Bone Health Impairment in Patients with Hemoglobinopathies: From Biological Bases to New Possible Therapeutic Strategies.

Hemoglobinopathies are monogenic disorders affecting hemoglobin synthesis. Thalassemia and sickle cell disease (SCD) are considered the two major hemoglobinopathies. Thalassemia is a genetic disorder and one of the major hemoglobinopathies determined by an impairment of globin chain production, which causes an alteration of erythropoiesis, an improvement in hemolysis, and an alteration of iron homoeostasis. In SCD, the mutations are on the ß-globin chain of hemoglobin which results in a substitution of glutamic acid by valine with consequent formation of Hemoglobin S (HbS). Several factors are involved in bone metabolism alteration in patients with hemoglobinopathies, among them hormonal deficiency, bone marrow hyperplasia, iron overload, inflammation, and increased bone turnover. Bone metabolism is the result of balance maintenance between bone deposition and bone resorption, by osteoblasts (OBs) and osteoclasts (OCs). An impairment of this balance is responsible for the onset of bone diseases, such as osteoporosis (OP). Therefore, here we will discuss the alteration of bone metabolism in patients with hemoglobinopathies and the possible therapeutic strategies to contain and/or counteract bone health impairment in these patients, taking into consideration not only the pharmacological treatments already used in the clinical armamentarium, but also the new possible therapeutic strategies.

Role of Nutraceuticals in Counteracting Inflammation in In Vitro Macrophages Obtained from Childhood Cancer Survivors.

The advancement of anti-cancer therapies has markedly improved the survival rate of children with cancer, making them long-term childhood cancer survivors (CCS). Nevertheless, these treatments cause a low-grade inflammatory state, determining inflamm-aging and, thus, favoring the early onset of chronic diseases normally associated with old age. Identification of novel and safer therapeutic strategies is needed to counteract and prevent inflamm-aging. Macrophages are cells involved in immune and inflammatory responses, with a pivotal role in iron metabolism, which is related to inflammation. We obtained macrophages from CCS patients and evaluated their phenotype markers, inflammatory states, and iron metabolism by Western blotting, ELISA, and iron assays. We observed a strong increase in classically activated phenotype markers (M1) and iron metabolism alteration in CCS, with an increase in intracellular iron concentration and inflammatory markers. These results suggest that the prevalence of M1 macrophages and alteration of iron metabolism could be involved in the worsening of inflammation in CCS. Therefore, we propose macrophages and iron metabolism as novel therapeutic targets to counteract inflamm-aging. To avoid toxic regimens, we tested some nutraceuticals (resveratrol, curcumin, and oil-enriched lycopene), which are already known to exert anti-inflammatory properties. After their administration, we observed a macrophage switch towards the anti-inflammatory phenotype M2, as well as reductions in pro-inflammatory cytokines and the intracellular iron concentration. Therefore, we suggest-for the first time-that nutraceuticals reduce inflammation in CCS macrophages through a novel anti-inflammatory mechanism of action, modulating iron metabolism.

Chondroblastoma of proximal tibia diagnosed by arthroscopy-guided biopsy: a case report.

Chondroblastoma considered a rare form of osseous neoplasm contributes less than 1% of all bone tumours. It is typically found in young patients with chief complaints of moderate pain with joint stiffness. It develops as a lytic lesion in the epiphysis of long bones which might spread to the metaphysis. We report a case of an 18-year-old patient who presented with progressive right knee pain which aggravated with movements. Investigations included X-ray, magnetic resonance imaging (MRI), and biochemical assessment. A focal, well-defined, lesion in the upper end of the tibia with surrounding marrow oedema was observed and diagnostic arthroscopy was taken for management. Histopathology of specimen observed chondroblasts proliferation with areas of mature cartilage, and giant cells confirming intrasynovial chondroblastoma. Usually, surgery is the treatment of choice; however, possibilities of the secondary bone cyst, haemosiderin deposition on the joint, etc., make treatment approaches uncertain. Diagnostic arthroscopy is a rare but essential modality in such cases due to better visuals, complete tumour excision, and combination with adjuvant therapies. Chondroblastoma, if untreated, proves detrimental, hence, a thorough evaluation is critical for overall better outcomes.

Metastatic mastery: the uncommon journey of papillary thyroid carcinoma to the brain (a case report).

Papillary thyroid carcinoma is one of the most common thyroid malignancy, often has excellent prognosis and low incidence of distant metastatic conditions. Brain metastases from papillary thyroid carcinoma has a rare occurrence, with patients presenting with non-specific symptoms such as headaches, cognitive changes etc., and poor survival outcomes. The standard protocol for diagnosis and treatment remains controversial. We report a patient who presented with cerebral metastasis prior to the diagnosis of papillary thyroid carcinoma, review the current literature, and explain our approach on the basis of clinical, pathological, and radiological data. A 60-year-old hypertensive male presented with lower back pain, bilateral lower limb weakness, occasional episodes of frontal headache and personality changes. The diagnostic evaluation included computed tomography (CT) scan, magnetic resonance imaging (MRI) with and without contrast enhancement, and color Doppler. Intra-axial complex solid cystic mass lesion in the right parieto-occipital region with significant perilesional oedema, and imaging characteristic of neoplastic etiology were observed. He was posted for excision of tumor and underwent right occipital craniotomy. Histopathological analysis of the surgical specimen confirmed papillary carcinoma of the thyroid gland. Brain metastases from thyroid malignancy is a sign of detrimental prognosis, hence, thorough clinical, radiological and pathological evaluation for rapid detection is critical. Neurosurgical removal along with radiotherapy should be considered as treatment of choice. The information obtained contributes towards better management and overall long-term outcomes.

ACE2 protein expression in lung tissues of severe COVID-19 infection.

Angiotensin-converting enzyme 2 (ACE2) is a key host protein by which severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) enters and multiplies within cells. The level of ACE2 expression in the lung is hypothesised to correlate with an increased risk of severe infection and complications in COrona VIrus Disease 2019 (COVID-19). To test this hypothesis, we compared the protein expression status of ACE2 by immunohistochemistry (IHC) in post-mortem lung samples of patients who died of severe COVID-19 and lung samples obtained from non-COVID-19 patients for other indications. IHC for CD61 and CD163 was performed for the assessment of platelet-rich microthrombi and macrophages, respectively. IHC for SARS-CoV-2 viral antigen was also performed. In a total of 55, 44 COVID-19 post-mortem lung samples were tested for ACE2, 36 for CD163, and 26 for CD61, compared to 15 non-covid 19 control lung sections. Quantification of immunostaining, random sampling, and correlation analysis were used to substantiate the morphologic findings. Our results show that ACE2 protein expression was significantly higher in COVID-19 post-mortem lung tissues than in controls, regardless of sample size. Histomorphology in COVID-19 lungs showed diffuse alveolar damage (DAD), acute bronchopneumonia, and acute lung injury with SARS-CoV-2 viral protein detected in a subset of cases. ACE2 expression levels were positively correlated with increased expression levels of CD61 and CD163. In conclusion, our results show significantly higher ACE2 protein expression in severe COVID-19 disease, correlating with increased macrophage infiltration and microthrombi, suggesting a pathobiological role in disease severity.

Multiplex fluorescence in situ hybridization testing for anaplastic lymphoma kinase and c-ros oncogene 1 gene rearrangements on cytology smears in lung adenocarcinomas: comparative study with formalin-fixed paraffin-embedded sections.

INTRODUCTION: Multiplex anaplastic lymphoma kinase (ALK)/c-ros oncogene 1 (ROS1) fluorescence in situ hybridization (FISH) probes conserve tissue by analyzing both ALK and ROS1 gene rearrangements (ALK-R/ROS1-R) in a single test. The positivity cutoffs have been validated on formalin-fixed, paraffin-embedded (FFPE) tissue sections and not tested on non-cell block (CB) cytology preparations. We sought to validate non-CB cytology preparations for the detection of ALK-R/ROS1-R using multiplex ALK/ROS1 FISH probes by comparing the results with matched FFPE results. MATERIALS AND METHODS: During the 3.5-year study period, FISH using the FlexISH ALK/ROS1 DistinguISH Probe (ZytoVision) was performed in non-CB cytology preparations of patients for whom FISH on FFPE sections was performed. RESULTS: A total of 20 patients had one or more non-CB cytology preparations (n = 27) suitable for FISH analysis. These comprised direct smears (n = 17), smears from centrifuged effusion pellets (n = 8), cytospin smears (n = 1), and biopsy imprint smears (n = 1). These had been fixed in 95% ethanol (n = 18) or air dried (n = 9), and stained with Papanicolaou (n = 14), May-Grünwald-Giemsa (n = 9), immunocytochemistry (n = 3), or hematoxylin and eosin (n = 1). The median archival time was 1 year. Successful FISH results were achieved in 14 samples (6 with ALK-R, 2 with ROS1-R, 6 negative) and were concordant with the FFPE FISH results for 13 of 14 cases. The single case with discordant results between cytology and FFPE FISH showed ALK-R on cytology concordant with positive ALK D5F3 companion diagnostics assay results and was considered a false-negative FFPE FISH result. FISH failure occurred mainly in the older archived slides because of overdigestion (n = 5), hybridization failure (n = 5), or excessive background fluorescence (n = 3). CONCLUSIONS: Non-CB cytology smears are highly suitable for multiplex FISH analysis with 100% concordance with FFPE FISH and/or ALK D5F3 companion diagnostics assay results.

Yoga improves mitochondrial health and reduces severity of autoimmune inflammatory arthritis: A randomized controlled trial.

BACKGROUND: Oxidative stress (OS) and mitochondrial alterations have been implicated in the pathogenesis of rheumatoid arthritis (RA). Various environmental triggers like air pollutants, smoking, unhealthy social habits and sedentary lifestyle induce OS, which may compromise mitochondrial integrity. This trial was designed to explore the effect of 8-weeks yoga practice on mitochondrial health and disease severity in an active RA group compared with a usual-care control group. METHODS: A total of 70 subjects were randomized into two groups: yoga group and non-yoga group. Mitochondrial health was assessed by calculation of mitochondrial DNA copy number (mtDNA-CN), OS markers, mitochondrial activity, mitochondrial membrane potential (ΔΨm), circadian rhythm markers and transcripts associated with mitochondrial integrity: AMPK, TIMP-1, KLOTHO, SIRT-1, and TFAM. Parameters of disease activity and disability quotient were also assessed by disease activity score - erythrocyte sedimentation rate (DAS28-ESR) and health assessment questionnaire-disability index (HAQ-DI), respectively. RESULTS: In yoga group, there was a significant upregulation of mtDNA-CN, mitochondrial activity markers, ΔΨm, and transcripts that maintain mitochondrial integrity after 8-weeks of yoga. There was optimization of OS markers, and circadian rhythm markers post 8-weeks practice of yoga. Yoga group participants showed significant improvements in DAS28-ESR (p < 0.05) and HAQ-DI (p < 0.05) over the non-yoga group. CONCLUSION: Adoption of yoga by RA patients holds the key to enhance mitochondrial health, improve circadian rhythm markers, OS marker regulation, upregulation of transcripts that maintain mitochondrial integrity, reduce disease activity and its associated consequences on health outcome and hence can be beneficial as an adjunct therapy.