<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

Donne e società in Sardegna: eredità e mutamento: materiali e strumenti di ricerca

Negli anni 1979/80, per una esigenza di confronto interdisciplinare sulla problematica femminile e di apporto alla applicazione corretta delle leggi che riguardavano in particolare le donne, viene organizzato dalle docenti della Facoltà di Magistero per le discipline di Storia Contemporanea, Istituzioni di Diritto Pubblico e Legislazione Scolastica, Psicologia, Pedagogia e Antropologia Culturale, il lO Seminario interdisciplinare con relazioni tenute da gruppi di studio di docenti e studentesse sulle leggi degli anni '70: divorzio, nuovo diritto di famiglia, legge sui consultori, legge di parità sul lavoro, sulla maternità consapevole e i progetti di legge contro la violenza sessuale. In questa direzione, nell'anno seguente, viene individuato il tema dei consultori familiari in Sardegna al fine di mettere a confronto le diverse realtà dell'Isola e, di questo secondo Seminario, vengono poi pubblicati gli atti sul libro Il Consultorio Familiare in Sardegna a cura di Rina Fancellu, edizioni di Iniziative Culturali. Negli anni 1985/86 il gruppo si allarga ed organizza, nell'ambito della collaborazione tra il Dipartimento di Economia, Istituzioni e Società ed il Dipartimento di Storia, un altro Seminario dal titolo Mutamenti istituzionali e culturali nella divisione dei ruoli sessuali dal secondo dopoguerra ad oggi, che è stato riproposto l'anno successivo (1986/87) privilegiando la Sardegna come campo della ricerca. Su questi temi riguardanti i mutamenti istituzionali e culturali nella divisione dei ruoli sessuali in Sardegna, presentiamo questa pubblicazione che comprende sull 'argomento materiali e strumenti di ricerca. Il criterio seguito è quello interdisciplinare tra materiale storico-legislativo, antropologico, psicologico, socioeconomico

Genetic Variation among Pharmacogenes in the Sardinian Population

Pharmacogenetics (PGx) aims to identify the genetic factors that determine inter-individual differences in response to drug treatment maximizing efficacy while decreasing the risk of adverse events. Estimating the prevalence of PGx variants involved in drug response, is a critical preparatory step for large-scale implementation of a personalized medicine program in a target population. Here, we profiled pharmacogenetic variation in fourteen clinically relevant genes in a representative sample set of 1577 unrelated sequenced Sardinians, an ancient island population that accounts for genetic variation in Europe as a whole, and, at the same time is enriched in genetic variants that are very rare elsewhere. To this end, we used PGxPOP, a PGx allele caller based on the guidelines created by the Clinical Pharmacogenetics Implementation Consortium (CPIC), to identify the main phenotypes associated with the PGx alleles most represented in Sardinians. We estimated that 99.43% of Sardinian individuals might potentially respond atypically to at least one drug, that on average each individual is expected to have an abnormal response to about 17 drugs, and that for 27 drugs the fraction of the population at risk of atypical responses to therapy is more than 40%. Finally, we identified 174 pharmacogenetic variants for which the minor allele frequency was at least 10% higher among Sardinians as compared to other European populations, a fact that may contribute to substantial interpopulation variability in drug response phenotypes. This study provides baseline information for further large-scale pharmacogenomic investigations in the Sardinian population and underlines the importance of PGx characterization of diverse European populations, such as Sardinians

ß-endorphin and cortisol levels in plasma and CSF following acute experimental spinal traumas

ß-endorphin and cortisol were measured in cerebrospinal fluid (CSF) and plasma by radioimmunological method (RIA) in two groups of rabbits with spinal cord traumatic injuries at cervical and lumbar levels, respectively with and without concomitant spinal shock and arterial hypotension, and in a group of sham operated animals as controls. The two groups with spinal lesions displayed a significant ß-endorphin increase in CSF, whereas the cortisol level remained unchanged both in the spinal traumatized rabbits and in controls. Both the opioid and the cortisol concentration rose significantly in plasma in all three groups and in particular resulted significantly higher in the cervical traumatized group where spinal trauma was associated with spinal shock and hypotension. However, no significant difference was found when beta-endorphin concentrations in plasma were compared between the sham operated animals and the spinal lumbar traumatized animals without concomitant spinal shock. The results seem to suggest that the ß-endorphin increase in CSF is related to the nervous tissue lesion, while its increase in plasma, like that of cortisol, is due to surgery or other stress factors inherent in the experiment. This independent behaviour of ß-endorphin in plasma and in CSF suggests its different origin in these two compartments

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

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

Overall survival in mCPRC patients treated with Radium-223 in association with bone health agents: a national multicenter study

Radium-223 has demonstrated efficacy in improving overall survival (OS) and in delaying symptomatic skeletal-related events (SREs). Bone Health Agents (BHA), i.e. RANK ligand inhibitor (Denosumab) and bisphosphonate such as zoledronic acid, are indicated to prevent SREs without a clear survival benefit. SREs on patient health have a high impact and it is, therefore, important to consider the role of new therapies with BHA to better understand the involvement of combination therapy. The primary aim of this multicentric study is to assess OS in mCRPC patients treated with Radium-223 in combination with BHA

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

Multiple sequence alignment of the TNASE_3 domain of <i>M. agalactiae</i> PG2^T MAG_5040 with homologous proteins identified in other mycoplasma species, and with the SNc of <i>Staphylococcus aures.</i>

<p>Positions are numbered according to the first amino acid of this domain in each bacterial species. Conserved amino acid residues involved in cations binding and encompassing the catalytic site are shown in boldface. An additionally conserved glycine residue is underlined and in boldface. Asterisks indicate positions conserved in all the bacteria aligned.</p

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