Nutrient-dependent regulation of ß-cell proinsulin content.

Insulin is made from proinsulin, but the extent to which fasting/feeding controls the homeostatically regulated proinsulin pool in pancreatic ß-cells remains largely unknown. Here, we first examined ß-cell lines (INS1E and Min6, which proliferate slowly and are routinely fed fresh medium every 2-3 days) and found that the proinsulin pool size responds to each feeding within 1 to 2 h, affected both by the quantity of fresh nutrients and the frequency with which they are provided. We observed no effect of nutrient feeding on the overall rate of proinsulin turnover as quantified from cycloheximide-chase experiments. We show that nutrient feeding is primarily linked to rapid dephosphorylation of translation initiation factor eIF2α, presaging increased proinsulin levels (and thereafter, insulin levels), followed by its rephosphorylation during the ensuing hours that correspond to a fall in proinsulin levels. The decline of proinsulin levels is blunted by the integrated stress response inhibitor, ISRIB, or by inhibition of eIF2α rephosphorylation with a general control nonderepressible 2 (not PERK) kinase inhibitor. In addition, we demonstrate that amino acids contribute importantly to the proinsulin pool; mass spectrometry shows that ß-cells avidly consume extracellular glutamine, serine, and cysteine. Finally, we show that in both rodent and human pancreatic islets, fresh nutrient availability dynamically increases preproinsulin, which can be quantified without pulse-labeling. Thus, the proinsulin available for insulin biosynthesis is rhythmically controlled by fasting/feeding cycles.

Multifunctional self-assembled adsorption microspheres based on waste bamboo shoot shells for multi-pollutant water purification.

In this study, multilayer self-assembled multifunctional bamboo shoot shell biochar microspheres (BSSBM) were prepared, in which bamboo shoot shell biochar was used as the carrier, titanium dioxide as the intermediate medium, and chitosan as the adhesion layer. The adsorption behavior of BSSBM on heavy metals Ag(I) and Pd(II), antibiotics, and dye wastewater was systematically analyzed. BSSBM shows a wide range of adsorption capacity. BSSBM is a promising candidate for the purification of real polluted water, not only for metal ions, but also for Tetracycline (TC) and Methylene Blue (MB). The maximum adsorption amounts of BSSBM on Pd(II), Ag(I), TC and MB were 417.3 mg/g, 222.5 mg/g, 97.2 mg/g and 42.9 mg/g, respectively.The adsorption of BSSBM on Pd(II), MB and TC conformed to the quasi-first kinetic model, and the adsorption on Ag(I) conformed to the quasi-second kinetic model. BSSBM showed remarkable selective adsorption capacity for Ag(I) and Pd(II) in a multi-ion coexistence system. BSSBM not only realized the high value-added utilization of waste, but also had the advantages of low cost, renewable and selective adsorption. BSSBM demonstrated its potential as a new generation of multifunctional adsorbent, contributing to the recovery of rare/precious metals and the treatment of multi-polluted water.

The influence of stereodynamical control on the nonadiabatic effects in the D + HD (v = 1, j = 2) → D2 + H reaction.

Stereodynamics is a field that studies the influence of the alignment or orientation of colliding partners on the results of collisions. At present, the intersection of nonadiabatic effects and stereodynamics remains to be explored. In this study, we theoretically demonstrate significant stereodynamical effects in the D + HD (v = 1, j = 2) → D2 + H reaction within the collision energy range of 0.01-2.99 eV by using the time-dependent wave packet method. It is found that the stereodynamical control not only facilitates the reaction but also allows precise control of the products over a range of different scattering angles. The analysis at the state-to-state level reveals that the nonadiabatic effects are stronger in the parallel configuration than in the perpendicular configuration. By topological approach to separate the two reaction pathways at the conical intersection, the scattering amplitude of the roaming pathway in the parallel configuration is larger than that of the perpendicular configuration, which leads to more dramatic nonadiabatic features in the collision with parallel configuration.

Proteasomal degradation of WT proinsulin in pancreatic beta cells.

Preproinsulin entry into the endoplasmic reticulum yields proinsulin, and its subsequent delivery to the distal secretory pathway leads to processing, storage, and secretion of mature insulin. Multiple groups have reported that treatment of pancreatic beta cell lines, rodent pancreatic islets, or human islets with proteasome inhibitors leads to diminished proinsulin and insulin protein levels, diminished glucose-stimulated insulin secretion, and changes in beta-cell gene expression that ultimately lead to beta-cell death. However, these studies have mostly examined treatment times far beyond that needed to achieve acute proteasomal inhibition. Here, we report that although proteasomal inhibition immediately downregulates new proinsulin biosynthesis, it nevertheless acutely increases beta-cell proinsulin levels in pancreatic beta cell lines, rodent pancreatic islets, and human islets, indicating rescue of a pool of recently synthesized WT INS gene product that would otherwise be routed to proteasomal disposal. Our pharmacological evidence suggests that this disposal most likely reflects ongoing endoplasmic reticulum-associated protein degradation. However, we found that within 60 min after proteasomal inhibition, intracellular proinsulin levels begin to fall in conjunction with increased phosphorylation of eukaryotic initiation factor 2 alpha, which can be inhibited by blocking the general control nonderepressible 2 kinase. Together, these data demonstrate that a meaningful subfraction of newly synthesized INS gene product undergoes rapid proteasomal disposal. We propose that free amino acids derived from proteasomal proteolysis may potentially participate in suppressing general control nonderepressible 2 kinase activity to maintain ongoing proinsulin biosynthesis.

Semidiscrete optical vortex droplets in quasi-phase-matched photonic crystals.

What we believe is a new scheme for producing semidiscrete self-trapped vortices ("swirling photon droplets") in photonic crystals with competing quadratic (χ(2)) and self-defocusing cubic (χ(3)) nonlinearities is proposed. The photonic crystal is designed with a striped structure, in the form of spatially periodic modulation of the χ(2) susceptibility, which is imposed by the quasi-phase-matching technique. Unlike previous realizations of semidiscrete optical modes in composite media, built as combinations of continuous and arrayed discrete waveguides, the semidiscrete vortex "droplets" are produced here in the fully continuous medium. This work reveals that the system supports two types of semidiscrete vortex droplets, viz., onsite- and intersite-centered ones, which feature, respectively, odd and even numbers of stripes, N. Stability areas for the states with different values of N are identified in the system's parameter space. Some stability areas overlap with each other, giving rise to the multistability of states with different N. The coexisting states are mutually degenerate, featuring equal values of the Hamiltonian and propagation constant. An experimental scheme to realize the droplets is outlined, suggesting new possibilities for the long-distance transmission of nontrivial vortex beams in nonlinear media.

Vortex Solitons in Quasi-Phase-Matched Photonic Crystals.

We report solutions for stable compound solitons in a three-dimensional quasi-phase-matched photonic crystal with the quadratic (χ^{(2)}) nonlinearity. The photonic crystal is introduced with a checkerboard structure, which can be realized by means of the available technology. The solitons are built as four-peak vortex modes of two types, rhombuses and squares (intersite- and onsite-centered self-trapped states, respectively). Their stability areas are identified in the system's parametric space (rhombuses occupy an essentially broader stability domain), while all bright vortex solitons are subject to strong azimuthal instability in uniform χ^{(2)} media. Possibilities for experimental realization of the solitons are outlined.

Deficient endoplasmic reticulum translocon-associated protein complex limits the biosynthesis of proinsulin and insulin.

The conserved endoplasmic reticulum (ER) membrane protein TRAPα (translocon-associated protein, also known as signal sequence receptor 1, SSR1) has been reported to play a critical but unclear role in insulin biosynthesis. TRAPα/SSR1 is one component of a four-protein complex including TRAPß/SSR2, TRAPγ/SSR3, and TRAPδ/SSR4. The TRAP complex topologically has a small exposure on the cytosolic side of the ER via its TRAPγ/SSR3 subunit, whereas TRAPß/SSR2 and TRAPδ/SSR4 function along with TRAPα/SSR1 largely on the luminal side of the ER membrane. Here, we have examined pancreatic ß-cells with deficient expression of either TRAPß/SSR2 or TRAPδ/SSR4, which does not perturb mRNA expression levels of other TRAP subunits, or insulin mRNA. However, deficient protein expression of TRAPß/SSR2 and, to a lesser degree, TRAPδ/SSR4, diminishes the protein levels of other TRAP subunits, concomitant with deficient steady-state levels of proinsulin and insulin. Deficient TRAPß/SSR2 or TRAPδ/SSR4 is not associated with any apparent defect of exocytotic mechanism but rather by a decreased abundance of the proinsulin and insulin that accompanies glucose-stimulated secretion. Amino acid pulse labeling directly establishes that much of the steady-state deficiency of intracellular proinsulin can be accounted for by diminished proinsulin biosynthesis, observed in a pulse-labeling as short as 5 minutes. The proinsulin and insulin levels in TRAPß/SSR2 or TRAPδ/SSR4 null mutant ß-cells are notably recovered upon re-expression of the missing TRAP subunit, accompanying a rebound of proinsulin biosynthesis. Remarkably, overexpression of TRAPα/SSR1 can also suppress defects in ß-cells with diminished expression of TRAPß/SSR2, strongly suggesting that TRAPß/SSR2 is needed to support TRAPα/SSR1 function.

Contamination and Biofilm Formation of Foodborne and Opportunistic Pathogens in Yellow-Feathered Chicken Carcass.

This study aims to analyze the contamination and biofilm formation of foodborne and opportunistic pathogens in yellow-feathered chicken carcasses sampled in different seasons and to prove the relationship between biofilm-forming ability and bacterial extracellular polysaccharide (EPS) production. A total of 78 strains were isolated from chicken samples. The strains consisted of 30.8% Escherichia coli, 14.1% Pseudomonas aeruginosa, 12.8% Salmonella enteritidis, 12.8% Klebsiella pnenmoniae, 10.2% Enterobacter cloacae, 8.9% Proteus mirabilis, 5.1% Klebsiella oxytoca, 1.3% Staphylococcus aureus, and 1.3% Citrobacter braakii. Crystal violet staining assay revealed six strains with strong biofilm-forming ability, namely, E. coli S7, K. oxytoca B12, K. pnenmoniae B6, S. enteritidis H4, P. aeruginosa M5, and S. aureus G1, which showed had high abilities of cell motility and EPS production. Confocal laser scanning microscopy and scanning electron microscopy showed that all six strains can form mature biofilm architectures after 5 d of cultivation. This study may serve as a reference to control the contamination of foodborne pathogens in yellow-feathered chicken and enhance the quality and shelf life of these chicken products.

Skin Microbiota in Non-inflammatory and Inflammatory Lesions of Acne Vulgaris: The Underlying Changes within the Pilosebaceous Unit.

Acne vulgaris is a common chronic inflammatory skin disease of the pilosebaceous unit. Clinical manifestations include seborrhea, non-inflammatory lesions, inflammatory lesions, or scar formation. Fourteen eligible participants of either sex, aged 18-28 years old, with mild to moderate acne lesions, were recruited in this observational study. The contents of 10 pilosebaceous units of non-inflammatory (comedones) and inflammatory lesions (papules and pustules) were collected from each participant's face and examined by amplicon metagenomics sequencing and real-time Polymerase Chain Reaction (PCR). Male participants, participants with a higher body mass index (BMI) than normal, and participants younger than 20 years old, were revealed to have a higher proportion of Malassezia in their non-inflammatory lesions than that in inflammatory lesions. There was an increased abundance of Malassezia restricta (M. restricta) and Cutibacterium acnes (C. acnes) in the non-inflammatory group. Correlation analysis indicated that Staphylococcus epidermidis (S. epidermidis) and M. restricta have similar proliferation trends with C. acnes during the transformation from non-inflammatory to inflammatory lesions. M. restricta probably involve in the microecological balance within the pilosebaceous unit.

Misfolded proinsulin impairs processing of precursor of insulin receptor and insulin signaling in ß cells.

Insulin resistance in classic insulin-responsive tissues is a hallmark of type 2 diabetes (T2D). However, the pathologic significance of ß-cell insulin resistance and the underlying mechanisms contributing to defective insulin signaling in ß cells remain largely unknown. Emerging evidence indicates that proinsulin misfolding is not only the molecular basis of mutant INS-gene-induced diabetes of youth (MIDY) but also an important contributor in the development and progression of T2D. However, the molecular basis of ß-cell failure caused by misfolded proinsulin is still incompletely understood. Herein, using Akita mice expressing diabetes-causing mutant proinsulin, we found that misfolded proinsulin abnormally interacted with the precursor of insulin receptor (ProIR) in the endoplasmic reticulum (ER), impaired ProIR maturation to insulin receptor (IR), and decreased insulin signaling in ß cells. Importantly, using db/db insulin-resistant mice, we found that oversynthesis of proinsulin led to an increased proinsulin misfolding, which resulted in impairments of ProIR processing and insulin signaling in ß cells. These results reveal for the first time that misfolded proinsulin can interact with ProIR in the ER, impairing intracellular processing of ProIR and leading to defective insulin signaling that may contribute to ß-cell failure in both MIDY and T2D.-Liu, S., Li, X., Yang, J., Zhu, R., Fan, Z., Xu, X., Feng, W., Cui, J., Sun, J., Liu, M. Misfolded proinsulin impairs processing of precursor of insulin receptor and insulin signaling in ß cells.

Metabolite secretions of Lactobacillus plantarum YYC-3 may inhibit colon cancer cell metastasis by suppressing the VEGF-MMP2/9 signaling pathway.

BACKGROUND: Colorectal cancer (CRC) is a major clinical challenge, and the gut microbiome plays important roles in the occurrence and metastasis of CRC. Lactobacillus and their metabolites are thought to be able to suppress the growth of CRC cells. However, the antimetastatic mechanism of Lactobacillus or their metabolites toward CRC cells is not clear. Therefore, the aim of this study was to assess the inhibitory mechanism of cell-free supernatants (CFSs) of L. rhamnosus GG, L. casei M3, and L. plantarum YYC-3 on metastasis of CRC cells. RESULTS: YYC-3 CFS showed the highest inhibitory effect on CRC cell growth, invasion and migration, and inhibited MMP2, MMP9, and VEGFA gene and protein expression, and protein secretion. Furthermore, it suppressed the activities of MMPs by gelatin zymography. Moreover, the effective compounds in these CFSs were analyzed by Q Exactive Focus liquid chromatography-mass spectrometry. CONCLUSIONS: Our results showed that metabolite secretions of YYC-3 may inhibited cell metastasis by downregulating the VEGF/MMPs signaling pathway. These data suggest that treatment of CRC cells with metabolites from L. plantarum YYC-3 may reduce colon cancer metastasis.

Acne fulminans concomitant with vasculitis-like symptoms mimicking Henoch-Schoenlein purpura.

Acne fulminans is a severe form of acne vulgaris accompanied by systemic symptoms. A 17-year-old Chinese boy presented with an outbreak of necrotic lesions on his face eight days after the onset of palpable purpura, arthralgia, fever, abdominal pain, and proteinuria. He was successfully treated with oral prednisolone and isotretinoin. Vasculitis-like symptoms are rarely reported in acne fulminans; therefore, the physician needs to maintain awareness of this uncommon presentation.

Genome, transcriptome and fermentation analyses of Lactobacillus plantarum LY-78 provide new insights into the mechanism of phenyllactate biosynthesis in lactic acid bacteria.

Phenyllactate (PLA) is found in a variety of fermented foods and is a promising antibacterial agent, drug and plastic synthetic precursor. Previous studies have shown that PLA is a product of Phe catabolism in lactic acid bacteria (LAB), and PLA biosynthesis is mainly related to lactate dehydrogenases (LDHs). Here, the genome, transcriptome and fermentation characteristics of PLA-producing Lactobacillus plantarum LY-78 were studied. The fermentation experiments demonstrated that L. plantarum LY-78 possesses the ability to synthesize PLA de novo. Secondly, the genome and transcriptome analyses revealed candidate pathways, operons and key genes for PLA biosynthesis in the strain. Finally, genome-wide transcriptome analysis revealed significant changes in the expression profile of strain LY-78 in the absence and presence of PPA. Overall, this work demonstrates for the first time that PLA can be a by-product of Phe anabolism in LAB, provides new insights and evidence for elucidating the mechanism of PLA biosynthesis in LAB, and may provide new candidate genes and research strategies for future PLA biosynthesis applications.

Semidiscrete Quantum Droplets and Vortices.

We consider a binary bosonic condensate with weak mean-field (MF) residual repulsion, loaded in an array of nearly one-dimensional traps coupled by transverse hopping. With the MF force balanced by the effectively one-dimensional attraction, induced in each trap by the Lee-Hung-Yang correction (produced by quantum fluctuations around the MF state), stable on-site- and intersite-centered semidiscrete quantum droplets (QDs) emerge in the array, as fundamental ones and self-trapped vortices, with winding numbers, at least, up to five, in both tightly bound and quasicontinuum forms. The application of a relatively strong trapping potential leads to squeezing transitions, which increase the number of sites in fundamental QDs and eventually replace vortex modes by fundamental or dipole ones. The results provide the first realization of stable semidiscrete vortex QDs, including ones with multiple vorticity.

Cord blood leptin DNA methylation levels are associated with macrosomia during normal pregnancy.

BACKGROUND: We previously demonstrated an association between placental leptin (LEP) methylation levels and macrosomia without gestational diabetes mellitus (non-GDM). This study further explored the association between LEP methylation in cord blood and non-GDM macrosomia. METHOD: We carried out a case-control study of 61 newborns with macrosomia (birth weight ≥4000 g) and 69 newborns with normal birth weight (2500-3999 g). Methylation in the LEP promoter region was mapped by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Average cord blood LEP methylation levels were lower in macrosomia newborns than in control newborns (P < 0.001). Eleven CpG sites were associated with macrosomia. Multivariate logistic regression revealed that low LEP methylation levels [adjusted odds ratio (AOR) = 2.84, 95% confidence interval (CI): 1.72-4.17], high pre-pregnancy body mass index (AOR = 7.44, 95% CI: 1.99-27.75), long gestational age (AOR = 3.18, 95% CI: 1.74-5.79), high cord blood LEP concentration (AOR = 2.25, 95% CI: 1.34-3.77), and male newborn gender (AOR = 3.91, 95% CI: 1.31-11.69) significantly increased the risk of macrosomia. CONCLUSIONS: Lower cord blood LEP methylation levels and certain maternal and fetal factors are associated with non-GDM macrosomia.

The In Vitro Protective Role of Bovine Lactoferrin on Intestinal Epithelial Barrier.

The intestinal epithelial barrier plays a key protective role in the gut lumen. Bovine lactoferrin (bLF) has been reported to improve the intestinal epithelial barrier function, but its impact on tight junction (TJ) proteins has been rarely described. Human intestinal epithelial crypt cells (HIECs) were more similar to those in the human small intestine, compared with the well-established Caco-2 cells. Accordingly, both HIECs and Caco-2 cells were investigated in this study to determine the effects of bioactive protein bLF on their growth promotion and intestinal barrier function. The results showed that bLF promoted cell growth and arrested cell-cycle progression at the G2/M-phase. Moreover, bLF decreased paracellular permeability and increased alkaline phosphatase activity and transepithelial electrical resistance, strengthening barrier function. Immunofluorescence, western blot and quantitative real-time polymerase chain reaction revealed that bLF significantly increased the expression of three tight junction proteins-claudin-1, occludin, and ZO-1-at both the mRNA and protein levels, and consequently strengthened the barrier function of the two cell models. bLF in general showed higher activity in Caco-2 cells, however, HIECs also exhibited desired responses to barrier function. Therefore, bLF may be incorporated into functional foods for treatment of inflammatory bowel diseases which are caused by loss of barrier integrity.

Oral Escherichia coli expressing IL-35 meliorates experimental colitis in mice.

BACKGROUND: Ulcerative colitis (UC) is a type of inflammatory bowel disease (IBD) characterized by chronic inflammation of colon. It is commonly believed that the imbalance of immune system and overwhelming production of cytokines are involved in the pathogenesis of UC. Recent studies demonstrated that interleukin-35 (IL-35), a key player in the regulation of inflammation, has been identified as potential therapeutic target to treat UC. However, conventional intravenous administration is costly and inconvenient. The present study was designed to establish a novel IL-35 delivery system and investigate its therapeutic effects on dextran sulfate sodium (DSS)-induced experimental colitis in mice for the first time. METHODS: An engineered Escherichia coli (E. coli/IL-35) expressing IL-35 was constructed. Adult male BALB/c mice randomly got the oral administration of E. coli/IL-35, empty plasmid-transformed E. coli (E. coli0) or PBS for treatment following ingestion of 3% DSS solution for 5 days. Normal mice were used as control group. Colonic and splenic tissues were collected on day 10 post-DSS-induction. Clinical signs, disease activity index (DAI), pathological and immunohistological changes, cytokine profiles and cell populations were evaluated. RESULTS: Intragastric administration of E. coli/IL-35 effectively protected the colitis mice from DSS assimilation including weight loss and colon shortening. Pathological analysis showed significantly lower DAI score and much less intra-colon infiltration of neutrophils and CD3+ cells in the IL-35 treated group. Moreover, E. coli/IL-35-treated mice demonstrated much less CD4+ IL-17A+ Th17 cells and a higher level of CD4+CD25+Foxp3+ Tregs in spleen and mesenteric lymph nodes, as well as increased colon and serum level of IL-10 and IL-35 and decreased levels of IL-6. CONCLUSIONS: Our study showed that E. coli/IL-35 as a novel oral IL-35 delivery system alleviated inflammatory damage of colonic tissue in the colitic mice. Genetic therapeutic strategies using engineered E. coli encoding immunoregulatory cytokines may provide a potential approach for the treatment of IBD.

NSCLC harboring EGFR exon-20 insertions after the regulatory C-helix of kinase domain responds poorly to known EGFR inhibitors.

Anecdote clinical observations hint that non-small cell lung cancer (NSCLC) with exon-20 insertions might respond poorly to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), contrasting to those with classic mutations. Lack of patient-derived experimental models has been a major hurdle for the discovery of new treatment for the diseases. We established two NSCLC-PDXs harboring two different exon-20 insertions, LU0387-adenocarcinoma (ADC) with a nine-base insertion at 2319 (H773-V774insNPH) and LU3075-squamous cell carcinoma (SCC) with a nine-base insertion at 2316 (P772-H773insDNP). Both insertions immediately follow the regulatory C-helix of the kinase domain. Contrary to the generally good responses to EGFR inhibitors observed in PDXs with classic mutations, both exon-20 insertions are largely resistant to cetuximab and TKIs in vivo, suggesting fundamental difference from the classic EGFR mutations, consistent with the poor response rate to TKI seen in anecdotal clinic reports. It is worth noting that although responses are generally poor, they differ between the two exon-20 mutants depending on the type of TKI. In vitro drug sensitivity assays using established primary cell lines from our two PDXs largely confirmed the in vivo data. Our data from patient-derived experimental models confirmed that exon-20 insertions in domain immediately following the C-helix confer poor response to all known EGFR inhibitors, and suggested that these models can be utilized to facilitate the discovery of new therapies targeting NSCLC harboring exon-20 insertions.

Human endometrial regenerative cells attenuate renal ischemia reperfusion injury in mice.

BACKGROUND: Endometrial regenerative cells (ERCs) is an attractive novel type of adult mesenchymal stem cells that can be non-invasively obtained from menstrual blood and are easily replicated at a large scale without tumorigenesis. We have previously reported that ERCs exhibit unique immunoregulatory properties in experimental studies in vitro and in vivo. In this study, the protective effects of ERCs on renal ischemia-reperfusion injury (IRI) were examined. METHODS: Renal IRI in C57BL/6 mice was induced by clipping bilateral renal pedicles for 30 min, followed by reperfusion for 48 h. ERCs were isolated from healthy female menstrual blood, and were injected (1 million/mouse, i.v.) into mice 2 h prior to IRI induction. Renal function, pathological and immunohistological changes, cell populations and cytokine profiles were evaluated after 48 h of renal reperfusion. RESULTS: Here, we showed that as compared to untreated controls, administration of ERCs effectively prevented renal damage after IRI, indicated by better renal function and less pathological changes, which were associated with increased serum levels of IL-4, but decreased levels of TNF-α, IFN-γ and IL-6. Also, ERC-treated mice displayed significantly less splenic and renal CD4(+) and CD8(+) T cell populations, while the percentage of splenic CD4(+)CD25(+) regulatory T cells and infiltrating M2 macrophages in the kidneys were significantly increased in ERC-treated mice. CONCLUSIONS: This study demonstrates that the novel anti-inflammatory and immunoregulatory effects of ERCs are associated with attenuation of renal IRI, suggesting that the unique features of ERCs may make them a promising candidate for cell therapies in the treatment of ischemic acute kidney injury in patients.