Molecular markers of influenza B lineages and clades

Co-circulation of two influenza B virus lineages, B/Yamagata and B/Victoria, has been recognized since the late 1980s. The assessment of the prevalent lineage and the group of viruses in circulation is of importance in order to decide on the vaccine composition and evaluate its efficacy. The molecular characterization of influenza B viruses in circulation has been the aim of this study; this was approached by identifying and locating nucleotide substitutions in the influenza B virus hemagglutinin (HA) and neuraminidase (NA), specific for the lineage and/or clade. By the alignment of 3456 sequences from the influenza GISAID EpiFlu database, a high number of lineage- and group-specific nucleotide positions have been observed in the HA gene, but not in the NA gene. Additionally, an RT-PCR method has been developed, applicable directly to clinical specimens, which amplifies a short HA region that includes a group of unique molecular signatures. Twenty eight influenza B virus-positive respiratory specimens, collected in Tuscany in the seasons 2012–2013 and 2013–2014, were analyzed. The results revealed two clearly distinguishable patterns: one, more frequent, was characterized by all of the nucleotide changes associated with the B/Yamagata lineage (in most cases of Group 2), whereas the other exhibited all of the changes associated with the B/Victoria lineage. It can be concluded that the analysis of this short HA sequence can permit a rapid, highly sensitive determination of influenza B virus lineages and clades

Human parvovirus PARV4 DNA in tissues from adult individuals: a comparison with human parvovirus B19 (B19V)

<p>Abstract</p> <p>Background</p> <p>PARV4 is a new member of the Parvoviridae family not closely related to any of the known human parvoviruses. Viremia seems to be a hallmark of PARV4 infection and viral DNA persistence has been demonstrated in a few tissues. Till now, PARV4 has not been associated with any disease and its prevalence in human population has not been clearly established. This study was aimed to assess the tissue distribution and the ability to persist of PARV4 in comparison to parvovirus B19 (B19V).</p> <p>Results</p> <p>PARV4 and B19V DNA detection was carried out in various tissues of individuals without suspect of acute viral infection, by a real time PCR and a nested PCR, targeting the ORF2 and the ORF1 respectively. Low amount of PARV4 DNA was found frequently (>40%) in heart and liver of adults individuals, less frequently in lungs and kidneys (23,5 and 18% respectively) and was rare in bone marrow, skin and synovium samples (5,5%, 4% and 5%, respectively). By comparison, B19V DNA sequences were present in the same tissues with a higher frequency (significantly higher in myocardium, skin and bone marrow) except than in liver where the frequency was the same of PARV4 DNA and in plasma samples where B19V frequency was significantly lower than that of PARV4</p> <p>Conclusions</p> <p>The particular tropism of PARV4 for liver and heart, here emerged, suggests to focus further studies on these tissues as possible target for viral replication and on the possible role of PARV4 infection in liver and heart diseases. Neither bone marrow nor kidney seem to be a common target of viral replication.</p

Chronic periodontitis and immunity, towards the implementation of a personalized medicine: A translational research on gene single nucleotide polymorphisms (SNPs) linked to chronic oral dysbiosis in 96 caucasian patients

Chronic periodontitis (CP) is a complex pathology with a significant impact worldwide causing bone loss. Oral dysbiosis is a highly inflammatory condition associated to a long-term insulting infection and represents an underestimated CP key factor associated with an imbalance of pro-inflammatory and anti-inflammatory gene responses. The presence of a single nucleotide polymorphisms (SNPs) in the promoter region of interleukin 10 (IL-10) gene-1082,-819, and-592 was a possible determinant cause. This translational research aimed to provide outcomes on the role of IL-10 gene expression in bone loss diseases in patients affected by CP. Caucasian patients (n = 96) affected by CP were recruited from the Italian population. The subgingival samples were collected using the Bacterial Periodontal Assessment by Biomolecular Diagnostic® and the characterization of a set of 15 bacterial DNA responsible of periodontitis was performed by real-time multiplex PCR. In addition, two viruses, Epstein-Barr Virus (EBV) and Herpes Simplex Virus 1 (HSV-1), and a pathogenic fungi (Candida albicans) were included as a part of our panel. Our results confirmed an existing association between IL-10 gene polymorphisms and polymorphism of tumor necrosis factor alpha (TNFff), interleukin 1α-β-RN (IL-1α-β-RN), collagen type-l alpha (COLIA1), and vitamin D receptor (VDRs) genes in CP. Further studies are needed to improve diagnosis and endorse more effective therapeutic procedures for periodontal disease

Studio dell'infezione persistente da Erythovirus B19

Il parvovirus B19 (VB19), appartenente al genere Erythrovirus, della famiglia Parvoviridae, comprende, secondo dati recenti, 3 genotipi: il genotipo 1 ha come prototipo il parvovirus umano B19; i genotipi 2 e 3 differiscono dal primo e tra di loro per circa il 12% della sequenza dell’intero genoma. Gli studi finora condotti riguardano il genotipo 1, mentre scarse sono, per ora, le conoscenze concernenti gli altri due genotipi. In particolare, non è ancora nota la loro diffusione né il loro ruolo patogenetico (se diverso da quello del genotipo 1) e la loro capacità di persistere. Inoltre, la scoperta dei genotipi 2 e 3 comporta problemi di adeguamento delle metodologie diagnostiche, comunemente impiegate per la diagnosi di infezione da VB19. 1. Determinare la diffusione dei 3 genotipi di VB19, il loro tropismo tessutale e la loro capacità di dare infezioni persistenti. 2. Valutare la capacità del VB19 di infettare fibroblasti cutanei di soggetti normali, come possibile sede dell’infezione persistente. Sono stati analizzati 117 campioni di biopsie tessutali provenienti da pazienti con patologie non associate all’infezione da VB19. Tra questi, 38 biopsie cutanee, 38 aspirati midollari, 30 biopsie sinoviali e 11 biopsie cardiache. Inoltre, nello studio sono stati inclusi 97 campioni di siero provenienti dagli stessi pazienti. I campioni sono stati analizzati medianti due PCR di consenso in grado di amplificare tutti e tre i genotipi e mediante 2 PCR genotipo-specifiche e, in alcuni casi, mediante sequenziamento. Per l’infezione sperimentale di fibroblasti umani, sono state utilizzate colture primarie di fibroblasti cutanei (FU) di soggetti sani e culture primarie di fibroblasti ottenuti dalla cute di soggetti affetti da Sclerosi sistemica oltre a un controllo positivo rappresentato dall’infezione della linea cellulare UT7-Epo. Dopo l’esposizione al virus, le cellule sono state incubate per 2h, 24h, 48h e 6 giorni. A questi tempi, sono stati valutati, come indici di infezione, la presenza di mRNA per le proteine strutturali (VP1) e non strutturali (NS1) del VB19 mediante RT-PCR, e del DNA virale mediante ibridazione in situ (ISH). Complessivamente, il DNA di Erythrovirus è stato dimostrato in 8/38 (21%) campioni di midollo, in 29/38 (76%) biopsie cutanee, in 19/30 (63%) biopsie sinoviali e in 7/11 (64%) biopsie cardiache. Complessivamente, il 63% dei campioni positivi era di genotipo 2, il 31 % era di genotipo 1, mentre rara è risultata la presenza del genotipo 3. Per quanto riguarda l’infezione dei fibroblasti in vitro, RNA messaggeri virali per la NS1 e per le proteine capsidiche sono risultati presenti a 24h, 48h e 6 giorni dopo l’esposizione al virus. Nell’ ISH, invece, non sono state visualizzate cellule positive nelle colture FU, a differenza di quanto osservato nelle cellule UT7, usate come controllo In conclusione, il VB19 persiste nei tessuti, dopo l’infezione acuta, con frequenza molto elevata. Il genotipo più frequentemente presente nei tessuti è il 2, mentre rarissimo è risultato il genotipo 3. Per quanto riguarda l’infezione sperimentale, i risultati ottenuti indicano che i fibroblasti possono essere infettati dal VB19. L’infezione di un numero basso di cellule e la modesta attività del virus in tali cellule potrebbe farne un bersaglio per la persistenza del virus

Different behavior of erythrovirus B19 and torquetenovirus in response to a single step of albumin purification

12noneBACKGROUND: The safety of human serum albumin (HSA) is of special interest with respect to virus transmission because of the wide use of this blood product as a therapeutic agent and also, added to other products, as an excipient or a stabilizer. Conflicting data are reported concerning HSA contamination by small, naked viruses such as the erythrovirus B19 (B19V) and the anellovirus torquetenovirus (TTV). This study has been performed to assess the effect of the HSA purification procedures on the viral contamination. STUDY DESIGN AND METHODS: Known concentrations of B19V and TTV virus were spiked in raw Fraction V, the starting material from fractionated human plasma for HSA purification, which was subsequently submitted to the depth filtration procedure. After spiking, B19V and TTV genome copies were determined by real-time quantitative polymerase chain reaction assays in the mixture at the end of Fraction V dissolution, to determine the virus concentration achieved, in the HSA solution after the filtration step, in the filtered postwashing fluid, and in the supernatant of resuspended Celite. RESULTS: B19V was completely adsorbed by the Celite used as a filter aid in the depth filtration process and was thus undetectable in the resulting HSA-containing fraction. In contrast, in 2 out of 3 experiments, TTV was detected in all samples. CONCLUSION: The different behavior of the two viruses might be a reflection of their different surface charge.mixedAzzi, Alberta; Maggi, Fabrizio; Zakrzewska, Krystyna; Menconi, Maria Carla; Di Pietro, Niccolò; Salotti, Vittorio; Farina, Claudio; Andreoli, Elisabetta; Fiorentino, Bruno; Angelini, Cristina; Corcioli, Fabiana; Bendinelli, MauroAzzi, Alberta; Maggi, Fabrizio; Zakrzewska, Krystyna; Menconi, Maria Carla; Di Pietro, Niccolò; Salotti, Vittorio; Farina, Claudio; Andreoli, Elisabetta; Fiorentino, Bruno; Angelini, Cristina; Corcioli, Fabiana; Bendinelli, Maur

Chronic Periodontitis and Immunity, Towards the Implementation of a Personalized Medicine: A Translational Research on Gene Single Nucleotide Polymorphisms (SNPs) Linked to Chronic Oral Dysbiosis in 96 Caucasian Patients

Chronic periodontitis (CP) is a complex pathology with a significant impact worldwide causing bone loss. Oral dysbiosis is a highly inflammatory condition associated to a long-term insulting infection and represents an underestimated CP key factor associated with an imbalance of pro-inflammatory and anti-inflammatory gene responses. The presence of a single nucleotide polymorphisms (SNPs) in the promoter region of interleukin 10 (IL-10) gene −1082, −819, and −592 was a possible determinant cause. This translational research aimed to provide outcomes on the role of IL-10 gene expression in bone loss diseases in patients affected by CP. Caucasian patients (n = 96) affected by CP were recruited from the Italian population. The subgingival samples were collected using the Bacterial Periodontal Assessment by Biomolecular Diagnostic® and the characterization of a set of 15 bacterial DNA responsible of periodontitis was performed by real-time multiplex PCR. In addition, two viruses, Epstein–Barr Virus (EBV) and Herpes Simplex Virus 1 (HSV-1), and a pathogenic fungi (Candida albicans) were included as a part of our panel. Our results confirmed an existing association between IL-10 gene polymorphisms and polymorphism of tumor necrosis factor alpha (TNFα), interleukin 1α-β-RN (IL-1α-β-RN), collagen type-l alpha (COLIA1), and vitamin D receptor (VDRs) genes in CP. Further studies are needed to improve diagnosis and endorse more effective therapeutic procedures for periodontal disease