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Clinical Laboratory Sep 2023Thiamine responsive megaloblastic anemia (TRMA) is a genetic disease caused by SLC19A2 gene mutation. This study aimed to preliminarily explore the relationship between...
BACKGROUND
Thiamine responsive megaloblastic anemia (TRMA) is a genetic disease caused by SLC19A2 gene mutation. This study aimed to preliminarily explore the relationship between endoplasmic reticulum stress (ERS)-PERK signaling pathway and the pathogenesis of hyperglycemia induced by TRMA.
METHODS
Islet β (INS.1 and β-TC-6) and HEK293T cell line models with stable overexpression of SLC19A2 and SLC19A2 (c.1409insT) were established. The cells were divided into empty virus group (control), wild-type group (overexpressed SLC19A2), and mutation group (overexpressed SLC19A2 (c.1409insT)). Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and western blotting were used to detect the expression levels of ERS-PERK signaling pathway-related proteins, including glucose-regulated protein 78 (GRP78), protein kinase R-like ER kinase (PERK), and eukaryotic initiation factor 2 (eIF2α), activating transcription factor 4 (ATF4), and C/EBP homologous protein (CHOP) in islet β cells. Protein localization was assessed by immunofluorescence staining.
RESULTS
Compared with the control group, the mRNA expression levels of SLC19A2 in wild-type and mutant islet β cells (INS.1 and β-TC-6) and HEK293T cells were significantly upregulated (all p < 0.05). Compared with the control group and the wild-type group, the mRNA expression levels of GRP78, PERK, eIF2α, ATF4, and CHOP were increased (all p < 0.05) in the mutant islet β cells; the protein expression levels of PERK, GRP78, and eIF2α were elevated (all p < 0.05). In addition, the results of immunofluorescence staining showed that SLC19A2 (c.1409insT) mutation changed the localization of the proteins in the cells. Thus, they were not located on the cell surface, but in the cytoplasm and nuclei, and protein aggregation occurred in the cytoplasm.
CONCLUSIONS
1. Islet β and HEK293Tcell lines, stably overexpressing SLC19A2 and SLC19A2 (c.1409insT) mutations, were successfully constructed. 2. SLC19A2 (c.1409insT) mutation could raise the expression levels of ERS-PERK signaling pathway-related proteins (GRP78, PERK, eIF2α, ATF4, and CHOP), and activate apoptosis pathway. 3. SLC19A2 (c.1409insT) mutation could change the localization of proteins and produce protein aggregation in cells. It could lead to protein misfolding and ERS, which would participate in the pathological mechanism of hyperglycemia induced by TRMA.
Topics: Humans; Endoplasmic Reticulum Chaperone BiP; HEK293 Cells; Protein Aggregates; Hyperglycemia; Anemia, Pernicious; Endoplasmic Reticulum Stress; Thiamine; RNA, Messenger; Membrane Transport Proteins
PubMed: 37702666
DOI: 10.7754/Clin.Lab.2023.230324 -
Internal Medicine Journal Apr 2024There are limited data relating to the effects of metformin-associated vitamin B12 deficiency on the risk of distal symmetrical polyneuropathy (DSPN) and megaloblastic...
BACKGROUND
There are limited data relating to the effects of metformin-associated vitamin B12 deficiency on the risk of distal symmetrical polyneuropathy (DSPN) and megaloblastic anaemia in well-characterised community-based cohorts.
AIMS
To assess inter-relationships between metformin therapy, vitamin B12 deficiency assessed using serum active B12 concentrations, and DSPN and anaemia in 1492 Fremantle Diabetes Study Phase 2 (FDS2) participants with type 2 diabetes.
METHODS
Prevalence rates of vitamin B12 deficiency (total <80 pmol/L, active <23 pmol/L) and borderline deficiency (total ≥80 and ≤200 pmol/L, active ≥23 and ≤35 pmol/L) were determined using baseline sera. The relationship between vitamin B12 status and both DSPN and anaemia was assessed using multivariable analyses.
RESULTS
Most FDS2 participants (94.4%) were vitamin B12 replete (total serum concentration >200 pmol/L, active >35 pmol/L), 2.0% were deficient (total <80 pmol/L, active <23 pmol/L) and the remainder (3.6%) borderline. Although metformin treatment increased the odds of deficiency (4.2%, 3.1% borderline) in a dose-dependent fashion (odds ratio (95% confidence interval) 39.4 (4.90-316) for >2000 mg daily compared with no treatment; P < 0.001), there was no significant association between vitamin B12 status and DSPN, anaemia (haemoglobin ≤130 g/L males, ≤120 g/L females), haemoglobin concentration or mean corpuscular volume (P ≥ 0.147). Metformin increased the likelihood of anaemia, especially at high doses, independent of vitamin B12 deficiency.
CONCLUSIONS
Since nutritional sources likely attenuate metformin-associated vitamin B12 malabsorption and its clinical sequelae in developed countries such as Australia, there is no need for routine/opportunistic serum vitamin B12 screening in metformin-treated patients.
Topics: Male; Female; Humans; Diabetes Mellitus, Type 2; Hypoglycemic Agents; Metformin; Vitamin B 12; Vitamin B 12 Deficiency; Anemia; Hemoglobins; Polyneuropathies
PubMed: 37665691
DOI: 10.1111/imj.16225 -
Gastroenterology Dec 2023
Topics: Humans; Stomach; Anemia; Upper Gastrointestinal Tract; Abdomen; Anemia, Pernicious; Stomach Neoplasms
PubMed: 37640254
DOI: 10.1053/j.gastro.2023.08.032 -
Cureus Jul 2023Myelodysplastic neoplasia (MDS) is a group of stem cell disorders involving ineffective hematopoiesis. It can be associated with an increased risk of progression toward...
Myelodysplastic neoplasia (MDS) is a group of stem cell disorders involving ineffective hematopoiesis. It can be associated with an increased risk of progression toward acute myeloid leukemia (AML). In Bahrain, MDS is the fifth most common primary hematologic malignancy. MDS has an annual incidence of up to 4 million cases. Some of the presenting signs and symptoms of MDS are often nonspecific, such as fatigue, pallor, malaise, fevers, bleeding, bruising, weight loss, and anorexia. Approximately 40% of patients with MDS progress to AML. This paper outlines a case of a 3-year-old Bahraini male (known to have sickle cell trait) who presented to the emergency department of Salmaniya Medical Complex with a five-day history of fever, congested throat, left ear pain, and abdominal pain. He had one episode of vomiting gastric content the previous day. He had previously gone to a private clinic with similar symptoms. Physical examination revealed a short neck and short stature, which was found to be below the 5 percentile. He had generalized pallor and hepatosplenomegaly. A blood smear showed leukopenia and normochromic normocytic anemia. There were excessive blasts found which consisted of 17% of nucleated cells and few granulopoietic cells. Erythropoiesis was active with a few showing mild megaloblastic changes. There were rare megakaryocytes noted. Moreover, the bone marrow aspirate showed two populations on dim CD45. The first population consisted of 3.15% on dim CD45 comprising of hematogones which brightly expressed CD19, HLA-DR, CD79a, and dim CD10. The second population consisted of 14.85% on dim CD45 which expressed CD34, CD13, CD117, HLA-DR, and dim CD7. Based on the peripheral blood smear and bone marrow immunophenotyping findings, a diagnosis of myelodysplastic syndrome with excessive blasts was made, which soon transformed into a diagnosis of AML. Furthermore, increased levels of dysplastic changes and percentage of blasts in the peripheral blood smear and bone marrow lead to a higher possibility of transformation into AML. As per the WHO classification, a diagnosis of MDS needs evaluation of the morphology of blood and bone marrow.
PubMed: 37593262
DOI: 10.7759/cureus.41988 -
Clinical Medicine Insights. Case Reports 2023Bardet-Biedl syndrome (BBS) also known as Laurence-Moon-Bardet-Biedl syndrome one of the rarely reported genetic disorder characterized by an intellectual disability,...
BACKGROUND
Bardet-Biedl syndrome (BBS) also known as Laurence-Moon-Bardet-Biedl syndrome one of the rarely reported genetic disorder characterized by an intellectual disability, limb, kidney abnormalities, obesity, and Rod-cone dystrophy. Other associated condition includes diabetes mellitus, hypertension, hypogonadism, facial dysmorphism, and congenital heart defects. This case highlights megaloblastic anemia associated with BBS.
CASE PRESENTATION
A 16-year-old female patient who had a moon face, truncal obesity, polydactyly, low IQ, and visual impairment presented with the complaint of shortness of breath and easy fatiguability. She had bilateral retinal pigmentosa in her eyes and her laboratory evaluation and bone marrow biopsy revealed megaloblastic anemia secondary to vitamin B12 deficiency. She received injectable vitamin B12, folate, and red cell contrate transfusion. Her symptoms improved and she was discharged with oral medication.
CONCLUSION
Megaloblastic anemia in BBS is rarely reported, further research is needed to find the exact cause that is necessary for proper management and better outcome.
PubMed: 37588947
DOI: 10.1177/11795476231193896 -
Analytical Biochemistry Oct 2023Inadequate folic acid intake is linked to diseases such as megaloblastic anemia, neural tube defects, and hyperhomocysteinemia, increasing the risk of vascular disease...
Inadequate folic acid intake is linked to diseases such as megaloblastic anemia, neural tube defects, and hyperhomocysteinemia, increasing the risk of vascular disease and thrombosis. Folic acid, a cofactor in various enzymes, can be produced by plants and bacteria, but not by humans and other animals. L-5-methyl-tetrahydrofolate (L-5-methyl-THF) is the primary dietary folate form, transported in circulation for cellular metabolism. Traditional methods of determining folic acid levels are unreliable and time-consuming. SenFol (Sensor for folic acid) is a fluorescence resonance energy transfer (FRET)-based nanosensor that we have developed by inserting folic acid-binding protein (FolT) as the folate detecting domain between the pair of enhanced cyan fluorescent protein (ECFP) and Venus. The developed sensor is highly specific, produces a quick signal, which is pH stable, and delivers precise, ratiometric readings in cell-based experiments. The projected affinity score of folic acid with FolT was -7.4 kcal/mol. The apparent affinity (K) of SenFol for folic acid is 28.49 × 10 M, with a detection range of 5 × 10 M to 5 × 10 M, and a maximum FRET ratio change of 0.45. WT SenFol, a highly efficient folic acid nanosensor, can dynamically detect intracellular folic acid content in E. coli, yeast, and HEK-293 T cells, confirming its potential.
Topics: Humans; Animals; Escherichia coli; Folic Acid; Fluorescence Resonance Energy Transfer; HEK293 Cells; Diagnostic Imaging; Saccharomyces cerevisiae
PubMed: 37586674
DOI: 10.1016/j.ab.2023.115285 -
Clinical Case Reports Aug 2023It is important for pregnant and breastfeeding women who adhere to a strict vegetarian diet to take appropriate steps to avoid vitamin B12 deficiency in their infants.
KEY CLINICAL MESSAGE
It is important for pregnant and breastfeeding women who adhere to a strict vegetarian diet to take appropriate steps to avoid vitamin B12 deficiency in their infants.
ABSTRACT
Vitamin B12 deficiency is rare during infancy. The initial symptoms of this deficiency are subtle and may include irritability, failure to thrive with a decline in growth rate, apathy, anorexia, refusal of solid foods, megaloblastic anemia, and developmental regression. The case presented here involves an 8-month-old male infant who showed neurological symptoms such as decreased activity, increased drowsiness, and reduced interaction with parents, which were ultimately linked to a deficiency of cobalamin (vitamin B12). Early recognition of this condition is critical because it is reversible. Therefore, pregnant and lactating women who follow a strict vegetarian diet should take necessary measures to prevent vitamin B12 deficiency in infants.
PubMed: 37554579
DOI: 10.1002/ccr3.7770 -
Cureus Jul 2023Introduction Thrombocytopenia is a commonly observed condition in clinical practice, and its diagnosis is often challenging due to numerous aetiologies and variations in...
Introduction Thrombocytopenia is a commonly observed condition in clinical practice, and its diagnosis is often challenging due to numerous aetiologies and variations in clinical presentation. Early identification of thrombocytopenia and its causes can help prevent life-threatening haemorrhagic manifestations. Methodology A prospective observational study was conducted at a tertiary care hospital from February 2019 to January 2020. This evaluation aimed to determine the causes and prevalence of thrombocytopenia in a tertiary care setting. Patients aged 15 or older with a platelet count of fewer than 150,000/ µL were eligible for inclusion in this evaluation. Investigations for aetiology detection were recommended. Results During the one-year study period, a total of 100 patients, including 58 males and 42 females, with thrombocytopenia were selected for the study. The most common age group affected by thrombocytopenia in this study was between 46 and 55 years old. The most common clinical manifestations observed were generalised weakness (70%), haemorrhagic manifestations (60%), fever (50%), joint pain (37%), splenomegaly (35%), headache (30%), breathlessness (23%), lymphadenopathy (22%), hepatomegaly (24%), and abdominal pain (12%). The most prevalent causes of thrombocytopenia were megaloblastic anaemia (19 cases), dengue fever (15 cases), malaria (11 cases), enteric fever (nine cases), immune thrombocytopenia (ITP) (eight cases), and leukaemia (seven cases). Bleeding was reported as a symptom of thrombocytopenia in 60% of individuals in this study. Conclusion In the study, thrombocytopenia was more common in people aged 46-55 years, and males were more commonly affected than females. Megaloblastic anaemia and infectious disease were the most common causes of thrombocytopenia. Bleeding manifestations were found in 60% of patients with thrombocytopenia.
PubMed: 37551236
DOI: 10.7759/cureus.41511 -
Nature Communications Jul 2023Many hematological diseases are characterized by altered abundance and morphology of blood cells and their progenitors. Myelodysplastic syndromes (MDS), for example, are...
Many hematological diseases are characterized by altered abundance and morphology of blood cells and their progenitors. Myelodysplastic syndromes (MDS), for example, are a group of blood cancers characterised by cytopenias, dysplasia of hematopoietic cells and blast expansion. Examination of peripheral blood slides (PBS) in MDS often reveals changes such as abnormal granulocyte lobulation or granularity and altered red blood cell (RBC) morphology; however, some of these features are shared with conditions such as haematinic deficiency anemias. Definitive diagnosis of MDS requires expert cytomorphology analysis of bone marrow smears and complementary information such as blood counts, karyotype and molecular genetics testing. Here, we present Haemorasis, a computational method that detects and characterizes white blood cells (WBC) and RBC in PBS. Applied to over 300 individuals with different conditions (SF3B1-mutant and SF3B1-wildtype MDS, megaloblastic anemia, and iron deficiency anemia), Haemorasis detected over half a million WBC and millions of RBC and characterized their morphology. These large sets of cell morphologies can be used in diagnosis and disease subtyping, while identifying novel associations between computational morphotypes and disease. We find that hypolobulated neutrophils and large RBC are characteristic of SF3B1-mutant MDS. Additionally, while prevalent in both iron deficiency and megaloblastic anemia, hyperlobulated neutrophils are larger in the latter. By integrating cytomorphological features using machine learning, Haemorasis was able to distinguish SF3B1-mutant MDS from other MDS using cytomorphology and blood counts alone, with high predictive performance. We validate our findings externally, showing that they generalize to other centers and scanners. Collectively, our work reveals the potential for the large-scale incorporation of automated cytomorphology into routine diagnostic workflows.
Topics: Humans; Myelodysplastic Syndromes; Anemia; Anemia, Megaloblastic; Blood Cells; Neutrophils
PubMed: 37474506
DOI: 10.1038/s41467-023-39676-y