Fluoride export is required for the competitive fitness of pathogenic microorganisms in dental biofilm models.

Microorganisms resist fluoride toxicity using fluoride export proteins from one of several different molecular families. Cariogenic species Streptococcus mutans and Candida albicans extrude intracellular fluoride using a CLCF F-/H+ antiporter and FEX fluoride channel, respectively, whereas oral commensal eubacteria, such as Streptococcus gordonii, export fluoride using a Fluc fluoride channel. In this work, we examine how genetic knockout of fluoride export impacts pathogen fitness in single-species and three-species dental biofilm models. For biofilms generated using S. mutans with the genetic knockout of the CLCF transporter, exposure to low fluoride concentrations decreased S. mutans counts, synergistically reduced the populations of C. albicans, increased the relative proportion of oral commensal S. gordonii, and reduced properties associated with biofilm pathogenicity, including acid production and hydroxyapatite dissolution. Biofilms prepared with C. albicans with genetic knockout of the FEX channel also exhibited reduced fitness in the presence of fluoride but to a lesser degree. Imaging studies indicate that S. mutans is highly sensitive to fluoride, with the knockout strain undergoing complete lysis when exposed to low fluoride for a moderate amount of time. Biochemical purification of the S. mutans CLCF transporter and functional reconstitution establishes that the functional protein is a dimer encoded by a single gene. Together, these findings suggest that fluoride export by oral pathogens can be targeted by specific inhibitors to restore biofilm symbiosis in dental biofilms and that S. mutans is especially susceptible to fluoride toxicity. IMPORTANCE: Dental caries is a globally prevalent condition that occurs when pathogenic species, including Streptococcus mutans and Candida albicans, outcompete beneficial species, such as Streptococcus gordonii, in the dental biofilm. Fluoride is routinely used in oral hygiene to prevent dental caries. Fluoride also has antimicrobial properties, although most microbes possess fluoride exporters to resist its toxicity. This work shows that sensitization of cariogenic species S. mutans and C. albicans to fluoride by genetic knockout of fluoride exporters alters the microbial composition and pathogenic properties of dental biofilms. These results suggest that the development of drugs that inhibit fluoride exporters could potentiate the anticaries effect of fluoride in over-the-counter products like toothpaste and mouth rinses. This is a novel strategy to treat dental caries.

The yeast AMP-activated protein kinase Snf1 phosphorylates the inositol polyphosphate kinase Kcs1.

The yeast Snf1/AMP-activated kinase (AMPK) maintains energy homeostasis, controlling metabolic processes and glucose derepression in response to nutrient levels and environmental cues. Under conditions of nitrogen or glucose limitation, Snf1 regulates pseudohyphal growth, a morphological transition characterized by the formation of extended multicellular filaments. During pseudohyphal growth, Snf1 is required for wild-type levels of inositol polyphosphate (InsP), soluble phosphorylated species of the six-carbon cyclitol inositol that function as conserved metabolic second messengers. InsP levels are established through the activity of a family of inositol kinases, including the yeast inositol polyphosphate kinase Kcs1, which principally generates pyrophosphorylated InsP7. Here, we report that Snf1 regulates Kcs1, affecting Kcs1 phosphorylation and inositol kinase activity. A snf1 kinase-defective mutant exhibits decreased Kcs1 phosphorylation, and Kcs1 is phosphorylated in vivo at Ser residues 537 and 646 during pseudohyphal growth. By in vitro analysis, Snf1 directly phosphorylates Kcs1, predominantly at amino acids 537 and 646. A yeast strain carrying kcs1 encoding Ser-to-Ala point mutations at these residues (kcs1-S537A,S646A) shows elevated levels of pyrophosphorylated InsP7, comparable to InsP7 levels observed upon deletion of SNF1. The kcs1-S537A,S646A mutant exhibits decreased pseudohyphal growth, invasive growth, and cell elongation. Transcriptional profiling indicates extensive perturbation of metabolic pathways in kcs1-S537A,S646A. Growth of kcs1-S537A,S646A is affected on medium containing sucrose and antimycin A, consistent with decreased Snf1p signaling. This work identifies Snf1 phosphorylation of Kcs1, collectively highlighting the interconnectedness of AMPK activity and InsP signaling in coordinating nutrient availability, energy homoeostasis, and cell growth.

Fluoride export is required for competitive fitness of pathogenic microorganisms in dental biofilm models.

Microorganisms resist fluoride toxicity using fluoride export proteins from one of several different molecular families. Cariogenic species Streptococcus mutans and Candida albicans extrude intracellular fluoride using a CLCF F-/H+ antiporter and FEX fluoride channel, respectively, whereas commensal eubacteria, such as Streptococcus gordonii, export fluoride using a Fluc fluoride channel. In this work, we examine how genetic knockout of fluoride export impacts pathogen fitness in single-species and three-species dental biofilm models. For biofilms generated using S. mutans with genetic knockout of the CLCF transporter, exposure to low fluoride concentrations decreased S. mutans counts, synergistically reduced the populations of C. albicans, increased the relative proportion of commensal S. gordonii, and reduced properties associated with biofilm pathogenicity, including acid production and hydroxyapatite dissolution. Biofilms prepared with C. albicans with genetic knockout of the FEX channel also exhibited reduced fitness in the presence of fluoride, but to a lesser degree. Imaging studies indicate that S. mutans is highly sensitive to fluoride, with the knockout strain undergoing complete lysis when exposed to low fluoride for a moderate amount of time, and biochemical purification the S. mutans CLCF transporter and functional reconstitution establishes that the functional protein is a dimer encoded by a single gene. Together, these findings suggest that fluoride export by oral pathogens can be targeted by specific inhibitors to restore biofilm symbiosis in dental biofilms, and that S. mutans is especially susceptible to fluoride toxicity.

Streeter's Syndrome of Lower Limb Associated with CTEV.

Introduction: The term Streeter's syndrome is a term used to describe rare congenital malformations that includes a variety of clinical presentations usually consisting of a constriction band around a part of the body which can be as superficial as involving just the skin which can be only cosmetic and asymptomatic or can be as deep as causing restricted circulation distally which may be in incompatible with life. Such conditions are remarkably rare accounting for an incidence range from 1:1.2 k to 1: 15 k live births and 178:10 k spontaneous abortions [1]. Males and females are uniformly affected. Almost all cases are sporadic; extremely rare evidence of familial transmission. The entity has been described in the literature in 34 different terms, (such as amniotic rupture sequence, ADAM complex, constriction band syndrome, Streeter's dysplasia, etc.) due to its extremely variable clinical features and lack of understanding of the etiology. This results in a lack of understanding and creates unnecessary stress for the surgeon/physician as well as the parents. Case Report: We discuss case reports of two such cases with their simple nature of treatment with their outcomes. Conclusion: There is a significant deficit in the education of cases with low incidence, such is the case with pediatric patients presenting with amniotic bands, which usually present and are associated with Congenital Talipes Equino Varus deformity. In such cases, improper or incorrect treatment and/or neglect of the constriction may lead to the vascular deficit and eventually auto-amputation of the segment distal to the amniotic band.

Prevalence of Vascular Calcification in Chronic Kidney Disease Stage 4 and 5 Patients and its Correlation with Inflammatory Markers of Atherosclerosis.

Vascular calcification is one of the independent risk factors associated with cardiovascular disease (CVD) in chronic kidney disease (CKD) patients. This study evaluated the prevalence of vascular calcification in Indian patients with CKD stages 4 and 5. This was a prospective study conducted between January 2011 and June 2012. CKD stage 4 and 5 patients of either sex aged >18 years were screened for aortic vascular calcification using digital X-ray lumbar spine and multislice computed tomography (CT) scan. In addition, details of inflammatory markers [high-sensitivity C-reactive protein (hs-CRP) and interleukin (IL-6)] were also collected. A total of 150 patients (stage 4, n = 98; stage 5, n = 56) were screened for vascular calcification, and the mean age was 56.56 years. Patients with CKD stage 5 had significantly higher (P ≤0.05) serum creatinine and lower estimated glomerular filtration rate, total cholesterol, and low-density lipoprotein than CKD stage 4. Significantly, more patients with CKD stage 5 had a history of CVD. A total of 113 (75.3%) patients had vascular calcification [aortic calcification index (ACI) >0] with significantly higher prevalence in CKD stage 5 (85.72%) as compared to CKD stage 4 (69.15%). Patients having high aortic calcification (ACI >20%) were older (P = 0.0013); had a higher frequency of diabetes, and CVD; and had significantly (P <0.05) higher blood urea, serum creatinine, phosphorus, Ca × PO4 product, intact parathyroid hormone, hs-CRP, and IL-6. The higher CKD stage was associated with a higher prevalence of vascular calcification and higher aortic calcification index (ACI). CT techniques (electron beam CT and multislice CT) are the gold standards for detection and quantification of progression of vascular calcification.

Double-strand breaks in motion: implications for chromosomal rearrangement.

DNA double-strand breaks (DSBs) must be rejoined properly to prevent the occurrence of serious genomic rearrangements associated with many human diseases. Non-homologous end joining (NHEJ) is a DSB repair mechanism known to protect genomic integrity that is also implicated in creating genomic translocations, inversions, deletions, and insertions. We recently investigated the impact of the pre-damage spatial proximity of DSB-bearing loci on the frequency of trans repair by NHEJ and surprisingly found no correlation between them. In this review, we consider various models that might account for these unexpected results. While DSB movement is necessary to explain our findings, many questions remain about the nature and timing of that motion.

Frequency of DNA end joining in trans is not determined by the predamage spatial proximity of double-strand breaks in yeast.

DNA double-strand breaks (DSBs) are serious genomic insults that can lead to chromosomal rearrangements if repaired incorrectly. To gain insight into the nuclear mechanisms contributing to these rearrangements, we developed an assay in yeast to measure cis (same site) vs. trans (different site) repair for the majority process of precise nonhomologous end joining (NHEJ). In the assay, the HO endonuclease gene is placed between two HO cut sites such that HO expression is self-terminated upon induction. We further placed an additional cut site in various genomic loci such that NHEJ in trans led to expression of a LEU2 reporter gene. Consistent with prior reports, cis NHEJ was more efficient than trans NHEJ. However, unlike homologous recombination, where spatial distance between a single DSB and donor locus was previously shown to correlate with repair efficiency, trans NHEJ frequency remained essentially constant regardless of the position of the two DSB loci, even when they were on the same chromosome or when two trans repair events were put in competition. Repair of similar DSBs via single-strand annealing of short terminal direct repeats showed substantially higher repair efficiency and trans repair frequency, but still without a strong correlation of trans repair to genomic position. Our results support a model in which yeast cells mobilize, and perhaps compartmentalize, multiple DSBs in a manner that no longer reflects the predamage position of two broken loci.

Overhang polarity of chromosomal double-strand breaks impacts kinetics and fidelity of yeast non-homologous end joining.

Non-homologous end joining (NHEJ) is the main repair pathway for DNA double-strand breaks (DSBs) in cells with limited 5' resection. To better understand how overhang polarity of chromosomal DSBs affects NHEJ, we made site-specific 5'-overhanging DSBs (5' DSBs) in yeast using an optimized zinc finger nuclease at an efficiency that approached HO-induced 3' DSB formation. When controlled for the extent of DSB formation, repair monitoring suggested that chromosomal 5' DSBs were rejoined more efficiently than 3' DSBs, consistent with a robust recruitment of NHEJ proteins to 5' DSBs. Ligation-mediated qPCR revealed that Mre11-Rad50-Xrs2 rapidly modified 5' DSBs and facilitated protection of 3' DSBs, likely through recognition of overhang polarity by the Mre11 nuclease. Next-generation sequencing revealed that NHEJ at 5' DSBs had a higher mutation frequency, and validated the differential requirement of Pol4 polymerase at 3' and 5' DSBs. The end processing enzyme Tdp1 did not impact joining fidelity at chromosomal 5' DSBs as in previous plasmid studies, although Tdp1 was recruited to only 5' DSBs in a Ku-independent manner. These results suggest distinct DSB handling based on overhang polarity that impacts NHEJ kinetics and fidelity through differential recruitment and action of DSB modifying enzymes.

A prospective, multi-centre, observational study to examine kidney disease progression in adults with chronic kidney disease - CKDOD - Study design and preliminary results.

BACKGROUND: The objective of this article is to describe the organisation of an international, clinical registry, the Chronic Kidney Disease Observational Database (CKDOD), the processes of enrolling patients and entering data and preliminary results to date. DESIGN: The Chronic Kidney Disease Observational Database (CKDOD) is designed to assess the association between different factors with a known influence on chronic kidney disease (CKD) progression as well as treatment strategies such as dietary modifications, blood pressure control and pharmacological interventions in Asian countries (India, China, Malaysia and Thailand). The only inclusion criterion is the presence of CKD stage 2 or higher as defined by the KDIGO guidelines. Demographic and clinical information are collected by a standardised electronic questionnaire, available in English and Chinese. The data are transferred to the CKDOD database either by e-mail or via web access. All data are checked for consistency and missing values. Collection of data started in September 2011 and by April 2015, data on 1323 individual patients had been submitted. The mean age at inclusion was 57 ± 14 years, 67 % were male and 36 % were diabetic. The baseline estimated glomerular filtration rate was 26 ml/min/1.73 m(2). Of all enrolled patients, 324 (24 %) received ketoanalogue supplementation during at least one recorded visit. DISCUSSION: The CKDOD is a very large and comprehensive data repository, currently focused in subjects recruited from Asia. The database is expected to provide important long-term information on CKD progression, nutritional and metabolic derangements that accompany CKD progression and treatment strategies to ameliorate progression and complications of CKD. TRIAL REGISTRATION: Clinical Trial Registry - India: CTRI/2012/06/002743 ; 25th July 2012.

Unusual metastatic presentation of carcinoma hypopharynx.

Cutaneous metastases from hypopharyngeal cancers is rare constituting about 0.8-1.3% and represent a sub-group of head and neck cancer patients who have very poor prognosis even when treated. We report a case of 65-year-old male diagnosed as carcinoma hypo pharynx stage IV who was on radiotherapy when he developed cutaneous metastasis over the chest wall, which initially presented as small nodules and later progressed into a proliferative lesion. Patient received further radiation to the metastatic lesion, but the disease was progressive, demonstrating that head and neck squamous cell cancer patients with skin metastasis fare poorly.

A rare case of type-I auto-immune hepatitis and thyroiditis presenting with crescentic glomerulonephritis.

A middle-aged female patient with a past history of non-alcoholic liver disease and hypothyroidism presented with swelling of the body, off and on, for six months and rapidly worsening renal function. Renal biopsy showed crescentic glomerulonephritis with negative immunofluorescence. Serological tests were positive for anti-thyroglobulin, anti-nuclear antibody (1:80), p-anti-neutrophil cytoplasmic antibodies; gamma globulin was 5.23 g/dL and viral markers were negative. The patient was diagnosed to have autoimmune hepatitis type-1 and treated with injection methylprednisolone pulse (500 mg/day for 3 days) and maintained on oral steroids and azathioprine 100 mg. She responded dramatically to this treatment and has remained in complete remission at last follow-up.

Autophosphorylation of the Smk1 MAPK is spatially and temporally regulated by Ssp2 during meiotic development in yeast.

Smk1 is a meiosis-specific MAPK that controls spore wall morphogenesis in Saccharomyces cerevisiae. Although Smk1 is activated by phosphorylation of the threonine (T) and tyrosine (Y) in its activation loop, it is not phosphorylated by a dual-specificity MAPK kinase. Instead, the T is phosphorylated by the cyclin-dependent kinase (CDK)-activating kinase, Cak1. The Y is autophosphorylated in an intramolecular reaction that requires a meiosis-specific protein named Ssp2. The meiosis-specific CDK-like kinase, Ime2, was previously shown to positively regulate Smk1. Here we show that Ime2 activity is required to induce the translation of SSP2 mRNA at anaphase II. Ssp2 protein is then localized to the prospore membrane, the structure where spore wall assembly takes place. Next the carboxy-terminal portion of Ssp2 forms a complex with Smk1 and stimulates the autophosphorylation of its activation-loop Y residue. These findings link Ime2 to Smk1 activation through Ssp2 and define a developmentally regulated mechanism for activating MAPK at specific locations in the cell.

Evaluation of Protein-Energy Wasting and Inflammation on Patients Undergoing Continuous Ambulatory Peritoneal Dialysis and its Correlations.

BACKGROUND: Protein-energy wasting (PEW) and heightened inflammation are prevalent in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and is a strong risk factor for morbidity and mortality in these patients. Evaluation of PEW, prevalence of inflammation as well as interrelationship between various nutritional indices and inflammation has not been studied in much detail in patients undergoing CAPD. OBJECTIVES: This study was conducted to evaluate the interrelationship between PEW and inflammation in patients undergoing CAPD. PATIENTS AND METHODS: Sixty-three patients undergoing CAPD (M = 28, F = 35) were assessed with regard to their nutritional status and inflammation after a minimum of 3 months CAPD initiation. Nutritional status was assessed by dietary diary, anthropometry, subjective global assessment, and multifrequency bioelectrical impedance analysis (BIA). In addition, their serum albumin, prealbumin, transferrin, and cholesterol level were measured. Also, inflammation in these patients was assessed by High-Sensitivity C-Reactive Protein (hs-CRP > 3 mg/L) and Interleukin-6 (IL-6 > 2 µg/mL). Later on, diagnosis of malnutrition was made based on different methods. Correlation between inflammation and various nutritional assessment indices were analyzed statistically. RESULTS: Mean (SD) age of the patients was 57.6 (11.6) years. The average (SD) calorie and protein intake per day were 25.5 (4.6) kcal and 0.81 (0.2) mg, respectively. The mean and standard deviation of anthropometry variables of body mass index (BMI), mid-arm circumference (MAC), tricipital skin-fold thickness (TST), mid-arm muscle circumference (MAMC), and corrected mid-arm muscle area (cMAMA) were 23.7 ± 5 kg/m(2), 26.3 ± 4.5 cm, 1.624 ± 0.4 cm, 25.6 ± 4.5 cm, and 45.7 ± 19.7 cm(2), respectively. The mean values of serum protein, albumin, prealbumin, transferrin, cholesterol, triglyceride, hs-CRP, and IL-6 were 5.9 g/dL, 3.0 g/dL, 21.11 mg/dL, 130.6 mg/dL, 155.9 mg/dL, 136.1 mg/dL, 8.8 ± 7.6 mg/L, and 8.4 ± 12.2 µg/dL, respectively. Based on subjective global assessment (SGA); 11.63 (17.4%), 34.63 (54%), and 18.65 (28.6%) patients undergoing CAPD had normal, moderate, and severe malnutrition status, respectively. According to serum albumin level; 13.63 (21%), 39.63 (62%), and 11.63 (17%) patients undergoing CAPD had normal, moderate, and severe malnutrition status, respectively. Finally, based on BMI; 33.63 (52%), 23.63 (37%), and 7.63 (11%) patients undergoing CAPD had normal, moderate, and severe malnutrition status, respectively. About 76.1% and 9.5% of patients undergoing CAPD were malnourished based on lean tissue index (LTI) and fat tissue index (FTI), respectively. Based on hs-CRP and IL-6 findings, 70% (44/63) and 71.8% (45/63) of patients undergoing CAPD had high inflammation, respectively. High sensitive C-reactive protein correlated negatively (significantly) with serum albumin, prealbumin, and transferrin. Interleukin -6 correlated negatively (significantly) with MAC; MAMA; serum albumin, cholesterol, and transferrin. There was significant positive correlation between hs-CRP and IL-6. There is statistically significant difference in total protein intake (g/d), protein intake (g/kg/d), serum protein (g/dL), albumin (g/dL), transferrin (mg/dL), and cholesterol (mg/dL) between patients with and without inflammation. CONCLUSIONS: Protein-energy wasting (80% - 85%) by various methods and inflammation (70%) was very prevalent among patients undergoing CAPD. Inflammatory markers show significant negative correlation with anthropometry and serological markers. Inflammatory markers are suggested to be included in the regular assessment of patients undergoing CAPD, for the better management of protein-energy wasting.

Estimation of renal function in the intensive care unit: the covert concepts brought to light.

Frantic efforts have been made up to this date to derive consensus for estimating renal function in critically ill patients, only to open the Pandora's box. This article tries to explore the various methods available to date, the newer concepts, and the uncared issues that may still prove to be useful in estimating renal function in intensive care unit patients. The concept of augmented renal clearance, which is frequently encountered in critically ill patients, should always be taken into account, as correct therapeutic dosage of drugs sounds vital which in turn depends on correctly calculated glomerular filtration rate. Serum creatinine and creatinine-based formulae have their own demerits that are well known and established. While Cockcroft-Gault and 4-variable modification of diet in renal diseases formulae are highly inadequate in the intensive care setup for estimating glomerular filtration rate, employing isotopic methods is impractical and cumbersome. The 6-variable modification of diet in renal diseases formula fairs better as it takes into account the serum albumin and blood urea nitrogen, too. Jelliffe's and modified Jelliffe's equations take into account the rate of creatinine production and volume of distribution which in turn fluctuates heavily in a critically ill patient. Twenty-four-hour and timed creatinine clearances offer values close to reality although not accurate and cannot provide immediate results. Cystatin C is a novel agent that offers a sure promise as it is least influenced by factors that affect serum creatinine to a major extent. Aminoglycoside clearance, although still in the dark area, may prove a simple yet precise way of estimating glomerular filtration rate in those patients in whom these drugs are therapeutically employed. Optic ratiometric method has emerged as the most sophisticated one in glomerular filtration rate estimation in critically ill patients.

Yeast DNA ligase IV mutations reveal a nonhomologous end joining function of BRCT1 distinct from XRCC4/Lif1 binding.

LIG4/Dnl4 is the DNA ligase that (re)joins DNA double-strand breaks (DSBs) via nonhomologous end joining (NHEJ), an activity supported by binding of its tandem BRCT domains to the ligase accessory protein XRCC4/Lif1. We screened a panel of 88 distinct ligase mutants to explore the structure­function relationships of the yeast Dnl4 BRCT domains and inter-BRCT linker in NHEJ. Screen results suggested two distinct classes of BRCT mutations with differential effects on Lif1 interaction as compared to NHEJ completion. Validated constructs confirmed that D800K and GG(868:869)AA mutations, which target the Lif1 binding interface, showed a severely defective Dnl4­Lif1 interaction but a less consistent and often small decrease in NHEJ activity in some assays, as well as nearly normal levels of Dnl4 accumulation at DSBs. In contrast, mutants K742A and KTT(742:744)ATA, which target the ß3-α2 region of the first BRCT domain, substantially decreased NHEJ function commensurate with a large defect in Dnl4 recruitment to DSBs, despite a comparatively greater preservation of the Lif1 interaction. Together, these separation-of-function mutants indicate that Dnl4 BRCT1 supports DSB recruitment and NHEJ in a manner distinct from Lif1 binding and reveal a complexity of Dnl4 BRCT domain functions in support of stable DSB association.

Analysis of case series of milky urine: A single center and departmental clinical experience with emphasis on management perspectives: A prospective observational study.

BACKGROUND: Milky urine can be due to chyluria or lipiduria due to nephrotic syndrome. Filarial chyluria usually responds to medical management while non-filarial cases may require surgical intervention. AIM: To perform a prospective observational study in patients presenting with milky urine in our centre over a period of one year from July 2011 to June 2012, a complete biochemical work up and imaging to find out the site of leakage of lymph if it is a case of chyluria, its response to medical management and the requirement of surgical intervention. MATERIALS AND METHODS: Routine blood and urine investigations, 24 hour urine protein excretion, USG abdomen, serum lipid profile and rapid filarial antigen test were done in all. MRI abdomen was done in affordable patients. Renal biopsy was done in some chyluria patients for academic purpose and in milky urine with negative urine ether test. Sclerotherapy was done with 50% dextrose and 0.2% povidone iodine. Patients were followed up with 24 hour urine protein and triglyceride estimation. RESULTS: 18 cases of milky urine were encountered. 8 were filarial chyluria, 9 non- filarial and 1 MCD. Mean urine TG level and median 24 hour urinary protein excretion were 37.2 ± 24.6 mg% and 4.96 g respectively. The mean age for filariasis (22.9 ± 4.5 years) was significantly different from that of non-filarial etiology (31.5 ± 4.8 years) (P = 0.005). The mean 24 hour urinary protein for normal MRI cases (4.64 ± 0.70 g) was significantly different from those with dilated lymphatics (8.15 ± 2.55 g) (P = 0.02). All the non- filarial and 4 filarial cases required sclerotherapy. One patient required a second sitting. CONCLUSION: Milky urine is most commonly due to chyluria and occasionally due to nephrotic syndrome. Nephrotic syndrome is managed in its own way while chyluria not amenable to pharmacological intervention is managed with sclerotherapy.

Post cardiac surgery acute kidney injury: a woebegone status rejuvenated by the novel biomarkers.

BACKGROUND: Acute kidney injury (AKI) is common after cardiac surgery, the incidence varying between 7.7% and 28.1%. It significantly increases morbidity and mortality. Creatinine considerably delays the diagnosis with its own attended demerits. Novel urinary biomarkers are emerging which help in rapid diagnosis thus reducing the morbidity and mortality. Biomarkers of our study were neutrophil gelatinase-associated lipocalin (NGAL) and Interleukin-18 (IL-18). OBJECTIVES: To find out the incidence of AKI in post-cardiac surgery patients in our hospital, the ability of the two biomarkers in early diagnosis in predicting the severity of AKI based on RIFLE's criteria and their ability to discriminate pre-renal from intrinsic AKI. PATIENTS AND METHODS: One-hundred patients who underwent cardiac surgery were selected. Midstream urine samples were collected at 3 time intervals (baseline before surgery, 24 hours and 7 days after surgery). Biomarkers were measured by ELISA using BIORAD processors. Fractional excretion of sodium and urea were used to discriminate pre-renal from intrinsic AKI. RESULTS: Out of 100 patients, 31 had AKI, 11 being pre-renal and 20 intrinsic AKI. Four patients required renal replacement therapy (12.9% among AKI cases and 4% in the overall study cohort). Four among 31 expired in intensive care unit. Identifiable risk factors for AKI included insulin requiring diabetes mellitus, chronic obstructive pulmonary disease, increased cardio-pulmonary bypass time, combined valvular surgery and coronary artery bypass grafting, employment of intra-aortic balloon counter pulsation, left main coronary artery occlusion and an ejection fraction of < 40%. NGAL was extremely sensitive (area under curve-0.96) in detecting intrinsic AKI at 24 hours followed by IL-18 ratio with an area under curve of 0.89. Creatinine at 24 hours was able to detect only 31.6% of intrinsic AKI. None of the pre-renal cases showed rise in the urinary biomarker levels. Patients with higher stages of AKI had higher levels of both biomarkers than those at lower stages. CONCLUSIONS: NGAL and IL-18 obviated the disadvantages of creatinine. They were efficient in early detection of AKI, in differentiating pre-renal from intrinsic AKI and in predicting the severity of AKI reliably in post-cardiac surgery patients.

Multisite phosphorylation of the Sum1 transcriptional repressor by S-phase kinases controls exit from meiotic prophase in yeast.

Activation of the meiotic transcription factor Ndt80 is a key regulatory transition in the life cycle of Saccharomyces cerevisiae because it triggers exit from pachytene and entry into meiosis. The NDT80 promoter is held inactive by a complex containing the DNA-binding protein Sum1 and the histone deacetylase Hst1. Meiosis-specific phosphorylation of Sum1 by the protein kinases Cdk1, Ime2, and Cdc7 is required for NDT80 expression. Here, we show that the S-phase-promoting cyclin Clb5 activates Cdk1 to phosphorylate most, and perhaps all, of the 11 minimal cyclin-dependent kinase (CDK) phospho-consensus sites (S/T-P) in Sum1. Nine of these sites can individually promote modest levels of meiosis, yet these sites function in a quasiadditive manner to promote substantial levels of meiosis. Two Cdk1 sites and an Ime2 site individually promote high levels of meiosis, likely by preparing Sum1 for phosphorylation by Cdc7. Chromatin immunoprecipitation reveals that the phosphorylation sites are required for removal of Sum1 from the NDT80 promoter. We also find that Sum1, but not its partner protein Hst1, is required to repress NDT80 transcription. Thus, while the phosphorylation of Sum1 may lead to dissociation from DNA by influencing Hst1, it is the presence of Sum1 on DNA that determines whether NDT80 will be expressed.

Burden and predictors of hypertension in India: results of SEEK (Screening and Early Evaluation of Kidney Disease) study.

BACKGROUND: Hypertension (HTN) is one of the major causes of cardiovascular morbidity and mortality. The objective of the study was to investigate the burden and predictors of HTN in India. METHODS: 6120 subjects participated in the Screening and Early Evaluation of Kidney disease (SEEK), a community-based screening program in 53 camps in 13 representative geographic locations in India. Of these, 5929 had recorded blood pressure (BP) measurements. Potential predictors of HTN were collected using a structured questionnaire for SEEK study. RESULTS: HTN was observed in 43.5% of our cohort. After adjusting for center variation (p < 0.0001), predictors of a higher prevalence of HTN were older age ≥ 40 years (p < 0.0001), BMI of ≥ 23 Kg/M2 (p < 0.0004), larger waist circumference (p < 0.0001), working in sedentary occupation (p < 0.0001), having diabetes mellitus (p < 0.0001), having proteinuria (p < 0.0016), and increased serum creatinine (p < 0.0001). High school/some college education (p = 0.0016), versus less than 9th grade education, was related with lower prevalence of HTN. Of note, proteinuria and CKD were observed in 19% and 23.5% of HTN subjects. About half (54%) of the hypertensive subjects were aware of their hypertension status. CONCLUSIONS: HTN was common in this cohort from India. Older age, BMI ≥ 23 Kg/M2, waist circumference, sedentary occupation, education less, diabetes mellitus, presence of proteinuria, and raised serum creatinine were significant predictors of hypertension. Our data suggest that HTN is a major public health problem in India with low awareness, and requires aggressive community-based screening and education to improve health.

Nontraditional cardiovascular biomarkers and estimation of cardiovascular risk in predialysis chronic kidney disease patients and their correlations with carotid intima media thickness.

BACKGROUND: Cardiovascular biomarkers such as N-terminal pro-B-type natriuretic peptide (NT-proBNP), cardiac troponin T (cTnT), hs-CRP (high sensitivity C-reactive protein), and albuminuria predict underlying heart disease in the general population as well as CKD patients. OBJECTIVES: We aimed to study the association of NT-proBNP, cTnT, hs-CRP, and spot urine albumin creatinine ratio with carotid intima media thickness (CIMT) for cardiovascular risk estimation in predialysis CKD (chronic kidney disease) patients. PATIENTS AND METHODS: This cross-sectional study included a total of 120 adult predialysis CKD patients. Forty patients were allocated in each predialysis CKD group of stages 3, 4, and 5. Serum cTnT and hs-CRP, plasma NT-proBNP, and single spot urine albumin creatinine ratio (ACR) were measured. Ultrasonographic examination of carotid artery was done with 7.5 MHz linear probe in B mode ultrasonography and carotid intima media thickness was measured. RESULTS: Mean values ± standard deviation of plasma NT-proBNP (pg/mL), serum hs-CRP (mg/L), spot urine ACR (mg/g of creatinine), and CIMT (mm) were 585.68 ± 514.84, 5.96 ± 2.52, 719.37 ± 411.36, and 0.78 ± 0.15, respectively in predialysis CKD patients (n = 120). Serum cTnT level was high in 40% of predialysis CKD patients. Among cardiovascular biomarkers, plasma NT-proBNP had maximum strength of correlation (Spearman Rho correlation coefficient; r = 0.575 and P < 0.0001) with the carotid intima media thickness followed by serum cTnT (Spearman Rho correlation coefficient; r = 0.419 and P < 0.0001), spot urine albumin creatinine ratio (Spearman Rho correlation coefficient; r = 0.322 and P < 0.0001), and serum hs-CRP (Spearman Rho correlation coefficient; r = 0.246 and P = 0.007). CONCLUSIONS: Nontraditional cardiovascular biomarkers such as plasma NT-proBNP, serum cTnT, serum hs-CRP, and spot urine ACR significantly correlate with CIMT. These biomarkers can estimate the cardiovascular risk in a predialysis CKD population with expected high cardiac morbidity and mortality.