Lambda Chains Research Articles

Structural modification of 2-phenylquinoline-4-carboxylic acid containing SIRT3 inhibitors for the cancer differentiation therapy.

Inhibition of SIRT3 exhibited potency in triggering leukemic cell differentiation. In discovery of potent SIRT3 inhibitors for cancer differentiation therapy, structural modification was performed on the previously developed lead compound P6. A total of 33 compounds were designed and synthesized. In the enzyme inhibitory assay, several molecules S18, S26, S27 and T5 showed potent SIRT3 inhibitory activity with IC50 value of 0.53, 1.86, 5.06, and 2.88 μM, respectively. Moreover, the tested compounds exhibited SIRT3 inhibitory selectivity over SIRT1 and SIRT2. Compounds S27 and T5 were potent in inhibition the growth of MM1.S and RPMI-8226 cells in the invitro antiproliferative test. Significantly, representative compounds, especially S27 and T5, promoted differentiation of tested MM cells in the cellular morphological evaluation, accompanied by increasing the expression of differentiation antigen CD49e and human immunoglobulin light chain lambda and kappa. Additionally, molecule S18 without antiproliferative potency itself, showed significant inhibitory activity against growth factor IL-6 induced RPMI-8226 cell proliferation. Collectively, potent SIRT3 selective inhibitors with MM cell differentiation potency were developed for further discovery of anticancer drugs.

Primary diffuse large B-cell lymphoma of the central nervous system identified with CSF biomarkers

BackgroundDiagnosis of primary diffuse large B-cell lymphoma of the central nervous system (PCNSL) is challenging and often delayed. MRI imaging, CSF cytology and flow cytometry have a low sensitivity and even brain biopsies can be misleading. We report three cases of PCNSL with various clinical presentation and radiological findings where the diagnosis was suggested by novel CSF biomarkers and subsequently confirmed by brain biopsy or autopsy.Case presentations.The first case is a 79-year-old man with severe neurocognitive dysfunction and static ataxia evolving over 5 months. Brain MRI revealed a nodular ventriculitis. An open brain biopsy was inconclusive. The second case is a 60-year-old woman with progressive sensory symptoms in all four limbs, evolving over 1 year. Brain and spinal MRI revealed asymmetric T2 hyperintensities of the corpus callosum, corona radiata and corticospinal tracts. The third case is a 72-year-old man recently diagnosed with primary vitreoretinal lymphoma of the right eye. A follow-up brain MRI performed 4 months after symptom onset revealed a T2 hyperintense fronto-sagittal lesion, with gadolinium uptake and perilesional edema. In all three cases, CSF flow cytometry and cytology were negative. Mutation analysis on the CSF (either by digital PCR or by next generation sequencing) identified the MYD88 L265P hotspot mutation in all three cases. A B-cell clonality study, performed in case 1 and 2, identified a monoclonal rearrangement of the immunoglobulin light chain lambda (IGL) and kappa (IGK) gene. CSF CXCL-13 and IL-10 levels were high in all three cases, and IL-10/IL-6 ratio was high in two. Diagnosis of PCNSL was later confirmed by autopsy in case 1, and by brain biopsy in case 2 and 3.ConclusionsTaken together, 5 CSF biomarkers (IL-10, IL-10/IL-6 ratio, CXCL13, MYD88 mutation and monoclonal IG gene rearrangements) were strongly indicative of a PCNSL. Using innovative CSF biomarkers can be sensitive and complementary to traditional CSF analysis and brain biopsy in the diagnosis of PCNSL, potentially allowing for earlier diagnosis and treatment.

#2282 Outcomes of renal amyloidosis: 8 years experience of tertiary center

Abstract Background and Aims Amyloidosis is a disease caused by the extracellular deposition of amyloid material that leads to organ dysfunction. Renal involvement is frequent and is characterized by amyloid fibrils pathologic deposition in glomeruli, vessels and interstitium. The most common presentation is nephrotic-range proteinuria or nephrotic syndrome and can progress to end stage kidney disease (ESKD). Prognosis is usually poor, especially with cardiac and severe renal involvement that requires renal replacement therapy. The goal of this study was to analyse the outcomes of patients with amyloidosis followed at the nephrology clinic in our centre. Method Descriptive analysis of all confirmed amyloid patients, followed by the Nephrology department from January 2015 to June 2023. Results We identified 49 patients with a mean age of 64 ± 16 years [18-88], mainly Caucasian (n = 47, 82%) and male (n = 28, 57%). The majority of patients were diagnosed through renal biopsy (26, 53%). The most common amyloid type was AL (n = 18, 38%), followed by ATTR (n = 16, 33%) and AA (n = 12, 25%). In 3 patients the amyloid type couldn't be characterized. Renal presentation was mostly as CKD 28/49 (57%), 15/49 (30.6%) as nephrotic syndrome and 6/49 (12%) patients as AKI. At presentation, almost half of the patients (24/49, 49%) had nephrotic-range proteinuria, with a mean proteinuria of 6.72 ± 6.12 g/d [0.103-19.8] and mean sCr was 2.6 ± 3.1 mg/dl [0.5-18.9]. AL amyloidosis had an almost equal distribution between lambda (9/18, 50%) and kappa chain (8/18, 44%, one unknown). Extrarenal involvement was frequent, mainly cardiac (12/18, 67%). Also 12/18 (67%) patients needed dialysis after a mean of 9 ± 15 months post-diagnosis, 16/18 (89%) received treatment and mortality was 61%, in mean 16 ± 17 months post-diagnosis, mainly due to disease progression. AA amyloidosis was secondary to chronic infection in 7/12 (58%), 3/12 (25%) autoimmune diseases and in 2/12 (17%) unknown. Extrarenal involvement was less frequent, also mainly cardiac (3/12, 25%). 7/12 (58%) patients needed dialysis after a mean of 97 ± 117 months post-diagnosis and mortality was 58%, in mean 46 ± 66 months post-diagnosis, mainly due to acute infection. Two patients were diagnosed already on dialysis. Regarding ATTR amyloidosis, all patients had a positive genetic test, extrarenal involvement was very frequent (13/16, 81%), mostly cardiac and peripheric nervous system. No patients required dialysis during follow-up, and one patient died of unknown cause. Conclusion In our cohort, renal AL and AA amyloidosis had poor prognosis, with a high progression to ESKD and very significant mortality, irrespective of treatment. Conversely, ATTR amyloidosis patients had lower progression to dialysis and death, probably as a consequence of disease-modifying therapies instituted in recent years.

#2963 Light chain deposition in the kidney is more frequent in patients with proliferative lupus nephritis

Abstract Background and Aims Light chain deposition in the kidney is an important histologic feature having a significant role in the pathogenesis of glomerular and autoimmune diseases, including lupus nephritis (LN). However, there is no study examining differences in light chain deposition profile between proliferative and non-proliferative LN. Method We have conducted a retrospective cohort study to evaluate the characteristics and prognostic significance of light chain deposition profile in the kidney of subjects with proliferative and non-proliferative LN. We have collected data on demographics, clinical and laboratory parameters and histopathology (light, immunofluorescent and electron microscopy). Lambda domination (LD) was defined as lambda intensity—kappa intensity ≥ +1. SLE was diagnosed using the American College of Rheumatology criteria and renal outcomes per KDIGO guidelines. Proliferative LN was defined as those containing classes III and IV, while all other classes comprised non-proliferative LN. Results A total of 56 patients with biopsy-proven LN were followed up for at least one year after kidney biopsy (79% women, mean age at biopsy 38 ± 13 years). A total of 71% of patients had proliferative LN. Mean number of glomeruli per biopsy sample was 26 ± 12. Mean immunofluorescent intensity was 1.6 ± 1.0 for lambda and 1.8 ± 1.0 for kappa light chain. A total of 42 (75%) patients had light chain deposition in the glomerulus with 4 (7%) having restricted lambda chain deposition and none had restricted kappa chain deposition. Patients with proliferative LN had more frequent light chain deposition in glomeruli than those with non-proliferative lupus (88% vs. 57%, p = 0.049) , with no difference in LD (24% vs. 14%, p = 0.71). There was no difference between frequency of restricted lambda chain deposition between proliferative and non-proliferative LN (5% vs. 17%, p = 0.60). A total of 12 (21%) patients had LD in the glomerulus. When examining renal outcomes at one year post-biopsy, 55% of patients achieved complete response (CR), 30% achieved partial response (PR) and 15% had no response. There were no differences in achievement of remission (CR or PR) between patients with proliferative and non-proliferative LN (87% vs. 90%, p = 0.93). We also found no difference in frequency of light chain deposition and achievement of remission in proliferative or non-proliferative LN (both p>0.05). Conclusion Light chain deposition is prevalent in LN and is more frequent in patients with proliferative LN. Further studies examining their pathophysiologic properties and potential prognostic value are much warranted.

Elimination of Medium/High Molecular Weight Solutes. Comparison of High Flow Hemodialysis with Extended Hemodialysis

Introduction: Post-dilution online hemodiafiltration is the most efficient extracorporeal depurative treatment of CKD. Recently a new type of membrane has been developed, with a higher cut-off point also called a medium cut-off point, which has the capacity to eliminate higher molecular weight molecules in hemodialysis. The hemodialysis performed with these membranes has been called “Expanded Hemodialysis”. Objective: Compare the purifying efficacy of medium and high molecular weight molecules in patients dialyzed with high-flux hemodialysis, VitaPES 210HF, and with patients treated with expanded hemodialysis with the medium cut-off dialyzer, Elisio-HX. We also assessed the effect that the increased removal of inflammatory mediators by MCO hemodialysis had on fecal Calprotectin levels. Patients and methods: This is a prospective observational cross-over study in which 8 prevalent hemodialysis patients were followed for two months. Blood levels of IL-6, C-reactive protein (CRP), β2-microglobulin, Kappa and Lambda immunoglobulin light chains, and serum albumin were determined before and after each hemodialysis session. Results: The percentage of reduction of medium and higher molecular weight molecules: β2microglobulin, IL-6, Kappa and Lambda chains and CRP were higher with the Elisio-21HX dialyzer compared to the VitaPES 210HF dialyzer. There was no difference in albumin clearance between the two dialyzers. Fecal calprotectin levels were lower in patients dialyzed with Elisio-21HX. Conclusion: The medium cutoff dialyzer, Elisio-HX, is more efficient in the elimination of medium/high molecular weight molecules than the VitaPES HF high-flux dialyzer, with the same albumin elimination. Improving inflammation at the local intestinal level with lower levels of fecal Calprotectin.

Comprehensive Stress Stability Studies Reveal the Prominent Stability of the Liquid-Formulated Biotherapeutic Asymmetric Monovalent Bispecific IgG1 Monoclonal Antibody Format

The developed asymmetric monovalent bispecific IgG1 or Duet monoclonal antibody (Duet mAb) has two distinct fragment antigen-binding region (Fab) subunits that target two different epitope specificities sequentially or simultaneously. The design features include unique engineered disulfide bridges, knob-into-hole mutations, and kappa and lambda chains to produce Duet mAbs. These make it structurally and functionally complex, so one expects challenging developability linked to instability, degradation of products and pathways, and limited reports available. Here, we have treated the product with different sources of extreme stress over a lengthy period, including varying heat, pH, photo stress, chemical oxidative stress, accelerated stress in physiological conditions, and forced glycation conditions. The effects of different stress conditions on the product were assessed using various analytical characterization tools to measure product-related substances, post-translational modifications (PTMs), structural integrity, higher-order disulfide linkages, and biological activity. The results revealed degradation products and pathways of Duet mAb. A moderate increase in size, charge, and hydrophobic variants, PTMs, including deamidation, oxidation, isomerization, and glycation were observed, with most conditions exhibiting biological activity. In addition, the characterization of fractionated charge variants, including deamidated species, showed satisfactory biological activity. This study demonstrated the prominent stability of the Duet mAb format comparable to most marketed mAbs.

Phenotypes and outcome of diffuse pulmonary non-amyloid light chain deposition disease.

Light chain deposition disease (LCDD) is a very rare entity. Clinical manifestations of LCDD vary according to the organs involved. Data on pulmonary LCDD are scarce and limited to small series or case reports. This study aimed to describe the characteristics and outcome of diffuse pulmonary non-amyloid LCDD localized to the lungs. A multicenter retrospective cohort study was conducted. Clinical characteristics were collected, and chest CTs were centrally reviewed. The diagnosis of pulmonary non-amyloid LCDD was confirmed by immunohistochemistry. Thirty-one cases were identified (68% female), with a median age at diagnosis of 50 years (IQR 20). Baseline FEV1/FVC was < 0.70 in 45% of patients. Mean (± SD) FEV1 and DLCO were 86% ± 26.2 and 52% ± 23.9, respectively. CT revealed peculiar patterns of thin-walled cysts (58%) and thin-walled cystic bronchiectases (27%). Increased serum kappa light chain was found in 87% of patients. Histological analysis showed kappa light chain deposits in all patients, except one with lambda chain deposits. Median annual FEV1 decline was 127 ml (IQR 178) and median DLCO decline was 4.3% (IQR 4.3). Sixteen patients received immunomodulatory treatment or chemotherapy; serum light chain levels decreased in 9 cases (75%), without significant improvement in FEV1 (p = 0.173). Overall, 48% of patients underwent bilateral lung transplantation. Transplant-free survival at 5 and 10 years were 70% and 30%, respectively. An annual FEV1 decline greater than 127 ml/year was associated with increased risk of death or transplantation (p = 0.005). Diffuse pulmonary LCDD is characterised by female predominance, a peculiar imaging pattern with bronchiectasis and/or cysts, progressive airway obstruction and severe DLCO impairment, and poor outcome. Lung transplantation is a treatment of choice.

Morphological findings in different subtypes of hepatic amyloid

The classic findings have been well described for light-chain amyloid involving the liver. In addition to light chain, however, many additional proteins are now known to be amyloidogenic and can involve the liver. A total of 58 surgical pathology specimens with amyloid deposits were analyzed for patterns of amyloid deposition, including amyloid from light chain lambda (N = 17), light chain kappa (N = 15), transthyretin (N = 15), serum amyloid A (N = 4), apolipoprotein A1 (N = 4), fibrinogen alpha (N = 2), LECT2 (N = 1). Amyloid deposits predominately targeted the liver vasculature, including the walls of the hepatic arteries, portal veins, and sinusoids. While there was overlap, light chain amyloid predominately involved the sinusoids, while transthyretin amyloid predominately targeted the hepatic arteries, especially the larger ones in the hilum and larger portal tracts. Serum amyloid A formed nodular deposits that started in the portal vasculature but then extended into the portal tract stroma, leading to large, bulbous, portal-based amyloid deposits. Apolipoprotein A amyloid also formed large portal-based nodules. Fibrinogen was mild and subtle on H&E and predominately affected portal veins. Amyloid deposits in hilar nerves were prominent with amyloid light chain, transthyretin, and apolipoprotein A1. In conclusion, the histology of hepatic amyloid is diverse and shows several distinct clusters of findings that can aide in recognition in surgical pathology specimens.

P807 HLA type impact on immunization and efficacy of ustekinumab in IBD patients: a multicenter study

Abstract Background About 30% of patients treated with ustekinumab will present primary or secondary failure. Outcomes and frequency of ustekinumab immunization are still controversial. Some HLA types (especially DQA1*05) appear to be predictive factors of anti TNFα immunization. This study aimed to assess the impact of HLA type on the risk of immunization and on the efficacy of ustekinumab. Methods IBD patients treatede with ustekinumab in Saint-Etienne and Lyon hospitals were included between 2017 and 2023. Ustekinulab through level was measured by ELISA and HLA type (DQA1, DQB1, DRB1) determined by PCR. Anti-drug antibodies (ADA) detection was done using both a "drug sensitive" assay (ELISA) and an innovative "drug tolerant" method based on anti human lambda chain. Data about clinical, biological (CRP, fecal calprotectin) and endoscopic responses were retrospectively collected. Results Among the 115 patients included (43% males, mean age of 40 years old), 98 had Crohn disease. Nighty-eight percent of the patients were previously treated with at least one anti-TNFα and 33% of them were immunized. In this study, 43 patients were immunized against ustekinumab with the "drug tolerant" assay and only 2 with the "drug sensitive" method. Immunization tended to reduce (not significatively) clinical, biological and endoscopic responses. Ustekinumab through level was significately higher for patients with good clinical (p = 0.009) and biological (p = 0.036). HLA DQA1*05 or DRB1*03 carriage was associated with higher risk of immunization (OR = 5.13, p = 0.048 / OR = 5.36, p = 0.006) and with a trend to decrease clinico-biological response. Other HLA types, notable DQA1*01 tended to be associated with a lower immunization (OR = 0.7, p=0.3) and a better response (OR = 2.55, p = 0.018). A small group involved (26 patients), combination therapy at induction had no significant benefit in terms of immunization prevention or therapeutic response. Sub-groups analysis seemed to confirm some HLA types involvement, by breaking free of imbalances due to haplotype transmission. Conclusion Our study suggests the usefulness of ustekinumab monitoring and a more common immunization than previously described. HLA type seem to influence immunization and ustekinumab response. Some specific HLA types and patients profiles tend to be associated with a higher risk of immunization not only to anti-TNFα but also to ustekinumab.

Sensitization to avian and fungal proteins in different work environments

IntroductionHypersensitivity pneumonitis (HP) is usually caused by the inhalation of avian and fungal proteins. The present study assesses a cohort of Urban Pest Surveillance and Control Service (UPSCS) workers with high exposure to avian and fungal antigens, in order to identify their degree of sensitization and the potential risk of developing HP.MethodsWorkers were divided according to their work activity into Nest pruners (Group 1) and Others (Group 2). All individuals underwent a medical interview, pulmonary function tests and the determination of specific IgG antibodies. Antigenic proteins of pigeon sera were analysed using two-dimensional immunoblotting. Proteins of interest were sequenced by liquid-chromatography–mass spectrometry (LC–MS).Results101 workers were recruited (76 men, average age: 42 yrs); (Group 1 = 41, Group 2 = 60). Up to 30% of the study population exhibited increased levels of IgGs to pigeon, small parrot and parrot, and up to 60% showed high levels of Aspergillus and Penicillium IgGs. In Group 1, specific parakeet and Mucor IgGs were higher (p = 0.044 and 0.003 respectively) while DLCO/VA% were lower (p = 0.008) than in Group 2. Two-dimensional immunoblotting showed protein bands of 20–30 KDa recognized by HP patients but not by workers. LC–MS analysis identified Ig Lambda chain and Apolipoprotein A-I as candidate proteins for distinguishing HP patients from exposed workers.ConclusionsTwo pigeon proteins were identified that may play a role in the development of pathological differences between HP patients and exposed workers. DLCO/VA may have a predictive value in the development of HP disease.

Pathomorphological Features of the Novel Coronavirus Disease in Patients with Systemic Amyloidosis.

Amyloidosis is one of the rare systemic illnesses characterized by the deposition of amyloid fibrils in various organs and tissues. There is a common point between COVID-19 and systemic amyloidosis regarding the multiorgan involvement in the pathological process which leads to a heightened risk for severe morbidity and mortality in amyloidosis patients who contracted COVID-19. We performed a pathomorphological analysis of the autopsy records of 22 patients who had COVID-19 and pre-existing systemic amyloidosis. The premortem diagnosis of systemic amyloidosis was established in 55% of patients, and in other 45% of cases, amyloidosis was found at autopsy. Based on the results of immunohistochemical amyloid typing, amyloid A (AA) amyloidosis was detected in 23%, amyloid light chain (AL) lambda in 32%, AL kappa-in 9%, and transthyretin (ATTR) amyloidosis-in 36% of observations. Immunohistochemical staining with an antibody against SARS-CoV-2 Spike (S) protein revealed positive immune reactions in type II alveolocytes in 59% of deceased persons. The analysis of autopsy findings indicates that patients with systemic amyloidosis are more likely to experience an aggressive clinical course of COVID-19 which leads to a multiorgan failure and a higher risk of fatal outcome.

Tear film proteome changes following Tobradex® therapy in anterior blepharitis.

The management of blepharitis continues to challenge clinicians due to the poorly understood aetiology of the condition. We recently identified the family of intracellular plakin proteins as essential driving forces underlying anterior blepharitis. A large-scale protein analysis was used to study if a topical dexamethasone/tobramycin solution could be used to reverse the expression of plakin proteins. Tear film samples from treatment naïve patients with anterior blepharitis (n = 15) were collected with Schirmer filtration paper. A subgroup of the patients (n = 10) received treatment with a dexamethasone/tobramycin 1 + 3 mg/mL ophthalmic suspension (Tobradex®) for 3 weeks and collection of tear film samples was repeated. The samples were analysed with label-free quantification nano liquid chromatography-tandem mass spectrometry requiring quantification in at least 70% of the samples in each group. Proteins were considered differentially expressed if p < 0.05. Following Tobradex® intervention, 27 proteins were upregulated while 61 proteins were downregulated. Regulated proteins after Tobradex® treatment were involved in intermediate filament cytoskeleton organization including downregulation of the plakin proteins envoplakin, epiplakin and periplakin. Plectin, a protein of the plakin family, remained unchanged after Tobradex® therapy. Tobradex® treatment resulted in the regulation of proteins involved in translation including a cluster of downregulated ribosomal proteins. Tobradex® intervention was associated with the regulation of proteins involved in fructose metabolism and glycolytic processes including fructose-1.6-bisphosphatase 1, fructose-bisphosphate aldolases A and B, pyruvate kinase PKM and transketolase. Ig lambda chain V-I region, prominin-1, and protein Niban were upregulated after Tobradex® treatment. Tobradex treatment reversed the expression of plakin proteins in anterior blepharitis. Topical solutions which inhibit the expression of plakin proteins may have the potential to restore the ocular surface integrity in anterior blepharitis and should be explored further.

Атипичное течение макроглобулинемии «под маской» анемии (клиническое наблюдение)

Waldenström macroglobulinemia (WM) is a type of plasma cell disease that is a malignant B-cell lymphoplasmacytic lymphoma. In patients with WM, immunological blood tests may remain unchanged for a while without pathological changes, but then tumor infiltration of the bone marrow gradually leads to anemia and thrombocytopenia. This patient has anemic syndrome according to clinical and laboratory examination, in which a number of diseases were diagnosed. An early diagnostic method, electrophoresis allows the consideration of immunoproliferative disorders with monoclonal IgM. Immunotyping is widely used with the eye chart of gammopathies using antisera to light (G, A, M) and light (kappa, lambda) chains of immunoglobulins. An increase in blood vessels IgM &gt; 30 g/l, when identified with neurological symptoms of the source, associate these manifestations with hyperviscosity syndrome in WM.

#6658 ORAL HEALTH STATUS IN DIABETIC AND NON-DIABETIC PATIENTS ON MAINTENANCE HEMODIALYSIS TREATMENT

Abstract Background and Aims Uremic toxins and inflammation influence the oral health in patients on maintenance hemodialysis treatment. The presence of diabetes additionally aggravates the oral status. Aims: To compare the oral health status in diabetic and non-diabetic patients on different dialysis modality treatment. Method Observational, cross-section, monocentric study was conducted in 72 hemodialysis (HD) patients divided into two groups regarding the presence of Diabetes mellitus (DM). Patients were routinely designed to hemodialysis or hemodiafiltration (HDF) during the previous year of HD treatment. Demographic characteristics as patients age, dialysis vintage, laboratory inflammatory markers as C-reactive protein (CRP), albumin and Interleukin 6 (IL-6) were measured at the start of the study. Also, uremic small and middle molecules as blood urea nitrogen (BUN), creatinine, β2-microglobilin (β2M), myoglobin, albumin, free light chains kappa (FLC-k), and free light chains lambda (FLC-λ) were analyzed. Patients were examined by a dentist specialist scoring the oral hygiene index (OHI) by Greene Vermillion as good, fair and poor. Presence of tooth fillings and extractions, caries, hyperkeratosis, periodontal disease, erosions, ulceration, erythema, pigmentations, saburral tonque and uremic fetor were notified. Gingival hyperplasia (GH) was scored (1-3) with 3 for worst score and the tooth color was scored 1-3, signing 1-yellow, 2-gray, 3-white. Data was presented as mean and standard deviation for continuous and percentages for nominal values. X squared Fisher exact and Mann-Whitney test were used for statistical analysis. P&amp;lt;0.05 was considered as significant. Results The patients from group 1 - with DM (N=26) didn't differ from the non-diabetic group (N= 46) in respect of gender, age and dialysis modality but had significantly shorter dialysis vintage (48.68 ± 37.45 vs. 88.13 ± 63.29, P = .02, respectively). From the inflammatory markers only Il-6 was significantly higher in diabetics (P = .03). All the analyzed uremic toxins – small and middle molecules also didn't differ between the two groups. Diabetic patients were at 3 fold risk for manifestation of fissure, 4 fold risk for pigmentations and 7 fold risk for erythema (OR 3.58; CI:1.017-12.380, p= 0.003; OR 4.12; CI:0.684-22.870; p= 0.02, OR 4.84; CI:1.343-17.498, p= 0.000), (OR 7.25; CI:1.123-46.880, p= 0.000), respectively. GH was more likely to be present in diabetic patients (35%, 54%, 11% vs 83%, 15, 0%, p= 0.000, respectively). The presence of hyperkeratosis, periodontal disease erosions, caries, extractions, tooth fillings and the tooth colour didn't differ between the groups. Diabetics were found with higher percentage of bad oral hygiene index (38% vs 20%), but the overall comparison of OHI showed no significant difference. Conclusion Oral health is significantly deteriorated in dialysis patients, especially in those with inflammation. Diabetic patients are at higher risk of developing changes in the oral health status.

#6648 LONG TERM EFFECT OF MEDIUM CUT-OFF DIALYZER ON MIDDLE UREMIC TOXINS

Abstract Background and Aims Better removal capacities for middle and large middle molecules in hemodialysis was achieved in (HD) treatment with the new medium cut-off (MCO) membranes. The aim of this study was to evaluate the long term removal efficacy of Theranova® in standard HD in comparison with standard high-flux HD. Method 11 patients were assigned to high flux and 11 to MCO membranes in 12-months observational study. Each patient was assessed every three months from T0 with high-flux dialyzers, T1 to T5 with MCO at first, third, sixth, nineth and twelfth month by measuring pre- and post-HD samples of β2-microglobilin (β2M), myoglobin, albumin, free light chains kappa (FLC-k), and free light chains lambda (FLC-λ). The cumulative change from as RR from T0 to T5 was defined as long term outcome. Results The data showed a higher average removal rate (RR) for all the uremic toxins with Theranova® dialyzers for β2M, myoglobin, FLC-k, and FLC-λ (66.1, 53.2, 59.2.5, and 60.9%, respectively) compared to high flux (57.1, 10.8, 29.9 and 37.4%, respectively). The analyses on the cumulative RR from T0 to T5 found significant decrement of β2M, myoglobin, FLC-λ in MCO vs high flux group where those values significantly raised (14.9 vs −6.1, p = 0.043; 27.7 vs −7.8, p = 0.001; 21.9 vs −4.2, p = 0.017, respectively). The FLC-k showed borderline significantly better removal with MCO (17.8 vs 8.4. p = 0.067). The distribution of patients with 12 month middle molecules reduction was much better in the MCO group. Albumin retention was better in MCO group and did not change between T0 and T5 (p = 0.824). Conclusion Compared to high-flux membranes, MCO membranes show greater permeability for middle molecules and albumin stability in long-term report. Their use in HD standard setting is effective and safe.

Atypical Lichen Myxedematosus Associated with Multiple Myeloma

Lichen myxedema (LM) represents a group of rare skin diseases, which falls under chronic mucinoses. LM, previously described as localized scleromyxedema, however, is not associated with systemic involvement. Among the subtypes, atypical LM is rare, with few reported cases associated with multiple myeloma (MM). We present a case of MM, positive for lambda chains, with acute-onset clinical symptoms, and diagnosed with atypical LM. The patient was managed with topical corticosteroid with improvement of lesions after one month of treatment.

Astrocytes express aberrant immunoglobulins as putative gatekeeper of astrocytes to neuronal progenitor conversion

Using multi-omics analyses including RNAseq, RT-PCR, RACE-PCR, and shotgun proteomic with enrichment strategies, we demonstrated that newborn rat astrocytes produce neural immunoglobulin constant and variable heavy chains as well as light chains. However, their edification is different from the ones found in B cells and they resemble aberrant immunoglobulins observed in several cancers. Moreover, the complete enzymatic V(D)J recombination complex has also been identified in astrocytes. In addition, the constant heavy chain is also present in adult rat astrocytes, whereas in primary astrocytes from human fetus we identified constant and variable kappa chains as well as the substitution lambda chains known to be involved in pre-B cells. To gather insights into the function of these neural IgGs, CRISPR-Cas9 of IgG2B constant heavy chain encoding gene (Igh6), IgG2B overexpression, proximal labeling of rat astrocytes IgG2B and targets identification through 2D gels were performed. In Igh6 KO astrocytes, overrepresentation of factors involved in hematopoietic cells, neural stem cells, and the regulation of neuritogenesis have been identified. Moreover, overexpression of IgG2B in astrocytes induces the CRTC1-CREB-BDNF signaling pathway known to be involved in gliogenesis, whereas Igh6 KO triggers the BMP/YAP1/TEAD3 pathway activated in astrocytes dedifferentiation into neural progenitors. Proximal labeling experiments revealed that IgG2B is N-glycosylated by the OST complex, addressed to vesicle membranes containing the ATPase complex, and behaves partially like CD98hc through its association with LAT1. These experiments also suggest that proximal IgG2B-LAT1 interaction occurs concomitantly with MACO-1 and C2CD2L, at the heart of a potentially novel cell signaling platform. Finally, we demonstrated that these chains are synthesized individually and associated to recognize specific targets. Indeed, intermediate filaments Eif4a2 and Pdia6 involved in astrocyte fate constitute targets for these neural IgGs. Taken together, we hypothese that neural aberrant IgG chains may act as gatekeepers of astrocytes' fate.

Exploring the sheep (Ovis aries) immunoglobulin repertoire by next generation sequencing.

Next-generation sequencing (NGS) has revolutionized the way we determine the antibody repertoires encoded by B cells in the blood or lymphoid organs and transformed our understanding of adaptive immune responses in many species. Sheep (Ovis aries) have been widely used as a host for therapeutic antibody production since the early 1980s, however, little is known about their immune repertoires or immunological processes affecting the antibody generation. The objective of this study was to employ NGS for a comprehensive analysis of immunoglobulin heavy and light chain repertoires in four healthy sheep. We obtained >90% complete antibody sequences and nearly 130,000, 48,000 and 218,000 unique CDR3 reads for the heavy chain (IGH), kappa chain (IGK), and lambda chain (IGL) loci, respectively. Consistent with other species, we observed biased usage of germline variable (V), diversity (D) and joining (J) genes in the heavy and kappa loci, but not in the lambda loci. Moreover, the enormous diversity of CDR3 sequences was observed through sequence clustering and convergent recombination. These data will build a foundation for future studies investigating immune repertoires in health and disease as well as contribute to further refinement of ovine-derived therapeutic antibody drugs.

Difficulties in the Diagnosis of Fibrinogen Aα-Chain Amyloidosis—Literature Review and Case Report of a Patient After Kidney Transplantation

Amyloidosis is a very heterogeneous disease. Correct diagnosis is extremely important because of the various treatment options for different types of amyloidosis. This study presents a case report and literature review of the misdiagnosis of fibrinogen Aα-chain amyloidosis (AFib amyloidosis). We report a 65-year-old man diagnosed with proteinuria in 2009. The kidney biopsy revealed the presence of Congo red-stained amyloid deposits. During differential diagnosis, amyloid deposits were discovered in adipose tissue and gingiva. Bone marrow trephine biopsy showed a predominance of lambda chains presenting plasmocytes. Based on performed medical examination, light chain amyloidosis was identified. Therefore, the patient received high-dose melphalan and underwent successful autologous peripheral blood stem cell transplantation. However, proteinuria, worsening of the kidneys' function, and incorrect levels of free light chains were still observed. In 2019, due to continuous treatment failure, a previously acquired kidney biopsy was examined by mass spectrometry, and numerous fibrinogen deposits were identified. Recommended DNA analysis revealed that the patient had AFib amyloidosis. Therefore, chemotherapy treatment was abandoned, and successful kidney transplantation was performed. Today, it is essential for medical practitioners to remember the possibility of rare and hereditary types of amyloidosis. There are multiple cases where a diagnosis was wrong or delayed because of the atypical course of the disease, the coexistence of another disease, and the rarity of AFib amyloidosis, and all of these reasons may result in the wrong treatment that will delay the right therapy. However, with the new, more precise diagnostics methods, such situations will become rare.

Effects of various disaccharide adaptations on recombinant IgA1 production in CHO-K1 suspension cells.

The online version contains supplementary material available at 10.1007/s10616-023-00571-5.