Indiana University-Purdue University Indianapolis (IUPUI) / Nephrolithiasis will affect one-in-eleven people, and more than half of those individuals will have stone recurrence within a decade of their first episode. Despite decades of biomedical research on nephrolithiasis and extraordinary advances in molecular and cell biology, the precise mechanisms of kidney stone formation are not fully understood. Currently, there are limited treatments or preventative measures for nephrolithiasis. Therefore, it is crucial to scrutinize kidney stones from a molecular and cell biology perspective to better understand its pathogenesis and pathophysiology; and to, hereafter, contribute to effective therapeutic targets and preventative strategies. Kidney stones are composed of an admixture of crystal aggregated material and an organic matrix. 80% of all kidney stones are composed of calcium oxalate (CaOx) and half of all CaOx patients grow their stones on to Randall’s plaques (RP). RP are interstitial calcium phosphate mineral deposits in the renal papilla. Thus, we developed and optimized methodologies to directly interrogate CaOx stones. CaOx stones were demineralized, sectioned, and imaged by microscopy, utilizing micro CT for precise orientation. Laser microdissection (LMD) of specific regions of stone matrix analyzed by proteomics revealed various proteins involved in inflammation and the immune response. Analyses on jackstone calculi, having arm protrusions that extend out from the body of the stone, revealed that they are a rare subtype of CaOx stone formation. Micro CT analyses on 98 jackstones showed a radiolucent, organic-rich core in the arm protrusions. Fluorescence imaging on RP stones showed consistent differences in autofluorescence patterns between RP and CaOx overgrowth regions. Moreover, cell nuclei were discovered with preserved morphology in RP regions, along with variable expressions of vimentin and CD45. In comparing spatial transcriptomic expression of reference and CaOx kidney papillae, CaOx patients differentially expressed genes associated with pathways of immune cell activation, reactive oxygen damage and injury, extracellular remodeling, and ossification. Our findings provide novel methodologies to better understand the role of molecules and cells in CaOx stone matrix. Several of the proteins and cells identified in these studies may serve as potential biomarkers, and future therapeutic targets in preventing kidney stone disease.

The aim of this study sets out to better understand nephrology nurses’ lived experiences of dying and deaths of patients with ESKD following withdrawal of dialysis. A qualitative research design using an interpretative phenomenological approach was used to explore the experiences of a purposive heterogeneous sample of eight nephrology nurses who were working in private dialysis units. Information was gathered by phenomenological conversations and feed-back sessions. Colaizzi’s phenomenological method was employed to formulate four main themes: 1. Emotional trauma 2. Detachment 3. Loss of altruistic values in nursing 4. being-with-death For the participants in this study, emotional trauma was the most significant. The participants experienced a sense of powerlessness which caused emotions of hopelessness and anger and subsequently a sense of premature mourning and detachment. This state of hopelessness proved to be an obstacle in patient care, resulting in the altruistic values of nursing to be no longer applied. The participants’ ontological confrontation of being-with-death was evident, as they came to terms with the reality of their own death. Recommendations are offered to address the educational aspects of death and dying for nephrology nurses. This study endorses the need for further research into patients with ESKD ‘end-of-life’ which can influence how healthcare professionals should treat these patients during this phase.

Nephrotic syndrome (NS) is the second leading cause of chronic kidney disease (CKD) presenting under the age of 30. NS presents in children with edema and severe proteinuria, caused by the effacement of podocyte foot processes within the glomerular filtration barrier. Patients with steroid-resistant NS (SRNS) frequently develop end-stage renal disease (ESRD). Additionally, renal biopsies from these patients often reveal focal segmental glomerulosclerosis (FSGS). Pathogenic mutations in known monogenic disease genes have been found in 11-45% children with FSGS/SRNS. Notably, most Mendelian etiologies exhibit recessive inheritance, while dominant vertical inheritance with incomplete penetrance is observed in the remainder. The role of de novo variants (DNVs) in NS necessitates further investigation. Tripartite motif containing 8, TRIM8, is an E3 ubiquitin ligase. De novo TRIM8 variants were previously implicated in a syndromic disease consisting of neurodevelopmental delay, epilepsy, cerebral atrophy, and nephrotic syndrome. In this study, we recapitulate the patient-specific mutations in inducible overexpression cell lines and in CRISPR/Cas9-generated mouse models. N-terminal MYC or GFP-tagged TRIM8 inducible cell lines were generated and characterized using the pInducer21 system. Western blot and immunofluorescence data show that MYC- and GFP-TRIM8 were induced by doxycycline in immortalized podocyte cell lines. Candidate interactors for TRIM8 from the literature and stratified using kidney single cell mRNA sequencing expression were cloned into mammalian expression vectors. Finally, a Trim8 knockout allele (c. 56_162del; p.H20Qfs*124 and c.367_463+304delins46) was generated and bred to yield an allelic series of wildtype, heterozygous and homozygous animals. These mice exhibited normal survival and did not demonstrate proteinuria through three to four months of life. Overall, further studies are ongoing with regards to the continued monitoring of proteinuria and kidney dysfunction, as well as the potential interactor cloning and cell line characterization. / 2025-02-06T00:00:00Z

Nursing care in nephrology brings certain specifics both for the patient and for the nursing staff concerned. For each member of a nephrology multidisciplinary team that participates in identification and meeting bio-psycho-social and spiritual needs of dialysed patients, the quality nursing care provision and the patients´ safety assuring should be the rule. To assure quality care provision in a health care facility, it is necessary to define the professional level of quality, i.e. to introduce processual nursing standards at the national level for all dialysis centers. The objective of this thesis was to make a survey of historical milestones in the development of nephrology nursing in the Czech Republic (in the former Czechoslovakia), from its beginning to present days. In individual parts of the theoretical section various contexts of nursing issues (historical, legal, social, relational, educational) and their interaction are described. The thesis also is focused on detection the importance of aspects influencing the quality of provided nursing care in nephrology and realization of analysis the use and compliance with the existing local nursing standards in individual dialysis centers in the Czech Republic. Another objectives of this thesis were detection of satisfaction rate with existing national standards for nephrology nursing care and to make up a proposal of a national processual standard of nursing care provided to adult dialysed patients. To obtain data the mixed methods research was used. The practical part offers, through conclusions of the quantitative research, explanations of many facts by confirmation or refutation of hypotheses arisen from the qualitative research. The nursing research was conducted through a questionnaire survey in dialysis centers in the Czech Republic. The respondents were head nurses - nursing care managers working in these units. Interviews with representatives of the biggest professional organization of nurses {--} the Czech Nurses Association were also carried out as another research procedure. The survey outcomes have clearly demonstrated the need to introduce national processual standards of nursing care in nephrology practice. Description of the present state is the first step for its improvement. The way out from the current situation is the development of general national processual standards for individual areas of care for nephrology patients and subsequent introduction of them into clinical practice in all dialysis centers in the Czech Republic. One of the components of the thesis is a proposal of the national processual standard of nursing care for a dialysed patient with acute or permanent central venous catheter (CVC). This standard could serve as a model for the development of other national standards of nursing care in nephrology.

Intradialytic Hypotension (IDH) is the commonest complication of maintenance haemodialysis and is associated with increased morbidity and mortality. However, there is no standardised definition of IDH, making comparisons between studies difficult. This observational study with a total of 80 patients and over 600 dialysis sessions showed a poor correlation between symptoms and hypotension. Importantly, patients experienced low blood pressure without symptoms, so continuous intradialytic blood pressure monitoring is required to identify this asymptomatic group. In light of these findings, a revised definition of IDH is suggested. This study also aimed to identify predictors of IDH that could be detected in sufficient time to allow a mitigating intervention. A novel non-invasive alternative for continuous blood pressure monitoring is introduced which uses intra-fistula pressure data from the sensors sited in the extracorporeal circuit of the dialysis machine. Results show that in the majority of patients, changes in intra-fistula pressure correlate with blood pressure measurements obtained by a standard oscillometric device. To investigate whether IDH can be predicted, a photoplethysmogram (PPG) waveform was obtained from a pulse oximeter attached to the finger or ear. Continuous PPG monitoring of patients with IDH during dialysis demonstrated that some IDH episodes were predictable using the variation in the PPG baseline with respiration as a surrogate for low blood volume. Additionally, the area under the curve of the PPG waveform can be used as a surrogate for cardiac output and peripheral vascular tone, resulting in a reasonable predictor for potentially critical changes in blood pressure during dialysis. Individually, the novel metrics described here are limited in their identification of IDH in all patients affected, but in combination they may be used to develop a multi-parameter predictive model. The relative merits of personalised versus population-based models are explored and a conclusion is drawn that personalised multi-parameter data fusion modelling for haemodialysis patients would be an important area for future work.

Background: Minor procedures in surgery and urology such as groin hernia and hydrocele repair, as well as prostate biopsies are very frequently done in routine practice. Complications and insufficient outcomes thus affecting many patients and the cumulative effect of this are of major importance in a population perspective. Aim: To explore complications and outcomes of surgical or diagnostic procedures and possible risk factors or predictors for adverse effects. Methods: By using both national quality and administrative registers, and by complementing registers with patient reported outcome measures, examine outcomes such as complications, persistent pain and recurrences. Also, in the case of hydro and spermatoceles, report incidence numbers. Further, by using a randomized trial, explore minimally invasive procedure such as sclerotheraphy compared to conventional surgery in respect to cure and adverse events. Results: When comparing with the open anterior mesh repair, endoscopic technique is advantageous in respect to the patient reported outcome of persistent pain. The drawback was an increased risk of postoperative complications and reoperation for recurrence. Incidence numbers for hydro and spematocele were 100/100000 men. Aspiration (± sclerotherapy) had a significantly lower rate of complications as compared to conventional surgery. In the interim analysis of the randomized trial, comparing sclerotherapy to Lord´s procedure for hydroceles, the cure rate was similar between treatments. Definite conclusions cannot be made due to the risk of type 2 errors, and the study will thus continue. In the case of trans-rectal prostate biopsy, the rates increased every year during the study time frame, up to an approximate risk of two per cent in 2012 for hospital readmission within 30 days, without an increased mortality within 30 days. Conclusions: The open anterior mesh procedure is still the preferred method for groin hernia repair in routine surgical practice. Hydro and spermatocele surgery is associated with high rates of complications, and the indication for repair should be scrutinized. The rates of infection after prostate biopsy is increasing and methods to reduce unnecessary biopsies as well as improved prophylaxis should be investigated.

Colorectal cancer (CRC) is defined as a cancer appearing in the colon or in the rectum. In Sweden, ~ 6300 individuals were diagnosed with the disease in 2014 and ~ 2550 individuals diagnosed with CRC die each year due to their cancer. Surgery is the main treatment option of CRC and a survival rate of ~ 10 % is estimated if distant metastases have developed. It is therefore of importance to find factors that may be useful together with tumour, node, metastasis (TNM) stage to establish early CRC diagnosis, prognosis and follow-up of CRC patients. The aim of this thesis was to study the possible association of CD93, PLA2G4C, PDGF-D and inflammatory cytokines with CRC disease progression. In a prospective study approach CD93 and PLA2G4C single nucleotide polymorphisms (SNPs) were of potential importance in CRC prognosis. The T/T genotype of CD93 was associated with an increased CD93 expression in CRC tissue. Further, CRC patients carrying this genotype were associated with disseminated CRC at diagnosis and a lower recurrence-free survival after surgery. The A allele of a SNP of PLA2G4C was a stronger predictor for CRC-specific mortality than the conventional risk factors used in the clinic for selection of TNM stage II patients for adjuvant treatment. This indicates that the T/T genotype of CD93 and the A allele of PLA2G4C may be potential genetic factors related to disease severity and spread. Furthermore, they distinguish CRC patients that may benefit from a more comprehensive follow-up and adjuvant treatment. To study the putative involvement of PDGF-D in CRC the effects of PDGF-D signalling was studied in vitro. PDGF-D signalling altered the expression of genes of importance in CRC carcinogenesis and proliferation which was blocked by imatinib, a tyrosine kinase inhibitor. This indicates that PDGF-D signalling may be an important pathway in CRC progression and a potential target in CRC treatment. The analysis of various inflammatory cytokines in plasma at diagnosis showed an association between high levels and increased total- or CRC-specific mortality two years after surgery. High levels of CCL1 and CCL24 was the only cytokines strongly correlated with a worse CRC prognosis after statistical adjustments and may be of interest for further evaluation. In conclusion, this thesis presents circulating and genetic factors such as CD93, PLA2G4C, PDGF-D, CCL1 and CCL24 that may be of importance in CRC progression and may be of clinical value together with TNM stage in establishing prognosis. / Kolorektal cancer är en tumör i kolon eller rektum. I Sverige diagnosticerades år 2014 ca6300 individer med denna cancertyp och ca 2550 personer dör årligen till följd av kolorektalcancer. Operation är det huvudsakliga behandlingsalternativet för kolorektal cancer och vidfjärrmetastaser är överlevnaden < 10 %. Det är därför viktigt att hitta markörer somtillsammans med TNM-stadium kan ge tidig information om sjukdomens prognos och lämpliguppföljning av patienter. Utveckling av kolorektal cancer sker genom ackumulering av genetiska mutationer ochepigenetisk nedreglering av tumörsuppressorgener. Därutöver spelar interaktionen mellantumören och dess närmaste omgivning, innehållande tillväxt- och inflammatoriska faktorer,en viktig roll i tumörens utveckling och metastasering. Syftet med avhandlingen var att studera associationen mellan CD93, PLA2G4C, PDGF-D samtinflammatoriska cytokiner och kolorektal cancer progression. En prospektiv studie visade att CD93 och PLA2G4C SNP var potentiellt viktiga förbedömningav kolorektal cancer prognos. T/T genotypen av SNP rs2749817 i CD93 var associerad medhögre uttryck av CD93 i kolorektal cancer vävnad, främst bland patienter i stadium IV.Därutöver observerades fler återfall efter operation hos patienter med T/T genotypen. Aallelen hos PLA2G4C SNP rs1549637 är en möjligtvis bättre markör för cancerspecifiköverlevnad vid stadium II än faktorer som idag används för att selektera patienter tilladjuvant behandling. Sammantaget antyder detta att T/T genotypen av CD93 och A allelenav PLA2G4C kan vara genetiska markörer relaterade till allvarlig tumörsjukdom ochspridning. Därutöver kan de eventuellt selektera patienter som kräver tätare uppföljning ochadjuvant behandling. För att studera den förmodade inblandningen av PDGF-D i kolorektal cancer undersöktesdess effekt på PDGF-D signalering in vitro. PDGF-D signaleringen förändradegenexpressionen av gener involverade i tumörutveckling och spridning, vilken kundeblockeras av tyrosinkinashämmaren imatinib. Det antyder att PDGF-D signalering kan vara enviktig faktor vid kolorektal cancer progression och ett potentiellt mål för behandling. Analysen av ett flertal inflammatoriska cytokiner visade en korrelation mellan högacytokinnivåer och ökad cancerspecifik och total dödlighet två år efter operation. Höga CCL1och CCL24 nivåer var de enda faktorerna som förblev signifikant associerade medcancerspecifik mortalitet vid fördjupad statistisk analys och bör studeras vidare. Sammanfattningsvis presenterar denna avhandling cirkulerande och genetiska faktorersåsom CD93, PLA2G4C, PDGF-D, CCL1 and CCL24 som eventuellt är viktiga vid bedömning avkolorektal cancer progression tillsammans med TNM stadium.

Hypoxia regulates many hundreds of genes with important roles in ischemic and neoplastic disorders. Central to this response are the hypoxia inducible transcription factors (HIF). This work aimed to better understand the direct transcriptional response to HIF by mapping HIF-binding sites across the genome using chromatin immunoprecipitation coupled to high-throughput sequencing (ChIP-seq). ChIP-seq for HIF in MCF-7 breast cancer cells under hypoxic conditions revealed more than 400 high-stringency HIF-binding sites genome-wide. Each member of the HIF heterodimer was present with near complete concordance. Binding of the two principle isoforms revealed a high degree of overlap with no differences in the DNA-binding motif. HIF-binding was associated with upregulation, but not downregulation of genes indicating that it functions as a transcriptional activator but not as a repressor. HIF-binding occurred preferentially at gene promoters, but was also present at promoter-distant sites, which were also associated with gene regulation, implicating long-range interactions in hypoxic gene activation. HIF-binding was associated with markers of open chromatin and active enhancers that were present in normoxia, indicating that HIF-binding sites are already “prepared” to bind HIF before the hypoxic stimulus. Analysis of normoxic and hypoxic RNA pol2 and H3K4me3 signals revealed distinctive hypoxia-inducible changes unique to HIF-binding genes. Comparable numbers of HIF-binding sites were observed in a second cell line (von Hippel-Lindau defective 786-O renal cancer cells) as in MCF-7 breast cancer cells, although approximately 65% were unique to 786-O cells. These unique sites were more frequently promoter-distant. Correlation with expression analyses from renal tumours indicated that many HIF-binding genes were upregulated in renal cancer. One such RCC unique promoter-distant HIF-binding site was identified at an intergenic locus on chromosome 11q13.3 that has been associated with renal cancer in Genome-Wide Association Studies. The HIF-binding site was in high linkage disequilibrium with the disease associated SNP and had the epigenetic hallmarks of an enhancer. Analysis of pan-genomic expression analyses identified the cell-cycle regulator cyclin D1 as highly HIF-regulated, and a physical association between the HIF-binding site and the CCND1 promoter could be determined. Furthermore, in a renal cancer cell line heterozygous at this locus, the RCC-protective allele disrupted HIF-binding leading to an allelic imbalance in cyclin D1 expression.

Chronic kidney disease (CKD), defined as a decrease in renal function, is a global issue. The treatment of CKD and its comorbidities imparts a costly burden on the American healthcare system, therefore the need for therapeutics that prevent the progression of chronic kidney disease is urgent. Microarray studies have shown that the serine protease inhibitor clade A member 3 (SERPINA3) is transcriptionally upregulated in kidney injury. SERPINA3 is an extracellular protease inhibitor that maintains the homeostasis of extracellular matrix proteins. Our lab hypothesizes that SERPINA3 might not only be a transcriptional biomarker for kidney injury, but the SERPINA3 protein might act as a key upstream regulator in the advancement of renal inflammation and fibrosis. Our research characterizes the expression patterns of SERPINA3 in models of acute and chronic kidney injury through immunoblotting and immunohistochemistry. Our unilateral ureteral obstruction (UUO) model of chronic renal injury displays significant glomerular localization of SERPINA3. The adenine diet model of chronic kidney injury and the renal ischemic reperfusion injury (RIRI) model of acute kidney injury both display tubular upregulation of SERPINA3. The DOCA-salt hypertension model of chronic kidney injury was imposed on two strains of mice, C57BL/6 and 129/sv, both of which display tubular and glomerular upregulation of SERPINA3. However, the C57BL/6 strain, which is known for its resistance to glomerular sclerosis, displays higher renal localization of SERPINA3 when exposed to DOCA-salt hypertension, than does the 129/sv strain. In conclusion, our data suggests that SERPINA3 protein is upregulated in both acute and chronic kidney injury. The role of SERPINA3 in these models remains unknown, however, our lab theorizes that SERPINA3 protein may be renoprotective in certain instances of kidney injury. Functional assays must be performed to elucidate the role of SERPINA3 in these models of kidney injury. Characterizing the function of SERPINA3 in chronic and acute kidney injury might aid in the development of novel therapeutics to prevent the advancement of CKD.

Sympathetic nerves are important for renal physiology and sepsis pathophysiology. A recent study showed sprouting of sympathetic nerves in spleen of septic mice. This study was done to test if renal sprouting of sympathetic nerves also happens and to investigate renal morphology in septic mice. Cecal ligation and puncture (CLP) was used to induce sepsis and kidneys were removed for evaluation. Bowman’s space was diminished with cortical bubble cells present suggestive of acute renal pathology, however, renal function was unchanged. Acute sepsis did not affect either renal sympathetic innervation or non-neuronal cholinergic cells. Mouse kidneys had more epinephrine (EPI) than norepinephrine (NE) in both groups. This is most likely due to uptake of epinephrine by renal sympathetic nerves and may have no correlation with sepsis. In conclusion, septic mice showed minor renal pathology and no evidence of acute sympathetic nerve sprouting. Further studies are needed to understand the mechanism and consequences of elevated EPI in mice kidney.