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Epitope tags recognized by specific antibodies have been widely used over the last few decades, notably to localize tagged proteins within cells by immunofluorescence. The diversity of tags and antibodies usually prevents a side‐by‐side comparison of the efficiency with which each antibody recognizes its cognate tag. We expressed chimeric proteins, each composed of an invariant domain (IL2Ra) associated with a specific epitope tag. Double immunofluorescence allowed us to quantify in parallel the reference signal generated by the anti‐IL2Ra antibody and the signal generated by the anti‐epitope tag antibody. Since all antibodies used in this study were recombinant antibodies fused to the same mouse Fc domain, the generated signals were directly comparable. Three groups of tags/antibodies were revealed: ‘good’ antibodies generated high signals even when used at a low concentration (50 ng·mL−1), ‘fair’ antibodies generated a high signal only at high concentrations (5000 ng·mL−1), and ‘mediocre’ antibodies generated positive but weak signals. Except for an anti‐myc antibody, similar results were obtained when cells were fixed in paraformaldehyde or methanol. These results provide a side‐by‐side quantitative evaluation of different tag/antibody pairs. This information will be useful to optimize the choice of epitope tags and to choose optimal antibodies.

Abstract Background Rotator Cuff Related Shoulder Pain (RCRSP) is the most common type of shoulder pain and the most disabling common symptom in people with Rotator Cuff Tear (RCT). The patient’s point of view concerning health status has become important in decision-making procedures and has therefore been considered a possible criterion standard for assessing treatment efficacy. The study aims to explore patients’ experiences and perceptions during pre-admission for Rotator Cuff Repair surgery. Methods A qualitative descriptive study was conducted using a phenomenological approach based on Husserl’s philosophical perspective. A consecutive sample of twenty RCT patients awaiting repair surgery agreed to participate and was interviewed until the information was saturated. None of the patients enrolled was lost during the data collection phases. Data were collected through open-ended interviews between December 2021 and January 2022. The credibility, reliability, confirmability, and transferability criteria of Lincoln and Guba have been adopted to guarantee the trustworthiness of the results. The data analysis was conducted according to inductive content analysis. Results Four main themes and sub-themes related to each have been identified from the phenomenological analysis. The major themes were: (1) Pain changes lifestyle habits, (2) Pain control requires specific strategies, (3) Suffering turns time into waiting, (4) Waiting for Surgery between trust and fear. Conclusion Investigating patients’ experiences and the emotional impact of Rotator Cuff Tear facilitates the development of specific educational and therapeutic strategies to improve care and post-intervention outcomes.

Abstract Introduction Chronic pain is a distressing condition that should be treated in specialized pain clinics. Pain clinics offer a holistic, evidence-based approach, including pharmacological, complementary, and invasive treatments. This study aimed to provide preliminary information regarding chronic pain treatments and identify reasons for accessing an important hub-spoke pain clinic network. Methods A retrospective multicenter cross-sectional study was carried out. A total of 1606 patients’ records were included. Patients were selected from the 26 pain clinics of a single region in Italy. Univariate and multivariate logistic regression models were used. Results Multivariate models showed that the use of opioids were considered effective for severe or moderate pain [odds ratio (OR) 0.41; 95% 0.33–0.51], while the use of invasive treatments (OR 2.45; 95% 1.95–3.06) and the use of complementary therapy (OR 1.87; 95% 1.38–2.51) were associated with severe or moderate pain. Overall, age, sex, nonsteroidal anti-inflammatory drugs (NSAID) use, a combination of NSAIDs, complementary therapies, and a combination of opioids and invasive treatments did not seem to be significantly associated with the nature of pain. Multivariate models confirmed that clinical parameters such as the nature of pain, multi-diagnosis, more than one site of pain, treatments, and general practitioner, but not the severity of pain and use of invasive treatments, had an impact on the choice of a pain clinic. Conclusion Opioids are useful in managing moderate or severe chronic pain. Multimodal approaches are used for the management of chronic pain. Moreover, it is not clear how patients are addressed to access different pain clinics (spoke versus hub) networks. More widespread adoption is needed for an interdisciplinary approach to managing chronic pain and adopting guideline recommendations, and rigorous research is required to provide more substantial evidence and support clinical practice.

Despite process similarities, distinctive manufacturing technologies offer hyaluronic acid dermal fillers with different in vitro physicochemical and rheological properties due to peculiar crosslinked hydrogel networks. A better understanding of dermal filler properties could provide specific clinical indications and expectations with more accurate performance correlations. In this study, with an emphasis on the degree of modification, hyaluronic acid concentration and molecular weight, these process parameters were able to modulate dermal filler properties, especially rheology. Moreover, an extensive characterization of commercial hyaluronic acid injectables of the Hyal System line was described to present product properties and help to elucidate related clinical effects. Standardized methodologies were applied to correlate in vitro parameters with feasible clinical indications. In view of an optimized dermal filler design, the results of the extrudability measurements allowed the quantification of the effect of hydrogel composition, rheological properties and needle size on injectability. Composition, dynamic viscosity and needle size showed an impactful influence on hydrogel extrudability. Finally, the positive influence of 200 KDa hyaluronic acid in comparison to fragments of ether-crosslinked hyaluronic acid on fibroblast recognition were shown with a migration assay.

IntroductionThe licensure exam in nursing has always focused on the curricula used in universities. ‘Tuning’ was the first project that sought to harmonize training purposes regarding competences and learning outcomes in Europe. The Tuning educational structures have been offered in various disciplines, including nursing with the development of the Tuning Nursing Project.The study describes which of 47 Tuning Nursing Competences were evaluated during the licensure exam in nursing degree courses, and what types of trials were used for their assessment.MethodsA multicentric observational study was conducted in 4 universities in Italy. Data were collected in academic years 2017–2019, using two grids: one for cognitive and one for psychomotor tests.ResultsThe Tuning competences were requested 7522 times. The most frequently demanded were those associated with domain number two, “Nursing practice and clinical decision making”. The level of performance most required in cognitive tests was the autonomy of judgement, and both tests concerned the fields of non-communicable diseases and the hospitalized adult patient.ConclusionsThe competences most often assessed coincided with those deemed core for the first cycle of studies at the European level. Unfortunately, it has been detected a high degree of discrepancy in the types of tests used in different schools.

Abstract Background Cisd1 and Cisd2 proteins share very similar structures with an N-terminal membrane-anchoring domain and a C-terminal cytosolic domain containing an iron-cluster binding domain and ending with a C-terminal KKxx sequence. Despite sharing a similar structure, Cisd1 and Cisd2 are anchored to different compartments: mitochondria for Cisd1 and endoplasmic reticulum for Cisd2. The aim of this study was to identify the protein motifs targeting Cisd2 to the ER and ensuring its retention in this compartment. Results We used new recombinant antibodies to localize Cisd1 and Cisd2 proteins, as well as various protein chimeras. Cisd2 is targeted to the ER by its N-terminal sequence. It is then retained in the ER by the combined action of a C-terminal COPI-binding KKxx ER retrieval motif, and of an ER-targeting transmembrane domain. As previously reported for Cisd1, Cisd2 can alter the morphology of the compartment in which it accumulates. Conclusion Although they share a very similar structure, Cisd1 and Cisd2 use largely different intracellular targeting motifs to reach their target compartment (mitochondria and endoplasmic reticulum, respectively).

Since the pandemic began nurses were at the forefront of the crisis, assisting countless COVID-19 patients, facing unpreparedness, social and family isolation, and lack of protective equipment. Of all health professionals, nurses were those most frequently infected. Research on healthcare professionals' experience of the pandemic and how it may have influenced their life and work is sparse. No study has focused on the experiences of nurses who contracted COVID-19 and afterwards returned to caring for patients with COVID-19. The purpose of this study was therefore to explore the lived personal and professional experiences of such nurses, and to describe the impact it had on their ways of approaching patients, caring for them, and practicing their profession. A phenomenological study was conducted with 54 nurses, through 20 individual interviews and 4 focus groups. The main finding is that the nurses who contracted COVID-19 became “wounded healers”: they survived and recovered, but remained “wounded” by the experience, and returned to caring for patients as “healers,” with increased compassion and attention to basic needs. Through this life-changing experience they strengthened their ability to build therapeutic relationships with patients and re-discovered fundamental values of nursing. These are some of the ways in which nurses can express most profoundly the ethics of work done well.

The COVID-19 emergency has led many health facilities to reorganize themselves in a very short time to meet the urgent needs for intensive, semi-intensive or ordinary care of SARS-CoV-2 patients. In this pandemic, characterized by speed of transmission and severity of respiratory symptoms, care has been affected by the increase in volume and clinical complexity of patients, the sudden and unpredictable staff decrease and the lack of support from family members / caregivers. At the same time, experience in the field has shown how “informal” resources have been activated, which enabled to treat the highest possible number of patients above the real availability of resources. The purpose of this study was to explore the experiences of nurses involved in frontline care (COVID Centers) during the pandemic with a particular focus on professional motivation and on the development of technical-professional and personal skills. A study with a qualitative research design using focus group technique was conducted. Two focus groups were held with nine nurses. Data were analyzed with inductive content analysis. The findings can be summarized in five main categories: professional identity; motivation and sense of mission; development of professional and personal skills; spirituality; person-centered care; uniqueness of the lived experience. These findings shed new light on the correlation between motivation, professional identity and value, sense of duty and sense of belonging to the professional group. Moreover, the experience in the COVID Centers represented a valuable opportunity for participants to rediscover some specific issues related to nursing professional identity and to develop new personal and technical-professional skills in a very short time. Finally, nurses experienced once again how the nurse-patient relationship and basic care are essential to provide effective and excellent care, even and especially for patients in critical conditions. Nurses re-discovered, in a careful body care and basic care, irreplaceable elements to give back to patients, often dying, their own dignity, and all the needed closeness and attention necessary also to compensate the absence of the loved ones. These elements represent a way to concretely and deeply express the ethics of a job well done in nursing.

Sarcopenia is a progressive and generalized skeletal muscle disorder associated with poor outcomes and complications, including falls, fractures, physical disability, and death. The aim of this review is to assess the possible influence of sarcopenia on outcomes of sarcopenia in patients who underwent knee or hip replacement. A systematic review was performed using the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. Medline, EMBASE, Scopus, CINAHL, and CENTRAL bibliographic databases were searched. General study characteristics extracted were: primary author and country, year of publication, type of study, level of evidence (LOE), sample size, mean age, gender, follow-up, type of surgery, diagnosis, and outcomes. At the final screening, five articles met the selection criteria and were included in the review. Sarcopenia influences the Barthel Index (BI), which is significantly lower compared to patients without sarcopenia, which indicates that the patient is subjected to a worsening of this condition that can influence their normal life since they will become dependent on someone else. No difference in mortality rate was found was found between the studies. This systematic review addressed the possible role of sarcopenia in patients undergoing joint replacement surgery. Despite the lack of high-quality literature on this topic, a general trend in considering sarcopenia as a negative factor for quality of life in joint replacement patients was reported. However, the lack of significant results means it is not possible to report useful conclusions.

Abstract Background One of the most common bacteria responsible for most Periprosthetic joint infection (PJI) is Propionibacterium acnes. Even though the rate of infections in patients undergoing total shoulder arthroplasty is increasing, effective diagnostic tests and the precautions taken during the surgery are not yet adequate. This systematic review aims to evaluate the effectiveness of antimicrobial prophylaxis in PJI in shoulder replacement and to provide health workers with the best approach to the use of antimicrobial agents based on currently available clinical evidence. Methods a systematic review of the literature was carried out in accordance with the PRISMA Statement. Studies concerning the effectiveness of antimicrobial prophylaxis in the prevention of PJI in patients undergoing shoulder replacement were included. Results Seven studies were included in the final analysis because they were considered valid. A total of 3272 patients underwent a surgical procedure, most of which were males. The male population has a greater presence of hair, therefore a greater risk of P. acnes. in surface cultures. Patients were assessed at an average follow-up period of 20 months ranging from 9 weeks to 53 months. Conclusion The optimal perioperative antimicrobial regimen is controversial. The clinical guidelines recommend the use of only one antibiotic as prophylaxis but considering the increase in the rates of antibiotic-resistant infections, the question arises whether antibiotic prophylaxis should be extended for adequate coverage. Shoulder arthroplasty performed on the male population must be carefully checked after surgery for the possible presence of P. Acnes.

LrrkA is a Dictyostelium discoideum kinase with leucine-rich repeats. LrrkA stimulates Kil2 and intra-phagosomal killing of ingested bacteria in response to folate. In this study, we show that genetic inactivation of lrrkA also causes a previously unnoticed phenotype: lrrkA KO cells exhibit enhanced phagocytosis and cell motility compared to parental cells. This phenotype is cell autonomous, is reversible upon re-expression of LrrkA, and is not due to an abnormal response to inhibitory quorum-sensing factors secreted by D. discoideum in its medium. In addition, folate increases motility in parental D. discoideum cells, but not in lrrkA KO cells, suggesting that LrrkA plays a pivotal role in the cellular response to folate. On the contrary, lrrkA KO cells regulate gene transcription in response to folate in a manner indistinguishable from parental cells. Overall, based on analysis of mutant phenotypes, we identify gene products that participate in the control of intracellular killing, cell motility, and gene transcription in response to folate. These observations reveal a mechanism by which D. discoideum encountering bacterially-secreted folate can migrate, engulf, and kill bacteria more efficiently.

Phagocytic cells ingest and kill bacteria, a process essential for the defense of the human body against infections. Many potential killing mechanisms have been identified in phagocytic cells, including free radicals, toxic ions, enzymes, and permeabilizing peptides.

Background: The application of virtual and augmented reality technologies to orthopaedic surgery training and practice aims to increase the safety and accuracy of procedures and reducing complications and costs. The purpose of this systematic review is to summarise the present literature on this topic while providing a detailed analysis of current flaws and benefits. Methods: A comprehensive search on the PubMed, Cochrane, CINAHL, and Embase database was conducted from inception to February 2021. The Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines were used to improve the reporting of the review. The Cochrane Risk of Bias Tool and the Methodological Index for Non-Randomized Studies (MINORS) was used to assess the quality and potential bias of the included randomized and non-randomized control trials, respectively. Results: Virtual reality has been proven revolutionary for both resident training and preoperative planning. Thanks to augmented reality, orthopaedic surgeons could carry out procedures faster and more accurately, improving overall safety. Artificial intelligence (AI) is a promising technology with limitless potential, but, nowadays, its use in orthopaedic surgery is limited to preoperative diagnosis. Conclusions: Extended reality technologies have the potential to reform orthopaedic training and practice, providing an opportunity for unidirectional growth towards a patient-centred approach.

Background and Objectives: Unicompartmental knee arthroplasty (UKA) is a valid alternative to total knee arthroplasties (TKAs) in selected cases. After surgery, patients’ experience and satisfaction were traditionally evaluated by pre- and postsurgical scores and Patient-Reported Outcome Measures (PROMs). Otherwise, a statistically significant change does not necessarily correlate to a clinically meaningful improvement when measured using PROMs. To evaluate the real effect of a specific treatment and understand the difference between groups in a clinical trial, it is necessary to use a meaningful quantum of change on the score assessed. The minimal clinically important difference (MCID) and the Patient Acceptable Symptom State (PASS) can provide this meaningful change. This paper aimed to calculate the MCID and the PASS of the Forgotten Joint Score (FJS-12) after UKA. Materials and Methods: A total of 40 patients with a mean age 72.5 ± 6.4 years undergoing UKA were assessed preoperatively and six months postsurgery using the FJS-12 and the Oxford Knee Score (OKS). The baseline and 6-month postoperative scores were compared using the Wilcoxon signed ranks test. The correlation was calculated with Spearman’s rho. Both distribution-based approaches and anchor approaches were used to estimate MCID for the FJS-12. The 75th percentile and the Receiver operating characteristic (ROC) curve methods were used to calculate the PASS of FJS-12. Results: MCID estimates for normalized FJS-12 for UKA ranged from 5.68 to 19.82. The threshold of the FJS-12 with ROC method was 72.92 (AUC = 0.76). The cut-off value computed with the 75th percentile approach was 92.71. Conclusions: The MCID and PASS represent valid tools to assess the real perception of clinical improvement in patients who underwent UKA. The MCID value of FJS-12 was 12.5 for patients who underwent UKA. The value of the PASS for the FJS-12 in patients who underwent UKA was 72.92.

This systematic review aimed to assess the advantages of biosensors in detecting biomarkers for the early diagnosis of osteoarthritis (OA). OA is the most prevalent musculoskeletal disease and is a leading cause of disability and pain worldwide. The diagnosis of OA could be performed through clinical examinations and imaging only during the late stages of the disease. Biomarkers could be used for the diagnosis of the disease in the very early stages. Biosensors could detect biomarkers with high accuracy and low costs. This paper focuses on the biosensors mainly adopted to detect OA markers (electrochemical, optical, Quartz crystal microbalance, molecular and wearable biosensors). A comprehensive search on PubMed, Cochrane, CINAHL and Embase databases was conducted from the inception to November 2020. The Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines were used to improve the reporting of the review. The Methodological Index for Non-Randomized Studies (MINORS) was used for quality assessment. From a total amount of 1086 studies identified, only 19 articles were eligible for this study. The main advantages of the biosensors reported were accuracy, limited cost and ease of use, compared to traditional methods (ELISA). Otherwise, due to the lack of data and the low level of evidence of the papers included, it was impossible to find significant results. Therefore, further high-quality studies are required.

Background and Objectives: Sleep disorders are one of the most common complaints of patients with rotator cuff (RC) tears. However, potential correlations between the treatment of RC tears and the causal factors of sleep disorders are still under discussion. The aim of this review is to evaluate quality of sleep in patients before and after surgery for RC tears and to identify which factors affected patients’ sleep. Materials and Methods: A systematic review was conducted. To provide high quality of the review, the included studies were evaluated with the standardized tool “Quality Assessment Tool for Quantitative Studies” developed by the Effective Public Health Practice Project. Results: The search strategy yielded 78 articles. After duplicate removal and titles, abstracts and full-texts review, four studies were included in the systematic review. Concerning shoulder function, the most frequently reported scale was the Simple Shoulder Test (SST). Regarding sleep quality, the most frequently reported score was the Pittsburgh Sleep Quality Index (PSQI). Conclusion: We found that the majority of patients with RC tears had a sleep disturbance, especially before surgery with a general improvement in sleep quality post-operatively. Moreover, sleep quality was correlated with pain and it also seems that factors as comorbidities, obligatory position during night time, preoperative and prolonged postoperative use of narcotics and psychiatric issues may play an important role in sleep quality.

Dictyostelium discoideum is a widely used model to study molecular mechanisms controlling cell adhesion, cell spreading on a surface, and phagocytosis. In this study we isolated and characterize a new mutant created by insertion of a mutagenic vector in the heretofore uncharacterized spdA gene. SpdA-ins mutant cells produce an altered, slightly shortened version of the SpdA protein. They spread more efficiently than WT cells when allowed to adhere to a glass substrate, and phagocytose particles more efficiently. On the contrary, a functional spdA knockout mutant where a large segment of the gene was deleted phagocytosed less efficiently and spread less efficiently on a substrate. These phenotypes were highly dependent on the cellular density, and were most visible at high cell densities, where secreted quorum-sensing factors inhibiting cell motility, spreading and phagocytosis are most active. These results identify the involvement of SpdA in the control of cell spreading and phagocytosis. The underlying molecular mechanisms, as well as the exact link between SpdA and cell spreading, remain to be established.

Dictyostelium discoideum has largely been used to study phagocytosis and intracellular killing of bacteria. Previous studies have shown that Phg1A, Kil1 and Kil2 proteins are necessary for efficient intracellular killing of Klebsiella bacteria. Here we show that in phg1a KO cells, cellular levels of lysosomal glycosidases and lysozyme are decreased, and lysosomal pH is increased. Surprisingly, overexpression of Kil1 restores efficient killing in phg1a KO cells without correcting these lysosomal anomalies. Conversely, kil1 KO cells are defective for killing, but their enzymatic content and lysosomal pH are indistinguishable from WT cells. The killing defect of phg1a KO cells can be accounted for by the observation that in these cells the stability and the cellular amount of Kil1 are markedly reduced. Since Kil1 is the only sulfotransferase characterized in Dictyostelium, an (unidentified) sulfated factor, defective in both phg1a and kil1 KO cells, may play a key role in intracellular killing of Klebsiella bacteria. In addition, Phg1B plays a redundant role with Phg1A in controlling cellular amounts of Kil1 and intracellular killing. Finally, cellular levels of Kil1 are unaffected in kil2 KO cells, and Kil1 overexpression does not correct the killing defect of kil2 KO cells, suggesting that Kil2 plays a distinct role in intracellular killing.

Besides its role in controlling the morphology of mitochondria, mitofusin-2 has been proposed to tether mitochondria to the endoplasmic reticulum (ER), based largely on light microscopic analysis. In this study we have examined by electron microscopy the organization of ER and mitochondria in cells expressing or not mitofusin-2. Contrary to previous studies, we observed that loss of mitofusin-2 increased ER-mitochondria juxtaposition. These results suggest that mitofusin-2 does not play a critical role in the juxtapostion of ER and mitochondria, and highlight the essential role of ultrastructural analysis to visualize and measure contact between two intracellular compartments.