An updated catalog of lipocalins of the chagas disease vector Rhodnius prolixus (Hemiptera, Reduviidae).

The haematophagy process by arthropods has been one of the main targets of studies in the parasite-host interaction, and the kissing-bug Rhodnius prolixus, vector of the protozoan Trypanosoma cruzi, has been one of the main models for such studies. Still in the 1980s, it was identified that among the salivary proteins for disrupting vertebrate host homeostasis, lipocalins were among the most relevant proteins for this process. Since then, 30 lipocalins have been identified in the salivary glands of R. prolixus, that promotes vasodilatation, prevents inflammation, act as anticoagulants and inhibits platelet aggregation. The present work aims to identify new lipocalins from R. prolixus, combining transcriptome and genome data. Identified new genes were mapped and had their structure annotated. To infer an evolutionary relationship between lipocalins, and to support the predicted functions for each lipocalin, all amino acid sequences were used to construct phylogenetic trees. We identified a total of 29 new lipocalins, 3 new bioaminogenic-biding proteins (which act to inhibit platelet aggregation and vasodilation), 9 new inhibitors of platelet aggregation, 7 new apolipoproteins and 10 lipocalins with no putative function. In addition, we observed that several of the lipocalins are also expressed in different R. prolxius tissues, including gut, central nervous system, antennae, and reproductive organs. In addition to newly identified lipocalins and a mapping the new and old lipocalins in the genome of R. prolixus, our study also carried out a review on functional status and nomenclature of some of the already identified lipocalins. Our study reinforces that we are far from understanding the role of lipocalins in the physiology of R. prolixus, and that studies of this family are still of great relevance.

The role of urinary KIM-1, NGAL, CA19-9 and ß2-microglobulin in the assessment of ureteropelvic junction obstruction in adults.

PURPOSE: The objective of this study is to evaluate the diagnostic properties of urinary biomarkers in adults with ureteropelvic junction obstruction: KIM-1, NGAL, CA19-9, and ß2-microglobulin. We also assessed urinary biomarker concentrations following pyeloplasty. MATERIAL AND METHODS: We prospectively studied adults from December 2013 to February 2015. We included 47 patients with a mean age of 38.6 ± 12.7 years. Each patient provided four samples of voided urine for biomarker measurement, one at pre-operative consultation and the others at 1, 3, and 6 months of post-operative follow-up. The control group consisted of 40 healthy individuals with no hydronephrosis on ultrasound evaluation. RESULTS: KIM-1 had an area under the curve of 0.79 (95% CI 0.70-0.89), NGAL 0.71 (95% CI 0.61-0.83), CA19-9 0.70 (95% CI 0.60-0.81), and ß2-microgloblin 0.61 (95% CI 0.50-0.73). KIM-1 was the most sensitive marker with a cut-off of 170.4 pg/mg creatinine (sensitivity 91.4%, specificity 59.1%), whereas CA19-9 was the most specific with a cut-off of 51.3 U/mg creatinine (sensitivity 48.9%, specificity 88.0%). Urinary concentrations of biomarkers decreased after pyeloplasty. CONCLUSIONS: The evaluation of urinary biomarkers is useful in adults undergoing pyeloplasty. KIM-1, NGAL, and CA19-9 were elevated and significantly decreased after surgery.

NGAL urinária em pacientes sem e com lesão renal aguda em unidade de terapia intensiva / Urinary NGAL in patients with and without acute kidney injury in a cardiology intensive care unit

Objetivo: Avaliar a eficácia diagnóstica e prognóstica da lipocalina associada à gelatinase neutrofílica urinária em pacientes de unidade de terapia intensiva. Métodos: Estudo do tipo coorte, prospectivo, longitudinal desenvolvido em uma unidade de terapia intensiva clínica especializada em cardiologia. Os pacientes foram estratificados segundo os grupos sem e com lesão renal aguda, acompanhados a partir das primeiras 24 horas de internação até a alta hospitalar ou óbito. A creatinina sérica, o fluxo urinário e a lipocalina associada à gelatinase neutrofílica urinária foram coletadas em dois períodos: 24 horas e 48 horas de admissão. Resultados: Foram avaliados 83 pacientes clínicos da unidade de terapia intensiva, com predomínio do gênero masculino (57,8%). Os pacientes foram agrupados em sem lesão renal aguda (N=18), com lesão renal aguda (N=28) ou com lesão renal aguda grave (N=37). Entre os pacientes com lesão renal aguda e lesão renal aguda grave, foram prevalentes os portadores de doenças crônicas, em uso de ventilação mecânica e em terapia de substituição renal, além daqueles com maiores taxas de permanência na unidade de terapia intensiva e hospitalar, e maior mortalidade. O grupo com lesão renal aguda não apresentou alteração significativa da creatinina sérica nas primeiras 24 horas na unidade de terapia intensiva, ...

Objective: To assess the diagnostic and prognostic efficacy of urine neutrophil gelatinase-associated lipocalin in patients admitted to an intensive care unit. Methods: Longitudinal, prospective cohort study conducted in a cardiology intensive care unit. The participants were divided into groups with and without acute kidney injury and were followed from admission to the intensive care unit until hospital discharge or death. Serum creatinine, urine output and urine neutrophil gelatinase-associated lipocalin were measured 24 and 48 hours after admission. Results: A total of 83 patients admitted to the intensive care unit for clinical reasons were assessed, most being male (57.8%). The participants were divided into groups without acute kidney injury (N=18), with acute kidney injury (N=28) and with severe acute kidney injury (N=37). Chronic diseases, mechanical ventilation and renal replacement therapy were more common in the groups with acute kidney injury and severe acute kidney injury, and those groups exhibited longer intensive care unit stay and hospital stay and higher mortality. Serum creatinine did not change significantly in the group with acute kidney injury within the first 24 hours of admission to the intensive care unit, although, urine neutrophil gelatinase-associated lipocalin was high in the groups with acute kidney injury and severe acute kidney injury (p<0.001). Increased urine neutrophil gelatinase-associated lipocalin was associated with death. ...

Papel da lipocalina associada à gelatinase neutrofílica (NGAL) urinária na nefrotoxicidade da cisplatina em pacientes com câncer de cabeça e pescoço / Evaluation of the cisplatin nephrotoxicity using the urinary neutrophil gelatinase-associated lipocalin (NGAL) in patients with head and neck cancer

Introdução: A injúria renal aguda (IRA) em pacientes que recebem a cisplatina é comum, portanto, a avaliação da função renal em pacientes utilizando drogas nefrotóxicas é fundamental. Objetivo: Avaliar a incidência da IRA e o papel da lipocalina associada à gelatinase neutrofílica (NGAL) na avaliação da função renal em pacientes com câncer de cabeça e pescoço (CCP) que receberam a cisplatina. Métodos: Foram avaliados prospectivamente 50 pacientes com CCP, tratados com três sessões de cisplatina. Foram coletados sangue e urina 24 horas antes da cisplatina, 24 horas após a infusão, 48 horas após cada aplicação e 35 dias após o término do tratamento (NGAL urinária, proteína C reativa, creatinina e taxa de filtração glomerular, desidrogenase lática e magnésio plasmáticos). Resultados: A IRA foi observada em 78% dos pacientes. Houve aumento na creatinina, ureia e queda na TFG após cada ciclo de cisplatina, e aumento da NGAL urinária. Foi observada associação positiva entre os níveis de NGAL e a creatinina e PCR. Evidenciou-se um aumento dos níveis de creatinina, NGAL, PCR e diminuição da TFG nos pacientes com IRA em relação aos pacientes sem IRA. Conclusão: Observamos IRA em 78% dos pacientes avaliados com CCP tratados com a cisplatina e correlação da NGAL com a creatinina e a TFG em demonstrar lesão renal. Os níveis de NGAL podem estar elevados em relação aos níveis basais, mesmo antes da utilização da cisplatina. .

Introduction: Acute kidney injury (AKI) in patients receiving cisplatin is common, therefore the evaluation of renal function in patients on use of nephrotoxic drugs is fundamental. Objective: To evaluate the incidence of AKI and the role of lipocalin associated to neutrophil gelatinase (NGAL) in the monitoring of renal function in patients with head and neck cancer (HNC) who received cisplatin. Methods: We prospectively studied 50 patients with HNC treated with three sessions of cisplatin. Blood and urine were collected 24 hours before cisplatin, 24 hours after infusion, 48 hours after each application and 35 days after the end of treatment (urine NGAL, C-reactive protein, creatinine, glomerular filtration rate, plasma lactate dehydrogenase and magnesium). Results: AKI was observed in 78% of patients. There was increase in creatinine, and decrease in GFR after each cycle of cisplatin, and increased urine NGAL. Positive association was observed between the levels of NGAL, creatinine and C-reactive protein. It was observed an increase in creatinine, NGAL, C-reactive protein and decreased GFR in AKI patients compared to patients without AKI. Conclusion: AKI was noted in 78% of patients with HNC treated with cisplatin and showed the correlation of NGAL with creatinine and GFR in demonstrating renal injury. NGAL levels may be elevated compared to baseline levels, even before the use of cisplatin. .

Overexpression of a monomeric form of the bovine odorant-binding protein protects Escherichia coli from chemical-induced oxidative stress.

Mammalian odorant-binding proteins (OBPs) are soluble lipocalins produced in the nasal mucosa and in other epithelial tissues of several animal species, where they are supposed to serve as scavengers for small structurally unrelated hydrophobic molecules. These would include odorants and toxic aldehydes like 4-hydroxy-2-nonenal (HNE), which are end products of lipid peroxidation; therefore OBP might physiologically contribute to preserve the integrity of epithelial tissues under oxidative stress conditions by removing toxic compounds from the environment and, eventually, driving them to the appropriate degradative pathways. With the aim of developing a biological model based on a living organism for the investigation of the antioxidant properties of OBP, here we asked whether the overexpression of the protein could confer protection from chemical-induced oxidative stress in Escherichia coli. To this aim, bacteria were made to overexpress either GCC-bOBP, a redesigned monomeric mutant of bovine OBP, or its amino-terminal 6-histidine-tagged version 6H-GCC-bOBP. After inducing overexpression for 4 h, bacterial cells were diluted in fresh culture media, and their growth curves were followed in the presence of hydrogen peroxide (H2O2) and tert-Butyl hydroperoxide (tBuOOH), two reactive oxygen species whose toxicity is mainly due to lipid peroxidation, and menadione, a redox-cycling drug producing the superoxide ion. GCC-bOBP and 6H-GCC-bOBP were found to protect bacterial cells from the insulting agents H2O2 and tBuOOH but not from menadione. The obtained data led us to hypothesize that the presence of overexpressed OBP may contribute to protect bacterial cells against oxidative stress probably by sequestering toxic compounds locally produced during the first replication cycles by lipid peroxidation, before bacteria activate their appropriate enzyme-based antioxidative mechanisms.

Changes in various endometrial proteins during cloprostenol-induced failure of early pregnancy in mares

Various major proteins in uterine secretions flushed from mares on day 20 of gestation have been identified and related to levels of gene expression in endometrial biopsies. Pregnancy-related changes were determined by comparison with samples from mares on day 20 after being given a luteolytic IM dose of cloprostenol on day 18, and with cycling mares that were not pregnant on days 17-20. Proteins in cell-free uterine flush samples were identified by LC-MS/MS of trypsin- digested peptides. Levels of endometrial expression were estimated from raw signal strength in Agilent 21322 E. caballus expression microarray. In this paper we describe a subset of pregnancy-related endometrial proteins with potential roles in embryonic development and/or mucosal innate immunity. Endometrial proteins that increased greatly during pregnancy included GM2 activator (GM2A), lipocalin 2 (LCN2), stanniocalcin 1 (STC1) and uterine serpin (SERPINA14). Endometrial proteins that decreased in normal pregnancy included secretory phospholipase A2 (sPLA2), secretoglobin 1A1 (SCGB1A1), and vanin 1 (VNN1). Cloprostenol-induced failure of pregnancy resulted in increased endometrial expression of SPLA2, cathepsin L1 (CTSL1), interleukin-1 receptor antagonist (IL1RN), SCGB1A1 epididymal secretory protein E1 (NPC2), connective tissue growth factor (CTGF), retinol binding protein 4 (RBP4), uteroferrin/TRAP5 (ACP5), SERPINA14, annexin A1 (ANXA1), annexin A3 (ANXA3), and vanin 3 (VNN3). Expression of P19 uterocalin (P19) was similarly high in pregnant and non-pregnant endometrium, and virtually undetectable in the yolk-sac wall. High levels of expression of ANXA2, GM2A, NPC2, CTSL1 and RBP4 in both endometrium and yolk-sac wall suggests that these have important roles on both sides of the maternal:conceptus interface. The properties and distribution of GM2A and NPC2 suggest that they are important in the transport of phospholipids and sterols. SPLA2, LCN2, SCGB1A1 and IL1RA have potential roles in mucosal innate immunity but their production is reduced during early pregnancy. These studies provide an inventory of many proteins in the uterine lumen during early pregnancy, and a subset for which local endometrial expression is altered when pregnancy is compromised by cloprostenol-induced luteolysis.(AU)

Changes in various endometrial proteins during cloprostenol-induced failure of early pregnancy in mares

Various major proteins in uterine secretions flushed from mares on day 20 of gestation have been identified and related to levels of gene expression in endometrial biopsies. Pregnancy-related changes were determined by comparison with samples from mares on day 20 after being given a luteolytic IM dose of cloprostenol on day 18, and with cycling mares that were not pregnant on days 17-20. Proteins in cell-free uterine flush samples were identified by LC-MS/MS of trypsin- digested peptides. Levels of endometrial expression were estimated from raw signal strength in Agilent 21322 E. caballus expression microarray. In this paper we describe a subset of pregnancy-related endometrial proteins with potential roles in embryonic development and/or mucosal innate immunity. Endometrial proteins that increased greatly during pregnancy included GM2 activator (GM2A), lipocalin 2 (LCN2), stanniocalcin 1 (STC1) and uterine serpin (SERPINA14). Endometrial proteins that decreased in normal pregnancy included secretory phospholipase A2 (sPLA2), secretoglobin 1A1 (SCGB1A1), and vanin 1 (VNN1). Cloprostenol-induced failure of pregnancy resulted in increased endometrial expression of SPLA2, cathepsin L1 (CTSL1), interleukin-1 receptor antagonist (IL1RN), SCGB1A1 epididymal secretory protein E1 (NPC2), connective tissue growth factor (CTGF), retinol binding protein 4 (RBP4), uteroferrin/TRAP5 (ACP5), SERPINA14, annexin A1 (ANXA1), annexin A3 (ANXA3), and vanin 3 (VNN3). Expression of P19 uterocalin (P19) was similarly high in pregnant and non-pregnant endometrium, and virtually undetectable in the yolk-sac wall. High levels of expression of ANXA2, GM2A, NPC2, CTSL1 and RBP4 in both endometrium and yolk-sac wall suggests that these have important roles on both sides of the maternal:conceptus interface. The properties and distribution of GM2A and NPC2 suggest that they are important in the transport of phospholipids and sterols. SPLA2, LCN2, SCGB1A1 and IL1RA have potential roles in mucosal innate immunity but their production is reduced during early pregnancy. These studies provide an inventory of many proteins in the uterine lumen during early pregnancy, and a subset for which local endometrial expression is altered when pregnancy is compromised by cloprostenol-induced luteolysis.