Validation of serum ferritin values by magnetic susceptometry in predicting iron overload in dialysis patients

Validation of serum ferritin values by magnetic susceptometry in predicting iron overload in dialysis patients.BackgroundGuidelines for treating anemia in dialysis patients accept, as high-end range of serum ferritin useful to optimize erythropoietin therapy, values high as 500 to 900 μg/L, on the hypothesis that ferritin might be not representative of iron overload.MethodsA superconducting quantum interference device (SQUID) was used to make direct noninvasive magnetic measurements of nonheme hepatic iron content in 40 dialysis patients treated with intravenous iron, and liver iron content was compared with biochemical markers of iron status.ResultsOnly 12/40 (30%) patients showed normal hepatic iron content (SQUID <400 μg/g), while 32.5% had mild (400 to 1000 μg/g) and 37.5% severe (>1000 μg/g) iron overload, although 28/40 patients (70%) had serum ferritin below 500 μg/L. Among many parameters, hepatic iron content was only correlated with ferritin (r = 0.324, P = 0.04). The receiver operating characteristic (ROC) analysis showed the best specificity/sensitivity ratio to identify iron overload for ferritin >340 μg/L (W = 0.716). Multivariate logistic regression analysis demonstrated that an increase in serum ferritin of 100 μg/L and female gender were independent variables associated with moderate to severe hepatic iron overload: OR 1.71 (95% CI 1.10 to 2.67) and OR 10.68 (95% CI 1.81 to 63.15), respectively.ConclusionHepatic iron overload is frequent in dialysis patients with ferritin below currently proposed high-end ranges, and the diagnostic power of ferritin in indicating true iron stores is better than presumed. Safety concerns should prompt a reevaluation of acceptable iron parameters, focusing on potential gender-specific differences, to avoid potentially harmful iron overload in a majority of dialysis patients, mainly females

Unsupervised neural networks as a support tool for pathology diagnosis in MALDI-MSI experiments:A case study on thyroid biopsies

Artificial intelligence is getting a foothold in medicine for disease screening and diagnosis. While typical machine learning methods require large labeled datasets for training and validation, their application is limited in clinical fields since ground truth information can hardly be obtained on a sizeable cohort of patients. Unsupervised neural networks - such as Self-Organizing Maps (SOMs) - represent an alternative approach to identifying hidden patterns in biomedical data. Here we investigate the feasibility of SOMs for the identification of malignant and non-malignant regions in liquid biopsies of thyroid nodules, on a patient-specific basis. MALDI-ToF (Matrix Assisted Laser Desorption Ionization -Time of Flight) mass spectrometry-imaging (MSI) was used to measure the spectral profile of bioptic samples. SOMs were then applied for the analysis of MALDI-MSI data of individual patients' samples, also testing various pre-processing and agglomerative clustering methods to investigate their impact on SOMs' discrimination efficacy. The final clustering was compared against the sample's probability to be malignant, hyperplastic or related to Hashimoto thyroiditis as quantified by multinomial regression with LASSO. Our results show that SOMs are effective in separating the areas of a sample containing benign cells from those containing malignant cells. Moreover, they allow to overlap the different areas of cytological glass slides with the corresponding proteomic profile image, and inspect the specific weight of every cellular component in bioptic samples. We envision that this approach could represent an effective means to assist pathologists in diagnostic tasks, avoiding the need to manually annotate cytological images and the effort in creating labeled datasets

Case Report: Rare Homozygous RNASEH1 Mutations Associated With Adult-Onset Mitochondrial Encephalomyopathy and Multiple Mitochondrial DNA Deletions

Mitochondrial DNA (mtDNA) maintenance disorders embrace a broad range of clinical syndromes distinguished by the evidence of mtDNA depletion and/or deletions in affected tissues. Among the nuclear genes associated with mtDNA maintenance disorders, RNASEH1 mutations produce a homogeneous phenotype, with progressive external ophthalmoplegia (PEO), ptosis, limb weakness, cerebellar ataxia, and dysphagia. The encoded enzyme, ribonuclease H1, is involved in mtDNA replication, whose impairment leads to an increase in replication intermediates resulting from mtDNA replication slowdown. Here, we describe two unrelated Italian probands (Patient 1 and Patient 2) affected by chronic PEO, ptosis, and muscle weakness. Cerebellar features and severe dysphagia requiring enteral feeding were observed in one patient. In both cases, muscle biopsy revealed diffuse mitochondrial abnormalities and multiple mtDNA deletions. A targeted next-generation sequencing analysis revealed the homozygous RNASEH1 mutations c.129-3C&gt;G and c.424G&gt;A in patients 1 and 2, respectively. The c.129-3C&gt;G substitution has never been described as disease-related and resulted in the loss of exon 2 in Patient 1 muscle RNASEH1 transcript. Overall, we recommend implementing the use of high-throughput sequencing approaches in the clinical setting to reach genetic diagnosis in case of suspected presentations with impaired mtDNA homeostasis

α-Synuclein seeding activity in duodenum biopsies from Parkinson's disease patients

Abnormal deposition of α-synuclein is a key feature and biomarker of Parkinson's disease. α-Synuclein aggregates can propagate themselves by a prion-like seeding-based mechanism within and between tissues and are hypothesized to move between the intestine and brain. α-Synuclein RT-QuIC seed amplification assays have detected Parkinson's-associated α-synuclein in multiple biospecimens including post-mortem colon samples. Here we show intra vitam detection of seeds in duodenum biopsies from 22/23 Parkinson's patients, but not in 6 healthy controls by RT-QuICR. In contrast, no tau seeding activity was detected in any of the biopsies. Our seed amplifications provide evidence that the upper intestine contains a form(s) of α-synuclein with self-propagating activity. The diagnostic sensitivity and specificity for PD in this biopsy panel were 95.7% and 100% respectively. End-point dilution analysis indicated up to 106 SD50 seeding units per mg of tissue with positivity in two contemporaneous biopsies from individual patients suggesting widespread distribution within the superior and descending parts of duodenum. Our detection of α-synuclein seeding activity in duodenum biopsies of Parkinson's disease patients suggests not only that such analyses may be useful in ante-mortem diagnosis, but also that the duodenum may be a source or a destination for pathological, self-propagating α-synuclein assemblies

Histological, immunohistochemical and anthropological study among mummies coming from different geographic areas

Following the discovery of two partially mummified corps in a Cathedral of Castelsardo, to better understand the mummification process it was decided to compare them with a peruvian mummy given to the University of Sassari in the late ‘800. The aim of this work is to compare tissue conservation status by a morphological study supported by anthropological analysis. Samples of skin and muscle were taken and rehydrated in Sadison’s solution to be subjected to histological and immunohistochemical procedures. To establish sex, death-age and height we worked following classical methods (Ferembach 1980; Uberlaker 1989; Meindl and Lovejoy 1985). Castelsardo’s mummies conditions are generally fair, although very variable in different points of the body: the former is male, with death-age between 45-55 years. His height is 171 cm. The latter female, with death-age between 60-67 years. Her height is 157 cm. She shows reduction of some intersomatic spaces, spondyloarthropathy and scoliosis. The muscular and cutaneous tissues show a good conservation, in particular the former presents a fibrillar structure well-preserved, boundaries free between cells. The peruvian mummy is male, with death-age older than 25 years. His height is 160 cm. He shows osteophytosis at lumbar area with a collapse of the body at 4th lumbar vertebra; his conditions are overall good however the tissues show a poor conservation where a structural organization cannot be distinguished. This study allowed us to obtain paleo and microanatomy informations and to describe the morphological characteristics of mummified tissues

Glucorticoid receptor in human cutaneous melanoma: immunohistochemical and immunofluorescence study

GR is a nuclear receptor which, when activated by its specific ligand, can act as a transcription factor that binds to glucocorticoid response elements (GRE) or negative GRE. It affects inflammatory responses, differentiation and cell proliferation. The ligand activated glucocorticoid receptor induces a G1 cell cycle arrest or apoptosis in immature thymocytes and impairs proliferation of fibroblasts of undifferentiated mammary epithelial cells. It impairs proliferation and differentiation of neural progenitor cells in vivo and in vitro. Glucocorticoids are widely used in cancer therapy and have cell type-specific pro- or antiapoptotic effects. In melanoma, however, the antitumor activity of glucocorticoids remains an open question. A recent report demonstrated that in mouse embryo tissue and in human undifferentiated cells, cytoplasmic accumulation of GR is determined by nestin in conjunction with vimentin, copolymerised into an intermediate filament system, and that this anchoring of GR to the nestin/vimentin etheromeric complex is related to the maintenance of a high proliferation rate. The aim of this study was to analyse the expression of subcellular GR in cutaneous melanoma by immunofluorescence, immunohistochemistry and laser scanning confocal microscopy and to evaluate any effect in melanoma progression. The results will be discussed

Course and Lethality of SARS-CoV2 Epidemic in Nursing Homes after Vaccination in Florence, Italy

Evidence on the effectiveness of SARS-CoV-2 vaccines in nursing home (NHs) residents is limited. We examined the impact of the BNT162b2 mRNA SARS-CoV-2 vaccine on the course of the epidemic in NHs in the Florence Health District, Italy, before and after vaccination. Moreover, we assessed survival and hospitalization by vaccination status in SARS-CoV-2-positive cases occurring during the post-vaccination period. We calculated the weekly infection rates during the pre-vaccination (1 October–26 December 2020) and post-vaccination period (27 December 2020–31 March 2021). Cox analysis was used to analyze survival by vaccination status. The study involved 3730 residents (mean age 84, 69% female). Weekly infection rates fluctuated during the pre-vaccination period (1.8%–6.5%) and dropped to zero during the post-vaccination period. Nine unvaccinated (UN), 56 partially vaccinated (PV) and 35 fully vaccinated (FV) residents tested SARS-CoV-2+ during the post-vaccination period. FV showed significantly lower hospitalization and mortality rates than PV and UV (hospitalization: FV 3%, PV 14%, UV 33%; mortality: FV 6%, PV 18%, UV 56%). The death risk was 84% and 96% lower in PV (HR 0.157, 95%CI 0.049–0.491) and FV (HR 0.037, 95%CI 0.006–0.223) versus UV. SARS-CoV-2 vaccination was followed by a marked decline in infection rates and was associated with lower morbidity and mortality among infected NH residents

Stormorken syndrome caused by a p.R304W STIM1 mutation: The first Italian patient and a review of the literature

Stormorken syndrome is a rare autosomal dominant disease that is characterized by a complex phenotype that includes tubular aggregate myopathy (TAM), bleeding diathesis, hyposplenism, mild hypocalcemia and additional features, such as miosis and a mild intellectual disability (dyslexia). Stormorken syndrome is caused by autosomal dominant mutations in the STIM1 gene, which encodes an endoplasmic reticulum Ca2+ sensor. Here, we describe the clinical and molecular aspects of a 21-year-old Italian female with Stormorken syndrome. The STIM1 gene sequence identified a c.910C T transition in a STIM1 allele (p.R304W). The p.R304W mutation is a common mutation that is responsible for Stormorken syndrome and is hypothesized to cause a gain of function action associated with a rise in Ca2+ levels. A review of published STIM1 mutations (n = 50) and reported Stormorken patients (n = 11) indicated a genotype-phenotype correlation with mutations in a coiled coil cytoplasmic domain associated with complete Stormorken syndrome, and other pathological variants outside this region were more often linked to an incomplete phenotype. Our study describes the first Italian patient with Stormorken syndrome, contributes to the genotype/phenotype correlation and highlights the possibility of directly investigating the p.R304W mutation in the presence of a typical phenotype

Case report: A novel ACTA1 variant in a patient with nemaline rods and increased glycogen deposition

BackgroundCongenital myopathies are a group of heterogeneous inherited disorders, mainly characterized by early-onset hypotonia and muscle weakness. The spectrum of clinical phenotype can be highly variable, going from very mild to severe presentations. The course also varies broadly resulting in a fatal outcome in the most severe cases but can either be benign or lead to an amelioration even in severe presentations. Muscle biopsy analysis is crucial for the identification of pathognomonic morphological features, such as core areas, nemaline bodies or rods, nuclear centralizations and congenital type 1 fibers disproportion. However, multiple abnormalities in the same muscle can be observed, making more complex the myopathological scenario.Case presentationHere, we describe an Italian newborn presenting with severe hypotonia, respiratory insufficiency, inability to suck and swallow, requiring mechanical ventilation and gastrostomy feeding. Muscle biopsy analyzed by light microscopy showed the presence of vacuoles filled with glycogen, suggesting a metabolic myopathy, but also fuchsinophilic inclusions. Ultrastructural studies confirmed the presence of normally structured glycogen, and the presence of minirods, directing the diagnostic hypothesis toward a nemaline myopathy. An expanded Next Generation Sequencing analysis targeting congenital myopathies genes revealed the presence of a novel heterozygous c.965 T &gt; A p. (Leu322Gln) variant in the ACTA1 gene, which encodes the skeletal muscle alpha-actin.ConclusionOur case expands the repertoire of molecular and pathological features observed in actinopathies. We highlight the value of ultrastructural examination to investigate the abnormalities detected at the histological level. We also emphasized the use of expanded gene panels in the molecular analysis of neuromuscular patients, especially for those ones presenting multiple bioptic alterations