and Akt activation, Angio2 (50 ng/mL) induced STAT5 activation and p21waf expression and increased the fraction of cells in G1. Both Angio2-mediated events were abrogated by expressing a dominant negative STAT5 construct (DeltaSTAT5). Consistent with the expression of Angio2 in neovessels of advanced lesions a transcriptionally active STAT5 was detected. Moreover, co-immunoprecipitation experiments revealed the presence of a STAT5/Tie2 molecular complex in neointima vessels from advanced, but not from early, lesions.

Conclusions: In advanced lesions, the activation of the Tie2-mediated STAT5 signaling pathway may negatively regulate vessel growth.
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http://dx.doi.org/10.1161/01.ATV.0000116864.86607.35DOI Listing
March 2004
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