Jaffe Reaction Research Articles

#1413 High performance of ACLARA calculator of rat creatinine clearance for the chronic follow up of renal function

Abstract Background and Aims A neural network-based calculator to estimate rat creatinine clearance (ClCr), as a measurement of the glomerular filtration rate (GFR), from paired plasma creatinine concentration and body weight data has been recently developed [1]. Like human GFR estimating formulae, ACLARA averts long experimental procedures and reduces animal stress (by obviating the use of individual isolation in metabolic cages). This tool is publicly available for free use among the scientific community (https://idal.uv.es/aclara/). When collated with experimentally measured ClCr, ACLARA performs very well at comparing experiments in the whole, showing almost identical behavior of all experimental groups and conditions. Yet, ACLARA was trained mostly with data from short experiments involving acute changes in renal function, and from young rats. Fewer data from older animals was included in the development of the calculator. Thus, its field performance on longer evolution of renal function in adult rats has not yet been sufficiently validated. Accordingly, the aim of this work was to deepen in the assessment of ACLARA in the long-term evolution of rat ClCr. Method The evolution of measured (mClCr) versus ACLARA-estimated ClCr (eClCr) was compared at different time points over a period of 3 months in 20 male Wistar rats with chronic renal damage and 8 age-matched controls. In total, 176 data pairs of mClCr and eClCr were obtained. On the one hand, mClCr was determined by the usual experimental procedure [2]. Briefly, rats were allocated in metabolic cages to obtain 24-hour urine samples and urine flow (UF). Creatinine concentration was measured in the urine (Cru) and plasma (Crp) by a colorimetric assay based on the Jaffe's reaction, and mCrCl was then calculated as Cru x UF / Crp. On the other hand, eClCr was estimated with the ACLARA calculator from the corresponding Crp and body weight data. A Pearson's correlation study between mClCr and eClCr was carried out. Results Our results reveal that the information provided by the 90-day evolution of mClCr and eClCr is virtually identical at the whole experiment level (see Fig. 1). Furthermore, ACLARA seems to provide slightly more coherent and homogeneous information, as the considerable experimental error introduced by urine collection is averted in eClCr but present in mClCr. At the individual data level, ACLARA-estimated values are slightly lower than the corresponding measurements. The correlation between mClCr and eClCr was highly significant (r Pearson= 0.83; p < 0.0001). Conclusion ACLARA shows high performance at estimating CrCl in rats of growing age. Our results support its use also in chronic studies in which renal function must be evaluated. Funding This research was funded by grants from Instituto de Salud Carlos III (ISCIII), Ministerio de Ciencia e Innovación (PI21/01226 and PI21/00548 co-funded by the European Union; and RICORS2040, RD21/0005/0004, co-funded by the European Union – NextGenerationEU, Mecanismo para la Recuperación y la Resiliencia (MRR)) and from Consejería de Educación, Junta de Castilla y León (IES160P20), co-funded by FEDER funds. Joana Mercado-Hernández is recipient of a predoctoral fellowship from the Junta de Castilla y Leon (Spain) and the European Social Fund from the European Commission.

#1962 Development of triple-whammy induced pre-renal AKI is highly dependent on the hydration status

Abstract Background and Aims The concomitant use of diuretics, non-steroidal anti-inflammatory drugs (NSAIDs), and angiotensin system inhibitors is known as Triple Whammy (TW) and significantly increases the risk of acute kidney injury (AKI) due to a reduction in the glomerular filtration rate. Maintenance of hydration contributes to correct kidney perfusion, and effective renal haemodynamic regulation allows the kidneys to cope with everyday stressors and challenges, such as TW therapy. Interference with these adaptive mechanisms frequently occurs due to extrinsic (environmental conditions, lifestyle, drugs) or intrinsic (comorbidities, ageing) factors and may lead to dehydration, a negative body liquid balance with a high impact on health. Identifying modifiable risk factors that pose individuals at high risk of AKI following TW therapy is key for the search and prescription of preventive strategies in a personalized manner. The aim of this work was to study the role of hydration status on the renal effect of TW. Method Two-month-old male Wistar rats were divided into 3 experimental groups: Blood and 24-hour urine samples were collected at basal state (B), two days after water deprivation (D0), and two days after TW therapy (D2). Body weight was monitored during the experiment. Haematocrit was assessed from blood samples, and osmolality, concentration of creatinine (Jaffe reaction), and urea (Jung method) were determined in plasma. Urinary flow was recorded, and specific gravity, osmolality and creatinine concentration were measured in urine samples. Results Dehydration was observed after 48 h water deprivation, evidenced by a 12% weight loss and significant increases in haematocrit and plasma osmolality. As expected, during this dehydrated state rats excreted a very reduced volume of a highly concentrated urine with elevated levels of specific gravity and osmolality. No changes in renal function were observed after 48 h water deprivation, nor after TW treatment in fully hydrated rats. Administration of TW triggered significant alterations of the renal function, indicative of AKI, only in rats previously deprived of water for 48 h. These alterations were markedly increased when dehydration was partially sustained during the TW treatment by a 60% water restriction. Conclusion TW-induced AKI in young Wistar rats is facilitated when the hydration status worsens. In this sense, assessing hydration and implementing (re)hydration strategies may help to reduce or prevent TW-induced AKI. This study was supported by Project PI21/01226, funded by Instituto de Salud Carlos III (ISCIII) and co-funded by the European Union, and a grant from the Conserjería de Educación, Junta de Castilla y León (IES160P20), Spain, co-funded by FEDER funds. Noelia Diaz-Morales is recipient of a Juan de la Cierva-Formación postdoctoral contract (FJC2020-043205-I) funded by MCIN/AEI/10.13039/501100011033 and European Union “NextGenerationEU/PRTR”.

Dopamine infusion at typical infusion rates does not cause interference on plasma creatinine assays

a) ObjectivesDopamine is known to cause negative interference on enzymatic creatinine measurement. However, its effect on the Jaffe reaction, and its concentration required to interfere with enzymatic reactions, remain uncertain. This study was designed to study the interference of stable dopamine infusion on Jaffe and enzymatic creatinine assays, as well as the effect of dopamine infusion drip arm contamination on both creatinine assays. b) Design and MethodsFor the first part of the study, dopamine was spiked into pooled plasma samples at different concentrations to mimic the scenario of patients on dopamine infusion at an infusion rate between 2 and 20 μg/kg/min. For the second part, dopamine preparation of 2 g/L (same as the preparation used clinically) was mixed with pooled plasma samples at different proportions to mimic drip arm contamination. Creatinine concentrations were measured using Jaffe and enzymatic reactions. c) ResultsThe first part showed that creatinine measurements were not interfered by dopamine infusion at an infusion rate between 2 and 20 μg/kg/min. The second part showed that dopamine could negatively interfere with enzymatic creatinine assays, even with minute drip arm contamination. The effect on the Jaffe reaction was less significant. d) DiscussionCreatinine concentration could be reliably measured by Jaffe or enzymatic reactions if samples are from venous access sites other than the site of dopamine infusion. When dopamine interference on enzymatic creatinine assays is suspected, using the Jaffe reaction to cross-check may provide additional useful information.

Boron nitride nanotube-based creatinine biosensors for athlete physical condition monitoring

Monitoring creatinine levels is critical for evaluating renal function and muscular health in athletes. Creatinine is a waste product of creatine metabolism in skeletal muscle, and is filtered by the kidneys at a relatively constant rate. Elevated serum creatinine can indicate impaired kidney function, rhabdomyolysis, or skeletal muscle trauma. In athletes, creatinine levels increase after intense exertion due to muscle breakdown and myoglobin release. Abnormal creatinine signifies poor training adaptation, overtraining, or trauma. Therefore, close monitoring of creatinine in athletes provides insight into their physical condition. Current laboratory methods for measuring creatinine like Jaffe reaction and high-performance liquid chromatography are accurate but time-consuming, requiring blood sampling. Biosensors offer rapid, painless creatinine detection from small volumes, but current versions suffer from instability, use of mediators, and lack portability. Boron nitride nanotubes (BNNTs) are promising materials for electrochemical biosensing due to their structure, surface chemistry, and biocompatibility. Here, we report BNNT-based electrochemical biosensors for detecting creatinine in blood samples from athletes. BNNTs synthesized by induction thermal plasma technique were used to immobilize creatinine metabolizing enzymes. Creatinine levels were measured by chronoamperometry and compared to isotope dilution mass spectrometry. We demonstrate high sensitivity and selectivity of the BNNT biosensor with facile fabrication method and portable platform, promising for rapid creatinine monitoring in athletes. This can provide insight into muscular damage and renal function during training and competition.

Quantification of creatinine in whole blood by a paper-based device using an RGB sensor.

Creatinine, an important biomarker for renal functions, is often conventionally estimated using the gold standard Jaffe reaction from blood, which involves the usage of a spectrophotometric measurement, thus restricting its utilization scope in point-of-care settings. Here, we report the development of a method for the single-step quantification of creatinine from whole blood using a paper-based microfluidic device. Our platform uses Whatman filter paper integrated with an LF1 membrane. The on-chip separation of blood plasma is achieved through the LF1 membrane, while the Whatman component of the device contains the embedded reagents for the Jaffe reaction. The combination of two different grades of paper enables a single-step quantification of creatinine as the separated blood plasma traverses to the reaction zone through capillary imbibition. Colorimetric readouts were quantified using an RGB sensor instead of a smartphone, which is highly platform dependent and incurs a relatively higher cost compared to the other components in typical point-of care (POC) devices. Our sensor was integrated within a 3D box, thereby making the detection virtually instrument free and perfectly suited for POC settings. The limit of detection (LOD) of our device was 0.219 mg dL-1, which falls within the lower range of physiological values. The coefficient of determination (R2) for the linearity and median accuracy were 0.978 and 94.047%, respectively. The relative standard deviation (RSD) for precision measurements remained below 5% for the developed protocol. Furthermore, we validated the performance of our device with 35 clinical samples in laboratory settings against the gold standard measurements. Our Bland-Altman plot as well as t-test and chi-square test results clearly confirmed the validity of our device within a 95% confidence interval.

P-436 Non-Essential Trace Elements in women’s biofluids are associated with worse IVF outcomes in euploid single embryo transfer cycles.

Abstract Study question What is the non-essential trace element exposure impact on the reproductive outcomes of women undergoing IVF treatment? Summary answer Increased blood and urinary non-essential trace element concentration were associated with worse IVF outcomes. What is known already Lower fertility rates have been observed in recent decades in industrialized regions, increasing the need for assisted reproductive techniques. Factors responsible for this decline in human fertility include exposure to non-essential trace elements. Since these elements have no known biological functions, they are considered detrimental to the normal function of the organism. The most common elements evaluated so far have been those traditionally considered toxic, such as heavy metals and metalloids (mercury, lead, cadmium and arsenic), for which inconsistent negative associations with different IVF treatment variables have been described. Study design, size, duration 51 women who underwent an euploid single-embryo transfer (SET) after PGT-A analysis, were included. Nine non-essential elements (barium (Ba), strontium (Sr), rubidum (Rb), arsenic (As), tin (Sn), cesium (Cs), mercury (Hg), lead (Pb), antimonium (Sb)) were measured in follicular fluid (FF), whole blood (B) and urine (Urine-VOR), all collected at vaginal oocyte retrieval day (VOR), and urine collected at transfer day (Urine-T). These measurements were correlated with IVF clinical outcomes. Participants/materials, setting, methods Quantification of non-essential elements in the four biofluids were performed by inductively coupled plasma mass spectrometry (ICP-MS). Urine concentrations were normalized by creatinine, quantified by Jaffe reaction. Generalized linear models were employed to explore ovarian response variables and embryological IVF outcomes (associations, estimated as percentile 20th to 80th increase (95% confidence intervals (CI)). Associations with IVF clinical outcomes were estimated by odds ratio (95%CI). Both unadjusted and age-BMI-race-smoking-adjusted models were applied. Main results and the role of chance Participants had a median age of 39 years [Inter Quartile Range (IQR): 31.37, 36.50] and BMI of 22.97 kg/m2 [IQR: 20.63, 25.12], and 50% had never smoked. In adjusted models, significantly negative associations were found between blood Hg concentration and relative frequency of fertilized embryos [0.73 (0.56,0.96), p = 0.024], blastocyst arrival [0.64 (0.44,0.93), p = 0.019], and euploid embryos [0.60 (0.37,0.98), p = 0.044]. We also found negative associations for urine-VOR Sn and blastocyst arrival [0.72 (0.53,0.98) p = 0.038] and euploid embryos [0.55 (0.39,0.77), p < 0.001]. In the case of urine-T, higher concentrations of St were associated with lower number of retrieved oocytes [0.71 (0.57,0.90), p = 0.006], relative frequency of mature oocytes [0.75 (0.62,0.90), p = 0.003], fertilized embryos [0.72 (0.62,0.83), p < 0.001], blastocyst arrival [0.68 (0.56,0.84), p < 0.001] and euploid embryos [0.83 (0.69,1.00), p = 0.048]. urine-T Cs was associated with lower proportion of mature oocytes retrieved [0.65 (0.42,0.99), p = 0.046] while both Rb and Cs was associated with lower proportion of euploid embryos [0.066 (0.44,0.99), p = 0.048; 0.55 (0.34,0.89), p = 0.019, respectively]. Regarding IVF clinical outcomes, in fully adjusted models, our results suggest that higher concentrations of As in urine-T were associated with lower probability of live birth [0.03 (0.00,0.36); p = 0.038] and reproductive goal (live birth/all participants) [0.03 (0.00,0.36); p = 0.038]. Limitations, reasons for caution Further studies are needed to confirm this association in greater populations, including the measurement of different element species. Wider implications of the findings Exposure and circulating levels of these non-essential elements have a significant impact on IVF outcomes. These data highlight the need to further study non-essential trace elements to identify and characterize how may be affecting IVF. Trial registration number not applicable

#4248 COMBINED EFFECT OF DEHYDRATION, HYPERTENSION, AND AGE ON RENAL FRAILTY

Abstract Background and Aims Renal frailty (RF) is a premorbid and, at least partly, modifiable condition arising from diminished renal functional reserve and defective adaptive response capacity predisposing to acute kidney injury (AKI). RF ensues from subclinical wear or distortion of the renal haemodynamic and tubular homeostatic responses that defend the renal excretory function from supervening circumstances (i.e., stressors such as drugs). In this work, we aimed to study the impact of RF generated by the combined effect of the hydration state, hypertension and ageing on the nephrotoxicity developed by a low dose of cisplatin in the rat. Method Young Wistar rats, young spontaneously hypertensive rats (SHR), and aged Wistar rats were used. Rats in each group were randomly subject to four experimental conditions: - Control: rats with ad libitum water intake, receiving vehicle (0.9% NaCl, i.p). - Cisplatin: rats with ad libitum water intake, treated with 2.5 mg/kg, i.p. cisplatin. - Water deprivation: rats deprived of water for 48 h, receiving vehicle (0.9% NaCl, i.p.). - Water deprivation + cisplatin: rats deprived of water for the 48 h prior to 2.5 mg/kg i.p. cisplatin administration. Tail vein blood and 24-hour urine samples were collected at basal time (B), after water deprivation (D0), and 4 days after cisplatin/vehicle administration (D4, day of maximum kidney damage). Haematocrit was measured and plasma osmolality, plasma creatinine (Jaffe reaction) and urea (Jung method) concentrations were determined. Urine samples were analysed for volume (urinary flow), osmolality, and proteinuria (Bradford method). Body fluid composition was measured by bioimpedance spectroscopy with an ImpediVETTM VetBIS1 device. Results All rats showed clear signs of dehydration after 48 hours of water deprivation: weight loss, increases in plasma osmolality and haematocrit (more pronouncedly in young Wistar and SHR than in aged animals), and reduced flow of a highly concentrated urine (i.e., with elevated osmolality). Bioimpedance analysis revealed a parallel loss of intracellular and extracellular water and net loss of total body water (explaining most of weight loss), with no changes in the liquid distribution between the intracellular and extracellular compartments. Dehydration was reverted after 4 days of rehydration in vehicle-treated rats, and no alteration of renal function was observed due to water deprivation. Administration of cisplatin in young, normohydrated Wistar rats showed minimal loss of renal function that was not worsened by water deprivation. Hypertensive rats showed completely normal renal function when cisplatin was administered under normohydration. Interestingly, however, dehydrated SHR did suffer a cisplatin-induced AKI (significant increases in plasma creatinine, plasma urea and proteinuria). In aged Wistar rats, cisplatin compromised renal function (as supported by increases in plasma creatinine and urea concentration), and this effect was significantly amplified by water deprivation. Conclusion This study suggests that dehydration in fully competent young rats is not in itself a significant risk factor for AKI, while the combination of dehydration and hypertension in young rats significantly increases the risk of AKI. Furthermore, ageing poses animals in a state of frailty that renders their kidneys unable to respond to stressors (such as a low dose of cisplatin). Frailty is boldly worsened in dehydrated aged animals. In perspective, models reproducing conditions of increased vulnerability to AKI provide useful tools to search for biomarkers pre-emptively predicting undesired health outcomes before exposure to stressors, and for developing preventing strategies. This study was supported by Project PI21/01226, funded by Instituto de Salud Carlos III (ISCIII) and co-funded by the European Union, and a grant from the Consejería de Educación, Junta de Castilla y León (IES160P20), Spain, co-funded by FEDER funds. Noelia Diaz-Morales is recipient of a Juan de la Cierva-Formación postdoctoral contract (FJC2020-043205-I) funded by MCIN/AEI/10.13039/501100011033 and European Union “NextGenerationEU/PRTR”.

Evaluasi Kadar Kreatinin dan Mikroalbumin dengan HbA1C <6% pada Pasien Diabetes Mellitus di Samarinda

Diabetes Mellitus (DM) is a metabolic disease caused by high blood glucose levels (hyperglycemia). Hyperglycemia is caused due to lack of insulin secretion in the body. Glycemic control in diabetic patients is seen in HbA1C levels. One of the complications of DM is diabetic nephropathy. Renal function abnormalities associated with diabetic nephropathy are indicated by increased levels of creatinine and microalbumin. This shows that it is necessary to monitor kidney function in DM. Monitoring DM patients with HbA1C levels <6.5% is desirable, with these levels indicating that glycemic control is good. This study aimed to determine the relationship between creatinine and microalbumin levels with HbA1C <6.5% in Diabetes Mellitus patients in Samarinda. This type of research is analytic with a cross-sectional design. Spearman correlation statistical test. The sample of this study was 107 DM patients with HbA1C levels <6.5% who did medical checkups at the Media Farma Clinic, Samarinda. Sampling is done by Total Sampling. HbA1C examination method with turbidimetry. Method of checking creatinine by Jaffe reaction. Method of examination of microalbumin with immunoturbidimetry. All parameters are quality controlled before measurement. The measurement results were validated by a clinical pathologist. The results showed that the correlation of creatinine with HbA1C <6.5% obtained p value = (0.826), microalbumin with HbA1C <6.5% obtained p value = (0.146), and creatinine with microalbumin obtained p value = (0.082). All these results indicate that H0 is accepted and Ha is rejected. This study concludes no significant relationship exists between creatinine and microalbumin values with HbA1C levels <6.5% in Diabetes Mellitus in Samarinda, especially Media Farma Clinic.

Determination of creatinine in urine and blood serum human samples by CZE-UV using on-column internal standard injection.

Creatinine is a well-stablished biomarker for kidney malfunctions and for normalization parameter of urinary quantitative information. Recently, metabolic studies have been discovering other functionalities for creatinine tests in human urine and blood serum. In this work we present an enhanced capillary electrophoresis (CE) based protocol for determination of creatinine. CE is a high-throughput separation technique that have been getting attention through the last decades and might be considered to be adopted as an analytical instrumentation for clinical purposes. In the proposed method, we performed a short injection program with on-column addition of internal standard. Additionally, the method allows a simultaneous screening of non-proteinogenic amino acids that could be considered for metabolomics purposes. We design a pilot study that successfully estimated the creatinine value in 100 urine samples with (2.85±1.78) mg dL-1 LOD; (8.24±5.93) mg dL-1 LOQ and 82.4% accuracy. Considering that serum creatinine is also included in the clinical laboratory routines for estimated Glomerular Filtration Rate dosage, the method was complementary applied to 10 blood serum samples, which resulted in a model with (0.4±0.2)mgdL-1 LOD; (2.0±0.6)mgdL-1 LOQ and 83.8% of accuracy. All results were in agreement with reference values. The proposed method promotes a great analytical frequency and reproducibility with enhanced specificity compared with the ongoing protocol by Jaffe's reaction, thereby proving to be useful as an alternative for creatinine exams that might help complete a diagnosis of a series of health-related issues.

Relationship Between Estimated Glomerular Filtration Rate And Microalbuminura In Hiv/aids Patients: The Kano Experience

Background: Chronic Kidney Disease is one of the major complications that may occur in HIV/AIDS, usually manifesting as reduced glomerular filtration rate (GFR) and albuminuria. Microalbuminuria as an early marker of CKD may help in the early identification of those at risk of CKD in this cohort of patients, allowing for early institution of appropriate measures. The aim of the study was to determine the relationship between microalbuminuria and different levels of estimated glomerular filtration rate (eGFR) in HIV/AIDS adult patients in Aminu Kano Teaching Hospital, Kano, Nigeria. Methods: A descriptive cross-sectional study was conducted among 500 HIV/AIDS participants. Basic biodata was documented. Blood samples for serum creatinine and urine samples for urinary albumin and creatinine measurements were collected. Urinary and serum creatinine levels were determined using the Jaffe reaction, and urine albumin was measured using the turbidimetric immunoassay. The eGFR was calculated using Cockcroft and Gault formular and results were interpreted using Kidney Disease Improving Global Outcome (KDIGO) criteria. Comparison of categorical variables for associations was conducted using Fisher exact test. Pearson correlation was done to determine the strength of the relationship between eGFR and microalbuminuria. Results: Estimated Glomerular filtration <60ml/min/1.73m2 showed a statistically significant association with microalbuminuria on bivariate analysis but was found not to be an independent predictor of microalbuminuria with, at best, a weak negative correlation between the two variables. Conclusion: eGFR is not an independent predictor of microalbuminuria in HIV/AIDS patients.

Status of some biochemical markers in Sprague dawley rats following sub-chronic consumption of alum- treated water

Alum is a double salt of aluminum and potassium sulfates. The aim of this study was to investigate the status of some biochemical markers in Sprague-dawley rats following sub-chronic consumption of alum- treated water. It consisted of 21 rats of 3–4 months old weighing roughly 122 ± 0.5 g. They were divided into three groups at random with 7 rats / group. The control group received 15 ml of distilled water daily for 30 days, while experimental groups one and two received 15ml of 0.04% concentration of alumtreated water for 15 days daily and 30 days respectively. Following this, 5 ml of blood specimen was withdrawn via cardiac puncture and introduced into corresponding lithium heparin anti-coagulated bottles. These specimens were spun at 1,500 revolutions per minute for 10 minutes using an 800 D macro centrifuge to extract the plasma that were utilized for the measurement of alanine aminotransferase, aspartate aminotransferase (colorimetric method), urea (urease Berthelot method), creatinine (Jaffe reaction method) and C-reactive protein (Latex turbidimetry method). Using SPSS 23.0 to perform statistical analysis on the study's findings, it was discovered that there were no significant differences between the experimental group one and the control group for all the measured biochemical parameters (p>0.05). However, in comparison to the control group, the results of the experimental group two revealed significant elevations (p<0.05) in revealed significant elevations (p<0.05) in plasma levels of alanine aminotransferase, aspartate aminotransferase and C-reactive protein. It is concluded that the plasma concentrations of alanine aminotransferase, aspartate aminotransferase and C-reactive protein may be altered in Sprague-dawley rats following consumption of 15 ml of 0.04% concentration of alum- treated water for 30 days. Thus, it is recommended that this study in a well guided manner be carried out in humans to ascertain if the findings will be consistent.

Assessment of Electrolyte Imbalance and Associated Factors Among Adult Diabetic Patients Attending the University of Gondar Comprehensive Specialized Hospital, Ethiopia: A Comparative Cross-Sectional Study.

Electrolyte imbalance refers to altered electrolyte levels that predominantly affect diabetic patients due to hyperglycemia which rise plasma osmolality and impaired renal function, contributing to a change in electrolyte level. Therefore, this study aimed to assess the prevalence of electrolyte imbalance and its associated factors among diabetic patients and healthy control groups attending the University of Gondar Comprehensive Specialized Hospital. A comparative cross-sectional study was conducted on 130 diabetic patients and 130 diabetes-free controls. Sociodemographic, behavioral and clinical data were collected using a structured questionnaire. After measuring anthropometric parameters, 5mL of the blood sample was collected. Electrolytes were measured based on ion-selective electrode methods. Fasting blood glucose, and creatinine were measured by spectrophotometric enzyme hexokinase, and Jaffe reaction methods, respectively. The data was entered into Epi-data version 4.6 and analyzed using STATA version 14. Mann-Whitney U-tests and independent t-tests were used for comparison. Multiple logistic regression analysis was done to determine the factors associated with electrolyte imbalances. P-value <0.05 was considered statistically significant. The overall prevalence of electrolyte imbalance among diabetic patients and controls were 83.07% and 52.31%, respectively. The mean of Na+ and the median level of Mg2+ and Ca2+ were significantly decreased. However, the mean level of Cl- was significantly increased in diabetic patients as compared to control groups. In multivariable logistic regression: alcohol consumption AOR = 3.34 [1.02-10.9], no formal education AOR = 5.38 [1.14-25.4], hyperglycemia AOR = 6.32 [2.04-19.5], and urbanization AOR = 5.6 [1.44-22.3] showed significant association with electrolyte imbalance. Diabetic patients have more likely to develop electrolyte imbalance than control groups. Diabetic participants showed significantly reduced Na+, Mg2+, and Ca2+ levels and significantly increasing CI- levels when compared to control groups. Hyperglycemia, alcohol drinking habits, urbanization, and no-formal education were statistically significantly associated with electrolyte imbalance.

COMPARATIVE EVALUATION OF CREATININE IN SERUM AND SALIVA OF CHRONIC KIDNEY DISEASE PATIENTS

Background &amp; objectives: Chronic kidney disease (CKD) is progressive reduction in renal function. Renal excretory function can be assessed by measurement of creatinine concentrations in plasma which illustrates the ltration capacity of the glomerulus. The increased serum creatinine levels in CKD patients create a concentration gradient that facilitates increased diffusion of creatinine from serum into the saliva. Saliva has many advantages as a diagnostic media over serum, which includes non-invasive collection, good patient compliance, minimal risk of contracting infection, repeated sampling possible and is cost effective. The purpose of this study was to estimate and correlate serum and salivary creatinine in patients with CKD and to evaluate use of salivary creatinine as an alternative biomarker to serum creatinine in CKD patients. A total sample of 40 subjects aged 25 -6 Methods: 5yrs, inclusive of both genders comprising 20 cases of healthy controls (Group I) and 20 diagnosed cases of CKD (Group II) were selected. 2ml of blood and unstimulated whole saliva was collected from both the groups. The blinded samples were processed in autoanalyzer using Jaffe's kinetic reaction. The collected data were subjected to Statistical analysis. In the present Results: study we found signicant correlation of serum creatinine and salivary creatinine in both controls and CKD. Serum creatinine (6.21 ±4.02) and Salivary creatinine (0.70± 0.61) were both increased in CKD cases compared to Controls Serum creatinine (0.83±0.21) and Salivary creatinine (0.11± 0.07). Serum creatinine and salivary creatinine were xii both increased as CKD progressed. A sensitivity of 75% and specicity of 90% of salivary creatinine was found. We therefore recommend Conclusion: salivary creatinine as a biomarker in CKD and also as an alternative biomarker to serum creatinine in detecting and staging CKD, for assessing disease progression, therapeutic modalities and in screening of large population

OVERVIEW OF SERUM CREATININE LEVELS IN TRADITIONAL GOLD MINING WORKERS IN EAST MAMUNGA’A VILLAGE

Creatinine is a kind of chemical in the body indicating damage to body cells. The levels in the blood indicate how much damage to body cells. The purpose of this study was to determine the description of serum creatinine levels in traditional gold mining workers in East Mamunga'a Village in 2021.This research is descriptive with a quantitative approach. The population in this study were all traditional gold mining workers with a total sample of 35 people. The sampling method used Accidental Sampling. The examination of serum creatinine levels using the Jaffe Reaction method and measured using a Spectrophotometer biosystem BTS 350. The types of data used were primary data and secondary data with data sourced from interviews and documentation. Furthermore, the data were analyzed univariate.The results show that 15 samples (42.9%) of serum creatinine are abnormal and 20 samples (57.1%) are normal. Serum creatinine levels that do not increase or are declared normal are caused by traditional gold mining workers adopting a healthy lifestyle. The conclusion is that traditional gold mining workers experience an increase in serum creatinine levels less than normal. The need to maintain a healthy lifestyle such as adequate rest during mining activities, consuming mineral water, and not consuming excessive supplements so as not to increase serum creatinine levels.

A narrative review on laboratory investigations of serum creatinine and solutions to problems therein

Introduction: It is of utmost importance to assess renal function accurately for the proper management of renal failure. Serum creatinine level is used to calculate the creatinine clearance which is used to measure the glomerular filtration rate (GFR) and in grading different stages of chronic kidney disease (CKD). Therefore, the accurate measurement of serum creatinine is of great importance. This review aimed to explain the different methods used in serum creatinine determination and solutions to problems in some of the methods. Methods: A narrative review was conducted with 28 articles searched through the Web searching engine Google Scholar and academic research databases using keywords creatinine, isotope dilution mass spectrometry, high performance liquid chromatography (HPLC), Jaffe reaction, creatinase enzyme and bilirubin. Results: There are different methods available in the determination of serum creatinine level in the medical laboratory sector including isotope dilution mass spectrometry, HPLC, Jaffe reaction and creatinase enzymatic method. Conclusion: Jaffe reaction is the most widely used method to determine serum creatinine levels worldwide. However, this method has several drawbacks and corrective methods are adopted.&#x0D; &#x0D; Keywords: Creatinine, Isotope dilution mass spectrometry, High Performance Liquid Chromatography, Jaffe reaction, Creatinase method, Bilirubin

A smartphone-integrated portable rotating platform for estimation of concentration level of plasma-creatinine using whole human blood

The measurement of creatinine concentration is performed to monitor the renal health. The devices available in modern clinical laboratories for measuring creatinine concentration are accurate and provide results rapidly but may be prohibitively expensive for resource-poor settings. Therefore, developing an inexpensive yet accurate device for measuring creatinine concentration is needed. Consequently, we developed a simple, affordable, and portable spinning disc for measuring plasma-creatinine concentration with 10 μL of whole human blood.5 μL of the alkaline picrate solution is loaded into the device and rotated at 1000 rpm to transport this solution to the periphery of the microchannel. Further, 10 μL whole blood is loaded in the same channel and spun at 1300 rpm for 10 min. The creatinine in plasma reacts with alkaline picrate (Jaffe reaction), and the color of the mixture changes to yellow-orange color. The resulting color is captured with a smartphone, and creatinine concentration is estimated using an in-house developed app (CREA-SESE).The value of creatinine measured with the present device and the gold standard device are highly correlated (R2 = 0.998). The bias and standard deviation of the difference between the two measurements are 0.134 mg/dL and 0.143 mg/dL. This study demonstrates the feasibility of a simple, inexpensive, and portable rotating device for measuring creatinine concentration using 10 μL of whole human blood, which can easily be deployed to the underserved population in resource-constrained settings to monitor renal diseases.

Role of predictable biomarkers in early detection of cardiovascular events in Chronic Kidney Disease III and IV

Abstract This comes about because of a lack of predicted biomarkers in the risk analysis of CVD events in chronic kidney disease (CKD) patients. The present study aimed to determine the clinical utility of independent, predictable biomarkers such as serum creatinine, estimated Glomerular Filtration Rate (eGFR), high sensitive C-Reactive protein (hsCRP), fibrinogen and lipid profile as early predictors of CVD in CKD at stage III/IV. Methods. This is a case-control study that includes a sample size of 100 patients of cases and 100 patients of controls who were recruited from November 2020 to April 2021, from the Nephrology department of the Visakhapatnam tertiary care teaching hospital, and present with chronic kidney disease – stage III/IV. The subjects’ general conditions (age, gender, height, weight, systolic blood pressure, diastolic blood pressure, and smoking history); underlying diseases (coronary heart disease and diabetes mellitus) were recorded. Fasting venous blood samples were collected under aseptic conditions from the study group after taking informed consent. The measurement of serum creatinine was performed by modification of kinetic Jaffe reaction. The Cockcroft-Gault equation was used to calculate eGFR in both cases and controls. CRP testing was done with a Cobas C311 analyzer, using immunoturbidimetric assay. The Fibroquant kit from Tulip was employed to measure fibrinogen levels in blood samples, and enzymatic methods were applied for lipid profile analysis. Results. In this study, higher mean values of hsCRP (34.28 mg/dl), increased serum creatinine levels (2.876 mg/dl), reduced eGFR (28.37 mls/min), high levels of serum fibrinogen (291.6 mg/dl), and cholesterol (214.5 mg/dl), HDL (28.34 mg/dl), TG (162.1 mg/dl), VLDL (32.41 mg/dl) and LDL (153.77 mg/dl) were found to be independent predictors of assessment of CV events in patients with CKD stages III and IV as determined by Chi-square test. Conclusion. A prompt and accurate assessment of cardiovascular risk in CKD patients would enable more aggressive and focused treatment of the individuals who are most in need of preventive interventions to decrease incident rates.

P-670 Phytoestrogen concentration in women’s urine and follicular fluid are associated with better IVF outcomes in euploid single embryo transfer cycles

Abstract Study question What is phytoestrogen exposure impact on the reproductive outcomes of women undergoing IVF treatment? Summary answer Increased urinary and follicular fluid phytoestrogen concentration were associated with better IVF outcomes. What is known already The impact and safety of phytoestrogens, plant-derived isoflavones with estrogenic activity predominantly present in soy, on female reproductive health and on IVF outcomes continues to be hotly debated. Numerous studies suggest healthy benefits of phytoestrogen consumption: protection against reprotoxic substances, reductions in breast and other cancers, fibroids, menopausal symptoms, cardiovascular disease, inflammation, and metabolic syndrome and obesity. However, many studies suggest a harmful effect due to the endocrine-disruptor properties of phytoestrogens in both male and female reproductive tissues. Its impact on IVF treatments success is still understudied. Study design, size, duration 60 women attending IVI-RMA New Jersey undergoing IVF with single-embryo transfer (SET) of euploid good-quality blastocyst after PGT analysis were recruited. Patients underwent PGT-SET cycles following standard protocols. Concentrations of two phytoestrogens (daidzein and genistein) were measured in follicular fluid (FF) and urine collected at oocyte pick-up (UOP) and urine collected at transfer day (UT). These measurements were correlated with IVF clinical outcomes. Participants/materials, setting, methods Participants were aged 18-42 years old 18.5-29.9 kg/m2 BMI. Measure of both phytoestrogens’ concentration in follicular fluid and urine samples collected at two different time-points was performed by UPLC-MSMS, with internal standards. Urine phytoestrogen concentrations were normalized by creatinine, measured by Jaffe reaction. Generalized linear models were employed to explore IVF outcomes association, estimated as percentile 20th to 80th increase (95% confidence intervals), among log-transformed phytoestrogen concentration. Both unadjusted and age-BMI-adjusted models were applied. Main results and the role of chance In fully adjusted models, higher sum of phytoestrogen concentration in follicular fluid (FF) were significantly associated with higher estradiol at hCG [1.31 (1.04, 1.65), p = 0.023]; whereas the sum of creatinine-corrected phytoestrogens measured in oocyte pick-up urine (UOP) were significantly associated with higher MII oocytes number [1.29 (1.07, 1.56), p = 0.008] and higher fertilized oocytes number [1.34 (1.10, 1.64), p = 0. 004]. Regarding clinical IVF outcomes, in fully adjusted models a higher follicular fluid and oocyte pick-up urine phytoestrogens concentration was significantly associated with higher implantation ratio [8.60 (1.09, 68.03), p = 0. 023] for FF, and [5.01 (0.96, 26.21), p = 0. 05] for UOP; higher clinical pregnancies ratio [5.58 (1.31, 23.73), p = 0.017] for FF, and [3.25 (0.99, 10.63), p = 0.045] for UOP; higher live newborns ratio [5.58 (1.31, 23.73), p = 0.017] for FF, and [3.25 (0.99, 10.63), p = 0.045] for UOP; and higher number of women with live newborns among those who started treatment [4.29 (1.28, 14.38), p = 0.016] for FF, and [2.83 (0.99, 8.07), p = 0.047] for UOP. Limitations, reasons for caution Further studies are needed to confirm this association in other populations, including the measurement of other estrogenic isoflavones and its metabolites. Wider implications of the findings This is the first work evaluating the association between phytoestrogens concentrations in different biofluids and IVF reproductive outcomes. Our results, using an euploid SET, suggest a beneficial effect of phytoestrogen consumption in women undergoing IVF treatment, further studies are needed to assess the benefit of phytoestrogen supplementation on IVF outcomes. Trial registration number -

Construction of Jaffe reaction-based SERS chip for determination of trace picric acid

Picric acid (PA) has a severe environmental risk and security problem owing to its unstable property and high toxic. On-field selective and sensitive detection of trace PA is also a challenging task. PA molecules due to poor affinity to surface of noble metal nanoparticles are hardly detectable by surface-enhanced Raman scattering (SERS) method. In this work, silver shell and gold core nanoparticles with inositol hexaphosphate inner gap (Au@IP6@AgNPs) is prepared and then two-dimension (2D) substrate at the ITO surface (designated as ITO-Au@IP6@Ag) is formed via self-assembly. For capturing PA, we firstly modify creatinine (CRN) molecules onto the ITO-Au@IP6@Ag designated as ITO-Au@IP6@Ag@CRN to construct a specific SERS chip for PA, taking advantage of Jaffe reaction between PA and CRN. The generated Jaffe Complex has the greater Raman scattering section with respect to PA. Therefore, such chip-based SERS array for PA shows a good sensitivity and selectivity and the limit of detection at 3.9 × 10−11 mol/L could be reached. As real applications, the ITO-Au@IP6@Ag@CRN chips have been used for identification of PA in environmental samples such as clothing, envelope, and luggage, and determination of trace PA (~10−7 mol/L) in lake water. It demonstrates a promising perspective for on-site detection of the explosive for public security.

Glomerular filtration rate in children with juvenile idiopathic arthritis

Today, it is proved that kidney injury in children with juvenile idiopathic arthritis (JIA) is characterized by a subclinical course and most often remains undiagnosed. One of the main methods to diagnose kidney lesions is determination of estimated glomerular filtration rate (eGFR). The aim of the study was to determine the most optimal method for eGFR evaluation in children with JIA by comparing different calculation methods based on serum creatinine (original Schwartz formula, Counahan-Barratt formula), serum cystatin C (cystatin C-based equation and Hoek formula). Materials and methods. Eighty children with JIA were examined. Serum creatinine level was determined twice (first and third month of study) by the colorimetric kinetic Jaffe reaction, serum cystatin C concentration was evaluated once, in the third month of the study by enzyme immunoassay. We used methods of variation statistics. Results. Parameters of eGFR by cystatin C-based equation 2012 compared to the original Schwartz formula and Counahan-Barratt formula amplified the rate of eGFR disorders by 81.3 % (p &lt; 0.001) and 47.3 % (p &lt; 0.001) in the first month of the study and by 81.3 % (p &lt; 0.001) and 55.0 % (p &lt; 0.001) in the third month, respectively. A similar comparison of the results of Hoek formula with Schwartz and Counahan-Barratt formula showed that according to Schwartz formula, a decrease in the GFR was 41.3 % less frequent (p &lt; 0.001) in both the first and third month. Counahan-Barratt formula in the first study of serum creatinine showed a difference in eGFR compared to Hoek formula by 7.5 % (p &lt; 0.05). During the second study of serum creatinine, the frequency of eGFR reduction according to Counahan-Barratt formula was detected by 15.0 % less often than according to Hoek method (p &lt; 0.09). Conclusions. Changes in renal function differ depending on the formula of eGFR evaluation. The most optimal method to determine eGFR in children with JIA is Hoek formula based on serum cystatin C. In case of limited ability to analyze serum cystatin C, Counahan-Barratt formula based on serum creatinine can be used.