Convergence of RAS and PP2A activities on chromatin repressor complexes

Protein Phosphatase 2A (PP2A) is a serine-threonine phosphatase and, through its de-phosphorylation activity, mostly downregulates signaling pathways, while RAS belongs to the family of small GTPase and activates various kinases resulting in the activation of cellular signaling. Decades back, it was shown that the inhibition of PP2A is a prerequisite for human cell transformation by the oncogenic RAS. Despite the mounting evidence highlighting RAS and PP2A-mediated protein phosphorylation to be of paramount significance in cancer biology, none of the studies have systematically addressed the cooperation between RAS and PP2A in regulating global protein phosphorylation. My thesis focuses on the cooperation between the tumor suppressor PP2A and an oncogene RAS in regulating protein phosphorylation in cancer cells. By integrating the RAS- and PP2A-regulated phosphoproteomics data we found that the cellular proteome, is specifically enriched for RAS- and PP2A-regulated phosphosites on proteins involved in regulating various epigenetic modifications such as DNA methylation, histone methylation/acetylation and chromatin remodeling. In the present work, I have validated the impact of the PP2A- and RAS-regulated phosphosites in the functional regulation of epigenetic proteins. Using multi-omics analysis, this study has further uncovered the opposing roles of RAS and PP2A in regulating gene expression. The present study has also compared the two nuclear inhibitors of PP2A, PME-1, and SET, and discovered the diversity of their roles in oncogenic transcription and DNA methylation. Collectively, this research work indicates that in cancer, RAS hyperactivity and PP2A inhibition converge on phosphorylation of epigenetic proteins and has a vital role in oncogenic transcription.RAS onkoproteiinin ja PP2A kasvunestäjän yhteisvaikutus geeninluentaan syövässä Proteiinifosfataasi 2A (PP2A) on seriini-treoniinifosfataasi, joka vaimentaa monien syövän kannalta tärkeiden signalointireittien aktiivisuutta, kun taas RAS, joka kuuluu pienien GTPaasi proteiinien perheeseen, aktivoi solusignalointikinaaseja syövässä. Samanaikainen RAS:in aktivaatio ja PP2A:n inhibitio on edellytyksenä ihmisen solujen muuttumiselle pahanlaatuisiksi. Kuitenkaan tähän mennessä ei ole systemaattisesti tutkittu RAS:in ja PP2A:n yhteistyötä proteiinien fosforylaation säätelyssä syöpäsoluissa. Tämä onkin ollut väitöskirjatutkimukseni keskeinen kysymys. Yhdistämällä proteomiikka-analyysit RAS- ja PP2A-muokatuista syöpäsoluista havaitsimme, että RAS:in ja PP2A:n säätelemät fosforylaatiokohdat esiintyvät rikastuneina proteiineissa, jotka osallistuvat erilaisten epigeneettisten modifikaatioiden kuten DNA:n metylaation, histonien metylaation/asetylaation sekä kromatiinin uudelleenmuokkauksen säätelyyn. Osoitan tässä työssä, että nämä fosforylaatiokohdat vaikuttavat epigeneettisten proteiinien toiminnan säätelyyn. Tutkimuksessa havaittiin myös, että RAS:lla ja PP2A:lla on erilainen rooli geenien ilmentymisen säätelyssä. Havaitsin myös, että kahdella PP2A:ta estävällä tumaproteiinilla, PME-1 ja SET, on erilainen rooli syöpään liittyvien geenien luennassa sekä DNA:n metylaation säätelyssä. Kaiken kaikkiaan väitöskirjatutkimukseni osoittaa, että RAS:in yliaktiivisuuden ja PP2A:n eston vaikutukset kohtaavat epigeneettisten proteiinien fosforylaation säätelyssä ja että tällä on tärkeä rooli onkogeenisessä geeninluennassa

Performance Comparison of Symmetric and Offset Reflector Antennas Adaptively Illuminated by Novel Triple Mode Feedhorn

Parabolic symmetric and offset reflector antennas adaptively illuminated using a novel triple-mode feedhorn (TE11+TM01+TE21) with different mode combinations and impedance and radiation performances are presented. The combination of the radiating modes in a feedhorn with proper amplitude and fixed phase values helps in electronically pointing the main beam of the radiating patterns such as that obtained in a beam-steering antenna with limited beam-scan range. This type of radiation performance virtually creates a displaced phase center location for the feedhorn, which, consequently, adaptively illuminates the reflector antenna surface. Impedance-matching bandwidths are preserved for both reflector antennas similar to the case of feedhorn alone. The copolarization gain and peak cross-polarization levels are far better with the offset reflector antenna than the symmetric reflector antenna. Such reflector antennas find applications in ground moving target indicator (GMTI) and space based radars. The investigation results are solely computed using FEKO full-wave analysis tool

Analysis of accuracy of burette in determination of surface tension of liquids and study of its variation with detachment time and inclination angle

1676-1684A simple laboratory experiment to measure the surface tension of liquids has been proposed in this work, which is well suited for any undergraduate laboratory demonstration. Burette with modified Tate’s law has been found to return surface tension values as good as those obtained using Traube’s stalagmometer. Experimental data and error analysis show similar result accuracy. Also, a comparative study brings to notice the exactitude of drop weight method over drop number method, both from stalagmometer and burette. The rate of flow of 11–23 drops per minute has been found to be highly suitable for minimization of errors due to evaporation and satellite drop formation. Angular dependence of surface tension has been proposed numerically and validated for the respective case with the angle of 0° to deviate the least from literature value. The results obtained in this study are synchronous with the established theoretical understanding of the concept

PHARMACEUTICAL AND ANALYTICAL STANDARDIZATION OF MAHA SHANKHA VATI

Khalveeya Rasa is the combinations of herbal, mineral and animal products, so that we can have the effects of all collectively in a single formula. It preserve the properties of freshly added Churna, Swarasa etc., with the help of Moorchita Parada i.e., Kajjali, Rasashindura & Hingula etc. Because of which Khalveeya Rasaushadhies occupies greater portion in therapeutics as compare to other Kalpana, Such as Vati, Gutika, Taila, Ghrita. In the present study three sample of “Maha Shankha Vati†have been prepared by adopting method describe in Ayurvedic Formulary of India (A.F.I.) Vol. 2 approved by government of India with some desire changes.AIMS: The aim of this study is to prepare three sample of “Maha Shankha Vati†for its Pharmacoanalytical standardization. And also prepare “Shankha Bhasmaâ€, “Hingullotha Parada†and processing of last three Sanskara of Parada. Analytical Standardization of self made Maha Shankha Vati & market sample of Maha Sankha Vati to compare analytically.RESULT: After evaluating organoleptic characters of the first three samples of self made Maha Shankha Vati revealed black colored substance in the form of Vati having Amla-Katu Rasa, Amla smell and smooth touch having pH 3.39. Similarly in case of market sample (fourth sample) shows black in colored in the form of Vati with Lavan-Katu in Rasa, Amla Smell and smooth touch having pH 4.37. Further the XRD pattern analysis of the combined sample of Maha Shankha Vati, reveals the presence of HgS, Caco3, NaCl. Same observation found in market sample.

New Material of Carbonaceous Compressions from the ∼1.5 Ga Singhora Group, Chhattisgarh Supergroup, India, and their Interpretation as Benthic Algae

The origin, antiquity, and affinity of benthic seaweeds (multicellular algae) in the geological past are shrouded in mystery due to their preservation bias. In this study, we present a new material of well-preserved carbonaceous compression fossils in shale horizons of the Mesoproterozoic (ca. ∼1,500–1,300 Ma) Singhora Group of the Chhattisgarh Supergroup. Eleven distinct taxa, including one new taxon, Palaeoscytosiphon shuklaii, n. gen. et. sp., and one new species, Jiuqunaoella sergeevii, n. sp., are established. Four unidentified morphologies are also reported. Morphologically, the carbonaceous fossils are fan-shaped, palmate, elongated, leaf-like algal thalli with/without holdfast at the base, isolated or dichotomously branched long filaments, along with multicellular reproductive structures. The results of laser Raman spectroscopy and energy dispersive X-ray spectroscopy (EDX) are also presented in support of their biogenicity. Collectively, the preservation mode of the Singhora carbonaceous fossils suggests multicellular algal affinity and adds to a Burgess Shale-type (BST) taphonomic window in the Pre-Ediacaran biosphere

Analysis of accuracy of burette in determination of surface tension of liquids and study of its variation with detachment time and inclination angle

A simple laboratory experiment to measure the surface tension of liquids has been proposed in this work, which is well suited for any undergraduate laboratory demonstration. Burette with modified Tate’s law has been found to return surface tension values as good as those obtained using Traube’s stalagmometer. Experimental data and error analysis show similar result accuracy. Also, a comparative study brings to notice the exactitude of drop weight method over drop number method, both from stalagmometer and burette. The rate of flow of 11–23 drops per minute has been found to be highly suitable for minimization of errors due to evaporation and satellite drop formation. Angular dependence of surface tension has been proposed numerically and validated for the respective case with the angle of 0° to deviate the least from literature value. The results obtained in this study are synchronous with the established theoretical understanding of the concept

Metabolic Evaluation in Pediatric Urolithiasis: Our Experience

Introduction: Pediatric urolithiasis is a morbid condition that is often associated with underlying metabolic disorders. It is essential to diagnose and manage the underlying metabolic disorder along with management of urolithiasis to prevent stone recurrence and decrease the morbidity.Materials and Methods: This retrospective observational study was conducted in 50 patients below 15 years of age with urolithiasis. Urine collection was done for 24 hours in toilet-trained children and spot samples were taken from younger children. The urinary parameters that were evaluated included calcium, oxalate, citrate, uric acid, and total urine volume. The serum levels of calcium, phosphorus, creatinine, uric acid, electrolytes, parathormone, and albumin were also measured. Stone analysis was done whenever possible.Results: There were 32 males and 18 females. Sixty-two percent of the patients had a low calcium intake and 70% of the children had a history of low water intake and had a low urine volume over 24 hours. Half of the children had serum metabolic abnormalities, including hypocalcaemia (n=19,38%), hypocalcaemia with hyperphosphatemia (n=2, 4%), hypercalcemia (n=2, 4%), and hyperuricemia (n=2,4%). Urinary abnormalities were detected in 42% of the children (n=21). These abnormalities included hypocitraturia in 11 patients (50%), hypercalciuria in 7 patients (30%), hyperoxaluria in 1 patient (6%), and hyperuricosuria in 2 patients (12%). Stone analysis was done in 18 patients. Fifteen patients (30%) had calcium oxalate stones, two patients (4%) had uric acid stones, and one patient had a mixed stone.Conclusions: It is important to maintain an optimal blood calcium level and increase fluid intake to prevent stone formation in children.Keywords: Urolithiasis; Pediatric; Metabolic Disorders

A machine learning approach for GRB detection in AstroSat CZTI data

We present a machine learning (ML) based method for automated detection of Gamma-Ray Burst (GRB) candidate events in the range 60 keV - 250 keV from the AstroSat Cadmium Zinc Telluride Imager data. We use density-based spatial clustering to detect excess power and carry out an unsupervised hierarchical clustering across all such events to identify the different light curves present in the data. This representation helps understand the instrument's sensitivity to the various GRB populations and identify the major non-astrophysical noise artefacts present in the data. We use Dynamic Time Warping (DTW) to carry out template matching, which ensures the morphological similarity of the detected events with known typical GRB light curves. DTW alleviates the need for a dense template repository often required in matched filtering like searches. The use of a similarity metric facilitates outlier detection suitable for capturing previously unmodelled events. We briefly discuss the characteristics of 35 long GRB candidates detected using the pipeline and show that with minor modifications such as adaptive binning, the method is also sensitive to short GRB events. Augmenting the existing data analysis pipeline with such ML capabilities alleviates the need for extensive manual inspection, enabling quicker response to alerts received from other observatories such as the gravitational-wave detectors

Rapalogs can promote cancer cell stemness in vitro in a Galectin-1 and H-ras-dependent manner

Currently several combination treatments of mTor- and Ras-pathway inhibitors are being tested in cancer therapy. While multiple feedback loops render these central signaling pathways robust, they complicate drug targeting. Here, we describe a novel H-ras specific feedback, which leads to an inadvertent rapalog induced activation of tumorigenicity in Ras transformed cells. We find that rapalogs specifically increase nanoscale clustering (nanoclustering) of oncogenic H-ras but not K-ras on the plasma membrane. This increases H-ras signaling output, promotes mammosphere numbers in a H-ras-dependent manner and tumor growth in ovo. Surprisingly, also other FKBP12 binders, but not mTor- inhibitors, robustly decrease FKBP12 levels after prolonged (> 2 days) exposure. This leads to an upregulation of the nanocluster scaffold galectin-1 (Gal-1), which is responsible for the rapamycin-induced increase in H-ras nanoclustering and signaling output. We provide evidence that Gal-1 promotes stemness features in tumorigenic cells. Therefore, it may be necessary to block inadvertent induction of stemness traits in H-ras transformed cells by specific Gal-1 inhibitors that abrogate its effect on H-ras nanocluster. On a more general level, our findings may add an important mechanistic explanation to the pleiotropic physiological effects that are observed with rapalogs.Peer reviewe