Molecular farming for sustainable production of clinical-grade antimicrobial peptides.

Antimicrobial peptides (AMPs) are emerging as next-generation therapeutics due to their broad-spectrum activity against drug-resistant bacterial strains and their ability to eradicate biofilms, modulate immune responses, exert anti-inflammatory effects and improve disease management. They are produced through solid-phase peptide synthesis or in bacterial or yeast cells. Molecular farming, i.e. the production of biologics in plants, offers a low-cost, non-toxic, scalable and simple alternative platform to produce AMPs at a sustainable cost. In this review, we discuss the advantages of molecular farming for producing clinical-grade AMPs, advances in expression and purification systems and the cost advantage for industrial-scale production. We further review how 'green' production is filling the sustainability gap, streamlining patent and regulatory approvals and enabling successful clinical translations that demonstrate the future potential of AMPs produced by molecular farming. Finally, we discuss the regulatory challenges that need to be addressed to fully realize the potential of molecular farming-based AMP production for therapeutics.

Efficient in planta production of amidated antimicrobial peptides that are active against drug-resistant ESKAPE pathogens.

Antimicrobial peptides (AMPs) are promising next-generation antibiotics that can be used to combat drug-resistant pathogens. However, the high cost involved in AMP synthesis and their short plasma half-life render their clinical translation a challenge. To address these shortcomings, we report efficient production of bioactive amidated AMPs by transient expression of glycine-extended AMPs in Nicotiana benthamiana line expressing the mammalian enzyme peptidylglycine α-amidating mono-oxygenase (PAM). Cationic AMPs accumulate to substantial levels in PAM transgenic plants compare to nontransgenic N. benthamiana. Moreover, AMPs purified from plants exhibit robust killing activity against six highly virulent and antibiotic resistant ESKAPE pathogens, prevent their biofilm formation, analogous to their synthetic counterparts and synergize with antibiotics. We also perform a base case techno-economic analysis of our platform, demonstrating the potential economic advantages and scalability for industrial use. Taken together, our experimental data and techno-economic analysis demonstrate the potential use of plant chassis for large-scale production of clinical-grade AMPs.

A pro-oxidant combination of resveratrol and copper down-regulates multiple biological hallmarks of ageing and neurodegeneration in mice.

Billions of cells die in the body every day, and cell-free chromatin particles (cfChPs) which are released from them enter into the extracellular compartments of the body, including into the circulation. cfChPs are known to readily enter into healthy cells to damage their DNA and activate apoptotic and inflammatory pathways. We have hypothesized that lifelong assault on healthy cells by cfChPs is the underlying cause of ageing, and that ageing could be retarded by deactivating extra-cellular cfChPs. The latter can be effected by oxygen radicals that are generated upon admixing the nutraceuticals resveratrol and copper (R-Cu). The present study investigated whether prolonged administration of R-Cu would retard biological hallmarks of ageing. C57Bl/6 mice were divided into 3 equal groups; one group was sacrificed at age 3 months, and which acted as young controls. The remaining mice were allowed to age, and at age 10 months the experimental ageing group was given R-Cu by oral gavage twice daily for further 12 months at a dose of 1 mg/kg of R and 0.1 µg/kg of Cu. The control ageing group was given water by oral gavage twice daily for 12 months. Animals of both groups were sacrificed at age 22 months. R-Cu treatment led to reduction of several biological hallmarks of ageing in brain cells which included telomere attrition, amyloid deposition, DNA damage, apoptosis, inflammation, senescence, aneuploidy and mitochondrial dysfunction. R-Cu treatment also led to significant reduction in blood levels of glucose, cholesterol and C-reactive protein. These findings suggest that cfChPs may act as global instigators of ageing and neurodegeneration, and that therapeutic use of R-Cu may help to make healthy ageing an attainable goal.

Impact of Thermal Fluctuations on Logarithmic Corrected Massive Gravity Charged Black Hole.

We investigate the influence of the first-order correction of entropy caused by thermal quantum fluctuations on the thermodynamics of a logarithmic corrected charged black hole in massive gravity. For this black hole, we explore the thermodynamic quantities, such as entropy, Helmholtz free energy, internal energy, enthalpy, Gibbs free energy and specific heat. We discuss the influence of the topology of the event horizon, dimensions and nonlinearity parameter on the local and global stability of the black hole. As a result, it is found that the holographic dual parameter vanishes. This means that the thermal corrections have no significant role to disturb the holographic duality of the logarithmic charged black hole in massive gravity, although the thermal corrections have a substantial impact on the thermodynamic quantities in the high-energy limit and the stability conditions of black holes.

Cell-free chromatin: A newly described mediator of systemic inflammation.

Recent research has shown that cell-free chromatin (cfCh) particles that are released from the billions of cells that die in the body everyday can enter into healthy cells, integrate into their genomes and induce dsDNA breaks and apoptotic responses. Genomic integration of cfCh activates NF κ B suggesting a novel mechanism of induction of systemic inflammation. Since DNA damage and inflammation are underlying pathologies in multiple devastating acute and chronic disease conditions, the discovery of agents that can inactivate cfCh may provide therapeutic possibilities.

Illegitimate and Repeated Genomic Integration of Cell-Free Chromatin in the Aetiology of Somatic Mosaicism, Ageing, Chronic Diseases and Cancer.

Emerging evidence suggests that an individual is a complex mosaic of genetically divergent cells. Post-zygotic genomes of the same individual can differ from one another in the form of single nucleotide variations, copy number variations, insertions, deletions, inversions, translocations, other structural and chromosomal variations and footprints of transposable elements. High-throughput sequencing has led to increasing detection of mosaicism in healthy individuals which is related to ageing, neuro-degenerative disorders, diabetes mellitus, cardiovascular diseases and cancer. These age-related disorders are also known to be associated with significant increase in DNA damage and inflammation. Herein, we discuss a newly described phenomenon wherein the genome is under constant assault by illegitimate integration of cell-free chromatin (cfCh) particles that are released from the billions of cells that die in the body every day. We propose that such repeated genomic integration of cfCh followed by dsDNA breaks and repair by non-homologous-end-joining as well as physical damage to chromosomes occurring throughout life may lead to somatic/chromosomal mosaicism which would increase with age. We also discuss the recent finding that genomic integration of cfCh and the accompanying DNA damage is associated with marked activation of inflammatory cytokines. Thus, the triple pathologies of somatic mosaicism, DNA/chromosomal damage and inflammation brought about by a common mechanism of genomic integration of cfCh may help to provide an unifying model for the understanding of aetiologies of the inter-related conditions of ageing, degenerative disorders and cancer.

Is inflammation a direct response to dsDNA breaks?

Recent research shows that extra-nuclear cell-free chromatin (cfCh) released from dying cells can freely enter into healthy cells and integrate into their genomes. Genomic integration of cfCh leads to dsDNA breaks and activation of inflammatory cytokines both of which occur concurrently with similar kinetics and that induction of inflammation can be abrogated by preventing DNA breaks with the use of cfCh inactivating agents. The proposal is put forward that inflammatory cytokines are a new family of DDR proteins that are activated following dsDNA breaks inflicted by genomic integration of cfCh.

Current challenges in the therapeutic use of induced pluripotent stem cells (iPSCs) in cancer therapy

Induced Pluripotent Stem Cells (iPSCs) technology has catapulted the field of stem-cell biology through ectopic expression of reprogramming factors. Ever since its discovery, the potential of iPSCs has been explored by many scientists to unravel the molecular mechanism responsible for cancer initiation and progression. Besides modeling cancer, the further applications of this technology includes high-throughput drug screening, epigenetic reprogramming of cancer cell state to normal, immunotherapy and regenerative cell therapies. Here, we review the current challenges on clinical applications of iPSCs with respect to understanding cancer and personalizing treatment for the disease (AU)

Preliminary physicochemical evaluation of Kushta tutia: A Unani Formulation.

BACKGROUND: Kushta is an important solid dosage form of Unani system of medicine used to treat various ailments. Very small particle size of kushta is responsible for its rapid absorption in body leading to instant therapeutic actions. Kushta tutia (KT) is one such renowned formulation used by hakims for successful management of various disorders. However, there is lack of scientific work on KT. OBJECTIVES: The present study was performed to evaluate KT physicochemically by testifying it on classical tests along with modern scientific techniques. MATERIALS AND METHODS: Tutia was first detoxified as per classical literature. It was triturated with water and dried, afterwards subjected to calcination in furnace rather than cow dung cakes due to isolation of material being heated and better temperature control. Finished product was evaluated for physicochemical characteristics including preliminary tests mentioned in classical literature. RESULTS: Floating and finger test were positive. Curd test showed no discoloration after 48 h. These findings indicate correct preparation of KT according to classical literature. Bulk density (0.96 ± 0.00 g/ml); tapped density (1.53 ± 0.00 g/ml); Hausner ratio (0.62 ± 0.00), compressibility index (37.52 ± 0.19%); loss of weight on drying (0.08 ± 0.00%); pH of 1 and 10% (5.20 ± 0.00) and 5.62 ± 0.00, respectively); total ash, acid insoluble ash, and water soluble ash values 95.75 ± 0.09, 6.57 ± 0.02, and 45.02 ± 0.20%, respectively; and extractive values 0.85 ± 0.02% were reported in KT. CONCLUSION: Since this work has not been reported earlier, the results obtained could be considered as the standard for KT for future studies.

The In vitro anti-acne activity of two unani drugs.

BACKGROUND: Acne is the most common disorder treated by dermatologists. As many as 80-90% of all adolescents have some type of acne and 30% of them require medical treatment. It is an inflammatory disease of the pilosebaceous unit characterized by the formation of open and closed comedones, papules, pustules, nodules, and cysts. AIMS: The present study was conducted to investigate the in vitro anti-acne activity of two Unani single drugs Darchini (Cinnamomum zeylanicum Bl.) and Tukhm Khashkhash (Papaver somniferum L. seeds). MATERIALS AND METHODS: The antibacterial activity of aqueous, ethanolic and hydroalcoholic extracts of both drugs were investigated against two acne causing bacteria, i.e., Propionibacterium acne and Staphylococcus epidermidis using well diffusion method. RESULTS: The result showed that both drugs were active against the two bacteria. Against P. acne aqueous and ethanolic extract of Darchini and Tukhm Khashkhash showed the zone of inhibition of 18 ± 1.02 mm and 18 ± 1.6 mm and 13 ± 1.04 mm and 14 ± 1.8 mm, respectively. Against S. epidermidis aqueous, hydroalcoholic and ethanolic extracts of Darchini showed 22 ± 1.7 mm, 22 ± 1.2 mm and 15 ± 1.8 mm zone of inhibition respectively. Hydroalcoholic and ethanolic extracts of Tukhm Khashkhash showed 15 ± 1.09 mm and 13 ± 1.6 mm zone of inhibition respectively. CONCLUSION: This suggests that C. zeylanicum and P. somniferum have potential against acne causing bacteria and hence they can be used in topical anti-acne preparations and may address the antibiotic resistance of the bacteria.