DET1 and COP1 Modulate the Coordination of Growth and Immunity in Response to Key Seasonal Signals in Arabidopsis

This is the final version. Available from Elsevier via the DOI in this record.Plant growth and development and outcomes of plant-microbe interactions are defined by coordinated responses to seasonal signals. The mechanisms that control the coordinated regulation of growth and immunity are not well understood. Here, we show that a common signaling module integrates environmental signals, such as photoperiod and temperature, to regulate the growth-defense balance. Key light-signaling components De-Etiolated 1 (DET1) and Constitutive Photomorphogenic 1 (COP1) negatively regulate immunity and are essential for immune modulation by photoperiod and temperature. Our results show that this is regulated by the transcription factor Phytochrome Interacting Factor 4 (PIF4), suggesting that the DET1/COP1-PIF4 module acts as a central hub for the control of growth and immunity in response to seasonal signals. These findings provide a regulatory framework for environmental signal integration. In plants, adaptive traits such as growth and immunity are strongly influenced by the environment. How multiple seasonal signals are integrated is not well understood. Gangappa and Kumar show that a common signaling module comprising DET1, COP1, and PIF4 coordinates growth and immunity in response to key seasonal signals

DET1 and HY5 Control PIF4-Mediated Thermosensory Elongation Growth through Distinct Mechanisms

This is the final version. Available from Elsevier via the DOI in this record.Plant growth and development are defined by environmental cues. The transcription factor PHYTOCHROME INTERACTING FACTOR 4 (PIF4) is the central signaling hub that integrates environmental cues, including light and temperature, to regulate growth and development. The thermosensory mechanisms controlling the PIF4-mediated temperature response, and its integration with other environmental responses, remain poorly understood. DE-ETIOLATED 1 (DET1) and CONSTITUTIVE PHOTOMORPHOGENESIS 1 (COP1), key regulators of light signaling, have been proposed to control thermosensory growth by transcriptional regulation of PIF4, through ELONGATED HYPOCOTYL 5 (HY5). Here, we show that DET1/COP1 and HY5 regulate thermosensory elongation through distinct mechanisms. DET1 and COP1 are essential for promoting PIF4 expression and stabilizing PIF4 protein. Furthermore, HY5 inhibits elongation growth through competitive chromatin binding to PIF4 targets, not through transcriptional regulation of PIF4. Our findings reveal a mechanistic framework in which DET1/COP1 and HY5 regulatory modules act independently to regulate growth through the environmental signal integrator PIF4.Biotechnology and Biological Sciences Research Counci

SWR1 Chromatin-Remodeling Complex Subunits and H2A.Z Have Non-overlapping Functions in Immunity and Gene Regulation in Arabidopsis

This is the final version. Available from Elsevier via the DOI in this record.Incorporation of the histone variant H2A.Z into nucleosomes by the SWR1 chromatin remodeling complex is a critical step in eukaryotic gene regulation. In Arabidopsis, SWR1c and H2A.Z have been shown to control gene expression underlying development and environmental responses. Although they have been implicated in defense, the specific roles of the complex subunits and H2A.Z in immunity are not well understood. In this study, we analyzed the roles of the SWR1c subunits, PHOTOPERIOD-INDEPENDENT EARLY FLOWERING1 (PIE1), ACTIN-RELATED PROTEIN6 (ARP6), and SWR1 COMPLEX 6 (SWC6), as well as H2A.Z, in defense and gene regulation. We found that SWR1c components play different roles in resistance to different pathogens. Loss of PIE1 and SWC6 function as well as depletion of H2A.Z led to reduced basal resistance, while loss of ARP6 fucntion resulted in enhanced resistance. We found that mutations in PIE1 and SWC6 resulted in impaired effector-triggered immunity. Mutation in SWR1c components and H2A.Z also resulted in compromised jasmonic acid/ethylene-mediated immunity. Genome-wide expression analyses similarly reveal distinct roles for H2A.Z and SWR1c components in gene regulation, and suggest a potential role for PIE1 in the regulation of the cross talk between defense signaling pathways. Our data show that although they are part of the same complex, Arabidopsis SWR1c components could have non-redundant functions in plant immunity and gene regulation

Unusual formation and sub-omohyoid course of external jugular vein.

Variations in the origin and termination of external jugular vein are common and are reported in the past. However, variations in the course of external jugular vein are uncommon. During routine dissection classes for medical undergraduates, we came across the unusual formation and course of right external jugular vein and absence of common facial vein, in an approximately 60-year-old male cadaver of Indian origin. External jugular vein was formed by the continuation of undivided trunk of retromandibular vein. Following its formation, it passed vertically superficial to sternocleidomastoid muscle to the lower part of occipital triangle. In the occipital triangle it pierced the investing layer of deep cervical fascia and passed deep to the inferior belly of omohyoid muscle and coursed through the subclavian triangle. Then, it terminated at the junction of subclavian vein with internal jugular vein. Facial vein joined with submental vein and finally drained into internal jugular vein. Further, the posterior auricular vein and anterior jugular veins were absent. Knowledge about the variations of the retromandibular vein, common facial vein and external jugular vein observed in this study, may be important for the surgeons, to prevent inadvertent injury and excessive bleeding during diagnostic and therapeutic procedures

Development of Mucoadhesive Nanoparticulate System of Ebastine for Nasal Drug Delivery

Purpose: To prepare and evaluate mucoadhesive nanoparticulate system of ebastine for nasal drug delivery.Methods: The nanoparticles were prepared by ionic gelation method using drug-chitosan weight ratios 1:1, 1:2 and 1:3, and incorporating 0.5 or 0.7 % w/v sodium tripolyphosphate (STPP) and poloxamer 407. The mucoadhesive nanoparticles were characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), differential scanning colorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR) and evaluated for drug loading, entrapment efficiency, in vitro mucoadhesion, in vitro drug release and ex-vivo permeation.Results: FTIR and DSC studies indicate that no chemical interaction occurred between the drug and polymer. Nanoparticle size ranged from 169 to 500 nm. Drug loading and entrapment efficiency increased with increase in chitosan concentration and decreased with increase in poloxamer 407 concentration. The highest drug loading obtained for the nanoparticles was 19.5 %. With increase in polymer (chitosan) concentration (1:1 to 1:3), production yield was unchanged (73.2 to 74.4 % (F6)). Mucoadhesion increased with increase in the concentration of chitosan. In vitro drug release from all the formulations was biphasic, being characterized by a slight ‘burst’ followed by slow release. At the end of 8 h F6 (1:3) showed drug release of only 86.9 %, indicating sustained release. Ex-vivo permeation of pure ebastine was more rapid than from F6, thus indicating the capability of chitosan to control drug permeation rate through sheep nasal mucosa.Conclusion: The results indicate that a mucoadhesive nanoparticulate system can be used effectively for the nasal delivery of the antihistamine, ebastine.Keywords: Chitosan, Ebastine, Mucoadhesive, Nanoparticles, Ionotropic gelation, Permeation, Drug release, Poloxame

Isolation and identification of microsatellite repeat motifs from the Epinephelus fuscoguttatus genome

Epinephelus fuscoguttatus belongs to one of the largest serranidae fish family. Genetic information regarding existing fish populations in the wild is crucial for the conservation, particularly since the species is listed under the IUCD Red List due to intense fishing. Microsatellites of E. fuscoguttatus were isolated using streptavidin-biotin enrichment method. In total, 378 microsatellites were identified and characterized. Of these 378 total microsatellites, 46 (12.2%) were mononucleotides, 175 (46.3%) were dinucleotides, 109 (28.8%) were trinucleotides, 36 (9.5%) were tetranucloetides, 7 (1.9%) were pentanucleotides, 4 (1.1%) were hexanucleotides and 1 (0.3%) was a heptanucleotide. The most abundant microsatellite present in E. fuscoguttatus was the dinucleotide motif, (AC)n.Key words: Enrichment, Epinephelus fuscoguttatus, microsatellite, diversity

Congenital tuberculosis with multisystem involvement

Congenital tuberculosis is an uncommon condition, but in a country like India, where largest tuberculosis burden is found, there should be an eye on early diagnosis and management of this condition so as to prevent the devastating consequences of this disease considering the absence of specific pathogenic signs and symptoms. Here, we present a case of congenital tuberculosis in a newborn who was admitted on the 17th day after birth with the complaint of difficulty in breathing and lethargy for 3 days. On observing the antenatal history, it was found that mother after conception was diagnosed with pulmonary tuberculosis and was inadequately treated. The clinical indexes included no increase in body weight, hepatosplenomegaly, lethargy, and emaciated appearance. The treatment was initiated with antibiotics in intravenous form along with 4-drug antitubercular treatment (ATT). The 4-drug ATT was stopped after 1 week due to negative outcome in the tubercular screening, which leads to respiratory complications. The 4-drug ATT regimen was started again after a week on clinical basis which resulted in significant improvement within a few days. Hence, the present case presents a perspective that congenital tuberculosis can be considered in a newborn when mother has a history of tuberculosis before or during pregnancy

STABILITY INDICATING CHIRAL HPLC METHOD FOR THE ESTIMATION OF PIOGLITAZONE ENANTIOMERS IN PHARMACEUTICAL FORMULATION

Objective: A stability indicating chiral high performance liquid chromatographic (HPLC) method was developed and validated for the separated (S)and (R) pioglitazone in raw material and its determination in the presence of degradation products formed during forced degradation studies.Methods: In the present study, an isocratic normal phase-HPLC method was developed with stationary phase as ACI Cellu 1 (150 mm × 4.6 mm i.d.,5  μ)  column  and  n-hexane:  N-propyl  alcohol  (80:20,  V/V)  as  mobile  phase.  The  entire  study  was  performed  using  1.0  ml/minute  as  flow  rate  and the detection wavelength at  233 nm. The pioglitazone (R and S) was exposed to various stress condition such as hydrolytic (acid and base), neutral, oxidative, and photolytic. The stressed samples were analyzed by the proposed method.Result:  The  described  method  was  linear  over  the  range  of  5-15 µg/ml  for  R-pioglitazone  and  4-14 µg/ml  for  S-pioglitazone.  The  limit  of  detection and limit of quantification of S-pioglitazone and R-pioglitazone were found to be 1.4 μg/ml and 4.26 μg/ml, respectively. The recovery study of S and R-Pioglitazone from tablets formulation ranged from 97.14% to 100.04%, respectively.Conclusion: The developed method can be applied in the quality control of drug products.Keywords: Stability-indicating method, Validation, Chiral, Pioglitazone

Colorimetric Determination of Cefadroxil and Ceftriazone in Pharmaceutical Dosage Forms

Purpose: To develop a simple, rapid and selective method for the spectrophotometric determination of cefadroxil and ceftriaxone using 1, 2- napthaquinone-4- sulfonic acid sodium.Methods: The method was based on the derivatization of cefadroxil and ceftriaxone with 1, 2-naphthaquinone-4- sulfonic acid sodium in alkaline medium to yield orange-colored products.Results: The reaction products of cefadroxil and ceftriaxone at their respective max of 475 and 480 nm showed linearity in the concentration range of 10 - 100 and 25 - 175 ìg/ml, respectively. Relative standard deviations of 0.82 % for cefadroxil and 0.95 % for ceftriaxone were obtained. Recoveries of cefadroxil tablets and ceftriaxone injection were in the range of 100.66 ± 0.98 and 99.38 ± 0.84 %, respectively.Conclusion: Recovery studies gave satisfactory results indicating that none of the major additives/excipients interfered with the assay method. Therefore, the proposed method is simple, rapid, precise and convenient for the assay of cefadroxil and ceftriaxone in commercial preparations

In Vitro Antioxidant and In Vivo Hepatoprotective Activity of Leave Extract of Raphanus Sativus in Rats Using CCL4 Model

Background: Raphanus sativus is reported to have a variety of biological activities. This work screened the hepato-protective and antioxidant activity of ethanol (ERS), and aqueous (ARS), extracts of leaves of Raphanus sativus in Carbon tetrachloride (CCl4), model in rats.Material and Methods: The extracts were subjected to antioxidant tests (Total reducing power and Total phenolic content), and preliminary phytochemical screening. A pilot study was done on 100 and 300 mg/kg extracts, form which 300 mg was chosen for further experiments. The albino rats (200-250 grams), were divided into 5 groups of 6 animals each (n=6). There were three control groups comprising of normal control (normal saline -1ml/kg), negative control group (CCl4 1ml/kg in olive oil in a ratio of 1:1 v/v), and positive control group (Silymarin 50mg/kg). The Test drugs were given in a dose of 300 mg/kg for both ERS and ARS extract for 7 days. Biochemical parameters (AST, ALT, Alkaline phosphatase, Total Bilirubin), histo-pathological examination of liver and in vivo antioxidant tests [CAT, GSH and MDA] were done.Results: The phytochemical study showed the presence of flavanoids, terpenoids, alkaloids, saponins and sterols. A dose dependent increase in the oxidative potential was observed in both the extracts with total phenolic content 70.1 and 44.4 GAE/g extract for ERS and ARS respectively. ERS 300mg/kg showed a significant (p<0.001) increase in levels of AST, ALT and alkaline phosphatase as compared to negative control (percentage hepatoprotection =45.3%) while ARS 300 mg/kg (p<.01) group showed 30% hepatoprotection. The GSH (p<0.001) and CAT (p<0.05) in ERS and ARS were significantly increased while MDA levels were decreased (P< 0.01), as compared negative control. The findings were confirmed histo-pathological examination.Conclusion: The ethanol and aqueous extract of Raphanus sativus have partial hepatoprotection against CCl4 toxicity.Key words: Raphanus sativus, hepatoprotection, CCl4, antioxidant