Search

The essay addresses the impact that digital-based organizational innovations have on the structure and function of the due diligence rule. After considering the complexity of organizational systems and the need for systematic thought control, the author relates diligence and professionalism, considering this as a synthesis of the supplier's knowledge, know-how and know-how to be, and emphasizes the renewed importance of the company's interest as a guiding criterion in the performance of the service.

Abstract The human mitochondrial DNA polymerase gamma is a holoenzyme, involved in mitochondrial DNA (mtDNA) replication and maintenance, composed of a catalytic subunit (POLG) and a dimeric accessory subunit (POLG2) conferring processivity. Mutations in POLG or POLG2 cause POLG-related diseases in humans, leading to a subset of Mendelian-inherited mitochondrial disorders characterized by mtDNA depletion (MDD) or accumulation of multiple deletions, presenting multi-organ defects and often leading to premature death at a young age. Considering the paucity of POLG2 models, we have generated a stable zebrafish polg2 mutant line (polg2 ia304 ) by CRISPR/Cas9 technology, carrying a 10-nucleotide deletion with frameshift mutation and premature stop codon. Zebrafish polg2 homozygous mutants present slower development and decreased viability compared to wild type siblings, dying before the juvenile stage. Mutants display a set of POLG-related phenotypes comparable to the symptoms of human patients affected by POLG-related diseases, including remarkable MDD, altered mitochondrial network and dynamics, and reduced mitochondrial respiration. Histological analyses detected morphological alterations in high-energy demanding tissues, along with a significant disorganization of skeletal muscle fibres. Consistent with the last finding, locomotor assays highlighted a decreased larval motility. Of note, treatment with the Clofilium tosylate drug, previously shown to be effective in POLG models, could partially rescue MDD in Polg2 mutant animals. Altogether, our results point at zebrafish as an effective model to study the etiopathology of human POLG-related disorders linked to POLG2, and a suitable platform to screen the efficacy of POLG-directed drugs in POLG2-associated forms.

Background: To tackle future health threats, the study aimed to assess healthcare workers’ (HCWs) adherence to infection prevention and control (IPC) measures and to evaluate whether knowledge and concerns related to COVID-19 could be its potential predictors. A secondary aim was to explore how and how much healthcare facilities support HCWs to implement IPC practices. Methods: This cross-sectional study was conducted between June and September 2021 in Southern Italy using a self-administered questionnaire. Socio-demographic and professional characteristics, COVID-19 preparedness and response plan in the facilities, knowledge about COVID-19’s transmission and non-pharmaceutical interventions (NPIs), concerns about the possibility of transmitting COVID-19 to family members and patients, adherence to IPC practices, and sources of information were investigated. Results: Among the 492 enrolled HCWs, the overall knowledge median score was 6 out of a maximum score of 8 and predictors of good knowledge were practicing in a facility that organized training courses about IPC measures and having more years in practice. HCWs were more concerned about the possibility of transmitting the infection to their family members (64.4%) than to negative patients (38.9%). A sizable proportion claimed to take off the face mask by touching only the straps or ties (76.3%) and to replace the face mask when it was wet (70.7%). Only 26.2% never/rarely touched the face mask while wearing it. Good adherence to preventative practices was more likely in HCWs other than physicians and practicing in a facility that organized training courses about IPC measures and in regional hospitals compared with those practicing in district hospitals. Conclusions: The study findings produce information about what worked well and where gaps were identified during COVID-19 pandemic, along with data useful for hospitals and health systems in general to better prepare and put in place appropriate interventions in case of similar future events.

Salinity stress impairs growth and physiological performance in tomato, which is one of the most economically important vegetables and is widely cultivated in arid and semi-arid areas of the world. Plant landraces, which are heterogeneous, local adaptations of domesticated species, offer a unique opportunity to valorize available germplasm, underpinning the productivity, resilience, and adaptive capacity of staple crops in vulnerable environments. Here, we investigated the response of fully mature tomato plants from a commercial variety, an ancestral wild relative, and a landrace under short-term salinity exposure, as well as their ability to recover upon cessation of stress. The heterogeneous panel evaluated in this study revealed different adaptative strategies to cope the stress. Our data highlighted the ability of the tomato clade to handle low and intermediate salinity stress for short-term exposure time, as well as its capacity to recover after the cessation of stress, although inter- and intraspecific variations in morphological and physiological responses to salinity were observed. Overall, the landrace and the wild type performed similarly to control conditions under low salinity, demonstrating an improved ability to maintain ionic balance. In contrast, the commercial genotype showed susceptibility and severe symptoms even under low salinity, with pronounced reductions in K+/Na+ ratio, PSII photochemical efficiency, and photosynthetic pigments. This research confirmed that improved salt tolerant genotypes can lead to substantial, positive impacts on horticultural production. While the salt tolerance mechanism of domesticated tomato was efficient under mild stress conditions, it failed at higher salinity levels.

Bianca Viscomi,1 Gladstone Faria,2 Claudia A Hernandez,3 Luiz Perez,4 Julieta Spada,5 Victoria De la Fuente,6 Mariana Muniz7 1Dermatologist, Bianca Viscomi Dermatologia Private Practice, São Paulo, Brazil; 2Plastic Surgeon, Gladstone Institute Private Practice, São Paulo, Brazil; 3Dermatologist, MD Dermatology Private Practice, Medellin, Colombia; 4Dermatologist, Espaço Mira Private Practice, São Paulo, Brazil; 5Dermatologist, Dermatología y Estética Private Practice, Buenos Aires, Argentina; 6Dermatologist, Senshi Derma Clinic Studio Private Practice, Mexico City, Mexico; 7Dermatologist, Mariana Muniz Dermatologia Private Practice, São Paulo, BrazilCorrespondence: Bianca Viscomi, R Correia Dias, 530 suite 131, São Paulo, SP, 04104-001, Brazil, Tel +5511 97222-0525 ; +55113051-2304, Email biancaviscomi@yahoo.com.brAbstract: A well-defined jawline is aesthetically pleasing, so that rejuvenation of the jawline is becoming part of routine aesthetic practice. Restoring balance to the lower face without surgical intervention requires a multiple treatment approach, among which the minimally invasive injectables play a central role. Nevertheless, amidst the plethora of different injectable products available, choosing the option that best suits the patient’s need can be a challenge. A panel of experts sought to describe herein 4 clinical cases, illustrating the Contouring Plus technique, which provides a practical guidance for lower third assessment, choice of products, and treatment execution, especially among hyaluronic acid and calcium hydroxylapatite, aiming for immediate, as well as mid-to-long-term results.Keywords: rejuvenation, biostimulation, neocollagenesis, collagen, calcium hydroxylapatite, hyaluronic acid, jawline treatment

The loss of nicotinamide adenine dinucleotide is reported to be associated with muscle mitochondrial dysfunction in murine cancer models. Here the authors show that niacin supplementation improves mitochondrial metabolism and reduces muscle wasting in mouse models of cachexia.

Mammalian mitochondrial respiratory chain (MRC) complexes are able to associate into quaternary structures named supercomplexes (SCs), which normally coexist with non-bound individual complexes. The functional significance of SCs has not been fully clarified and the debate has been centered on whether or not they confer catalytic advantages compared with the non-bound individual complexes. Mitochondrial respiratory chain organization does not seem to be conserved in all organisms. In fact, and differently from mammalian species, mitochondria from Drosophila melanogaster tissues are characterized by low amounts of SCs, despite the high metabolic demands and MRC activity shown by these mitochondria. Here, we show that attenuating the biogenesis of individual respiratory chain complexes was accompanied by increased formation of stable SCs, which are missing in Drosophila melanogaster in physiological conditions. This phenomenon was not accompanied by an increase in mitochondrial respiratory activity. Therefore, we conclude that SC formation is necessary to stabilize the complexes in suboptimal biogenesis conditions, but not for the enhancement of respiratory chain catalysis.

Summary: Mutations in MPV17 are a major contributor to mitochondrial DNA (mtDNA) depletion syndromes, a group of inherited genetic conditions due to mtDNA instability. To investigate the role of MPV17 in mtDNA maintenance, we generated and characterized a Drosophila melanogaster Mpv17 (dMpv17) KO model showing that the absence of dMpv17 caused profound mtDNA depletion in the fat body but not in other tissues, increased glycolytic flux and reduced lifespan in starvation. Accordingly, the expression of key genes of glycogenolysis and glycolysis was upregulated in dMpv17 KO flies. In addition, we demonstrated that dMpv17 formed a channel in planar lipid bilayers at physiological ionic conditions, and its electrophysiological hallmarks were affected by pathological mutations. Importantly, the reconstituted channel translocated uridine but not orotate across the membrane. Our results indicate that dMpv17 forms a channel involved in translocation of key metabolites and highlight the importance of dMpv17 in energy homeostasis and mitochondrial function.

This brief article argues that Italian diplomacy in the Mediterranean took an important turn around 1874. This shift is made visible through the direct intervention and discursive mobilization of Italian emigrant communities in Egypt by the Italian consul. The article specifically addresses debates at the time around legal pluralism and the importance of extraterritorial jurisdiction for diasporic national communities. In attempting to gain imperial power relative to French strongholds in the region, the young Italian state struggled to articulate a position. In this article, I show how the Italian state and its diplomatic actors used emigrant communities, and these debates, to further its aims in the years between national unification and the aggressive colonial policies enacted by the 1890s and after the turn of the century.

Abstract Background Recent clinical and experimental studies have highlighted the involvement of Ventral Tegmental Area (VTA) dopamine (DA) neurons for the early pathogenesis of Alzheimer’s Disease (AD). We have previously described a progressive and selective degeneration of these neurons in the Tg2576 mouse model of AD, long before amyloid-beta plaque formation. The degenerative process in DA neurons is associated with an autophagy flux impairment, whose rescue can prevent neuronal loss. Impairments in autophagy can be the basis for accumulation of damaged mitochondria, leading to disturbance in calcium (Ca2+) homeostasis, and to functional and structural deterioration of DA neurons. Methods In Tg2576 mice, we performed amperometric recordings of DA levels and analysis of dopaminergic fibers in the Nucleus Accumbens – a major component of the ventral striatum precociously affected in AD patients – together with retrograde tracing, to identify the most vulnerable DA neuron subpopulations in the VTA. Then, we focused on these neurons to analyze mitochondrial integrity and Apoptosis-inducing factor (AIF) localization by electron and confocal microscopy, respectively. Stereological cell count was also used to evaluate degeneration of DA neuron subpopulations containing the Ca2+-binding proteins Calbindin-D28K and Calretinin. The expression levels for these proteins were analyzed by western blot and confocal microscopy. Lastly, using electrophysiology and microfluorometry we analyzed VTA DA neuron intrinsic properties and cytosolic free Ca2+ levels. Results We found a progressive degeneration of mesolimbic DA neurons projecting to the ventral striatum, located in the paranigral nucleus and parabrachial pigmented subnucleus of the VTA. At the onset of degeneration (3 months of age), the vulnerable DA neurons in the Tg2576 accumulate damaged mitochondria, while AIF translocates from the mitochondria to the nucleus. Although we describe an age-dependent loss of the DA neurons expressing Calbindin-D28K or Calretinin, we observed that the remaining cells upregulate the levels of Ca2+-binding proteins, and the free cytosolic levels of Ca2+ in these neurons are significantly decreased. Coherently, TUNEL-stained Tg2576 DA neurons express lower levels of Calbindin-D28K when compared with non-apoptotic cells. Conclusion Overall, our results suggest that the overexpression of Ca2+-binding proteins in VTA DA neurons might be an attempt of cells to survive by increasing their ability to buffer free Ca2+. Exploring strategies to overexpress Ca2+-binding proteins could be fundamental to reduce neuronal suffering and improve cognitive and non-cognitive functions in AD.

Introduction: Biallelic variants in PITRM1 are associated with a slowly progressive syndrome characterized by intellectual disability, spinocerebellar ataxia, cognitive decline and psychosis. The pitrilysin metallopeptidase 1 (PITRM1) is a mitochondrial matrix enzyme, which digests diverse oligopeptides, including the mitochondrial targeting sequences (MTS) that are cleaved from proteins imported across the inner mitochondrial membrane by the mitochondrial processing peptidase (MPP). Mitochondrial peptidases also play a role in the maturation of Frataxin, the protein affected in Friedreich’s ataxia. Recent studies in yeast indicated that the mitochondrial matrix protease Ste23, which is a homologue of the human insulin-degrading enzyme (IDE), cooperates with Cym1 (homologue of PITRM1) to ensure the proper functioning of the preprotein processing machinery. In humans, IDE could be upregulated by Peroxisome Proliferator-Activated Receptor Gamma (PPARG) agonists.Methods: We investigated preprotein processing, mitochondrial membrane potential and MTS degradation in control and patients’ fibroblasts, and we evaluated the pharmacological effect of the PPARG agonist Pioglitazone on mitochondrial proteostasis.Results: We discovered that PITRM1 dysfunction results in the accumulation of MTS, leading to the disruption and dissipation of the mitochondrial membrane potential. This triggers a feedback inhibition of MPP activity, consequently impairing the processing and maturation of Frataxin. Furthermore, we found that the pharmacological stimulation of PPARG by Pioglitazone upregulates IDE and also PITRM1 protein levels restoring the presequence processing machinery and improving Frataxin maturation and mitochondrial function.Discussion: Our findings provide mechanistic insights and suggest a potential pharmacological strategy for this rare neurodegenerative mitochondrial disease.

Disciplining the “digital unknown” through the translation of the forms, methods and institutes of the “known analogue” can lead to significant inconsistencies and asymmetries and produce perverse effects. For this reason, this essay proposes to verify the possibility of experimenting regulatory sandboxes also in the field of labor legislation, according to what is already foreseen for some areas particularly interested in digital innovation.

Bianca Viscomi Dermatology, Private Practice, São Paulo, São Paulo, BrazilCorrespondence: Bianca Viscomi, Dermatology, Private Practice, R Correia Dias, 530 suite 131, São Paulo, São Paulo, ZIP 04104-001, Brazil, Tel +55 11 97222-0525 ; +55 113051-2304, Email biancaviscomi@yahoo.com.brAbstract: A transgender individual is someone who does not identify with his or her biological sex1. During the transgender transition process, minimally invasive procedures (MIP) provide a versatile tool in the clinician’s therapeutic armamentarium, since the changes can be impactful for the patient, yet reversible, especially for the patient who is unsure about proceeding with a definite surgery. We report 5 cases of facial feminization of transgender women, illustrating the use of minimally invasive facial remodeling procedures for male-to-female transitioning patients.Keywords: transgender, feminization, botulinum toxin, hyaluronic acid fillers, calcium hydroxyapatite, skin quality

Untreated phenylketonuria (PKU) patients and PKU animal models show hypomyelination in the central nervous system and white matter damages, which are accompanied by myelin basic protein (MBP) impairment. Despite many assumptions, the primary explanation of the mentioned cerebral outcomes remains elusive. In this study, MBP protein and mRNA expression on brains of wild type (WT) and phenylketonuric (ENU2) mice were analyzed throughout mice lifespan (14-60-180-270-360-540 post-natal days, PND). The results confirmed the low MBP expression at first PND times, while revealed an unprecedented progressive MBP protein expression recovery in aged ENU2 mice. Unexpectedly, unaltered MBP mRNA expression between WT and ENU2 was always observed. Additionally, for the same time intervals, a significant decrease of the phenylalanine concentration in the peripheral blood and brain of ENU2 mice was detected, to date, for the first time. In this scenario, a translational hindrance of MBP during initial and late cerebral development in ENU2 mice was hypothesized, leading to the execution of a microRNA microarray analysis on 60 PND brains, which was followed by a proteomic assay on 60 and 360 PND brains in order to validate in silico miRNA-target predictions. Taken together, miR-218-1-3p, miR-1231-3p and miR-217-5p were considered as the most impactful microRNAs, since a downregulation of their potential targets (MAG, CNTNAP2 and ANLN, respectively) can indirectly lead to a low MBP protein expression. These miRNAs, in addition, follow an opposite expression trend compared to MBP during adulthood, and their target proteins revealed a complete normalization in aged ENU2 mice. In conclusion, these results provide a new perspective on the PKU pathophysiology understanding and on a possible treatment, emphasizing the potential modulating role of differentially expressed microRNAs in MBP expression on PKU brains during PKU mouse lifespan.

Abstract The pathogenic mechanisms that underlie the progression of remote degeneration after spinal cord injury (SCI) are not fully understood. In this study, we examined the relationship between endoplasmic reticulum (ER) stress and macroautophagy, hereafter autophagy, and its contribution to the secondary damage and outcomes that are associated with remote degeneration after SCI. Using a rat model of spinal cord hemisection at the cervical level, we measured ER stress and autophagy markers in the axotomized neurons of the red nucleus (RN). In SCI animals, mRNA and protein levels of markers of ER stress, such as GRP78, CHOP, and GADD34, increased 1 day after the injury, peaking on Day 5. Notably, in SCI animals, the increase of ER stress markers correlated with a blockade in autophagic flux, as evidenced by the increase in microtubule-associated protein 2 light chain 3 (LC3-II) and p62/SQSTM1 (p62) and the decline in LAMP1 and LAMP2 levels. After injury, treatment with guanabenz protected neurons from UPR failure and increased lysosomes biogenesis, unblocking autophagic flux. These effects correlated with greater activation of TFEB and improved neuronal survival and functional recovery—effects that persisted after suspension of the treatment. Collectively, our results demonstrate that in remote secondary damage, impairments in autophagic flux are intertwined with ER stress, an association that contributes to the apoptotic cell death and functional damage that are observed after SCI.