Screening for urinary markers predicting hematopoietic stem cell injury induced by busulfan using genetically diverse mice

Background:Busulfan(BU)is an alkylating agent used as a conditioning agent prior to hematopoietic stem cell(HSC)transplantation as it is known to be cytotoxic to host hematopoietic stem and progenitor cells.The susceptibility of HSCs to BU injury plays an important role in the myeloablative efficacy of BU.Different susceptibilities were demonstrated in genetically diverse(GD)mice in our preliminary research.Methods:Three strains of GD mice with different susceptibilities to BU-i nduced HSC injury were used for screening biological markers of HSC injury susceptibility in urine.The urine proteins were analyzed using liquid chromatography coupled with tandem mass spectrometry to screen for differentially expressed proteins.Screening for possible biomarkers based on differences in protein expression abundance was validated using enzyme-l inked immunoassay(ELISA).Results:Functional analysis showed that the differential proteins were all involved in a series of biological pathways related to cellular senescence,apoptosis,and angiogenesis;whereas the differential proteins of the high-susceptible strain were enriched for the regulation of bone marrow microenvironment pathways,those of low-susceptible strain were enriched for the proapoptotic effect of GTPase pathways.Based on protein abundance differences,several urinary proteins that may be indicative of susceptibility were screened,and ELISA validation results showed that angiotensin-converting enzyme may be a potential biomarker predicting HSC susceptibility for BU conditioning.Conclusions:This study indicates that urinary protein levels can reflect differences in susceptibility to BU-i nduced HSC injury.Using GD mice to construct genetic difference models will provide preclinical data for screening BU-related biological markers.

Neutrophil gelatinase-associated lipocalin,kidney injury molecule-1,and periostin:Novel urinary biomarkers in diabetic nephropathy

BACKGROUND Globally,diabetic nephropathy(DN)is the primary cause of chronic kidney disease.Currently,renal function is monitored indirectly using measures of serum creatinine,estimated glomerular filtration rate(eGFR),and proteinuria.Novel urinary biomarkers utilized in the early stages of DN have been described;these indicators can be used in the early identification of the disease,which is important for initiating treatment to halt or impediment the advance of diabetic nephropathy.AIM To estimate neutrophil gelatinase-associated lipocalin(NGAL),kidney injury molecule-1(KIM-1),and periostin(POSTN)levels as novel urinary biomarkers in DN.METHODS In this hospital based cross-sectional study,a total of 160 patients of both genders aged 18 years or more;40 healthy participants and 120 patients with diabetes mellitus(DM)were included.Patients with DM were divided into normoalbuminuria(n=40),microalbuminuria(n=40),and macroalbuminuria(n=40)groups as per urine albumin creatinine ratio(uACR).Blood urea,serum creatinine,uACR were measured.Urine NGAL,KIM-1,and POSTN were measured by enzyme linked immunosorbent assay.The eGFR was calculated and compared with urinary markers.RESULTS NGAL,KIM-1,and POSTN levels increased significantly in normo,micro,and macroalbuminuria with the highest in the macroalbuminuria group.Albumin creatinine ratio(ACR)showed a positive correlation with NGAL,KIM-1,and POSTN levels.The eGFR showed a weak negative correlation with ACR,NGAL,KIM-1,and POSTN.NGAL was significantly lower in stage 1 compared to stage 2,3,and 4 kidney disease.KIM-1 was significantly decreased in stage 1 compared to stage 4 kidney disease.POSTN was significantly decreased in stage 1 compared to stage 3 and 4 kidney disease.The receiver operator curve analysis of ACR,NGAL,KIM-1,and POSTN showed good sensitivity of 80%,75.8%,63.3%,and 80%respectively with a cut-off of 12.5 mg/g,4.5μg/L,1.5 ng/mL,and 37.5 ng/mL.CONCLUSION Urinary NGAL and POSTN are independent markers of DN.

Early Results of Urothelial Carcinoma Screening in a Risk Population of Coke Workers:Urothelial Carcinoma among Coke Workers

Objective To present the protocol and the early results of a urothelial carcinoma （UC） screening analysis performed in a risk population of coke workers. Methods Between June 2006 and October 2008, 171 male workers （mean age 43 years）, employed in a Ligurian coke plant （Italiana Coke S.r.l） and exposed to polycyclic aromatic hydrocarbons （PAHs） for a median period of 16 years, underwent screening for UC. Urological evaluation included medical history, physical examination, routine laboratory tests, urine analysis, urinary cytology and uCyt＋ assay. In the event of signs and symptoms suggestive of UC or positive urinary tests, the workers were also subjected to urinary ultrasonography and cystoscopy with biopsy of any suspicious lesions. Results Regarding the laboratory tests, 19/171 （11%） uCyt＋ samples were considered inadequate and were excluded from the outcomes assessment. Overall, urine analysis, cytology and uCyt＋ were positive in 18/152 （12%） subjects who showed no evidence of UC at the scheduled check-ups. No significant association was identified between marker positivity and occupational activity. Conclusions Our results fail to show an increased risk of UC among the coke workers evaluated. However, they will need to be confirmed in the future by a larger enrollment and a longer follow-up in order to assess the definitive risk for UC after exposure to coke.