Urine Of Diabetic Patients Research Articles

Utilizing machine learning algorithms for precise discrimination of glycosuria in fluorescence spectroscopic data

Fluorescence spectroscopy coupled with a random forest machine learning algorithm offers a promising non-invasive approach for diagnosing glycosuria, a condition characterized by excess sugar in the urine of diabetic patients. This study investigated the ability of this method to differentiate between diabetic and healthy control urine samples. Fluorescent spectra were captured from urine samples using a Xenon arc lamp emitting light within the 200 to 950 nm wavelength range, with consistent fluorescence emission observed at 450 nm under an excitation wavelength of 370 nm. Healthy control samples were also analyzed within the same spectral range for comparison. To distinguish spectral differences between healthy and infected samples, the random forest (RF) and K-Nearest Neighbors (KNN) machine learning algorithms have been employed. These algorithms automatically recognize spectral patterns associated with diabetes, enabling the prediction of unknown classifications based on established samples. Principal component analysis (PCA) was utilized for dimensionality reduction before feeding the data to RF and KNN for classification. The model’s classification performance was evaluated using 10-fold cross-validation, resulting in the proposed RF-based model achieving accuracy of 96 %, specificity of 100 %, sensitivity of 93 %, and precision of 100 %. These results suggest that the proposed method holds promise for a more convenient and potentially more accurate method for diagnosing glycosuria in diabetic patients.

Antibiogram Pattern of Multidrug Resistance of Gram-negative Extended Betalactamase Isolates from Urine of Diabetic Patients

Background: The rate of multidrug resistant bacteria is the most worrisome in the health setting because it often associates with nosocomial infection. Objectives: This present study aim at, evaluating the antibiotic pattern of multidrug resistance of Gram-negative extended betalactamase isolates from urine of diabetic patients. Methods: This is a cross sectional study carried out at Nnamdi Azikiwe University Teaching Hospital with a total of 110 diabetic patients and were recruited using convenient sample method. The patients were instructed on how to collect the mid-stream urine samples without contamination. The samples were cultured and characterized the isolates following standard bacteriological methods. The isolated bacteria were subjected to sensitivity test pattern using agar disc diffusion techniques. The multidrug resistant isolates were physically checked for the extended spectrum betalactamase production. Obtained data was subjected to these statistical tools; descriptive statistics, chi-square and pair-test using Statistical Package for the Social Science (SPSS version 26). Results: Out of 110 urine samples assessed for significant growth of bacteria, it was found that, 55(27%) were the Gram negative bacteria (GNB).The highest prevalence of isolates were E. coli 30(54.6%), followed by Klebsiella pneumonia 14 (25.5%), Pseudomonas aeruginosa 6 (10.9%), and Proteus species 5 (9.09%) respectively. The most occurrence of the MDR bacteria were found in Augmentin 46 (83.6%) as well as as Cefuroxime 43 (78.2%) respectively. The significant mean range of Augumentin and Cefixime resistant to ESBL producers were (0.53±1.2; p = 0.000) and (0.600 ± 0.974; p = 000) respectively. Conclusion: The study detected the high proportion of multi-drug resistant isolates and the most occurrence were found in E. coli and Klebsiella pneumoniae. Regular monitoring, conducting, supervising, or management of antibiotics and molecular biomarkers for drug resistance are paramount to curtail the rate of drug-resistant pathogens.

Urinary insulin signaling pathway related proteins may serve as potential biomarkers for monitoring diabetes mellitus without hypertension and hyperlipidemia.

The insulin signaling pathway plays an important role in the development of diabetes mellitus. The expression of insulin signaling pathway related proteins in the urine of diabetic patients has not been reported. The aim of this study was to analyze and verify the expression of insulin signaling pathway related proteins in the urine of diabetic patients without hypertension and hyperlipidemia, and to explore their clinical application value. Based on data-independent acquisition proteomics technology and bioinformatics, the urinary protein expression profile of diabetic patients without hypertension and hyperlipidemia was established. Western blot and enzyme-linked immunoassay were performed to verify the expression of insulin signaling pathway related proteins in the urine of diabetic patients. Sixteen proteins related to the insulin signaling pathway were screened in urine, and 7 of them were differentially expressed in the urine of diabetic patients without hypertension and hyperlipidemia. Further quantitative analysis showed that the downregulation of protein kinase CAMP-dependent type II regulatory subunit α, growth factor receptor bound protein 2, and guanine nucleotide-binding protein G(s) in the urine of diabetic patients without hyperlipidemia and hypertension was consistent with the preliminary screening results. In this exploratory study, we detected the expression of insulin signaling pathway related proteins in the urine of diabetic patients without hypertension and hyperlipidemia. protein kinase CAMP-dependent type II regulatory subunit α, growth factor receptor bound protein 2, and guanine nucleotide-binding protein G(s) in the urine of diabetic patients were downregulated, which was associated with diabetes. They may be promising noninvasive biomarkers for monitoring diabetes.

Pyruvate Kinase M2: A New Biomarker for the Early Detection of Diabetes-Induced Nephropathy.

Diabetic nephropathy (DN) is a common complication of diabetes. DN progresses to end-stage renal disease, which has a high mortality rate. Current research is focused on identifying non-invasive potential biomarkers in the early stage of DN. We previously indicated that pyruvate kinase M2 (PKM2) is excreted in the urine of rats after cisplatin-induced acute kidney injury (AKI). However, it has not been reported whether PKM2 can be used as a biomarker to diagnose DN. Therefore, we try to compare whether the protein PKM2 can be detected in the urine samples from diabetic patients as shown in the results of DN models. In this study, high-fat diet (HFD)-induced Zucker diabetic fatty (ZDF) rats were used for DN phenotyping. After 19 weeks of receiving a HFD, the DN model's blood glucose, blood urea nitrogen, and serum creatinine levels were significantly increased; severe tubular and glomerular damages were also noted. The following protein-based biomarkers were increased in the urine of these models: kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), and PKM2. PKM2 had the earliest detection rate. In the urine samples of patients, PKM2 protein was highly detected in the urine of diabetic patients but was not excreted in the urine of normal subjects. Therefore, PKM2 was selected as the new biomarker for the early diagnosis of DN. Our results reflect current knowledge on the role of PKM2 in DN.

Identification of Novel Biomarker for Early Detection of Diabetic Nephropathy.

Diabetic nephropathy (DN) is one of the most common complications of diabetes mellitus. After development of DN, patients will progress to end-stage renal disease, which is associated with high morbidity and mortality. Here, we developed early-stage diagnostic biomarkers to detect DN as a strategy for DN intervention. For the DN model, Zucker diabetic fatty rats were used for DN phenotyping. The results revealed that DN rats showed significantly increased blood glucose, blood urea nitrogen (BUN), and serum creatinine levels, accompanied by severe kidney injury, fibrosis and microstructural changes. In addition, DN rats showed significantly increased urinary excretion of kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL). Transcriptome analysis revealed that new DN biomarkers, such as complementary component 4b (C4b), complementary factor D (CFD), C-X-C motif chemokine receptor 6 (CXCR6), and leukemia inhibitory factor (LIF) were identified. Furthermore, they were found in the urine of patients with DN. Since these biomarkers were detected in the urine and kidney of DN rats and urine of diabetic patients, the selected markers could be used as early diagnosis biomarkers for chronic diabetic nephropathy.

Optical sensor based on two-dimensional photonic crystals for measuring glucose in urine

The design and simulation of two bio-optical sensors for the noninvasive measurement of glucose levels in urine of diabetic patients are presented. While the sensors are of the same shape and structure, their quality factors (QFs), sensitivities, and sizes are different. The structure developed is based on two-dimensional (2D)-hexagonal-latticed photonic crystals containing silicon rods immersed in urine. By creating line and point defects in the structure, two resonant modes are localized in the photonic bandgap (the photonic bandgap is a frequency range corresponding to mirror action for incident light of frequency within that range). Sensor 1 design was based on localized resonant mode 1, while sensor 2 uses localized resonant mode 2. The mechanism of these sensors relies on the change of the refractive index of urine as a result of the change in glucose concentration, which in turn causes shifts in resonant modes of the sensors. For tuning in the wavelength of 1.55 μm, the scaling method is utilized. The major difference between the proposed sensors and previously reported bio-optical sensors is the determination of glucose level based on the shift in resonant wavelengths. Previous works relied on the change of the amplitude of emerging waves for the same purpose. Consequently, the sensors presented here exhibit a higher QF and operate within the wavelength window of 1.55 μm. QF calculations and sensitivity simulations for sensor 1 yielded approximate values of 3800 and 500 nm/refractive index unit (RIU), respectively. For sensor 2, QF and sensitivity were 5540 and 500 nm / RIU, respectively. The sizes of sensor 1 and sensor 2 are 14.93 × 8.8 μm2 and 15.7 × 9.47 μm2, respectively. The 2D finite difference time domain and plane wave expansion methods were relied upon in the simulations.

Cluster and Factor Analysis of Elements in Serum and Urine of Diabetic Patients with Peripheral Neuropathy and Healthy People

Diabetic peripheral neuropathy (DPN) is a common complication of diabetes mellitus, presented as a major teratogenic cause worldwide. This study discussed alternation and correlation of magnesium (Mg), calcium (Ca), copper (Cu), zinc (Zn), iron (Fe), chromium (Cr), and selenium (Se) among DPN patients and healthy people using multivariate statistical analysis. Fifty patients with DPN were recruited from endocrinology department, First Hospital of Jilin University between January 2010 and October 2011 and also 50 healthy subjects were enrolled at the same time. Inductively coupled plasma mass spectrometry (ICP-MS) was used to assay elements in serum and urine. Cluster analysis was used to clarify alternation of elements’ homogeneity. Factor analysis was performed to evaluate the most informative kinds of elements. Mg, Ca, Zn, and Cr in DPN patients were significantly lower in serum whereas significantly higher in urine. Elements were clustered into 4 or 5 clusters based on internal association using between-groups linkage algorithm. Serum Cr, Se, and Fe were grouped, and Mg related to Ca more closely in both serum and urine in DPN. Factor analysis revealed discrepancies of elements’ contribution. Cr, Se, and Fe appeared to be the most crucial factors contributing to DPN. Mg, Ca, Zn, and Cu were more influential, whereas Cr became less potent to disease. Contributed value of elements could be determined and specified using loadings in scree plot. Future studies and delicate statistical models should be applied.

GRP78 protects CHO cells from ribosylation

d-Ribose (Rib), a reactive glycation compound that exists in organisms, abnormally increases in the urine of diabetic patients and can yield large amounts of advanced glycation end products (AGEs), leading to cell dysfunction. However, whether cellular proteins are sensitive to this type of glycation is unknown. In this study, we found that cellular AGEs accumulate in Chinese hamster ovary (CHO) cells with increased Rib concentration and administration time. Mass spectrum analysis of isolated AGE-modified proteins from cell lysates showed that glucose-regulated protein 78 kD (GRP78) is one of the main ribosylated proteins. Co-immunoprecipitation assays further confirmed the interaction between AGEs and GRP78. Compared with d-glucose (Glc), Rib produced much more AGEs in cells. In kinetic studies, the first order rate constant of LDH released from CHO cells incubated with Rib was nearly 8-fold higher than that of Glc, suggesting that Rib is highly cytotoxic. Immunofluorescent co-localization analysis manifested partial superimposition of AGEs and GRP78, which were distributed throughout the endoplasmic reticulum. Western blotting showed that the expression of GRP78 is up-regulated and then down-regulated in CHO cells during Rib treatment. In the presence of Rib, the suppression of GRP78 expression either with transfected siRNA or with the inhibitor (-)-epigallocatechin gallate (EGCG) dramatically increased AGE levels and decreased cell viability compared with these parameters in the control groups. GRP78 overexpression decreased AGE levels and rescued the cells from Rib-induced cytotoxicity. These data indicate that GRP78 plays a role in preventing Rib-induced CHO cell cytotoxicity.

Urinary beta-trace protein gene expression analysis in type 2 diabetes mellitus patients.

ABSTRACTObjectiveTo evaluate the gene expression of beta-trace protein in urine of diabetic patients, with no reduction in glomerular filtration rate, which was defined as below 60mL/min/1.73m2.MethodsType 2 diabetes mellitus patients were recruited, and a group of non-diabetic individuals served as control. Beta-trace protein gene expression was analyzed by quantitative PCR. Blood samples were collected to establish glucose levels and baseline kidney function. Accuracy was analyzed using ROC curves.ResultsNinety type 2 diabetes mellitus patients and 20 non-diabetic individuals were recruited. The area under the curve was 0.601, sensitivity of 20% and specificity of 89.47%. Among diabetic participants, 18% showed an expression above the cutoff point.ConclusionThese results of accuracy of beta-trace protein gene expression in urine of diabetic patients are promising, although they did not achieve a higher area under the curve level.

GRP78 is a Major Ribosylated Protein in CHO Cells

Over 200 year ago, Chevreul (1786-1889) discovered that the sweetness in the urine of diabetics comes from grape sugar or D-glucose. Glycation with D-glucose leads to the formation of advanced glycation end products (AGEs), which is an important biochemical abnormality that accompanies diabetes mellitus. As one of the most reactive glycation reagents, D-ribose, which can yield large amounts of AGEs in vivo and in vitro, has long been ignored in Diabetes studies. In this laboratory, we have observed abnormally high levels of D-ribose in the urine of diabetic patients, compared with those of age-matched normal control. Our results suggested that diabetes mellitus is not only suffered from dysmetabolism of D-glucose, but also dysmetabolism of D-ribose. Here, we show that treatment with D-ribose in Chinese hamster ovary cells increased expression of cellular AGEs along with the increase of D-ribose concentration and administration time. Mass spectrum analysis of isolated AGE-modified proteins from cell lysates showed glucose-regulated protein 78 kD (GRP78) as a ribosylated protein. Co-immunoprecipitation assay further confirmed interaction between AGEs and GRP78. Immunofluorescence co-localization analysis manifested partial superimposition of AGEs and GRP78. As showed in Western blotting results, changes in GRP78 protein levels in cells exhibited in a time dependent manner in the D-ribose treatment. Inhibition of GRP78 expression with RNAi or function in D-ribose-treated cells both dramatically increased the level of AGEs. The findings from Chinese hamster ovary cells indicated that GRP78 is a major component response to ribosylation playing in protection role for cells in the presence of D-ribose.

Soluble CD36 in plasma and urine: A plausible prognostic marker for diabetic nephropathy

AimsThis study was designed to analyze the level of soluble CD36 (sCD36) in both plasma and urine of type 2 diabetic patients with and without microalbuminuria/macroalbuminuria. MethodsStudy subjects (n=20 each) comprised of those with normal glucose tolerance, type 2 diabetes (T2DM) with normoalbuminiria, T2DM with microalbuminuria and T2DM with macroalbuminuria. The biochemical parameters were analyzed using auto-analyzer, and the level of sCD36 was estimated using an in-house Sandwich ELISA. ResultsThe presence of sCD36 has been identified for the first time in the urine sample. Significant increase in the level of sCD36 was observed in both plasma and urine of diabetic patients with microalbuminuria (P<0.01) and macroalbuminuria (P<0.001). Positive correlation of sCD36 with the kidney markers such as urea, creatinine and eGFR confirmed the association of sCD36 with kidney damage in diabetic patients. Microalbuminuria, which is clinically used as a biomarker for nephropathy showed a strong positive correlation with urine sCD36 (r=0.642; P<0.001) and plasma sCD36 (r=0.498; P<0.001) in Pearson correlation analysis, which was further substantiated in stepwise multiple regression analysis. ConclusionsOur study implies a plausible prognostic/adjuvant biomarker role of soluble CD36 for diabetic nephropathy.

Abstract 657: Increased Urinary Angiotensin Converting Enzyme 2 (ACE2) In Type 2 Diabetic Patients

Diabetes and its associated chronic kidney disease (CKD) is a major health burden and there is an urgent need for new sensitive biomarkers to detect and monitor the progression of CKD. Albuminuria is still the gold standard for the evaluation of kidney function. However, its sensitivity and reliability have recently been questioned. ACE2 is highly expressed in renal tubules and has been shown to be shed in the urine of diabetic patients with CKD. The aim of the study was to investigate whether urinary ACE2 is increased in diabetic patients with CKD before the onset of microalbuminuria. Participants were recruited from Dayton VA Medical Center (Dayton, OH, USA). Baseline urinary albumin creatinine ratio (UACR) and estimated glomerular filtration rate (eGFR) were determined three months before initiation of the study in non-diabetic patients (UACR &lt;30 mg/g, eGFR=97.40±16 ml/min/1.73 m 2 ), and in diabetic patients with normoalbuminuria (UACR &lt;30 mg/g, eGFR=83.08±17 ml/min/1.73 m 2 ), microalbuminuria (UACR = 30-300 mg/g, eGFR=47.13±23 ml/min/1.73 m 2 ), and macroalbuminuria (UACR &gt;300 mg/g, eGFR=39.68±20 ml/min/1.73 m 2 ). Using fluorogenic and mass spectrometry-based enzyme assays, we measured urinary and plasma ACE2 activity in patients. Urinary ACE2 activity was significantly increased in diabetic patients with normoalbuminuria (0.58±0.2 nmol/hr/mg creatinine), microalbuminuria (1.19 ±0.5 nmol/hr/mg creatinine), and macroalbuminuria (2.265±0.4 nmol/hr/mg creatinine) compared with non-diabetic controls (0.06 ± 0.02 nmols/hr/mg creatinine) (p&lt;0.0001). These results were confirmed by detecting the ACE2 product Ang-(1-7) ( m/z 899) in incubations of urine samples with the natural substrate Ang II ( m/z 1046) using mass spectrometry-based enzyme assays. In addition, urinary ACE2 expression was significantly increased in diabetic patients as determined by western blot analysis (p&lt;0.05). Plasma ACE2 activity was not detectable in control and diabetic patients. In conclusion, urinary ACE2 is increased in diabetic patients with CKD which suggests that urinary ACE2 could be used as an early, noninvasive biomarker for diabetic nephropathy before the onset of microalbuminuria.

A novel nanogold–single wall carbon nanotube modified sensor for the electrochemical determination of 8-hydroxyguanine, a diabetes risk biomarker

An electrochemical study of the oxidation of 8-hydroxyguanine (8-OH-Gua) at gold nanoparticles attached to single walled carbon nanotube modified edge plane pyrolytic graphite electrode (AuNP-SWCNT/EPPGE) has been carried out to develop a method for the self diagnosis of diabetes. The level of 8-OH-Gua, an important biomarker of oxidative DNA damage, is higher in urine of diabetic patients than control subjects. A detailed comparison has been made between the square wave voltammetric (SWV) response of SWCNT/EPPGE and AuNP-SWCNT/EPPGE towards the oxidation of 8-OH-Gua in respect of several essential analytical parameters viz. sensitivity, detection limit, peak current and peak potential. The AuNP-SWCNT/EPPGE exhibited a well defined anodic peak at potential of ~221mV for the oxidation of 8-OH-Gua as compared to ~312mV using SWCNT/EPPGE at pH=7.2. Under optimized conditions linear calibration curve for 8-OH-Gua is obtained over a concentration range of 0.01–10.0nM in phosphate buffer solution (PBS) of pH=7.2 with detection limit and sensitivity of 5.0 (±0.1) pM and 4.9 (±0.1) μAnM−1, respectively. The oxidation of 8-OH-Gua occurred in a pH dependent process and the electrode reaction followed adsorption controlled pathway. The electrode exhibited an efficient catalytic response with good reproducibility and stability. The method has been found selective and successfully implemented for the determination of 8-OH-Gua in urine samples of diabetic patients.

D-ribosylation induces cognitive impairment through RAGE-dependent astrocytic inflammation

Non-enzymatic glycation of proteins by reducing saccharides for instance D-glucose is an important post-translational modification regulating protein function. Already two centuries ago, D-glucose (Glc) was identified in the urine of diabetic patients. Recently, abnormally high level of D-ribose (Rib) in the urine of type 2 diabetics has been discovered, which is highly active in protein glycation, resulting in the production of advanced glycation end products (AGEs). Accumulation of AGEs leads to altered cellular function, for example AGE accumulation in the nervous system impairs cognitive ability, yet the mechanisms mediating this process for Rib are unknown. Here we found that treatment with Rib accelerated AGE formation in U251 and U87MG astrocytoma cells and in mouse brain, inducing upregulation of receptor for AGEs (RAGE). Astrocytoma cells with elevated levels of RAGE displayed enhanced activity of the proinflammatory nuclear transcription factor kappaB and increased expression of tumor necrosis factor alpha and glial fibrillary acidic protein. Moreover, injection of Rib induced astrocyte activation in mouse hippocampus and impaired spatial learning and memory abilities. These results indicate that mouse spatial cognitive impairment caused by Rib-derived AGEs is correlated with activation of an astrocyte-mediated, RAGE-dependent inflammatory response. This study may provide insights into the mechanism of Rib-involved cognitive impairments and diabetic encephalopathy.

D-ribose, an overlooked player in type 2 diabetes mellitus?

Type 2 diabetes mellitus is a metabolic disorder that is characterized by high blood glucose due to either insulin resistance or insulin deficiency [1]. A direct correlation between D-glucose and diabetic complications has long been established, and is the focus of most research in this field. In contrast, D-Ribose has been overlooked so far as a potential risk player in the development of diabetes. Recently published reports found abnormally high levels of D-ribose in the urine of diabetic patients [2]. Elsewhere, intravenous administration of D-ribose resulted in a decrease in blood D-glucose, concomitantly with an increase in the level of glycated serum proteins [3]. It is known that administration of high dose of D-ribose rapidly glycates proteins in vivo and in vitro [46], which results in the production of high levels of advanced glycation end products. D-glucose, in contrast, cannot be used to the same effect, even under identical conditions [7]. Therefore, as we hypothesized, dysfunction of the metabolism of D-ribose may play a role involving the development of complications of type 2 diabetes mellitus, and it is feasible that D-ribose could be used as a biomarker for type 2 diabetes mellitus. That is, diabetics suffer from dysfunction of energy metabolism, involving not only Dglucose, but also D-ribose. This viewpoint for D-ribose in diabetes is based on these observations: (i) Diabetics display abnormal increases in the level of uric D-ribose; (ii) Raised levels of D-ribose result in increased ribosylation activity, in the result of which more advanced glycation end products accumulate in cells; and (iii) D-ribose could be used as a new biomarker for type 2 diabetes mellitus, as it is easily measured in the urine of diabetic patients. We suggest that this novel view of type 2 diabetes mellitus deserves further and detailed investigation, both at the level of basic and clinical research, to clarify whether and how D-ribose and its metabolism are related with diabetic complications.

Abstract 158: Role of Prorenin Receptor in the Enhancement of Renin Activity in the Urine of Diabetic Patients

High prorenin levels in the circulation are associated with the progression of diabetic nephropathy and microvascular changes in diabetes. In spite of high plasma prorenin, the urine of diabetes mellitus patients exhibits very low pH and contains high levels of “active” renin, but low levels of prorenin. Prorenin binding to the prorenin receptor (PRR) results in non-proteolytic activation of prorenin and stimulation of intracellular synthesis of transforming growth factor-beta (TGF-β). Furthermore, the PRR cytosolic domain is associated with the vacuolar H+-ATPase, which is involved in acid-base homeostasis by renal intercalated cells. In diabetes, the collecting duct is the major source of prorenin and PRR is upregulated in the kidneys of diabetic rats. We examined the relationships among urine levels of renin (uRen), prorenin (uPro), soluble PRR (sPRR) and TGF-β (uTGF-β) in 93 subjects, including non-diabetes mellitus (Non-DM: n=64); and type-2 diabetes mellitus (DM; n=29) patients. Urine albumin (uAlb), creatinine (uCr) and pH were also assessed. DM patients showed higher uRen levels (DM: 81±21 vs Non-DM: 29±4.5 ng AngI/mL/hr; p&lt;0.001), which were positively associated with uAlb/uCr (p&lt;0.05, r=0.21) and inversely correlated with urine pH (p&lt;0.01, r=-0.38). These values contrasted with uPro levels, which were no different between groups. DM patients showed increased uTGF-β compared to Non-DM (DM: 22±5.3 vs Non-DM: 5.6±1.5 pg/mg uCr; p&lt;0.0001). Urine sPRR levels were positively correlated with urine pH (p&lt;0.0001, r=0.56). No differences were observed in sPRR plasma and urine levels between non-DM and DM patients. However, the plasma levels of sPRR were 12.5 and 18.5-fold higher than in urine of Non-DM and DM, respectively. Treatment with RAS blockers increased sPRR in plasma (DM with RAS blockade: 24,455±2,151 vs DM without RAS blockade: 17,726±1,255 pg/mL, p&lt;0.001) but not in urine. These data suggest that the cell membrane-bound PRR, not the soluble form, might be responsible for the increases in renin activity and TGF-β, as well as in the low pH in the urine of type-2 DM patients. It is likely that prorenin-mediated activation of membrane bound PRR promotes diabetic nephropathy by increasing urinary formation of TGF-β.

The Dissociation of the Soluble Prorenin Receptor in Plasma and Urine of Diabetic Patients

Diabetic patients display low‐to‐normal plasma levels of renin but increased prorenin levels. In contrast, their urine contains high levels of “active” renin but low prorenin. Our hypothesis is that the soluble form of the prorenin receptor (sPRR) contributes to the enhancement renin activity in the urine of diabetic patients. We measured plasma and urinary sPRR levels by enzyme‐linked immunosorbent assays (ELISA; IBL, Japan) in 60 patients, including: Controls (CT; n=20), Diabetics (DM; n=20) and Diabetics with hypertension (DM+HTN; n=20). Plasma sPRR levels were augmented in DM compared to CT and DM+HTN (DM: 9849±828 vs. CT: 7310±715 and DM+HTN: 7121±373 pg/mL; p&lt;0.05). In contrast, in the urine of DM and DM+HTN patients, sPRR levels were decreased (DM: 576.5±130.9 and DM+HTN: 486.1±81.64 vs. CT: 1248.0±345.6 pg/mg; p&lt;0.05) but urinary renin content was increased (DM: 130±14; DM+HTN: 162±31 vs. CT: 10±1.3 ng AngI/mL/hr; p&lt;0.0001). uCre levels were similar among groups. The dissociation between plasma and urine levels of sPRR suggests that lower pH in the urine of diabetic patients may increase the capability of sPRR to enhance renin enzymatic activity. Further studies will be performed to verify it the sPRR is bound or unbound to renin/prorenin in the urine of these patients.

Emergence of Non-albicans Candida in urine of diabetic patients at Gwalior (M.P.), India

The frequent occurrence of Candida infections in patients with diabetes mellitus has been recognized for many years and is thought to be more prevalent among these individuals. The objective of the study was to focus on the isolation of non- albicans Candida in urine of diabetic patients. A total of 80 samples in urine of diabetic patients treated in hospitals, clinics and nursing homes and from pathological laboratories at Gwalior (M.P.), India were recruited for the study. Among the diabetic patients 62.5 % (50/80) had Candida infections. The major Candida species isolated were Candida tropicalis 34% (17) followed by C. albicans 30% (15), C. parapsilosis 16% (08), C. glabrata 14% (07) and C. krusei 6% (03). This study indicates that non- albicans Candida are also emerging in urine of diabetic patients.

Neutrophil Proteinase 3 Induces Diabetes in a Mouse Model of Glucose Tolerance

Type 1 diabetes is considered to be an autoimmune disease in which T cells attack pancreatic islet cells. Impaired glucose tolerance with type 2 diabetes has been classified as an obesity-associated metabolic syndrome. However, recent studies have revealed that type 2 diabetes is an autoinflammatory disease due to an imbalance of inflammatory cytokine production and related molecular components that cause inflammation. Insulin-like growth factor (IGF) and the insulin-like growth factor-binding protein-3 (IGFBP3) system are known to be involved in the development of experimental diabetic nephropathy, and urinary IGFBP3 protease activity has been observed in patients with type 2 diabetes. A serine protease was found to be responsible for the proteolytic activity in diabetic urine; however, the identity of the precise enzyme remains unknown. We investigated neutrophil proteinase 3 (PR3) to see whether it has specific enzymatic activity associated with insulin-like growth factor-1 and IGFBP3. In our study, both molecules were sufficiently degraded, which leads us to believe that PR3 may induce insulin resistance in the mouse model utilized. In addition, we found that PR3 in the urine of diabetic patients similarly affects insulin resistance. Moreover, PR3-immunized mice had an increase in glucose clearance due to inhibition of PR3 activity. As such, PR3 can be considered as an inflammatory enzyme directly linking inflammation to type 2 diabetes through downregulation of insulin-like growth factor-1/IGFBP3.

Antimicrobial activity of antibiotics in urine under different physiological conditions

Methods Urine obtained from healthy volunteers was pooled and sterile filtered. Microdilution tests were performed with Escherichia coli ATCC 25922 in MHB and human urine. The pH of urine was adjusted to values ranging from 5 to 8. For simulating different glucose levels in urine of diabetic patients, urine was adjusted to glucose levels of 100 and 1000 mg/dL. Bacterial growth in different media was investigated by growth curves. Results obtained from MIC testing were confirmed by use of bacterial killing curves. Klebsiella oxytoca ATCC 700324 was used to investigate transferability of finding to other strains. Each experiment was performed 5 times.