Epidemiology and Clinical Characteristics of Chronic Kidney Disease in Bihar: A Cross-Sectional Study From a Single Center.

Background Chronic kidney disease (CKD) is a major public health concern globally, often co-occurring with type 2 diabetes (T2D), hypertension (HTN), and cardiovascular disorders (CVD), which complicate its management and exacerbate outcomes. This study aims to investigate the epidemiological and clinical characteristics of CKD in Bihar, a region often underrepresented in national data. Methods This cross-sectional observational study was conducted at the Department of Nephrology, Indira Gandhi Institute of Medical Sciences (IGIMS) in Patna, Bihar, India. A total of 2,534 adult patients of both sexes who consented to participate were included. We collected demographic and clinical data, calculated the estimated glomerular filtration rate using the CKD-Epidemiology (CKD-EPI) Collaboration creatinine equation, and classified CKD stages. Statistical analyses were performed using IBM SPSS Statistics for Windows, version 29.0.2.0 (IBM Corp., Armonk, NY). Result The majority of the study population was male (66.5%), with a significant number residing in rural areas (76.8%). The prevalent causes of CKD included HTN (41.2%), chronic tubulointerstitial nephritis (31.8%), and T2D (23.2%). Approximately one-third of patients were in the early stages (Stages 1 and 2) of CKD. A high prevalence of anemia was noted across all stages, increasing significantly with glomerular filtration rate (GFR) reduction. Treatment analysis showed suboptimal use of angiotensin-converting enzyme inhibitors (ACEi) or angiotensin receptor blockers (ARBs) and other standard treatments like diuretics and statins, especially among T2D patients. Conclusion Chronic kidney disease in Bihar affects predominantly young males and is associated with significant rural prevalence and comorbidities like T2D, HTN, and CVD. Our results highlight the need for improved management practices, especially in the use of ACEi/ARBs and erythropoiesis-stimulating agents, to slow GFR reduction. Further multicentric, community-based studies are recommended to provide a more comprehensive understanding of CKD in Bihar.

A Series of Eight Cases of Pigment Nephropathy: An Obscured Aspect of Acute Kidney Injury.

Pigment-induced acute kidney injury (AKI) is an important and preventable complication of rhabdomyolysis or hemolysis. It is characterized by the release of free heme pigment (myoglobin or hemoglobin) in the circulation, leading to direct injury of the proximal tubule and distal tubule obstruction by pigment cast. We are reporting eight cases of pigment-induced AKI, including six cases of myoglobin cast nephropathy and two cases of hemoglobin cast nephropathy. The causes of rhabdomyolysis were strenuous exercise, infection/febrile illness, and drug-induced neuroleptic malignant syndrome. Paroxysmal nocturnal hemoglobinuria and anti-tuberculosis treatment (rifampicin and isoniazid) had led to hemoglobin cast nephropathy each in one case. Seven cases had severe renal failure requiring dialysis. Short-term renal outcome was favorable. However, long-term follow-up is necessary to determine whether pigment-induced AKI has delayed sequelae. Therefore, clinicians should consider rhabdomyolysis or hemolysis as potential hidden causes of AKI in diverse clinical conditions, especially those of non-traumatic origin, to achieve an accurate diagnosis.

Clinical Profile and Short-Term Outcomes of Acute Kidney Injury in Elderly Patients in a Tertiary Care Center.

Background Acute kidney injury (AKI) is a common and serious condition affecting elderly patients. Despite its significance, comprehensive research focusing specifically on the clinical profile and short-term outcomes of AKI in this vulnerable population is lacking. Objective This study aimed to evaluate the clinical profile and short-term outcomes of AKI in elderly patients admitted to a tertiary care center. Methods A prospective observational study was conducted from December 2023 to March 2024, involving 75 elderly patients (aged ≥65 years) diagnosed with AKI. Baseline demographic data, clinical profiles, laboratory investigations, mortality rate among elderly patients diagnosed with AKI within 30 days of diagnosis, and short-term outcomes were recorded and analyzed. Descriptive statistics and appropriate statistical tests were used for the data analysis. Results The study cohort had a mean age of 72.6 years. Hypertension was present in 55 patients (73.3%), and diabetes mellitus was observed in 30 patients (40.0%). Prerenal causes of AKI were identified in 40 patients (53.3%), while acute tubular necrosis was found in 25 patients (33.3%). Stage 2 AKI was the most common, affecting 35 patients (46.7%). Out of the 75 patients, 15 patients (20.0%) succumbed to AKI within the study period. Deceased patients had longer hospital stays, with a median of 16 days compared to 10 days for survivors. ICU admission was required for 13 of the deceased patients (86.7%), compared to 32 of the surviving patients (53.3%). The need for renal replacement therapy was higher among the deceased patients, with 11 out of 15 patients (73.3%) requiring it, compared to 19 out of 60 surviving patients (31.7%). Renal function recovery was notably lower in the deceased patients. Conclusion AKI in elderly patients was associated with significant morbidity and mortality, highlighting the need for early recognition, appropriate management, and preventive strategies. A comprehensive evaluation of the clinical profile and short-term outcomes of AKI in the elderly population provides valuable insights for optimizing patient care and improving outcomes.

The Clinicopathological Correlation and Outcome of Glomerulonephritis With Crescent: A Single-Center Study.

BACKGROUND: There is a lack of standardized treatments for patients with less than 50% crescents observed in their renal biopsies. This study aimed to analyze the crescent percentage, clinicopathological characteristics, and renal prognosis of glomerulonephritis (GN) cases with at least one crescentic lesion. MATERIALS AND METHODS: This retrospective cohort study was conducted at the Indira Gandhi Institute of Medical Sciences, Patna, from January 2016 to December 2020. Consecutive patients (aged between 18 and 65 years) with renal biopsy findings suggestive of GN and at least one crescent were included in the study. Demographic details and clinical presentation were collected from the medical records. RESULTS: A total of 145 patients were included. The mean (standard deviation (SD)) age was 33.06 (11.739) years. Hemoptysis was significantly higher in the ≥50% crescent group (P=0.011). Rapidly progressive glomerulonephritis (RPGN) was significantly higher in the ≥50% crescent group (P<0.001). There was a significant difference observed in mean creatinine (P=0.001), mean crescents (P<0.001), and mean urine polymerase chain reaction (PCR) (P=0.031). Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis was significantly higher in the ≥50% crescent group (P<0.001). Complete remission decreased as crescents increased. In GN with crescent, the presence of fibrous crescents (≥50%) is associated with a higher rate of treatment resistance (100%) compared to fibrocellular (58.33%) and cellular crescents (6.25%). In the ≥50% crescent group, death was significantly higher in patients with fibrous crescent age (57.14%). CONCLUSION: Crescent percentage and crescent age were found to be significantly related to greater risk of renal failure and resistance to treatment.

Clinical Profile and Outcomes of Nephrotic Syndrome With Acute Kidney Injury in Adults: A Prospective Cohort Study.

BACKGROUND: Nephrotic syndrome (NS) is characterized by various clinicopathological conditions like proteinuria, hypoalbuminemia, and anasarca. Patients with NS are prone to experience associated problems like acute kidney injury (AKI). The present study aimed to investigate the clinical profile and outcomes of NS with AKI in adults. MATERIALS AND METHODS: This prospective, observational study was conducted over a period of one year. Adult patients with NS diagnosed with AKI were enrolled in the study. Data were collected at baseline and patients were followed up for at least three months. RESULTS: A total of 60 patients were enrolled. The majority of the patients (78.3%) were aged between 18 and 30 years. Anemia was observed among 96.7% of the patients. A significant improvement was observed in the mean levels of proteinuria (5.80 vs. 1.70 gm/dL; P < 0.001), total cholesterol (270.00 mg/dL vs. 160.00 mg/dL), serum creatinine (2.18 mg/dL vs. 1.68 mg/dL; P < 0.001), and serum albumin (1.86 gm/dL vs. 3.29 gm/dL; P < 0.001) at baseline to three months. Pre-renal AKI was diagnosed in 95% of patients. According to histological classification, 19 patients had minimal change disease, whereas focal segmental glomerulosclerosis was observed in 23 patients. It was observed that 96.7% of the patients did not necessitate the need for renal replacement therapy. CONCLUSION: The present study successfully examined the clinical profile and outcomes of adults with NS and AKI. The findings provide valuable insights into the characteristics and prognosis of this patient population, contributing to a better understanding of NS with AKI in adults.

Delineation of breast cancer cell hierarchy identifies the subset responsible for dormancy.

The bone marrow (BM) is a major organ of breast cancer (BC) dormancy and a common source of BC resurgence. Gap junctional intercellular communication (GJIC) between BC cells (BCCs) and BM stroma facilitates dormancy. This study reports on a hierarchy of BCCs with the most immature subset (Oct4(hi)/CD44(hi/med)/CD24(-/+)) demonstrating chemoresistance, dormancy, and stem cell properties: self-renewal, serial passaging ability, cycling quiescence, long doubling time, asymmetric division, high metastatic and invasive capability. In vitro and in vivo studies indicated that this subset was responsible for GJIC with BM stroma. Similar BCCs were detected in the blood of patients despite aggressive treatment and in a patient with a relatively large tumor but no lymph node involvement. In brief, these findings identified a novel BCC subset with stem cell properties, with preference for dormancy and in the circulation of patients. The findings establish a working cellular hierarchy of BCCs based on phenotype and functions.

Gap junction-mediated import of microRNA from bone marrow stromal cells can elicit cell cycle quiescence in breast cancer cells.

Bone marrow (BM) metastasis of breast cancer (BC) can recur even decades after initial diagnosis and treatment, implying the long-term survival of disseminated cancer cells in a dormant state. Here we investigated the role of microRNAs (miRNA) transmitted from BM stroma to BC cells via gap junctions and exosomes in tumor cell quiescence. MDA-MB-231 and T47D BC cells arrest in G(0) phase of the cell cycle when cocultured with BM stroma. Analyses of miRNA expression profiles identified numerous miRNAs implicated in cell proliferation including miR-127, -197, -222, and -223 targeting CXCL12. Subsequently, we showed that these CXCL12-specific miRNAs are transported from BM stroma to BC cells via gap junctions, leading to reduced CXCL12 levels and decreased proliferation. Stroma-derived exosomes containing miRNAs also contributed to BC cell quiescence, although to a lesser degree than miRNAs transmitted via gap junctions. This study shows that the transfer of miRNAs from BM stroma to BC cells might play a role in the dormancy of BM metastases.

RE-1-silencing transcription factor shows tumor-suppressor functions and negatively regulates the oncogenic TAC1 in breast cancer cells.

Breast cancer remains the most prevalent cancer among women in the United States. Substance P, a peptide derived from the TAC1 gene, mediates oncogenic properties in breast and other cancers. TAC1 expression facilitates the entry of breast cancer cells into bone marrow. The transcriptional repressor element 1-silencing transcription factor (REST) has been implicated in both oncogenic and tumor-suppressor functions. REST binds to the 5' untranslated region of the TAC1 promoter and suppresses its expression. This study investigated a role for REST in TAC1 induction in breast cancer. Western blots and real-time PCR indicated that REST expression in breast cancer cells was inversely proportional to the cells' aggressiveness, for both cell lines and primary breast cancer cells. REST knockdown in low-metastatic T47D cells and nontumorigenic MCF12A cells resulted in increases in TAC1 induction, proliferation, and migration. These parameters were negatively affected by ectopic expression of REST in highly aggressive MDA-MB-231 cells. Together, these findings show a central role for REST in the oncogenic function of TAC1 and suggest a tumor-suppressor role for REST in breast cancer.

Mesenchymal stem cells in early entry of breast cancer into bone marrow.

BACKGROUND: An understanding of BC cell (BCC) entry into bone marrow (BM) at low tumor burden is limited when compared to highly metastatic events during heavy tumor burden. BCCs can achieve quiescence, without interfering with hematopoiesis. This occurs partly through the generation of gap junctions with BM stroma, located close to the endosteum. These events are partly mediated by the evolutionary conserved gene, Tac1. METHODOLOGY/PRINCIPAL FINDINGS: This study focuses on the role of mesenchymal stem cells (MSCs), Tac1, SDF-1 and CXCR4 in BCC entry into BM. The model is established in studies with low numbers of tumor cells, and focuses on cancer cells with low metastatic and invasion potential. This allowed us to recapitulate early event, and to study cancer cells with low invasive potential, even when they are part of larger numbers of highly metastatic cells. A novel migration assay showed a facilitating role of MSCs in BCC migration across BM endothelial cells. siRNA and ectopic expression studies showed a central role for Tac1 and secondary roles for SDF-1alpha and CXCR4. We also observed differences in the mechanisms between low invasive and highly metastatic cells. The in vitro studies were verified in xenogeneic mouse models that showed a preference for low invasive BCCs to BM, but comparable movement to lung and BM by highly metastatic BCCs. The expressions of Tac1 and production of SDF-1alpha were verified in primary BCCs from paired samples of BM aspirates and peripheral blood. CONCLUSIONS/SIGNIFICANCE: MSC facilitate BCC entry into BM, partly through Tac1-mediated regulation of SDF-1alpha and CXCR4. We propose a particular population of BCC with preference for BM could be isolated for characterization. This population might be the subset that enter BM at an early time period, and could be responsible for cancer resurgence and resistance to current therapies.

Role of human HGFIN/nmb in breast cancer.

INTRODUCTION: HGFIN, previously identified as nmb, and its homolog osteoactivin are single transmembrane proteins that are expressed in differentiated immune cells. These proteins exhibit properties that could potentiate tumorigenesis or decrease invasiveness. These seemingly opposing roles of HGFIN suggest that this protein might be central to malignancies and might also behave as a tumor suppressor. Consistent with the reported roles for HGFIN is the fact that this gene is regulated by p53 through multiple binding sites in the 5' flanking region, and is expressed in osteoblasts. METHODS: This study used siRNA to knock-out HGFIN in non-tumorigenic breast cells and ectopically expressed HGFIN in breast cancer cells. In addition, in situ hybridization studies analyzed primary breast tissues from archived breast surgeries. Reporter gene assays studied the untranslated exon 1 of HGFIN. RESULTS: HGFIN expression led to reduced cell growth of breast cancer cells and reduced migration. At the molecular level, reporter gene analyses determined the untranslated exon 1 to be a negative regulator of the upstream enhancing effect. Ectopic expression of wild-type p53 in breast cancer cells that expressed endogenous mutant p53 resulted in increased HGFIN reporter gene activities. CONCLUSION: As the majority of cancer cells have mutations in p53, further studies on the relationship between p53 and HGFIN expression, and its role in tumor genesis and bone invasion, might uncover novel therapy targets for breast and other cancers. The results show a central role for p53 in HGFIN expression, which appears to determine the behavior of the cancer cells.

Nuclear factor-kappaB is central to the expression of truncated neurokinin-1 receptor in breast cancer: implication for breast cancer cell quiescence within bone marrow stroma.

Breast cancer is a leading cause of mortality among women in the United States. Tac1 and neurokinin-1 (NK1) are involved in autocrine stimulation of breast cancer cells (BCCs). The single NK1 gene produces full-length (NK1-FL) and truncated (NK1-Tr) forms. NK1-Tr mediates malignancy in breast cells. We now report a critical role for nuclear factor-kappaB (NF-kappaB) in the expression of NK1-Tr, but not NK1-FL, in human BCCs. By Western and Northern blot analyses, NK1-FL and NK1-Tr were coexpressed in BCCs but were undetectable in nontumorigenic cells. Loss of repressive activity within the 5' flanking region of the NK1 partly accounts for constitutive expression of NK1 in BCCs but could not account for the presence of NK1-Tr. Transient transfections with dominant-negative and wild-type IkappaB show that activation of NF-kappaB is required for the expression of NK1-Tr. Tac1 gene was linked to the generation of NK1-Tr because its overexpression in BCCs led to the production of multiple cytokines that can activate NF-kappaB to mediate NK1-Tr expression. Studies with Tac1 knockdown BCCs and Tac1-expressing nontumorigenic breast cells verified a role for NF-kappaB in the expression of NK1-Tr. The quiescent phenotype of BCCs on contact with bone marrow stroma was partly explained by decreased NF-kappaB activation and undetectable NK1-Tr. In summary, this study shows a role for NF-kappaB in the expression of NK1-Tr in BCCs, which seems to be reversed by bone marrow stromal cells.

SDF-1alpha regulation in breast cancer cells contacting bone marrow stroma is critical for normal hematopoiesis.

Breast cancer cells (BCCs) show preference for the bone marrow (BM). An animal model showed 2 populations of BCCs in the BM with regard to their cycling states. An in vitro model of early BC entry into BM showed normal hematopoiesis. Here, we show a critical role for BCC-derived SDF-1alpha in hematopoietic regulation. The studies used a coculture of BM stroma and BCCs (cell lines and stage II BCCs). Northern blots and enzyme-linked immunosorbent assay (ELISA) showed gradual decreases in SDF-1alpha production in BCCs as they contact BM stroma, indicating partial microenvironmental effects caused by stroma on the BCCs. SDF-1 knock-down BCCs and increased exogenous SDF-1alpha prevented contact inhibition between BCCs and BM stroma. Contact inhibition was restored with low SDF-1alpha levels. Long-term culture-initiating assays with CD34(+)/CD38(-)/Lin(-) showed normal hematopoiesis provided that SDF-1alpha levels were reduced in BCCs. Gap junctions (connexin-43 [CX-43]) were formed between BCCs and BM stroma, with concomitant interaction between CD34(+)/CD38(-)/Lin(-) and BM stroma but not with the neighboring BCCs. In summary, SDF-1alpha levels are reduced in BCCs that contact BM stroma. The low levels of SDF-1alpha in BCCs regulate interactions between BM stroma and hematopoietic progenitors, consequently facilitating normal hematopoiesis.

Transformation of breast cells by truncated neurokinin-1 receptor is secondary to activation by preprotachykinin-A peptides.

Breast cancer remains the cancer with the highest mortality among women in the United States. Peptides derived from the oncogenic Tac1 gene (full transcript: betaPPT-A) stimulate the proliferation of breast cancer cells (BCCs) via seven-transmembrane G protein-coupled neurokinin 1 (NK1) and NK2 receptors. The NK1 gene could generate full-length (NK1-FL) and truncated (NK1-Tr) transcripts. NK1-Tr lacks 100 residues in their cytoplasmic end, could couple to G proteins, and shows reduced efficiency with respect to internalization and desensitization. This study reports on a role of NK1-Tr in the transformation of nontumorigenic breast cells, and investigates whether Tac1 expression is linked to the generation of NK1-Tr. Western blots and Northern analyses showed coexpressions of NK1-Tr and NK1-FL in BCCs (cell lines and primary cells from patients with different stages of breast cancer). Stable transfections of betaPPT-A or NK1-Tr expression vectors in nontumorigenic cells showed each induces the expression of the other, consequently resulting in a transformed phenotype. Analyses with microarrays indicate similar patterns of cytokine production by NK1-Tr transfectants and BCCs, but not NK1-FL transfectants. These observations indicate tumor-promoting properties by NK1-Tr, but not NK1-FL. Overall, the oncogenic property of Tac1 in breast cells involves concomitant expression of NK1-Tr and vice versa, consequently leading to the production of cytokines with growth promoting functions.

An in vitro method to select malignant cells from surgical biopsies of breast cancer patients.

To date, breast cancer (BC) research is mainly studied with cell lines. These cells were passaged multiple times, acquiring phenotypes, additional mutations and epigenetic changes. These changes make the passaged cell lines different from the original malignancy. Thus cell lines, although useful as models could be improved with additional studies with primary BC. It is difficult to obtain malignant cells from breast tissues without contamination from surrounding healthy cells. Selection and expansion of malignant cells from surgical tissues have proved to be daunting tasks. This study describes a reliable and reproducible method for isolating and expanding malignant cells from surgical breast tissues. The method uses co-cultures with BM stroma to select for the cancer cells while the healthy cells undergo rapid cell death. Studies are described to show the cloning efficiencies and sensitivity of the method using surgical samples of varying sizes, different stages of BC, and samples from needle biopsies.

Bone marrow stroma influences transforming growth factor-beta production in breast cancer cells to regulate c-myc activation of the preprotachykinin-I gene in breast cancer cells.

Breast cancer cells (BCCs) have preference for the bone marrow (BM). This study used an in vitro coculture of BCCs and BM stroma to represent a model of early breast cancer metastasis to the BM. The overarching hypothesis states that once BCCs are in the BM, microenvironmental factors induce changes in the expression of genes for cytokines and preprotachykinin-I (PPT-I) in both BCCs and stromal cells. Consequently, the expression of both PPT-I and cytokines are altered to facilitate BCC integration within BM stroma. Cytokine and transcription factor arrays strongly suggested that transforming growth factor-beta (TGF-beta) and c-myc regulate the expression of PPT-I so as to facilitate BCC integration among stroma. Northern analyses and TGF-beta bioassays showed that stromal cells and BCCs influence the level of PPT-I and TGF-beta in each other. In cocultures, PPT-I and TGF-beta expressions were significantly (P < 0.05) increased and decreased, respectively. TGF-beta and PPT-I were undetectable in separate stromal cultures but were expressed as cocultures. Two consensus sequences for c-myc in the 5' flanking region of the PPT-I gene were shown to be functional using gel shift and reporter gene assays. Mutagenesis of c-myc sites, neutralization studies with anti-TGF-beta, and transient tranfections all showed that c-myc is required for TGF-beta-mediated induction of PPT-I in BCCs. TGF-beta was less efficient as a mediator of BCC integration within stroma for c-myc-BCCs. Because the model used in this study represents BCC integration within BM stroma, these studies suggest that TGF-beta is important to the regulation of PPT-I in the early events of bone invasion by BCCs.

Facilitating role of preprotachykinin-I gene in the integration of breast cancer cells within the stromal compartment of the bone marrow: a model of early cancer progression.

Despite early detection of breast cancer, patients' survival may be compromised if the breast cancer cells (BCCs) enter the bone marrow (BM). It is highly probable that BCCs enter the BM long before clinical detection. An in vitro coculture model with BM stroma and BCCs (cell lines; primary cells from stage III BC, n = 7, and stage M0, n = 3) mimicked early entry of BCCs into the BM. In coculture, BCCs exhibit contact inhibition and do not require otherwise needed growth supplements. Stromal growth rate was increased 2-fold in coculture. The inclusion of BCCs in stromal support of long-term culture-initiating cell assay frequencies show no difference (38 +/- 3 versus 36 +/- 6). Nontumorigenic breast cells (patients and cell lines) did not survive in coculture, suggesting that the model could select for malignant population in surgical breast tissues. Cocultures were able to select cells with 73 +/- 7% cloning efficiencies and with the ability to form cocultures with BM stroma. Preprotachykinin-I (PPT-I), a gene that is conserved by evolution, facilitates BCC integration as part of the stromal compartment. This was deduced as follows: (a) nontumorigenic breast cells (n = 4) genetically engineered to express PPT-I and led to anchorage-independent growth, foci formation, and formation of cocultures; and (b) suppression of PPT-I in BCCs (n = 5) with pPMSKH1-PPT-I small interfering RNA reverted the cells to nontumorigenic phenotypes and was undetectable in the BM nude mice. The evidence supports that the PPT-I gene facilitates the integration of BCCs in the stromal compartment during a period before clinical detection, without disrupting hematopoietic activity.