MicroRNAs are fine regulators of the whole-body adaptive response but their use as biomarkers is limited by the lack of standardized pre- and post-analytical procedures. This work aimed to compare different normalization approaches for RT-qPCR data analyses, in order to identify the most reliable and reproducible method to analyze circulating miRNA expression profiles in sedentary and highly-trained subjects. As the physically active status is known to affect miRNA expression, they could be effective biomarkers of the homeostatic response. Following RNA extraction from plasma, a panel of 179 miRNAs was assayed by RT-qPCR and quantified by applying different normalization strategies based on endogenous miRNAs and exogenous oligonucleotides. hsa-miR-320d was found as the most appropriate reference miRNA in reducing the technical variability among the experimental replicates and, hence, in highlighting the inter-cohorts differences. Our data showed an association between the physically active status and specific skeletal muscle- and bone-associated circulating miRNAs profiles, revealing that established epigenetic modifications affect the baseline physiological status of these tissues. Since different normalization strategies led to different outputs, in order to avoid misleading interpretation of data, we remark the importance of the accurate choice of the most reliable normalization method in every experimental setting.
Purpose The COVID-19 pandemic is importantly affecting the orthopaedic practice all over the world with Northern Italy being the first European area that faced the worst scenario. In this study, the changes in clinical practice occurred in an orthopaedic center in Milan are described. Methods Number and type of admissions, outpatients cancelled and preserved, emergency room, and intensive care unit activities have been analyzed in the timeframe of seven weeks since the beginning of the pandemic (from February 24th to April 10th) and compared with the same period in 2019. Results The planned surgical admissions declined from 2172 in 2019 to 664 in 2020 (69.42%, p < 0.0001), while emergencies increased from 158 to 268 (69.62%). The rehabilitation admissions declined from 414 to 69 (83.33%). The overall admission decreased by 63.52%, the trend showed a drop in the last weeks. Surgery performed in the COVID-19 operating room increased by 16.7% in the last week. Seven deaths occurred (0.7% of all orthopaedics and trauma admissions) compared with four (0.1%) which happened in the same period in 2019 (p = 0.004). Six of these patients were suffering from COVID-19. A total of 23,580 outpatients (93.8%) were cancelled. Emergency room consultations declined by 68.14% and 63.47% among white and green priority, respectively, while increased by 25% and 100% among yellow and red, respectively. Conclusion These numbers show the radical changed scenario in an orthopaedic center in Milan during COVID-19 pandemic. Elective surgery declined rapidly going close to zero, outpatient admissions were restricted to cases that cannot be postponed, while emergencies increased due to the role played by the hospital as referral orthopaedic centre during the pandemic. The still ongoing emergency will have important impacts on the overall orthopaedic healthcare management for the next months.
Background: The Coronavirus disease 2019 (COVID-19) outbreak has put strain on many healthcare systems around the world, with important consequences. The aim of this paper is to describe the impact of the COVID-19 pandemic on hip and knee arthroplasties in an Italian high-volume orthopedic center, located in the region of the country first and worst affected by the Coronavirus. Methods: Data from an institutional database were retrospectively analyzed to obtain the number of hip and knee arthroplasties performed from February 24 to April 10 2020. The figures were compared with those of the same 7-week period of the last year (2019). Results: The number of hip and knee arthroplasties showed a decrease from 706 in the same period of 2019 to 166 (76.5% less) in the current year. In 2019, a mean of 101 ± 9 hip and knee arthroplasties were performed per week compared with a mean of 24 ± 34 in 2020. Ten patients tested positive for SARS-CoV2 during their hospital stay. Two of these patients died after a regular postoperative period after developing unexpectedly COVID-19 during rehabilitation. The mortality in the 7-week period of the current year was 1.2% compared with 0% in 2019. Conclusion:The outbreak of COVID-19 had a considerable effect in our center on the number of hip and knee arthroplasties that rapidly decreased to 0 in parallel to the worsening of the situation in the country. Efforts will be soon requested because our practice is going to deal with the after-effects of the pandemic in the near future.
Purpose The SARS-CoV-2 outbreak affected health care systems at different levels with important consequences on health, economy, and social structures. This paper aims to analyse the impact on surgical block utilisation and efficiency in an orthopaedics reference centre in Northern Italy. Methods The timeframe chosen for the current analysis was April 2020, to be compared with the corresponding period in 2019. Number and type of procedures, first case delay, occupancy rate, and turnover time were used as indicators to benchmark the activities. Results The overall number of surgical procedures decreased by 57%, from 537 in 2019 to 230 in the same timeframe in 2020. Orthopaedic procedures predominated in 2019, with 434 cases (80.8%), while in 2020, trauma was the leading activity, with 200 cases (86.9%). Orthopaedic surgery had a relative decrement of 93% while trauma has relatively increased by 94%. Mean first case delay in orthopaedic room (OR) was two hours and 36 minutes (
Amyloid beta peptide (Aβ) causes neurodegeneration by several mechanisms including oxidative stress, which is known to induce DNA damage with the consequent activation of poly (ADP-ribose) polymerase (PARP-1). To elucidate the role of PARP-1 in the neurodegenerative process, SH-SY5Y neuroblastoma cells were treated with Aβ25–35 fragment in the presence or absence of MC2050, a new PARP-1 inhibitor. Aβ25–35 induces an enhancement of PARP activity which is prevented by cell pre-treatment with MC2050. These data were confirmed by measuring PARP-1 activity in CHO cells transfected with amylod precursor protein and in vivo in brains specimens of TgCRND8 transgenic mice overproducing the amyloid peptide. Following Aβ25–35 exposure a significant increase in intracellular ROS was observed. These data were supported by the finding that Aβ25–35 induces DNA damage which in turn activates PARP-1. Challenge with Aβ25–35 is also able to activate NF-kB via PARP-1, as demonstrated by NF-kB impairment upon MC2050 treatment. Moreover, Aβ25–35 via PARP-1 induces a significant increase in the p53 protein level and a parallel decrease in the anti-apoptotic Bcl-2 protein. These overall data support the hypothesis of PARP-1 involvment in cellular responses induced by Aβ and hence a possible rationale for the implication of PARP-1 in neurodegeneration is discussed.
(1) Background: In literature it is reported that 20–30% of psoriatic patients evolve to psoriatic arthritis over time. Currently, no specific biochemical markers can either predict progression to psoriatic arthritis or response to therapies. This study aimed to identify osteoimmunological markers applicable to clinical practice, giving a quantitative tool for evaluating pathological status and, eventually, to provide prognostic support in diagnosis. (2) Methods: Soluble (serum) bone and cartilage markers were quantified in 50 patients with only psoriasis, 50 psoriatic patients with psoriatic arthritis, and 20 healthy controls by means of multiplex and enzyme-linked immunoassays. (3) Results: Differences in the concentrations of matrix metalloproteases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), receptor activator of nuclear factor kappa-B- ligand (RANK-L), procollagen type I N propeptide (PINP), C-terminal telopeptide of type I collagen (CTx-I), dickkopf-related protein 1 (DKK1), and sclerostin (SOST) distinguished healthy controls from psoriasis and psoriatic arthritis patients. We found that MMP2, MMP12, MMP13, TIMP2, and TIMP4 distinguished psoriasis from psoriatic arthritis patients undergoing a systemic treatment, with a good diagnostic accuracy (Area under the ROC Curve (AUC) > 0.7). Then, chitinase-3-like protein 1 (CHI3L1) and MMP10 distinguished psoriasis from psoriatic arthritis not undergoing systemic therapy and, in the presence of onychopathy, MMP8 levels were higher in psoriasis than in psoriatic arthritis. However, in these latter cases, the diagnostic accuracy of the identified biomarkers was low (0.5 < AUC < 0.7). (4) Conclusions. By highlighting never exploited differences, the wide osteoimmunological biomarkers panel provides a novel clue to the development of diagnostic paths in psoriasis and psoriasis-associated arthropathic disease.
Skeletal muscle represents one of the most plastic tissues of our body thanks to the presence of heterogeneous population of myofibers that confer to skeletal muscle the functional plasticity necessary to modulate its morpho-fuctional properties in response to a wide range of external factors. Thus, alteration in fiber type composition represents a major component in muscle wasting associated with muscle diseases. Several mechanisms have been proposed to account for the alteration in the morpho-functional properties of skeletal muscle under pathological conditions. In this review we will discuss the potential catabolic mediators of muscle atrophy and wasting.
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