Direct Nuclear Delivery of Proteins on Living Plant via Partial Enzymatic Cell Wall Digestion.

Nuclear protein delivery underlies an array of biotechnological and therapeutic applications. While many variations of protein delivery methods have been described, it can still be difficult or inefficient to introduce exogenous proteins into plants. A major barrier to progress is the cell wall which is primarily composed of polysaccharides and thus only permeable to small molecules.

Structure, Absolute Configuration, Antiproliferative and Phytotoxic Activities of Icetexane and Abietane Diterpenoids from and Chemotaxonomic Implications.

From the aerial parts of Zamudio and Bedolla, three new icetexane-type diterpenoids were isolated. Their structures were established through spectroscopic methods and named the following: salvicarranzanolide (), 19-deoxo-salvicarranzanolide () and 19-deoxo-20-deoxy-salvicarranzanolide (). In addition, the known icetexane-type diterpenoids, 6,7,11,14-tetrahydro-7-oxo-icetexone (), -icetexone (), 19-deoxo--icetexone (), icetexone (), 19-deoxo-icetexone () and 7α-acetoxy-6,7-dihydroicetexone (), were also isolated, along with the abietanes sessein () and ferruginol ().

detected in the oral cavity of a dog in Kansas.

is an emerging human fungal pathogen, first described in Japan in 2009, and first detected in the United States in 2016. Here, we report the first-ever description of colonizing a human pet, the first identification of in a non-human mammal in the United States and the first isolate from the state of Kansas. While analyzing the oral mycobiome of dogs from a shelter in Kansas, the oral swab from one dog was found to contain as well as three other fungal species.

Fungal diversity and drug susceptibility of the oral mycobiome of domestic dogs.

The purpose of this study was to characterize the variety and diversity of the oral mycobiome of domestic dogs and to identify the commensal and potentially pathogenic fungi present. Two hundred fifty-one buccal swabs from domestic dogs were obtained and struck onto a chromogenic fungal growth medium that distinguishes between fungal species based on colony color and morphology. After isolating and harvesting single colonies, genomic DNA was extracted from pure cultures.

Transport Across Membranes: Techniques for Measuring Drug Import in Fungal Cells.

The ability of many antifungal molecules to traverse the fungal cell wall and accumulate within the cell is crucial to its ability to have the desired biological activity. Altered accumulation of the drug is an important mechanism of antifungal drug resistance. The best characterized mechanism for altered accumulation is through the action of the drug efflux pump which actively transports the drugs out of the membrane, although this is not the only mechanism for this phenomenon.

Transport across Membranes: Techniques for Measuring Efflux in Fungal Cells.

One of the most prevalent mechanisms of antifungal drug resistance is export of the molecule from the fungal cells through the action of putative efflux pumps or transporters. Drug efflux is a particularly common mechanism of resistance to azole antifungals, one of the most widely used classes of antifungal drugs. Here, we provide detailed protocols for two assays of small-molecule efflux activity: rhodamine 6G efflux and alanine-naphthylamide accumulation.

Inositol Phosphoryl Transferase, Ipt1, Is a Critical Determinant of Azole Resistance and Virulence Phenotypes in .

In this study, we have specifically blocked a key step of sphingolipid (SL) biosynthesis in by disruption of the orthologs of ScIpt1 and ScSkn1. Based on their close homology with counterparts, the proteins are predicted to catalyze the addition of a phosphorylinositol group onto mannosyl inositolphosphoryl ceramide (MIPC) to form mannosyl diinositolphosphoryl ceramide (M(IP)C), which accounts for the majority of complex SL structures in membranes. High throughput lipidome analysis confirmed the accumulation of MIPC structures in and cells, albeit to lesser extent in the latter.

Unmasking of CgYor1-Dependent Azole Resistance Mediated by Target of Rapamycin (TOR) and Calcineurin Signaling in Candida glabrata.

In this study, 18 predicted membrane-localized ABC transporters of Candida glabrata were deleted individually to create a minilibrary of knockouts (KO). The transporter KOs were analyzed for their susceptibility toward antimycotic drugs. Although Cg has previously been reported to be upregulated in various azole-resistant clinical isolates of C.

Application of the community dialogues method to identify ethical values and priorities related to pharmacogenomics.

Given the expansion of genetics in medicine, there is a growing need to develop approaches to engage patients in understanding how genetics affects their health. Various qualitative methods have been applied to gain a deeper understanding of patient perspectives in topics related to genetics. Community dialogues (CD) are a bi-directional research method that invites community members to discuss a pertinent, challenging topic over the course of a multi-week period and the community members openly discuss their positions on the topic.

Implementation of a Pharmacogenomic Testing Service through Community Pharmacy in the Netherlands: Results from an Early Service Evaluation.

Community pharmacy services have evolved to include medical and pharmaceutical interventions alongside dispensing. While established pharmacogenomic (PGx) testing is available throughout the Netherlands, this is primarily based in hospital environments and for specialist medicines. The aim of this work was to describe how best to implement PGx services within community pharmacy, considering potential barriers and enablers to service delivery and how to address them.

Structural Elucidation of Malonylcommunol and 6β-Hydroxy--communic Acid, Two Undescribed Diterpenes from . First Examples of Labdane Diterpenoids from a Mexican Species.

The aerial parts of afforded two undescribed labdane diterpenoids and (malonylcommunol and 6β-hydroxy--communic acid) along with two known labdane diterpenoids, -communic acid () and -communol (). Additionally, seven known metabolites were also isolated; two isopimarane diterpenoids and , two sesquiterpenoids identified as β-eudesmol () and cryptomeridiol (), and three aromatic compounds identified as phthalic acid (), a mixture of tyrosol fatty acid esters () and the flavone salvigenine (). While compounds compounds - showed significant inhibition of yeast α-glucosidase, compounds , and had no anti-inflammatory activity in the edema model induced by TPA.

Characterization of the Efflux Capability and Substrate Specificity of Aspergillus fumigatus PDR5-like ABC Transporters Expressed in Saccharomyces cerevisiae.

This research analyzed six genes encoding putative efflux proteins for their roles as transporters. Th genes , and were cloned into plasmids and overexpressed in a strain in which the highly active endogenous ABC transporter gene was deleted. The activity of each transporter was measured by efflux of rhodamine 6G and accumulation of alanine β-naphthylamide.

Pharmacogenomics at the center of precision medicine: challenges and perspective in an era of Big Data.

Pharmacogenomics (PGx) is one of the core elements of personalized medicine. PGx information reduces the likelihood of adverse drug reactions and optimizes therapeutic efficacy. St Catherine Specialty Hospital in Zagreb/Zabok, Croatia has implemented a personalized patient approach using the RightMed Comprehensive PGx panel of 25 pharmacogenes plus Facor V Leiden, Factor II and MTHFR genes, which is interpreted by a special counseling team to offer the best quality of care.

Production of distinct labdane-type diterpenoids using a novel cryptic labdane-like cluster from Streptomyces thermocarboxydus K155.

Bioinformatic mining of the Streptomyces thermocarboxydus K155 genome predicted the presence of four synthases for the production of geosmin, hopene, albaflavenone, and a type B-type A diterpenoid system like that described for labdane-related diterpenoids (LRD). The lrd cluster was comprised by an operon of four genes (lrdABDC). This cluster seemed to be silent in the wild-type strain, as neither labdane nor terpene-like compounds were detected by UPLC-TOF-MS and GC-MS analyses in both culture supernatants and mycelial extracts.

Exo-Metabolites of -Nodulating Rhizobial Strains.

Rhizobia are able to convert dinitrogen into biologically available forms of nitrogen through their symbiotic association with leguminous plants. This results in plant growth promotion, and also in conferring host resistance to different types of stress. These bacteria can interact with other organisms and survive in a wide range of environments, such as soil, rhizosphere, and inside roots.

The structure of (E)-biformene synthase provides insights into the biosynthesis of bacterial bicyclic labdane-related diterpenoids.

The labdane-related diterpenoids (LRDs) are a large group of natural products with a broad range of biological activities. They are synthesized through two consecutive reactions catalyzed by class II and I diterpene synthases (DTSs). The structural complexity of LRDs mainly depends on the catalytic activity of class I DTSs, which catalyze the formation of bicyclic to pentacyclic LRDs, using as a substrate the catalytic product of class II DTSs.

Author Correction: Drug-mediated metabolic tipping between antibiotic resistant states in a mixed-species community.

In the version of this Article originally published, the following sentence was missing from the Acknowledgements: "R.E.B.

Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in .

Ergosterol (ERG) is a critical sterol in the cell membranes of fungi, and its biosynthesis is tightly regulated by 25 known enzymes along the ERG production pathway. The effects of changes in expression of each ERG biosynthesis enzyme in were analyzed by the use of gene deletion or plasmid-borne overexpression constructs. The strains overexpressing the ERG pathway genes were examined for changes in doubling time and responses to a variety of stress agents.

Drug-mediated metabolic tipping between antibiotic resistant states in a mixed-species community.

Microbes rarely exist in isolation, rather, they form intricate multi-species communities that colonize our bodies and inserted medical devices. However, the efficacy of antimicrobials is measured in clinical laboratories exclusively using microbial monocultures. Here, to determine how multi-species interactions mediate selection for resistance during antibiotic treatment, particularly following drug withdrawal, we study a laboratory community consisting of two microbial pathogens.

Azole resistance in a mutant lacking the ABC transporter depends on TOR signaling.

ATP-binding cassette (ABC) transporters help export various substrates across the cell membrane and significantly contribute to drug resistance. However, a recent study reported an unusual case in which the loss of an ABC transporter in , orf19.4531 (previously named ROA1), increases resistance against antifungal azoles, which was attributed to an altered membrane potential in the mutant strain.

Structure, Absolute Configuration, and Antiproliferative Activity of Abietane and Icetexane Diterpenoids from Salvia ballotiflora.

From the aerial parts of , eleven diterpenoids were isolated; among them, four icetexanes and one abietane (-) are reported for the first time. Their structures were established by spectroscopic means, mainly ¹H- and C-NMR, including 1D and 2D homo- and hetero-nuclear experiments. Most of the isolated diterpenoids were tested for their antiproliferative, anti-inflammatory, and radical scavenging activities using the sulforhodamine B assay on six cancer cell lines, the TPA-induced ear edema test in mice, and the reduction of the DPPH assay, respectively.