Revealing floral metabolite network in tuberose that underpins scent volatiles synthesis, storage and emission.

KEY MESSAGE: The role of central carbon metabolism in the synthesis and emission of scent volatiles in tuberose flowers was revealed through measurement of changes in transcripts and metabolites levels. Tuberose or Agave amica (Medikus) Thiede & Govaerts is a widely cultivated ornamental plant in several subtropical countries. Little is known about metabolite networking involved in biosynthesis of specialized metabolites utilizing primary metabolites. In this study, metabolite profiling and gene expression analyses were carried out from six stages of maturation throughout floral lifespan. Multivariate analysis indicated distinction between early and late maturation stages. Further, the roles of sugars viz. sucrose, glucose and fructose in synthesis, glycosylation and emission of floral scent volatiles were studied. Transcript levels of an ABC G family transporter (picked up from the floral transcriptome) was in synchronization with terpene volatiles emission during the anthesis stage. A diversion from phenylpropanoid/benzenoid to flavonoid metabolism was observed as flowers mature. Further, it was suggested that this metabolic shift could be mediated by isoforms of 4-Coumarate-CoA ligase along with Myb308 transcription factor. Maximum glycosylation of floral scent volatiles was shown to occur at the late mature stage when emission declined, facilitating both storage and export from the floral tissues. Thus, this study provides an insight into floral scent volatiles synthesis, storage and emission by measuring changes at transcripts and metabolites levels in tuberose throughout floral lifespan.

Phenotype Algorithm based Big Data Analytics for Cancer Diagnose.

Nowadays, Cancer diagnosis is one of the major challenging characteristics for treating cancer. The reality of cancer patients rely on the diagnosis of cancer at the early stages (either in stage 1 or stage 2). If the cancer is diagnosed in stage 3 or later stages means the changes of survival of the patient will become more critical. Normally, single patient records will generate a huge amount of data if the data could be manage and analyze means to solve many problems for identifying the patterns it will leads to diagnose the cancer. Recent work several machine learning algorithms are introduced for the classification of cancer. However still the classification accuracy of machine learning algorithms are reduced because of huge number of samples. So the proposed work introduces a new Hadoop Distributed File System (HDFS) is focused in this work. In this paper, the proposed phenotype techniques are used which handle and classifies the raw EHR (Electronic Health Record) and EMR (Electronic Medical Record). It is based on the HDFS and Two-Phase Map Reduce. Phenotype algorithm uses NLP (National Language Processing) tool which will analyze and classify the cancer patient data like gene mapping, age related data, image and ultrasonic frequency processing, identification and analysis of irregularities, disease and personal histories. In this paper, the three factorized model is used which calculates the mean score values. The values are calculated by disease stage, pain status, etc. This paper focuses big data analytics for cancer diagnosis and the simulation results shows the proposed system produces the highest performance.

Quorum sensing inhibitory activity of the metabolome from endophytic Kwoniella sp. PY016: characterization and hybrid model-based optimization.

Quorum sensing, the microbial communication system, is gaining importance as a therapeutic target against pathogens. The two key reasons for the rising demand of quorum sensing (QS) inhibitory molecules are low selective pressure to develop resistance by pathogens and possibility of more species-specific effects. Due to complex interactions in a unique niche of live plant tissues, endophytes, as a survival mechanism, potentially produce various bioactive compounds such as QS inhibitors. We report the isolation of an endophytic fungus Kwoniella sp. PY016 from the medicinal plant "Bahera" (Terminalia bellirica), which exhibits substantial quorum sensing inhibition and anti-biofilm activities against the standard test organism, Chromobacterium violaceum. Sugar, sugar alcohol, carboxylic acid, lipid, and phenolic classes of metabolites (predominantly xylitol) are responsible components of the metabolome for the desired bioactivity. A judicious combination of single-factor-at-a-time strategy and artificial neural network modeling combined with genetic algorithm was employed for the selection and optimization of the critical process and medium parameters. Through this newly adopted hybrid model-based optimization, the quorum sensing inhibitory activity of the endophytic metabolome was increased by ~ 30%. This is the first report on optimization of QS inhibitory activity from any fungal endophyte using such a hybrid advanced approach.

Mutational analyses of regulatory genes, mexR, nalC, nalD and mexZ of mexAB-oprM and mexXY operons, in efflux pump hyperexpressing multidrug-resistant clinical isolates of Pseudomonas aeruginosa.

The present study deals with membrane-bound efflux pumps, MexAB-OprM and MexXY and their respective regulatory genes mexR, nalC, nalD and mexZ in multidrug resistant (MDR) Pseudomonas aeruginosa. Following antibiotic sensitivity testing and detection of various beta-lactamases, hyperexpression of efflux pump genes, mexB and mexY in the isolates was investigated using semi-quantitative and real-time reverse transcription-PCR. Amplicons from regulatory genes were sequenced and subjected to mutational and phylogenetic analysis. Twenty-nine clinical isolates of P. aeruginosa were obtained from a total of 144 MDR gram-negative bacteria collected from Kerala State, South India. All strains were found to be resistant to ampicillin and nalidixic acid with 13.8, 44.8 and 31% testing positive for extended-spectrum beta-lactamases, metallo-beta-lactamases and AmpC producers respectively. Increased mexB and mexY transcription was detected respectively in 10.3 and 20.7% of the isolates in comparison with P. aeruginosa reference strain, PAO (MTCC). Co-expression of MexY was also observed in MexB overproducers. Various synonymous/and non-synonymous mutations in regulatory gene sequences of efflux pump operons were detected. In the strain designate Pa16, mexR was found to harbour four novel point mutations with one transversion and three transitions which included a substitution of an ochre codon with that for serine. The gene also displayed a novel mutation involving insertion of a cysteine at the 444th base position, followed by an opal codon. The genetic divergence and homogeneity of the concatenated (mexR, nalC and nalD) regulatory gene sequences of mexAB-oprM operon was apparent in the phylogram generated with similar sequences retrieved from public database.

Analysis of beta-lactamases, blaNDM-1phylogeny & plasmid replicons in multidrug-resistant Klebsiella spp. from a tertiary care centre in south India.

BACKGROUND & OBJECTIVES: ß-lactamases play a predominant role in drug-resistance amongst Enterobacteriaceae. Presence of genes on transferable plasmids encoding these enzymes favours their dissemination across species and genera within and outside geographical boundaries. This study was aimed to understand the presence of ß-lactamases and transferable plasmids in clinical isolates of Klebsiella spp. which can contribute to the spread of resistance determinants. METHODS: A total of 41 clinical isolates of Klebsiella spp., collected from a tertiary care centre in Kerala, India, were checked for antibiotic sensitivity and the presence of plasmids. The ability to produce extended-spectrum ß-lactamases (ESBLs) and metallo-ß-lactamases (MBLs) was screened for and confirmed in 29 plasmid-harbouring isolates. blaNDM-1-specific primers were used for polymerase chain reaction amplification with plasmid DNA as template to determine episomal prevalence of this gene and its sequence-based phylogeny employing similar sequences from GenBank. Plasmid replicon typing was also carried out to determine the presence of transferable plasmids. RESULTS: Our results showed a high degree of multidrug-resistant (MDR) pathogens with ESBL production confirmed in 52 per cent, MBL in 31 per cent and co-production of both enzymes in seven per cent of the plasmid-bearing isolates. Plasmid DNA from 14 per cent of the isolates produced blaNDM-1-specific amplicons which showed sequence homology with those from bacteria of different genera and geographical areas. The predominant replicon type was found to be that of conjugative plasmids belonging to the incompatibility group - IncFIIK. INTERPRETATION & CONCLUSIONS: This study provides insight into the predominance of various ß-lactamases and potent gene-disseminating agents in Klebsiella spp. and emphasizes the need for constant surveillance of these pathogens to determine appropriate treatment strategies.

A new multi-criteria decision making method for the selection of construction contractors using interval valued fuzzy set.

OBJECTIVE: This article introduces a novel approach called Digital Weighted Multi Criteria Decision Making (DWMCDM) that employs interval valued fuzzy sets to select the best contractor for building projects. The contractor is chosen based on the pre-qualification and bid evaluation phases. In the first phase, the distance between the actual and required skills of the significant criteria is determined, and it is then converted into digital weighted distances to identify the maximum number of criteria related to the specific project of each contractor. The second step ranks the best contractor based on the bid price and digital weighted distances. RESULTS: The suggested technique integrates the pre-qualification and bid review phases to address project award delays and other restrictions. Finally, a real-world application is addressed to demonstrate the applicability of the proposed approach to any type of interval valued fuzzy inputs.

Bioformulation of Bacillus proteolyticus MITWPUB1 and its biosurfactant to control the growth of phytopathogen Sclerotium rolfsii for the crop Brassica juncea var local, as a sustainable approach.

Bacillus proteolyticus MITWPUB1 is a potential producer of biosurfactants (BSs), and the organism is also found to be a producer of plant growth promoting traits, such as hydrogen cyanide and indole acetic acid (IAA), and a solubilizer of phosphate. The BSs were reportedly a blend of two classes, namely glycolipids and lipopeptides, as found by thin layer chromatography and Fourier-transform infrared spectroscopy analysis. Furthermore, semi-targeted metabolite profiling via liquid chromatography mass spectroscopy revealed the presence of phospholipids, lipopeptides, polyamines, IAA derivatives, and carotenoids. The BS showed dose-dependent antagonistic activity against Sclerotium rolfsii; scanning electron microscopy showed the effects of the BS on S. rolfsii in terms of mycelial deformations and reduced branching patterns. In vitro studies showed that the application of B. proteolyticus MITWPUB1 and its biosurfactant to seeds of Brassica juncea var local enhanced the seed germination rate. However, sawdust-carrier-based bioformulation with B. proteolyticus MITWPUB1 and its BS showed increased growth parameters for B. juncea var L. This study highlights a unique bioformulation combination that controls the growth of the phytopathogen S. rolfsii and enhances the plant growth of B. juncea var L. Bacillus proteolyticus MITWPUB1 was also shown for the first time to be a prominent BS producer with the ability to control the growth of the phytopathogen S. rolfsii.

cAMP-stimulated Cl- secretion is increased by glucocorticoids and inhibited by bumetanide in semicircular canal duct epithelium.

BACKGROUND: The vestibular system controls the ion composition of its luminal fluid through several epithelial cell transport mechanisms under hormonal regulation. The semicircular canal duct (SCCD) epithelium has been shown to secrete Cl- under ß2-adrenergic stimulation. In the current study, we sought to determine the ion transporters involved in Cl- secretion and whether secretion is regulated by PKA and glucocorticoids. RESULTS: Short circuit current (Isc) from rat SCCD epithelia demonstrated stimulation by forskolin (EC50: 0.8 µM), 8-Br-cAMP (EC50: 180 µM), 8-pCPT-cAMP (100 µM), IBMX (250 µM), and RO-20-1724 (100 µM). The PKA activator N6-BNZ-cAMP (0.1, 0.3 & 1 mM) also stimulated Isc. Partial inhibition of stimulated Isc individually by bumetanide (10 & 50 µM), and [(dihydroindenyl)oxy]alkanoic acid (DIOA, 100 µM) were additive and complete. Stimulated Isc was also partially inhibited by CFTRinh-172 (5 & 30 µM), flufenamic acid (5 µM) and diphenylamine-2,2'-dicarboxylic acid (DPC; 1 mM). Native canals of CFTR+/- mice showed a stimulation of Isc from isoproterenol and forskolin+IBMX but not in the presence of both bumetanide and DIOA, while canals from CFTR-/- mice had no responses. Nonetheless, CFTR-/- mice showed no difference from CFTR+/- mice in their ability to balance (rota-rod). Stimulated Isc was greater after chronic incubation (24 hr) with the glucocorticoids dexamethasone (0.1 & 0.3 µM), prednisolone (0.3, 1 & 3 µM), hydrocortisone (0.01, 0.1 & 1 µM), and corticosterone (0.1 & 1 µM) and mineralocorticoid aldosterone (1 µM). Steroid action was blocked by mifepristone but not by spironolactone, indicating all the steroids activated the glucocorticoid, but not mineralocorticoid, receptor. Expression of transcripts for CFTR; for KCC1, KCC3a, KCC3b and KCC4, but not KCC2; for NKCC1 but not NKCC2 and for WNK1 but only very low WNK4 was determined. CONCLUSIONS: These results are consistent with a model of Cl- secretion whereby Cl- is taken up across the basolateral membrane by a Na+-K+-2Cl- cotransporter (NKCC) and potentially another transporter, is secreted across the apical membrane via a Cl- channel, likely CFTR, and demonstrate the regulation of Cl- secretion by protein kinase A and glucocorticoids.

Dynamic distress calls: volatile info chemicals induce and regulate defense responses during herbivory.

Plants are continuously threatened by a plethora of biotic stresses caused by microbes, pathogens, and pests, which often act as the major constraint in crop productivity. To overcome such attacks, plants have evolved with an array of constitutive and induced defense mechanisms- morphological, biochemical, and molecular. Volatile organic compounds (VOCs) are a class of specialized metabolites that are naturally emitted by plants and play an important role in plant communication and signaling. During herbivory and mechanical damage, plants also emit an exclusive blend of volatiles often referred to as herbivore-induced plant volatiles (HIPVs). The composition of this unique aroma bouquet is dependent upon the plant species, developmental stage, environment, and herbivore species. HIPVs emitted from infested and non-infested plant parts can prime plant defense responses by various mechanisms such as redox, systemic and jasmonate signaling, activation of mitogen-activated protein (MAP) kinases, and transcription factors; mediate histone modifications; and can also modulate the interactions with natural enemies via direct and indirect mechanisms. These specific volatile cues mediate allelopathic interactions leading to altered transcription of defense-related genes, viz., proteinase inhibitors, amylase inhibitors in neighboring plants, and enhanced levels of defense-related secondary metabolites like terpenoids and phenolic compounds. These factors act as deterrents to feeding insects, attract parasitoids, and provoke behavioral changes in plants and their neighboring species. This review presents an overview of the plasticity identified in HIPVs and their role as regulators of plant defense in Solanaceous plants. The selective emission of green leaf volatiles (GLVs) including hexanal and its derivatives, terpenes, methyl salicylate, and methyl jasmonate (MeJa) inducing direct and indirect defense responses during an attack from phloem-sucking and leaf-chewing pests is discussed. Furthermore, we also focus on the recent developments in the field of metabolic engineering focused on modulation of the volatile bouquet to improve plant defenses.

Light Weight Deep Learning Algorithm for Voice Call Quality of Services (Qos) in Cellular Communication.

In this paper, a deep learning algorithm was proposed to ensure the voice call quality of the cellular communication networks. This proposed model was consecutively monitoring the voice data packets and ensuring the proper message between the transmitter and receiver. The phone sends its unique identification code to the station. The telephone and station maintain a constant radio connection and exchange packets from time to time. The phone can communicate with the station via analog protocol (NMT-450) or digital (DAMPS, GSM). Cellular networks may have base stations of different standards, which allow you to improve network performance and improve its coverage. Cellular networks are different operators connected to each other, as well as a fixed telephone network that allows subscribers of one operator to another to make calls from mobile phones to landlines and from landlines to mobiles. The simulation is conducted in Matlab against different performance metrics, that is, related to the quality of service metric. The results of the simulation show that the proposed method has a higher QoS rate than the existing method over an average of 97.35%.

Expression of epithelial calcium transport system in rat cochlea and vestibular labyrinth.

BACKGROUND: The low luminal Ca2+ concentration of mammalian endolymph in the inner ear is required for normal hearing and balance. We recently reported the expression of mRNA for a Ca2+-absorptive transport system in primary cultures of semicircular canal duct (SCCD) epithelium. RESULTS: We now identify this system in native vestibular and cochlear tissues by qRT-PCR, immunoblots and confocal immunolocalization. Transcripts were found and quantified for several isoforms of epithelial calcium channels (TRPV5, TRPV6), calcium buffer proteins (calbindin-D9K, calbindin-D28K), sodium-calcium exchangers (NCX1, NCX2, NCX3) and plasma membrane Ca2+-ATPase (PMCA1, PMCA2, PMCA3, and PMCA4) in native SCCD, cochlear lateral wall (LW) and stria vascularis (SV) of adult rat as well as Ca2+ channels in neonatal SCCD. All components were expressed except TRPV6 in SV and PMCA2 in SCCD. 1,25-(OH)2vitamin D3 (VitD) significantly up-regulated transcripts of TRPV5 in SCCD, calbindin-D9K in SCCD and LW, NCX2 in LW, while PMCA4 in SCCD and PMCA3 in LW were down-regulated. The expression of TRPV5 relative to TRPV6 was in the sequence SV > Neonatal SCCD > Adult SCCD > LW > primary culture SCCD. Expression of TRPV5 protein from primary culture of SCCD did not increase significantly when cells were incubated with VitD (1.2 times control; P > 0.05). Immunolocalization showed the distribution of TRPV5 and TRPV6. TRPV5 was found near the apical membrane of strial marginal cells and both TRPV5 and TRPV6 in outer and inner sulcus cells of the cochlea and in the SCCD of the vestibular system. CONCLUSIONS: These findings demonstrate for the first time the expression of a complete Ca2+ absorptive system in native cochlear and vestibular tissues. Regulation by vitamin D remains equivocal since the results support the regulation of this system at the transcript level but evidence for control of the TRPV5 channel protein was lacking.

Prevalence and risk factors for diabetic retinopathy: a population-based assessment from Theni District, south India.

AIMS: To estimate the prevalence of diabetic retinopathy (DR) and the possible risk factors associated with DR, in a population of south India. METHODS: A cross-sectional sample of subjects aged 30 years and older was selected using a cluster sampling technique from Theni district of Tamilnadu state. Eligible subjects were identified through a door-to-door survey and fasting blood glucose estimation. History of diabetes was elicited, and height, weight and blood pressure were measured for all subjects. Ocular examinations including visual acuity and anterior and posterior segment examinations were performed at preselected sites within clusters. RESULTS: Among the 25 969 persons screened for diabetes mellitus (DM), 2802 (10.8%) (95% CI 9.3 to 12.2%) were found to have DM. DR was detected in 298 (1.2%) of 25 969 subjects. The age-gender-adjusted prevalence of DR is 0.05% (95% CI 0.04 to 0.06%) for rural and 1.03% (95% CI 0.89 to 1.12%) for urban areas. The overall age-gender-cluster adjusted prevalence of DR was 0.74% (95% CI 0.66 to 0.83%). Diabetic retinopathy was present in 12.2% (95% CI 10.4 to 14.1%) of the DM population. CONCLUSION: Adequate training of ophthalmologists in treating DR and improvement in eye-care infrastructure are needed to tackle this major public health problem in India.

Profiling of volatile and non-volatile metabolites in Polianthes tuberosa L. flowers reveals intraspecific variation among cultivars.

Polianthes tuberosa L. (tuberose) is a widely cultivated ornamental crop in Asian countries. Different cultivars of tuberose have been developed through breeding programs in India. However, no reports on floral fragrance and metabolite contents of these cultivars are available. In this study, an attempt has been made to evaluate the levels of both volatile and non-volatile metabolites from seven different cultivars of P. tuberosa. Presence of benzenoids, phenylpropanoids, terpenoids, and few fatty acid derivatives as emitted, endogenous and glycosylated forms were revealed from the studied cultivars. Further, chemometric analyses in both supervised and unsupervised manner led to identification of patterns among the cultivars. Among the seven cultivars, four distinct clusters were obtained linking to their volatiles, flavonoids and primary metabolite levels. Metabolic variations obtained from the cultivars also suggest cross-talks between phenylpropanoid, benzenoid, and flavonoid pathways. Thus metabolite profiling reported here may help in characterization of tuberose cultivars for perfumery utility and future breeding programme.

Developmental variation in floral volatiles composition of a fragrant orchid Zygopetalum maculatum (Kunth) Garay.

Emitted scent volatile profile of an orchid species Zygopetalum maculatum was studied using dynamic headspace sampling technique with four different adsorbent matrices, namely Porapak Type Q polymer (mesh size: 80/100), Tenax (mesh size: 60/80), activated charcoal and graphite. In addition, developmental variations in scent emissions and endogenous volatile levels were also investigated. Gas chromatography-mass spectrometry analysis revealed the presence of 21 volatile compounds in the headspace, which was predominantly enriched with benzenoid compounds. Among these benzenoids, o-diethylbenzene and p-diethylbenzene were the major compounds followed by benzyl acetate and methyl salicylate. Among the phenylpropanoid compounds, 2-phenylethyl acetate was the major volatile. However, as compared to benzenoids, the quantity was much lesser, indicating the inclination of phenylalanine flux towards benzenoid pathway. The outcome of this study has the implications in enhancing fragrance and vase life of orchids of the Sikkim Himalaya region and thus may further help to meet the growing market demand.

Structural, morphological, optical and biological properties of pure ZnO and agar/zinc oxide nanocomposites.

In this work, ZnO nanoparticles were prepared by in situ chemical precipitation method in the presence of Agar biopolymer. The influence of Agar concentrations on the structural, morphological and optical properties of ZnO have been investigated. The XRD pattern of Pure ZnO and Agar/ZnO nanocomposites indicates the hexagonal wurtzite phase of ZnO. The crystallite size of pure ZnO and Agar/ZnO nanocomposites was found to be in the range of 35.5 to 19.73 nm. Pure ZnO and Agar/ZnO nanocomposites showed nanospheroid and nanopaddy shaped morphology from FESEM studies. The interplanar distance observed from the HRTEM image confirms the plane of the prepared material. The elemental composition of the samples were characterized by EDX. The optical properties of Pure ZnO and Agar/ZnO nanocomposites were characterized by UV, FTIR and PL. The band gap of Agar/ZnO nanocomposites were varied with the Agar concentration. Oxygen vacancy induced photoluminescence of ZnO are observed and its intensity is found to be increased linearly with the Agar concentration. The antibacterial activity of ZnO and Agar/ZnO nanocomposites was evaluated by disc diffusion method against Gram-positive (B.subtilis) and Gram-negative (P. aeruginosa) bacteria. The cytotoxicity of Agar/ZnO nanocomposites was studied against Normal (L929) and Breast cancer cell line (MB231). The result of this investigation reveals that the Agar/ZnO nanocomposites deliver a dose dependent toxicity in normal and cancer cell line.

Pre-tertiary hospital care of patients with chronic kidney disease in India.

BACKGROUND & OBJECTIVES: There is paucity of data available on how chronic kidney disease (CKD) is treated before referral to a tertiary hospital. This study was conducted to assess pre-tertiary hospital care of patients with CKD 5 at their presentation to nephrology services at a tertiary care hospital. METHODS: Over a period of 8 months, consecutive patients with CKD 5 presenting at the Nephrology services at Christian Medical College, Vellore, Tamil Nadu, and their relatives were interviewed to assess the pre-tertiary hospital care and knowledge about CKD 5 and its treatment. RESULTS: A total of 561 patients with CKD 5 were enrolled. The mean duration (months) of known CKD was 12.4 +/- 23.1 and known CKD 5 was 3.2 +/- 3.5. Of these, 369 patients (65.8%) had been under the care of a nephrologist; 305 patients had CKD 5 as the initial presentation of renal illness. Vaccination against hepatitis B had been initiated in only 133 patients (23.7%). Only 172 patients(38%) had an adequately controlled blood pressure. Care under a nephrologist was more likely to result in appropriate investigation, treatment and patient education though blood pressure control did not differ. INTERPRETATION & CONCLUSION: Paucity of symptoms in the initial stages of certain forms of CKD probably led to 50 per cent of patients presenting with CKD 5 as the initial presentation of renal disease. Inadequate vaccination against hepatitis B infection highlights the need for appropriate vaccination. Prevention of CKD and its progression are important targets which requires physician awareness at all levels. Early referral to a nephrologist's care is more likely to result in appropriate investigations and treatment.

Glucocorticoid regulation of genes in the amiloride-sensitive sodium transport pathway by semicircular canal duct epithelium of neonatal rat.

The lumen of the inner ear has an unusually low concentration of endolymphatic Na+, which is important for transduction processes. We have recently shown that glucocorticoid receptors (GR) stimulate absorption of Na+ by semicircular canal duct (SCCD) epithelia. In the present study, we sought to determine the presence of genes involved in the control of the amiloride-sensitive Na+ transport pathway in rat SCCD epithelia and whether their level of expression was regulated by glucocorticoids using quantitative real-time RT-PCR. Transcripts were present for alpha-, beta-, and gamma-subunits of the epithelial sodium channel (ENaC); the alpha1-, alpha3-, beta1-, and beta3-isoforms of Na+-K+-ATPase; inwardly rectifying potassium channels [IC50 of short circuit current (Isc) for Ba2+: 210 microM] Kir2.1, Kir2.2, Kir2.3, Kir2.4, Kir3.1, Kir3.3, Kir4.1, Kir4.2, Kir5.1, and Kir7.1; sulfonyl urea receptor 1 (SUR1); GR; mineralocorticoid receptor (MR); 11beta-hydroxysteroid dehydrogenase (11beta-HSD) types 1 and 2; serum- and glucocorticoid-regulated kinase 1 (Sgk1); and neural precursor cell-expressed developmentally downregulated 4-2 (Nedd4-2). On the other hand, transcripts for the alpha4-subunit of Na+-K+-ATPase, Kir1.1, Kir3.2, Kir3.4, Kir6.1, Kir6.2, and SUR2 were found to be absent, and Isc was not inhibited by glibenclamide. Dexamethasone (100 nM for 24 h) not only upregulated the transcript expression of alpha-ENaC (approximately 4-fold), beta2-subunit (approximately 2-fold) and beta3-subunit (approximately 8-fold) of Na+-K+-ATPase, Kir2.1 (approximately 5-fold), Kir2.2 (approximately 9-fold), Kir2.4 (approximately 3-fold), Kir3.1 (approximately 3- fold), Kir3.3 (approximately 2-fold), Kir4.2 (approximately 3-fold), Kir7.1 (approximately 2-fold), Sgk1 (approximately 4-fold), and Nedd4-2 (approximately 2-fold) but also downregulated GR (approximately 3-fold) and 11beta-HSD1 (approximately 2-fold). Expression of GR and 11beta-HSD1 was higher than MR and 11beta-HSD2 in the absence of dexamethasone. Dexamethasone altered transcript expression levels (alpha-ENaC and Sgk1) by activation of GR but not MR. Proteins were present for the alpha-, beta-, and gamma-subunits of ENaC and Sgk1, and expression of alpha- and gamma-ENaC was upregulated by dexamethasone. These findings are consistent with the genomic stimulation by glucocorticoids of Na+ absorption by SCCD and provide an understanding of the therapeutic action of glucocorticoids in the treatment of Meniere's disease.

Assessment of cell cycle phase-specific effects of zerumbone on mitotically synchronous surface cultures of Physarum polycephalum.

Zerumbone, a natural cyclic sesquiterpene, has been the focus of recent research as it has been found to exhibit selective toxicity towards cancer cells compared to normal cells. Studies on the cell cycle phase-specific effects of this interesting compound, however, remain sparse. Hence, concentration and time-dependent effects of zerumbone were evaluated employing a suitable model system, the naturally synchronous surface cultures of Physarum polycephalum. Zerumbone treatment in S, early, and late G2 phases resulted in G2 arrest. Early G2 phase exhibited the highest sensitivity (P < 0.001) to the compound. Protein profiles showed a complete inhibition of cyclin B1 expression following zerumbone treatment. Furthermore, FACS and comet analysis revealed that zerumbone inhibited DNA synthesis (P < 0.001) without being genotoxic at the concentrations tested. Differential display of mRNA showed distinct zerumbone-induced variations in transcript profiles, an analysis of which suggested a likely link between cellular networks involving stress-related gene expression and G2 arrest in P. polycephalum.

Ion transport regulation by P2Y receptors, protein kinase C and phosphatidylinositol 3-kinase within the semicircular canal duct epithelium.

BACKGROUND: The ionic composition of the luminal fluid in the vestibular labyrinth is maintained within tight limits by the many types of epithelial cells bounding the lumen. Regulatory mechanisms include systemic, paracrine and autocrine hormones along with their associated intracellular signal pathways. The epithelium lining the semicircular canal duct (SCCD) is a tissue that is known to absorb sodium and calcium and to secrete chloride. FINDINGS: Transport function was assessed by measurements of short circuit current (Isc) and gene transcript expression was evaluated by microarray. Neither ATP nor UTP (100 microM) on the apical side of the epithelium had any effect on Isc. By contrast, basolateral ATP transiently increased Isc and transepithelial resistance dropped significantly after basolateral ATP and UTP. P2Y2 was the sole UTP-sensitive purinergic receptor expressed. Isc was reduced by 42%, 50% and 63% after knockdown of alpha-ENaC, stimulation of PKC and inhibition of PI3-K, while the latter two increased the transepithelial resistance. PKCdelta, PKCgamma and PI3-K were found to be expressed. CONCLUSIONS: These observations demonstrate that ion transport by the SCCD is regulated by P2Y2 purinergic receptors on the basolateral membrane that may respond to systemic or local agonists, such as ATP and/or UTP. The sodium absorption from endolymph mediated by ENaC in SCCD is regulated by signal pathways that include the kinases PKC and PI3-K. These three newly-identified regulatory components may prove to be valuable drug targets in the control of pathologic vestibular conditions involving dysfunction of transport homeostasis in the ear, such as Meniere's disease.

In vitro evaluation of antioxidants of fruit extract of Momordica charantia L. on fibroblasts and keratinocytes.

The antioxidant activity of the total aqueous extract (TAE) and total phenolic extract (TPE) of Momordica charantia fruits was assayed by radical-scavenging methods and cytoprotective effects on hydrogen peroxide (H(2)O(2))- and hypoxanthin-xanthin oxidase (HX-XO)-induced damage to rat cardiac fibroblasts (RCFs), NIH 3T3, and keratinocyte (A431). Cell viability was monitored by a 3-[4,5-dimethyltriazol-2-yl]-2,5-diphenyltretrazolium (MTT) assay. For fibroblasts, TPE at 200 and 300 microg/mL showed maximum and consistent cytoprotection against oxidants. The extract at 50 microg/mL also had significant and slightly protective effects on fibroblasts against H(2)O(2)- and HX-XO-induced damage, respectively. RCF was more tolerant toward the damage. For keratinocytes, a dose-dependent relationship of oxidant toxicity was only seen with H(2)O(2) but the protective action of the extract correlated with oxidant dosage. At 200 and 300 microg/mL TPE, cytoprotection was dose-dependent against oxidants. Extracts had no effect on HX-XO toxicity at 50 microg/mL. Pretreatment with both the extracts did not show any cytoprotection.