First imported case of Candida auris infection in Milan, Italy: genomic characterisation.

PURPOSE: Candida auris, an emerging multidrug-resistant yeast, has been reported worldwide. In Italy, the first case was reported in 2019. We describe the first case of C. auris, imported from Greece, in Milan, using whole genome sequencing to characterise mutations associated with antifungal resistance. CASE PRESENTATION: On October 2022 an 80-year-old Italian man was hospitalised in Greece. In the absence of clinical improvement, the patient was transferred to our hospital, in Italy, where blood culture resulted positive for C. auris. Despite therapy, the patient died of septic shock. In a phylogenetic analysis the genome was assigned to Clade I with strains from Kenya, United Arab Emirates and India. D1/D2 region resulted identical to a Greek strain, as for many other strains from different World regions, highlighting the diffusion of this strain. CONCLUSION: Importation of C. auris from abroad has been previously described. We report the first case of C. auris imported into Italy from Greece, according to phylogenetic analysis. This case reinforces the need for monitoring critically ill hospitalised patients also for fungi and addresses the need for the standardisation of susceptibility testing and strategies for diagnosis and therapy.

Chagas disease prevalence among migrants from El Salvador in Milan: a cross- sectional study of an often-overlooked population.

BACKGROUND: Chagas disease (CD) is considered to be highly endemic in El Salvador, where its prevalence is estimated to be 1.3-3.7%. Although more than 40,000 migrants from El Salvador are currently living in Europe (particularly in Spain and Italy), there are few data regarding the prevalence of CD in this population. The aim of this study was to evaluate the prevalence of CD among Salvadorans living in Italy. METHODS: A cross-sectional serological survey of CD among Salvadorans living in the metropolitan area of Milan was carried out between October 2017 and December 2019. The participants' blood samples were tested for Trypanosoma cruzi antibodies using two different serological assays. The collected demographic data included their biological sex, province of origin, the type of housing in their country of origin, and family history of CD. RESULTS: Of the 384 subjects who voluntarily participated in the study, five (1.3%, most coming from La Paz) were positive to both serological assays and therefore conclusively diagnosed as having CD. Five other subjects had discrepant serological results but were not positive to a third assay. Three of the five subjects with a diagnosis of CD completed medical staging, one of whom had chronic disease (digestive and cardiac involvement). CONCLUSIONS: The prevalence of CD among Salvadorans living in Milan is similar to that estimated by the WHO in 2010. Although they are often overlooked in CD surveys, Salvadoran migrants should be included in CD control programs in countries in which the disease is not endemic.

Prevalence and reasons for non-nursing tasks as perceived by nurses: Findings from a large cross-sectional study.

AIM: The aim of this study is to describe the prevalence and reasons for non-nursing tasks as perceived by nurses. BACKGROUND: Four types of non-nursing tasks have been identified to date: (a) auxiliary; (b) administrative, (c) expected by allied health care professionals; and (d) medical. However, no studies on a large scale have been performed with the aim of identifying the prevalence of all of these non-nursing tasks, and factors promoting or hindering their occurrence, given that they represent a clear waste of nurses' time. METHODS: A cross-sectional study in 2017, following The Strengthening the Reporting of Observational studies. All active nurses registered in an Italian provincial Nursing Board (=1331) willing to participate were involved. A questionnaire survey exploring the nature of the nursing tasks performed in daily practice and the underlying reasons was administered via paper/pencil and e-mail. RESULTS: A total of 733 nurses participated of which 94.5% performed at least one type of non-nursing task, mainly administrative and auxiliary. Auxiliary tasks are less likely among nurses working in a community (odds ratio [OR] 0.43, 95% CI 0.29-0.63, p < .01) or in a residential (OR 0.41, 95% CI 0.23-0.72, p < .01) setting, in critical (OR 0.29, 95% CI 0.16-0.54, p < .01) or surgical (OR 0.37, 95% CI 0.19-0.75, p < .01) hospital settings, and when they deal with unexpected clinical events (OR 0.58, 95% CI 0.44-0.77, p < .01). Greater adequacy of nursing resources decreases the occurrence of auxiliary tasks (OR 0.98, 95% CI 0.97-0.99, p < .01), whereas the need to compensate for a lack of resources (OR 1.44, 95% CI 1.07-1.93, p < .01) increases it. CONCLUSIONS: Around one-third of shift time is devoted to non-nursing tasks; working in a hospital, in medical units, with lack of resources and with patients with predictable clinical conditions might increase the occurrence of auxiliary tasks. IMPLICATIONS FOR NURSING MANAGEMENT: Strategies to increase the time available for nursing care should consider the type of tasks performed by nurses, their antecedents and the value added to care in terms of patient' benefits.

Non-nursing tasks as experienced by nursing students: Findings from a phenomenological interpretative study.

BACKGROUND: During their clinical learning experience, students are exposed to the nursing profession as a powerful structural reality, experiencing the so-called professional socialisation, a process recognised as the basis of professional identity. Inside this process, students progressively acknowledge their professional identity as being composed of several competencies and, among these, also non-nursing tasks. OBJECTIVES: To explore non-nursing tasks in the context of nursing students' clinical learning experiences. DESIGN: An interpretative phenomenological study design was performed and carried out in 2016. The COnsolidated criteria for REporting Qualitative (COREQ) research principles were used in reporting study methods and findings. SETTING: Two Italian Bachelor of Nursing degree programmes located in Northern Italy. PARTICIPANTS: Students attending their nursing programmes who a) had successfully passed one or more theoretical examinations; b) had one or more clinical learning experiences in varied contexts (e.g. hospital, community); c) were attending the 1st, 2nd or 3rd year, and d) were willing to participate, were interviewed with an open-ended, face-to-face, audio-recorded interview. METHODS: A thematic analysis was performed. RESULTS: Participating students (n = 18) were between 20 and 25 years old and were attending the 1st to the 3rd (and final) academic year. Non-nursing tasks were experienced by them according to three main themes: a) "Being out of the scope of the learning experience," b) "Being forced by external and internal forces," and c) "Dealing with mixed outcomes by looking for a compromise." All students have reported learning to perform non-nursing tasks by shadowing clinical nurses and also practising these tasks by themselves. Internal and external forces prompted students to perform non-nursing tasks, which were recognised as having positive, negative, and neutral effects on themselves and on their learning outcomes. CONCLUSIONS: Non-nursing tasks are acquired since the beginning of the clinical experience, thus shaping the nursing students' professional identity. At the undergraduate nursing level, strategies should be implemented to prevent the phenomena that a) threaten the acquisition of more complex nursing competences expected by patients and society, and b) shape future generations to be flexible and to perform different tasks, included those below their role.

Nursing students' involvement in shift-to-shift handovers: Findings from a national study.

BACKGROUND: Effective performance of clinical handovers should be one of the priorities of nursing education to promote efficient communication skills and ensure patient safety. However, to date, no studies have explored to what extent nursing students are involved in handovers. OBJECTIVE: To explore nursing students' handover involvement during their clinical rotations and associated factors. METHOD: This was a secondary analysis of a large national cross-sectional study that involved 9607 undergraduate nursing students in 27 universities across 95 three-year Italian baccalaureate nursing programs. The involvement in the clinical handovers was the end point (from 0, never, to 3, always). A path analysis was performed to identify variables directly and indirectly affecting students' handover involvement. RESULTS: Handover involvement was reported as 'only a little', 'to some extent', and 'always' by 1739 (18.1%), 2939 (30.6%), and 4180 (43.5%) students, respectively; only 749 (7.8%) of students reported never being involved. At the path analysis explaining the 19.1% of variance of nursing students' involvement, some variables emerged that directly increased the likelihood of being involved in handovers. These were being female (ß = 0.115, p < 0.001); having children (ß = 0.107, p = 0.011); being a 3rd-year student (ß = 0.142, p < 0.001) and being a 2nd-year student as compared to a 1st-year student (ß = 0.050, p = 0.036); and having a longer clinical rotation (ß = 0.015, p < 0.001) in units with high 'quality of the learning environment' (ß = 0.279, p < 0.001). Moreover, students who were supervised by the nurse teacher (ß = -0.279, p < 0.001), or by a nurse on a daily basis (ß = -0.253, p = 0.004), or by the staff (ß = -0.190, p < 0.001) reported being less involved in handovers as compared to those students supervised by a clinical nurse. Variables with indirect effects also emerged (model of student's supervision adopted at the unit level, and number of previous clinical rotations attended by students). Moreover, handover involvement explained 11.5% of students self-reported degree of competences learned during the clinical experience. CONCLUSIONS: Limiting students' opportunity to be involved in handover can prevent the development of communication skills and the professional socialization processes. Strategies at different levels are needed to promote handover among undergraduate nursing students.

Opportunity to discuss ethical issues during clinical learning experience.

BACKGROUND: Undergraduate nursing students have been documented to experience ethical distress during their clinical training and felt poorly supported in discussing the ethical issues they encountered. RESEARCH AIMS: This study was aimed at exploring nursing students' perceived opportunity to discuss ethical issues that emerged during their clinical learning experience and associated factors. RESEARCH DESIGN: An Italian national cross-sectional study design was performed in 2015-2016. Participants were invited to answer a questionnaire composed of four sections regarding: (1) socio-demographic data, (2) previous clinical learning experiences, (3) current clinical learning experience quality and outcomes, and (4) the opportunity to discuss ethical issues with nurses in the last clinical learning experience (from 0 - 'never' to 3 - 'very much'). PARTICIPANTS AND RESEARCH CONTEXT: Participants were 9607 undergraduate nursing students who were attending 95 different three-year Italian baccalaureate nursing programmes, located at 27 universities in 15 Italian regions. ETHICAL CONSIDERATIONS: This study was conducted in accordance with the Human Subject Research Ethics Committee guidelines after the research protocol was approved by an ethics committee. FINDINGS: Overall, 4707 (49%) perceived to have discussed ethical issues 'much' or 'very much'; among the remaining, 3683 (38.3%) and 1217 (12.7%) students reported the perception of having discussed, respectively, 'enough' or 'never' ethical issues emerged in the clinical practice. At the multivariate logistic regression analysis explaining 38.1% of the overall variance, the factors promoting ethical discussion were mainly set at the clinical learning environment levels (i.e. increased learning opportunities, self-directed learning, safety and nursing care quality, quality of the tutorial strategies, competences learned and supervision by a clinical nurse). In contrast, being male was associated with a perception of less opportunity to discuss ethical issues. CONCLUSION: Nursing faculties should assess the clinical environment prerequisites of the settings as a context of student experience before deciding on their accreditation. Moreover, the nursing faculty and nurse managers should also enhance competence with regard to discussing ethical issues with students among clinical nurses by identifying factors that hinder this learning opportunity in daily practice.

Multilevel National Analysis of Nursing Students' Perceived Opportunity to Access Evidence-Based Tools During Their Clinical Learning Experience.

BACKGROUND: Easy-to-access tools have been demonstrated to improve evidence-based practice (EBP) competences among nursing students. However, students' perception of access to EBP tools (e.g., clinical practice guidelines [CPGs], protocols) is unknown. AIMS: To explore: (1) nursing students' opportunity to access EBP tools during their education, and (2) associated factors. METHODS: A national cross-sectional study including all Italian nursing programs. Nursing students were deemed eligible according to the following inclusion criteria: Those who (1) were attending or just ended their practical rotation lasting at least 2 weeks at the time of the survey, and (2) expressed through written informed consent their willingness to take part in the study. Participants were asked about their perceived opportunity to access EBP tools during their most recent clinical learning experience (from 0 - not at all to 3 - always). A set of explanatory variables was collected at the individual, nursing program, and regional levels by using a questionnaire. RESULTS: Nine thousand six hundred and seven (91.6%) out of 10,480 nursing students took part in the study. Overall, 4,376 (45.6%) students perceived not at all or only a small opportunity to access EBP tools during their most recent clinical rotation. In the multilevel analysis, factors promoting access were mainly set at the clinical learning environment level (high safety and nursing care quality, high self-directed learning opportunities, high quality of the learning environment, and being supervised by a clinical nurse). In contrast, male gender and lower academic class were associated with a lower perception of accessibility to EBP tools. A consistent variability in the perceived opportunity to access EBP tools emerged across regions. LINKING EVIDENCE TO ACTION: Evidence-based decision-making is increasingly expected from nurses. Therefore, nursing faculties should safeguard and continuously improve students' competence regarding EBP, by implementing strategies mainly at the nursing program and regional levels.

[The quality of life as perceived by older people. Results of a phenomenological study]. / La qualità di vita raccontata dagli anziani. Risultati di una ricerca fenomenologica.

INTRODUCTION: Elderly Quality of life (QoL) is vulnerable because of the decline in physical and mental capacity, discharge from work, rupture of the family and isolation. Many QoL Scales are made for adults: there is little research investigating how older people perceive QOL. AIM: This study aims to explore and understand the perceptions that older people have about their QOL. METHOD: A qualitative research hermeneutics-phenomenological was done. Narratives were analyzed by mixed method phenomenological-grounded (Mortari, 2007). RESULTS: 16 elderly were interviewed (aged between 80 and 93 years), residents in their home. The analysis of the interviews confirm that QoL is a multidimensional concept that encompasses several components of life. There were 7 shared themes, reflecting the experience of well-being in older people living at home. The dimensions are: living relationships that drive away loneliness, fill the time with activities that follow their own interests, accept themselves as elderly person, choose how and where to live, think about death. CONCLUSIONS: The results of the study show that older people perceive QOL as made of multiple components and variables. Elderly are focused on the remaining capacities, on the acceptance of the positive aspects of their lives rather than on what they can no longer do. This study help to identify some social and health strategies to guarantee the best way to live the last part of life.

[Back to units for nursing students' education? The Dedicated Education Units (DEU)]. / Un ritorno ai "reparti scuola"? Le unità dedicate alla formazione (DEU).

. Back to units for nursing students' education? The Dedicated Education Units (DEU). INTRODUCTION: The reorganization and rationalization of resources and cost containment in health care put a strain on the sustainability of practical training of student nurses. The Dedicated Education Units (DEU), where ward staff, in collaboration with university teachers, receive large numbers of students, integrating the caring and teaching missions, are a possible answer. AIM: To describe the main characteristics of DEUs. METHODS: A literature search was conducted in Pubmed with the following key-words Dedicated Education Unit, Education Unit and Nursing Education, up to January 30, 2017. RESULTS: Several models of DEU were identified with differences in contexts, professional roles involved, type of organizations (number of students, length of practical training). The students perceive a welcoming climate that promotes learning and allows time and space for reflection; they develop a professional group identity and learn to recognize and implement the presponsibilities related to the professional role. The students express satisfaction for the relationship with professionals involved in their education due to the clear definition of roles and responsibilities, of their learning needs and feel supported in the connections of theory and practice. The DEU, receiving large number of students optimize the use of resources. CONCLUSIONS: The DEU represent one of the possible models of organization of the practical training, able to ensure a high quality learning environment. The practical implications of its implementation in the italian context on skills acquisition and sustainability need a thorough assessment.

[The Italian instrument evaluating the nursing students clinical learning quality]. / Lo strumento italiano di misurazione della qualitàdell'apprendimento clinico degli studenti infermieri.

. The Clinical Learning Quality Evaluation Index for nursing students. INTRODUCTION: The Italian nursing programs, the need to introduce tools evaluating the quality of the clinical learning as perceived by nursing students. Several tools already exist, however, several limitations suggesting the need to develop a new tool. AIM: A national project aimed at developing and validating a new instrument capable of measuring the clinical learning quality as experience by nursing students. METHODS: A validation study design was undertaken from 2015 to 2016. All nursing national programs (n=43) were invited to participate by including all nursing students attending regularly their clinical learning. The tool developed based upon a) literature, b) validated tools already established among other healthcare professionals, and c) consensus expressed by experts and nursing students, was administered to the eligible students. RESULTS: 9606 nursing in 27 universities (62.8%) participated. The psychometric properties of the new instrument ranged from good to excellent. According to the findings, the tool consists in 22 items and five factors: a) quality of the tutorial strategies, b) learning opportunities; c) safety and nursing care quality; d) self-direct learning; e) quality of the learning environment. CONCLUSIONS: The tool is already used. Its systematic adoption may support comparison among settings and across different programs; moreover, the tool may also support in accrediting new settings as well as in measuring the effects of strategies aimed at improving the quality of the clinical learning.

Inhibidores adquiridos de la coagulación: enfoque diagnóstico y casos especiales / Acquired inhibitors of blood coagulation: diagnostic perpective and especial cases / Inibidores adquiridos da coagulação: abordagem diagnóstica e casos especiais

Los inhibidores adquiridos son defectos raros. Se asocian a diferentes manifestaciones clínicas con morbimortalidad significativa. Su detección es importante para implementar el tratamiento sin demora. Hay inhibidores específicos (bloquean función), contra todos los factores de la coagulación y además VWF, o de interferencia (afectan una o varias vías de la coagulación). Los inhibidores específicos son alo-anticuerpos desarrollados en pacientes deficitarios (complicación terapéutica) o auto-anticuerpos presentes en individuos sin alteraciones previas. Hay anticuerpos específicos que pueden afectar la depuración pero no inhiben función. Inhibidores de interferencia: inmunoglobulinas u otras sustancias (heparina/heparinoides, PDF/pdf, PIVKAS, moléculas anómalas, etc.) asociadas a diferentes situaciones clínicas (asintomáticos, sangrados, trombosis y/o complicaciones obstétricas). El laboratorio es fundamental para el diagnóstico. Las pruebas globales detectan el defecto, que no corrigen por el agregado de plasma normal; se caracteriza luego el tipo de inhibidor y eventualmente se titula. Esto es complejo; hay variabilidad en los resultados y posibilidad de falsos positivos o negativos, además las pruebas no son estrictamente específicas. Los algoritmos diagnósticos son útiles, pero no contemplan la posibilidad de defectos combinados. Es crítico caracterizar al inhibidor y descartar posibles interferencias o defectos concomitantes; ello requiere aplicar las pruebas adecuadas e interpretarlas correctamente.

Acquired inhibitors are rare disorders. They are associated with different clinical behaviours and significant morbi-mortality. Detection is important in order to start treatment urgently. There are either specific inhibitors, which block function, against all coagulation factors, and VWF, or with interference effects, on one or more coagulation pathways. Specific inhibitors are either allo-antibodies developed in deficient patients, which give rise to therapeutic complication; or auto-antibodies, which are present in individuals without previous defects. There are specific antibodies that can affect clearance but which cannot block the function. Inhibitors with interference effects are immunoglobulins or other substances (heparin/heparinoids, FDP/fdp, PIVKAS, abnormal molecules, etc.) associated with different clinical settings (asymptomatic, bleeding, thrombosis and/or obstetric complications). Laboratory results are fundamental for the diagnosis. Global tests are able to detect the defect, which is not corrected by the addition of normal plasma; the type of inhibitor is then characterized and titration of the inhibitor is eventually performed. This is complex; there is variability in the results and there is likelihood of false positive or negative results; moreover, the tests are not strictly specific. Diagnostic algorithms are useful tools but they do not consider combined defects. It is a critical point to characterize the inhibitor and exclude possible interferences or concomitant defects; this demands application of the correct tests without misinterpretations.

Os inibidores adquiridos são defeitos raros. Associam-se a diferentes manifestações clínicas com morbimortalidade significativa. Sua detecção é importante para implementar o tratamento sem demora. Há inibidores específicos (bloqueiam função), contra todos os fatores da coagulação e também VWF, ou de interferência (afetam uma ou várias vias da coagulação). Inibidores específicos: - aloanticorpos desenvolvidos em pacientes deficitários (complicação terapêutica); - autoanticorpos, em indivíduos sem alterações prévias. Existem anticorpos específicos que podem afetar a depuração, mas não inibem função. Inibidores de interferência: imunoglobulinas ou outras substâncias (heparina/heparinoides, PDF/pdf, PIVKAS, moléculas anômalas, etc.) associadas a diferentes situações clínicas (assintomáticos, sangramentos, tromboses e/ou complicações obstétricas). O laboratório é fundamental para o diagnóstico. Os testes globais detectam o defeito, que não corrige pelo acréscimo de plasma normal; caracteriza-se depois o tipo de inibidor e eventualmente é titulado. Isto é complexo; há variabilidade nos resultados e possibilidade de falsos positivos ou negativos, além disso os testes não são rigorosamente específicos. Os algoritmos diagnósticos são úteis, mas não consideram a possibilidade de defeitos combinados. É crítico caracterizar o inibidor e descartar possíveis interferências ou defeitos concomitantes; isso requer aplicar os testes adequados e interpretá-los corretamente.

[Validation of the Italian Clinical Learning Environment Instrument (SVIAT):study protocol]. / Strumento di Valutazione Italiano degli Ambienti di Tirocinio per gli studenti infermieri (SVIAT): protocollo di validazione.

UNLABELLED: . Validation of the Italian Clinical Learning Environment Instrument (SVIAT): study protocol. INTRODUCTION: Nursing students obtain most of their university credits in internship environments whose quality can affect their clinical learning. Several tools are available to measure the quality of the clinical learning environment (CLE) as perceived by students: these instruments developed in other countries, were validated in Italian but do not discriminate those CLEs capable (or not) to promote significant clinical learning. AIM: To validate an instrument to measure the capability of the CLE to generate clinical learning; the secondary aim is to describe the learning environments as perceived by nursing students according to individual course site and tutorial models adopted. METHODS: The study will be developed in three phases: a) instrument development and pilot phase, b) validation of the psychometric properties of the instrument and c) description of the CLEs as perceived by the students including factors/item confirmed in the validation process. Expected outcomes. A large validation, with more than 8,000 participating students is expected; the construct under lying will be confirmed through exploratory and confirmatory factor analysis and will report high internal consistency; the instrument will report also a high test-retest and inter-rater reliability; in addition, the instrument will demonstrate predictive ability by discriminating those units able (or not) to activate effective learning processes.

Inhibidor lúpico en situaciones clínicas diferentes al síndrome antifosfolipídico / Lupus inhibitor in clinical situations other than antiphospholipid syndrome

El inhibidor lúpico (IL) es uno de los criterios de laboratorio para Síndrome Antifosfolipídico (SAF); sin embargo, puede detectarse en individuos asintomáticos o estar asociado a otras situaciones clínicas. Presentamos un análisis retrospectivo de 1000 exámenes consecutivos para IL (TTPA, DRVVT) de los cuales 249 casos no presentaban criterios clínicos de SAF. Aplicando los criterios SSC-ISTH, hallamos IL+ en 27,30% (205/751) y 43,37% (108/249) de los casos con y sin criterios clínicos de SAF respectivamente; analizándose en estos últimos casos las características clínicas y de laboratorio. Contexto clínico de casos IL+ sin SAF: 18,52% asintomáticos, 34,26% síntomas de sangrado y 47,22% otras manifestaciones. Otras alteraciones de laboratorio en casos IL+ sin SAF, con síntomas de sangrado: detectamos alteraciones plaquetarias, descenso de VWF:RCo y/o VWF:Ag, disminución de FVIII, FV, FVII, FXI o fibrinógeno e hiperfibrinolisis en el 54,05% de los casos. El análisis mostró detección de IL+ en un número importante de estudios (108/1000) sin criterios SAF. Los casos con IL+ y sangrado representan un desafío particular, al requerir evaluar otros posibles defectos subyacentes, que pudiesen justificar el comportamiento clínico. La detección e identificación de defectos combinados requiriere de un análisis minucioso, a fin de alcanzar un diagnóstico correcto, esencial para tomar decisiones terapéuticas adecuadas. (AU)

Despite lupus anticoagulant (LA) is one of the laboratory criteria for antiphospholipid syndrome (APS), it can be present in asymptomatic subjects or it can be associated with other clinical settings. We present a retrospective analysis of 1000 consecutive LA assays (APTT, DRVVT), 249 of them were performed in patients without clinical criteria for APS. According to ISTH criteria, positive LA was found in 27.30% (205/751) and 43.37% (108/249) of cases with or without APS criteria respectively; in the last group, the analysis of clinical background and laboratory characteristics was done. Clinical background of LA+ cases without APS: 18.52% asymptomatic, 34.26% bleeding symptoms and 47.22% other clinical settings. Other abnormal laboratory tests in LA+ cases without APS and bleeding symptoms: platelet dysfunction; low VWF:RCo and/or VWF:Ag; decrease of FVIII, FV, FVII, FXI or fibrinogen and hyperfibrinolysis were found in the 54.05% of the cases. The analysis showed positive LA in an important number of cases (108/1000) without criteria of APS. Those LA+ cases with bleeding symptoms represent a particular challenge because other possible underlying defects have to be analysed in order to explain the clinical behaviour. The detection and identifications of combined defects required a careful analysis in order to achieve accurate diagnosis, essential for therapeutic decisions. (AU)

Identificación por nuestro grupo de una nueva mutación en el gen GP1BA (P.W246L) asociada al fenotipo PT-VWD. Sexta mutación reportada internacionalmente / Identification by our group of a novel mutation in the GP1BA gene (P.W246L) responsible for PT-VWD. The 6th mutation internationally reported

La enfermedad de von Willebrand tipo plaquetario (PT-VWD) y tipo 2B (2B-VWD) son trastornos hemorrágicos raros, caracterizados por agregación plaquetaria a bajas concentraciones de ristocetina (RIPA). El diagnóstico diferencial no es fácil y representa un desafío. Hasta el presente, sólo se habían reportado cinco mutaciones en el gen GP1BA relacionadas con este desorden. Describimos aquí la sexta mutación relacionada con PT-VWD, en un paciente con sintomatología hemorrágica severa, macro-trombocitopenia, leve agregación plaquetaria espontánea, RIPA positivo a 0,3 y 0,4 mg/mL, VWF:RCo/VWF: Ag<0,2 y estudios discriminatorios positivos para PT-VWD. VWFpp/VWF: Ag resultó normal a diferencia del 2B-VWD que en algunas oportunidades resulta afectado. El exón 28 del gen VWF del paciente y su madre no reveló mutaciones. Identificamos una sustitución G>T en el nucleótido 3805 en el gen GP1BA del paciente, resultando en un cambio de Trp a Leu en el residuo 246 (p.W246L), en la región de la GPIBa que une al VWF. Esta mutación no se identificó en su madre ni en 100 controles sanos. Es considerada como dañina por análisis in sílico. Consideramos que esta sustitución es responsable del fenotipo PT-VWD del paciente. Dada la ausencia de la misma en los 100 normales estudiados, no se considera un polimorfismo

Platelet-type von Willebrand disease (PT-VWD) and type 2B von Willebrand disease (2B-VWD) are rare bleeding disorders characterized by increased ristocetin-induced platelet aggregation (RIPA) at low concentrations. Diagnosis of either condition is not easy and the differential diagnosis is especially challenging. Five mutations in the GP1BA gene related to PT-VWD and near 50 patients are currently reported worldwide. We herein describe a patient with severe bleeding symptoms, macro thrombocytopenia, mild spontaneous platelet aggregation, positive RIPA at 0.3 and 0.4 mg/mL, VWF: RCo/VWF: Ag <0.2, normal VWFpp/VWF: Ag ratio, and RIPA mixing tests and cryoprecipitate challenge positive for PT-VWD. GP1BA gene was studied in the patient, his mother, and 100 healthy control subjects. We identified a substitution G>T at nucleotide 3805 in the patient's GP1BA gene, resulting in a Trp to Leu amino acid change at residue 246 (p.W246L), within the VWF binding region. This mutation was absent in his unaffected mother and also in the 100 controls, and was predicted as damaging by in silico analysis. The residue is located in a strongly conserved position in the phylogenetic tree. These findings argue in favor of considering this substitution does not represent a polymorphism, and is therefore responsible for the PT-VWD phenotype of the patient

Identification of p.W246L as a novel mutation in the GP1BA gene responsible for platelet-type von Willebrand disease.

Platelet-type von Willebrand disease (PT-VWD) and type 2B von Willebrand disease (2B-VWD) are rare bleeding disorders characterized by increased ristocetin-induced platelet aggregation (RIPA) at low concentrations of ristocetin. Diagnosis of either condition is not easy and the differential diagnosis between the two entities is especially challenging as evidenced by high levels of misdiagnosis of both conditions, but particularly PT-VWD. Five mutations in the GP1BA gene related to PT-VWD and less than 50 patients are currently reported worldwide. We herein describe a patient with severe bleeding symptoms, macrothrombocytopenia, mild spontaneous platelet aggregation, positive RIPA at 0.3 and 0.4 mg/mL, von Willebrand factor ristocetin cofactor (VWF:RCo) to antigen (VWF:Ag) < 0.2, normal VWF propeptide/VWF:Ag ratio, and RIPA mixing tests and cryoprecipitate challenge positive for PT-VWD. GP1BA gene was studied in the patient, in his mother, and in 100 healthy control subjects. We identified a heterozygous substitution G > T located at nucleotide 3805 in the g.DNA of the patient's GP1BA gene, resulting in a Trp to Leu amino acid change at residue 246 (p.W246L). This mutation was absent in his unaffected mother and also in the 100 controls, and was predicted as damaging by in silico analysis. The residue W246 is located within the VWF-binding region and exists in a strongly conserved position in the phylogenetic tree, which is expected to be unable to tolerate substitutions without changing its functional characteristics. These findings argue strongly in favor of the view that this substitution does not represent a polymorphism and is therefore responsible for the PT-VWD phenotype of the patient.

Morphological distribution of µ chains and cd15 receptors in colorectal polyp and adenocarcinoma specimens.

BACKGROUND: We have recently investigated the localisation of immunoglobulin-producing cells (IPCs) in inflamed intestinal tissue samples from patients with inflammatory bowel disease (IBD), and identified two main patterns of B lymphocyte infiltration: one characterised by the moderate strong stromal localisation of small B1 cell-like IgM+/CD79+/CD20-/CD21-/CD23-/CD5 ± IPCs, and the other by the peri-glandular localisation of IPCs with irregular nuclei that had surface markers specific for a B cell subset (IgM and CD79), but quantitative differences in their λ and κ chains. The same patients were also tested for CD15+ receptors, which were localised on inflammatory cell surfaces or in the crypts of the intestinal epithelium. CD15+ receptor distribution in inflamed tissues was limited to the cell structures. The aim of the study was to analyse variations in IPCs and CD15+ cell morphology or distribution in bowel biopsy specimens taken from patients with pre-malignant polyps or adenocarcinomas. METHODS: IPCs were analysed by means of immunofluorescence using polyclonal goat anti-human µ chains. The pre-malignant polyp specimens were tested for B cell surface phenotype λ and κ chains, CD79, CD20, CD21 and CD23 using an immunoperoxidase method. CD15+ cells were evaluated using the immunoperoxidase method and monoclonal anti-CD15 IgM. RESULTS: The study involved 14 patients (four with pre-malignant polyps and 10 with colorectal adenocarcinomas). The distribution of µ chains and CD15 markers varied in all of the biopsies, but delineated normal cell structures in the pre-malignant polyp specimens. B cell surface phenotype analysis of µ chain-positive cells identified a subset of CD79+/CD20-/CD21-/CD23- IPCs. The IPCs in certain areas showed the sporadic disintegration of inflammatory cell membranes or the accumulation of fluorescence in individual cells. IPC membrane disintegration was particularly marked in all of the adenocarcinoma samples, in which the CD15 markers also showed epithelial cell involvement. Furthermore, six of the ten adenocarcinoma samples had atypical and reorganised membranes that expressed an excess of both receptors and isolated small portions of tissue within the tumour. CONCLUSION: The findings of this preliminary morphological study suggest the presence of membrane disintegration and remodelling mechanisms in the tumours. The newly-formed membranes expressed high concentrations of inflammatory cell receptors that can confer adhesive properties.

Unusual B cell morphology in inflammatory bowel disease.

B lymphocytes express various different types of surface immunoglobulins that are largely unrelated to other hematological lines, although some reports have described a relationship between malignant B cells and other cells such as macrophages. Multiple genes of hematopoietic lineage, including transcription factors, are co-expressed in hematopoietic stem cells and progenitors, a phenomenon referred to as "lineage priming". Changes in the expression levels and timing of transcription factors can induce the lineage conversion of committed cells, which indicates that the regulation of transcription factors might be particularly critical for maintaining hierarchical hematopoietic development. The aim of this study was to evaluate the surface markers of particular IgM-positive and irregularly nucleated cells detected in patients with inflammatory bowel disease (IBD), and to assess their association with diagnosis and inflammatory cell recruitment. Small intestine, colon and rectal biopsy specimens of 96 IBD patients were studied. Immunoglobulin-producing cells (IPCs) were analyzed by means of immunofluorescence using polyclonal rabbit anti-human Ig and goat anti-human IgM. The specimens positive for B cells with irregular nuclei were assessed using monoclonal antibodies specific for CD79, and λ and κ chains in order to confirm their B cell nature. CD15+ cells, an important marker of inflammatory cell recruitment, were also evaluated. Statistical correlations were sought between the histological findings and clinical expression. 34 (35.4%) of the 96 patients (64 with ulcerative colitis and 32 with Crohn's disease) presented a periglandial localization of IPCs with irregular nuclei, which showed surface markers specific for the B cell subset, such as IgM and CD79, but quantitative differences in λ and κ chains. These specimens also contained CD15-positive cells, which are usually absent in healthy controls. The quantitative aspects and localization of the CD15-positive cells correlated with the distribution of the IPCs with irregular nuclei. IPCs with irregular nuclei were significantly more frequent in those patients with Crohn's disease than in those with ulcerative colitis (p<0.001). The finding of a subpopulation of cells that simultaneously showed irregular nuclei and B cell markers, such as functional surface IgM, in patients with IBD suggests that an unusual subgroup of B cells that correlates with CD15 expression and a diagnosis of Crohn's disease may be observed in the inflammatory process.

Vitamina K epóxido reductada (VKORC1): polimorfismos y sensibilidad a los dicumarínicos / Vitamin K epoxide reductase (VKORC1): polimorphisms and sensitivity to coumarin

Introducción: Los antagonistas de la vitamina K, tienen como blanco el complejo vitamina K epóxido reductasa. Hay polimorfismos (SNPs) en el gen de la subunidad 1 (VKORC1), como 1173 C>T (rs9934438) (intrón 1) y -1639 G>A (rs9923231) (región 3'UTR), asociados con diferente sensibilidad a los dicumarínicos. Objetivo: confirmar la relación entre estos SNPs y la dosis media requerida para una correcta anticoagulación. Material y Método: Estudiamos 102 pacientes (15-87 años) bajo tratamiento anticoagulante oral crónico, principalmente acenocumarol. La genotipificación fue realizada usando RFLPs (amplificación del ADN por PCR, seguida por digestión con Msp I para -1639 G>A y Sty I para 1173 C>T); analizándose luego la dosis media en los portadores de los diferentes SNPs. La frecuencia alélica obtenida para 1173 C>T fue similar a la calculada para -1639 G>A, observándose que 1179 C>T y -1639 G>A se hallan en desequilibrio de unión. La dosis media fue más alta en los pacientes portadores hemocigotos del alelo 1173 CC o -1639 GG que en los no portadores. La dosis media fue menor en individuos mayores de 70 años, independientemente de su genotipo. Reultados: Estos confirman la relación entre los SNPs analizados y la dosis de dicumarínicos.

Vitamin K epoxide reductase complex is the target of Vitamin K antagonists. Polymorphism (SNPs) in the gene of subunit 1 (VKORC1), 1173 C>T (rs9934438) (intrón 1) y -1639 G>A (rs9923231) (3'UTR), are associated with different sensitivity to oral anticoagulants. Objectives: our aim was to comfirm the relationship between these SNPs and the mean dose required for anticoagulation. Material and methods: One hundred and two patients (15-87 years) on chronic oral anticoagulant therapy, mainly acenocoumarol, were tested. Genotyping was performed using RFLPs (DNA's PCR amplification, followed by digestion with Msp I to Sty I to 1173 C>T). The mean dose of anticoagulantin carriers of different SNPs was calculated. A similar allelic frequency was obtained for 1173 C>T and -1639 G>A with linkage desequilibrium between them. The mean dose was higher in patients homozygous for 1173 CC or -1639 GG alleles than in non carriers. In those individuals over the age of 70, regardless of genotype, a lower mean dose was observed. Results: The results confirm the relationship between SNPs and oral anticoagulants.