Mini-PCNL - a boon for CKD patients with nephrolithiasis.

Purpose: The prevalence of urinary stone disease in ESRD is 3.2%, leading to renal damage due to obstructive uropathy, infection, and frequent surgical intervention. PCNL, the gold standard for complex renal stone disease, has evolved with smaller access sheaths (14-20 F), improved optics, and fluoroscopic equipments. This study aims to assess the safety and feasibility of mini- PCNL in CKD patients with respect to postoperative outcome and its effect on stage of CKD. Patients and methods: This prospective study was conducted in the Department of Urology, Sher-I-Kashmir Institute of Medical Sciences, from January 2022 to October 2022. This study included adult patients with nephrolithiasis at CKD stage 3 or higher. The primary objective of this research was to assess the impact of mini-PCNL on renal function, specifically measuring changes in estimated glomerular filtration rate (eGFR) from baseline to a 6-month follow-up. The secondary objective was to evaluate the feasibility of mini-PCNL in CKD patients in terms of complications, stone clearance rate, and duration of hospital stay. Four variable Modification of Diet in Renal Diseases(MDRD) equation was used to calculate the estimated GFR(eGFR) of each patient and NKF/KDOQI classification system to classify the stage of CKD. Results: A total of 46 patients were included in the study. We found that for management of nephrolithiasis in CKD patients, mini-PCNL leads to significant improvement in eGFR at 6 months follow-up (mean difference = 14.25 ml/min/1.73 m2; p-value <0.01) with high stone-free rates (89.5%). The complication rates were comparable to standard PCNL. Conclusions: mini-PCNL is a bonanza for management of CKD patients with nephrolithiasis.

Characterization of palmitic acid toxicity induced insulin resistance in HepG2 cells.

BACKGROUND: An etiology of palmitic acid (PA) induced insulin resistance (IR) is complex for which two mechanisms are proposed namely ROS induced JNK activation and lipid induced protein kinase-C (PKCε) activation. However, whether these mechanisms act alone or in consortium is not clear. METHODS AND RESULTS: In this study, we have characterized PA induced IR in liver cells. These cells were treated with different concentrations of PA for either 8 or 16 h. Insulin responsiveness of cells treated with PA for 8 h was found to be same as that of control. However, cells treated with PA for 16 h, showed increased glucose output both in the presence and in absence of insulin only at higher concentrations, indicating development of IR. In these, both JNK and PKCε were activated in response to increased ROS and lipid accumulation, respectively. Activated JNK and PKCε phosphorylated IRS1 at Ser-307 resulting in inhibition of AKT which in turn inactivated GSK3ß, leading to reduced glycogen synthase activity. Inhibition of AKT also reduced insulin suppression of hepatic gluconeogenesis by activating Forkhead box protein O1 (FOXO1) and increased expression of the gluconeogenic enzymes and their transcription factors. CONCLUSION: Thus, our data clearly demonstrate that both these mechanisms work simultaneously and more importantly, identified a threshold of HepG2 cells, which when crossed led to the pathological state of IR in response to PA.

Incidence of Ureteric strictures Following Ureteroscopic Laser Lithotripsy: Holmium:YAG Versus Thulium Fiber Laser.

OBJECTIVE: We aimed to compare the incidence of ureteric strictures between holmium:yttrium aluminum garnet and thulium fiber laser following ureteroscopic laser lithotripsy. In the present era of miniaturization of endourologic armamentarium and better optics, how safe are lasers to fire inside ureter? MATERIALS AND METHODS: It is a prospective comparative study over a period of 2 years that included patients who underwent ureteroscopic laser lithotripsy for ureteric stones. Patients were randomly divided into 2 groups: group A underwent holmium:yttrium aluminum garnet laser lithotripsy and group B underwent thulium fiber laser lithotripsy. RESULTS: A total of 478 patients were analyzed after excluding patients not willing to participate and patients lost to follow-up. Two hundred forty patients underwent holmium:yttrium aluminum garnet laser lithotripsy (group A) and 238 patients underwent thulium fiber laser lithotripsy (group B). The demographic data of 2 groups were comparable. The mean age of patients in group A and group B was 36.5 ± 12.52 years and 38.62 ± 10.71 years, respectively. The mean operative time in group A and group B was 47 ± 15 and 36 ± 13 minutes, respectively, while the mean laser time in group A and group B was 13.5 ± 45 minutes and 9.25 ± 3.2 minutes, respectively. Four (1.67%) patients in group A and 11 (4.62%) patients in group B developed ureteric strictures during follow-up, and the difference was statistically significant (P <.001). The mean length of stricture was 2.67 ± 1.27 cm in group A and 4.42 ± 2.2 cm in group B, and the difference was statistically significant. CONCLUSION: Thulium fiber laser, projected as safe laser previously, has a higher incidence of ureteric strictures compared to holmium:yttrium aluminum garnet laser when used for ureteroscopic laser lithotripsy.

Improving Surgical Safety in Living Donor Renal Transplantation With Antiseptic Skin Preparation, Bladder Irrigation, Corner-Saving Vascular Anastomosis, DJ Stenting, and Extravesical Ureteroneocystostomy Modifications: A Comprehensive Approach.

Introduction The antiseptic skin preparation, bladder irrigation, corner-saving vascular anastomosis, DJ stenting, and extravesical ureteroneocystostomy (ABCDE) approach encompasses a range of modifications applied during different stages of the surgical procedure in renal transplantation. These modifications include the following: A, antiseptic skin preparation sequentially with cetrimide 3.35%, chlorhexidine scrub 4%, spirit, and povidone-iodine 10%; B, bladder irrigation with amikacin and betadine solution; C, corner-saving end-to-side vascular anastomosis; D, DJ stenting with early postoperative removal within three weeks; and E, extravesical ureteroneocystostomy using our institute's modified Lich-Gregoir technique. Methods This prospective observational study was conducted at our institution between March 2021 and May 2023. Data were collected from the patients' medical records and analyzed using Statistical Package for the Social Sciences (SPSS) (IBM SPSS Statistics, Armonk, NY, USA). Statistical tests, including t-test, Mann-Whitney test, chi-square test, and Fisher's exact test, were used for analysis. The study assessed various recipient, donor, intraoperative, and post-transplant factors, as well as surgical complications and stent-related factors. Results Out of 72 renal transplantations, 12 (16.6%) had the following surgical complications: urinary (n = 4; 5.5%), wound-related (n = 3; 4.1%), and lymphocele (n = 5; 6.9%). The most common complications were lymphocele (n = 5; 6.9%) and urinary leak (n = 4; 5.5%). Surgical complications were more common in male recipients (91.6% versus 8.3%), as well as in recipients with longer dialysis duration (24 ± 17 versus 11.0 ± 7 months) and had extended hospitalization time (16.4 ± 8.6 versus 8.0 ± 2.9 days) (p < 0.05). Wound infection correlated with longer surgeries (>300 minutes) and other complications. Lymphocele patients had higher drain output (>500 mL) on day 1 and longer hospital stays (>15 days). Urinary tract infections (UTIs) were linked to dialysis duration (>24 months), diabetes, and longer indwelling times of DJ stents and urinary catheters. Early DJ stent removal (<3 weeks) reduced UTI incidence and symptoms (p < 0.05). All complications were categorized as minor (3a or less), according to the Clavien-Dindo classification. Conclusion The modified ABCDE surgical approach in renal transplantation decreased the complications, showing favorable outcomes compared to those in the literature.

"Collaterals a savior" in superior mesenteric artery injury post radical nephrectomy and retroperitoneal lymph node dissection.

Superior Mesenteric Artery (SMA) during renal surgery is rarely reported but potentially devastating complication. It can rarely occur in patients with distorted vascular anatomy like in large left renal tumors with vascular infiltration and bulky lymphadenopathy, or in the setting of re-do surgery with extensive scarring. Failure to recognize and repair an SMA injury may result in ischemic bowel and consequently high mortality. Herein, we present a case scenario of injury to the SMA during radical nephrectomy missed intraoperatively and managed conservatively in the post-operative period in view of collateral circulation to the gut.

Comparative study of tamsulosin versus tadalafil in benign prostatic hyperplasia patients with lower urinary tract symptoms. A prospective randomized study.

Introduction: Benign prostatic hyperplasia (BPH) is one of the common causes of lower urinary tract symptoms (LUTS) in aging men. Men with LUTS have a higher incidence of erectile dysfunction (ED), and LUTS themselves represent an independent risk factor for ED, triggering a significant negative impact on quality of life. Materials and Methods: A total of 92 patients were randomly assigned to two groups. Groups I and II had 45 and 47 patients, two patients from Group I and three patients from Group II did not follow and were excluded from the study. Patients in Group I received 0.4 mg of tamsulosin and Group II patients received 5 mg tadalafil. Patients were assessed at baseline, 3 months, and at 6 months after receiving treatment. Treatment efficacy was measured by a change in Qmax, post void residual urine (PVR), International Prostate Symptom Score (IPSS), and Sexual Health Inventory for Men (SHIM) score at 3 months and 6 months. Results: Baseline parameters between the two groups were similar. Mean Qmax improved by 7 ml/s at 3 months to 9.44 ml/s at 6 months in Group I versus 4.73 ml/s at 3 months to 6.46 ml/s at 6 months in Group II (P = 0.739). Mean PVRU decreased by 35.53 ml at 3 months to 47.23 ml at 6 months in Group I versus 44.98 at 3 months to 58.28 ml at 6 months in Group II (P = 0.102). IPSS score improved by 4.24 points at 3 months to 7.22 points at 6 months in Group I versus 4 points at 3 months to 5.02 points at 6 months in Group II (P = 0.336). SHIM score improved by 0.7 points 16.2 at 3 months to 0.8 points at 6 months in Group I versus 6.12 points at 3 months to 6.3 points at 6 months (P < 0.001). Conclusion: When both groups were compared, tadalafil showed statistically similar improvements in Qmax, PVRU, and IPSS score, but statistically significant improvement was observed with tadalafil in SHIM score compared with tamsulosin in treating LUTS secondary to BPH. Our study provides evidence that once daily tadalafil 5 mg is well tolerated and can be considered for the treatment of LUTS secondary to BPH especially in patient with ED.

Prostatic rhabdomyosarcoma in the adult, A case report.

Prostate embryonal rhabdomyosarcoma (ERMS) is a common tumour in infants and children, but it is rare in adults. It is characterized by a high degree of malignancy, both local rapid growth with formation of large pelvic masses, often leading to renal failure due to bladder outlet obstruction, and systemic spread, commonly to the lungs, liver and bone. We report on a case of a stage III prostate ERMS, approached with combined-modality treatment, with the administration of 5 courses of doxorubicin, ifosfamide and 2-mercaptoethane sulfonate sodium (mesna), with planned subsequent radiotherapy to the prostatic bed (60 Gy/30 fractions).

Molecular mechanism of anti-adipogenic effect of vitexin in differentiating hMSCs.

In this study, we evaluated a detailed molecular mechanism of anti-adipogenic activity of vitexin, apigenin flavone glucoside, present in germinated fenugreek seeds, in differentiating human mesenchymal stem cells (hMSCs). The lipid content of differentiated adipocytes was estimated by ORO staining. Effect on mitotic clonal expansion was checked by cell cycle analysis. Expression of early and terminal adipocyte differentiation markers, anti- and pro-adipogenic transcription factors and signalling intermediates regulating them was evaluated at RNA and protein level. We found vitexin to be non-cytotoxic up to 20 µM at which intracellular lipid accumulation was significantly decreased. Cell cycle analysis suggested that vitexin does not affect mitotic clonal expansion. Expression of early and late differentiation markers, such as CEBPα, CEBPß, PPARγ, FABP4, perilipin, adiponectin and Glut4 was significantly reduced in the presence of vitexin. Expression of KLF4 and KLF15, positive regulators of PPARγ, was decreased, whereas that of negative regulators, namely KLF2, GATA2, miR20a, miR27a, miR27b, miR128, miR130a, miR130b, miR182 and miR548 increased with vitexin treatment. This effect was mediated by the activation of the AMP-activated protein kinase (AMPK) pathway via the activation of LepR and additionally by inhibiting ROS. Thus, our results showed that vitexin regulates the expression of PPARγ and inhibits adipogenesis of hMSCs at an early stage of differentiation.

Transvaginal subfascial synthetic sling - "A novel technique" versus trans-obturator mid-urethral sling in female stress urinary incontinence: A comparative study.

BACKGROUND: To assess the treatment outcome and overall efficacy of the novel technique of a transvaginal subfascial synthetic sling (TVSS) in comparison to the standard trans-obturator tape (TOT-O) mid-urethral sling for female stress urinary incontinence (SUI). MATERIALS AND METHODS: The study included 206 female SUI patients managed at our institution between March 2015 and December 2019. The patients were randomly distributed into 2 comparable groups (Group A as TOT-O and Group B as TVSS) with respect to age, trouble due to SUI assessed with respect to degree of incontinence (number of episodes and diapers used per day), and body mass index (≤40 kg/m2), with 100 patients in the TOT-O group as group A and 106 patients in TVSS as group B. Preoperative variables related to the number of incontinent episodes and diapers usage were equal in both groups. The procedure was done under spinal anesthesia and results were assessed in terms of improvement in SUI as the primary outcome, any sexual dysfunction, complications, and overall satisfaction as secondary outcomes. RESULTS: Symptomatic improvement after the procedure was seen in all patients with complete resolution of symptoms in 91 patients (91.%) in the TOT-O group (group A) versus 96 patients (90.56%) in the TVSS (group B). Postoperative complications included urinary retention in 6% versus 5.6%, increased day time urinary frequency in 8% versus 6.6%, urge incontinence in 4% versus 2.8%, and groin/thigh pain in 12% versus 0.9%, respectively. Mesh incision was done in 1 (1%) versus none (0%), and local mesh excision for mesh erosion in 2 (2%) versus 1 (0.9%) at 3 months after the procedure in the TOT-O group and the TVSS group, respectively. The p value and Chi-Square test with respect to the clinical profile and satisfaction with respect to complete resolution of symptoms was calculated using Open EPI software which were insignificant. Sexual function (SF) was assessed using the Brief Index of SF for Women questionnaire. All the patients were satisfied with respect to SF at 6 months of follow-up. CONCLUSIONS: The TVSS as a novel technique for female SUI is less invasive, simpler to learn, with less postoperative pain with resolution of SUI, rapid recovery, and good personal satisfaction compared to the standard TOT-O in procedure.

Outcome of laparoscopic nephron sparing surgery using a Satinsky clamp for hilar control: a trusted tool (SKIMS experience).

BACKGROUND: Nephron sparing surgery is a well-established surgical procedure for patients with small/bilateral renal masses. During the procedure, hilar control can be achieved by using bulldog clamps individually on the renal vessels, the renal artery alone without clamping the vein, or a laparoscopic Satinsky clamp for en bloc hilar clamping. In our series, we described the outcome of laparoscopic nephron sparing surgery using a Satinsky clamp for hilar control. MATERIALS AND METHODS: All eligible cases with confirmed diagnosis of a renal mass were advised of nephron sparing surgery. The short-term outcomes were evaluated by warm ischemia time (using a Satinsky clamp especially when CT renal angiography was not available), average blood loss, and length of postoperative hospital stay. The oncological outcome was evaluated by noting the surgical margins of histopathological specimen, local recurrence, and distant metastasis. RESULTS: Of 30 cases 20 were male. The mean age was 54.25 years. On preoperative evaluation, 24 cases were T1a stage and the rest were 6 T1b stage. Four tumors were located in the upper pole, 4 in the posterior midpole, and 22 in the lower pole. Twenty-six patients had a low complexity score on RENAL scoring (ie, 4-6) and 4 patients a medium complexity score (ie, 7-9). Three patients were converted to open partial nephrectomy because of technical difficulty in intracorporeal suturing and difficulty in achieving hemostasis. Among these 3 patients, 2 patients had posterior base tumors and 1 had a lower polar tumor. Average blood loss was 350 ml, warm ischemia time was 28.46 minutes, and postoperative stay was 4.55 days. Of 30 specimens for histopathology, 23 (76%) were clear cell renal cell carcinoma (RCC), 4 (13%) were papillary RCC, 1 (3.3%) was chromophobe RCC, whereas 2 (6.6%) were benign (oncocytoma). Margins were free of tumors in all the patients with no recurrence in 2 years of follow-up. CONCLUSION: Laparoscopic partial nephrectomy by using a Satinsky clamp as a tool for en bloc hilar clamping in the proper axis at the hilum takes care of multiple vessels irrespective of size and number, particularly when renal angiography is not available. This technique of en bloc hilar clamping is quite useful especially in developing countries where robotic facilities are not available. The Satinsky clamp decreases blood loss and intraoperative time.

Comparison of percutaneous nephrolithotomy under epidural anesthesia versus general anesthesia: A randomized prospective study.

INTRODUCTION: PCNL has revolutionized the treatment of renal calculi putting almost an end to the era of open stone surgery. The procedure can safely be carried out under general anesthesia (GA) or regional anesthesia viz. spinal anesthesia (SA), epidural anesthesia (EA) or combined spinal and epidural anesthesia (CSE). AIMS AND OBJECTIVES: We evaluated the surgical outcome after PCNL in two groups of patients randomly divided to undergo procedure under GA or EA. PATIENTS AND METHODS: Two hundred and thirty patients with American Society of Anesthesiologists (ASA) score <3 were randomly divided into two groups according to the type of anesthesia: i.e. GA (n=110) or EA (n=120). All patients underwent PCNL in prone position. Puncture was done using Bulls eye technique under fluoroscopic guidance and tract dilated using serial dilators up to 24Fr-28 Fr. Demographics, perioperative and postoperative parameters were noted and data analysed. RESULTS: The two groups were comparable in terms of mean age, distribution of stone location, and stone burden. The stone free rate was 90.9% in GA group and 89.2% in EA group and the difference was statistically insignificant (P= 0.659). The requirement for auxiliary procedures was similar between the two groups. A significant difference in pain score was seen in favor of EA group during early post-operative period (P< 0.05). CONCLUSION: It seems that PCNL can be performed safely and effectively under regional epidural anesthesia with results comparable to general anesthesia with the added advantage of less immediate postoperative pain and analgesic requirement.

Surgical Outcome of Renal Cell Carcinoma with Tumor Thrombus Extension into Inferior Vena Cava and Right Atrium (Beating Heart Removal of Level 4 Thrombus): A Challenging Scenario.

AIM: "To evaluate oncological and surgical outcomes of different levels of tumor thrombus and tumor characteristics secondary to renal cell carcinoma (RCC)". MATERIALS AND METHODS: Retrospective review from 2013 to 2020 of 34 patients who underwent radical nephrectomy with thrombectomy for RCC with tumor thrombus extending into the inferior vena cava (IVC) and right atrium (RA) at our center. Level I and most level II tumors were removed using straight forward occluding maneuvers with control of the contralateral renal vein. None of the patients had level III tumor extensions in our study group. For level IV thrombus, a beating heart surgery using a simplified cardiopulmonary bypass (CPB) technique was used for retrieval of thrombus from the right atrium. RESULTS: " Of the 34 patients with thrombus", 19 patients had level I, 12 patients had level II, none had level III, and three patients had level IV thrombus. Two patients required simplified CPB. Another patient with level IV thrombus CPB, was not attempted in view of refractory hypotension intraoperatively. Pathological evaluation showed clear-cell carcinoma in 67.64%, papillary carcinoma in 17.64%, chromophobe in 5.8%, and squamous cell carcinoma in 8.8% of cases. Left side thrombectomy was difficult surgically, whereas right side thrombectomy did not have any survival advantage. Mean blood loss during the procedure was 325 mL, ranging from 200 to 1000 mL, and mean operative time was 185 min, ranging from 215 to 345 min. The immediate postoperative mortality was 2.9%. Level I thrombus had better survival compared to level II thrombus. CONCLUSION: Radical nephrectomy with tumor thrombectomy remains the mainstay of treatment in RCC with inferior venacaval extension. The surgical approach and outcome depends on primary tumor size, location, level of thrombus, local invasion of IVC, any hepato-renal dysfunction or any associated comorbidities. The higher the level of thrombus, the greater is the need for prior optimization and the adoption of a multidisciplinary approach for a successful surgical outcome.

A significant association of the CTLA4 gene variants with the risk of autoimmune Graves' disease in ethnic Kashmiri population.

Graves' disease (GD) is the commonest cause of hyperthyroidism in populations with adequate iodine intake. It results from an abnormality in the immune system, which produces unique antibodies causing over production of thyroid hormones and glandular hyperplasia in individuals with genetic susceptibility. The Cytotoxic Lymphocyte Associated Antigen-4 (CTLA4) gene product serves the important function of immunomodulation, thereby helping in maintenance of peripheral self-tolerance. Studies on the association of the CTLA4 SNPs with GD have shown variations in the results from different populations. Since no such study has been carried out in ethnic Kashmiri population, we aimed to study a possible association of the CTLA4 SNPs (+49 A/G, -318C/T, CT 60 A/G and -1661 A/G) with GD. A total of 285 individuals (135 patients with GD and 150 healthy individuals) were genotyped using PCR-RFLP method and the results showed statistically significant differences in genotypic and allelic frequencies of cases and controls for + 49 A/G SNP (p=<0.001; OR = 5.14; CI = 2.17-12.19) and CT 60 A/G SNP (p = < 0.001; OR = 6.9; CI = 2.8-16.6), while -318C/T and -1661 A/G SNPs showed no significant association. We also studied the mRNA expression of the CTLA4 in patients with GD and healthy individuals by Real-Time PCR and found a decreased expression of the CTLA4 mRNA in PBMCs of patients with GD as compared to healthy controls with a -3.71-fold change. We conclude that the CTLA4 + 49 A/G and CT 60 A/G SNPs have a significant association with the risk of GD development in Kashmiri population and CTLA4 mRNA expression is significantly decreased in GD.

Comparison between peak systolic velocity of the inferior thyroid artery and technetium-99m pertechnetate thyroid uptake in differentiating Graves' disease from thyroiditis

ABSTRACT Objectives The differentiation between the various etiologies of thyrotoxicosis, including those with hyperthyroidism (especially Graves' disease [GD], the most common cause of hyperthyroidism) and without hyperthyroidism (like thyroiditis), is an important step in planning specific therapy. Technetium-99m (99mTc) pertechnetate thyroid scanning is the gold standard in differentiating GD from thyroiditis. However, this technique has limited availability, is contraindicated in pregnancy and lactation, and is not helpful in cases with history of recent exposure to excess iodine. The aim of this study was to identify the diagnostic value of the peak systolic velocity of the inferior thyroid artery (PSV-ITA) assessed by color-flow Doppler ultrasound (CFDU) and compare the sensitivity and specificity of this method versus 99mTc pertechnetate thyroid uptake. Subjects and methods We prospectively analyzed 65 patients (46 with GD and 19 with thyroiditis). All patients were evaluated with clinical history and physical examination and underwent 99mTc pertechnetate scanning and measurement of TRAb levels and PSV-ITA values by CFDU. The diagnosis was based on findings from signs and symptoms, physical examination, and 99mTc pertechnetate uptake. Results Patients with GD had significantly higher mean PSV-ITA values than those with thyroiditis. At a mean PSV-ITA cutoff value of 30 cm/sec, PSV-ITA discriminated GD from thyroiditis with a sensitivity of 91% and specificity of 89%. Conclusion Measurement of PSV-ITA by CFDU is a good diagnostic approach to discriminate between GD and thyroiditis, with sensitivity and specificity values comparable to those of 99mTc pertechnetate thyroid uptake.

Comparison between peak systolic velocity of the inferior thyroid artery and technetium-99m pertechnetate thyroid uptake in differentiating Graves' disease from thyroiditis.

OBJECTIVES: The differentiation between the various etiologies of thyrotoxicosis, including those with hyperthyroidism (especially Graves' disease [GD], the most common cause of hyperthyroidism) and without hyperthyroidism (like thyroiditis), is an important step in planning specific therapy. Technetium-99m (99mTc) pertechnetate thyroid scanning is the gold standard in differentiating GD from thyroiditis. However, this technique has limited availability, is contraindicated in pregnancy and lactation, and is not helpful in cases with history of recent exposure to excess iodine. The aim of this study was to identify the diagnostic value of the peak systolic velocity of the inferior thyroid artery (PSV-ITA) assessed by color-flow Doppler ultrasound (CFDU) and compare the sensitivity and specificity of this method versus 99mTc pertechnetate thyroid uptake. SUBJECTS AND METHODS: We prospectively analyzed 65 patients (46 with GD and 19 with thyroiditis). All patients were evaluated with clinical history and physical examination and underwent 99mTc pertechnetate scanning and measurement of TRAb levels and PSV-ITA values by CFDU. The diagnosis was based on findings from signs and symptoms, physical examination, and 99mTc pertechnetate uptake. RESULTS: Patients with GD had significantly higher mean PSV-ITA values than those with thyroiditis. At a mean PSV-ITA cutoff value of 30 cm/sec, PSV-ITA discriminated GD from thyroiditis with a sensitivity of 91% and specificity of 89%. CONCLUSION: Measurement of PSV-ITA by CFDU is a good diagnostic approach to discriminate between GD and thyroiditis, with sensitivity and specificity values comparable to those of 99mTc pertechnetate thyroid uptake.

Vitexin alleviates non-alcoholic fatty liver disease by activating AMPK in high fat diet fed mice.

Non-alcoholic fatty liver disease (NAFLD) is a most common liver disorder characterized by accumulation of fat in the liver and currently there is no approved treatment for it. Obesity and diabetes being leading cause of NAFLD, compounds having anti-obesity activity and potential to reduce insulin resistance are considered suitable candidate for NAFLD treatment. In this study, we checked effect of vitexin, a naturally occurring flavonoid, on high fat diet (HFD) induced NAFLD in C57BL/6J mice. In presence of vitexin, significant reduction in body and liver weight, triglyceride and cholesterol content in serum and liver was observed. Serum Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) levels were reduced significantly by vitexin which were elevated in HFD group whereas serum lipase activity remained unchanged. Vitexin suppressed de novo lipogenesis by downregulating expression of Peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein-α (C/EBP-α), sterol regulatory element-binding protein-1c (SREBP-1c), Fatty acid synthase (FAS) and Acetyl-CoA Carboxylase (ACC). Additionally, it also enhanced fatty acid oxidation and lipolysis by upregulating Peroxisome proliferator-activated receptor α (PPAR-α), carnitine palmitoyltransferase-1a (CPT-1a) and Adipose triglyceride lipase (ATGL). Inhibition of lipogenesis and activation of lipolysis and fatty acid oxidation by vitexin was found to be mediated by activation of AMP-activated protein kinase (AMPK). Vitexin also improved insulin signalling by activating insulin receptor substrate-1 (IRS-1) and its downstream target AKT. AMPK activation of vitexin was possibly through binding of vitexin to leptin receptor (LepR) which was confirmed by molecular docking studies and by observed enhanced expression of LepR. Thus, we propose that vitexin alleviates NAFLD by activating AMPK possibly by binding to LepR.

Pterostilbene reverses palmitic acid mediated insulin resistance in HepG2 cells by reducing oxidative stress and triglyceride accumulation.

Insulin resistance (IR) is known to precede onset of type 2 diabetes and increased oxidative stress appears to be a deleterious factor leading to IR. In this study, we evaluated ability of pterostilbene (PTS), a methoxylated analogue of resveratrol and a known antioxidant, to reverse palmitic acid (PA)-mediated IR in HepG2 cells. PTS prevented reactive oxygen species (ROS) formation and subsequent oxidative lipid damage by reducing the expression of NADPH oxidase 3 (NOX3) in PA treated HepG2 cells. Hepatic glucose production was used as a measure of IR and PTS reversed PA-mediated increase in hepatic glucose production by reducing expression of genes coding for gluconeogenic enzymes namely glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate carboxylase (PC); and their transcription factors cAMP response element binding protein (CREB) and fork head class Box O (FOXO1) along with its coactivator peroxisome proliferator-activated receptor gamma co-activator-1 α (PGC1α). PTS reversed PA-mediated activation of c-Jun N-terminal kinase (JNK), which in turn altered insulin signalling pathway by phosphorylating IRS-1 at Ser 307, leading to inhibition of phosphorylation of Akt and GSK-3ß. PTS also reduced PA-mediated lipid accumulation by reducing expression of transcription factors SREBP1c and PPARα. SREBP1c activates genes involved in fatty acid and triglyceride synthesis while PPARα activates CPT1, a rate limiting enzyme for controlling entry and oxidation of fatty acids into mitochondria. PTS, however, did not influence PA uptake confirmed by using BODIPY-labelled fluorescent C16 fatty acid analogue. Thus, our data provides a possible mechanistic explanation for reversal of PA-mediated IR in HepG2 cells.

PTPN22 1858 C/T Exon Polymorphism is not Associated with Graves' Disease in Kashmiri population.

BACKGROUND: Graves' disease (GD) is a multifactorial autoimmune disease with contribution from both genetic and epigenetic factors in its causation. Association of genetic factors and GD has been extensively studied. Gene "protein tyrosine phosphatase nonreceptor 22" (PTPN22) is an important immunoregulatory gene preventing hyper responsiveness of T cells by negatively regulating their signal transduction. Association of single-nucleotide polymorphism (SNP) 1858 C/T within PTPN22 with some autoimmune diseases has been described. METHODS: We aimed to analyze whether 1858 C/T SNP of PTPN22 gene has any association with GD in Kashmiri population. Polymerase chain reaction-restriction fragment length polymorphism was performed for genotyping 1858 C/T SNP in 135 patients with GD and 150 age- and gender-matched healthy controls. RESULTS: Among the patients with GD, the frequencies of PTPN22 1858 CC, CT, and TT genotypes were 97.7, 2.2, and 0%, respectively, whereas in healthy controls the frequencies of CC, CT genotypes were 100 and 0%, respectively. No significant association was found between PTPN22 1858 C/T SNP and patients with GD. CONCLUSION: GD is not associated with PTPN22 1858 C/T SNP in Kashmiri population. Furthermore, 1858 C/T SNP in PTPN22 gene could be a part of variation in different ethnic populations across the globe.

Association of FoxP3 promoter polymorphisms with the risk of Graves' disease in ethnic Kashmiri population.

Graves' disease is a multifactorial autoimmune disorder of the thyroid gland, with some extra-thyroidal complications like eye and skin abnormalities in some patients. GD is more prevalent in women than men and is the leading cause of hyperthyroidism worldwide. A complex interaction between genetic and environmental factors is the proposed cause which triggers immune system to produce autoantibodies stimulating the TSH receptor, leading to clinical manifestations such as hyperthyroidism, diffuse thyroid enlargement (goiter) and often ophthalmopathy in affected individuals. Various Single nucleotide gene polymorphisms (SNPs) have been associated with the risk of GD development including promoter SNPs in Forkhead Box P3 (FOXP3). FOXP3 is an important regulatory factor for T cell development and differentiation and therefore has a prominent role in suppression of autoimmune reactions which may lead to predisposition of GD. There have been some studies on the association of FOXP3 SNPs with GD, but no such investigation has been carried out in ethnic Kashmiri population. So, we aimed to study a possible association of FOXP3 promoter SNPs (-3279C/A, -2383C/T & -3499 A/G) with GD. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) was used to genotype 285 individuals (135 GD cases and 150 healthy controls) and the results showed statistically significant differences in genotypic and allelic frequencies of cases and controls for -3279C/A SNP [OR, 3.48; 95% CI (2.05-5.92); P < 0.001] and -2383C/T SNP [OR, 5.62; 95% CI (2.43-13.00); P < 0.001], while no significant association was seen in case of -3499 A/G SNP. We conclude that -3279C/A and -2383C/T SNPs have a highly significant association with the risk of GD development in Kashmiri population.

Association of polymorphic variants of IL-1ß and IL-1RN genes in the development of Graves' disease in Kashmiri population (North India).

PURPOSE: Graves' disease (GD) is a multigenic, organ specific autoimmune disorder with a strong genetic predisposition and IL-1ß has been shown to be involved in its pathogenesis. The present study was aimed to determine the genetic associations between polymorphisms of IL-1ß gene promoter region (-511 T>C) (rs16944), exon 5 (+3954 C>T) (rs1143634) and IL-1RN gene VNTR (rs2234663) polymorphism in patients with GD in ethnic Kashmiri population. METHODS: A total of 135 Graves' disease patients and 150 healthy individuals were included in the study. PCR and PCR-based restriction analysis methods were done for IL-1RNVNTR and IL-1ß gene polymorphisms respectively. RESULTS: We found statistically significant increased frequencies of the C/C + CT genotype (P = 0.001; odds ratio (OR) = 5.04, 95% confidence interval (CI) = 3.02-8.42) and the C allele (P = 0.001; OR = 3.10, 95% CI = 2.14-4.50) in IL-1ß gene promoter polymorphism (rs16944) with GD patients compared to normal controls. Also in the exon 5 (rs1143634), a significant increase in frequency of the C/C homozygous genotype (P = 0.001; OR = 0.18, 95% CI = 0.11-0.30) and C allele (P = 0.001; OR = 0.31, 95% CI = 0.20-0.48) was observed in GD cases as against controls. For IL-1RNVNTR (rs2234663), we didn't observe any significant difference in the allelic and genotypic frequencies between cases and controls. CONCLUSION: Our findings suggest that both promoter and exon polymorphisms of IL-1ß gene have a significant role in the risk of developing GD, whereas IL-1RNVNTR has no association with GD.