Exploring capabilities of elemental mass spectrometry for determination of metal and biomolecules in extracellular vesicles.

Extracellular vesicles (EVs) are increasingly recognized as crucial components influencing various pathophysiological processes, such as cellular homeostasis, cancer progression, and neurological disease. However, the lack of standardized methods for EV isolation and classification, coupled with ambiguity in biochemical markers associated with EV subtypes, remains a major challenge. This Trends article highlights the most common approaches for EV isolation and characterization, along with recent applications of elemental mass spectrometry (MS) to analyse metals and biomolecules in EVs obtained from biofluids or in vitro cellular models. Considering the promising capabilities of elemental MS, the article also looks ahead to the potential analysis of EVs at the single-vesicle and single-cell levels using ICP-MS. These approaches may offer valuable insights into individual characteristics of EVs and their functions, contributing to a deeper understanding of their role in various biological processes.

Macrophage Migration Inhibitory Factor (MIF) is a Key Player in Dry Eye Disease.

PURPOSE: To explore the role of the proinflammatory cytokine, macrophage migration inhibitory factor (MIF), in a murine model of dry eye disease (DED). METHODS: The role of MIF on DED was determined using genetically MIF deficient mice and pharmacological inhibition of MIF. DED was induced with 0.5 mg of scopolamine via subcutaneous injection in wild type (WT) and mice lacking MIF (Mif-/-), three times a day for 21 days. DED signs, tear volume, ferning pattern and cytology impression were evaluated. Also, eye tissues were collected to determine transcripts of key inflammatory mediators and histopathological damage. In a second set of experiments, we neutralized MIF with ISO-1, an isozaxiline-derivative MIF tautomerase activity-inhibiting small molecule in WT mice, following an acute DED model for 10 days. ISO-1 was given starting on day 3 after DED induction and signs were evaluated, including a recovery phase in both experimental approaches. RESULTS: When compared to WT, Mif-/- mice showed attenuated signs of DED like preserved mucin pattern and increased tear volume. Also, Mif-/- mice maintained conjunctival epithelial cells and less corneal damage, associated with lower levels of TNFα and IL-1ß. At recovery phase, Mif-/- mice presented improved signs. Interestingly, in cornea and conjunctiva the absence of MIF selectively downregulated the transcription of inflammatory enzymes like inos and nox4 whereas displayed enhanced transcripts of il-4, il-13, tgfß and cox2. Finally, pharmacological inhibition of MIF using ISO-1, replicated the above findings in the mouse model. CONCLUSION: MIF is a central positive mediator of the inflammatory process in experimental DED, thus, targeting MIF could be used as a novel therapy in ocular surface inflammatory pathologies.

Cultural adaptation and validation of the Family Nursing Practice Scale (FNPS) for use with Spanish-speaking Nursing Degree students.

AIM: The task proposed was to conduct a cultural adaptation of the Family Nursing Practice Scale and to assess the psychometric properties of the resulting instrument. BACKGROUND: Nursing students must obtain sufficient personal competence and confidence to act with patients and their families. For this purpose, an assessment scale is needed to inform teachers of the student's progress and to determine whether further training or changes in teaching methods are required. DESIGN: A cross-sectional study design was used. METHODS: The researchers conducted this study with 202 students of nursing at two Spanish universities. In the cultural adaptation, the following steps were followed: definition of concepts, translation, back translation, expert group review and implementation by a pilot group with a subsequent cognitive interview. Internal consistency was determined by Cronbach's alpha. Reliability was verified by an initial application of the scale, followed by a repetition after seven days, analysing the results obtained in terms of the intraclass correlation coefficient, the construct validity (by Spearman's non-parametric correlation test) and confirmatory factor analysis (using JASP 0.16.4 statistical software). RESULTS: Cronbach's alpha resulted in 0.95. Spearman's correlation coefficient Family Nursing Practice Scale total with the course year was -0.26 < 0.001. The intraclass correlation coefficient for the total score of the scale was 0.91. The confirmatory factor analysis was performed on a model in which the total score for the scale was considered in relation to the two subscales, and each subscale in relation to its constituent items. The p-value associated with the chi-square was 0.550. The root mean square error of approximation and comparative fit indices presented values of <0.001 (90 % C.I. <0.001-0.048) and 1.000, respectively. CONCLUSIONS: These results indicate that the Spanish version presents good internal consistency, construct validity and reliability. Moreover, the confirmatory factor analysis confirms that it presents a good fit to the model initially proposed for this scale for Nursing Degree students.

Targeting macrophage migration inhibitory factor (MIF): a promising therapy for inflammatory ocular diseases.

Inflammatory ocular diseases are characterized by the presence of a persistent inflammatory response which cause tissue injury, decrease visual acuity and in severe cases, blindness. Several cytokines represent a therapeutic opportunity since they are key amplifiers of these pathologies, and thus neutralizing agents against them have been developed. Amongst others, macrophage migration inhibitory factor (MIF), an early produced inflammatory cytokine, has consistently been found elevated in patients with distinct ocular diseases (inflammatory and autoimmune). Here, we present and discuss evidence showing that preclinical trials using diverse strategies to neutralize MIF resulted in significant attenuation of disease signs and therefore MIF blockage might be a promising therapy for ocular diseases.

Benefits of Paediatric to Adult Transition Programme in Inflammatory Bowel Disease: The BUTTERFLY Study of GETECCU and SEGHNP.

(1) Background: Transition is a planned movement of paediatric patients to adult healthcare systems, and its implementation is not yet established in all inflammatory bowel disease (IBD) units. The aim of the study was to evaluate the impact of transition on IBD outcomes. (2) Methods: Multicentre, retrospective and observational study of IBD paediatric patients transferred to an adult IBD unit between 2017-2020. Two groups were compared: transition (≥1 joint visit involving the gastroenterologist, the paediatrician, a programme coordinator, the parents and the patient) and no-transition. Outcomes within one year after transfer were analysed. The main variable was poor clinical outcome (IBD flare, hospitalisation, surgery or any change in the treatment because of a flare). Predictive factors of poor clinical outcome were identified with multivariable analysis. (3) Results: A total of 278 patients from 34 Spanish hospitals were included. One hundred eighty-five patients (67%) from twenty-two hospitals (65%) performed a structured transition. Eighty-nine patients had poor clinical outcome at one year after transfer: 27% in the transition and 43% in the no-transition group (p = 0.005). One year after transfer, no-transition patients were more likely to have a flare (36% vs. 22%; p = 0.018) and reported more hospitalisations (10% vs. 3%; p = 0.025). The lack of transition, as well as parameters at transfer, including IBD activity, body mass index < 18.5 and corticosteroid treatment, were associated with poor clinical outcome. One patient in the transition group (0.4%) was lost to follow-up. (4) Conclusion: Transition care programmes improve patients' outcomes after the transfer from paediatric to adult IBD units. Active IBD at transfer impairs outcomes.

Risk reductions during pyrene biotransformation and mobilization in a model plant-bacteria-biochar system.

The productive application of motile microorganisms for degrading hydrophobic contaminants in soil is one of the most promising processes in modern remediation due to its sustainability and low cost. However, the incomplete biodegradation of the contaminants and the formation of the intermediary metabolites in the process may increase the toxicity in soil during bioremediation, and motile inoculants may mobilize the pollutants through biosorption. Therefore, controlling these factors should be a fundamental part of soil remediation approaches. The aim of this study was to evaluate the sources of risk associated with the cometabolism-based transformation of 14C-labeled pyrene by inoculated Pseudomonas putida G7 and identify ways to minimize risk. Our model scenario examined the increase in bioaccessibility to a distant source of contamination facilitated by sunflower (Helianthus annuus L.) roots. A biochar trap for mobilized pollutant metabolites and bacteria has also been employed. The experimental design consisted of pots filled with a layer of sand with 14C-labeled pyrene (88 mg kg-1) as a contamination focus located several centimeters from the inoculation point. Half of the pots included a biochar layer at the bottom. The pots were incubated in a greenhouse with sunflower plants and P. putida G7 bacteria. Pots with sunflower plants showed a higher biodegradation of pyrene, its mobilization as metabolites through the percolate and the roots, and bacterial mobilization toward the source of contamination, also resulting in increased pyrene transformation. In addition, the biochar layer efficiently reduced the concentrations of pyrene metabolites collected in the leachates. Therefore, the combination of plants, motile bacteria and biochar safely reduced the risk caused by the biological transformation of pyrene.

Spanish adaptation and validation of the Cultural Competence Assessment Tool (CCATool) for undergraduate nursing students.

AIM: To analyse and validate a contextually adapted version of the Papadopoulos' Cultural Competence Assessment Tool (CCATool) for Spanish undergraduate nursing students. BACKGROUND: Globalization has driven and intensified international migration. Thus, nurses must treat patients of many different cultural origins. Accordingly, both cultural competence and an appropriate tool with which to evaluate it are required. METHODS: The CCATool questionnaire was adapted for use in a Spanish context and was evaluated through a questionnaire completed by 262 undergraduate nursing students. Seven days later, the same students completed the questionnaire, without any intervention, and a subgroup of 144 students completed the questionnaire for a third time after taking a course in Transcultural Care. The reliability, sensitivity and stability of the questionnaire were assessed. RESULTS: The reliability of the questionnaire obtained an adequate Cronbach's alpha of 0.81. Stability was assessed at seven days, using the intraclass correlation coefficient, which showed good/excellent results. Moreover, the questionnaire was able to detect the changes related to the nursing educative programme, thus highlighting a good sensitivity of the tool. CONCLUSION: Cultural competence is an essential element of nurses' education, and instruction in this area should begin as soon as possible. In addition, an appropriate evaluation system is required. For this purpose, the present study describes and analyses a valid, reliable tool for use with undergraduate nursing students in Spain. IMPLICATIONS FOR NURSING AND HEALTH POLICY: Health and education policies must be sensitive to the ethnic and cultural differences of the patients who are cared for, and address the situations that cause difficulties in care. Hospitals and health centres must offer good care to patients from diverse cultures, so one of the necessary situations is the training and evaluation of the cultural competence of health professionals.

Pain control with punch grafting in ulcers with underlying arteriolosclerosis.

OBJECTIVE: Martorell hypertensive ischaemic ulcers are often misdiagnosed and can be a clinical and therapeutic challenge. Controversy exists regarding both their underlying triggers and the type of treatment that should be carried out. This study was designed to compare the effectiveness of punch grafting and conventional therapy in pain reduction. METHOD: A single-centre retrospective study was performed, including 40 patients with a clinical diagnosis of a Martorell ulcer or post-traumatic ulcer secondary to arteriolopathy in the elderly, who were treated with punch grafting (n= 24) or conventional medical treatment (n=16). RESULTS: There was a statistically and clinically significant reduction in pain after punch grafting. The minimal overall reduction was of three points in visual analogue pain scores. Of the patients who received punch grafting, 80% reported a VAS pain score of 0 at the third follow-up, in contrast with the 44% (n=4) patients who were treated without punch grafting. The mean time to epithelialisation was 82.1 days in patients who received conventional treatment and 43.5 days in those who received punch grafts. CONCLUSION: Punch grafting is a simple, validated and cost-effective technique that can be performed on an outpatient basis, promotes wound healing and reduces pain. It may control pain and stimulate epithelialisation even if the wound does not present with optimum wound bed characteristics for graft taking. Pain reduction and faster epithelialisation are associated with improvements in patients' quality of life.

Cultural adaptation and validation of the Transcultural Self-Efficacy Tool for use with undergraduate nursing students in Spain.

BACKGROUND: The provision of culturally competent care helps reduce healthcare inequalities. There is a positive association between the cultural competence of nursing professionals and patient satisfaction, and also between nurses' self-efficacy and patients' adherence to treatment. While training in this respect is important, the self-assessment of skills should also be addressed. AIMS: To produce a culturally-validated Spanish-language version of the Transcultural Self-Efficacy Tool (TSET) for use in undergraduate nursing studies, and to evaluate its implementation within the Spanish educational system. METHODS: This cultural adaptation and psychometric validation study was carried out with student nurses from two universities. International standards were followed for the translation and cultural adaptation of the questionnaire. All students completed the questionnaire twice, the second time 14 days after the first. Those who took the Transcultural Care course at the University of Málaga also completed the questionnaire a third time, after concluding this study subject. The reliability, discriminatory capacity, stability and sensitivity to change of the questionnaire were tested and confirmed, and a confirmatory factor analysis was performed. RESULTS: A total of 286 students took part in this study. Overall, the TSET obtained a reliability score of 0.978, according to Cronbach's alpha test. In relation to nationality of origin, the discriminatory capacity of the questionnaire was -1.067 (-1815 a -0,320) = 0.005. Stability, assessed at 14 days using the intraclass correlation coefficient, was 0.901 (0.873 a 0.923) <0.001. The before-after sensitivity to change for those who took the Transcultural Care course was 1.820 (1558 a 2083) <0.001. CONCLUSIONS: This Spanish-language version of the TSET is culturally suitable for the educational context of undergraduate nursing students in Spain, and its psychometric validation was performed with satisfactory results.

Anti-Phosphatidylserine/Prothrombin Antibodies in Healthy Women with Unexplained Recurrent Pregnancy Loss.

Recurrent pregnancy loss (RPL) affects up to 6% of couples. Although chromosomal aberrations of the embryos are considered the leading cause, 50% of cases remain unexplained. Antiphospholipid Syndrome is a known cause in a few cases. Antiphospholipid antibodies (aPL) anticardiolipin, anti-Beta-2-Glycoprotein-I and Lupus Anticoagulant (criteria aPL) are recommended studies in RPL workup. We tested healthy women with unexplained RPL for criteria aPL and anti-Phosphatidylserine/Prothrombin antibodies (aPS/PT). Patients were classified into three groups according to the number and pregnancy week of RPL: Extra-Criteria (EC), with 2 miscarriages, Early Miscarriage (EM), with ≥3 before pregnancy at week 10 and Fetal Loss (FL), with ≥1 fetal death from pregnancy at week 10. Circulating criteria aPL were absent in 98.1% of EM, 90.9% of FL and 96.6% of EC groups. In contrast, aPS/PT were positive in 15.4% of EM, 15.1% of FL, 16.6% of EC patients and 2.9% in controls. aPS/PT posed a risk for RPL, with an odds ratio of 5.96 (95% confidence interval (CI): 1.85-19.13. p = 0.002) for EM, 7.28 (95% CI: 2.07-25.56. p = 0.002) for FL and 6.56. (95% CI: 1.77-24.29. p = 0.004) for EC. A successful live birth was achieved in all pregnant patients positive for aPS/PT who received treatment with heparin, aspirin and/or hydroxychloroquine.

Early and Sequential Punch Grafting in the Spectrum of Arteriolopathy Ulcers in the Elderly.

INTRODUCTION: Posttraumatic ulcers secondary to age-related arteriolosclerosis may be included in the clinic-histopathological spectrum of the Martorell hypertensive ischemic ulcer. Histologically, they both present occlusive subcutaneous arteriolosclerosis. Considering these similarities, they could benefit from the same treatment. CASE REPORT: The authors present an 84-year-old white female who had a painful 9-cm x 4-cm ulcer of the central inner aspect of her left leg. The ulcer had developed 2 months prior after mild trauma, with good response to early and sequential punch grafting, combined with single-use negative pressure wound therapy and compression therapy. Pain reduction was obtained from the first punch grafting session. To achieve complete epithelialization 16 weeks after the first punch grafting procedure, 2 more sessions were necessary. CONCLUSIONS: Early punch grafting is an effective technique for pain control and healing promotion in Martorell ulcers. Punch grafting, which may be performed in an outpatient setting, is well tolerated by patients and may be repeated several times if necessary.

Punch grafting for pain reduction in hard-to-heal ulcers.

OBJECTIVE: Punch-grafting is a traditional technique to enhance wound healing, which has been associated with significant pain reduction. There are few studies measuring pain reduction after punch grafting, our study was designed to measure this outcome. METHOD: Patients with hard-to-heal wounds treated with punch grafting were included in a single centre prospective study. Wound pain intensity was measured using a Visual Analogue Scale (VAS) at baseline (before the procedure) and at three time points after the procedure. Punch grafting was performed in an outpatient setting. Patient demographic data, wound aetiology and percentage of graft take were recorded. RESULTS: A total of 136 patients were included (62 men and 74 women). Mean age was 60±35 years and 51 (38%) had venous leg ulcers (VLU), 29 (21%) had postoperative wounds, 15 (11%) Martorell ulcers, 15 (11%) traumatic wounds, four (3%) arterial ulcers and 22 (16%) 'other' ulcers. Of the patients, 38 (28%) did not present with painful ulcers and, after punch grafting, all of them remained painless; 29 (21%) patients obtained >70% pain reduction, whereas 73 (54%) patients achieved pain suppression. Pain suppression did not depend on the percentage of graft take. CONCLUSION: Punch-grafting is a simple, technique that not only promotes wound healing but also reduces pain. It can also be performed on an outpatient basis. Further studies should be performed to achieve a better understanding of this beneficial finding. Declaration of interest: The authors have no conflicts of interest to declare.

Deficiencia de vitamina D en adultos mayores con fractura de cadera / Deficiency of vitamin D in elderly adults with hip fracture

Resumen: OBJETIVO: Conocer la prevalencia de la deficiencia de vitamina D en adultos mayores con y sin fractura de cadera en un hospital de tercer nivel de atención. MATERIAL Y MÉTODO: Estudio observacional, descriptivo, retrospectivo y transversal efectuado en pacientes adultos mayores del 1 de agosto de 2016 al 1 de agosto de 2017. Para su análisis los pacientes se dividieron en dos grupos, uno con fractura de cadera y otro sin el antecedente de fractura. Se recolectaron los datos demográficos, concentraciones de 25-hidroxivitamina D [25(OH)D] y de hormona paratiroidea. La deficiencia de vitamina D se definió según la clasificación de Holick. RESULTADOS: Se incluyeron 58 pacientes; cada grupo (con y sin fractura de cadera) incluyó 29 pacientes. Las concentraciones de 25(OH)D fueron, en promedio, de 18.24 ± 7.8 ng/mL en todos los pacientes con diferencias entre los grupos de fracturados vs no fracturados: 14.83 ± 5.28 vs 21.66 ± 7.45 ng/mL, respectivamente (p < 0.001). La prevalencia de deficiencia de vitamina D fue de 63.7% y fue mayor en el grupo de fracturados en comparación con los que no tenían este antecedente: 86.2 vs 41.4%. CONCLUSIONES: La deficiencia de vitamina D es alta en la población de adultos mayores y su frecuencia se incrementa con la edad.

Abstract: OBJECTIVE: To know the prevalence of vitamin D deficiency in older adults with and without hip fracture in a third level hospital. MATERIAL AND METHOD: An observational, descriptive, retrospective and crosssectional study was conducted in elderly patients from August 1st 2016 to August 1st 2017. For their analysis, they were divided into two groups, one group with a hip fracture and the other without a fracture history. Demographic data, levels of 25-hydroxyvitamin-D [25(OH)D] and PTH were collected. Vitamin D deficiency was defined according to Holick's classification. RESULTS: There were included 58 patients, each group with 29. The levels of 25(OH) D were, on average, 18.24 ± 7.8 ng/mL in all patients, with differences between the groups of fractured vs non-fractured: 14.83 ± 5.28 ng/mL vs 21.66 ± 7.45 ng/ mL, respectively; p < 0.001. The prevalence of vitamin D deficiency was 63.7%, being higher in the fractured group compared to those without this background: 86.2% vs 41.4%. CONCLUSIONS: Deficiency of vitamin D is high in the population of elderly adults and its frequency is increased with age.

Comparative study of the efficacy and safety of bromfenac, nepafenac and diclofenac sodium for the prevention of cystoid macular edema after phacoemulsification.

AIM: To compare the efficacy, tolerability and safety of bromfenac 0.09%, nepafenac 0.1% or diclofenac 0.1% for the prophylaxis of the cystoid macular edema (CME) after phacoemulsification. METHODS: Group sequential observational comparative study. After phacoemulsification, patients received two months for topical treatment of either diclofenac sodium, bromfenac or nepafenac. All patients received concomitant topical tobramycin 0.3% and topical prednisolone 1%. We measured CME using optical coherence tomography (OCT) central foveal thickness, macular thickness and total macular volume. RESULTS: We enrolled 243 patients from January to June 2015, and 35% received diclofenac, 32.9% bromfenac and 32.1% nepafenac. When we compared pre-operative to three weeks to two months, bromfenac was more effective in reducing foveal volume (21.3 and 35.4 mm3, respectively), compared with the diclofenac (1.3 and 11.5 mm3, respectively), and the nepafenac group, became more edematous 6.4 and 5.3, respectively. Totally 133 patients completed the post-surgical satisfaction questionnaire. Patients complained of eye stickiness in 13.8% whom we gave nepafenac, versus 10.3% whom we gave diclofenac sodium, and in 0 whom we gave bromfenac. CONCLUSION: Bromfenac is the best tolerated and is more effective than diclofenac and nepafenac in reducing CME after phacoemulsification.

Martorell Hypertensive Ischemic Ulcer Successfully Treated With Punch Skin Grafting.

Martorell hypertensive ischemic ulcer can be a real clinical and therapeutic challenge. Controversy exists regarding both the underlying triggers of the disease and the type of treatment that should be established. Early skin grafting has been suggested as an effective treatment to enhance pain reduction and wound healing in these patients. The authors present the case of a 68-year-old woman with well-controlled hypertension and diabetes who developed extremely painful, rapidly progressing bilateral ulcers on the distal aspect of her legs. Without previous surgical debridement, the lesions were covered with punch grafts. Pain and necrotic progression were immediately controlled and complete epithelialization was achieved in 7 weeks.

Mouse macrophage galactose-type lectin (mMGL) is critical for host resistance against Trypanosoma cruzi infection.

The C-type lectin receptor mMGL is expressed exclusively by myeloid antigen presenting cells (APC) such as dendritic cells (DC) and macrophages (Mφ), and it mediates binding to glycoproteins carrying terminal galactose and α- or ß-N-acetylgalactosamine (Gal/GalNAc) residues. Trypanosoma cruzi (T. cruzi) expresses large amounts of mucin (TcMUC)-like glycoproteins. Here, we show by lectin-blot that galactose moieties are also expressed on the surface of T. cruzi. Male mMGL knockout (-/-) and wild-type (WT) C57BL/6 mice were infected intraperitoneally with 10(4) T. cruzi trypomastigotes (Queretaro strain). Following T. cruzi infection, mMGL-/- mice developed higher parasitemia and higher mortality rates compared with WT mice. Although hearts from T. cruzi-infected WT mice presented few amastigote nests, mMGL-/- mice displayed higher numbers of amastigote nests. Compared with WT, Mφ from mMGL-/- mice had low production of nitric oxide (NO), interleukin (IL)-12 and tumor necrosis factor (TNF)-α in response to soluble T. cruzi antigens (TcAg). Interestingly, upon in vitro T. cruzi infection, mMGL-/- Mφ expressed lower levels of MHC-II and TLR-4 and harbored higher numbers of parasites, even when mMGL-/- Mφ were previously primed with IFN-γ or LPS/IFN-γ. These data suggest that mMGL plays an important role during T. cruzi infection, is required for optimal Mφ activation, and may synergize with TLR-4-induced pathways to produce TNF-α, IL-1ß and NO during the early phase of infection.

MIF synergizes with Trypanosoma cruzi antigens to promote efficient dendritic cell maturation and IL-12 production via p38 MAPK.

Macrophage migration inhibitory factor (MIF) has been found to be involved in host resistance to several parasitic infections. To determine the mechanisms of the MIF-dependent responses to Trypanosoma cruzi, we investigated host resistance in MIF⁻/⁻ mice (on the BALB/c background) during an intraperitoneal infection. We focused on the potential involvement of MIF in dendritic cell (DC) maturation and cytokine production. Following a challenge with 5 x 10(3)T. cruzi parasites, wild type (WT) mice developed a strong IL-12 response and adequate maturation of the draining mesenteric lymph node DCs and were resistant to infection. In contrast, similarly infected MIF⁻/⁻ mice mounted a weak IL-12 response, displayed immature DCs in the early phases of infection and rapidly succumbed to T. cruzi infection. The lack of maturation and IL-12 production by the DCs in response to total T. cruzi antigen (TcAg) was confirmed by in vitro studies. These effects were reversed following treatment with recombinant MIF. Interestingly, TcAg-stimulated bone marrow-derived DCs from both WT and MIF⁻/⁻ mice had increased ERK1/2 MAPK phosphorylation. In contrast, p38 phosphorylation was only upregulated in WT DCs. Reconstitution of MIF to MIF⁻/⁻ DCs upregulated p38 phosphorylation. The MIF-p38 pathway affected MHC-II and CD86 expression as well as IL-12 production. These findings demonstrate that the MIF-induced early DC maturation and IL-12 production mediates resistance to T. cruzi infection, probably by activating the p38 pathway.