PMA E BIOLOGIA FORENSE IN AUSILIO DEL PUBBLICO MINISTERO

PMA E BIOLOGIA FORENSE IN AUSILIO DEL PUBBLICO MINISTERO Carra E., Di Natale M.V. Dipartimento di Scienze Tecnologie Molecolari Biomolecolari (STEMBIO)– Università di Palermo Edifio 16, viale delle Scienze.– [email protected] Nell’ambito di attività svolta su incarico dell’Autorità Giudiziaria si è proceduto all’inventario del contenuto di fusti per la crioconservazione di gameti ed embrioni sottoposti a sequestro e custoditi presso i locali di quattro centri di biologia della riproduzione per “verificarne la rispondenza alle registrazioni cartacee ed informatiche acquisite agli atti”. Si rendeva, altresì, necessaria analisi genetica su materiale biologico, costituito da embrione umano degenerato e la ricostruzione del percorso dei gameti e/o embrioni dei pazienti che avevano ricevuto trattamenti di riproduzione medicalmente assistita per “evidenziare eventuali irregolarità significative ai sensi della normativa vigente in materia di PMA”. Trattasi delle prime indagini commissionate sul territorio Nazionale che vedono impegnato il biologo forense in uno dei settori di frontiera della medicina legata alle nuove tecnologie. Con il pronunciamento della Consulta nel maggio 2009, infatti, l’impianto della L. nr. 40/2004 non è cambiato. Si possono creare embrioni solo a fini di procreazione e restano i divieti alla loro crioconservazione, soppressione e alla selezione a fini eugenetici. Il numero di embrioni da creare resta quello “strettamente necessario” e sarà il medico a stabilire caso per caso, quel numero “strettamente necessario”. Rimangono eccezioni alla crioconservazione, con una maggiore attenzione alla tutela della salute della donna. Mediante annotazione del numero dei canestri, dei jonc, delle paillettes e delle indicazioni riportate è stato eseguito il censimento del contenuto dei fusti criogenici (ovociti, seme, embrioni); tuttavia, la vastità della documentazione in sequestro imponeva di definire correttamente il numero di gameti e degli embrioni criopreservati per le specifiche coppie di pazienti. Venivano realizzati programmi per interventi di bonifica su tutte le tabelle dell’anagrafica clienti acquisite in file di Access per ottenere una Tabella Anagrafica per individuare, tra i vari databases dei Centri BDR, quello ad essi corrispondente ed aggiornato alla data del sequestro. Si è poi proceduto ad un 2° censimento dei fusti criogenici predisponendo, a verifica, modulistica per tutto il materiale criopreservato. Si evidenziavano difformità. L’analisi genetica era condotta su embrione umano criopreservato e crioscongelato compromesso in maniera irreversibile nello sviluppo, acquisito al sequestro all’interno di paillette. Alla microscopia non veniva individuato alcun embrione all’interno del capillare, ma la modalità adottata per il recupero del contenuto di esso ha contribuito al successo delle analisi molecolari. Preliminarmente, però, ci si avvaleva di modello biologico animale per valutare fino a che punto il livello di degenerazione cellulare ne potesse impedire il riconoscimento alla microscopia. L’analisi a campione su schede biologiche, cartelle cliniche, registri IVF, ecc.. unita all’informatizzazione di singoli riscontri per tipologia di scheda (data pick up, donna, uomo, ovociti totali, tecnica, ovociti maturi, seme, embrioni, ovociti scartati, embrioni scartati, ovociti congelati, embrioni congelati, embrioni trasferiti, medico, biologo, analisi, note) ha consentito quella ricostruzione di percorso commissionata evidenziando il modus operandi dei sanitari in violazione degli artt.13 e 14 L. nr. 40/2004 cit.

Risk factors for long-stay in an Italian acute psychiatric ward: a 7-year retrospective analysis

Background: In West during the last decades, the phenomenon of "bed blockers" has been more frequently investigated, probably because of increasing economic constraints in the management of public health. According to most authors, the lack of rehabilitation facilities, organizational problems within the hospital, the long wait for medical consultations and diagnostic procedures would be the main causes of "delayed discharge". Early studies were carried out in long-term care, rehabilitation and post-acute geriatric wards. In Psychiatry, the few studies on this topic highlighted a wide range of causes, including both patient conditions and organizational health system problems. In Italy, the problem of psychiatric delayed discharges has become more pressing after the 180 Law, which established the closure of all psychiatric hospitals and implemented psychiatric wards inside General Hospitals to admit only 15 acute patients for a very short period. Purposes: To highlight the phenomenon of long-stay in an acute psychiatric ward and to relate it to demographic, clinical and organizational variables. Methods: The survey was conducted in the 15-bed public psychiatric ward of Modena (Italy). All admissions were retrospectively collected from the database of the Department from 1 January 2005 to 31 December 2011 (3981 hospitalizations with an average stay of 12.49 days). Demographic data, clinical variables, inpatient care problems, discharge programs were statistically related to the duration of admissions (survival analysis: log-rank test, Kaplan-Meier curves). The 3981 hospitalizations were divided into two groups according to the 90° percentile of duration: < 27 days (n=3575) and ≥ 27 days (n=406) and the variables of the two groups were compared (multiple logistic regression). Secondary analysis was conducted on the subgroup of the longest hospitalizations further divided into two groups according to the 90° (from 27 days to < 36 days) and 95° percentile (≥36 days), in order to find out variables related (survival analysis: log-rank test; multiple logistic regression test). Results: The longest hospitalizations (≥27 days) represent 11% of all admissions during the observation period. When all variables are compared to the duration of hospitalizations, most of them are statistically significantly related to the length of hospitalizations, but, when statistical analysis was focused on the comparison between the two groups of the longest hospitalizations, a smaller number of variables (“gender”, “age”, “rehabilitative programs”, “extra-psychiatric clinical activities”, “pharmacotherapy” and “aggressiveness of patient”) were identified by survival analysis as statistically significant correlates of long-stay (log-rank test), whereas only “female gender” and aggressiveness pf patient” were the variables statistically significantly related to the length of hospitalizations evidenced by multivariate logistic regression analysis. Conclusions: Our results suggest that a wide range of factors may be responsible for the delayed discharges in psychiatry as most previous studies have already shown. However, only few factors were related to the longest duration of hospitalization and, among these, aggressiveness was the only one statistically significant correlate to long-stay in all statistic tests. This data confirms the clinical observation that aggressive behaviour can be sufficient by itself to explain the difficulty of discharging

The small heat shock protein B8 (HSPB8) modulates proliferation and migration of breast cancer cells

open12noBreast cancer (BC) is one of the major causes of cancer death in women and is closely related to hormonal dysregulation. Estrogen receptor (ER)-positive BCs are generally treated with anti hormone therapy using antiestrogens or aromatase inhibitors. However, BC cells may become resistant to endocrine therapy, a process facilitated by autophagy, which may either promote or suppress tumor expansion. The autophagy facilitator HSPB8 has been found overexpressed in some BC. Here we found that HSPB8 is highly expressed and differentially modulated by natural or synthetic selective ER modulators (SERMs), in the triple-positive hormone-sensitive BC (MCF-7) cells, but not in triple-negative MDA-MB-231 BC cells. Specific SERMs induced MCF-7 cells proliferation in a HSPB8 dependent manner whereas, did not modify MDA-MB-231 cell growth. ER expression was unaffected in HSPB8-depleted MCF-7 cells. HSPB8 over-expression did not alter the distribution of MCF-7 cells in the various phases of the cell cycle. Conversely and intriguingly, HSPB8 downregulation resulted in an increased number of cells resting in the G0/G1 phase, thus possibly reducing the ability of the cells to pass through the restriction point. In addition, HSPB8 downregulation reduced the migratory ability of MCF-7 cells. None of these modifications were observed, when another small HSP (HSPB1), also expressed in MCF-7 cells, was downregulated. In conclusion, our data suggest that HSPB8 is involved in the mechanisms that regulate cell cycle and cell migration in MCF-7 cells.openPiccolella, Margherita; Crippa, Valeria; Cristofani, Riccardo; Rusmini, Paola; Galbiati, Mariarita; Elena Cicardi, Maria; Meroni, Marco; Ferri, Nicola; Morelli, Federica F; Carra, Serena; Messi, Elio; Poletti, AngeloPiccolella, Margherita; Crippa, Valeria; Cristofani, Riccardo; Rusmini, Paola; Galbiati, Mariarita; Elena Cicardi, Maria; Meroni, Marco; Ferri, Nicola; Morelli, Federica F; Carra, Serena; Messi, Elio; Poletti, Angel

Effectiveness of combination of Mini-and Microsatellite loci to sub-type Mycobacterium avium subsp. paratuberculosis Italian type C isolates

<p>Abstract</p> <p>Background</p> <p><it>Mycobacterium avium </it>subsp. <it>paratuberculosis </it>(Map) is the etiological agent of paratuberculosis. The aim of our study was to combine Mini-and Microsatellite loci analysis in order to explore the effectiveness of this sub-typing method in a group of Map isolates. For this purpose, 84 Italian Type C Map isolates, each from a different cattle herd, were submitted to MIRU-Variable-Number Tandem-Repeats (VNTRs) typing and Short Sequence repeats (SSRs) sequencing. Moreover, the method was used to analyse the variability inside 10 herds (from three to 50 isolates per herd).</p> <p>Results</p> <p>The molecular sub-typing, carried out using three SSR and 10 MIRU-VNTR loci, differentiated the 84 isolates into 33 clusters, reaching a Simpson's Discriminatory Index (SID) value of 0.952 (0.933 to 0.972, 95% confidence intervals). Among all considered loci, six (SSR2, MIRU2, SSR1, SSR8, VNTR3527 and VNTR1067) showed relevant allelic variability. Thirty-eight% of the isolates were clustered into four genotypes, differing from each other for the SSR2 locus. The other isolates, characterised by differences in two or more loci, were spread among the rest of the clusters. The intra-herd analysis revealed more than one genotype in most herds with a similar distribution of clusters.</p> <p>Conclusions</p> <p>Our results revealed the advantage of using both Mini-and Microsatellite approaches for successfully discriminating among Map Type C isolates from the same geographic area, host species and herd. These data suggest that the combination of loci here proposed could be a useful molecular tool for regional epidemiological studies.</p

The Regulation of the Small Heat Shock Protein B8 in Misfolding Protein Diseases Causing Motoneuronal and Muscle Cell Death

Misfolding protein diseases are a wide class of disorders in which the aberrantly folded protein aggregates accumulate in affected cells. In the brain and in the skeletal muscle, misfolded protein accumulation induces a variety of cell dysfunctions that frequently lead to cell death. In motoneuron diseases (MNDs), misfolded proteins accumulate primarily in motoneurons, glial cells and/or skeletal muscle cells, altering motor function. The deleterious effects of misfolded proteins can be counteracted by the activity of the protein quality control (PQC) system, composed of chaperone proteins and degradative systems. Here, we focus on a PQC system component: heat shock protein family B (small) member 8 (HSPB8), a chaperone induced by harmful stressful events, including proteotoxicity. In motoneuron and muscle cells, misfolded proteins activate HSPB8 transcription and enhance HSPB8 levels, which contributes to prevent aggregate formation and their harmful effects. HSPB8 acts not only as a chaperone, but also facilitates the autophagy process, to enable the efficient clearance of the misfolded proteins. HSPB8 acts as a dimer bound to the HSP70 co-chaperone BAG3, a scaffold protein that is also capable of binding to HSP70 (associated with the E3-ligase CHIP) and dynein. When this complex is formed, it is transported by dynein to the microtubule organization center (MTOC), where aggresomes are formed. Here, misfolded proteins are engulfed into nascent autophagosomes to be degraded via the chaperone-assisted selective autophagy (CASA). When CASA is insufficient or impaired, HSP70 and CHIP associate with an alternative co-chaperone, BAG1, which routes misfolded proteins to the proteasome for degradation. The finely tuned equilibrium between proteasome and CASA activity is thought to be crucial for maintaining the functional cell homeostasis during proteotoxic stresses, which in turn is essential for cell survival. This fine equilibrium seems to be altered in MNDs, like Amyotrophic lateral sclerosis (ALS) and spinal and bulbar muscular atrophy (SBMA), contributing to the onset and the progression of disease. Here, we will review how misfolded proteins may affect the PQC system and how the proper activity of this system can be restored by boosting or regulating HSPB8 activity, with the aim to ameliorate disease progression in these two fatal MNDs

Foodborne Salmonellosis in Italy: Characterization of Salmonella enterica Serovar Typhimurium and Monophasic Variant 4,[5],12:i- Isolated from Salami and Human Patients.

Salmonella enterica serovar Typhimurium (STm) and its monophasic variant 4,[5],12:i:- (VMSTm) have been responsible for an increased number of foodborne infections in humans in Europe in recent years. The aim of this study was to investigate the origin of three foodborne salmonellosis outbreaks that occurred in Pavia Province (Lombardy region, northern Italy) in 2010. Phenotypic and genetic characteristics of the STm and VMSTm isolates from patients and from food that were recovered in the framework of the three outbreaks were evaluated through serotyping, phage typing, antimicrobial susceptibility testing, pulsed-field gel electrophoresis (PFGE), and multiple-locus variable-number tandem repeat analysis (MLVA). Salami from three artisan producers, which had all purchased meat from the same slaughterhouse, was the food source of infection in outbreak I. STm isolates were recovered from salami and patients with symptoms of gastroenteritis. These isolates had the same PFGE type and the same rare MLVA profile (3-18-9-NA-211). The same molecular profiles were found in an STm isolate from a salami, which likely was the source of another family outbreak (II). A VMSTm strain with common phenotypic and molecular profiles was isolated from three hospitalized patients and identified as the cause of another putative outbreak (III). During the following 3 years (2011 through 2013), 360 salami produced in Pavia Province were monitored for the presence of S. enterica . In 2011, no STm and VMSTm isolates were recovered from 159 salami tested. During 2012 and 2013, 13.9% of 201 tested salami harbored S. enterica , and half of the isolates were VMSTm, mainly in salami from those artisan producers involved in the previous outbreaks. These isolates were genetically variable, especially in terms of MLVA profiles. The data collected suggest that from 2012, VMSTm has replaced STm in the environments of the salami producers monitored in this study, and these data confirm the dominance of this emergent serovar along the pork supply chain

The economic costs of biological invasions in Central and South America: a first regional assessment

Invasive alien species are responsible for a high economic impact on many sectors worldwide. Nevertheless, there is a scarcity of studies assessing these impacts in Central and South America. Investigating costs of invasions is important to motivate and guide policy responses by increasing stakeholders’ awareness and identifying action priorities. Here, we used the InvaCost database to investigate (i) the geographical pattern of biological invasion costs across the region; (ii) the monetary expenditure across taxa and impacted sectors; and (iii) the taxa responsible for more than 50% of the costs (hyper-costly taxa) per impacted sector and type of costs. The total of reliable and observed costs reported for biological invasions in Central and South America was USD 102.5 billion between 1975 and 2020, but about 90% of the total costs were reported for only three countries (Brazil, Argentina and Colombia). Costs per species were associated with geographical regions (i.e., South America, Central America and Islands) and with the area of the countries in km2. Most of the expenses were associated with damage costs (97.8%), whereas multiple sectors (77.4%), agriculture (15%) and public and social welfare (4.2%) were the most impacted sectors. Aedes spp. was the hyper-costly taxon for the terrestrial environment (costs of USD 25 billion) and water hyacinth (Eichhornia crassipes) was the hyper-costly taxon for the aquatic environment (USD 179.9 million). Six taxa were classified as hyper-costly for at least one impacted sector and two taxa for at least one type of cost. In conclusion, invasive alien species caused billions of dollars of economic burden in Central and South America, mainly in large countries of South America. Costs caused by invasive alien species were unevenly distributed across countries, impacted sectors, types of costs and taxa (hyper-costly taxa). These results suggest that impacted sectors should drive efforts to manage the species that are draining financial sources.info:eu-repo/semantics/publishedVersio

The chaperone HSPB8 reduces the accumulation of truncated TDP-43 species in cells and protects against TDP-43-mediated toxicity

Aggregation of TAR-DNA-binding protein 43 (TDP-43) and of its fragments TDP-25 and TDP-35 occurs in amyotrophic lateral sclerosis (ALS). TDP-25 and TDP-35 act as seeds for TDP-43 aggregation, altering its function and exerting toxicity. Thus, inhibition of TDP-25 and TDP-35 aggregation and promotion of their degradation may protect against cellular damage. Upregulation of HSPB8 is one possible approach for this purpose, since this chaperone promotes the clearance of an ALS associated fragments of TDP-43 and is upregulated in the surviving motor neurones of transgenic ALS mice and human patients. We report that overexpression of HSPB8 in immortalized motor neurones decreased the accumulation of TDP-25 and TDP-35 and that protection against mislocalized/truncated TDP-43 was observed for HSPB8 in Drosophila melanogaster. Overexpression of HSP67Bc, the functional ortholog of human HSPB8, suppressed the eye degeneration caused by the cytoplasmic accumulation of a TDP-43 variant with a mutation in the nuclear localization signal (TDP-43-NLS). TDP-43-NLS accumulation in retinal cells was counteracted by HSP67Bc overexpression. According with this finding, downregulation of HSP67Bc increased eye degeneration, an effect that is consistent with the accumulation of high molecular weight TDP-43 species and ubiquitinated proteins. Moreover, we report a novel Drosophila model expressing TDP-35, and show that while TDP-43 and TDP-25 expression in the fly eyes causes a mild degeneration, TDP-35 expression leads to severe neurodegeneration as revealed by pupae lethality; the latter effect could be rescued by HSP67Bc overexpression. Collectively, our data demonstrate that HSPB8 upregulation mitigates TDP-43 fragment mediated toxicity, in mammalian neuronal cells and flies

Integración de técnicas metrológicas y termográficas para el seguimiento y medición de la influencia de la temperatura en el movimiento del lienzo sur de la nave del crucero de la iglesia del antiguo Convento de San Luis en San Vicente de la Barquera (Cantabria)

El trabajo que se presenta tenía como objetivo la búsqueda de una solución ingenieril que garantizara la consolidación tanto de la cimentación como de los contrafuertes y la fábrica de mampostería del lienzo de la fachada sur del Convento de San Luis, situado en el norte de la península ibérica, más concretamente en San Vicente de la Barquera (España), utilizando para ello las técnicas más apropiadas que puedan asegurar el buen comportamiento futuro de la misma. En este artículo se centra en la integración de varias técnicas geomáticas con el objetivo de controlar geométricamente la buena ejecución de la solución propuesta y, lo que es más importante, la integración en dicho análisis de una variable que no puede pasar desapercibida como es la radiación solar incidente sobre el lienzo sur de la nave del crucero de la Iglesia del antiguo convento de San Luis

The small heat shock protein B8 (HSPB8) efficiently removes aggregating species of dipeptides produced in C9ORF72-related neurodegenerative diseases

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are two neurodegenerative diseases in which similar pathogenic mechanisms are involved. Both diseases associate to the high propensity of specific misfolded proteins, like TDP-43 or FUS, to mislocalize and aggregate. This is partly due to their intrinsic biophysical properties and partly as a consequence of failure of the neuronal protein quality control (PQC) system. Several familial ALS/FTD cases are linked to an expansion of a repeated G4C2 hexanucleotide sequence present in the C9ORF72 gene. The G4C2, which localizes in an untranslated region of the C9ORF72 transcript, drives an unconventional repeat-associated ATG-independent translation. This leads to the synthesis of five different dipeptide repeat proteins (DPRs), which are not âclassicalâ misfolded proteins, but generate aberrant aggregation-prone unfolded conformations poorly removed by the PQC system. The DPRs accumulate into p62/SQSTM1 and ubiquitin positive inclusions. Here, we analyzed the biochemical behavior of the five DPRs in immortalized motoneurons. Our data suggest that while the DPRs are mainly processed via autophagy, this system is unable to fully clear their aggregated forms, and thus they tend to accumulate in basal conditions. Overexpression of the small heat shock protein B8 (HSPB8), which facilitates the autophagy-mediated disposal of a large variety of classical misfolded aggregation-prone proteins, significantly decreased the accumulation of most DPR insoluble species. Thus, the induction of HSPB8 might represent a valid approach to decrease DPR-mediated toxicity and maintain motoneuron viability