Cloning of hok gene into anhydrotetracycline inducible pASK75 vector reveals potent antimicrobial effect of 19 amino acid long N-terminal fragment of hok peptide

An important toxin-antitoxin (TA) system hok/sok, encoded by R1 plasmid of Escherichia coli, is involved in the post segregation killing of cells that have lost the plasmid. The lethal properties of hok protein have been utilized for the environmental containment of microbes and the development of potential vaccine candidates. This study aimed to demonstrate the potent anti-microbial property of a 19 amino acid (AA) long N-terminal fragment of hok peptide. This was accomplished by designing a conditional suicide system based on hok gene expression cloned in an anhydrotetracycline (aTc) inducible vector – pASK75. Heat shock and electroporation were utilized for the transformation of Escherichia coli and Vibrio cholerae cells, respectively. The minimal induction concentration (MI_dC) of aTc, determined by analyzing the expression of green fluorescent protein cloned separately into pASK75 vector, was 30 ng/mL. As hok gene was synthesized de novo (using recombinant polymerase chain reaction) in our study, various random sized hok fragments were generated (as a result of the error-prone nature of Taq polymerase). The smallest hok fragment able to bring about effective antimicrobial killing was a 19 AA long N-terminal fragment of hok having the wild type sequence, except for the carboxy terminus AA residue. The MI_dC of aTc in our experiments was 6-fold lower than previously reported, making our bacterial clones suitable for use in mammalian systems as potential vaccine candidates. Based on our experiments, we hypothesize the 19 AA long N-terminal fragment of hok peptide to be the smallest possible hok fragment sufficient to bring about effective antimicrobial killing.     

The bone microenvironment increases phenotypic plasticity of ER+ breast cancer cells

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Advances in Metered Dose Inhaler Technology: Hardware Development

Pressurized metered dose inhalers (MDIs) were first introduced in the 1950s and they are currently widely prescribed as portable systems to treat pulmonary conditions. MDIs consist of a formulation containing dissolved or suspended drug and hardware needed to contain the formulation and enable efficient and consistent dose delivery to the patient. The device hardware includes a canister that is appropriately sized to contain sufficient formulation for the required number of doses, a metering valve capable of delivering a consistent amount of drug with each dose delivered, an actuator mouthpiece that atomizes the formulation and serves as a conduit to deliver the aerosol to the patient, and often an indicating mechanism that provides information to the patient on the number of doses remaining. This review focuses on the current state-of-the-art of MDI hardware and includes discussion of enhancements made to the device’s core subsystems. In addition, technologies that aid the correct use of MDIs will be discussed. These include spacers, valved holding chambers, and breath-actuated devices. Many of the improvements discussed in this article increase the ability of MDI systems to meet regulatory specifications. Innovations that enhance the functionality of MDIs continue to be balanced by the fact that a key advantage of MDI systems is their low cost per dose. The expansion of the health care market in developing countries and the increased focus on health care costs in many developed countries will ensure that MDIs remain a cost-effective crucial delivery system for treating pulmonary conditions for many years to come.     

Castasterone and Citric Acid Supplementation Alleviates Cadmium Toxicity by Modifying Antioxidants and Organic Acids in Brassica juncea

Journal of Plant Growth Regulation - Brassinosteroids are known to manage various stresses including heavy metals in plants and castasterone is a bioactive form of brassinosteroids. Citric acid, a...     

Research progress in allele-specific expression and its regulatory mechanisms

Although the majority of genes are expressed equally from both alleles, some genes are differentially expressed. Organisms possess characteristics to preferentially express a particular allele under regulatory factors, which is termed allele-specific expression (ASE). It is one of the important genetic factors that lead to phenotypic variation and can be used to identify the variance of gene regulation factors. ASE indicates mechanisms such as DNA methylation, histone modifications, and non-coding RNAs function. Here, we review a broad survey of progress in ASE studies, and what this simple yet very effective approach can offer in functional genomics, and possible implications toward our better understanding of the underlying mechanisms of complex traits.     

The Influence of Environmental Conditions on Secondary Metabolites in Medicinal Plants: A Literature Review

Medicinal plants, a source of different phytochemical compounds, are now subjected to a variety of environmental stresses during their growth and development. Different ecologically limiting factors including temperature, carbon dioxide, lighting, ozone, soil water, soil salinity and soil fertility has significant impact on medicinal plants′ physiological and biochemical responses, as well as the secondary metabolic process. Secondary metabolites (SMs) are useful for assessing the quality of therapeutic ingredients and nowadays, these are used as important natural derived drugs such as immune suppressant, antibiotics, anti-diabetic, and anti-cancer. Plants have the ability to synthesize a variety of secondary metabolites to cope with the negative effects of stress. Here, we focus on how individual environmental variables influence the accumulation of plant secondary metabolites. A total of 48 articles were found to be relevant to the review topic during our systematic review. The review showed the influence of different environmental variables on SMs production and accumulation is complex suggesting the relationship are not only species-specific but also related to increases and decline in SMs by up to 50 %. Therefore, this review improves our understanding of plant SMs ability to adapt to key environmental factors. This can aid in the efficient and long-term optimization of cultivation techniques under ambient environmental conditions in order to maximize the quality and quantity of SMs in plants.     

Advances in Metered Dose Inhaler Technology: Formulation Development

Pressurized metered dose inhalers (MDIs) are a long-standing method to treat diseases of the lung, such as asthma and chronic obstructive pulmonary disease. MDIs rely on the driving force of the propellant, which comprises the bulk of the MDI formulation, to atomize droplets containing drug and excipients, which ideally should deposit in the lungs. During the phase out of chlorofluorocarbon propellants and the introduction of more environmentally friendly hydrofluoroalkane propellants, many improvements were made to the methods of formulating for MDI drug delivery along with a greater understanding of formulation variables on product performance. This review presents a survey of challenges associated with formulating MDIs as solution or suspension products with one or more drugs, while considering the physicochemical properties of various excipients and how the addition of these excipients may impact overall product performance of the MDI. Propellants, volatile and nonvolatile cosolvents, surfactants, polymers, suspension stabilizers, and bulking agents are among the variety of excipients discussed in this review article. Furthermore, other formulation approaches, such as engineered excipient and drug-excipient particles, to deliver multiple drugs from a single MDI are also evaluated.     

Cadmium Alters Mitochondrial Membrane Potential,Inhibits Electron Transport Chain Activity and Induces Callose Deposition in Rice Seedlings

Journal of Plant Growth Regulation - In the present study, seedlings of indica rice (Oryza sativa L.) cultivars were subjected to 100 and 200 µM Cd(NO3)2 treatment in hydroponics for...     

Shift in Cyanobacteria Community Diversity in Hot Springs of India

Two ecologically distinct tropical sulfur-rich alkaline hot springs, Taptapani at 48°C harboring mesophiles and Atri at 58°C comprising thermophiles situated in the Eastern Ghats foothills of India, differ in their geochemical conditions, and provide an interesting platform to unravel the eco-physiological reasons behind the differential cyanobacterial diversity. The predominance of mesophilic Arthronema (83.81%) in Taptapani and shifting predominance of thermophilic Leptolyngbya (96.25%) in Atri as discovered through 16S rRNA gene Illumina sequencing of their metagenomics DNA as a function of temperature are the intriguing features of the present study. Differential presence of the cyanobacterial community at the phylum level in these two hot springs was found to be correlated with the unequal coexistence of Chloroflexi, Taptapani the non-cyanobacteria members and the possible influence of physiochemical parameters including temperature. Variation in cyanobacterial diversity and composition of these hot springs as revealed through sequence analysis were also evinced by respective differences in richness, evenness, and Shannon diversity indices.