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Objective To evaluate the results of extracorporeal shockwave lithotripsy (ESWL) versus retrograde intrarenal surgery (RIRS) for the treatment of calcium oxalate nephrolithiasis, as well as the damaging effects on renal function, taking into account the dynamics of blood cystatin C and urine beta2-microglobulin.Material and Methods Of 94 patients with calcium oxalate nephrolithiasis aged 23–78 included in the study, 42 patients were classified as having undergone ESWL (group I) and 52 patients as having undergone RIRS (group II). Group II patients were then stratified into subset 2A (n = 32) as having undergone RIRS through rigid ureteroscope and subset 2B (n = 20) as having undergone RIRS through flexible ureteroscope. We performed plain urography and nephrosonography at 24–48 hours postoperatively and unenhanced computed tomography 4–6 weeks after surgery. We measured concentrations of serum cystatin C and urinary beta2-microglobulin as a marker for kidney damage. In group I, samples of peripheral blood andurine were taken before and after the first, third sessions and 30 days after the last ESWL session. In group II, samples were analyzed before surgery, on the first and 30th postoperative days.Results The average size of calculi in the group with RIRS was 16.91 ± 2.79 mm, in the group with ESWL 12.31 ± 2.27 mm. The need for reoperation after RIRS was 19.2%, which was lower than after ESWL. Stone-free effect (no stones, or residual stones less than 3 mm) was observed in 95% of cases in patients with RIRS, and in 78% with ESWL. Group I patients demonstrated an increase in the blood leukocytes total number more often than subsets 2A (rigid RIRS) and 2B (flexible RIRS) patients. Leukocyturia was also a more common complication in group I. In the RIRS group, there was no statistically significant change in the level of blood cystatin C and urine beta2-microglobulin, on the contrary, a moderate increase in the endogenous marker of cystatin C was noted after one ESWL session. The increase in urine beta2-microglobulin levels in patients after the first and third ESWL sessions was significantly higher than after RIRS.Conclusion Flexible RIRS may be suggested as the preferred procedure for patients requiring additional protection of renal function in the treatment of renal stones less than 20 mm. ESWL of stones less than 20 mm can be used as an alternative treatment, since it is characterized by a rather long period of stone eradication from the urinary tract, a high frequency of residual calculi after the procedure, and also has a damaging effect on the renal tissue.

Total or partial loss of HLA class I antigens reduce the recognition of specific tumor peptides by cytotoxic T lymphocytes favoring cancer immune escape during natural tumor evolution. These alterations can be caused by genomic defects, such as loss of heterozygosity at chromosomes 6 and 15 (LOH-6 and LOH-15), where HLA class I genes are located. There is growing evidence indicating that LOH in HLA contributes to the immune selection of HLA loss variants and influences the resistance to immunotherapy. Nevertheless, the incidence and the mechanism of this chromosomal aberration involving HLA genes has not been systematically assessed in different types of tumors and often remains underestimated. Here, we used SNP arrays to investigate the incidence and patterns of LOH-6 and LOH-15 in a number of human cancer cell lines and tissues of different histological types. We observed that LOH in HLA is a common event in cancer samples with a prevalence of a copy neutral type of LOH (CN-LOH) that affects entire chromosome 6 or 15 and involves chromosomal duplications. LOH-6 was observed more often and was associated with homozygous HLA genotype and partial HLA loss of expression. We also discuss the immunologic and clinical implications of LOH in HLA on tumor clonal expansion and association with the cancer recurrence after treatment., ISCIII Research Institute co-financed by the European Union and Ministerio de Ciencia e Innovación (FEDER-Fondo Europeo de Desarrollo Regional and National Plan 2020, MICIN) (PI17/00197, PI18/00826, and PID2020-115087GB-100), Junta de Andalucía in Spain (Group CTS-143 and B-CTS-410-UGR-20), Consejería de Salud, Junta de Andalucía through the contract “Nicolás Monardes” [C-0013-2018]

Nguyen Huu Dung,1 Nguyen Trung Kien,2 Nguyen Thi Thu Hai,1 Phan The Cuong,1 Nguyen Thi Thu Huong,3 Dao Bui Quy Quyen,4 Nguyen Minh Tuan,4 Do Manh Ha,2 Truong Quy Kien,2 Nguyen Thi Thuy Dung,2 Pham Quoc Toan,2 Hoang Trung Vinh,2 Tomoko Usui,5 Le Viet Thang21Bach Mai Hospital, Ha Noi, Vietnam; 2Military Hospital 103, Ha Noi, Vietnam; 3Ha Noi Kidney Hospital, Ha Noi, Vietnam; 4Cho Ray Hospital, Ho Chi Minh, Vietnam; 5University of Tokyo Hospital, Tokyo, JapanPurpose: Beta2-microglobulin (β2-M) is recognized as a surrogate marker relating to the mechanisms of dialysis-associated amyloidosis. Few studies have evaluated the association of serum β2-M with clinical outcome in hemodialysis patients using high-flux type. However, study on patients using low-flux dialyzer reuse has not been done yet.Patients and methods: Using serum β2-M level on predicting long-term mortality of hemodialysis patients was examined in 326 prevalent hemodialysis patients (45.59±14.46 years, hemodialysis duration of 47.5 (26–79) months, 186 males and 140 females). The patients were divided into 3 groups with equal number of patients, according to their serum β2-M levels: group A (n=109, serum β2-M concentration ≤55.7 mg/L), group B (n=109, serum β2-M level from 55.8 mg/L to 75.4 mg/L) and group C (n=108, serum β2-M concentration >75.4 mg/L).Results: During the follow-up period of 5 years, there were 75 all-cause deaths (23.0%). Kaplan–Meier analysis revealed that all-cause mortality in the higher β2-M group was significantly higher compared to that in the lower β2-M groups (p

Beta2-microglobulin (B2M) a key component of major histocompatibility complex class I molecules, which aid cytotoxic T-lymphocyte (CTL) immune response. However, the majority of studies of B2M have focused only on amyloid fibrils in pathogenesis to the neglect of its role of antimicrobial activity. Indeed, B2M also plays an important role in innate defense and does not only function as an adjuvant for CTL response. A previous study discovered that human aggregated B2M binds the surface protein structure in Streptococci, and a similar study revealed that sB2M-9, derived from native B2M, functions as an antibacterial chemokine that binds Staphylococcus aureus. An investigation of sB2M-9 exhibiting an early lymphocyte recruitment in the human respiratory epithelium with bacterial challenge may uncover previously unrecognized aspects of B2M in the body’s innate defense against Mycobactrium tuberculosis. B2M possesses antimicrobial activity that operates primarily under pH-dependent acidic conditions at which B2M and fragmented B2M may become a nucleus seed that triggers self-aggregation into distinct states, such as oligomers and amyloid fibrils. Modified B2M can act as an antimicrobial peptide (AMP) against a wide range of microbes. Specifically, these AMPs disrupt microbe membranes, a feature similar to that of amyloid fibril mediated cytotoxicity toward eukaryotes. This study investigated two similar but nonidentical effects of B2M: the physiological role of B2M, in which it potentially acts against microbes in innate defense and the role of B2M in amyloid fibrils, in which it disrupts the membrane of pathological cells. Moreover, we explored the pH-governing antibacterial activity of B2M and acidic pH mediated B2M amyloid fibrils underlying such cytotoxicity.

Background. Compared to high-flux dialysis membranes, novel medium cut-off (MCO) membranes show greater permeability for larger middle molecules. Methods. In two prospective, open-label, controlled, randomized, crossover pilot studies, 39 prevalent hemodialysis (HD) patients were studied in four dialysis treatments as follows: study 1, three MCO prototype dialyzers (AA, BB and CC with increasing permeability) and one high-flux dialyzer in HD; and study 2, two MCO prototype dialyzers (AA and BB) in HD and high-flux dialyzers in HD and hemodiafiltration (HDF). Primary outcome was lambda free light chain (λFLC) overall clearance. Secondary outcomes included overall clearances and pre-to-post-reduction ratios of middle and small molecules, and safety of MCO HD treatments. Results. MCO HD provided greater λFLC overall clearance [least square mean (standard error)] as follows: study 1: MCO AA 8.5 (0.54), MCO BB 11.3 (0.51), MCO CC 15.0 (0.53) versus high-flux HD 3.6 (0.51) mL/min; study 2: MCO AA 10.0 (0.58), MCO BB 12.5 (0.57) versus high-flux HD 4.4 (0.57) and HDF 6.2 (0.58) mL/min. Differences between MCO and high-flux dialyzers were consistently significant in mixed model analysis (each P < 0.001). Reduction ratios of λFLC were greater for MCO. Clearances of α1-microglobulin, complement factor D, kappa FLC (κFLC) and myoglobin were generally greater with MCO than with high-flux HD and similar to or greater than clearances with HDF. Albumin loss was moderate with MCO, but greater than with high-flux HD and HDF. Conclusions. MCO HD removes a wide range of middle molecules more effectively than high-flux HD and even exceeds the performance of high-volume HDF for large solutes, particularly λFLC.

DNA mismatch repair (MMR)-deficient cancers accumulate high numbers of coding microsatellite mutations, which lead to the generation of highly immunogenic frameshift peptide (FSP) neoantigens. MMR-deficient cells can grow out to clinically manifest cancers either if they evade immune cell attack or if local T-cells get exhausted. Therefore, a subset of MSI cancer patients responds particularly well to treatment with immune checkpoint inhibitors. We analyzed whether immune evasion in MMR-deficient cancer mediated by loss of HLA class I or II antigens is related to local immune cell activation status. Microsatellites located in Beta2-microglobulin (B2M) and the HLA class II-regulatory genes RFX5 and CIITA were analyzed for mutations in MMR-deficient colorectal cancers (n = 53). The results were related to CD3-positive and PDCD1 (PD-1)-positive T-cell infiltration. PDCD1 (PD-1)-positive T-cell counts were significantly higher in B2M-mutant compared to B2M-wild type tumors (median: 22.2 cells per 0.25 mm2 vs. 2.0 cells per 0.25 mm2, Wilcoxon test p = 0.002). Increasing PDCD1 (PD-1)-positive T-cell infiltration was significantly related to an increased likelihood of B2M mutations (OR = 1.81). HLA class II antigen expression status was significantly associated with enhanced overall T-cell infiltration, but not related to PDCD1 (PD-1)-positive T-cells. These results suggest that immune evasion mediated by B2M mutation-induced loss of HLA class I antigen expression predominantly occurs in an environment of activated PDCD1 (PD-1)-positive T cell infiltration. If B2M mutations interfere with anti-PDCD1 (PD-1)/CD274 (PD-L1) therapy success, we predict that resistance towards anti-PDCD1 (PD-1) therapy may – counterintuitively – be particularly common in patients with MMR-deficient cancers that show high PDCD1 (PD-1)-positive T cell infiltration.

Real-time reverse transcription polymerase chain reaction (RT-PCR) is regarded as one of the most useful and powerful tools for characterizing hematopoietic stem cells (HSCs), because samples of extremely small cell numbers can be analyzed. The expression levels determined by RT-PCR are based on relative quantification; therefore, the selection of an appropriate reference gene with a relatively stable expression level under most conditions is crucial. Here, we determined that beta2-microglobulin (B2m) is an appropriate reference gene for analyzing mouse HSCs by a novel method using single-cell RT-PCR. Clonally sorted HSCs were subjected to RT reactions with exogenous RNA fragments and then to real-time PCR. Next, the relative gene expression levels of 4 well-known housekeeping genes were quantified in each single cell sample based on the threshold cycle of exogenous RNA. The analysis revealed that B2m expression was reproducibly detected in almost all HSCs and that B2m had the most stable expression level among the compared genes, even after the cells had been cultured under various conditions. Thus, our results indicate that B2m can reliably be used as a reference gene for the relative quantification of expression levels in HSCs across various conditions. Furthermore, our work proposes a novel method for the selection of appropriate reference genes.

The oriented immobilization of proteins, key for the development of novel responsive biomaterials, relies on the availability of effective probes. These are generally provided by standard approaches based on in vivo maturation and in vitro selection of antibodies and/or aptamers. These techniques can suffer technical problems when a non-immunogenic epitope needs to be targeted. Here we propose a strategy to circumvent this issue by in silico design. In our method molecular binders, in the form of cyclic peptides, are computationally evolved by stochastically exploring their sequence and structure space to identify high-affinity peptides for a chosen epitope of a target globular protein: here a solvent-exposed site of β2-microglobulin (β2m). Designed sequences were screened by explicit solvent molecular dynamics simulations (MD) followed by experimental validation. Five candidates gave dose-response surface plasmon resonance signals with dissociation constants in the micromolar range. One of them was further analyzed by means of isothermal titration calorimetry, nuclear magnetic resonance, and 250 ns of MD. Atomic-force microscopy imaging showed that this peptide is able to immobilize β2m on a gold surface. In short, we have shown by a variety of experimental techniques that it is possible to capture a protein through an epitope of choice by computational design.

More potent antitumor activity is desired in Interferon (IFN)-treated cancer patients. This could be achieved by combining IFN alpha and IFN gamma. The aim of this work was to characterize the pharmacokinetics and pharmacodynamics of a novel formulation containing a co-formulated combination of IFNs alpha-2b and gamma (CIGB-128-A). A group of nine healthy male subjects received intramuscularly 24.5 × 106 IU of CIGB-128-A. IFN concentrations were evaluated for 48 h. Serum neopterin, beta2-microglobulin (β2M) and 2′–5′ oligoadenylate synthetase (2′–5′ OAS), classical IFN-inducible serum markers, were measured during 192 h by enzyme immunoassay and body temperature was used as pharmacodynamic variable as well. Concerning pharmacokinetics, serum IFNs’ profiles were better fitted to a mono-compartmental model with consecutive zero order and first order absorption, one bioavailability value. No interferences by simultaneous administered IFNs were observed in their typical similar systemic profiles. Neopterin and β2M time profiles showed a delay that was efficiently linked to pharmacokinetics by means of a zero order absorption rate constant. Neopterin level was nine-fold higher than initial values, 48 h post-administration, an increment not described before. At this time, mean serum β2M peaked around the double from baseline. Serum concentrations of the enzyme 2′–5′ OAS was still elevated on the 8 day post-injection. The formulation was well tolerated. Most frequent adverse reactions were fever, headache, arthralgia and lymphopenia, mostly mild. The administration of co-formulated IFN alpha-2b and IFN gamma likely provides improved pharmacodynamic properties that may be beneficial to treat several malignancies. Cuban Public Registry of Clinical Trials RPCEC00000118 , May 24, 2011.

Background: High-level microsatellite instability (MSI-H) has been reported as a prognostic marker in colon cancer. We here analysed the prognostic significance of MSI and mutations of the Beta2-Microglobulin (B2M) gene, which occur in about 30% of MSI-H colon cancer, in the cohort of the prospective FOGT-4 (Forschungsruppe Onkologie Gastrointestinale Tumoren, FOGT) trial. Methods: Microsatellite instability status was determined using standard protocols (NCI/ICG-HNPCC panel and CAT25) in 223 colon cancer lesions. Beta2-Microglobulin mutation status was evaluated by exon-wise sequencing in all MSI-H lesions. Results: Patients with MSI-H (n=34) colon cancer presented with a significantly lower risk of relapse after 12 months of follow-up compared with MSS (n=189) colon cancer patients (5 year time to relapse: MSI-H 0.82 vs MSS 0.66, P=0.03). No significant difference in overall survival was detected. Beta2-Microglobulin mutations were identified in 10 (29.4%) out of 34 MSI-H colon cancers and were associated with a complete absence of disease relapse or tumour-related death events (P=0.09). Conclusion: The risk of late disease relapse was significantly lower in patients with MSI-H compared with MSS colon cancer. Moreover, B2M mutations may contribute to the favourable outcome of MSI-H colon cancer patients and should therefore be evaluated as a potential prognostic marker in future clinical trials.

Mismatch repair (MMR)-deficient cancers accumulate multiple insertion/deletion mutations at coding microsatellites (cMS), which give rise to frameshift peptide neoantigens. The high mutational neoantigen load of MMR-deficient cancers is reflected by pronounced anti-tumoral immune responses of the host and high responsiveness towards immune checkpoint blockade. However, immune evasion mechanisms can interfere with the immune response against MMR-deficient tumors. We here performed a comprehensive analysis of immune evasion in MMR-deficient colorectal cancers, focusing on HLA class I-mediated antigen presentation. 72% of MMR-deficient colorectal cancers of the DFCI database harbored alterations affecting genes involved in HLA class I-mediated antigen presentation, and 54% of these mutations were predicted to abrogate function. Mutations affecting the HLA class I transactivator NLRC5 were observed as a potential new immune evasion mechanism in 26% (6% abrogating) of the analyzed tumors. NLRC5 mutations in MMR-deficient cancers were associated with decreased levels of HLA class I antigen expression. In summary, the majority of MMR-deficient cancers display mutations interfering with HLA class I antigen presentation that reflect active immune surveillance and immunoselection during tumor development. Clinical studies focusing on immune checkpoint blockade in MSI cancer should account for the broad variety of immune evasion mechanisms as potential biomarkers of therapy success.

As the lymphotropism of hepatitis C virus (HCV) has already been ascertained, and in the light of the fact that the immune defense system is an organized network composed of functionally interrelated tissues, this study was carried out to verify the possible involvement of spleen in HCV-related chronic hepatitis. In this cross-sectional study we measured spleen longitudinal diameter by ultrasound, beta2-microglobulin serum levels and splenic artery resistivity index (SARI) by Doppler in 51 patients treated with standard combined (Peg-Interferon plus Ribavirin) antiviral therapy. Thirty-three patients (17 females) completed the regimen and were compared to 31 controls (16 females). The mean basal values of spleen longitudinal diameter were higher in patients with chronic hepatitis than in controls, i.e., 116 mm (9.4) versus 102.7 mm (9.3), P = 0.0001. In the same patients a significant trend towards increased spleen longitudinal diameter was found after antiviral therapy, independently of the stage of HCV-related chronic hepatitis. The median values of the beta2-microglobulin concentrations were not significantly higher in the patients with HCV-related chronic hepatitis compared to controls, i.e., 1.3 (0.5 – 2.6) versus 1 (0.6 – 1.4), P = 0.16, although during the course of therapy they were significantly increased. SARI values of HCV-related chronic hepatitis patients were different from those of controls, but were unvaried compared to values at the end of treatment. Neither spleen measurements nor serum beta2-microglobulin levels were able to predict therapeutic response to antiviral therapy. A stimulation/ expansion of lymphoid tissue was found in patients with HCV-related chronic hepatitis. Such evidence raises the question whether physicians should continue to prescribe antiviral therapy in non-responders and supports the use of a new scheme (SLD plus β2-MG) to diagnose this ongoing, apparently reversible but nevertheless dangerous immunologic process.

To analyze the in vivo biological effect of anti-interferon beta (IFN-beta) neutralizing antibodies (NABs), blood concentrations of neopterin, beta2microglobulin (Beta2-MG), mRNA-dependent myxovirusresistant protein A (MxA) and dsRNA-dependent protein kinase (PKR) were measured before (predose) and 24 hours after (postdose) IFN-beta administration in 49 patients with multiple sclerosis (MS) with (n = 25) and without (n = 24) NABs. The results indicated that predose levels of MxA-mRNA and PKR-mRNA were highly variable [coefficient of variation (CV) > 100%] among patients. A lower inter-individual variability was observed for pre-dose levels of Beta2-MG and neopterin (CVs of 29% and 44%, respectively). Significantly lower pre- and post-dose blood levels of IFN induced markers, except for postdose PKR-mRNA (p = 0.09), were seen in NAB+ compared with NAB-patients and between patients with high (> 200 t1/10) and low (£ 200 t1/10) NAB titers. A significant inverse correlation between NAB titer and pre-dose levels of the above IFN-induced markers was found. In summary, our findings confirm that NABs affect absolute concentrations of IFN-beta induced markers and suggest that such an effect occurs in a titer-dependent manner

Objectives: Men who have sex with men are at risk of tenofovir nephrotoxicity due to its wide use in both treatment and prophylaxis for human immunodeficiency virus infection, but little is known about the urinary biomarkers of early renal dysfunction in this population. This study aims to identify useful biomarkers of early renal dysfunction among human immunodeficiency virus-infected men who have sex with men exposed to tenofovir.Methods: In a cross-sectional study urinary alpha1-microglobulin, beta2-microglobulin, N-acetyl-B-n-glucosaminidase and albumin were measured and expressed as the ratio-to-creatinine in 239 human immunodeficiency virus-infected men who have sex with men who were treatment naïve or receiving antiretroviral therapy with tenofovir-containing or non-tenofovir-containing regimens. Additionally, 56 patients in the non-antiretroviral therapy group started a tenofovir-containing regimen and were assessed after 3 and 6 months on antiretroviral therapy.Results: Both the frequency of alpha1-microglobulin proteinuria (alpha1-microglobulin-creatinine ratio >25.8 mg/g) and the median urinary alpha1-microglobulin-creatinine ratio were higher in the tenofovir disoproxil fumarate group than the other two groups (all p< 0.05). A higher frequency of beta2-microglobulin proteinuria (beta2-microglobulin-creatinine ratio >0.68 mg/g) was also observed in the tenofovir group (28.9%) compared to the non-tenofovir group (13.6%, p= 0.024). There were no significant differences between groups for N-acetyl-β-n-glucosaminidase and albumin. In the longitudinal study, the median urinary alphat-microglobulin-creatinine ratio after 3 and 6 months on tenofovir-containing therapy (16.8 and 17.3 mg/g) was higher than baseline (12.3 mg/g, p= 0.023 and 0.011, respectively), while no statistically important changes were observed in urinary beta2-microglobulin-creatinine ratio or in the other biomarkers after 3 and 6 months on antiretroviral therapy (all p> 0.05).Conclusion: Urinary alphat-microglobulin seems to be a more sensitive and stable indicator of tubular dysfunction than urinary beta2-microglobulin for assessing tenofovir-related nephrotoxicity and can be significantly altered after tenofovir exposure.

Background The main short-term advantages of haemodiafiltration (HDF) are supposedly better removal of Beta2-microglobulin (ß2-m) and phosphate, and better haemodynamic stability. The main disadvantage is higher costs. The aim of the study was to compare the clinical and biological parameters associated with HDF and high-flux haemodialysis (HD), using a cross-over design, while maintaining the same dialysis parameters. Methods All patients on a 3 × 4 hours schedule were observed during 3 identical 6-months periods: HDF1 – HD – HDF2. The mean values for the 2 last months of each period were compared. Results A total of 51 patients (76 % males, 45 % diabetic) with a mean age of 74 ± 15 years, and who had been on dialysis for 49 ± 60 months were included. The mean blood flow (329 ± 27 ml/min), dialysate flow (500 ml/min), and convection volumes (21.6 ± 3.2 L) were recorded. Patient medications were not changed. Predialysis blood pressure, phosphataemia, calcaemia, iPTH, Kt/V, nPNA and intradialytic events were similar throughout the 3 periods. Only serum albumin (34. 4 ± 3.6, 35.9 ± 3.4, 34.1 ± 4 g/L, p

Background Hyperglycemia appears to play a significant role on the inflammatory cytokines production. Beta2-microglobulin (beta2M) is accumulated in the circulation of dialysis patients. We studied the relationship between glycemic control defined by glucose serum concentrations and insulin resistance, beta2M and markers of inflammation in patients on renal replacement therapies with or/and without diabetes mellitus. Methods We enrolled 96 dialyzed patients, 62 males and 34 females. The treatment modalities which were applied were : regular hemodialysis (HD, n = 34), predilution hemodiafiltration (HDF, n = 42) and peritoneal dialysis (PD, n = 20). Dialysis adequacy was defined by Kt/V for urea.Beta2M and insulin serum concentrations were measured by radioimmunoassays. hsCRP and TNF-α serum concentrations were measured by ELISA. Insulin resistance was calculated using the homeostasis model assessment of insulin resistance (HOMA-IR).We examined the association of elevated serum glucose with inflammatory factors and we built a multivariable model to investigate if glucose could be a potential determinant of beta2M serum levels. Results Serum glucose was positively correlated with beta2M and TNF-α (r = 0.320, p = 0.002 and r = 0.215, p = 0.03 respectively).We observed significant association between the patients with higher serum glucose concentrations and the patients with greater beta2Μ concentrations (x2 = 4.44, p = 0.03). Multivariable model showed that glucose acts as a significant independent determinant of beta2M adjusting for age, gender, dialysis modality and metabolic acidosis status. Conclusions The elevated glucose concentrations were positively associated with both, greater beta2M serum concentrations and up-regulated inflammatory procedure in dialysis patients with or/and without diabetes mellitus.

Das fibrillen-bildende beta2-Mikroglobulin (b2M) und das CRF1-Mimetikum mit verzweigter Rückgratstruktur können als „schwierige“ Proteine betrachtet werden, deren Darstellung sich eignet, gegenwärtige Möglichkeiten und Grenzen der Proteinsynthese zu ermitteln. Die Proteine sollen zu spektroskopischen Untersuchungen von Proteinfaltung bzw. Ligand-Rezeptor- Wechselwirkungen eingesetzt werden. Versuche zur Chemosynthese von b2M über drei Segmente führten per NCL zwar zu linearen Produkten mit korrekter Primärstruktur, aber wiederholt wurden zwei, mittels HPLC trennbare Proteine erhalten, deren enzymatische Spaltung zu identischen Fragmenten führte. Eine Isomerisierung (wie z.B. Epimerisierung) als Ursache für die Bildung der zwei Produkte konnte ausgeschlossen werden. Mittels CD- und FTIR-Spektroskopie wurden für beide Produkte beta- Faltblatt-Strukturen ermittelt, die sich sowohl untereinander als auch vom rekombinanten Protein unterschieden. Die „fehlgefalteten“ Syntheseprodukte konnten nicht entfaltet und anschließend in die „korrekte“ Struktur des rekombinanten b2M überführt werden. Es ist denkbar, dass die beobachtete „Fehlfaltung“, deren Ursache nicht geklärt werden konnte, für vom b2M ausgelöste Amyloidosen verantwortlich ist. Das CRF1-Modell, das aus drei zyklischen Peptiden und einem Protein mit Disulfidbrücken besteht, welche auf einem linearen Peptid-Templat verankert sind, wurde durch ein zyklisches Templat zur strukturellen Einschränkung modifiziert. Durch das zyklische Templat ergaben sich keine Syntheseprobleme, aber interessanterweise führte die Zyklisierung des Templats zu einer signifikant höheren Affinität für den Agonisten Urocortin-I im funktionellen Assay. Darüber hinaus wurde gezeigt, dass ein zyklisches Rezeptor-Loop-Peptid mittels EPL im mg-Maßstab erhalten werden kann, was künftig die Synthese isotopenmarkierter Analoga für Struktur-Untersuchungen ermöglicht. The fibril forming beta2-microglobulin (b2m) and the CRF1 mimic with branched peptide backbone could be considered as “difficult” proteins, whose synthesis is suited for determining present possibilities and limits of protein synthesis. The proteins shall be used for spectroscopic analysis of protein misfolding or ligand-receptor-interaction, respectively. Efforts of the chemosynthesis of b2m over three segments may lead via NCL to linear products with correct primary structure, but two, via HPLC isolatable proteins were repetitively susbstained, whose enzymatic digest lead to identical fragments. An isomerization (such as e. g. epimerization) as reason for the formation of the two products could be excluded. By means of CD and FTIR spectroscopy for both products beta-sheet structure were determined, which differ among themselves as well as from the recombinant protein. The “misfolded” synthetic product could not be unfolded und subsequently converted into the “correct” structure of the recombinant b2m. It is possible that the observed “misfolding”, whose cause could not be clarified, is reasonable for the amyloidosis induced by b2m. The CRF1 model that consists of three cyclic peptides and one protein with disulfid bridges coupled to a linear peptide template, was modified for structural constraints by a cyclic template. In consequence of the cyclic template no synthetic problems aroused, although the cyclisation of the template leads interestingly to a significant higher affinity for the antagonist urocortin-I in the functional assay. Furthermore, it was shown that a cyclic receptor loop peptide could be received via EPL in mg scale, what in future enables the synthesis of isotopically labeled analogs for structure investigations.

Dialysis-related amyloidosis (DRA) is one of the most important complications in long-term dialysis patients. Pulmonary involvement in patients with DRA has been rarely described, and lung radiographic findings have not yet been reported. The most common chronic lung disease process in chronic dialysis patients is interstitial fibrosis. This is the first case report of DRA presenting in the lung in a manner resembling interstitial pneumonia. This case study suggests that interstitial pneumonia as a result of DRA should be considered when dyspnoea and reticular opacity of the lung are observed in patients undergoing long-term dialysis.

Characterization of beta2-microglobulin in a primitive fish, the Siberian sturgeon (Acipenser baeri)

Le syndrome du canal carpien (SCC) regroupe l'ensemble des signes et symptômes liés à la compression du nerf médian dans le canal carpien. Cette manifestation de l'amylose à béta2-microglobuline est une complication fréquente de l'hémodialyse au long cours. L'objectif de ce travail est d'analyser les caractéristiques du SCC et de déterminer les facteurs liés à sa survenue chez les hémodialysés chroniques.Nous rapportons une étude transversale monocentrique, menée au 3ème trimestre de l'année 2009, portant sur les patients adultes hémodialysés chroniques au service de Néphrologie-Hémodialyse au CHU de Fès. 59 patients ont accepté de participer à l'étude. Leurs âge moyen est de 48 ± 15 ans avec un sex-ratio de 0,9. Ils bénéficient tous de l'hémodialyse intermittente à raison de 10 à 12 heures par semaine, par une membrane de dialyse en polysulfone à basse perméabilité. La durée moyenne en hémodialyse est de 83 ± 6,5 mois. La prévalence du SCC dans notre centre est de 30,5%. L'électromyogramme (EMG) a confirmé la suspicion clinique du SCC chez 11 patients et a diagnostiqué un SCC chez 8 patients asymptomatiques. La comparaison statistique entre les deux groupes de patients avec et sans SCC a démontré que la survenue de ce syndrome est liée à: l'âge actuel, l'âge avancé à la mise en hémodialyse, le sexe féminin, l'excès pondéral, et l'abord vasculaire. Le SCC est une complication fréquente de l'hémodialyse chronique. L'amélioration de la qualité de dialyse permettrait de réduire le risque de survenue du SCC.Pan African Medical Journal 2013; 14:19

The amyloid pathology associated with long-term haemodialysis is due to the deposition of ß2-microglobulin, the non-polymorphic light chain of class I major histocompatibility complex, that accumulates at bone joints into amyloid fibrils. Several lines of evidence show the relevance of the tryptophan residue at position 60 for the fibrillogenic transition of the protein. A comparative 15N NMR relaxation analysis is presented for wild-type human ß2-microglobulin and W60G ß2- microglobulin, i.e. the mutant with a glycyne replacing the natural tryptophan residue at position 60. The experimental data, collected at 11.4 T and 310 K, were analyzed by means of the reduced spectral density approach. Molecular dynamics (MD) simulations and corresponding thermodynamic integration, together with hydrodynamic calculations were performed to support data interpretation. The analysis results for the mutant protein are consistent with a reduced aggregation with respect to the wild-type counterpart, as a consequence of an increased conformational rigidity probed by either NMR relaxation and MD simulations. Although dynamics in solution is other than fibrillar competence, the assessed properties of the mutant protein can be related with its reduced ability of forming fibrils when seeded in 20% trifluoroethanol. Copyright © 2013 John Wiley & Sons, Ltd.

The amyloid pathology associated with long-term haemodialysis is due to the deposition of (2)-microglobulin, the non-polymorphic light chain of class I major histocompatibility complex, that accumulates at bone joints into amyloid fibrils. Several lines of evidence show the relevance of the tryptophan residue at position 60 for the fibrillogenic transition of the protein. A comparative N-15 NMR relaxation analysis is presented for wild-type human (2)-microglobulin and W60G (2)-microglobulin, i.e. the mutant with a glycyne replacing the natural tryptophan residue at position 60. The experimental data, collected at 11.4T and 310K, were analyzed by means of the reduced spectral density approach. Molecular dynamics (MD) simulations and corresponding thermodynamic integration, together with hydrodynamic calculations were performed to support data interpretation. The analysis results for the mutant protein are consistent with a reduced aggregation with respect to the wild-type counterpart, as a consequence of an increased conformational rigidity probed by either NMR relaxation and MD simulations. Although dynamics in solution is other than fibrillar competence, the assessed properties of the mutant protein can be related with its reduced ability of forming fibrils when seeded in 20% trifluoroethanol. Copyright (c) 2013 John Wiley & Sons, Ltd. MIURMinistero dell' Istruzione, dell' Universita e della Ricerca (MIUR) [RBRN07BMCT, 20083ERXWS]; EUEuropean Union (EU) [LSHM-CT-2005-037525] This work was supported by MIUR (grants FIRB No. RBRN07BMCT, PRIN No. 20083ERXWS), and by EU (EURAMY project LSHM-CT-2005-037525). Dr Rotter M. and D. Makek A. are acknowledged for the help with calculations and assistance. DG thanks Dr Alexandros S. Papadopoulos for the helpful discussion on statistical aspects of data analysis.

Under particular conditions proteins spontaneously aggregate into well-ordered structures called amyloid fibrils. The full elucidation of the fibrillation process requires the identification by biophysical techniques of the conformational and oligomeric species populated during the aggregation mechanism.

Human immunodeficiency virus type 1 (HIV-1) invades the central nervous system (CNS) early in the course of infection and either directly or through opportunistic infections causes a spectrum of neurological complications. The most severe manifestation of HIV-1 CNS infection is AIDS Dementia Complex (ADC), which occurs in approximately 20% of untreated patients with AIDS. ADC is considered the result of a complex interplay between immune activation effects and viral replication in the brain, which ultimately leads to neuronal injury and death. Reliable markers to diagnose HIV-1 associated CNS injury, track disease progression, and identify patients at risk of developing ADC are lacking. Such markers would also be beneficial in evaluating the efficacy of antiretroviral treatment (ART) in the CNS, as well as to provide insights into the pathogenesis of HIV-1 CNS infection.HIV-1 elicits intrathecal cell-mediated and humoral immune activation. We found that ART effectively decreased the cerebrospinal fluid (CSF) concentrations of neopterin and beta2-microglobulin, but had little effect on the elevated IgG index. However, almost half of the patients still had slightly elevated levels of neopterin after 2 years of follow-up. Phylogenetic analyses have identified 3 distinct HIV-1 genetic groups. Group M, which is responsible for most of the global HIV-1 epidemic is further subdivided into subtypes and circulating recombinant forms (CRFs). Most of the current knowledge of HIV-1 CNS infection is based on studies of subtype B, which is predominant in the western world. However, subtypes other than subtype B are responsible for most of the epidemic outside the western world, and HIV-1 infections due to subtypes other than B are rapidly increasing across Europe. Markers of HIV-1 CNS infection such as HIV-1 RNA, neopterin, and white blood cell (WBC) count in CSF were measured and compared in patients infected with different HIV-1 subtypes. We did not find any significant subtype-specific differences in the neuromarkers evaluated in this study. Thus, subtypes do not appear to influence neuropathogenesis.Although there is no evidence of productive infection of neurons the end-result of HIV-1 CNS infection is neuronal damage and loss. We investigated the potential of CSF neurofilament (NFL), a sensitive indicator of axonal injury, as a marker of HIV-1 associated neurodegeneration. CSF NFL concentrations were higher in patients with ADC than in neuroasymptomatic patients, or patients with primary HIV-1 infection. Patients with severe ADC had higher CSF NFL levels compared to those with milder disease. CSF NFL declined with ART to the limit of detection in parallel with virological response and neurological improvement in patients suffering from ADC.Neurocognitive impairment remains a major concern in HIV-1 infection despite the success of ART. Studies on the pathogenesis, epidemiology, and the effects of ART on HIV-1 CNS infection are important to improve patient management.

Convective and adsorptive removal of β2-microglobulin during predilutional and postdilutional hemofiltration. Background Beta 2 -microglobulin (β2-m) removal in patients with end-stage renal disease (ESRD) is maximal with convective techniques, such as hemofiltration (HF) or hemodiafiltration (HDF). Although the infusion mode of the replacement solution (predilution or postdilution) is expected to influence the efficiency of HF, experimental data in this respect are scanty. We therefore investigated the impact of the fluid reinfusion mode on the efficiency of HF in 11 ESRD patients who underwent both treatments. Methods The dialyzer (AK 200 ULTRA) was equipped with a 3-layer polyamide membrane (Poliflux 21 S, surface 2.1m 2 ) and blood flow was kept between 300 and 400mL/min. β2-m concentrations were measured in plasma water and ultrafiltrate at appropriate times during a 240-minute treatment. The following dialytic parameters were calculated: total amount of β2-m removed (A tot ), β2-m removed by convection (A con ) and by adsorption (A ads ), percent reduction in β2-m plasma water concentration (% Cpw in ), total plasma water clearance (CLpw tot ), convective plasma water clearance (CLpw con ), adsorptive plasma water clearance (CLpw ads ), and sieving coefficient (SC). Results CLpw tot , CLpw ads , and% Cpw in were similar in pre- and postdilutional conditions, whereas CLpw con and SC were higher and CLpw ads was lower in postdilution than in predilution HF. Since a significant inverse correlation was found between A ads and SC, predilution probably determines greater protein fouling than postdilution. Conclusion The 2 techniques appear to be equivalent in terms of total β2-m removal, although this final result is obtained by different contributions of convective and adsorptive elimination.

Interferon-beta 1b (IFN-beta 1b) führt in der Therapie der Multiplen Sklerose (MS) zu einer signifikanten Reduktion von Frequenz und Schweregrad der Schübe. Bisher ist die Wirkweise von IFN-beta 1b nicht vollständig geklärt, Zytokinen wird eine Schlüsselrolle zugesprochen. In der vorliegenden Studie wurden die folgenden 10 (pro- und antiinflammatorischen) Zytokine mittels ELISA im Serum von 23 MS-Patienten über einen Zeitraum von drei Monaten zu sechs verschiedenen Zeitpunkten bestimmt: TNF-alpha, sTNF-R, IL-6, sIL-6R, IL-1beta, IL-1ra, sIL-2R, IL-10, Neopterin, Beta2-Mikroglobulin. Ziel der Studie war es, die durch in vivo Bestimmung dieser Zytokine gewonnenen Ergebnisse mit bisher vorliegenden, meist in vitro gewonnenen Daten zu vergleichen und weitere Hinweise über den Wirkmechanismus von IFN-beta 1b zu erhalten. Systemic interferon-beta 1b (IFN-beta 1b)-therapy reduces significantly frequency and severity of relapses in multiple sclerosis (MS) patients. The details of the mechanism of action are still unknown; possibly the therapeutic effect could be mediated by downregulation of proinflammatory or by upregulation of antiinflammatory cytokines. 10 (pro-and antiinflammatory) cytokines have been investigated for this study via ELISA in the serum of 23 multiple sclerosis patients at six different points of time within a period of three months, it being: TNF-alpha, sTNF-R, IL-6, sIL-6R, Il-1beta, IL-1ra, sIL-2R, IL-10, neopterin, beta2-microglobulin. It has been the aim of this study to compare the results which have been gained by in vivo measurement of these cytokines with informations which had been gained previously by in vitro measurement. Another aim has been to learn more about the mechanism of action of IFN-beta 1b.

PURPOSE: The present study was designed to determine the normal range of serum beta 2-microglobulin (Sb2M) levels among healthy volunteers in Brazil. Levels of Sb2M are elevated in human immunodeficiency virus (HIV)-infected patients and have been shown to be the best predictor of HIV infection status and of some malignant disorders, especially multiple myeloma. In order to achieve its optimal use in Brazilian clinical diagnosis, an adequate reference interval study was performed for Sb2M IMMULITE® assay, based on the fact that its reference range limits were evaluated among European populations. METHODS: Ninety-six healthy blood donors were evaluated, and Sb2M levels were measured by chemiluminescent enzyme immunoassay using the IMMULITE® automated analyzer. RESULTS: A normal range of Sb2M values, established by a nonparametric statistical method, was 1.05 to 3.9 mg/mL, with the upper limit being higher than that reported elsewhere. CONCLUSIONS: This study presented new data indicating that there is a significant difference between the current reference limits for Sb2M IMMULITE® assay and those found in Brazil, providing evidence that significant differences in range of normal values may occur among different populations, and that these new values should be considered for Brazilian individuals. OBJETIVO: O objetivo do presente estudo foi determinar valores de referência de beta2-Microglobulina sérica (Sb2M) em voluntários saudáveis. Sabe-se que tal parâmetro apresenta-se elevado em pacientes infectados com o vírus da imunodeficiência humana (HIV) e tem se mostrado melhor marcador da infecção por HIV e de desordens malignas, especialmente mieloma múltiplo. De forma a se obter o melhor diagnóstico clínico, um intervalo de referência adequado de Sb2M para a população brasileira foi determinado empregando-se o ensaio IMMULITE®; já que este tem como parâmetro uma faixa de referência determinada a partir de populações européias. MÉTODOS: Noventa e seis doadores de sangue saudáveis foram avaliados e os valores de Sb2M foram medidos por método enzima imunoensaio quimiluminescente usando analisador automatizado IMMULITE®. RESULTADOS: Os valores de Sb2M, estabelecidos por método estatístico não paramétrico, apresentaram-se entre 1.05 e 3.9 mg/ml, sendo que o limite superior obtido foi maior que o relatado em outros trabalhos. CONCLUSÃO: Este estudo apresentou novos valores, indicando que existe uma diferença significante entre os limites de referência de Sb2M disponibilizados para o IMMULITE® e os encontrados no presente trabalho, evidenciando a ocorrência de variações importantes entre diferentes populações e que novos valores devem ser considerados para brasileiros.

PURPOSE: The present study was designed to determine the normal range of serum beta 2-microglobulin (Sb2M) levels among healthy volunteers in Brazil. Levels of Sb2M are elevated in human immunodeficiency virus (HIV)-infected patients and have been shown to be the best predictor of HIV infection status and of some malignant disorders, especially multiple myeloma. In order to achieve its optimal use in Brazilian clinical diagnosis, an adequate reference interval study was performed for Sb2M IMMULITE® assay, based on the fact that its reference range limits were evaluated among European populations. METHODS: Ninety-six healthy blood donors were evaluated, and Sb2M levels were measured by chemiluminescent enzyme immunoassay using the IMMULITE® automated analyzer. RESULTS: A normal range of Sb2M values, established by a nonparametric statistical method, was 1.05 to 3.9 mg/mL, with the upper limit being higher than that reported elsewhere. CONCLUSIONS: This study presented new data indicating that there is a significant difference between the current reference limits for Sb2M IMMULITE® assay and those found in Brazil, providing evidence that significant differences in range of normal values may occur among different populations, and that these new values should be considered for Brazilian individuals. OBJETIVO: O objetivo do presente estudo foi determinar valores de referência de beta2-Microglobulina sérica (Sb2M) em voluntários saudáveis. Sabe-se que tal parâmetro apresenta-se elevado em pacientes infectados com o vírus da imunodeficiência humana (HIV) e tem se mostrado melhor marcador da infecção por HIV e de desordens malignas, especialmente mieloma múltiplo. De forma a se obter o melhor diagnóstico clínico, um intervalo de referência adequado de Sb2M para a população brasileira foi determinado empregando-se o ensaio IMMULITE®; já que este tem como parâmetro uma faixa de referência determinada a partir de populações européias. MÉTODOS: Noventa e seis doadores de sangue saudáveis foram avaliados e os valores de Sb2M foram medidos por método enzima imunoensaio quimiluminescente usando analisador automatizado IMMULITE®. RESULTADOS: Os valores de Sb2M, estabelecidos por método estatístico não paramétrico, apresentaram-se entre 1.05 e 3.9 mg/ml, sendo que o limite superior obtido foi maior que o relatado em outros trabalhos. CONCLUSÃO: Este estudo apresentou novos valores, indicando que existe uma diferença significante entre os limites de referência de Sb2M disponibilizados para o IMMULITE® e os encontrados no presente trabalho, evidenciando a ocorrência de variações importantes entre diferentes populações e que novos valores devem ser considerados para brasileiros.

Cilj ovog istraživanja je bio odrediti izražaj gangliozida i neutralnih glikosfingolipida (GSL) u tkivima miševa kojima je izbačen gen za receptor TNF-a p55 (TNFR1-/-, od engl. tumor necrosis factor receptor, receptor za čimbenik nekroze tumora) te miševa kojima je izbačen gen za b2-mikroglobulin (b2MG-/-) koji je sastavni dio molekule MHC klase I. GSL su određeni u tkivima homozigotnih (TNFR1-/- i b2MG-/-) i heterozigotnih (TNFR1+/- i b2MG+/-) miševa za pojedinu gensku deleciju postupkom HPTLC (od engl. high perfomance thin layer chromatography, tankoslojna kromatografija visokog razlučivanja) te imunobojenjem GSL-kromatograma protutijelima na različite GSL. Oba modela imunodeficijentnih miševa (TNFR1-/- i b2MG-/-) su pokazala smanjen izražaj gangliozida tipa GM1a, GM1b i GM3 u plućima u odnosu na kontrolne skupine (TNFR1+/- i b2MG+/-). Rezultati ispitivanja miševa s delecijom gena za TNFR1 ukazuju da je prijenos pobudnog signala putem TNFR1 važan za stjecanje izražaja neutralnih GSL Gg3, Gg4, Gb4 i Gb5 u limfnim čvorovima ; Gg4 i Gb4 u slezeni te gangliozida tipa GM1b u limfoidnim tkivima (slezena, timus i limfni čvorovi). TNF također potiče izražaj gangliozida neolaktoserije u limfnim čvorovima a obrnuti učinak ima na izražaj pojedinih gangliozida neolaktoserije u tkivima mozga, pluća i mišića. Uz gangliozid IV3NeuAc-nLc4(C24) u mozgu TNFR1-/- bio je jače izražen i gangliozid Gb3. Delecija gena za b2MG uzrokuje specifično smanjenje izražaja neutralnih GSL Gg3, Gg4 i Gb3 u plućima. Neutralni GSL Gb4 je bio izraženiji u mozgu i limfnim čvorovima b2MG-/- u odnosu na kontrolne b2MG+/-miševe. Rezultati ovog doktorskog rada na in vivo modelima miševa bez gena za TNFR1 te miševa bez gena za b2MG dokazuju da miševi s genskom delecijom molekula nužnih za normalnu imunološku reakciju mogu promijeniti izražaj GSL u limfatičkim i drugim tkivima.

Beta2-microglobulin is a constituent of the class I major histocompatibility complex (MHC) molecule and crucial for its normal function in cell recognition.It has also been isolated from bone and shown to regulate bone metabolism and to be altered in various bone diseases.In order to further investigate the role of the immune system in bone metabolism, we studied basic properties of bone physiology in beta2-microglobulin-deficient mice created by the tehnique of gene knock-out.Ten week- old male offspring homozygous (non-functional class I MHC molecule) or heterozygous (functional class I MHC molecule) for beta2-microglobulin knock-out gene did not differ in the following measures of bone turnover: femur lenght, dry and ash weight and calcium content, serum calcium concentration and alkaline phosphatase activity, total vertebral tissue area, trabecular bone volume, osteid surface, osteoclast surface and mineral apposition rate.These data indicate that the bone turnover in beta2- microglobulin-deficient mice is appropriate for the stage of their skeletal maturation.

Expression of beta2-microglobulin (β2m) in the common carp was studied using a polyclonal antibody raised against a recombinant protein obtained from eukaryotic expression of theCyca-B2mgene.β2m is expressed on peripheral blood Ig+and Ig-lymphocytes, but not on erythrocytes and thrombocytes. In spleen and pronephros, dull- and bright-positive populations could be identified correlating with the presence of erythrocytes, thrombocytes, and mature leucocytes or immature and mature cells from the lympho-myeloid lineage, respectively. Thymocytes were shown to be comprised of a single bright-positive population. The Cyca-B2m polyclonal antiserum was used in conjunction with a similarly produced polyclonal antiserum to an MHC class I (Cyca-UA)αchain to investigate the expression of class I molecules on peripheral blood leucocytes (PBL) at different permissive temperatures. At 12℃, a temporary downregulation of class I molecules was demonstrated, which recovered to normal levels within 3 days. However, at 6℃, a lasting absence of class I cell-surface expression was observed, which could be restored slowly by transfer to 12C. The expression of immunoglobulin molecules on B cells was unaffected by temperature changes. The absence of the class cell-surface expression was shown to be the result of a lack of sufficientCyca-B2mgene transcription, althoughCyca-UAmRNA was present at comparable levels at all temperatures. This suggests that class I expression is regulated by a temperature-sensitive transcription of theCyca-B2mgene.

(Full text is available at http://www.manu.edu.mk/prilozi). Endemic nephropathy (EN) is a chronic tubulointerstitial nephropathy with an early insidious and slow development into terminal renal failure. Proteomics is the systematic study of a proteome, which is the total protein content of a cell, organism or body fluids. Application of proteomic technologies in nephrology has enabled more detailed analyses of protein functions and examined their importance in various physiological and pathological states. Biomarkers with high specificity and sensitivity to early diagnosis are needed for a better understanding of the mechanisms of EN development and its consequences. Urine beta2-microglobulin (B2M) was mainly used as a tubular marker of EN but recently alpha1-microglobulin (AMBP) was proposed for the diagnosis of EN. We studied the urine proteins of 360 patients with EN, diabetic nephropathy (DN) and acute kidney injury (AKI) and the healthy population using proteomic tools. Protein maps from the urine of patients with EN showed significant differences in comparison to the healthy subjects and patients with DN and AKI. Our study highlights six proteins in urine that were differentially excreted in the urine of EN patients compared with the other groups and have potential to be markers for EN prediction. In one of our studies, using routine biomarkers, we investigated the potential of urine B2M, AMBP, albumin and total protein as diagnostic markers for EN, in comparison to glomerulonephritis, nephrosclerosis and a healthy state. Modern proteomic technologies are still robust investigation tools, but can access a vast amount of information from one set of experiments in comparison to a classic diagnostic approach. Key words: endemic nephropathy, beta2-microglobulin, alpha1-microglobulin, urine proteomics. peerReviewed

Background: chronic kidney disease (CKD) increases the severity and risk of mortality in acute coronary syndrome (ACS) patients. The role of β2-M as a filtration and inflammation marker and FGF23 as a CKD-MBD process marker might be significant in the pathophysiology in ACS with CKD patients. This study aims to determine the association of β2-M and FGF23 with major adverse cardiac event (MACE) in ACS patients with CKD. Methods: we used cross sectional and retrospective cohort analysis for MACE. We collected ACS patients with CKD consecutively from January until October 2018 at Dr. Cipto Mangunkusumo General Hospital. Data were analyzed using logistic regression and Cox’s Proportional Hazard Regression. Results: a total of 117 patients were selected according to the study criteria. In bivariate analysis, β2-M, FGF23, and stage of CKD had significant association with MACE (p = 0.014, p = 0.026, p = 0.014, respectively). In multivariate analysis, β2-M - but not FGF 23- was significantly associated with MACE (adjusted HR 2.16; CI95% 1.15–4.05; p = 0.017). Conclusion: β2-M was significantly associated with MACE, while FGF23 was not so. This finding supports the role of inflammation in cardiovascular outcomes in ACS with CKD patient through acute on chronic effect.