The Association of Childhood Asthma and Helicobacter pylori Infection in Sardinia

Background: It has been suggested that Helicobacter pylori infection might reduce the risk of atopic conditions, such as asthma, in childhood. This risk reduction could relate to the "hygiene hypothesis" which proposes an association between childhood exposure to microbes and risk of atopy. Objectives: To examine the association between Hp infection and childhood acquired asthma in Sardinia. Patients and Methods: Children from Northern Sardinia who were between the ages of 10 months to 6 years and were screened for Hp infection in 1994-1995 using IgG serology, were asked in 2012, whether they had developed asthma and/or allergic disease in pediatric age, using the global initiative on asthma guidelines questionnaire. Results: A total of 64 children participated in the study. The sero-positivity for Hp infection was 14.1%. Eleven (17.2%) children had a confirmed diagnosis of asthma with onset before the age of 5 years, 85.9% were Hp negative and 14.1% Hp positive. Eight children of the 53 without asthma were Hp positive (15%) compare to one children positive for the infection of the 11 patients (0.09%) with asthma (8/53 vs. 1/11; P = 0.6). The majority of children (73%) were from urban areas and 43.8% had a family history of asthma or allergies. Multiple logistic regression analysis was not able to find a studied variable, including Hp infection, significantly associated with pediatric asthma. Conclusions: Our results speak against Hp infection itself playing a role to protect from the risk to develop childhood asthma although household hygiene was not directly assessed

<i>Mycoplasma agalactiae</i> MAG_5040 is a Mg²⁺-dependent, sugar-nonspecific SNase recognised by the host humoral response during natural infection

In this study the enzymatic activity of Mycoplasma agalactiae MAG_5040, a magnesium-dependent nuclease homologue to the staphylococcal SNase was characterized and its antigenicity during natural infections was established. A UGA corrected version of MAG_5040, lacking the region encoding the signal peptide, was expressed in Escherichia coli as a GST fusion protein. Recombinant GST-MAG_5040 exhibits nuclease activity similar to typical sugar-nonspecific endo- and exonucleases, with DNA as the preferred substrate and optimal activity in the presence of 20 mM MgCl2 at temperatures ranging from 37 to 45uC. According to in silico analyses, the position of the gene encoding MAG_5040 is consistently located upstream an ABC transporter, in most sequenced mycoplasmas belonging to the Mycoplasma hominis group. In M. agalactiae, MAG_5040 is transcribed in a polycistronic RNA together with the ABC transporter components and with MAG_5030, which is predicted to be a sugar solute binding protein by 3D modeling and homology search. In a natural model of sheep and goats infection, anti-MAG_5040 antibodies were detected up to 9 months post infection. Taking into account its enzymatic activity, MAG_5040 could play a key role in Mycoplasma agalactiae survival into the host, contributing to host pathogenicity. The identification of MAG_5040 opens new perspectives for the development of suitable tools for the control of contagious agalactia in small ruminants

Structured reporting for fibrosing lung disease: a model shared by radiologist and pulmonologist

Objectives: To apply the Delphi exercise with iterative involvement of radiologists and pulmonologists with the aim of defining a structured reporting template for high-resolution computed tomography (HRCT) of patients with fibrosing lung disease (FLD). Methods: The writing committee selected the HRCT criteria\ue2\u80\u94the Delphi items\ue2\u80\u94for rating from both radiology panelists (RP) and pulmonology panelists (PP). The Delphi items were first rated by RPs as \ue2\u80\u9cessential\ue2\u80\u9d, \ue2\u80\u9coptional\ue2\u80\u9d, or \ue2\u80\u9cnot relevant\ue2\u80\u9d. The items rated \ue2\u80\u9cessential\ue2\u80\u9d by &lt; 80% of the RP were selected for the PP rating. The format of reporting was rated by both RP and PP. Results: A total of 42 RPs and 12 PPs participated to the survey. In both Delphi round 1 and 2, 10/27 (37.7%) items were rated \ue2\u80\u9cessential\ue2\u80\u9d by more than 80% of RP. The remaining 17/27 (63.3%) items were rated by the PP in round 3, with 2/17 items (11.7%) rated \ue2\u80\u9cessential\ue2\u80\u9d by the PP. PP proposed additional items for conclusion domain, which were rated by RPs in the fourth round. Poor consensus was observed for the format of reporting. Conclusions: This study provides a template for structured report of FLD that features essential items as agreed by expert thoracic radiologists and pulmonologists

Mycoplasma agalactiae MAG_5040 is a Mg2+-Dependent, Sugar-Nonspecific SNase Recognised by the Host Humoral Response during Natural Infection

none12noIn this study the enzymatic activity of Mycoplasma agalactiae MAG_5040, a magnesium-dependent nuclease homologue to the staphylococcal SNase was characterized and its antigenicity during natural infections was established. A UGA corrected version of MAG_5040, lacking the region encoding the signal peptide, was expressed in Escherichia coli as a GST fusion protein. Recombinant GST-MAG_5040 exhibits nuclease activity similar to typical sugar-nonspecific endo- and exonucleases, with DNA as the preferred substrate and optimal activity in the presence of 20 mM MgCl2 at temperatures ranging from 37 to 45°C. According to in silico analyses, the position of the gene encoding MAG_5040 is consistently located upstream an ABC transporter, in most sequenced mycoplasmas belonging to the Mycoplasma hominis group. In M. agalactiae, MAG_5040 is transcribed in a polycistronic RNA together with the ABC transporter components and with MAG_5030, which is predicted to be a sugar solute binding protein by 3D modeling and homology search. In a natural model of sheep and goats infection, anti-MAG_5040 antibodies were detected up to 9 months post infection. Taking into account its enzymatic activity, MAG_5040 could play a key role in Mycoplasma agalactiae survival into the host, contributing to host pathogenicity. The identification of MAG_5040 opens new perspectives for the development of suitable tools for the control of contagious agalactia in small ruminants. © 2013 Cacciotto et al.noneCacciotto C.; Addis M.F.; Coradduzza E.; Carcangiu L.; Nuvoli A.M.; Tore G.; Dore G.M.; Pagnozzi D.; Uzzau S.; Chessa B.; Pittau M.; Alberti A.Cacciotto C.; Addis M.F.; Coradduzza E.; Carcangiu L.; Nuvoli A.M.; Tore G.; Dore G.M.; Pagnozzi D.; Uzzau S.; Chessa B.; Pittau M.; Alberti A

Mycoplasma agalactiae MAG_5040 is a Mg2+-dependent, sugar-nonspecific SNase recognised by the host humoral response during natural infection.

In this study the enzymatic activity of Mycoplasma agalactiae MAG_5040, a magnesium-dependent nuclease homologue to the staphylococcal SNase was characterized and its antigenicity during natural infections was established. A UGA corrected version of MAG_5040, lacking the region encoding the signal peptide, was expressed in Escherichia coli as a GST fusion protein. Recombinant GST-MAG_5040 exhibits nuclease activity similar to typical sugar-nonspecific endo- and exonucleases, with DNA as the preferred substrate and optimal activity in the presence of 20 mM MgCl2 at temperatures ranging from 37 to 45°C. According to in silico analyses, the position of the gene encoding MAG_5040 is consistently located upstream an ABC transporter, in most sequenced mycoplasmas belonging to the Mycoplasma hominis group. In M. agalactiae, MAG_5040 is transcribed in a polycistronic RNA together with the ABC transporter components and with MAG_5030, which is predicted to be a sugar solute binding protein by 3D modeling and homology search. In a natural model of sheep and goats infection, anti-MAG_5040 antibodies were detected up to 9 months post infection. Taking into account its enzymatic activity, MAG_5040 could play a key role in Mycoplasma agalactiae survival into the host, contributing to host pathogenicity. The identification of MAG_5040 opens new perspectives for the development of suitable tools for the control of contagious agalactia in small ruminants

Host cell tropism, genome characterization, and evolutionary features of OaPV4, a novel Deltapapillomavirus identified in sheep fibropapilloma

Investigating papillomavirus (PV) diversity is crucial to fully comprehend pathogenicity, genetic features, and evolution of taxa hosted by domestic and wild animal species. This study reports the identification of OaPV4, a novel ovine PV type within Deltapapillomaviruses 3. The study of OaPV4 genomic features combined to in situ hybridization and immunohistochemistry investigations allowed extrapolating several general biological features of ovine PVs, such as their cellular tropism, pathogenicity, and evolutionary history. Based on results, ovine PVs can be grouped into a polyphyletic ancient group of viruses, which splits in two main subgroups having peculiar cellular tropism and pathogenicity. Results add up to animal PV diversity and are crucial to future studies aimed to investigate the correlation between animal PV and cutaneous benign and malign proliferations

MAG_5040 western blotting reactivity with sera collected from naturally infected hosts, and indirect detection of homologs in selected mycoplasma species.

<p>200 ng of cleaved rMAG_5040 were run in single well in 10% polyacrylamide gels and probed with sera collected alternatively from Piedmont goats naturally infected with <i>M. agalactiae</i> (A) or from sheep selected from an outbreak of contagious agalactia occurred in Sicily (B). In A, odd lanes (1 to 15) identify sera collected from 8 different goats. Even numbers (2 to 16) indicate sera taken from the same goats at two weeks distance. In B lanes 1 to 10 were probed with sera collected from 10 sheep. Neg in all panels designates lanes in which a pool of negative sera was loaded. (C) Reactivity of of cleaved rMAG_5040 (200 ng) with rabbit hyperimmune sera raised against <i>M. agalactiae</i> PG2^T (1), <i>M. mycoides</i> subsp. <i>capri</i> PG3 (2), <i>M. capricolum</i> subsp. <i>capricolum</i> CK (3), <i>M. arginini</i> G230 (4), <i>M. canadense</i> C275 (5), <i>M. ovipneumoniae</i> Y98 (6), <i>M. putrefaciens</i> KS1 (7), <i>M. mycoides</i> subsp. <i>capri</i> LC (8), and <i>M. capricolum</i> subsp. <i>capripneumoniae</i> (9). Lane 10 equals lane 1 except that no primary antibody was used.</p

Nuclease activity and substrate specificity of recombinant GST-MAG_5040.

<p>Approximately 4 µg of rGST-MAG_5040 were incubated with calf thymus dsDNA (top left panel), closed circular plasmid DNA (pGEX-2T/rMAG_5040, top right panel), phage M13 ssDNA (bottom left panel), or <i>E. coli</i> total RNA (bottom right panel). An aliquot of each reaction mixture was analysed at different times, as indicated for each lane. C indicates endpoint reaction of the negative undigested control (GST only). Both 1 Kb (upper panels) and 1 Kb Plus (bottom panels) DNA ladders were used (Invitrogen) and indicated as MW in far left lanes of each panel.</p