Two-Site Recognition of Phosphatidylinositol 3-Phosphate by PROPPINs in Autophagy

Macroautophagy is essential to cell survival during starvation and proceeds by the growth of a double-membraned phagophore, which engulfs cytosol and other substrates. The synthesis and recognition of the lipid phosphatidylinositol 3-phosphate, PI(3)P, is essential for autophagy. The key autophagic PI(3)P sensors, which are conserved from yeast to humans, belong to the PROPPIN family. Here we report the crystal structure of the yeast PROPPIN Hsv2. The structure consists of a seven-bladed β-propeller and, unexpectedly, contains two pseudo-equivalent PI(3)P binding sites on blades 5 and 6. These two sites both contribute to membrane binding in?vitro and are collectively required for full autophagic function in yeast. These sites function in concert with membrane binding by a hydrophobic loop in blade 6, explaining the specificity of the PROPPINs for membrane-bound PI(3)P. These observations thus provide a structural and mechanistic framework for one of the conserved central molecular recognition events in autophagy.     

Design of Site-Specific Microwave Phased Array Hyperthermia Applicators Using 434 MHz Reduced Cavity-Backed Patch Antenna

Cancers of the neck, breast, and lower extremities are common malignancies diagnosed in India with a higher incidence of advanced-stage disease. Phased array (PA) applicators reported for hyperthermia treatment (HT) of the breast have small focal region and high cross-coupling, and those reported for lower extremities provide regional heating and limited steering. In this study, we present the numerical design of site-specific PA applicators for HT of large solid tumors in the neck, breast, and lower extremities using a miniaturized 434 MHz cavity-backed water-loaded patch antenna. The fabricated antenna has 38 × 36 mm² aperture, more than 90% power coupling, 25 MHz bandwidth, and good agreement between simulated and measured specific absorption rate (SAR) in phantom. The site-specific applicators demonstrated less power reflection (<−17.9 dB) and cross-coupling (<−26.8 dB) for 5 mm inter-ring spacing. SAR indicators for 64 cc tumor at varying locations in simplified layered three-dimensional (3D) tissue models of the neck, breast, and leg showed average power absorption ratio (aPA_ratio) ≥ 3.16, target to hotspot quotient (THQ) ≥ 0.57, 25% iso-SAR coverage (TC₂₅) ≥ 81%, and 50% iso-SAR coverage (TC₅₀) ≥51.8%. Simulation results of site-specific applicators for 3D inhomogeneous patient models showed aPA_ratio ≥ 5.98, THQ ≥ 0.9, TC₅₀ ≥ 86%, and 100% TC₂₅ for all sites. It is concluded that the 434 MHz miniaturized cavity-backed patch antenna can be used to develop high-density PA applicators with 12–24 antennas for HT of large solid tumors (≥4 cm) in the neck, breast, and lower extremities with 3D steering ability and less cross-coupling (≤−26.8 dB). © 2020 Bioelectromagnetics Society     

Construction and characterization of a NotI-BsuE linking library from the human X chromosome.

We describe the construction and characterization of methylation-resistant sequence-tagged NotI linking clones specific for the X chromosome, referred to as NotI-BsuE linking clones. The approach consists of methylating the X-chromosome-specific cloned DNA with BsuE methylase (M. BsuE), an enzyme that methylates the first C residue in the CGCG sequence, followed by selection of the methylation-resistant NotI sites by insertion of a kanamycin-resistance gene in the clones cleavable by NotI. The frequent occurrence of NotI sites in CpG islands is expected to cause methylation of a large number of NotI sites with BsuE methylase, thereby rendering them resistant to NotI cleavage. Thus, the combination of M. BsuE and NotI yields less frequent cutting than the NotI alone. We have isolated, partially sequenced, and characterized 113 NotI-BsuE linking clones, and mapped 50 clones to various regions along the chromosome.     

In vitro propagation via organogenesis and embryogenesis of <Emphasis Type="Italic">Cyrtanthus mackenii</Emphasis>: a valuable threatened medicinal plant

Efficient and simple, organogenesis (direct and indirect) and somatic embryogenesis (cell suspension) systems were developed for in vitro propagation of Cyrtanthus mackenii, a valuable economic plant from leaf explants cultured on Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of sucrose, plant growth regulators (PGRs), glutamine, phloroglucinol (PG) and 6-(2-hydroxy-3-methylbenzylamino) purine (PI55). MS medium solidified with 8 g L^?1 agar (MS_S) containing 40 g L^?1 sucrose, 10 µM picloram, 2.5 µM benzyladenine (BA) and 20 µM glutamine produced a higher number of shoots from white nodular callus. This was however, not significantly different to direct shoot regeneration on media containing 10 µM picloram, 2.5 µM BA and a reduced concentration of sucrose and glutamine. The regenerated shoots were rooted best with MS_S medium incorporating 10 µM PG. The number of somatic embryos (SEs) were significantly higher using liquid MS medium containing 30 g L^?1 sucrose, 0.5 µM picloram, 1 µM thidiazuron or BA and 3 µM glutamine or gibberellic acid. The embryos were germinated in PGR-free MS_S medium. All plantlets were successfully acclimatized in the greenhouse. Histological studies confirmed the different developmental stages and bipolar structure of SE. The organogenesis and somatic embryogenesis protocols provides a system for large scale propagation and germplasm conservation. Developed protocols can be used for clonal production and pharmacological and genetic transformation studies.     

Biodegradation of gaseous toluene with mixed microbial consortium in a biofilter: steady state and transient operation

Bioprocess and Biosystems Engineering - Petroleum oil refineries are massive emitters of risky volatile organic compounds (VOCs). Among the VOCs, toluene is taken into account as a significant...     

Instability of CGG repeats in transgenic mice

Dynamic mutation resulting in the expansion of CGG repeats in the untranslated region (UTR) of the first exon of the FMR1 gene in humans results in fragile X syndrome. Long stretches of CGG repeats that are known to be highly unstable in humans have so far failed to show similar intergenerational instability in transgenic mice. We generated transgenic lines that show a dramatic increase from 26 to >300 repeats in three generations. One of the salient features of our transgene is the inclusion of the origin of replication of simian virus-40 (SV40), which is known to exclude nucleosomes. Three founder mice in FVB/NJ background show expansion of CGG repeats present in the transgene, supporting a postzygotic mechanism for CGG expansion that is independent of a genomic imprinting effect. We discuss here the results of analyzing one of the lines established.     

Therapeutic Potential of the Medicinal Plant <i>Tinospora cordifolia</i>–Minireview

For thousands of years, plant based herbal medicines have been utilized by millions of people all over the world. Plant materials or products are used in different folk/traditional medical systems, such as the Chinese, African and Indian medical systems, like Siddha, Ayurveda, Unani, and Homeopathy. Tinospora cordifolia (TC) is a medicinal plant belonging to the family Menispermaceae. It is a big deciduous, climbing shrub growing prevalently in the tropical part of Indian subcontinent regions such as India, Pakistan, Nepal, Bhutan, Bangladesh and Srilanka, and in Myanmar, and China. Guduchi, Giloy, Shindilkodi, and Amritha are all the common names for this plant. Extracts from different parts of this herbal plant have been used to treat many diseases. In Ayurvedic medicine, extract from this plant is used for preparing “rasayanas”, which is known to cure diabetes, skin diseases, allergic conditions, jaundice, cardiovascular diseases, rheumatoid arthritis, poisoning, and microbial infections. T. cordifolia has a many bioactive phytochemicals that have been isolated from its aerial parts and roots. Many bioactive principles have been reported from this plant which belong to various classes like alkaloids, aliphatic compounds, diterpenoid lactones, phenolics, flavonoids, glycosides, sesquiterpenoids, lignans, steroids and polysaccharides. T. cordifolia possesses medicinal properties such as antioxidant, antiallergic, antiinflammatory, antimicrobial, antiviral, antidote, antitumor, antileprotic, antispasmodic, and antidiabetic properties. The present review will provide a comprehensive therapeutic potential of T. cordifolia.     

Ectomycorrhiza,Friend or Foe?

Ecosystems - Many ecology textbooks present the interaction between mycorrhizal fungi and their host plants as the archetype of symbiosis or mutualism. However, mycorrhiza drains carbon directly...     

Synthesis, cytotoxicity, and antiviral activities of new neolignans related to honokiol and magnolol

A series of new bisphenol derivatives bearing allylic moieties were synthesized as potential analogs of honokiol and/or magnolol. Certain compounds exhibited specific anti-proliferation activity against SVR cells and moderate anti-HIV-1 activity in primary human lymphocytes. Compound 5h was the most potent compound and its anti-tumor activity was evaluated in vivo.