Light and temperature effects on miR156 transgenic switchgrass flowering: A simulated latitudinal study.

The control of flowering in perennial grasses is an important trait, especially among biofuel feedstocks. Lignocellulosic biomass may be increased commensurate with decreased or delayed flowering as the plant allocates energy for stems and leaves harvested for bioenergy at the end of the growing season. For transgenic feedstocks, such as switchgrass (Panicum virgatum L.) grown in its geographic center of distribution, it is foreseeable that regulators may require greatly decreased gene flow frequencies to enable commercialization. Transgenic switchgrass with various overexpression levels of a rice microRNA gene, miR156, when grown in field conditions, holds promise for decreased flowering, yielding high biomass, and altered cell wall traits, which renders it as a potential crossing partner for further breeding with switchgrass lines for decreased recalcitrance. In the current research, we simulated a latitudinal cline in controlled growth chamber experiments for various individual sites from the tropics to cool-temperate conditions which included weekly average high and low temperatures and day lengths over the switchgrass growing season for each simulated site: Guayaquil, Ecuador; Laredo, Texas, USA; and Brattleboro, Vermont, USA. Flowering and reproduction among transgenic lines with low (T-14 and T-35)-to-moderate (T-27 and T-37) overexpression of miR156 were assessed. Lower simulated latitudes (higher temperatures with low-variant day length) and long growing seasons promoted flowering of the miR156 transgenic switchgrass lines. Tropical conditions rescued the flowering phenotype in all transgenic lines except T-27. Higher numbers of plants in lines T-35 and T-37 and the controls produced panicles, which also occurred earlier in the study as temperatures increased and day length decreased. Line T-14 was the exception as more clonal replicates flowered in the cool-temperate (Vermont) conditions. Increased biomass was found in transgenic lines T-35 and T-37 in tropical conditions. No difference in biomass was found in subtropical (Texas) chambers, and two lines (T-14 and T-35) produced less biomass than the control in cool-temperate conditions. Our findings suggest that switchgrass plants engineered to overexpress miR156 for delayed flowering to promote bioconfinement and biomass production may be used for plant breeding at tropical sites.

Multiple polyploidy events in the early radiation of nodulating and nonnodulating legumes.

Unresolved questions about evolution of the large and diverse legume family include the timing of polyploidy (whole-genome duplication; WGDs) relative to the origin of the major lineages within the Fabaceae and to the origin of symbiotic nitrogen fixation. Previous work has established that a WGD affects most lineages in the Papilionoideae and occurred sometime after the divergence of the papilionoid and mimosoid clades, but the exact timing has been unknown. The history of WGD has also not been established for legume lineages outside the Papilionoideae. We investigated the presence and timing of WGDs in the legumes by querying thousands of phylogenetic trees constructed from transcriptome and genome data from 20 diverse legumes and 17 outgroup species. The timing of duplications in the gene trees indicates that the papilionoid WGD occurred in the common ancestor of all papilionoids. The earliest diverging lineages of the Papilionoideae include both nodulating taxa, such as the genistoids (e.g., lupin), dalbergioids (e.g., peanut), phaseoloids (e.g., beans), and galegoids (=Hologalegina, e.g., clovers), and clades with nonnodulating taxa including Xanthocercis and Cladrastis (evaluated in this study). We also found evidence for several independent WGDs near the base of other major legume lineages, including the Mimosoideae-Cassiinae-Caesalpinieae (MCC), Detarieae, and Cercideae clades. Nodulation is found in the MCC and papilionoid clades, both of which experienced ancestral WGDs. However, there are numerous nonnodulating lineages in both clades, making it unclear whether the phylogenetic distribution of nodulation is due to independent gains or a single origin followed by multiple losses.

Comparison of a MEMS-Based Handheld OCT Scanner With a Commercial Desktop OCT System for Retinal Evaluation.

PURPOSE: The goal of this study was to evaluate the ability of our handheld optical coherence tomography (OCT) scanner to image the posterior and anterior structures of the human eye, and especially the individual layers of the retina, and to compare its diagnostic performance with that of a fixed desktop commercial ophthalmic OCT system. METHODS: We compared the clinical imaging results of our handheld OCT with a leading commercial desktop ophthalmic system (RTVue) used in specialist offices. Six patients exhibiting diabetes-related retinal pathology had both eyes imaged with each OCT system. RESULTS: In both sets of images, the structural irregularities of the retinal layers could be identified such as retinal edema and vitreomacular traction. CONCLUSIONS: Our handheld OCT system can be used to identify relevant anatomical structures and pathologies in the eye, potentially enabling earlier screening, disease detection, and treatment. Images can be acquired quickly, with sufficient resolution and negligible motion artifacts that would normally limit its diagnostic use. TRANSLATIONAL RELEVANCE: Following screening and early disease detection in primary care via our optimized handheld OCT system, patients can be referred to a specialist for treatment, preventing further disease progression. While many primary care physicians are adept at using the ophthalmoscope, they can definitely take advantage of more advanced technologies.

Altered lignin biosynthesis using biotechnology to improve lignocellulosic biofuel feedstocks.

Lignocellulosic feedstocks can be converted to biofuels, which can conceivably replace a large fraction of fossil fuels currently used for transformation. However, lignin, a prominent constituent of secondary cell walls, is an impediment to the conversion of cell walls to fuel: the recalcitrance problem. Biomass pretreatment for removing lignin is the most expensive step in the production of lignocellulosic biofuels. Even though we have learned a great deal about the biosynthesis of lignin, we do not fully understand its role in plant biology, which is needed for the rational design of engineered cell walls for lignocellulosic feedstocks. This review will recapitulate our knowledge of lignin biosynthesis and discuss how lignin has been modified and the consequences for the host plant.

Animal-Assisted Activity: Effects of a Complementary Intervention Program on Psychological and Physiological Variables.

Animal-assisted activity is the use of trained animals for the therapeutic, motivational, or educational benefit of patients. Subjects of this study were 218 patients hospitalized on the mental health unit of a community hospital with an existing, complementary animal-assisted activity program. Half of the patients participated in a 1-hour session of animal-assisted activity. The other half, who served as a comparison group, participated in a 1-hour stress management program. It was hypothesized that an animal-assisted activity program would improve ratings of depression, anxiety, and pain and the associated physiological measures of stress and discomfort. Self-report ratings of depression, anxiety, and pain were collected before and after treatment sessions, and blood pressure, pulse, and salivary cortisol were measured. There were significant decreases in depression (P < .0001), anxiety (P < .0001), pain (P < .0001), and pulse (P < .04) after animal-assisted activity program, comparable to those in the more traditional stress management group.

Overexpression of a soybean salicylic acid methyltransferase gene confers resistance to soybean cyst nematode.

Salicylic acid plays a critical role in activating plant defence responses after pathogen attack. Salicylic acid methyltransferase (SAMT) modulates the level of salicylic acid by converting salicylic acid to methyl salicylate. Here, we report that a SAMT gene from soybean (GmSAMT1) plays a role in soybean defence against soybean cyst nematode (Heterodera glycines Ichinohe, SCN). GmSAMT1 was identified as a candidate SCN defence-related gene in our previous analysis of soybean defence against SCN using GeneChip microarray experiments. The current study started with the isolation of the full-length cDNAs of GmSAMT1 from a SCN-resistant soybean line and from a SCN-susceptible soybean line. The two cDNAs encode proteins of identical sequences. The GmSAMT1 cDNA was expressed in Escherichia coli. Using in vitro enzyme assays, E. coli-expressed GmSAMT1 was confirmed to function as salicylic acid methyltransferase. The apparent Km value of GmSAMT1 for salicylic acid was approximately 46 µM. To determine the role of GmSAMT1 in soybean defence against SCN, transgenic hairy roots overexpressing GmSAMT1 were produced and tested for SCN resistance. Overexpression of GmSAMT1 in SCN-susceptible backgrounds significantly reduced the development of SCN, indicating that overexpression of GmSAMT1 in the transgenic hairy root system could confer resistance to SCN. Overexpression of GmSAMT1 in transgenic hairy roots was also found to affect the expression of selected genes involved in salicylic acid biosynthesis and salicylic acid signal transduction.

Noninvasive in vivo optical detection of biofilm in the human middle ear.

Otitis media (OM), a middle-ear infection, is the most common childhood illness treated by pediatricians. If inadequately treated, OM can result in long-term chronic problems persisting into adulthood. Children with chronic OM or recurrent OM often have conductive hearing loss and communication difficulties and require surgical treatment. Tympanostomy tube insertion, the placement of a small drainage tube in the tympanic membrane (TM), is the most common surgical procedure performed in children under general anesthesia. Recent clinical studies have shown evidence of a direct correspondence between chronic OM and the presence of a bacterial biofilm within the middle ear. Biofilms are typically very thin and cannot be recognized using a regular otoscope. Here we report the use of optical coherent ranging techniques to noninvasively assess the middle ear to detect and quantify biofilm microstructure. This study involves adults with chronic OM, which is generally accepted as a biofilm-related disease. Based on more than 18,537 optical ranging scans and 742 images from 13 clinically infected patients and 7 normal controls using clinical findings as the gold standard, all middle ears with chronic OM showed evidence of biofilms, and all normal ears did not. Information on the presence of a biofilm, along with its structure and response to antibiotic treatment, will not only provide a better fundamental understanding of biofilm formation, growth, and eradication in the middle ear, but also may provide much-needed quantifiable data to enable early detection and quantitative longitudinal treatment monitoring of middle-ear biofilms responsible for chronic OM.

Switchgrass (Panicum virgatum L.) polyubiquitin gene (PvUbi1 and PvUbi2) promoters for use in plant transformation.

BACKGROUND: The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (Panicum virgatum L.) ubiquitin genes (PvUbi1 and PvUbi2) were cloned and characterized. Reporter constructs were produced containing the isolated 5' upstream regulatory regions of the coding sequences (i.e. PvUbi1 and PvUbi2 promoters) fused to the uidA coding region (GUS) and tested for transient and stable expression in a variety of plant species and tissues. RESULTS: PvUbi1 consists of 607 bp containing cis-acting regulatory elements, a 5' untranslated region (UTR) containing a 93 bp non-coding exon and a 1291 bp intron, and a 918 bp open reading frame (ORF) that encodes four tandem, head -to-tail ubiquitin monomer repeats followed by a 191 bp 3' UTR. PvUbi2 consists of 692 bp containing cis-acting regulatory elements, a 5' UTR containing a 97 bp non-coding exon and a 1072 bp intron, a 1146 bp ORF that encodes five tandem ubiquitin monomer repeats and a 183 bp 3' UTR. PvUbi1 and PvUbi2 were expressed in all examined switchgrass tissues as measured by qRT-PCR. Using biolistic bombardment, PvUbi1 and PvUbi2 promoters showed strong expression in switchgrass and rice callus, equaling or surpassing the expression levels of the CaMV 35S, 2x35S, ZmUbi1, and OsAct1 promoters. GUS staining following stable transformation in rice demonstrated that the PvUbi1 and PvUbi2 promoters drove expression in all examined tissues. When stably transformed into tobacco (Nicotiana tabacum), the PvUbi2+3 and PvUbi2+9 promoter fusion variants showed expression in vascular and reproductive tissues. CONCLUSIONS: The PvUbi1 and PvUbi2 promoters drive expression in switchgrass, rice and tobacco and are strong constitutive promoter candidates that will be useful in genetic transformation of monocots and dicots.

Antimicrobial activity of Yerba Mate (Ilex paraguariensis) aqueous extracts against Escherichia coli O157:H7 and Staphylococcus aureus.

UNLABELLED: Bioactive compounds from natural plant sources are becoming increasingly important to the food industry. Ilex paraguariensis is used in the preparation of a widely popular tea beverage (Yerba Mate) in the countries of Uruguay, Paraguay, Argentina, and Brazil. In this study, extracts of 4 brands of commercial tea, derived from the holly plant species, Ilex paraguariensis, were evaluated for their ability to inhibit or inactivate bacterial foodborne pathogens. The ultimate goal was to evaluate potential use of the extracts in commercial applications. Dialyzed aqueous extracts were screened for antimicrobial activity against Escherichia coli O157:H7 and Staphylococcus aureus. S. aureus was found to be the more sensitive to extracts than E. coli O157:H7. Minimum bactericidal concentrations (MBCs) were determined to be approximately 150 to 800 µg/mL and 25 to 50 µg/mL against E. coli O157:H7 and S. aureus, respectively. A Uruguayan brand had reduced activity against E. coli O157:H7 compared to the Argentinean brands tested. It was concluded that Yerba Mate could be used as a potential antimicrobial in foods and beverages against these pathogenic bacteria. PRACTICAL APPLICATION: Soluble extracts from Yerba Mate are natural antimicrobials that can be incorporated into food products to achieve longer shelf life.

Handheld optical coherence tomography scanner for primary care diagnostics.

The goal of this study is to develop an advanced point-of-care diagnostic instrument for use in a primary care office using handheld optical coherence tomography (OCT). This system has the potential to enable earlier detection of diseases and accurate image-based diagnostics. Our system was designed to be compact, portable, user-friendly, and fast, making it well suited for the primary care office setting. The unique feature of our system is a versatile handheld OCT imaging scanner which consists of a pair of computer-controlled galvanometer-mounted mirrors, interchangeable lens mounts, and miniaturized video camera. This handheld scanner has the capability to guide the physician in real time for finding suspicious regions to be imaged by OCT. In order to evaluate the performance and use of the handheld OCT scanner, the anterior chamber of a rat eye and in vivo human retina, cornea, skin, and tympanic membrane were imaged. Based on this feasibility study, we believe that this new type of handheld OCT device and system has the potential to be an efficient point-of-care imaging tool in primary care medicine.

Non-invasive optical interferometry for the assessment of biofilm growth in the middle ear.

Otitis media (OM) is the most common illness in children in the United States. Three-fourths of children under the age of three have OM at least once. Children with chronic OM, including OM with effusion and recurrent OM, will often have conductive hearing loss and communication difficulties, and need surgical treatment. Recent clinical studies provide evidence that almost all chronic OM cases are accompanied by a bacterial biofilm behind the tympanic membrane (eardrum) and within the middle ear. Biofilms are typically very thin, and cannot be recognized using a regular otoscope. Here we demonstrate how optical low coherence interferometry (LCI) noninvasively depth-ranges into the middle ear to detect and quantify biofilm microstructure. A portable diagnostic system integrating LCI with a standard video otoscope was constructed and used to detect and quantify the presence of biofilms in a newly-developed pre-clinical animal model for this condition. Using a novel classification algorithm for acquired LCI data, the system identified the presence of a biofilm with 86% sensitivity and 90% specificity, compared to histological findings. This new information on the presence of a biofilm, its structure, and its response to antibiotic treatment, will not only provide better understanding of fundamental principles that govern biofilm formation, growth, and eradication, but may also provide much needed clinical data to direct and monitor protocols for the successful management of otitis media.