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Objective: Pharmacological inhibition of angiotensin II is currently the only specific therapeutic approach known to slow the course of progressive chronic renal fibrosis independent of the underlying disorder. Since renal fibrosis and atherosclerosis share a number of cellular and molecular features and given the documented benefits of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) in vascular disease, we investigated the effect of rosuvastatin, a very effective statin, in a rat model of chronic progressive glomerulosclerosis (cGS). Methods: cGS was induced by murine anti-thy1 antibody injection into uni-nephrectomized male Wistar rats (n = 8 per group). In cGS, the initial brief anti-thy1 antibody-mediated glomerular injury slowly progresses towards glomerulosclerosis, tubulointerstitial fibrosis and renal failure. Treatment with rosuvastatin (10mg/kg body weight, with food) was started 7 days after cGS induction and continued until week 20. Results: Compared to the untreated cGS rats, administration of rosuvastatin significantly limited the renal disease progression, as shown by reduced proteinuria (314 ± 56 vs 589 ± 51 mg/day), blood pressure (132 ± 5 vs 149 ± 3mm Hg), tubulointerstitial matrix expansion (matrix score 1.7 ± 0.5 vs 3.2 ± 0.3), cortical protein expression of transforming growth factor-beta1 (228 ± 33 vs 483 ± 53 pg/mL) and fibronectin (3,357 ± 647 vs 7,568 ± 1,326 ng/mL), as well as improved renal function (plasma creatinine 0.78 ± 0.2 vs 1.28 ± 0.2 mg/dL, plasma urea 80 ± 17 vs 137 ± 15 mg/dL) (all p < 0.05 cGS + rosuvastatin vs cGS). Glomerular matrix protein expression and accumulation was moderately lower in the statin group. Twelve-hour fasting total plasma cholesterol levels were significantly reduced by rosuvastatin (141 ± 11 vs 198 ± 14 mg/dL, p < 0.001). Conclusions: Rosuvastatin limits the progression of anti-thy 1-induced glomerulonephritis towards chronic tubulointerstitial fibrosis and renal insufficiency. The results suggest that rosuvastatin may directly interfere with pathways mediating progression of chronic kidney disease. Further studies are needed to determine whether rosuvastatin co-therapy acts additively to angiotensin II inhibition. [ABSTRACT FROM AUTHOR]

Abstract Background Epidermal growth factor receptor (EGFR) signaling plays an important role in non-small cell lung cancer (NSCLC) and therapeutics targeted against EGFR have been effective in treating a subset of patients bearing somatic EFGR mutations. However, the cancer eventually progresses during treatment with EGFR inhibitors, even in the patients who respond to these drugs initially. Recent studies have identified that the acquisition of resistance in approximately 50% of cases is due to generation of a secondary mutation (T790M) in the EGFR kinase domain. In about 20% of the cases, resistance is associated with the amplification of MET kinase. In the remaining 30-40% of the cases, the mechanism underpinning the therapeutic resistance is unknown. Methods An erlotinib resistant subline (H1650-ER1) was generated upon continuous exposure of NSCLC cell line NCI-H1650 to erlotinib. Cancer stem cell like traits including expression of stem cell markers, enhanced ability to self-renew and differentiate, and increased tumorigenicity in vitro were assessed in erlotinib resistant H1650-ER1 cells. Results The erlotinib resistant subline contained a population of cells with properties similar to cancer stem cells. These cells were found to be less sensitive towards erlotinib treatment as measured by cell proliferation and generation of tumor spheres in the presence of erlotinib. Conclusions Our findings suggest that in cases of NSCLC accompanied by mutant EGFR, treatment targeting inhibition of EGFR kinase activity in differentiated cancer cells may generate a population of cancer cells with stem cell properties.

Abstract Background The yeast cell cycle is largely controlled by the cyclin-dependent kinase (CDK) Cdc28. Recent evidence suggests that both CDK complex stability as well as function during mitosis is determined by precise regulation of Swe1, a CDK inhibitory kinase and cyclin binding partner. A model of mitotic progression has been provided by study of filamentous yeast. When facing nutrient-limited conditions, Ras2-mediated PKA and MAPK signaling cascades induce a switch from round to filamentous morphology resulting in delayed mitotic progression. Results To delineate how the dimorphic switch contributes to cell cycle regulation, temperature sensitive cdc28 mutants exhibiting constitutive filamentation were subjected to epistasis analyses with RAS2 signaling effectors. It was found that Swe1-mediated inhibitory tyrosine phosphorylation of Cdc28 during filamentous growth is in part mediated by Ras2 activation of PKA, but not Kss1-MAPK, signaling. This pathway is further influenced by Cks1, a conserved CDK-binding partner of elusive function with multiple proposed roles in CDK activation, transcriptional regulation and ubiquitin-mediated proteasome degradation. Conclusion The dynamic balance between Cks1- and Swe1-dependent regulation of Cdc28 and, thereby, the timing of mitosis during yeast dimorphism is regulated in part by Ras2/cAMP-mediated PKA signaling, a key pathway controlling filamentous growth.

Objective: Activation of soluble guanylate cyclase (sGC) and generation of cGMP is the main signal transducing event of the L-arginine-nitricoxide (NO) pathway. The present study analyzes the expression and activity of the NO-cGMP signaling cascade in and the effect of the specific sGC stimulator Bay 41-2272 on the early injury and subsequent repair phase of acute anti-thy1 glomerulonephritis. Methods: Anti-thy1 glomerulonephritis (aGN) was induced by OX-7 antibody injection in male Wistar rats. In protocol 1 (injury), Bay 41-2272 was given starting 6 days before antibody injection. One day after disease induction, parameters of mesangial cell injury (glomerular cell number and inducible NO synthesis) were analyzed. In protocol 2 (repair), Bay 41-2272 treatment was started one day after antibody injection. On day 7, parameters of glomerular repair (glomerular matrix score, protein expression of transforming growth factor (TGF)-beta1, fibronectin and plasminogen-activatorinhibitor (PAI)-1) were determined. In both protocols, tail bleeding time, systolic blood pressure, plasma cGMP levels, glomerular mRNA expression of alpha1 and beta1 sGC and NO-stimulated glomerular cGMP production were analyzed. Results: Bay 41-2272 prolonged bleeding time (+76%), reduced systolic blood pressure (-22mm Hg) and increased plasma cGMP levels (+60%) in both protocols (all p < 0.05 vs. aGN). In the injury experiment, disease induction increased inducible NO synthesis (+250%) and reduced glomerular cell number (-12.3 cell nuclei/glomerular cross section), while expression (alpha1 sGC -94%, beta1 sGC -90%) and activity (-80%) of the soluble guanylate cyclase was almost completely diminished (all p < 0.01 vs. normal control). Bay 41-2272 did not affect parameters of mesangial cell injury and glomerular sGC expression and activity. In the repair protocol, expression (alpha1 sGC +231%, beta1 sGC +463%) and activity (+20,875%) of the soluble guanylate cyclase was markedly increased by disease (all p < 0.01 vs. normal control). Bay 41-2272 further enhanced glomerular sGC expression (alpha1 sGC +1,117%, beta1 sGC +1,658%) and activity (+101120%) (all p < 0.001 vs. normal control and p < 0.05 vs. aGN). This went along with significant reductions in proteinuria (-37%), glomerular matrix accumulation (-18%) and expression of TGF-beta1 (-29%), fibronectin (-39%) and PAI-1 (-22%) as compared to the untreated anti-thy1 animals (all p < 0.05 vs. aGN). Conclusions: Glomerular NO signaling via cGMP is markedly impaired during injury of anti-thy1 glomerulonephritis, while it is highly upregulated during subsequent repair. Further pharmacological sGC stimulation limits glomerular TGF-beta overexpression and matrix expansion. The results suggest that the soluble guanylate cyclase enzyme represents an important anti-fibrotic pathway in glomerular disease. [ABSTRACT FROM AUTHOR]

Current online hemodiafiltration devices can be used to determine the absolute blood volume in clinical practice using the dialysate bolus method. Most of publications on this method have focused on preventing intradialytic complications. The influence of absolute blood volume on long-term prognosis has not been reported yet. A total of 79 participants in a previous study about absolute blood volume were followed for 5 years. Patients with a specific blood volume above ( n  = 45) and below 75 ml/kg ( n  = 34) respectively were compared with regard to survival using Kaplan-Meier analysis. Patients with a specific blood volume below 75 ml/kg had a significantly higher overall 5-year survival rate than patients above 75 ml/kg (70% vs 39%, p  = 0.0233). In patients without cardiac dysfunction, there were no significant differences in 5-year survival between a specific blood volume below or above 75 ml/kg (66% vs 51%). A specific blood volume above 75 ml/kg was associated with an increased mortality in patients with mildly impaired left-ventricular systolic ejection fraction of 40%-59%, whereas in patients with normal blood volume this cardiac impairment did not impact mortality (22% vs 90% 5-year survival, p  = 0.0036). This demonstrates the significance of optimum volume control for long-term survival particularly in cases of reduced cardiac function., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Next-generation sequencing (NGS) technologies have expanded the spectrum of forensic DNA analysis by facilitating efficient and precise genotyping of a large number of genetic markers. Yet, challenges persist regarding complex sample processing and assurance of equal molar concentrations across pooled samples. Since optimal cluster density is crucial for sequencing performance, the determination of both quantity and quality is indispensable for library preparation. In this study, we investigated the application of the Agilent 2100 Bioanalyzer for library quality control, as studies for forensic approaches, particularly for highly degraded postmortem samples, are rare. Our analysis encompassed assessing total DNA concentrations, fluorescence unit (FU) values, and adapter dimer concentrations in purified DNA libraries derived from buccal swabs and tissue samples of decomposed corpses. The sensitivity study tested a serial dilution derived from buccal swabs and revealed a decrease in FU values and an increase in adapter dimers with declining DNA input concentrations. Deviations in total DNA concentrations and average peak heights between the Agilent 2100 Bioanalyzer runs indicated a lack of repeatability in data and presented challenges in accurate quantification, which was also observed in previous studies. Yet, the analysis of degraded samples from decomposed human remains has shown the ability to detect adapter dimer concentrations, which can be crucial for the quality of subsequent NGS library preparation and sequencing success. Therefore, the Agilent 2100 Bioanalyzer proves to be a valuable tool for NGS quality control., (© 2024 The Author(s). Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Background: The rate and the duration of ultrafiltration (UF) are considered the most important factors to affect vascular refilling. The aim of the study was to investigate whether a UF profile could improve the vascular refilling., Methods: Dialysis was delivered by a machine providing feedback control of ultrafiltration rates. Absolute blood volume (BV) was measured by dialysate bolus method. Vascular refilling volume ( V ref ) was calculated as UF volume - Δ absolute BV., Results: In 40 patients, refilling fraction ( V ref /UF volume) was 30.5% in the first hour. Thereafter, refilling fraction steeply increased and reached maximum values in the third and fourth hour at about 95%. The cumulative refilling fraction was 68.5 ± 9.4% at the end. In 14 patients, refilling data from the feedback-controlled UF profile were compared to dialysis sessions with constant UF rates. In 12 of 14 patients, refilling fraction was significantly ( p  = 0.013) higher in sessions with UF profile (71.6% vs 64.4%).There was a significant negative correlation ( r  = -0.606; p  = 0.002) between the blood volume to extracellular volume ratio and the refilling fraction. The sum of this ratio and the refilling fraction was 1.01 ± 0.06., Conclusions: Despite significant differences, a feedback-controlled UF profile has no advantage over the previous refilling studies with regard to the refilling fraction because vascular refilling seems to depend mainly on the ratio of BV to ECV. This would explain the different results in studies using BV guided UF feedback programs., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Absolute blood volume can be calculated from the increase in relative blood volume after an infusion of a well-defined volume bolus of ultrapure dialysate into the extracorporeal circulation. Several working groups have applied this method in research and clinical practice. A critical analysis of differing blood volume data between working groups revealed methodologic problems of the measurement procedure and some important technical aspects. This paper presents a statement to standardize the method., Competing Interests: Disclosure: The authors have no conflicts of interest to report., (Copyright © ASAIO 2024.)

Iodine deficiency-induced goiter continues to be a global public health concern, with varying manifestations based on geography, patient's age, and sex. To gain insights into clinical occurrences, a retrospective study analyzed medical records from patients with iodine deficiency-induced goiter or thyroid cancer who underwent surgery at the Community Hospital in Riehen, Switzerland, between 1929 and 1989. Despite today's adequate iodine supplementation, a significant risk for iodine-independent goiter remains in Switzerland, suggesting that genetic factors, among others, might be involved. Thus, a pilot study exploring the feasibility of genetic analysis of blood spots from these medical records was conducted to investigate and enhance the understanding of goiter development, potentially identify genetic variations, and explore the influence of dietary habits and other environmental stimuli on the disease.Blood prints from goiter patients' enlarged organs were collected per decade from medical records. These prints had been made by pressing, drawing, or tracing (i.e., pressed and drawn) the removed organs onto paper sheets. DNA analysis revealed that its yields varied more between the prints than between years. A considerable proportion of the samples exhibited substantial DNA degradation unrelated to sample collection time and DNA mixtures of different contributors. Thus, each goiter imprint must be individually evaluated and cannot be used to predict the success rate of genetic analysis in general. Collecting a large sample or the entire blood ablation for genetic analysis is recommended to mitigate potential insufficient DNA quantities. Researchers should also consider degradation and external biological compounds' impact on the genetic analysis of interest, with the dominant contributor anticipated to originate from the patient's blood., (© 2024. The Author(s).)

Background: The objective of this study was to investigate the utility of neutrophil gelatinase-associated lipocalin (NGAL) and calprotectin (CPT) to predict long-term graft survival in stable kidney transplant recipients (KTR)., Methods: A total of 709 stable outpatient KTR were enrolled >2 months post-transplant. The utility of plasma and urinary NGAL (pNGAL, uNGAL) and plasma and urinary CPT at enrollment to predict death-censored graft loss was evaluated during a 58-month follow-up., Results: Among biomarkers, pNGAL showed the best predictive ability for graft loss and was the only biomarker with an area under the curve (AUC) > 0.7 for graft loss within 5 years. Patients with graft loss within 5 years (n = 49) had a median pNGAL of 304 [interquartile range (IQR) 235-358] versus 182 (IQR 128-246) ng/mL with surviving grafts (P < .001). Time-dependent receiver operating characteristic analyses at 58 months indicated an AUC for pNGAL of 0.795, serum creatinine-based Chronic Kidney Disease Epidemiology Collaboration estimated glomerular filtration rate (eGFR) had an AUC of 0.866. pNGAL added to a model based on conventional risk factors for graft loss with death as competing risk (age, transplant age, presence of donor-specific antibodies, presence of proteinuria, history of delayed graft function) had a strong independent association with graft loss {subdistribution hazard ratio (sHR) for binary log-transformed pNGAL [log2(pNGAL)] 3.4, 95% confidence interval (CI) 2.24-5.15, P < .0001}. This association was substantially attenuated when eGFR was added to the model [sHR for log2(pNGAL) 1.63, 95% CI 0.92-2.88, P = .095]. Category-free net reclassification improvement of a risk model including log2(pNGAL) in addition to conventional risk factors and eGFR was 54.3% (95% CI 9.2%-99.3%) but C-statistic did not improve significantly., Conclusions: pNGAL was an independent predictor of renal allograft loss in stable KTR from one transplant center but did not show consistent added value when compared with baseline predictors including the conventional marker eGFR. Future studies in larger cohorts are warranted., (© The Author(s) 2023. Published by Oxford University Press on behalf of the ERA.)

The ratio of blood volume to extracellular volume is approximately one to three under physiological conditions and also in stable chronic hemodialysis patients. Recently, it was found that this ratio remains unchanged during hemodialysis despite ultrafiltration. This would signify that the higher the ratio, the lower the refilling and vice versa. To test this hypothesis, treatment data of a previous study were re-analyzed. In 79 stable chronic hemodialysis patients, the refilling fraction was 0.749 ± 0.094. There was a significant negative correlation ( r  = -0.412; p  < 0.001) between the blood volume to extracellular volume ratio and the refilling fraction. The blood volume to extracellular volume relationship seems to be a significant determinant of vascular refilling: the higher the ratio, the lower the refilling, and vice versa., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Touch DNA has become increasingly important evidence in todays' forensic casework. However, due to its invisible nature and typically minute amounts of DNA, the collection of biological material from touched objects remains a particular challenge that underscores the importance of the best collection methods for maximum recovery efficiency. So far, swabs moistened with water are often utilized in forensic crime scene investigations for touch DNA sampling, even though an aqueous solution provokes osmosis, endangering the cell's integrity. The aim of the research presented here was to systematically determine whether DNA recovery from touched glass items can be significantly increased by varying swabbing solutions and volumes compared with water-moistened swabs and dry swabbing. A second objective was to investigate the possible effects of storage of swab solutions prior to genetic analysis on DNA yield and profile quality when stored for 3 and 12 months, as is often the case with crime scene samples. Overall, the results indicate that adapting volumes of the sampling solutions had no significant effect on DNA yield, while the detergent-based solutions performed better than water and dry removal, with the SDS reagent yielding statistically significant results. Further, stored samples showed an increase in degradation indices for all solutions tested, but no deterioration in DNA content and profile quality, allowing for unrestricted processing of touch DNA samples stored for at least 12 months. One further finding was a strong intraindividual change in DNA amounts observed over the 23 deposition days which may be related to the donor's menstrual cycle., (© 2023 The Authors. Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Kidney stones (KSs) are very common, excruciating, and associated with tremendous healthcare cost, chronic kidney disease (CKD), and kidney failure (KF). Most KSs are composed of calcium oxalate and small increases in urinary oxalate concentration significantly enhance the stone risk. Oxalate also potentially contributes to CKD progression, kidney disease-associated cardiovascular diseases, and poor renal allograft survival. This emphasizes the urgent need for plasma and urinary oxalate lowering therapies, which can be achieved by enhancing enteric oxalate secretion. We previously identified Oxalobacter formigenes (O. formigenes) -derived factors secreted in its culture-conditioned medium (CM), which stimulate oxalate transport by human intestinal Caco2-BBE (C2) cells and reduce urinary oxalate excretion in hyperoxaluric mice by enhancing colonic oxalate secretion. Given their remarkable therapeutic potential, we now identified Sel1-like proteins as the major O. formigenes -derived secreted factors using mass spectrometry and functional assays. Crystal structures for six proteins were determined to confirm structures and better understand functions. OxBSel1-14-derived small peptides P8 and P9 were identified as the major factors, with P8 + 9 closely recapitulating the CM's effects, acting through the oxalate transporters SLC26A2 and SLC26A6 and PKA activation. Besides C2 cells, P8 + 9 also stimulate oxalate transport by human ileal and colonic organoids, confirming that they work in human tissues. In conclusion, P8 and P9 peptides are identified as the major O. formigenes -derived secreted factors and they have significant therapeutic potential for hyperoxalemia, hyperoxaluria, and related disorders, impacting the outcomes of patients suffering from KSs, enteric hyperoxaluria, primary hyperoxaluria, CKD, KF, and renal transplant recipients. NEW & NOTEWORTHY We previously identified Oxalobacter formigenes -derived secreted factors stimulating oxalate transport by human intestinal epithelial cells in vitro and reducing urinary oxalate excretion in hyperoxaluric mice by enhancing colonic oxalate secretion. We now identified Sel1-like proteins and small peptides as the major secreted factors and they have significant therapeutic potential for hyperoxalemia and hyperoxaluria, impacting the outcomes of patients suffering from kidney stones, primary and secondary hyperoxaluria, chronic kidney disease, kidney failure, and renal transplant recipients.

Touch DNA recovery techniques can have limitations, as their effectiveness depends on the substrate on which the DNA of a person of interest can be found. In this study, an in-house dry-vacuuming device, the DNA-Buster, was compared to traditional methods for its DNA recovery performance from items typically examined in forensic casework. The aim was to evaluate whether this dry-vacuuming approach can recover DNA efficiently, potentially complementing the well-established recovery strategies. For this, the performances of swabbing, taping, wet- (M-Vac®) and dry-vacuuming (DNA-Buster) were investigated quantitatively and qualitatively for touch DNA deposited on carpet, cotton sweater, stone, tile and wood. For the sweater, both vacuuming methods outperformed the other collection tools quantitatively. While the highest DNA amounts for the carpet were yielded by swabbing and taping, dry-vacuuming was equally good in reaching full DNA profiles, whereas less complete profiles were observed for the M-Vac®. For stone and tile, swabbing was optimal, whereas dry-vacuuming clearly underperformed for these substrates. Taping was the best recovery method for wood. Despite applying single donor DNA after thoroughly cleaning the items, undesired DNA mixtures were detected for all recovery techniques and all substrates. The overall research findings show first that the novel dry-vacuuming method is suited for DNA recovery from textiles. Secondly, they indicate that more attention should be paid to the substrate-collection dependency to ensure best practices in recovering genetic material in a precise, confident and targeted manner from the variety of forensic casework material., Competing Interests: Conflict of interests The authors declare they have no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)