Beta2-glycoprotein I Research Articles

Breaking the Pump: Targeting the Sodium-Potassium Pump As a Therapeutic Strategy in Acute Myeloid Leukemia

Acute myeloid leukemia (AML) is a heterogenous hematologic malignancy with poor overall survival despite intensive cytotoxic chemotherapy and newer targeted agents. With a 5-year survival rate of only 30% in adults with AML, new therapeutic strategies are needed. Tissue-specific differences in the expression of paralog genes, which are not essential in most cell types due to the buffering effect of the partner pair, could make for highly selective gene dependencies. To identify novel, selective paralogous targets for AML, we integrated the Cancer Dependency Map, which provides genome-scale CRISPR/Cas9 depletion screening data in hundreds of cancer cell lines, with numerous databases characterizing protein-protein interactions, paralog relationships, and gene expression in cancer models. Here, we identified ATP1B3 as a context-specific, paralog related dependency in AML. ATP1B3 is the smaller glycoprotein beta subunit of the sodium-potassium pump (Na/K-ATP pump). Together with the catalytic alpha subunit ATP1A1, it forms a heterodimer located in the plasma membrane, regulating the electrochemical gradient through the transport of Na and K ions across the membrane. The Na/K-ATP pump is of vital importance in the maintenance of cellular homeostasis and membrane potential. While ATP1A1 carries out the ion transport and enzymatic activity, making it a common essential gene (a gene which ranks among the most depleted genes in at least 90% of screened cell lines), ATP1B3 is not essential in most cancer cells. The beta subunit is important for intracellular transport and stabilization of the alpha subunit in the membrane, but it has three paralogs: ATP1B3 showing similar expression patterns across tissues, ATP1B2 exclusively expressed in brain and retina, and ATP1B1 showing differential expression across tissues. ATP1B1 has uniformly low expression in hematological malignancies and low ATP1B1 expression is the strongest predictor of ATP1B3 dependency (Figure 1). Using CRISPR/Cas9 knockout and competitive growth assays, we showed that knockout of ATP1B3 in ATP1B1 low-expressing AML cells indeed leads to synthetic lethality. Knockout of ATP1B3 significantly decreased number and size of colonies in a patient-derived xenograft (PDX) model of AML to a similar degree as the knockout of the common essential gene ATP1A1. Moreover, in these AML models, the absence of both paralogs of the beta subunit resulted in the destabilization of their binding partner ATP1A1. In contrast, expression of ATP1B1 stabilized ATP1A1 despite ATP1B3 knockout in solid tumors and in ATP1B1-overexpressing AML cells. In vivo, using an orthotopic AML cell line mouse model and an AML PDX model, ATP1B3 knockout resulted in lower leukemia burden in comparison to ATP1B1 knockout or a non-targeting control guide. To determine if ATP1B1 and ATP1B3 share primarily redundant functions, we utilized BioID (proximity-dependent biotin identification) to identify their interaction partners. We discovered that in AML, there is significant overlap of the ATP1B1 and ATP1B3 binding partners, and ATP1A1 is among the strongest. Enrichment of gene sets involved in protein folding in the endoplasmic reticulum are also consistent with the important role of the beta subunits in proper ATP1A1 folding and stabilization. Taken together, we identified ATP1B3 as a selective dependency in AML. We propose that when ATP1B1 is poorly expressed, elimination of ATP1B3 leads to the destabilization of the sodium-potassium pump including ATP1A1. ATP1B3 is a potential tumor-selective therapeutic target for AML and other hematologic malignancies with low expression of ATP1B1. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Apolipoprotein H: a novel regulator of fat accumulation in duck myoblasts

Apolipoprotein H (APOH) primarily engages in fat metabolism and inflammatory disease response. This study aimed to investigate the effects of APOH on fat synthesis in duck myoblasts (CS2s) by APOH overexpression and knockdown. CS2s overexpressing APOH showed enhanced triglyceride (TG) and cholesterol (CHOL) contents and elevated the mRNA and protein expression of AKT serine/threonine kinase 1 (AKT1), ELOVL fatty acid elongase 6 (ELOVL6), and acetyl-CoA carboxylase 1 (ACC1) while reducing the expression of protein kinase AMP-activated catalytic subunit alpha 1 (AMPK), peroxisome proliferator activated receptor gamma (PPARG), acyl-CoA synthetase long chain family member 1 (ACSL1), and lipoprotein lipase (LPL). The results showed that knockdown of APOH in CS2s reduced the content of TG and CHOL, reduced the expression of ACC1, ELOVL6, and AKT1, and increased the gene and protein expression of PPARG, LPL, ACSL1, and AMPK. Our results showed that APOH affected lipid deposition in myoblasts by inhibiting fatty acid beta-oxidation and promoting fatty acid biosynthesis by regulating the expression of the AKT/AMPK pathway. This study provides the necessary basic information for the role of APOH in fat accumulation in duck myoblasts for the first time and enables researchers to study the genes related to fat deposition in meat ducks in a new direction.

A RARE CASE OF ANTINEUTROPHIL CYTOPLASMIC ANTIBODY (ANCA)-NEGATIVE PAUCI- IMMUNE FOCAL NECROTIZING AND CRESCENTIC GLOMERULONEPHRITIS PRESENTING WITH DIFFUSE ALVEOLAR HEMORRHAGE

A RARE CASE OF ANTINEUTROPHIL CYTOPLASMIC ANTIBODY (ANCA)-NEGATIVE PAUCI- IMMUNE FOCAL NECROTIZING AND CRESCENTIC GLOMERULONEPHRITIS PRESENTING WITH DIFFUSE ALVEOLAR HEMORRHAGE

A CASE OF BORDETELLA BRONCHISEPTICA AND PNEUMOCYSTIS JIROVECI PNEUMONIA IN A YOUNG PATIENT

A CASE OF BORDETELLA BRONCHISEPTICA AND PNEUMOCYSTIS JIROVECI PNEUMONIA IN A YOUNG PATIENT

PRESENTATION OF ACUTE CEREBROVASCULAR ACCIDENT IN A YOUNG ADULT WITH THROMBOTIC THROMBOCYTOPENIA PURPURA (TTP)

PRESENTATION OF ACUTE CEREBROVASCULAR ACCIDENT IN A YOUNG ADULT WITH THROMBOTIC THROMBOCYTOPENIA PURPURA (TTP)

P17 All in the Marrow - A case of Primary Bone Marrow Oedema

Introduction/BackgroundPrimary bone marrow oedema syndrome is an elusive and less well-defined entity. Whether Rheumatologists should consider it as a stand alone diagnosis, is debatable. It possibly would be best described as an MRI feature which could be a finding in a number of diseases which would include the initial phases of Osteonecrosis of the bone, Rheumatoid Arthritis, Spondyloarthritis, Enthesitis related, Post traumatic, OA, Infections and Cancers. The treatment options become constricted due to the paucity of evidence.Rheumatologists need to consider this as an area of unmet need with development of consensus classification criteria and treatment approaches.Description/Method27-year-old male, Height 174 cms Weight 90 Kgs BMI 29 Kg/m2, became symptomatic in Jan 2022 with complains of pain in the both hip joints & groin regions, pain became excruciating and he became bed-bound, with early morning stiffness lasting approximately 45 mins.Had received steroids for COVID infection in August 2020.InvestigationsHb 13.5gm/dl TLC 7000/mm3 Platelet 400 × 103/mm3Sr Bil 0.8mg/dl AST 16 IU/L. ALT 24 IU/L Sr Creatininine 1.1mg/dlBlood Sugar Levels, Fasting 89 mg/dl Post Prandial 102 mg/dlESR 10 mm in 1st hour by Wintrobes method CRP Quantitative 29.38mg/LHLA B27 by PCR Negative, RF Negative, ACCP NegativeSerum, IgG, Beta2 Glycoprotein 1.44 SGU Serum, IgM, Beta2 Glycoprotein 3.44 SGUSerum, IgG, Cardiolipin antibody 8.4 GPL Serum, IgG, Cardiolipin antibody 17.45 GPLLupus anticoagulant by DRVVT NegativeSr Cholesterol 211mg/dl HDL 29 mg/dl LDL 156mg/dl TGs 130 mg/dlMRI Hips & SI joints Transient bone marrow oedema/osteopenia of bilateral hip.PET CT Increased metabolic activity in both hip jointsBone Scan (99mTcMDP) Increased vascularity in perfusion phase, increased accumulation in soft tissue in blood pool phase and increased uptake in bilateral Hip joints in skeletal phase scan, suggestive of CRPS Type-I.ManagementWas initially managed with Tab Etoricoxib 90mg BD, also started on Tab Sulphaslazine and Tab Methotrexate. However, when he had no symptomatic relief he was administered Inj Infliximab on 12 May 2022 and a second dose on 9 June 2022. He had excellent pain relief after the 1st dose, however after 10 days of the administration, he again began experiencing pain especially after walking. He also had pain in the knees on this occasion.He was also administered Inj Zoledronic 4mg on 23 May 2022.He is at present not requiring any NSAIDs over the last 1 month.Discussion/ResultsThe patient having presented with excruciating and debilitating pain was worked up and evaluation revealed features of bone marrow oedema on MRI which was corroborated with bone scan and PET CT imaging. The acute phase reactant CRP was also significantly elevated. The patient also gave history of early morning stiffness lasting approximately 45 mins. Hence an underlying Inflammatory process such as Spondyloarthritis(Peripheral) with enthesitis was considered. The confounding factors were the pain which worsened on mobilization, HLA B27 negative status, Rheumatoid Factor and ACCP negative status and past history of having received IV Corticosteroids for COVID infection in August 2020.In view of the debilitating pain and a working diagnosis of Spondyloarthritis, he was started on NSAIDs along with rest, initially, followed by conventional synthetic disease modifying agents in Rheumatic disease followed by biologic synthetic disease modifying agent – Inj Infliximab.The thought process was to avoid prolonged NSAID use to prevent the associated side effects.However, since Avascular Necrosis of the Femoral head is a very likely possibility, the patient is planned to be kept under close follow up.Key learning points/ConclusionCollaborative efforts between the Departments of Nuclear Medicine, Radiology, Orthopaedics and Rheumatology are crucial in the early detection and approach to cases of Bone Marrow oedema.Avascular necrosis of head of Femur is a far more common entity and must be kept in mind even when a diagnosis of Bone Marrow oedema syndrome is being entertained.Diagnosis of Bone Marrow oedema syndrome must be entertained only as a diagnosis of exclusion.Continued follow up and regular imaging must be pursued rigorously in patients diagnosed with Bone Marrow oedema syndromes.There is a requirement to document acute phase reactants such as CRP and ESR in patients diagnosed with Avascular necrosis of bone as this data could help us differentiate AVN from Primary Bone marrow oedema in the early stages.

Alcohol-dependent downregulation of apolipoprotein H exacerbates fatty liver and gut microbiota dysbiosis in mice

BackgroundAlcohol-related liver disease (ALD) is a major chronic liver ailment caused by alcohol overconsumption and abuse. Apolipoprotein H (APOH) participates in lipid metabolism and might have a potential regulatory role in ALD. Therefore, this study aimed to explore the effects of ApoH on alcohol-induced liver injury and gut microbiota dysbiosis.MethodsApoH−/− mice were generated and the synergic alcoholic steatohepatitis mouse model was constructed, which were used to assess liver function and pathological changes.ResultsApoH−/− mice clearly exhibited spontaneous steatohepatitis. Severe hepatic steatosis was observed in alcohol-fed WT and ApoH−/− mice, in which ApoH expression was reduced post alcohol consumption. Moreover, RNA-seq and KEGG pathway analyses indicated that differential expression genes enriched in lipid metabolism and oxidation–reduction process between in alcohol-fed ApoH−/− mice and pair-fed control mice. Finally, gut microbiota diversity and composition were assessed by 16S rRNA Illumina next-generation sequencing. Alpha diversity of enterobacteria was lower in ApoH−/− mice with ethanol feeding than in ethanol-fed WT mice and all control-fed mice (P < 0.05). Moreover, KEGG enrichment analysis, using PICRUSt software, revealed that metabolic functions were activated in the gut microorganisms of ApoH−/− mice with ethanol feeding (P < 0.05).ConclusionsAlcohol-downregulated ApoH expression, leading to the progress of fatty liver disease and gut microbiota dysbiosis.

IgA Antiphospholipid Antibodies in Antiphospholipid Syndrome and Systemic Lupus Erythematosus.

Objective: To define the role of IgA antibodies to cardiolipin (aCL) and IgA antibodies to beta-2 glycoprotein 1 (anti-β2-GP1) in the development of vascular complications in patients with antiphospholipid syndrome (APS) and systemic lupus erythematosus (SLE). Material and methods: A total of 187 patients with one of the following diagnoses: primary APS (PAPS), probable APS, SLE with APS, and SLE without APS. The comparison group consisted of 49 patients with other rheumatic diseases (RD), the control group included 100 relatively healthy individuals (without RD, oncological pathology, and infectious diseases). All patients underwent standard clinical, laboratory, and instrumental examinations before being included in the study and during follow-up. The aPL study included the determination of IgG/IgM aCL, IgG/IgM anti-β2-GP1 by enzyme-linked immunosorbent assay (ELISA), IgG/IgM/IgA aCL, IgG/IgM/IgA anti-β2-GP1 by chemiluminescence analysis (CLA), and lupus anticoagulant (LA). Results: IgA aCL were detected in 75 (40%) of the 187 patients with APS and SLE, in none of the comparison group, and in 2 (2%) of the control one. IgA anti-β2-GP1 were detected in 63 (34%) of the 187 patients with APS and SLE, in none of the patients in the comparison group, and in one (1%) of the control group. The prevalence of IgA aCL and IgA anti-β2-GP1 and their levels were statistically significantly higher in patients with APS (PAPS and SLE + APS) than the levels in patients with SLE and those of the comparison and control groups (p < 0.05). IgA aCL and IgA anti-β2-GP1 were significantly associated with thrombosis in APS (χ2 = 4.96; p = 0.02 and χ2 = 4.37; p = 0.04, respectively). The risk of thrombosis was 2.04 times higher in patients with positive IgA aCL than in patients without these antibodies, as well as in patients with positive IgA anti-β2-GP1; it was twice as high as in patients without antibodies. There was a high specificity of IgA aCL and IgA anti-β2-GP1 for both the diagnosis of APS and its clinical manifestations, despite a low sensitivity. Conclusions: The study revealed a relationship of thrombosis and APS with IgA aCL and IgA anti-β2-GP1. There was a high specificity of IgA aCL and IgA anti-β2-GP1 (95% and 93%, respectively) for the diagnosis of APS with a low sensitivity (54% and 44%, respectively). There were no patients with isolated positivity of IgA aCL and IgA anti-β2-GP1.

Non-criteria antiphospholipid antibodies and pediatric rheumatic disease: a case series

BackgroundNon-criteria antiphospholipid antibodies (NC-aPL) are a relatively undefined subgroup of antiphospholipid antibodies (aPL). Knowledge about NC-aPL in adults is limited and even less is known in pediatric patients. Routine tests for antiphospholipid syndrome (APS)—a clinical state marked by the presence of aPL in association with vascular thrombosis—usually include lupus anticoagulant (LAC), anti-cardiolipin (aCL) and -beta-2 glycoprotein I (aβ2GPI). LAC is a functional screen for prothrombotic aPL, while the latter tests identify specific autoantibodies. Specific targets of NC-aPL include, but are not limited to, phosphatidylethanolamine, phosphatidylserine, and prothrombin.Presentation of casesWe present single-center data from eight pediatric patients with NC-aPL identified during a three-year period. All patients had presenting features raising suspicion for APS. Most patients were female with a primary rheumatic disease. One patient had a stroke. Another patient had alveolar hemorrhage and pulmonary hypertension. Raynaud’s phenomenon, rashes involving distal extremities, and headaches were common. Most patients had a positive LAC, yet their routine aPL tests were negative, prompting testing for NC-aPL.ConclusionsOur findings suggest NC-aPL are associated with typical signs and symptoms of APS in pediatric patients. Pediatricians and pediatric subspecialists should consider NC-aPL when clinical suspicion is high and routine aPL tests are negative, particularly when LAC is positive. While guidelines for NC-aPL do not yet exist for children or adults, these autoantibodies have pathogenic potential. Actionable items could include evaluation for the presence of other (primary) rheumatic diseases, and consultation with hematologists and/or obstetricians regarding anticoagulation/platelet inhibition and thrombosis education. Future guidelines regarding NC-aPL will only be generated by gathering more data, ideally prospectively.

Pathogenetic significance of antiphospholipid antibodies in the development of placenta-associated complications in pregnant women with thrombocytopenia

BACKGROUND: Thrombocytopenia is one of the most common cytopenic syndromes that complicate pregnancy. It is considered a typical hematological criterion and the second most common manifestation of antiphospholipid syndrome. In recent years, the relationship between the development of thrombotic and placenta-associated pregnancy complications and the circulation of antiphospholipid antibodies has been actively studied, which makes it necessary to search for new methods for hemostasis correction and for prevention of the development of obstetric complications.&#x0D; AIM: The aim of this study was to assess the incidence of antiphospholipid antibodies in pregnant women with thrombocytopenia in order to determine the risk of obstetric complications and adverse pregnancy outcomes depending on the presence of antiphospholipid antibodies in the examined women.&#x0D; MATERIALS AND METHODS: 299 pregnant women were included in this multicenter prospective study: the main study group (n = 249) consisted of patients with thrombocytopenia, and the control group (n = 50) comprised women with normal platelet counts during a physiological pregnancy. All participants underwent a complete clinical and anamnestic examination and a study of the hemostasis system. To determine antibodies to cardiolipin, beta-2 glycoprotein 1, annexin V, and prothrombin (IgG/IgM/IgA), we performed enzyme-linked immunosorbent assay in 84 pregnant women with thrombocytopenia and in 25 women in the control group. Lupus anticoagulant was determined in 123 pregnant women of the main group and 25 women in the control group using an ACL-200 coagulometer (Instrumentation Laboratory, Spain).&#x0D; RESULTS: A comparative analysis of antiphospholipid antibodies detection has shown that in pregnant women with thrombocytopenia, lupus anticoagulant was detected in every third case (35.8% of cases, 44/123 women), which is more common than in the control group (8.0% of cases, 2/25 women; p = 0.008). Antibodies to beta-2 glycoprotein 1 were detected in pregnant women with thrombocytopenia in every fourth case (27.4% of cases, 23/84 women), and in the control group only in 4% of cases (1/25 women; p = 0.013). Antibodies to cardiolipin were also more often detected in pregnant women with thrombocytopenia (23.8% of cases, 20/84 women) compared to the control group (4% of cases, 1/25 women; p = 0.040). Based on the data obtained with V Cramer, we established medium-strength direct correlations between criterial antiphospholipid antibodies circulation and the development of placenta-associated complications of pregnancy (p 0.05; 0.2 V 0.4). We also identified statistically significant risks of developing placenta-associated complications in pregnant women with circulating antiphospholipid antibodies (95 % confidence interval excludes 1).&#x0D; CONCLUSIONS: Thrombocytopenia during pregnancy develops more often in women with antiphospholipid antibodies. Antiphospholipid antibodies circulation elevates coagulation potential and activates intravascular coagulation, resulting in an increased risk of placenta-associated complications of pregnancy, which requires hemostasis correction.

Antiphospholipid antibodies and vitamin D deficiency in COVID-19 infection with and without venous or arterial thrombosis: A pilot case-control study.

Coronavirus disease-2019 (COVID-19) is associated with thromboembolism. Antiphospholipid antibody (APLa) formation is one of the mechanisms. Vitamin D deficiency has been associated with thrombosis in antiphospholipid antibody syndrome. Measure APLa and vitamin D in hospitalized COVID-19 patients with and without thrombosis to evaluate if thromboembolism is associated with concomitant APLa and vitamin D deficiency. Case-control study. Hospitalized COVID-19 patients with a thromboembolic event (ischemic stroke, myocardial infarction, deep venous thrombosis/pulmonary embolism, Cases n = 20). Controls (n = 20): Age, sex-matched without thromboembolic events. Patients with autoimmune disorders, antiphospholipid antibody syndrome, thrombophilia, anticoagulation therapy, prior thromboembolism, chronic kidney disease 3b, 4, end-stage renal disease, and malignancy were excluded. Given the limited current literature on the role of concomitant antiphospholipid antibodies and vitamin D deficiency in causing venous and/or arterial thrombosis in hospitalized COVID-19 patients, we enrolled 20 patients in each arm. Anti-cardiolipin IgG/IgM, beta-2 glycoprotein-1 IgG/IgM, lupus anticoagulant and vitamin D levels were measured in both groups. Cases were 5.7 times more likely to be vitamin D deficient (OR:5.7, 95% CI:1.3-25.6) and 7.4 times more likely to have any one APLa (OR:7.4, 95% CI: 1.6-49.5) while accounting for the effects of sex. Patients with both APLa and vitamin D deficiency had significantly more thrombosis compared to patients who were antibody positive without vitamin D deficiency (100% vs 47.4%; p = 0.01). Thrombosis in COVID-19 was associated with concomitant APLa and vitamin D deficiency. Future studies in COVID-19 should assess the role of vitamin D in reducing thrombosis.

Neurological and Neuropsychiatric Manifestations of Antiphospholipid-Antibody Syndrome (APS).

Antiphospholipid antibody syndrome (APS) is an autoimmune disorder mediated by the presence of a group of autoantibodies, specifically the anticardiolipin antibody (aCL), the beta-2 glycoprotein I (β2GPI), and the lupus anticoagulant (LA). Patients diagnosed with antiphospholipid antibody syndrome (APS) present with many symptoms, the most common being the consequence of thrombotic events that can be catastrophic and lead to mild to severe residual disabilities over a significant amount of time and can impair the quality of life. These events are often present in the younger population. Many times, these thrombotic events are heralded by a spectrum of psychiatric symptoms, which when worked up in the right direction may hint toward an oncoming thrombotic event and may potentially prevent those events by prompting primary prophylaxis treatment by the treating physician. In this review, we aim to comprehensively put forth the many neurological and neuropsychiatric manifestations of APS, their pathology, and management.

Investigation of proteomic profiles in canine lymphoma using tandem mass tag-based quantitative proteomics approach.

Background and Aim:Specific tumor biomarkers are useful for the early diagnosis of cancer or can predict the recurrence of neoplastic disease in humans and animals. Lymphoma in dogs could be classified into B-, T-, and NK-cell origins. T-cell lymphoma has the worst prognosis with a shorter survival time and disease-free interval. This study aimed to identify the differential serum protein expressions of canine B- and T-cell lymphomas compared with healthy dogs using a tandem mass tag (TMT)-based quantitative proteomics.Materials and Methods:Serum samples were collected from 20 untreated canine lymphomas (14 B-cells and 6 T-cells) and four healthy control dogs. Sera peptides from each sample were processed for TMT 10-plex tagging and analyzed using liquid chromatography-mass spectrometry (MS). Differential proteome profiling was then compared between lymphoma and control.Results:We discovered 20 elevated and 14 decreased serum proteins in the lymphoma group relative to the healthy group. Six candidate increased proteins in canine lymphomas were beta-actin cytoplasmic 1 (ACTB, p=0.04), haptoglobin (p=0.002), beta-2 microglobulin (aaaaaaaa2M, p=0.007), beta-2 glycoprotein 1 (APOH, p=0.03), metalloproteinase inhibitor 1 (TIMP-1, p=0.03), and CD44 antigen (p=0.02). When compared between B- and T-cell lymphomas, B-cell phenotypes had upregulated immunoglobulin (Ig) heavy chain V region GOM (p=0.02), clusterin (p=0.01), apolipoprotein C1 (APOC1, p=0.05), and plasminogen (p=0.02).Conclusion:These findings were investigated quantitative serum proteomes between B- and T-cell lymphomas using TMT-based MS. ACTB, aaaaaaaa2M, APOH, TIMP-1, CD44 antigen, Ig heavy chain V region GOM, and APOC1 are novel candidate proteins and might serve as a lymphoma biomarker in dogs. However, evaluation with an increased sample size is needed to confirm their diagnostic and prognostic ability.

Genes Encoding the Glycoprotein Hormone GPA2/GPB5 and the Receptor LGR1 in a Female Prawn.

In vertebrate reproduction, metabolism, growth and development, essential roles are played by glycoprotein hormones, such as follicle-stimulating hormone (FSH), luteinizing hormone (LH) and thyroid-stimulating hormone (TSH), all of which are heterodimers consisting of two subunits, a structurally identical alpha subunit, and a variable beta subunit, which provides specificity. A 'new' glycoprotein hormone heterodimer identified in both vertebrates and invertebrates, including decapod crustaceans, was shown to be composed of the glycoprotein alpha 2 (GPA2) and glycoprotein beta 5 (GPB5) subunits. The putative receptor for GPA2/GPB5 in invertebrates is the leucine-rich repeat-containing G protein-coupled receptor 1 (LGR1). In this study in the giant freshwater prawn, Macrobrachium rosenbergii, we identified and characterized the GPA2 (MrGPA2), GPB5 (MrGPB5) and LGR1 (MrLGR1) encoding genes and revealed their spatial expression patterns in female animals. Loss-of-function RNA interference (RNAi) experiments in M. rosenbergii females demonstrated a negative correlation between MrGPA2/MrGPB5 silencing and MrLGR1 transcript levels, suggesting a possible ligand–receptor interaction. The relative transcript levels of M. rosenbergii vitellogenin (MrVg) in the hepatopancreas were significantly reduced following MrGPA2/MrGPB5 knockdown. MrLGR1 loss-of-function induced MrVg receptor (MrVgR) transcript levels in the ovary and resulted in significantly larger oocytes in the silenced group compared to the control group. Our results provide insight into the possible role of GPA2/GPB5-LGR1 in female reproduction, as shown by its effect on MrVg and MrVgR expression and on the oocyte development. Here, we suggest that the GPA2/GPB5 heterodimer act as a gonad inhibiting factor in the eyestalk-hepatopancreas-ovary endocrine axis in M. rosenbergii.

Anti-Nuclear Antibodies Patterns in Patients With Systemic Lupus Erythematosus and Their Correlation With Other Diagnostic Immunological Parameters.

BackgroundAntinuclear antibodies (ANA) are major immunodiagnostic tools in systemic lupus erythematosus (SLE); however, their clinical and pathogenic roles are not yet elucidated and are a subject of controversy.ObjectivesThe aim of the study is to explore the pathogenic significance of ANA patterns among SLE patients, by analyzing their association with ANA titers, complement levels and other pathogenic immune markers, namely, anti-double-stranded DNA (anti-dsDNA), complements C3 and C4, rheumatoid factor (RF), anticardiolipin antibodies IgG (ACL IgG) and IgM (ACL IgM), Beta-2 Glycoprotein 1 Antibodies (β2-GP) IgG (β2-IgM) and IgM (β2-IgM), and lupus anticoagulant (LA).MethodA comparative cross-sectional study was conducted among 495 SLE patients, who were diagnosed and classified by consultant rheumatologists according to the new European League Against Rheumatism (EULAR)/American College of Rheumatology (ACR) 2019 criteria. SLE immunodiagnostic profiles were analyzed including the following parameters: ANA antibody titers and staining patterns, anti-dsDNA, C3 and C4 levels, aCL, and anti-β2-GP and LA.ResultThe most frequently observed ANA patterns were the speckled (52.1%) and homogeneous (35.2%) patterns, while other patterns were rare representing less than 7% of the patients each. ANA titers were highest in patients with mixed pattern followed by the speckled pattern. Of all the investigated patterns, the peripheral pattern showed the most pathogenic immune profile, namely, highest levels of anti-dsDNA, lowest levels of C4, and highest levels of aCL and β2-GP IgG and IgM.ConclusionThis retrospective study showed that speckled followed by homogeneous ANA patterns were predominant accounting for 52.1 and 35.2% of the patients. The ANA pattern showed several associations with other immune markers that are documented to have significant clinical implications in SLE. Peripheral, mixed, and speckled patterns were associated with higher profiles of immune markers indicative of a potential prognostic value of these patterns in SLE.

IgA anti-beta-2 glycoprotein I antibodies in chronic hepatitis C

Background and study aimsAntiphospholipid antibodies (aPL) have been reported not only in various autoimmune conditions but also in other infections, such as chronic hepatitis C (CHC) infection. The aim of this study is to evaluate the frequency of aPL in patients with CHC. Patients and methodsNinety-six CHC patients and 90 healthy blood donors (HBD) were studied. Fifty-three of the patients were under treatment, and 43 had not yet received any treatment. IgG, IgA, and IgM antibodies against cardiolipin (aCL) and beta-2 glycoprotein I (aβ2GPI) were detected by ELISA. ResultsWe found that the frequency of aPL (aCL and/or aβ2GPI) was significantly higher in CHC patients than in controls (51% vs 11.1%, p <10−6). The frequencies of aCL and aβ2GPI were significantly higher in patients than in HBD (27.1% vs 5.5%, p < 10−3, and 44.8% vs 11.1%, p < 10−6, respectively). The isotype distribution of aCL and aβ2GPI demonstrated that aCL-IgG and aβ2GPI-IgA were more frequent in patients than in healthy subjects (21.9% vs 2.2%, p < 10−3, and 38.5% vs 7.8%, p < 10−6, respectively). In CHC patients, the frequency of aβ2GPI was significantly higher than that of aCL (44.8% vs 27.1%, p = 0.01). aβ2GPI-IgA was significantly more frequent than aβ2GPI-IgG (38.5% vs 7.3%, p <10−6), aβ2GPI-IgM (38.5% vs 9.4%, p <10−3), and aCL-IgG (38.5% vs 21.9%, p = 0.01). No difference in aPL frequency was observed between the treated and untreated patients. ConclusionOn the basis of the findings of this study, aPL, particularly aβ2GPI-IgA and aCL-IgG, are frequent in CHC patients.

Analysis of the Relationship between the Expression Level of TTR and APOH and Prognosis in Patients with Colorectal Cancer Metastasis Based on Bioinformatics.

The expression of TTR and apolipoprotein H (APOH) genes and their relationship with prognosis in patients with colorectal cancer (CRC) metastasis by using bioinformatics analysis techniques are explored. The expression profiles of related genes in patients with CRC metastasis are retrieved from the Gene Expression Omnibus (GEO) database. The core genes transthyretin (TTR) and APOH are screened by constructing protein-protein interaction (PPI) network, and the corresponding patient data of 327 patients are extracted and included in the metastasis group. The TTR and APOH genes of 300 patients without CRC metastasis are screened and included in the control group. The relationship between the expression levels of TTR and APOH and the clinicopathological parameters of patients with CRC metastasis is analyzed. Kaplan–Meier survival curve is drawn to observe the influence of overexpression and low expression of TTR and APOH on the prognosis and survival of patients in the metastatic group. Receiver operating characteristic (ROC) curve is drawn to observe the prognostic efficacy of combined TTR and APOH detection in patients with CRC metastasis. The experimental results show that bioassay can confirm the close relationship between TTR, APOH, and patients with CRC metastasis. Regular detection of serum TTR and APOH expression can effectively assess the patient's condition and take measures to improve the prognosis of the patients.

Very Early Onset Severe Preeclampsia and Eclampsia Syndrome with Posterior Reversible Encephalopathy Syndrome- A Catastrophe

Early onset severe preeclampsia and eclampsia is a direct cause in the deadly triad of maternal mortality along with haemorrhage and sepsis with high perinatal mortality. Antiphospholipid Syndrome (APS) with Recurrent Pregnancy Loss (RPL) may also result in early onset severe preeclampsia, eclampsia and its catastrophic complications. Present report is a rarest case of a 33-year- old, unbooked G6A5 at 21+4 weeks gestation with eclampsia, HELLP (Haemolysis, Elevated liver Enzymes, Low Platelet count) syndrome, Posterior Reversible Encephalopathy Syndrome (PRES), Intrauterine foetal demise (IUFD) with RPL due to APS with Hepatitis B positive status. Patient was managed by a multidisciplinary team of Obstetricians, Intensivists, Cardiologists and Neurologists. Eclampsia was managed with magnesium sulphate (Pritchard’s regime), intravenous labetalol and delivery of stillborn male foetus weighing 645 gm achieved within 24 hours of eclamptic seizure. Lupus anticoagulant and beta-2 glycoprotein IgM (Immnoglobulin M) antibodies were positive in high titres confirming APS. Postdelivery patient had persistent headache with blurring of vision. A diagnosis of PRES was confirmed on Magnetic Resonance Imaging (MRI) brain. Patient was put on Levetiracetam and low molecular weight heparin and was discharged in satisfactory condition on seventh postpartum day with an advice to follow- up at two weeks, six weeks and 12 weeks. Early onset preeclampsia is encountered in Obstetrics but very early onset severe preeclampsia (&lt;24 weeks of gestation) and eclampsia with HELLP syndrome, PRES in a patient of RPL due to APS is rarest. Early diagnosis of APS and early onset preeclampsia with timely referral to tertiary care and multidisciplinary management can save women from severe dreaded complications with a good obstetric outcome in future.

Ordering Patterns of Thrombophilia Testing and Its Appropriateness

BackgroundDespite society recommendations to limit thrombophilia testing, this testing is often sent inappropriately. The results of thrombophilia testing frequently do not affect management. Additionally, interpretation of thrombophilia testing is confounded by acute thrombosis, anticoagulation (AC) therapy, and other medical comorbidities. Incorrect test selection is also a source of unnecessary testing [i.e. Factor V (FV) activity level instead of Factor V Leiden (FVL)].The aim of our study was to assess ordering patterns for thrombophilia testing by qualifying the number of tests, identifying the requesting services, and assessing the appropriateness of testing.MethodsThis was a retrospective study of thrombophilia testing performed at LAC+USC Medical Center, Los Angeles, CA from January 1, 2019 to December 31, 2019. A laboratory query of thrombophilia testing was performed to identify eligible adult patients who received thrombophilia testing without a prior confirmed thrombophilia. Thrombophilia testing included: FVL and prothrombin 20210 gene mutations; activated protein C (APC) resistance; antithrombin, or protein C or S activity levels; and antiphospholipid syndrome (APS) evaluation with lupus anticoagulant, cardiolipin (CL) immunoglobulins IgM/G, and beta-2 glycoprotein (b2gp) IgM/IgG; and JAK2 V617F mutation. Homocysteine (HC) levels and methylenetetrahydrofolate reductase (MTHFR) gene mutation testing were considered to have limited clinical utility. FV activity and phosphatidylserine IgM/IgG were considered incorrect tests.The electronic medical record was reviewed for clinical history, indication for testing, requesting service, and appropriateness. The criteria for defining appropriateness were determined based on major society guidelines and literature review. The main criteria are summarized here. Testing was considered inappropriate for a provoked venous thromboembolism (VTE) or stroke/transient ischemic attack (TIA). For unprovoked VTE, testing was considered inappropriate for patients > 45 yo except for APS testing. For non-stroke arterial thrombosis, recurrent pregnancy loss or stillbirth, and diagnostic evaluation of suspected lupus, APS testing only was considered appropriate.Results450 patients underwent thrombophilia testing with a mean age of 42 (range: 18-90); 76% were female and 81% were Hispanic.A total of 1698 thrombophilia tests were sent by 27 services. Testing was done in the following settings: inpatient (40%), outpatient (59%), and emergency department (1%). The mean tests per patient were 3.7 (range: 1-12). The most common requesting services were rheumatology (24%), obstetrics-gynecology (19%), and internal medicine/medicine-pediatrics (14%). Hematology requested 10% of tests.Common indications for testing were VTE (21%), rheumatology-related (25%), pregnancy-related (13%), ischemic stroke/TIA (9%), ocular-related (7%), non-stroke arterial thrombosis (3%), and dermatology-related (4%) (Table 1). 5% (84 tests) were sent for the evaluation of other non-thrombotic conditions. 8% (132 tests) were sent for > 1 indication, such as concurrent arterial and venous events.840 tests (49%) were deemed inappropriate. Common reasons for inappropriate testing included provoked VTE events, stroke/TIA, APS testing after first pregnancy loss, current AC, and duplicate testing (Table 2). APS testing issues included testing for LAC while on AC, incomplete testing (both CL and b2GP not sent), incorrect tests (phosphatidylserine IgM/IgG), and repeat testing < 12 weeks from prior. Incorrect/redundant testing for FVL included: FV activity levels (36 total tests; 9 ordered in additional to FVL and 27 ordered instead of FVL) and APC resistance ordered simultaneously with FVL in 7 patients. Of note, 92 tests were sent for evaluation of non-thrombotic conditions.ConclusionsThrombophilia testing is often done inappropriately. Correct test selection is also a relatively common issue, particularly with APS and FVL testing. Given the large number of services ordering thrombophilia testing at our training hospital and this testing being sent for a variety of reasons, it is unlikely that physician education alone will lead to a substantial or sustained decrease in the number of inappropriate tests. Rather, it may be necessary to restrict at least some thrombophilia testing to certain services. [Display omitted] DisclosuresPiatek: Rigel: Consultancy, Research Funding; Alexion: Consultancy, Research Funding; Apellis: Research Funding; Dova: Consultancy, Speakers Bureau.

Laboratory Analysis of Antiphospholipid Antibodies with Clinical Correlation: A Hospital-Based 10 Year Study

Abstract Introduction/Objective Antiphospholipid syndrome (APS) is an autoimmune disorder and its diagnosis requires both laboratory evaluation of antiphospholipid antibodies (APAs) and clinical correlation. This study focused on the laboratory analysis of APS with clinical correlation in a hospital-based patient population. Methods/Case Report From 2010-2020, immunological studies for APAs were performed at Department of Pathology, William Beaumont Hospital – Troy, Michigan and patients with positive results were selected for further analysis. Four APAs were included: IgG and IgM beta-2 glycoprotein I antibodies (IgG B2GA and IgM B2GA), and IgG and IgM anticardiolipin antibodies (IgG ACA and IgM ACA). Clinical information was collected from hospital electronic medical chart with focus on thromboembolism and abortion history, autoimmune disorders, antithrombotic therapies and other studies. Results (if a Case Study enter NA) There were 167 patients, 136 females and 31 males (F/M=4.4). The median age for females was 50 and for males, 57. Overall, B2GA was present more than ACA (102/81) in females, and ACA was present slightly more than B2GA (20/17) in males. IgM APAs were much more common than IgG APAs; IgM B2GA/IgG B2GA: 87/32 (p&amp;lt;0.01) and IgM ACA/IgG ACA: 81/20 (p&amp;lt;0.01). Most APAs were at the low-intermediate titer range and increased APAs titer correlated with decreased positive rate: APAs titer at 20-40, 41-80 and &amp;gt;80 with positive results at 115, 61 and 44 respectively. In addition to documented autoimmune disorders, pregnancy miscarriage was a common clinical condition (24/66) in females at age &amp;lt;45. But for females at age &amp;gt;45, vascular thrombosis became more common (18/70). Vascular thrombosis was a common clinical disease in males (16/31) followed by autoimmune disorders (10/31). Most patients with vascular thrombosis received antithrombotic therapies. Conclusion Female patients positive for APAs was significantly more and younger than males in this study. IgM B2GA was the most common type of APAs followed by IgM ACA. IgG B2GA and IgG ACA were much less common. In younger female patients autoimmune disorders and abortion were the most common clinical conditions. In older female and male patients, autoimmune disorders and vascular thrombosis were the most common clinical conditions. The titer of APAs detected was at low-intermediate range in most patients. The relationship between the titer of APAs and clinical conditions needs further study.