Expert-based development of a generic HACCP-based risk management system to prevent critical negative energy balance in dairy herds

The objective of this study was to develop a generic risk management system based on the Hazard Analysis and Critical Control Point (HACCP) principles for the prevention of critical negative energy balance (NEB) in dairy herds using an expert panel approach. In addition, we discuss the advantages and limitations of the system in terms of implementation in the individual dairy herd. For the expert panel, we invited 30 researchers and advisors with expertise in the field of dairy cow feeding and/or health management from eight European regions. They were invited to a Delphi-based set-up that included three inter-correlated questionnaires in which they were asked to suggest risk factors for critical NEB and to score these based on 'effect' and 'probability'. Finally, the experts were asked to suggest critical control points (CCPs) specified by alarm values, monitoring frequency and corrective actions related to the most relevant risk factors in an operational farm setting. A total of 12 experts (40 %) completed all three questionnaires. Of these 12 experts, seven were researchers and five were advisors and in total they represented seven out of the eight European regions addressed in the questionnaire study. When asking for suggestions on risk factors and CCPs, these were formulated as 'open questions', and the experts' suggestions were numerous and overlapping. The suggestions were merged via a process of linguistic editing in order to eliminate doublets. The editing process revealed that the experts provided a total of 34 CCPs for the 11 risk factors they scored as most important. The consensus among experts was relatively high when scoring the most important risk factors, while there were more diverse suggestions of CCPs with specification of alarm values and corrective actions. We therefore concluded that the expert panel approach only partly succeeded in developing a generic HACCP for critical NEB in dairy cows. We recommend that the output of this paper is used to inform key areas for implementation on the individual dairy farm by local farm teams including farmers and their advisors, who together can conduct herd-specific risk factor profiling, organise the ongoing monitoring of herd-specific CCPs, as well as implement corrective actions when CCP alarm values are exceeded

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Impact of diffused versus vasculature targeted DNA damage on the heart of mice depleted of telomeric factor Ft1

: DNA damage is emerging as a driver of heart disease, although the cascade of events, its timing, and the cell types involved are yet to be fully clarified. In this context, the implication of cardiomyocytes has been highlighted, while that of vasculature smooth muscle cells has been implicated but not explored exhaustively. In our previous work we characterized a factor called Ft1 in mice and AKTIP in humans whose depletion generates telomere instability and DNA damage. Herein, we explored the effect of the reduction of Ft1 on the heart with the goal of comparatively defining the impact of DNA damage targeted to vasculature smooth muscle cells to that of diffuse damage. Using two newly generated mouse models, Ft1 constitutively knocked out (Ft1ko) mice, and mice in which we targeted the Ft1 depletion to the smooth muscle cells (Ft1sm22ko), it is shown that both genetic models display cardiac defects but with differences. Both Ft1ko and Ft1sm22ko mice display hypertrophy, fibrosis, and functional heart defects. Interestingly, Ft1sm22ko mice have early milder pathological traits that become manifest with age. Significantly, the defects of Ft1ko mice, including the alteration of the left ventricle and functional heart defects, are rescued by depletion of the DNA damage sensor p53. These results point to Ft1 deficiency as a driver of cardiac disease and show that Ft1 deficiency targeted to vasculature smooth muscle cells generates a pre-pathological profile exacerbated by age

Mutations in GMPPB Presenting with Pseudometabolic Myopathy

Mutations in the guanosine diphosphate mannose (GDP-mannose) pyrophosphorylase B (GMPPB) gene encoding a key enzyme of the glycosylation pathway have been described in families with congenital (CMD) and limb girdle (LGMD) muscular dystrophy with reduced alpha-dystroglycan (\u3b1-DG) at muscle biopsy.Patients typically display a combined phenotype of muscular dystrophy, brain malformations, and generalized epilepsy. However, a wide spectrum of clinical severity has been described ranging from classical CMD presentation to children with mild, yet progressive LGMD with or without intellectual disability. Cardiac involvement, including a long QT interval and left ventricular dilatation, has also been described in four cases.Two missense mutations in GMPPB gene, one novel and one already reported, have been identified in a 21-year-old man presenting with elevated CK (38,650 UI/L; normal values <150 UI/L) without overt muscle weakness. Major complaints included limb myalgia, exercise intolerance, and several episodes of myoglobinuria consistent with a form of metabolic myopathy. Muscle biopsy showed only minimal alterations, whereas a marked reduction of glycosylated \u3b1-DG was evident.This case further expands the phenotypic spectrum of GMPPB mutations and highlights the importance of exhaustive molecular characterization of patients with reduced glycosylation of \u3b1-DG at muscle biopsy

A 68-year-old Caucasian man presenting with urinary bladder lymphoepithelioma: a case report

Introduction. Lymphoepithelioma is a very rare form of malignant tumor originating from epithelial line cells. Its occurrence has potential clinical, therapeutic and prognostic implications. In the present report we describe an unusual case of bladder cancer with two different histological varieties: transition cell carcinoma and lymphoepithelioma-like carcinoma. Lymphoepithelioma-like carcinoma of the bladder has only been rarely reported in the literature to date. Case presentation. We present the case of a 68-year-old Caucasian man who, after occurrence of hematuria, underwent transurethral resection of a bladder tumor. The results of a histological examination confirmed a high-grade non-muscle-invasive pT1 lymphoepithelioma-like carcinoma of the urinary bladder, associated with a concurrent high-grade transition cell carcinoma. After analyzing the histological features, our patient was subjected to treatment with intra-vesical instillations of bacillus Calmette- Guérin. Our work stresses that diagnosis and therapeutic approaches can be difficult and controversial, especially in the early stages of this rare carcinoma. Conclusions: This report emphasizes the importance of extending our knowledge and experiences regarding this uncommon carcinoma. Further studies are needed to better understand this rare disease and define more accurate diagnostic and therapeutic strategies. © 2013 Spinelli et al.; licensee BioMed Central Ltd

Pensioni e pensionati: condizioni, prospettive e sostenibilità

Nella prima parte del capitolo si descrive l’andamento dello stock e dei flussi delle pensioni nel 2020 e 2021, in aggregato, per gestione e per genere. Il tema del divario di genere è oggetto di un approfondimento che scaturisce dall’osservazione che, nel 2021, il reddito delle femmine è stato di oltre 6.000 euro inferiore a quello dei maschi. Tale differenza si ricollega al fatto che i maschi prevalgono nettamente nelle pensioni anticipate, ovvero quelle di importo più elevato, mentre le femmine hanno una netta prevalenza nelle pensioni ai superstiti e in quelle di vecchiaia. Da un confronto per età dei pensionati, emerge però che il divario in termini di frequenza delle anticipate decresce con l’età, per cui, in futuro, una quota crescente di pensionate potrebbe avere pensioni dirette di livello comparabile a quello dei maschi. Tuttavia, notevoli differenze di genere si registrano anche negli importi delle singole tipologie di pensioni. Per le prestazioni più strettamente legate all’attività lavorativa, ovvero le anticipate e la vecchiaia, il divario è in parte riconducibile alle ben note differenze in termini di continuità lavorativa, anche se il divario in termini di anzianità contributiva si è visibilmente ridotto nel tempo. Le settimane di contribuzione delle pensionate più giovani, infatti, superano dell’80% le settimane delle più anziane, ma la differenza rispetto ai maschi resta significativa. Dall’analisi dei dati emerge, inoltre, che il rapporto tra pensione lorda e settimane di contribuzione è significativamente maggiore per i maschi e questo suggerisce che il contributo alla pensione di una settimana di lavoro sia maggiore per i maschi che per le femmine. Allo scopo di misurare il potere d’acquisto dei pensionati e l’efficacia del sistema di previdenza nel fornire un reddito pensionistico in sostituzione di quello da lavoro, si è poi stimato il tasso di sostituzione delle pensioni. Tra coloro che si sono ritirati dal mercato del lavoro dopo il 2017, il tasso di sostituzione risulta pari, in media, al 75% della retribuzione massima ricevuta negli ultimi 10 anni di attività, con una differenza di 2 punti percentuali tra maschi e femmine. Rispetto agli altri paesi dell’UE, si tratta di un valore relativamente elevato. Infatti, nel confronto internazionale tra i paesi dell’UE 27, si tratta di un valore inferiore solo a quelli di Grecia, Spagna e Portogallo. La dispersione del tasso di sostituzione è però notevole, ed è in larga misura riconducibile alle differenze nell’anzianità contributiva dei pensionati. Il capitolo approfondisce poi il tema della disuguaglianza in termini di reddito pensionistico e la questione delle pensioni “povere”. Nel 2021, il 40% dei pensionati ha percepito un reddito pensionistico lordo inferiore ai 12.000 euro. Da un’analisi del ventesimo percentile di reddito pensionistico (fino a 10.000 euro nel 2021) emerge che solo il 15% dei pensionati in questa fascia riceve un assegno sociale e il 26% una pensione al superstite. Quasi il 60% percepisce una pensione di vecchiaia o anticipata dal Fondo Pensione Lavoratori Dipendenti, il che riflette l’ampiamente discusso fenomeno della cosiddetta povertà lavorativa – tema analizzato in un box che ricollega la povertà lavorativa al salario orario e ai tempi di lavoro, e anche alla composizione familiare e alla azione retributiva dello Stato. Un’analisi multivariata illustra infine come la probabilità di una pensione inferiore a 10.000 euro sia decrescente nell’anzianità contributiva, nella retribuzione, e nell’età al pensionamento (da cui dipende il tasso di trasformazione del montante contributivo in rendita pensionistica). È invece crescente nell’età di inizio lavoro (che determina la “quota contributiva” della pensione) e nella scelta di anticipare l’uscita, soprattutto per le donne (per esempio con Opzione donna che comporta il ricalcolo dell’assegno col metodo contributivo). Per quanto riguarda le differenze in termini di reddito pensionistico, nel periodo 1995-2021, l’indice di disuguaglianza di Gini dei redditi pensionistici è cresciuto di circa il 10%, attestandosi a 0,35 nel 2021, un valore inferiore a quello delle retribuzioni che è superiore a 0,4.1 La disuguaglianza è massima tra le pensioni di vecchiaia dei lavoratori dipendenti del privato (soprattutto maschi), presumibilmente per la grande variabilità della loro anzianità contributiva. Nel capitolo viene poi effettuato un confronto tra paesi per quanto riguarda la spesa pensionistica utilizzando dati Eurostat sulla base dei quali, nel 2019, l’ultimo anno per cui sono disponibili i dati riclassificati, la spesa pensionistica dell’UE 27 è stata il 12,7% del Pil. In Italia e Grecia il rapporto è stato pari al 16%, in Francia al 15%, in Germania al 12% e al 5% in Irlanda. Come prevedibile, la spesa in rapporto al PIL risulta crescente nel tasso di sostituzione della pensione rispetto all’ultimo salario, per cui nei paesi meno generosi, laddove la pensione è una frazione contenuta del salario, la spesa in rapporto al PIL è più bassa. Non emerge invece una correlazione tra la spesa media per beneficiario e il regime di calcolo della pensione. Il capitolo si chiude con sette box in cui vengono approfondite una serie di tematiche specifiche. Il primo riguarda la confluenza in INPS del Fondo di previdenza dei giornalisti INPGI/1 a partire dal 1° luglio 2022 e viene effettuato un confronto tra le pensioni dei giornalisti con quelle dei lavoratori dipendenti del settore privato e del settore pubblico. Nel secondo viene effettuato un bilancio sui pensionamenti attraverso “Quota 100”, strumento d’anticipo pensionistico, introdotto col D.L. n. 4/2019 (convertito con Legge n. 26/2019), a carattere temporaneo, valido per chi ha maturato i requisiti d’età e anzianità entro il 31 dicembre 2021. Nel terzo, viene affrontato il tema della sostenibilità del sistema pensionistico nel lungo periodo e viene stimato il risultato di esercizio del conto economico generale per tutto il periodo 2020-2029 sulla base delle previsioni dei flussi contabili di entrata e uscita di tutte le gestioni amministrate dall’Istituto. Nel quarto, vengono stimati i costi di tre possibili canali di pensionamento che aumenterebbero la flessibilità del sistema. Le proposte valutate sono l’opzione al calcolo contributivo, il calcolo della pensione con una penalizzazione che tiene conto della differenza tra età di uscita e età per la pensione di vecchiaia, e l’anticipo della quota contributiva della pensione. Nel quinto approfondimento, si affronta il tema dell’impatto della revisione dell’Irpef e dell’introduzione dell’Assegno Unico Universale (AUU) sui redditi dei pensionati italiani. Nel sesto, viene stimata la copertura assicurativa della “generazione X” (i nati tra il 1965 e il 1980), e viene valutato l’impatto di un’ipotesi di salario minimo a 9 euro sul montante contributivo di questi lavoratori. Nell’ultimo box, si affronta la questione della separazione fra spesa previdenziale e assistenziale, un tema ricorrente nel dibattito italiano in materia di welfare

Intramuscular Aripiprazole in the Acute Management of Psychomotor Agitation

Study Objective To assess acute efficacy and safety of 9.75mg of intramuscular (IM) injections of the atypical antipsychiatric aripiprazole in patients with schizophrenia or bipolar disorder and acute agitation. Design Open-label trial of IM injections of aripiprazole and 24-hour monitoring of clinical response in patients with major psychoses and acute agitation. Partial analysis of blood levels of the administered drug to correlate with clinical response. Setting Acute psychiatric care wards in a single university hospital. Patients A total of 201 acutely agitated patients (79 with schizophrenia and 122 with bipolar disorder I). Intervention Aripiprazole 9.75mg IM injection. Measurements and Main Results We evaluated clinical response using the Excitatory Component of the Positive and Negative Syndrome Scale (PANSS-EC), the Agitation/Calmness Evaluation Scale (ACES), and the Clinical Global Impressions scale (CGI). Assessments were conducted 30, 60, 90, and 120minutes and 24hours after the first injection for PANSS-EC and ACES, and 2, 4, 6, and 24hours for CGI. Response was at least a 40% decrease in PANSS-EC scores. We measured serum aripiprazole and dehydroaripiprazole levels in a subsample. IM aripiprazole significantly improved clinical measures. PANSS-EC improved progressively, starting after 30minutes. ACES improved after 90minutes and continued thereafter. Effects were sustained, with steadily decreasing CGI scores, until the 24th hour. Response rate was 83.6% after 2hours, but with repeat injections, it rose to over 90% with no differences among diagnostic groups. Although there were gender differences in the response to individual PANSS-EC items, the responses were similar overall. Neither clinical monitoring nor patient reporting revealed any side effects. No therapeutic window was identified, and levels did not correlate with any clinical measure. Conclusion Aripiprazole was effective and safe in reducing acute agitation in patients with bipolar disorder or schizophrenia. Our results compare favorably to double-blind trials, probably due to higher expectations in trials involving no placebo arm. Absence of side effects could be related to the short observation time

Ejection of damaged mitochondria and their removal by macrophages ensure efficient thermogenesis in brown adipose tissue

Recent findings have demonstrated that mitochondria can be transferred between cells to control metabolic homeostasis. Although the mitochondria of brown adipocytes comprise a large component of the cell volume and undergo reorganization to sustain thermogenesis, it remains unclear whether an intercellular mitochondrial transfer occurs in brown adipose tissue (BAT) and regulates adaptive thermogenesis. Herein, we demonstrated that thermogenically stressed brown adipocytes release extracellular vesicles (EVs) that contain oxidatively damaged mitochondrial parts to avoid failure of the thermogenic program. When re-uptaken by parental brown adipocytes, mitochondria-derived EVs reduced peroxisome proliferator-activated receptor-γ signaling and the levels of mitochondrial proteins, including UCP1. Their removal via the phagocytic activity of BAT-resident macrophages is instrumental in preserving BAT physiology. Depletion of macrophages in vivo causes the abnormal accumulation of extracellular mitochondrial vesicles in BAT, impairing the thermogenic response to cold exposure. These findings reveal a homeostatic role of tissue-resident macrophages in the mitochondrial quality control of BAT