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The study of coordination practices in health policy is a central aspect. The need for further research has been recently highlighted because of COVID-19. In this sense, dialogic practices (DP) have been identified but not validated yet. The purpose of this study is to develop and validate a DP questionnaire for healthcare teams. Items were identified based on a literature review, and the content validation was carried out by means of a Delphi study. A total of 10 experts assessed the clarity and appropriateness of the items and their corresponding measurement scales. After two rounds, a high level of consensus was reached, with agreement of 90% or higher on all items, and a high degree of stability and concordance in the results. This study resulted in a questionnaire consisting of four items, one for each identified DP initially proposed to the experts, as no other practices were revealed. From a practical perspective, the validation of these items constitutes a methodological innovation that responds to the call in the literature to open new avenues for comparative studies, and the possibility of generalising the findings and bringing together different approaches to the problem of coordination, which is key in health policy where unforeseen situations emerge.

Copy number variations (CNVs) are a type of genetic variation involving from 50 base pairs (bps) to millions of bps and, in a general point of view, can include alterations of complete chromosomes. As CNVs mean the gain or loss of DNA sequences, their detection requires specific techniques and analysis. We have developed Easy One-Step Amplification and Labeling for CNV Detection (EOSAL-CNV) by fragment analysis in a DNA sequencer. The procedure is based on a single PCR reaction for amplification and labeling of all fragments included. The protocol includes specific primers for the amplification of the regions of interest with a tail in each of the primers (one for forward and another for the reverse primers) together with primers for tail amplification. One of the primers for tail amplification is labeled with a fluorophore, allowing the amplification and labeling in the same reaction. Combination of several tail pairs and labels allows the detection of DNA fragment by different fluorophores and increases the number of fragments that can be analyzed in one reaction. PCR products can be analyzed without any purification on a DNA sequencer for fragment detection and quantification. Finally, simple and easy calculations allow the detection of fragments with deletions or extra copies. The use of EOSAL-CNV allows simplifying and reducing costs in sample analysis for CNV detection., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: The specific characteristics of copy number variations (CNVs) require specific methods of detection and characterization. We developed the Easy One-Step Amplification and Labeling procedure for CNV detection (EOSAL-CNV), a new method based on proportional amplification and labeling of amplicons in 1 PCR., Methods: We used tailed primers for specific amplification and a pair of labeling probes (only 1 labeled) for amplification and labeling of all amplicons in just 1 reaction. Products were loaded directly onto a capillary DNA sequencer for fragment sizing and quantification. Data obtained could be analyzed by Microsoft Excel spreadsheet or EOSAL-CNV analysis software. We developed the protocol using the LDLR (low density lipoprotein receptor) gene including 23 samples with 8 different CNVs. After optimizing the protocol, it was used for genes in the following multiplexes: BRCA1 (BRCA1 DNA repair associated), BRCA2 (BRCA2 DNA repair associated), CHEK2 (checkpoint kinase 2), MLH1 (mutL homolog 1) plus MSH6 (mutS homolog 6), MSH2 (mutS homolog 2) plus EPCAM (epithelial cell adhesion molecule) and chromosome 17 (especially the TP53 [tumor protein 53] gene). We compared our procedure with multiplex ligation-dependent probe amplification (MLPA)., Results: The simple procedure for CNV detection required 150 min, with <10 min of handwork. After analyzing >240 samples, EOSAL-CNV excluded the presence of CNVs in all controls, and in all cases, results were identical using MLPA and EOSAL-CNV. Analysis of the 17p region in tumor samples showed 100% similarity between fluorescent in situ hybridization and EOSAL-CNV., Conclusions: EOSAL-CNV allowed reliable, fast, easy detection and characterization of CNVs. It provides an alternative to targeted analysis methods such as MLPA., (© American Association for Clinical Chemistry 2020.)

Background: Therapy-related acute myeloid leukemia (t-AML) develops in patients with prior exposure to cytotoxic therapies. Selection of a pre-existing TP53 mutated clone prone to acquire additional mutational events has been suggested as the main pathogenic mechanism of t-AML. Here, we report a unique case of t-AML which developed from a pre-existing DNMT3A mutated clone that persisted in the patient for more than 10 years despite treatment with intensive chemotherapy and allogeneic hematopoietic stem cell transplantation (alloHSCT)., Case Presentation: A 42-year-old male was diagnosed with AML harboring a normal karyotype and mutations in the NPM1 (c.863&#95;864ins, p.W288 fs*12), DNMT3A (c.2645G > A, p.R882H), and IDH1 (c.395G > A, p.R132H) genes. He achieved complete remission with intensive chemotherapy and was subsequently submitted to alloHSCT. Eleven years later, he was given chemotherapy and radiotherapy to treat a lung carcinoma. Three years later, t-AML was diagnosed; the disease had arisen from a pre-existing DNMT3A mutated patient-origin clone that had subsequently acquired a TP53 mutation and a complex karyotype. Although a second transplantation was intended, the disease was refractory to induction chemotherapy, and the patient eventually died from disease complications. We retrospectively demonstrated the persistence and post-transplantation latency of the R882H-DNMT3A mutation using a real-time PCR allele-specific analysis at different time-points during the observation period., Discussion and Conclusion: The present case highlights the potential clinical implications of a R882H-DNMT3A mutated clone that persisted after conventional AML treatment, including alloHSCT. It also reinforces the notion of the importance of cell non-intrinsic factors, such as the hematopoietic-stress induced by chemotherapy and radiotherapy, as drivers of clonal expansion., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Objective: Hypertension-induced podocyte damage and the relationship with UAE is analyzed in diabetic and nondiabetic participants., Patients and Methods: Sixty-four hypertensive patients, 30 diabetics, with glomerular filtration rate (eGFR) greater than 60 ml/min per 1.73 m were included. Urinary albumin excretion was measured in morning urine using a nephelometric immunoassay and expressed as albumin/creatinine ratio. Urinary pellets were obtained from fresh urine and mRNA was assessed by real-time quantitative PCR. Likewise, protein podocyte-specific molecules were measured by western blot using specific antibodies., Results: Fourteen nondiabetics and 20 diabetics had increased UAE greater than 30 mg/g. In individuals with increased EUA, the mRNA expression of nephrin and CD2AP was low in diabetics, whereas only nephrin mRNA in nondiabetics. No differences were observed in podocalyxin and aquaporin-1 mRNA levels. Concerning the protein values, in both nondiabetic and diabetic patients, nephrin, CD2AP and podocalyxin were increased in patients with increased UAE, with no differences in aquaporin-1. A significant positive relationship was observed between log UAE and nephrin protein values, and an inverse association observed with mRNA., Conclusion: Hypertensive patients who had elevated UAE showed increased urinary excretion of podocyte-specific proteins coupled with a phenotype of decreased mRNA expression. The phenotype of podocyte-specific mRNA and the increment of nephrin can be used as a valuable marker of early glomerular injury.

Background: Currently, renal biopsy remains the gold standard for the diagnosis and prognosis of lupus nephritis (LN). However, it is an invasive method, and new non-invasive laboratory tests are needed to identify renal involvement without renal biopsy. Podocyte damage plays an important role in the pathogenesis and progression of systemic lupus erythematosus (SLE). We characterize whether the phenotype of urinary podocytes (viability, apoptosis, mRNA and protein levels of the podocyte-associated molecules) is a novel marker of clinical and histological features in SLE patients with or without LN., Methods: We quantified in urinary sediments of 32 SLE patients and 20 controls, mRNA and protein levels of podocalyxin, synapotopodin, podocin, nephrin and WT-1 by quantitative real-time polymerase chain reaction and western blot analysis and correlated these with clinical and histological parameters. The viability of detached urine podocytes was analysed by flow cytometry with podocalyxin and annexin V/7-AAD double staining and immunofluorescence of urine podocyte cultures., Results: The degree of a poptotic podocytes from urine samples was significantly decreased in patients with LN, especially in the active state (33% compared with 75% in controls, P < 0.001), and the majority of the detached podocytes in the urine of patients with active LN were viable (70% grew in culture). Furthermore, urinary mRNA of podocyte-associated molecules was significantly lower in patients with active LN (P < 0.05) compared with healthy controls, and protein levels of podocyte markers were significantly increased in SLE patients, especially with LN compared with SLE without LN (P < 0.05) and the healthy control group (P < 0.01). Finally, urinary protein levels of podocyte-related markers were associated with proteinuria and histological features (P < 0.05 and P < 0.01), and receiver operating characteristics curves of protein levels discriminate between LN and healthy controls with an area under the curve (AUC) between 0.91 and 0.77 (P < 0.001)., Conclusions: Urinary dedifferentiated podocytes were shown in active LN, and their protein levels correlated with proteinuria and histological features in LN. These preliminary results suggest that it could be a potentially useful non-invasive marker for evaluating the progression of glomerular disease in SLE., (© The Author 2016. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.)

Background: subclinical hyperthyroidism (SHT) is associated with harmful effects on cardiovascular system, bone metabolism and progression to clinical hyperthyroidism. Loss of weight is a common fact in patients with clinical hyperthyroidism and of particular relevance in elderly patients., Objective: to assess changes in body composition after radioiodine therapy for SHT due to toxic nodular goiter., Subjects and Methods: prospective controlled cohort study. Patients with persistent SHT due to toxic nodular goiter were purposed to receive treatment with radioiodine (treatment group) or to delay treatment until the study was over (control group). All treated patients received 555 MBq of ¹³¹I. Body composition (lean mass, fat mass and bone mineral content) was determined by dual-energy X-ray absorptiometry (DEXA) at baseline and 12 months after., Results: twenty-nine patients were studied (age 69.5 ± 11.5; 75.9% women; BMI 27.1 ± 5.7 kg/m²; serum thyrotropin (TSH) 0.20 ± 0.21 μUI/mL; serum free thyroxine (T4) 1.01 ± 0.19 ng/dL), 17 belonging to the treatment group and 12 to the control group. Study groups were comparable, although there was a trend for the treatment group to have more fat mass. No longitudinal changes in body composition were noted in either group, except for a trend to gain fat mass. However, when individuals with age > 65 years were selected, only patients who received radioiodine therapy showed a significant increase in body weight (from 64.1 ± 10.0 to 66.9 ± 9.2 kg), BMI (from 27.3 ± 4.8 to 28.7 ± 4.5 kg/m²), fat mass (from 26.1 ± 8.5 to 27.8 ± 7.9 kg), lean mass (from 36.3 ± 0.4 to 37.4 ± 0.4 kg) and skeletal muscle mass index (SMI) (from 6.0 ± 0.6 to 6.3 ± 0.6 kg/m²)., Conclusions: treatment of SHT has impact on body composition in subjects older than 65 years. Weight gain reflects increases in fat and, more interestingly, in lean mass., (Copyright AULA MEDICA EDICIONES 2014. Published by AULA MEDICA. All rights reserved.)

Nowadays, the human immunodeficiency virus infection (HIV) is a chronic disease. In the frequent clinical situations with fever, lymph nodes and loss weight it is necessary to determine their etiology, for establishing a specific treatment. Gastrointestinal opportunistic infections or gastric lymphomatous or sarcomatous process, which can accumulate Ga67, may be present in the patient with acquired immunodeficiency syndrome. We report 2 cases with gastric uptake in which endoscopy and biopsy was obtained. In the first one, with previous treatment with omeprazol and almalgate for gastroesophagic reflux, endoscopy and biopsy were normal and in the second patient an Helicobacter pylori infection was diagnosed. We think that gastric uptake of Ga67 in HIV patients, must indicate to the clinician to rule out associated pathologies.

Introduction and Objective: SPECT with 201Tl provides information regarding the degree of malignancy of cerebral tumours, their possible relapses, the differentiation of necrotic tissue in the tumours following chemotherapy or radiotherapy and permits differentiation into zones of various grades of histological malignancy., Material and Methods: We carried out a prospective analysis of the usefulness of SPECT with 201Tl for the histological prognosis of cerebral tumours. For one year 68 patients diagnosed (on CT and/or MR) as having an expansive cerebral lesion were studied. The early uptake (ICP) and retention (R) indices were calculated, and these results correlated with the morbid anatomy (AP) findings and the results obtained with surgery and stereotactic biopsy. Four patients were excluded due to lack of AP results., Results: Significant differences were found between the ICP of grade I-II astrocytomas (1.34 +/- 0.52) and glioblastomas multiformes (2.56 +/- 0.57), between the ICP of meningiomas (4.53 +/- 1.68) and metastases (2.45 +/- 0.58) and between those of meningiomas and all glial tumours. With regard to IR, we saw significant differences between the figures for meningiomas (0.63 +/- 0.13) and meningiomas with malignant relapses (0.94 +/- 0.17) and between metastases (0.8 +/- 0.03) and all glial tumours., Conclusions: From our study, it may be concluded that rapid, high captation of 201Tl (high ICP) with a slow fall (high IR), is associated with a process showing malignancy on histological study (malignant relapse of meningioma, glioblastoma multiforme, metastasis), whilst high take-up (high ICP) with rapid elimination (low IR) corresponds to a benign hypervascularized tumour (meningioma).

Twenty infertile patients with normal tubal patency were inseminated intraperitoneally (11 once, seven twice, and two three times) with spermatozoa (mean 14 x 10(6), range 0.6-48 x 10(6)) prepared by the standard swim-up technique. The occurrence of immunization to spermatozoa was looked for by the Gelatin Agglutination Test (GAT) and Tray Agglutination Test (TAT). Both tests gave negative results for all the controls (10 pregnant and 10 puerperal women). Antisperm antibodies were measured in the serum before, 30 d and 4-7 months after Intraperitoneal Insemination (IPI). The last check was done for only 14 patients, since six became pregnant as a consequence of the first treatment. Of the 14 patients studied after 4-7 months, seven had two, and two had three IPI. In the group of inseminated patients, 18 women with no basal sperm antibody did not show evidence of antibody formation after the treatment and it was not increased after insemination in the two patients who already had low antibody titre (1/32). In conclusion, despite the large number of spermatozoa inseminated and even after several IPI attempts, there was no evidence of de novo production of or increase in already present anti-sperm antibodies according to the methods used for the detection of ASA in this study.

Thoracic aspiration biopsy (TAB) constitutes a useful technique in establishing a diagnosis in diseases of the lungs and mediastinum. Results obtained from 1046 fluoroscopically-guided TABs are presented with review of the most important aspects of the technique. Diagnostic accuracy in malignancy detection was 93.8% in lung lesions (n = 984) and 74.5% in mediastinal lesions (n = 62). Sensitivity was higher in peripheral than in central lesions (96% vs. 87%, respectively). Specificity was 100% in both groups. Sensitivity in lesions smaller than 2 cm was 70% and 94% in larger lesions. Aspiration biopsies performed with Chiba and Franseen needles showed a similar sensitivity (95%) higher than with other types of needles. A pneumothorax developed in 138 patients (13.2%). Only eight of these required the use of an endothoracic tube (0.8% of all biopsies).

For couples with unexplained or male infertility, intraperitoneal (IP) insemination in induced cycles is a method that increases the chance of fertilization. Seventy-seven couples with male subfertility were subjected to 120 IP insemination cycles and 31 with unexplained sterility to 44 cycles. As a consequence of the treatment, 23 pregnancies were obtained, with pregnancy rates of 23% per cycle and 32% per patient for unexplained infertility and 11% and 17% for male subfertility. Pregnancy loss rate was quite elevated: 9 clinical abortions and 1 ectopic. Intraperitoneal insemination appears to be a relatively noninvasive way to deal with unexplained or male factor infertility, well worth trying before moving on to more invasive and costly approaches, such as gamete intrafallopian transfer or in vitro fertilization.

Direct intraperitoneal insemination (DIPI) is one of the least invasive strategies of assisted reproduction. Unexplained infertility and male sub-fertility are the major indications for DIPI. It is otherwise well known that assisted procreation gives poor results in severe male infertility. This is a report of a pregnancy that occurred as a result of a direct intraperitoneal insemination of prepared spermatozoa characterized by a particularly severe astheno-teratozoospermia in a couple unsuccessfully treated with other, more invasive methods of assisted fertilization.

The results obtained with intravenous brain angioscintigraphy (IBAS) with 99 mTc-DTPA(Sn) are evaluated in 61 patients with clinical suspicion of cerebellar, cerebral or cranial lesions of posterior location or without focal signs or with a previous IBAS in anterior view being normal or of low significance. One hundred patients without demonstrable cerebral or cranial pathology were explored to define the normal characteristics and variation of the IBAS in posteior view. Of the 61 patients with pathology, 45 showed angioscintigraphic alterations. The following angioscintigraphic patterns were found: 1. Perfusion increased, 2. Perfusion diminished, 3. Flip-flop, 4. Mixed pattern, 5. Circulatory arrest, 6. Jugular venous reflux, 7. Normal IBAS associated with an abnormal morphologicall study, 8. Normal IBAS associated with a normal morphological study.

It have been studied 90 children (obese 73, non obese or control 17), trying to analyze the following parameters: mean and standard deviation values, its circadian rhythms and relation between cortisol and insulin, correlation between both hormones and anthropometric parameters of obesity and sexual influence with all the studied parameters. The plasmatic cortisol values were higher in male, obese (180.4 +/- 11.2 ng/ml) and control (377.5 +/- 49.2 ng/ml) children. The plasmatic insulin values higher in obese children (55.3 +/- 9.5 microU/ml) verifying the hyperinsulinemia observed in obesity. There was not correlation between cortisol and insulin values with body fat. All the children showed cortisol and insulin circadian rythm. In the groups of obese children the cortisol circadian rythm was similar to the control one. However, the insulin circadian rythm was disturb. So, the acrophase was delayed two hours when male and female were analyzed all together and only one hour in the obese female group. The acrophase of cortisol and insulin rhythms in obese group is delayed in relation with the degree of obesity. The circadian rhythms of cortisol and insulin, in both obese and control groups, are not dependent. The duration of obesity do not have any relation with the mentioned alterations.

Renal uptake of 67Ga-citrate is described in a patient with biopsy-proven amyloidosis of the kidneys, due to Familiar Mediterranean Fever. After administration 150 MBq (4 mCi) 67Ga-citrate, scans were done at 48, 72, and 120 h. Intense uptake was noted in both kidneys. A renal biopsy done 5 days after the 67Ga-citrate scan revealed a pattern typical of amyloidosis. Gallium scanning can be useful in patients with fever of unknown origin. Renal amyloidosis can be considered when renal uptake of 67Ga-citrate associated with nephrotic syndrome is observed.

99MTc is an ideal nuclide for Pediatric use. Combined with DTPA is an adequate agent for renal studies. 15 patients with ages ranging from eight days to eleven years were studied (10 patients with congenital malformation and five with renal tumour). The dosimetric advantages, the obtention with a single nuclide dose of multiple images and the diagnostic usefulness in congenital malformation and renal tumour suggest their use.

Intravenous brain angioscintigraphy in combination with immediate static scan are very useful as a complement to the clinical and electroencephalographic data in diagnosis of brain death. Confusion in the differentiation of extracerebral from intracerebral flow on the dynamic radionuclide angiogram can be accomplished by identifying presence or absence of uptake in cerebral sinuses. Authors consider that the radionuclide study should be carried out in posterior view. Exploration is safe, quick to perform and easy to interpret and it enables to distinguish brain death from comas due to drug intoxication.