The roles of initiation factor 2 and guanosine triphosphate in initiation of protein synthesis

The role of IF2 from Escherichia coli was studied in vitro using a system for protein synthesis with purified components. Stopped flow experiments with light scattering show that IF2 in complex with guanosine triphosphate (GTP) or a non-cleavable GTP analogue (GDPNP), but not with guanosine diphosphate (GDP), promotes fast association of ribosomal subunits during initiation. Biochemical experiments show that IF2 promotes fast formation of the first peptide bond in the presence of GTP, but not GDPNP or GDP, and that IF2-GDPNP binds strongly to post-initiation ribosomes. We conclude that the GTP form of IF2 accelerates formation of the 70S ribosome from subunits and that GTP hydrolysis accelerates release of IF2 from the 70S ribosome. The results of a recent report, suggesting that GTP and GDP promote initiation equally fast, have been addressed. Our data, indicating that eIF5B and IF2 have similar functions, are used to rationalize the phenotypes of GTPase-deficient mutants of eIF5B and IF2.     

Crude alkaloid extract of Rhazya stricta inhibits cell growth and sensitizes human lung cancer cells to cisplatin through induction of apoptosis

There is an urgent need to improve the clinical management of non-small cell lung cancer (NSCLC), one of the most frequent causes of cancer-related deaths in men and women worldwide. Rhazya stricta, an important medicinal plant used in traditional Oriental medicine, possesses anti-oxidant, anti-carcinogenic and free radical scavenging properties. This study was done to explore the potential anticancer activity of a crude alkaloid extract of R. stricta (CAERS) against the NSCLC line A549. CAERS markedly suppressed the growth of A549 cells and considerably enhanced the anti-proliferative potential of cisplatin. CAERS-mediated inhibition of A549 cell growth correlated with the induction of apoptosis that was accompanied by numerous morphological changes, DNA fragmentation, an increase in the Bax/Bcl-2 ratio, the release of mitochondrial cytochrome c, activation of caspases 3 and 9 and cleavage of poly(ADP-ribose)-polymerase. CAERS reduced the constitutive expression of anti-apoptotic proteins (Bcl-2, Bcl-X_L, Mcl-1 and Survivin) and cell cycle regulating proteins (cyclin D1 and c-Myc), but enhanced expression of the proapoptotic proteins Noxa and BAD. These observations indicate that CAERS induced apoptosis and sensitized NSCLC to cisplatin via a mitochondria-mediated apoptotic pathway. These data provide a rationale for using a combination of CAERS and CDDP to treat NSCLC and other CDDP-resistant tumors.     

Potentiality of Different Seed Priming Agents to Mitigate Cold Stress of Winter Rice Seedling

Seed priming has proved to be an effective pre-germination seed invigoration technique for different crops to improve seed and seedling performance under different abiotic stresses. In Bangladesh, winter rice is very often exposed to cold waves just after sowing in the nursery bed resulting in poor seed germination and seedling emergence, yellowish and thin seedlings production, and a very low survival rate. Seed priming may mitigate the cold stress during seed germination and seedling emergence and helps in the quality seedling production of winter rice. To evaluate the efficacy of different seed priming techniques in increasing seedling emergence, growth, vigor and survivability of winter rice cultivars under cold stress, a pot experiment was conducted at the Department of Agronomy, Bangladesh Agricultural University during December 2018 to January 2019. The experiment comprised two factors, (A) Winter rice variety namely, i) BRRI dhan29 and ii) BRRI dhan36; (B) Seed priming agent namely i) Control (no priming), ii) 20000 ppm NaCl, iii) 30000 ppm NaCl, iv) 20000 ppm KCl, v) 30000 ppm KCl, vi) 20000 ppm CaCl₂, vii) 30000 ppm CaCl₂, viii) 50 ppm CuSO₄, ix) 75 ppm CuSO₄, x) 10000 ppm ZnSO₄, xi) 15000 ppm ZnSO₄, xii) 2 ppm Na₂MoO₄, xiii) 3 ppm Na₂MoO₄, xiv) 100 ppm PEG (Polyethylene glycol 4000) and xv) 150 ppm PEG. Seeds were sown on two different dates viz., 1st December and 1st January so that seedlings are exposed to cold stress at different stages. The experiment was laid out in a completely randomized design (CRD) with three replications. Results indicated that (in most of the cases) seed priming has a positive impact on seedling emergence rate (%), root length, shoot length, root shoot ratio, root dry weight, shoot dry weight, seedling dry weight and survival rate (%). Among the priming agents, KCl and CaCl₂ performed best; while priming with NaCl and PEG showed no advantages over no priming for both the sowing dates. In general, BRRI dhan36 performed better than BRRI dhan29 in terms of seedling growth because of its higher tolerance to cold stress. But, both the varieties performed similarly in terms of emergence rate and survival rate. Thus, priming is an effective tool to increase seed germination, better seedling growth, and higher seedling survivability of winter rice under cold stress, and KCl (20000 ppm) or CaCl₂ (20000 ppm) can be considered as a viable priming agent.     

Rebound of hepatitis B virus replication in HepG2 cells after cessation of antiviral treatment

Treatment of patients with lamivudine (3TC) results in loss of detectable levels of hepatitis B virus (HBV) DNA from serum; however, the relapse rate, with regard to both reappearance of virus in the bloodstream and hepatic inflammation, is high when therapy is terminated. Although the rebound observed in patients has also been seen in animal hepadnavirus models, rebound has not been analyzed in an in vitro cell culture system. In this study, we used the HBV recombinant baculovirus/HepG2 system to measure the time course of antiviral agent-mediated loss of HBV replication as well as the time course and magnitude of HBV production after release from antiviral treatment. Because of the sensitivity of the system, it was possible to measure secreted virions, intracellular replicative intermediates, and nuclear non-protein-bound HBV DNA and separately analyze individual species of DNA, such as single-stranded HBV DNA compared to the double-stranded form and relaxed circular compared to covalently closed circular HBV DNA. We first determined that HBV replication in the HBV recombinant baculovirus/HepG2 system could proceed for at least 35 days, with a 30-day plateau level of replication, making it possible to study antiviral agent-mediated loss of HBV followed by rebound after cessation of drug treatment. All HBV DNA species decreased in a time-dependent fashion following antiviral treatment, but the magnitude of decline differed for each HBV DNA species, with the covalently closed circular form of HBV DNA being the most resistant to drug therapy. When drug treatment ceased, HBV DNA species reappeared with a pattern that recapitulated the initiation of replication, but with a different time course.     

Partial purification and characterization of 2, 4-diacetylphloroglucinol producing Pseudomonas fluorescens VSMKU3054 against bacterial wilt disease of tomato

We find out the antimicrobial potential of partially purified 2,4-diacetylphloroglucinol (DAPG) against Ralstonia solanacearum and fungal plant pathogens isolated from tomato rhizobacterium Pseudomonas fluorescens VSMKU3054. The present study is mainly focused on the control of wilt disease of tomato by our isolate VSMKU3054 and DAPG. The cell free culture filtrate of P. fluorescens VSMKU3054 was significantly arrested the growth of R. solanacearum and fungal pathogens such as Rhizoctonia solani, Sclerotium rolfsii, Macrophomina phaseolina and Fusarium oxysporum compared to control. The existence of DAPG from the crude metabolites of P. fluorescens VSMKU3054 was confirmed on TLC with Rf value 0.34, which is coincide with that of authentic phloroglucinol. The partially purified DAPG exhibited much higher activity against R. solanacearum at 30 µg/ml than the fungal plant pathogens compared to control. The antimicrobial partially purified compound was identified as DAPG by UV, FT-IR and GC–MS analysis. The percentage of live cells of R. solanacearum when supplemented with DAPG at 30 µg/ml, significantly controlled the living nature of R. solanacearum up to 68% compared to tetracycline and universal control observed under high content screening analysis. The selected isolate P. fluorescens VSMKU3054 and DAPG significantly controlled wilt disease of tomato up to 59.5% and 42.12% on 3rd and 7th days compared to positive and negative control by detached leaf assay. Further, in silico analysis revealed that high interaction of DAPG encoding protease with lectin which is associated with R. solanacearum. Based on our findings, we confirmed that P. fluorescens VSMKU3054 and DAPG could be used a potential bio inoculants for the management of bacterial wilt disease of tomato.     

Stability and anatomical parameters of irradiated potato cultivars under drought stress

This study was carried out in Desert Research Center and Faculty of Agriculture, Zagazig University, Egypt, under North Sinai conditions during three growing seasons, i.e., summer 2018, fall 2018/2019 and summer 2019 to assess the effect of radiation mutants on leaf histological features and genetic stability of the productivity of some potato cultivars under drought stress conditions. Results reveal that the genotypes can be statistically classified based on regression coefficient (bi), deviation from regression (S²di) to 4 groups (with low in S²di are considered in all groups) as: (i) Genotype with elevated average, bi = 1, it is considered as stable genotype where Cara cultivar (both generations) was included. (ii) Genotype with elevated average, bi > 1 as genotype with average stability where spunta cultivar was involved. (iii) Genotype with low mean, bi < 1 as genotype with low stability where hermes (both primary (M₁) and secondary (M₂) radiated generations) and Caruso (2nd generation) cultivars were involved. (iv) Genotypes including a few bi values in one generation, as genotype including low stability but are not recommended for use in this generation where Caruso cv in M₁ was included. The results indicated that 20 Gy irradiation exposure revealed that Spunta cultivar produced markedly high mean combined over yield during M₁ (11.771 ton/fed) and M₂ (10.97 ton/fed) generations than other genotypes and ranked first over all environments. It proves that Hermes could be employed as anti-stress genotypes under stress conditions (negative conditions or poor yielding). However, spunta followed by cara cv. represented a good performance in M₂ production yield (10.97 and 8.51 ton/fed, respectively), slight drift from the regression line and coefficient close to 1, therefore, both cultivars were excellent between genotypes in shape of yield stability and is recommended for different conditions. According to anatomical studies, 80 % from field capacity (FC) decreased the thickness of medvein and lamina of potato cv. spunta, also, dimensions of medvein bundle and mean diameter of vessels. In conclusion, plants treated with gamma ray at level 20 and grown under 80 % FC induced prominent increase in all previous characters.     

Signaling Pathways in Leiomyoma: Understanding Pathobiology and Implications for Therapy

Uterine leiomyomas are the most common tumors of the female genital tract, affecting 50% to 70% of females by the age of 50. Despite their prevalence and enormous medical and economic impact, no effective medical treatment is currently available. This is, in part, due to the poor understanding of their underlying pathobiology. Although they are thought to start as a clonal proliferation of a single myometrial smooth muscle cell, these early cytogenetic alterations are considered insufficient for tumor development and additional complex signaling pathway alterations are crucial. These include steroids, growth factors, transforming growth factor-beta (TGF-β)/Smad; wingless-type (Wnt)/β-catenin, retinoic acid, vitamin D, and peroxisome proliferator-activated receptor γ (PPARγ). An important finding is that several of these pathways converge in a summative way. For example, mitogen-activated protein kinase (MAPK) and Akt pathways seem to act as signal integrators, incorporating input from several signaling pathways, including growth factors, estrogen and vitamin D. This underlines the multifactorial origin and complex nature of these tumors. In this review, we aim to dissect these pathways and discuss their interconnections, aberrations and role in leiomyoma pathobiology. We also aim to identify potential targets for development of novel therapeutics.