Evaluation of growth and carotenoid production by a green microalga Scenedesmus quadricauda PUMCC 4.1.40. under optimized culture conditions.

Microalgae are a potential source of a wide range of food and novel value-added products. The versatility of microalgae to produce different kind of pigments is gaining interest as a sustainable source of natural carotenoids. Currently, commercial production of carotenoids from selected microalgae requires special culture conditions which are difficult to maintain. The present study has been undertaken to optimize culture conditions for growth and carotenoid production by a new isolate Scenedesmus quadricauda PUMCC 4.1.40. The results revealed that test organism produced 1.54 mg dry biomass/ml with a content of 40 µg carotenoids/mg dry biomass during stationary phase. The growth and carotenoid production was increased by 2.4-fold under combined optimized culture conditions. The optimized conditions were growth medium, Chu-10; pH 8.5; temperature, 30°C; nitrogen, 20 mM nitrate; phosphate, 0.22 mM; NaCl, 0.42 mM and blue light. Separation and identification of four important carotenoids through high-performance thin-layer chromatography (HPTLC) followed by purification using flash chromatography and quantification by HPLC revealed 23.8, 19.0, 6.5, and 4.0 µg astaxanthin, ß-carotene, lutein, and canthaxanthin /mg dry biomass, respectively. The amount of total carotenoids (98 µg/mg dry biomass) containing 40% valuable astaxanthin and ß-carotene produced under optimized conditions was significantly higher than control cultures. This justifies that S. quadricauda is a promising candidate for scale-up production of carotenoid.

Returning genomic results in a Federally Qualified Health Center: the intersection of precision medicine and social determinants of health.

PURPOSE: This report describes the return of sequencing results to low-income Latino participants recruited through a Federally Qualified Health Center (FQHC). We describe challenges in returning research results secondary to social determinants of health and present lessons learned to guide future genomic medicine implementation studies in low-resource settings. METHODS: Five hundred Latino adults (76% women) consented to research sequencing for a predetermined panel of actionable genes. Providers and staff from the FQHC were engaged to align processes with the practice and a community advisory board grounded the project in the local community. RESULTS: A pathogenic/likely pathogenic variant was present in 10 participants (2%). Challenges in return of results included the time lag (582 ± 53 days) between enrollment and returning actionable results, difficulty reaching participants, missed appointments, low health literacy, lack of health insurance, and reconciling results with limited information on family history. Return of one actionable result was deferred due to acute emotional distress secondary to recent traumatic life events. CONCLUSION: The social determinants of health influence the implementation of genomic medicine in low-income populations in low-resource settings. Considering nonbiological factors that contribute to disparities will be necessary to better appreciate how genomic medicine may fit within the context of health equity.

Improving Access to Care Through the Establishment of a Local, Teledermatology Network.

Introduction: Access to dermatologic care is a major issue in the United States, especially within the un- and underinsured populations; technology, including teledermatology, will pay a role in improving access to care. Methods: We performed a prospective study between November 2016 and September 2017. We leveraged a partnership between Mayo Clinic and Mountain Park Health Clinic, a community clinic that primarily serves un- and underinsured populations. We implemented a mobile phone-based store and forward (SAF) teledermatology service, which integrated an external community health clinic to an existing electronic health record (EHR) using standardized data capture forms, real-time support, and simple workflows. Results: Thirty-seven patients were enrolled in the study, 65% female and 35% male with an average age of 47.9 (SD = 15.9). The ethnic breakdown was: 81.1% Hispanic, 13.5% Caucasian, and 5.4% African American. The majority, 62.2%, did not have a high school education, 45.9% were unemployed, and 51.4% were uninsured. 64.9% earned less than $25,000 for annual household income. Teledermatology consultation increased the absolute diagnostic and management concordance by 36.6% (p = 0.01, 95% CI 12.2%-61.0%) and 34.2% (p < 0.01, 95% CI 11%-57%), respectively. Primary care providers had a significant increase in mean confidence in the diagnosis and management of dermatology conditions pre and poststudy (3.60 vs. 3.70 and 3.21 vs. 3.60, respectively; p < 0.01). Ninety-six percent of the primary care providers agreed (52.0%) and strongly agreed (44.0%) that they would send another patient for teleconsultation.Conclusion: We successfully implemented a SAF teledermatology consultative service in a community health clinic outside our EHR. A similar approach can be used by other large health care organizations to provide integrated, high-quality consultation to clinics with rural, un- and underinsured populations.

Developing a Process for Returning Medically Actionable Genomic Variants to Latino Patients in a Federally Qualified Health Center.

AIM: To develop a process for returning medically actionable genomic variants to Latino patients receiving care in a Federally Qualified Health Center. METHODS: Prior to recruitment, researchers met with primary care providers to (1) orient clinicians to the project, (2) establish a process for returning actionable and nonactionable results to participants and providers through the electronic health record, and (3) develop a process for offering clinical decision support for follow-up education and care. A Community Advisory Board was engaged to provide input on recruitment strategies and materials for conveying results to participants. Participants in the Sangre Por Salud (Blood for Health) Biobank with hyperlipidemia or colon polyps represented the pool of potentially eligible participants. RESULTS: A total of 1,621 individuals were invited to participate and 710 agreed to an in- person consenting visit (194 no-showed and 16 declined). Over 12-months, 500 participants were enrolled. Participants were primarily Spanish-speaking (81.6%), female (74.2%), and enrolled because of hyperlipidemia (95.4%). All but 2 participants opted to receive primary (i.e., related to enrollment phenotypes) as well as secondary actionable results. CONCLUSION: Efforts to bring precision medicine to community-based health centers serving minority patients may require multilevel engagement activities to include individuals, providers, health systems, and the community.

Exposure of grapes to smoke of vegetation with varying lignin composition and accretion of lignin derived putative smoke taint compounds in wine.

Smoke taint in wines from bushfire smoke exposure has become a concern for wine producers. Smoke taint compounds are primarily derived from pyrolysis of the lignin component of fuels. This work examined the influence of the lignin composition of pyrolysed vegetation on the types of putative smoke taint compounds that accrue in wines. At veraison, Merlot vines were exposed to smoke generated from five vegetation types with differing lignin composition. Smoke was generated under pyrolysis conditions that simulated bushfire temperature profiles. Lignin and smoke composition of each fuel type along with putative smoke taint compounds in wines were determined. The results showed that, regardless of fuel type, the commonly reported guaiacyl lignin derived smoke taint compounds, guaiacol and 4-methylguaiacol, represented about 20% of the total phenols in wines. Quantitatively, syringyl lignin derived compounds dominated the total phenol pools in both free and bound forms. The contributions of p-hydroxyphenyls were generally similar to the guaiacyl sources. A further unexpected outcome of the study was that pine smoke affected wines had significantly elevated levels of syringols compared to the controls although pine fuel and its smoke emission lacked syringyl products.

Impact of boron, calcium and genetic factors on vitamin C, carotenoids, phenolic acids, anthocyanins and antioxidant capacity of carrots (Daucus carota).

Carrots (Daucus carota L.) were used to investigate the effects and interactions of cultivar and mineral supply on the nutritional quality (antioxidant potential, vitamin C, carotenoids and phenolic acids) of the resulting storage roots. The supplement of boron (B) and or calcium (Ca) in the feeding solutions, during plant growth, influenced the accumulation of other minerals, such as P, K, Mg, S and Na, in the storage roots (p<0.05). When no additional B or Ca was supplied (e.g. -B or -Ca treatment), we observed 33-50% increase in the accumulated levels of α- and ß-carotenes, and 45-70% increase of vitamin C. Carrots grown with no supplement of B in the nutrient solutions (e.g. -B treatment and -ve control) had significantly higher (p<0.001) levels of total phenolic acids compared to the carrots with the supplement of B (e.g. -Ca treatment and +ve control). A strong positive correlation was observed between the total phenolic contents and ORAC values (r=0.932) in all the cultivars. The results suggest that both cultivar and mineral supply were major determinants of nutritional quality of the carrots. The nutritional value of carrot crops (with an acceptable physical quality) can be enhanced by manipulating mineral nutrient applications.

Micronutrient mineral and folate content of Australian and imported dried fruit products.

A selection of Australian and imported fresh and dried fruit products, including sultanas, Sunmuscats, Carina currants, Zante currants, apricots, and prunes, were analyzed for selected minerals (Ca, Mg, Na, S, B, Al, Fe, Mn, Cu, Zn, Mo, and Se), folate and vitamin C, and the capacity of dried fruits for dietary provision of these micronutrients evaluated. Micro-nutrients were concentrated by a factor of 3-5 in dried fruits compared with their fresh fruit counterparts and were consequently present in nutritionally significant levels, in contrast to fresh fruit. Australian dried sultanas, Carina currant, Zante currant, apricots, and prunes contained Cu, Fe, K, and Mn at levels of >20% of daily Required Dietary Intake (RDI, taken as the average for adult men and women as nominated by the Australian National Health and Medical Research Council) and Sunmuscats contained Cu, Fe, and K at >20% of RDI. All dried fruits studied contained boron in the range of 1.5 to 5.4 mg per 100 g; however, the RDI for boron has not been defined by the NHMRC at the present time. All sultanas and currants studied contained folate at levels of 10-20% of RDI per 100 g. Experimental drying methods significantly affected folate levels with higher folate content in non-ground versus ground-based drying methods. Of the micro-nutrients supplying >20% of RDI, folate represents a particular nutrient for which the mean daily intake of adult Australians is typically inadequate. This study shows that dried fruit consumption, in contrast with fresh fruit, can provide significant proportions of daily requirements of several micronutrients, particularly folate.

The microbes of the intestine: an introduction to their metabolic and signaling capabilities.

This article summarizes advances in the field of host-microbe interactions in the gut. The human gut is home to a complex community of microbes (the microbiota) that plays a critical role in host nutrient acquisition and metabolism, development of intestinal epithelial cells, and host immune system. Genetic background, nutritional status, and environmental factors influence the structure and function of the gut microbiota. Networks for cell-cell communication include microbes actively communicating with microbes of the same and other species; host cells recognizing and interacting with commensal versus pathogenic organisms; and microbes releasing peptides that resemble peptide hormones of vertebrates, possibly influencing host cell function.

Tall tales from sly dwarves: novel functions of gibberellins in plant development.

Gibberellins (GAs) are endogenous hormones controlling numerous aspects of plant growth and development. Our present understanding of GA physiology is based largely on genetic analysis in model plants such as Arabidopsis. In spite of the success of this approach, the discovery of additional physiological roles for GAs in seed development, pollen tube growth and meristem development indicates that the existing collection of GA-related mutants (identified partially or entirely on the basis of vegetative phenotypes) has failed to uncover all aspects of plant development that are controlled by GAs. The continued use of ever improving forward and reverse genetic techniques is expected to lead to the discovery of further novel roles for GAs in plant development.

Plants with increased expression of ent-kaurene oxidase are resistant to chemical inhibitors of this gibberellin biosynthesis enzyme.

The gibberellin (GA) biosynthetic pathway includes the three-step oxidation of ent-kaurene to ent-kaurenoic acid, catalyzed by the enzyme ent-kaurene oxidase (KO). Arabidopsis plants overexpressing the KO cDNA under the control of the cauliflower mosaic virus 35S promoter, with or without a translational fusion to a modified green fluorescent protein (GFP), are very similar to wild-type (WT) plants under normal growth conditions. In contrast, when WT and 35S:KO (or 35S:KO-GFP) seeds, seedlings or pollen tubes are grown in the presence of chemical inhibitors of KO, such as paclobutrazol and uniconazole, plants with increased KO expression are partially resistant to the effects of these inhibitors. In combination with the observation that decreased KO levels increase the sensitivity to KO inhibitors, the 35S:KO phenotypes demonstrate that the modification of KO enzyme levels could be used to create transgenic crop plants with altered KO inhibitor response. These results also suggest that the KO gene could be used as a selectable marker for plant regeneration based on resistance to KO inhibitors. Finally, the observation that pollen tubes expressing 35S:KO or 35S:KO-GFP have decreased sensitivity to KO inhibitors provides further evidence for a physiological role for GAs in pollen tube elongation.

The gar2 and rga alleles increase the growth of gibberellin-deficient pollen tubes in Arabidopsis.

Ectopic expression in Arabidopsis of a pea (Pisum sativum) cDNA (2ox2) encoding a gibberellin (GA) 2-oxidase (PsGA2ox2), involved in the deactivation of biologically active GAs, has been used to establish a role for GAs in promoting pollen tube growth. One line, 35S:2ox2/28c, when homozygous for the transgene, exhibits a novel small fruit phenotype. The 28c transgene reduces pollen tube growth, and this results in a reduced number of fertilized seeds that are only present at the end of the silique nearest the stigma. To confirm that the 28c pollen tube phenotype is due to sense expression of the 2ox2 mRNA, a "hairpin" RNA interface silencing construct, designed to silence 2ox2 expression, has been used to restore pollen tube growth and fruit development. The interaction between 28c and other mutants with increased GA response has also been examined to provide further evidence that GAs play an important role in pollen tube growth. Based on the ability of mutant alleles to suppress the 35S:2ox2/28c phenotype, we define new roles for the gar2-1 and rga alleles in GA signaling during pollen tube elongation in addition to their previously established roles in vegetative tissues. In contrast to the constitutive GA response observed in internodes and leaves lacking RGA and GAI, the rga-2 gai-d5 mutant combination is only a partial suppressor of the 28c phenotype. Because the dominant dwarfing gai-1 allele reduces GA response in vegetative tissues, its effect on plant fertility has been examined. Although gai-1 reduces seed set, this appears to reflect defects in reproductive development other than pollen tube function. Finally, we show that the genetic background (Landsberg erecta or Columbia) modifies the 28c phenotype and that this effect is not due to the ER/er difference between these two ecotypes.

Activation of multiple antiviral defence mechanisms by salicylic acid.

SUMMARY The plant signal molecule salicylic acid (SA) can induce resistance to a wide range of pathogen types. In the case of viruses, SA can stimulate the inhibition of all three main stages in virus infection: replication, cell-to-cell movement and long-distance movement. Induction of resistance by SA appears to depend, in part, on downstream signalling via the mitochondrion. However, evidence has recently emerged that SA may stimulate a separate downstream pathway, leading to the induction of an additional mechanism of resistance based on RNA interference. In this review our aims are to document these recent advances and to suggest possible future avenues of research on SA-induced resistance to viruses.

Genetic modification of alternative respiration has differential effects on antimycin A-induced versus salicylic acid-induced resistance to Tobacco mosaic virus.

Salicylic acid (SA), a natural defensive signal chemical, and antimycin A, a cytochrome pathway inhibitor, induce resistance to Tobacco mosaic virus (TMV). Pharmacological evidence suggested signaling during resistance induction by both chemicals involved alternative oxidase (AOX), sole component of the alternative respiratory pathway (AP). Roles of the AP include regulation of intramitochondrial reactive oxygen species and maintenance of metabolic homeostasis. Transgenic tobacco (Nicotiana tabacum) with modified AP capacities (2- to 3-fold increased or decreased) showed no alteration in phenotype with respect to basal susceptibility to TMV or the ability to display SA-induced resistance to systemic viral disease. However, in directly inoculated tissue, antimycin A-induced TMV resistance was inhibited in plants with increased AP capacities, whereas SA and antimycin A-induced resistance was transiently enhanced in plant lines with decreased AP capacities. We conclude that SA-induced TMV resistance results from activation of multiple mechanisms, a subset of which are inducible by antimycin A and influenced by AOX. Other antiviral factors, potentially including the SA-inducible RNA-dependent RNA polymerase, are regulated by AOX-independent mechanisms.

Gibberellins are required for seed development and pollen tube growth in Arabidopsis.

Gibberellins (GAs) are tetracyclic diterpenoids that are essential endogenous regulators of plant growth and development. GA levels within the plant are regulated by a homeostatic mechanism that includes changes in the expression of a family of GA-inactivating enzymes known as GA 2-oxidases. Ectopic expression of a pea GA 2-oxidase2 cDNA caused seed abortion in Arabidopsis, extending and confirming previous observations obtained with GA-deficient mutants of pea, suggesting that GAs have an essential role in seed development. A new physiological role for GAs in pollen tube growth in vivo also has been identified. The growth of pollen tubes carrying the 35S:2ox2 transgene was reduced relative to that of nontransgenic pollen, and this phenotype could be reversed partially by GA application in vitro or by combining with spy-5, a mutation that increases GA response. Treatment of wild-type pollen tubes with an inhibitor of GA biosynthesis in vitro also suggested that GAs are required for normal pollen tube growth. These results extend the known physiological roles of GAs in Arabidopsis development and suggest that GAs are required for normal pollen tube growth, a physiological role for GAs that has not been established previously.