MICROALBUMINURIA BESIDES TO URINARY ENZYMATIC PROTEIN LEVELS INCREASE IN DIABETIC KIDNEY DISEASE WITH TYPE II DIABETICS

Background: Diabetic kidney disease (DKD) is a time progressive problem, give rise in uncontrolled Diabetics increasing risks for chronic kidney disease (CKD) and /or end-stage renal disease (ESRD). The vulnerability to renal dysfunction manifested with sudden glomerular hypofiltration associated with micro-to macroalbuminuria passing to renal failure. So that, screening of specific enzymes shifts, or urinary albumin may predict onset diabetic nephropathy. Objective:The assessment of urinary alkaline phosphatase (ALP), alanine aminopeptidase (AAP), acid phosphatase (ACP) and microalbuminuria (MAU) for type II diabetic patients. Patients and Methods: In this study,120 type II diabetic patients were compared to 90 healthy volunteers of matched age and sex in Al-Leith General Hospital, Al-Leith Kidney Unit (AKU), Al-Leith, Makkah area, KSA in which random urine samples were collected for testing of MAU, ALP, AAP, ACP and Cr. Results: Mean values of measured biomarkers in patient group for MAU, ALP, AAP, ACP and Cr were 51.92 mg/I, 41.55 U/L, 20.17 U/L, 570.10 U/L and 2.92 mg/dl VS in control group were 12.59 mg/I, 8.84 U/L, 6.94 U/L, 385.87U/L and 1.07 mg/dl respectively. Additionally, there were statistically positive correlation between AAP with MAU and ALP; ACP with MAU, ALP and AAP; Cr level with MAU, ALP, AAP and ACP; on the other hand, there were positive significant correlation between duration of diabetes with all studied markers. Conclusion: Using of MAU in addition to other urinary enzymes could be beneficial non-invasive indicators for renal deterioration in type II diabetics

The role of Klotho and CYR61 proteins in early diagnosis of acute kidney injury after cardiac surgery

Acute kidney injury (AKI) is a critical problem accompanied with cardiac surgery as one of the adverse complications that contribute to prolonged hospitalization period and/or increase morbidity and mortality rates, surpassing 40% among cardiac ICU patients. Cardiac surgery with cardiopulmonary bypass (CPB) reproduces globally every year leading to volume depletion as a result of cardiac output exhaustion and stimulation of renin-angiotensin system with renal vasoconstriction that increases the extent of AKI among hospitalized patients. It is well known that AKI may worsen kidney function by time depending on slowly traditional biomarker “serum creatinine” which tardily increases after 2 days of renal tubular injury indicating that 50% of kidney function is lost and creatinine kinetics is influenced by ethnicity, gender, age, dietary protein intake, muscle mass and medications that didn’t reflect precisely the prognosis of AKI over time. Therefore, the needs for novel biomarkers are important situations to improve sensitivity, specificity of clinical diagnosis and potentiate the efficiency of treatment to prevent irreversible consequences. In the present doctoral work, the following hypotheses will be tested: 1) Plasma/serum or urinary levels of Klotho and CYR61 are early prognostic biomarkers of AKI after cardiac surgery. 2) Klotho and CYR61 are markers of renal tubular dysfunction at the different levels of AKI after cardiac surgery. 3) The combinations of Klotho and CYR61 with other markers (f.e. NGAL/Klotho or KIM-1/Klotho ratios) will increase the prognostic efficiency of AKI after cardiac surgery. Our study covered 50 patients admitted to cardiac surgery department in the University Medical Center Ljubljana and fulfilled the inclusion criteria of selection. The classification of patients into AKI group and non-AKI group relies on KDIGO (Kidney Disease Improving Global Outcomes) that declared its validation for AKI staging and severity in the recent ADAQI consensus, retained almost advantages of both RIFLE and AKIN criteria in a harmonized manner. Serum levels of creatinine, cystatin C, CYR61 and klotho were measured at five time intervals before and within 48 hours of CPB.Akutna ledvična okvara (ALO) je resen problem pri operacijah srca kot eden od zapletov, ki podaljšajo hospitalizacijo in/ali zvečajo obolevnost in smrtnost, ki pri srčnih bolnikih v enotah intenzivne nege (EIN) presegata 40 %. Operaciji srca s kardiopulmonalnim obvodom (KPO) prispevajo k zmanjšanju volumna, skupaj s stimulacijo sistema renin-angiotenzin in renalno vazokonstrikcijo zvečajo obseg ALO pri hospitaliziranih bolnikih. Znano je, da lahko ALO posledično poslabša ledvično funkcijo. Uveljavljen biooznačevalec serumski kreatinin se zviša šele po dveh dneh od tubulne okvare, ko ledvična funkcija upade za 50 %. Na kinetiko tega označevalca močno vpliva etnična pripadnost, spol, starost, vnos beljakovin, mišična masa in zdravila. Da bi preprečili ireverzibilne posledice, so potrebni novi biooznačevalci, ki bi izboljšali občutljivosti in specifičnosti klinične diagnoze in zvečali učinkovitost zdravljenja. V tej doktorski dela bodo testirali naslednje hipoteze: 1) Plazma / serum ali ravni sečil z Klotho in CYR61 so zgodnji napovedni biomarkerjev ALO po operaciji srca. 2) Klotho in CYR61 so označevalcev ledvično tubularno disfunkcijo na različnih ravneh ALO po operaciji srca. 3) kombinacije Klotho in CYR61 z drugimi označevalci (npr. NGAL / Klotho ali KIM-1 razmerja / Klotho) se bo povečala napovedni učinkovitost ALO po operaciji srca. Naša raziskava je vključevala 50 bolnikov z načrtovano operacijo srca, ki so bili sprejeti na oddelek srčne kirurgije v Univerzitetnem kliničnem centru Ljubljana in so izpolnjevali vključitvene kriterije. Bolniki so bili razvrščeni v ALO in brez ALO skupino glede na kriterij KDIGO (Kidney Disease Improving Global Outcomes), ki je bil potrjen za oceno stopnje ALO z nedavnim ADAQI konsenzom in združuje večino prednosti RIFLE in AKIN kriterijev. Merili smo serumski nivo kreatinina, cistatina C, CYR61 in Klotho pri petih časovnih intervalih pred operacijo in 48 ur po CPB. Meritev cistatina C skupaj s kreatininom naj bi bila koristna iz več vidikov. Nivoja cistatina C in kreatinina sta pomembna pri oceni GFR z MDRD formulo, ob upadu GFR nivo cistatina C naraste ledvicah, urinu in periferni krvi. Poleg tega naj bi bil nivo cistatina C pred operacijo in en dan po operaciji pokazatelj razvoja ALO. V naši raziskavi sta se serumska nivoja kreatinina in cistatina C podobno spreminjala v ALO in brez ALO skupini. V ALO skupini sta bila oba zvišana nad začetno vrednost dve uri po CPB in sta dosegla najvišjo vrednost drugi dan po CPB. V brez ALO skupini sta se oba nivoja v začetku hitro znižala, zatem pa se je potek razlikoval: kreatinin se je zvišal po enem dnevu in po dveh dneh upadel na izhodiščno vrednost, cistatin C pa je ostal zvišan nad izhodiščno vrednostjo tudi po dveh dneh. Vloga CYR61 v telesu še ni popolnoma poznana. Sodeluje pri celjenju ran, fibrozi, vnetju, artritisu, žilnih boleznih in raku. Pri boleznih ledvic so preučevanja nivoja v urinu in meritev nivoja mRNA dala spodbudne rezultate, nivoja v krvi pa še niso preučevali. V raziskavi smo v tem kontekstu prvi preučevali serumski nivo humanega CYR61. Višje začetne vrednosti smo izmerili v brez ALO skupini. Po dveh dneh po CPB so vrednosti v brez ALO upadle na polovico začetne vrednosti, medtem ko so bile v ALO skupini rahlo - vendar ne statistično značilno - višje od izhodiščnih. Nivo CYR61 bi lahko bil uporaben za razlikovanje stopenj I in II v ALO skupini, a ta povezava je zaradi omejenega števila bolnikov v naši raziskavi dosegla le mejno statistično značilnost. Klotho ima v telesu razne vloge: pri procesih staranja, kot antagonist oksidativnega stresa in delovanje skupaj s hormoni, kot je FGF-23. Sprošča se v raznih tkivih, tudi v ledvičnih tubulih. Z raziskavami na živalih so ugotovili, da se po ishemični poškodbi ledvic zmanjša nastajanje mRNA in tudi nivo Klotho v urinu in plazmi. V naši raziskavi je bil izhodiščni nivo Klotho višji v ALO kot v brez ALO skupini, razlika med skupinama je bila podobna tudi dve uri po CPB. Vrednosti so se znižale dva dni po operaciji. Gibanje nivoja merjenih biooznačevalcev kaže, da imata Klotho in cistatin C pri ALO večji potencial kot CYR61. Kombinacija Klotho in kreatinina pa bi lahko še zvečala diagnostično uporabnost pri ALO. Na koncu smo glede na naše rezultate lahko potrdili prvo in tretjo hipotezo ter delno tudi drugo

THE PROTECTIVE ROLE OF TANNIC ACID AGAINST POSSIBLE HEPATO-NEPHROTOXICITY INDUCED BY SILVER NANOPARTICLES ON MALE RATS

Silver nanoparticles (AgNPs) are being used extensively for biomedical purposes regarding to their broad antimicrobial activity, however their toxicity has been addressed in only few studies. In the present study, we aimed to prepare and characterize AgNPs, investigate their adverse effect on liver and kidney functions, and also elucidate the hepato-nephro protective ability of tannic acid in male rats. The obtained results showed that AgNPs caused oxidative stress throughout the induction of thiobarbituric acid-reactive substances (TBARS) and the reduction of the activities of antioxidant enzymes (GST, SOD, CAT, GPx) and the levels of glutathione. Hepatic markers enzymes (AST, ALT, ALP, ACP, LDH and GGT), total bilirubin, urea, creatinine and lipid profile were increased, while hematological parameters were decreased. Histopathological investigations indicated marked degeneration of hepatocytes, endothelial cells of renal which with its role has confirmed the hepatotoxicity and nephrotoxicity induced by AgNPs. The presence of tannic acid along with AgNPs showed obvious improvements in the injured liver and kidney tissues. The protective effect of tannic acid against the toxicity of AgNPs might be due to its antioxidant properties and scavenging abilities against active free radicals

Serum Klotho as a marker for early diagnosis of acute kidney injury after cardiac surgery

Background: Early diagnosis of acute kidney injury (AKI) after cardiac surgery is based on serum creatinine which is neither a specific nor a sensitive biomarker. In our study, we investigated the role of serum Klotho in early prediction of AKI after cardiac surgery using cardiopulmonary bypass (CPB). Methods: The included patients were classified into three groups according to AKI stages using KDIGO criteria. The measurements of creatinine and Klotho levels in serum were performed before surgery, at the end of CPB, 2 hours after the end of CPB, 24 hours and 48 hours postoperatively. Results: Seventy-eight patients were included in the study. A significant increase of creatinine levels (p<0.001) was measured on the first day after the surgery in both AKI groups compared to the non-AKI group. However, a significant difference between AKI-2 and AKI-1 groups (p=0.006) was not measured until the second day after the operation. Using decision trees for classification of patients with a higher or lower risk of AKI we found out that Klotho discriminated between the patients at low risk of developing more severe kidney injury in the first hours after surgery and the patients at high risk better than creatinine. Adding also the early measurements of creatinine in the decision tree model further improved the prediction of AKI. Conclusions: Serum Klotho may be useful to discriminate between the patients at lower and the patients at higher risk of developing severe kidney injury after cardiac surgery using CPB already in the first hours after surgery.Uvod: Rana dijagnoza akutnog oštećenja bubrega (AKI) posle operacije srca se zasniva na kreatininu u serumu koji nije ni specifičan ni osetljiv biomarker. U našoj studiji, istraživali smo ulogu serumskog Klotho-a u ranom predvi|anju AKI nakon operacije srca koristeći kardiopulmonarni bajpas (CPB). Metode: Uključeni pacijenti su klasifikovani u tri grupe u skladu sa AKI fazama po KDIGO kriterijumu. Merenje kreatinina i Klotho-a u serumu je bilo izvedeno pre operacije, krajem CPB, dva sata nakon završetka CPB, 24 sati i 48 sati postoperativno. Rezultati: U studiju je uključeno sedamdeset osam pacijenata. Povišenje nivoa kreatinina prvi dan posle operacije je bilo značajno (p < 0,001) u obe AKI grupe u pore|enju sa non-AKI grupom, a značajna razlika izme|u grupa AKI-2 i AKI-1 (p = 0,006) bila je izmerena tek drugi dan posle operacije. Upotreba stabla odlučivanja za klasifikaciju pacijenata sa manjim i sa ve}im rizikom od razvijanja oštećenja bubrega pokazala je da je Klotho u prvim satima nakon operacije bolji od kreatinina u diferencijaciji između pacijenata sa manjim i pacijenata sa većim rizikom od oštećenja bubrega. Predviđanje razvoja težeg oštećenja bubrega bilo je još bolje sa dodatnim uključivanjem ranih rezultata kreatinina u model. Zaključak: Serumski Klotho može biti koristan za diferencijaciju izme|u pacijenata sa manjim i pacijenata sa većim rizikom od razvijanja težeg oštećenja bubrega posle operacije srca koristeći CPB već u prvim satima nakon operacije

Validity of Klotho, CYR61 and YKL-40 as ideal predictive biomarkers for acute kidney injury: review study

ABSTRACT CONTEXT AND OBJECTIVE: Acute kidney injury (AKI) is still a headache for clinicians and scientists as a possible reason for increased death among intensive care unit (ICU) patients after invasive cardiac surgery. Furthermore, the diagnostic process for AKI using conventional biomarkers is not sufficient to ensure early warning of this condition because of the morbid influence of non-renal factors that definitively delay the time for the prognosis. These imposed limitations have led to significant amounts of research targeted towards identifying novel biomarkers for AKI with a sustained degree of sensitivity and specificity. Here, we reviewed previous studies conducted on the Klotho, CYR61 and YKL-40 biomarkers in relation to AKI. DESIGN AND SETTING: Review of the literature conducted in the Institute of Clinical Chemistry & Biochemistry, Ljubljana University Medical Center, Slovenia. METHODS: The literature was searched in PubMed and the Cochrane Library. From the database of this specialty, we selected 17 references that matched our context for detailed analysis and further investigation. RESULTS: The studies reviewed showed notable differences in their results relating to the diagnostic impact of Klotho, CYR61 and YKL-40 on early prediction of AKI. CONCLUSIONS: The results regarding the Klotho, CYR61 and YKL-40 biomarkers showed markedly equivocal performance in the previous studies and did not fulfill the expectations that these factors would form valid possible biomarkers for AKI

Nutrient supply systems and their effect on the performance of the Nile Tilapia (Oreochromis niloticus) and Lettuce (Lactuca sativa) plant integration system

Abstract The main aim of this work is to study the effect of different nutrient supply systems and their effect on the performance of the Nile Tilapia (Oreochromis niloticus) and Lettuce (Lactuca sativa var. crispa) plant integration system. To achieve that, five treatments having different culture systems (T1: Aquaculture (control), T2: Hydroponics (standard requirement: N = 210, P = 31, K = 234, Mg = 48, Ca = 200, S = 64, Fe = 14, Mn = 0.5, Zn = 0.05, B = 0.5, Cu = 0.02 and Mo = 0.01 ppm), T3: Aquaponics without nutrients addition, T4: Aquaponics with supplementary nutrients (KNO3, 101 g L−1, KH2PO4, 136 g L−1, Ca(NO3)2, 236 g L−1, MgSO4, 246 g L−1, K2SO4, 115 g L−1 and chelates for trace elements) in water (EC is 800 ppm) and T5: Aquaponics with supplementary nutrients spray on plants) were carried out. The previous systems were operated at three flow rates, namely, 1.0, 1.5 and 2.0 L h−1 plant−1. The various water quality parameters, plant growth and fish growth were studied. The result indicated that the highest values of N, P, k, Ca and Mg consumption rate were found with T2 and 1.5 L h−1 plant−1 of flow rate. The root length, fresh and dry of shoot and root for lettuce plants grown in T2 system was better than those grown in different culture system (T3, T4 and T5). Different culture systems showed significant effect on fish growth in terms of weight gain, specific growth rate and feed efficiency ratio. Higher growth rate was observed in treatment T3 as compared to other treatments. The production costs ranged from 2820.5 to 4885.4 LE ($ = 30.92 LE) for all culture systems

FN-Identify: Novel Restriction Enzymes-Based Method for Bacterial Identification in Absence of Genome Sequencing

Sequencing and restriction analysis of genes like 16S rRNA and HSP60 are intensively used for molecular identification in the microbial communities. With aid of the rapid progress in bioinformatics, genome sequencing became the method of choice for bacterial identification. However, the genome sequencing technology is still out of reach in the developing countries. In this paper, we propose FN-Identify, a sequencing-free method for bacterial identification. FN-Identify exploits the gene sequences data available in GenBank and other databases and the two algorithms that we developed, CreateScheme and GeneIdentify, to create a restriction enzyme-based identification scheme. FN-Identify was tested using three different and diverse bacterial populations (members of Lactobacillus, Pseudomonas, and Mycobacterium groups) in an in silico analysis using restriction enzymes and sequences of 16S rRNA gene. The analysis of the restriction maps of the members of three groups using the fragment numbers information only or along with fragments sizes successfully identified all of the members of the three groups using a minimum of four and maximum of eight restriction enzymes. Our results demonstrate the utility and accuracy of FN-Identify method and its two algorithms as an alternative method that uses the standard microbiology laboratories techniques when the genome sequencing is not available.Peer Reviewe