Plasma exchange combined with bortezomib-based chemotherapy is effective for early renal recovery in a patient with IgD-λ type multiple myeloma.

The immunoglobulin (Ig) D type is a rare variant of multiple myeloma (MM), that accounts for 1-2% of all cases. Compared to the more common types of MM, IgD MM is known to have more severe symptoms at presentation, and a poorer prognosis. A woman was admitted to our hospital for severe acute kidney disease and disorder (AKD) and back pain, and was started on hemodialysis. The renal biopsy revealed light chain cast nephropathy. She was diagnosed with IgD-λ MM based on Bence-Jones protein expression and high IgD serum levels, and started bortezomib therapy with plasma exchange (PE). After three sessions of PE, the serum free light chain levels decreased by 92%, and she was withdrawn from dialysis. The patient underwent autologous transplantation and is still in remission, demonstrating the benefits of a bortezomib-based regimen in combination with PE for IgD MM with AKD.

Regression of renal amyloid deposits by VAD therapy plus autologous stem cell transplantation in a patient with primary AL amyloidosis.

We report a 58-year-old Japanese woman who presented with nephrotic syndrome. Steroid therapy and cyclosporine A administration were initiated, but hematological remission and renal response were not achieved. Renal biopsy revealed amyloid deposits in the mesangial region and the small arteries. Proteomic analysis based on laser microdissection and mass spectrometry showed that the amyloid deposits were composed of the constant region of the lambda light chain. She received vincristine, adriamycin, and dexamethasone therapy followed by high-dose melphalan and autologous stem cell transplantation, resulting in hematological complete remission and renal response with negative urinary Bence-Jones protein and proteinuria. Renal biopsy was performed four times during follow-up, demonstrating that amyloid deposits decreased gradually, while glomeruli showing global sclerosis increased from 3 to 62%. This case suggests that glomerular amyloid deposits can be cleared via tissue remodeling, if stem cells producing amyloid precursors are completely replaced by unrelated cells after stem cell transplantation.

Teste de cadeia leve livre - relação kappa/lambda para gamopatias monoclonais / Free light chain test - kappa / lambda ratio for monoclonal gammopathies

Contexto: As gamopatias monoclonais (paraproteinemias, disproteinemias ou imunoglobulinopatias), se caracterizam pela proliferação monoclonal de plasmócitos que produzem e secretam imunoglobulinas ou fragmentos de Imunoglobulina monoclonal (proteína monoclonal). A detecção e quantificação do componente monoclonal são necessárias para o diagnóstico. A eletroforese em gel de agarose e a imunofixação séricas e de urina são consideradas técnicas padrão. A pesquisa das cadeias leves livres (FLC) pode ser associada ao diagnóstico, posteriormente visando melhorar a taxa de detecção da doença. Pergunta: O teste de detecção de cadeias leves livres é sensível e específico para a detecção de cadeias leves livres, associado aos testes de eletroforese e imunofixação (imunoeletroforese) em soro e urina para diagnosticar pacientes com suspeita de gamopatia monoclonal, monitorar resposta à farmacoterapia de pacientes diagnosticados com gamopatia monoclonal, identificar recidiva precoce e avaliar prognóstico de pacientes? Evidências científicas: A evidência apresentada não incluiu todas as indicações solicitadas para a utilização do teste Freelite de dosagem de cadeias leves livres conforme descrito na pergunta de pesquisa. A evidência disponível é de baixa qualidade e apresenta limitação metodológica. Demonstra apenas o papel limitado do teste no diagnóstico de pacientes com suspeita de gamopatia monoclonal. Avaliação de Impacto Orçamentário: O demandante apresentou uma estimativa de impacto orçamentário para a utilização do teste de Freelite considerando apenas o diagnóstico de pacientes com suspeita de gamopatias monoclonais. Como o restante das indicações solicitadas, descritas na pergunta de pesquisa, não foram consideradas, nem se considerou os preços dos testes de dosagem de FLC concorrentes, a análise não apresentou valores confiáveis que possam suportar um pedido de incorporação da tecnologia no SUS. Decisão: Não incorporar o teste de cadeia leve livre - relação kappa/lambda para o diagnóstico de Gamopatias Monoclonais, no âmbito do Sistema Único de Saúde ­ SUS, dada pela Portaria SCTIE-MS nº 24 publicada no Diário Oficial da União (D.O.U.) nº 110, de 10 de junho de 2016.

Proteomics analysis of methylglyoxal-induced neurotoxic effects in SH-SY5Y cells.

Reactive carbonyl compounds contribute to aging, Alzheimer's disease (AD) and other neurodegenerative diseases. Among these compounds, methylglyoxal (MG) can yield advanced glycation end products (AGEs), which are crucial in AD pathogenesis. However, the molecular and biochemical mechanisms of MG neurotoxicity are not completely understood. In the present study, SH-SY5Y cells were treated with MG to induce cell death. 2-D Fluorescence Difference Gel Electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry were employed to determine the changes in protein levels in these cells compared with vehicle-treated cells. Proteomics analysis revealed that 49 proteins were differentially expressed in MG-treated SH-SY5Y cells, of which 16 were upregulated and 33 were downregulated. Among them, eight proteins were identified unambiguously. The significant changes in protein levels of actin, immunoglobulin lambda light chain and protein phosphatase 2 were noteworthy given their functional roles in AD pathogenesis. Taken together, our results suggest that multiple pathways are potentially involved in MG-induced neuron death.

Residual structures in the acid-unfolded states of Vlambda6 proteins affect amyloid fibrillation.

Many proteins form amyloid-like fibrils in vitro under partially or highly unfolding conditions. Recently, we showed that the residual structure in highly unfolded state is closely related to amyloid fibril formation in hen lysozyme. Thus, to better understand the role of the residual structure on amyloid fibril formation, we focused on AL amyloidosis, which results from the extracellular deposition of monoclonal immunoglobulin light-chain variable domains (V(L)s) as insoluble fibrils. We examined the relationship between the residual structure and amyloid fibril formation on three lambda6 recombinant V(L) (rVlambda6) proteins, wild type, Jto, and Wil. Although rVlambda6 proteins are highly unfolded in pH 2, (15)N NMR transverse relaxation experiments revealed nonrandom structures in regions, which include some hydrophobic residues and a single disulfide bond, indicating the existence of residual structure in rVlambda6 proteins. However, the residual structure of Wil was markedly disrupted compared with those of the other proteins, despite there being no significant differences in amino acid sequences. Fibrillation experiments revealed that Wil had a longer lag time for fibril formation than the others. When the single disulfide bond was reduced and alkylated, the residual structure was largely disrupted and fibril formation was delayed in all three rVlambda6 proteins. It was suggested that the residual structure in highly unfolded state has a crucial role in amyloid fibril formation in many proteins, even pathogenic ones.

The differential effects of dithiothreitol and 2-mercaptoethanol on the secretion of partially and completely assembled immunoglobulins suggest that thiol-mediated retention does not take place in or beyond the Golgi.

Dithiothreitol (DTT) blocks the endoplasmic reticulum (ER)-Golgi transport of newly synthesized immunoglobulin (Ig) molecules, whereas 2-mercaptoethanol (2ME) allows secretion of unpolymerized Igs otherwise retained intracellularly by disulphide interchange reactions. To understand this dichotomy, we have compared the effects of DTT and 2ME on the assembly, intracellular transport, and secretion of a panel of chimeric Igs that are either constitutively secreted or retained intracellularly. Our results demonstrate that DTT, but not 2ME, reduces some of the inter- and intrachain disulphide bonds and causes partial disassembly of H2L2 complexes and unfolding of individual chains in the ER. Upon DTT removal, heavy (H) and light (L) chains reform hapten-binding H2L2 molecules, which are later secreted. Reduction of the H2L2 interchain disulphide bonds can occur along the entire secretory pathway; however, in or beyond the Golgi this does not result in efficient H-L disassembly or unfolding. As a consequence, DTT does not block the exit from the Golgi. Moreover, unpolymerized Igs--normally retained in a pre-Golgi compartment--no longer require reducing agents to be secreted once they have reached the Golgi. Thus, little if any thiol-mediated retention seems to take place in or beyond the Golgi complex.