Metabolomic response of Perilla frutescens leaves, an edible-medicinal herb, to acclimatize magnesium oversupply.

High salt accumulation, resulting from the rampant use of chemical fertilizers in greenhouse cultivation, has deleterious effects on plant growth and crop yield. Herein, we delineated the effects of magnesium (Mg) oversupply on Perilla frutescens leaves, a traditional edible and medicinal herb used in East-Asian countries. Mg oversupply resulted in significantly higher chlorophyll content coupled with lower antioxidant activities and growth, suggesting a direct effect on subtle metabolomes. The relative abundance of bioactive phytochemicals, such as triterpenoids, flavonoids, and cinnamic acids, was lower in the Mg-oversupplied plants than in the control. Correlation analysis between plant phenotypes (plant height, total fresh weight of the shoot, leaf chlorophyll content, and leaf antioxidant content) and the altered metabolomes in P. frutescens leaves suggested an acclimatization mechanism to Mg oversupply. In conclusion, P. frutescens preferentially accumulated compatible solutes, i.e., carbohydrates and amino acids, to cope with higher environmental Mg levels, instead of employing secondary and antioxidative metabolism.

Effects of diluted bitumen exposure and recovery on the seawater acclimation response of Atlantic salmon smolts.

Petrogenic chemicals are common and widespread contaminants in the aquatic environment. In Canada, increased extraction of bitumen from the oil sands and transport of the major crude oil export product, diluted bitumen (dilbit), amplifies the risk of a spill and contamination of Canadian waterways. Fish exposed to sublethal concentrations of crude oil can experience a variety of adverse physiological effects including osmoregulatory dysfunction. As regulation of water and ion balance is crucial during the seawater transition of anadromous fish, the hypothesis that dilbit impairs seawater acclimation in Atlantic salmon smolts (a fish at risk of exposure in Canada) was tested. Smolts were exposed for 24 d to the water-soluble fraction of dilbit in freshwater, and then transferred directly to seawater or allowed a 1 wk depuration period in uncontaminated freshwater prior to seawater transfer. The seawater acclimation response was quantified at 1 and 7 d post-transfer using established hematological, tissue, and molecular endpoints including gill Na+/K+-ATPase gene expression (nka). All smolts, irrespective of dilbit exposure, increased serum Na+ concentrations and osmolality within 1 d of seawater transfer. The recovery of these parameters to freshwater values by 7 d post-transfer was likely driven by the increased expression and activity of Na+/K+-ATPase in the gill. Histopathological changes in the gill were not observed; however, CYP1A-like immunoreactivity was detected in the pillar cells of gill lamellae of fish exposed to 67.9 µg/L PAC. Concentration-specific changes in kidney expression of a transmembrane water channel, aquaporin 3, occurred during seawater acclimation, but were resolved with 1 wk of depuration and were not associated with histopathological changes. In conclusion, apart from a robust CYP response in the gill, dilbit exposure did not greatly impact common measures of seawater acclimation, suggesting that significant osmoregulatory dysfunction is unlikely to occur if Atlantic salmon smolts are exposed sub-chronically to dilbit.

Efficacy of Shenqi Pollen Capsules for High-Altitude Deacclimatization Syndrome via Suppression of the Reoxygenation Injury and Inflammatory Response.

High-altitude deacclimatization syndrome (HADAS) is involved in hypoxia-reoxygenation injury and inflammatory response, induced a series of symptoms, and has emerged as a severe public health issue. Here, we investigated the mechanism as well as potential means to prevent HADAS using Shenqi pollen capsules (SPCs) in subjects with HADAS in a multicenter, double-blinded, randomized, placebo-controlled study. All subjects were at the same high altitude (3650 m) for 4-8 months before returning to lower altitudes. Subjects (n = 288) in 20 clusters were diagnosed with mild or moderate HADAS on the third day of the study. We randomly allocated 20 clusters of subjects (1 : 1) to receive SPCs or a placebo for 7 weeks, and they were then followed up to the 14th week. The primary endpoints were subjects' HADAS scores recorded during the 14 weeks of follow-up. Compared with the placebo, SPC treatment significantly decreased the subjects' HADAS scores and reduced the incidence of symptom persistence. SPC therapy also reduced the serum levels of CK, CK-MB, LDH, IL-17A, TNF-α, and miR-155 and elevated IL-10 and miR-21 levels. We thus demonstrate that SPCs effectively ameliorated HADAS symptoms in these subjects via suppression of the hypoxia-reoxygenation injury and inflammatory response.

Indirect Regeneration and Assessment of Genetic Fidelity of Acclimated Plantlets by SCoT, ISSR, and RAPD Markers in Rauwolfia tetraphylla L.: An Endangered Medicinal Plant.

Rauwolfia tetraphylla L. is an important medicinal plant species which is well known for its pharmaceutically important alkaloids. In the present study, we are reporting about its conservation by in vitro clonal multiplication through the standardized protocol of indirect regeneration by using leaf and stem based callus and assessment of genetic fidelity of acclimated plantlets by start codon targeted (SCoT), inter simple sequence repeats (ISSR), and randomly amplified polymorphic DNA (RAPD) marker based analysis. Initially friable callus was induced in maximum amounts (378.7, 323.8, and 412.8 in mg) from leaf, root, and stem explants on Murashige and Skoog (MS) media supplemented with 5.0 mg/L, 3.0 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.0 mg/L of naphthalene acetic acid (NAA), respectively. Shoot regeneration with the maximum number of shoot buds (25 and 20) was obtained from leaf and stem calluses on MS media supplemented with TDZ (0.25 mg/L) + BAP (2 mg/L). The regenerated shoots were rooted successfully with maximum rooting percentage of 98.0 on full strength MS media amended with IAA (1.0 mg/L) and IBA (1.0 mg/L). The regenerated plantlets were hardened using 2:1 ratio of sterile garden soil and sand, followed by acclimatization in field conditions with 86% of survival. SCoT, ISSR, and RAPD primers based polymerase chain reaction (PCR) analysis was carried out to check possible genetic variations in micro propagated plants in comparison with mother plant. Among the ten SCoT (S), ISSR (R), and RAPD (OPA) primers used, S2, R10, and OPA3 has given good amplification with scorable DNA bands. The results revealed that the regenerated plants did not have any polymorphism with mother plant. Hence, the in vitro regenerated R. tetraphylla plantlets were confirmed as true-to-type.

Trans-Ferulic Acid-4-ß-Glucoside Alleviates Cold-Induced Oxidative Stress and Promotes Cold Tolerance.

Trans-ferulic acid-4-ß-glucoside (C16H20O9, TFA-4ß-G) is a monomer extracted from the Chinese medicine called radix aconiti carmichaeli (Fuzi). To date, research on this substance is lacking. Here, we found that trans-ferulic acid-4-ß-glucoside effectively promoted cold acclimatization in mice via increased heat production and alleviation of oxidative stress in a cold environment. Thus, our work indicates that ferulic acid-4-ß-glucoside is a potential therapeutic candidate for prevention and treatment of cold stress injury.

Production of lutein, and polyunsaturated fatty acids by the acidophilic eukaryotic microalga Coccomyxa onubensis under abiotic stress by salt or ultraviolet light.

In this study, the effect of abiotic stress on the acidophilic eukaryotic microalga, Coccomyxa onubensis, was analyzed for the production of lutein and PUFAs (polyunsaturated fatty acids). It grows autotrophically at a pH of 2.5. It showed a growth rate of 0.30 d-1, and produced approximately 122.50 mg·L-1·d-1 biomass, containing lipids (300.39 mg g-1dw), lutein (5.30 mg g-1dw), and ß-carotene (1.20 mg g-1dw). The fatty acid methyl ester (FAME) fraction was 89.70 mg g-1dw with abundant palmitic acid (28.70%) and linoleic acid (37.80%). The addition of 100 mM NaCl improved the growth rate (0.54 d-1), biomass productivity (243.75 mg·L-1·d-1), and lipids accumulation (416.16 mg g-1dw). The microalga showed a lutein content of 6.70 mg g-1dw and FAME fraction of 118.90 mg g-1dw; 68% of the FAMEs were PUFAs. However, when 200-500 mM salt was added, its growth was inhibited but there was a significant induction of lutein (up to 7.80 mg g-1dw). Under continuous illumination with PAR (photosynthetically active radiations) +UVA (ultraviolet A, 8.7 W m-2), C. onubensis showed a growth rate of 0.40 d-1, and produced 226.3 mg·L-1·d-1 biomass, containing lipids, (487.26 mg g-1dw), lutein (7.07 mg g-1dw), and FAMEs (232.9 mg g-1dw); 48.4% of the FAME were PUFAs. The illumination with PAR + UVB (ultraviolet B, 0.16 W m-2) was toxic for cells. These results indicate that C. onubensis biomass is suitable as a supplement for functional foods and/or source of high added value products.

Selenium nanoparticles enhanced thermal tolerance and maintain cellular stress protection of Pangasius hypophthalmus reared under lead and high temperature.

There is strong relation between nutrition and thermal tolerance of fish in terms of improved critical temperature minima (CTmin), lethal temperature minima (LTmin), critical temperature maxima (CTmax), and lethal temperature maxima (LTmax). Fishes act as quantifying indicators of the climate change due to their critical thermal limits in nature and ability to adjust thermal sensitivity to maintain their homeostasis. In the present study, biologically synthesised selenium nanoparticles (Se-NPs) from fisheries waste (fish gill) were used to evaluate its efficacy for enhancement of thermal tolerance of Pangasius hypophthalmus reared under lead (Pb) and high temperature (34°C) for 72days. At the end of the experimental period, CTmin, LTmin, and CTmax, LTmax and cellular metabolic stress were evaluated. The CTmin, LTmin were noticeably (p<0.01) reduced and CTmax, LTmax were enhanced in treatment groups fed with dietary Se-NPs. It was also observed that CTmax and LTmax were increased with acclimation temperature. The positive correlation was perceived between CTmin, LTmin (Y=-0.45+11.05x, R2, 0.768) and CTmax, LTmax (Y=-0.78+8.17x, R2, 0.858) fed with Se-NPs supplemented diet and exposed to Pb and high temperature. Significant improvements in the oxidative and metabolic enzymes were noticed post thermal tolerance experiment. Overall results indicated that incorporation of Se-NPs @ 1mg/kg in diet can confer protection against Pb and thermal stress in P. hypophthalmus.

Influence of crude oil exposure on cardiac function and thermal tolerance of juvenile rainbow trout and European sea bass.

Oil spills pose a threat to aquatic organisms. However, the physiological effects of crude oil on cardiac function and on thermal tolerance of juvenile fish are still poorly understood. Consequently, in this paper, we will present results of two separate experiments where we exposed juvenile rainbow trout and European sea bass to crude oil and made cardiac thermal tolerances and maximum heart rate (f Hmax) measurements after 1 week (rainbow trout) and 6-month recovery (sea bass). In both species, the f Hmax was lower in crude oil-exposed fish than in the control ones at temperatures below the optimum but this difference disappeared at higher temperatures. More importantly, the oil-exposed fish had significantly higher Arrhenius break point temperature for f Hmax, which gave an estimate for optimum temperature, than the control fish in both species even though the exposure conditions and recovery times differed between species. The results indicated that exposure of juvenile fish to crude oil did not have a significant negative impact upon their cardiac performance in high temperatures and upper thermal tolerance increased when the fish were tested 1 week or 6 months after the exposure. Our findings suggest that the cardiac function and thermal tolerance of juvenile fish are relatively resistant to a crude oil exposure.

Combinatorial therapy of exercise-preconditioning and nanocurcumin formulation supplementation improves cardiac adaptation under hypobaric hypoxia.

BACKGROUND: Chronic hypobaric hypoxia (cHH) mediated cardiac insufficiencies are associated with pathological damage. Sustained redox stress and work load are major causative agents of cardiac insufficiencies under cHH. Despite the advancements made in pharmacological (anti-oxidants, vasodilators) and non-pharmacological therapeutics (acclimatization strategies and schedules), only partial success has been achieved in improving cardiac acclimatization to cHH. This necessitates the need for potent combinatorial therapies to improve cardiac acclimatization at high altitudes. We hypothesize that a combinatorial therapy comprising preconditioning to mild aerobic treadmill exercise and supplementation with nanocurcumin formulation (NCF) consisting of nanocurcumin (NC) and pyrroloquinoline quinone (PQQ) might improve cardiac adaptation at high altitudes. METHODS: Adult Sprague-Dawley rats pre-conditioned to treadmill exercise and supplemented with NCF were exposed to cHH (7620 m altitude corresponding to pO2~8% at 28±2°C, relative humidity 55%±1%) for 3 weeks. The rat hearts were analyzed for changes in markers of oxidative stress (free radical leakage, lipid peroxidation, manganese-superoxide dismutase [MnSOD] activity), cardiac injury (circulating cardiac troponin I [TnI] and T [cTnT], myocardial creatine kinase [CK-MB]), metabolic damage (lactate dehydrogenase [LDH] and acetyl-coenzyme A levels, lactate and pyruvate levels) and bio-energetic insufficiency (ATP, p-AMPKα). RESULTS: Significant modulations (p≤0.05) in cardiac redox status, metabolic damage, cardiac injury and bio-energetics were observed in rats receiving both NCF supplementation and treadmill exercise-preconditioning compared with rats receiving only one of the treatments. CONCLUSIONS: The combinatorial therapeutic strategy showed a tremendous improvement in cardiac acclimatization to cHH compared to either exercise-preconditioning or NCF supplementation alone which was evident from the effective modulation in redox, metabolic, contractile and bio-energetic homeostasis.

Cold adaptive thermogenesis following consumption of certain pungent spice principles: A validation study.

Identifying a means to activate or potentiate thermogenic mechanisms through ingestion of dietary compounds have important implications in cold endurance and survival. Although many reports discuss the thermogenic potential of spices, it is surprising that none of the studies verify whether consumption of spices can improve cold endurance. In this study, we have attempted to evaluate if ingestion of certain spices can activate heat-generating mechanisms in the body such that a fall in. core body temperature (CBT) can be delayed or prevented when faced with a cold challenge. Ten commonly used spices in the Indian cuisine were chosen and 70% ethanol extract of the spices were fed orally to male Wistar rats at a dose of 250mg/kg for a period of 7 days. A change in CBT during cold exposure was recorded before and after treatment. At the end of the experiment, plasma norepinephrine and serum free fatty acid levels were estimated. We observed that among the ten spices, treatment with cinnamon and pepper extracts showed significant improvement in comparison to the control group. Based on evidence in literature and the HPLC-MS analysis from our lab, we hypothesized that the effects of the pepper and cinnamon extracts might be due to their piperine and cinnamaldehyde content respectively. However, no improved endurance was observed when they were administered alone. Poor endurance following depletion of endogenous norepinephrine levels using reserpine indicated its involvement in mediating the heat generating processes. However, it is noteworthy that green tea and spice treated animals exhibited a fall in CBT which was lower than their initial fall. In conclusion, our findings provide experimental evidence that ingestion of spices, viz., pepper and cinnamon, might elicit thermogenic responses such that hypothermia can be delayed or prevented upon cold exposure.

Plant regeneration through somatic embryogenesis and genome size analysis of Coriandrum sativum L.

In the present study, an improved plant regeneration protocol via primary and secondary somatic embryogenesis was established in two Co-1 and Rajendra Swathi (RS) varieties of Coriandrum sativum L. Callus was induced from root explants on 2, 4-D (0.5-2.0 mg/l) supplemented MS. The addition of BA (0.2 mg/l) improved callus induction and proliferation response significantly. The maximum callus induction frequency was on 1.0 mg/l 2, 4-D and 0.2 mg/l BA added MS medium (77.5 % in Co-1 and 72.3 % in RS). The callus transformed into embryogenic callus on 2, 4-D added MS with maximum embryogenic frequency was on 1.0 mg/l. The granular embryogenic callus differentiated into globular embryos on induction medium, which later progressed to heart-, torpedo- and cotyledonary embryos on medium amended with 0.5 mg/l NAA and 0.2 mg/l BA. On an average, 2-3 secondary somatic embryos (SEs) were developed on mature primary SEs, which increased the total embryo numbers in culture. Histology and scanning electron microscopy (SEM) studies are presented for the origin, development of primary and secondary embryos in coriander. Later, these induced embryos converted into plantlets on 1.0 mg/l BA and 0.2 mg/l NAA-amended medium. The regenerated plantlets were cultured on 0.5 mg/l IBA added ½ MS for promotion of roots. The well-rooted plantlets were acclimatized and transferred to soil. The genetic stability of embryo-regenerated plant was analyzed by flow cytometry with optimized Pongamia pinnata as standard. The 2C DNA content of RS coriander variety was estimated to 5.1 pg; the primary and secondary somatic embryo-derived plants had 5.26 and 5.44 pg 2C DNA content, respectively. The regenerated plants were genetically stable, genome size similar to seed-germinated coriander plants.

System-Wide Adaptations of Desulfovibrio alaskensis G20 to Phosphate-Limited Conditions.

The prevalence of lipids devoid of phosphorus suggests that the availability of phosphorus limits microbial growth and activity in many anoxic, stratified environments. To better understand the response of anaerobic bacteria to phosphate limitation and starvation, this study combines microscopic and lipid analyses with the measurements of fitness of pooled barcoded transposon mutants of the model sulfate reducing bacterium Desulfovibrio alaskensis G20. Phosphate-limited G20 has lower growth rates and replaces more than 90% of its membrane phospholipids by a mixture of monoglycosyl diacylglycerol (MGDG), glycuronic acid diacylglycerol (GADG) and ornithine lipids, lacks polyphosphate granules, and synthesizes other cellular inclusions. Analyses of pooled and individual mutants reveal the importance of the high-affinity phosphate transport system (the Pst system), PhoR, and glycolipid and ornithine lipid synthases during phosphate limitation. The phosphate-dependent synthesis of MGDG in G20 and the widespread occurrence of the MGDG/GADG synthase among sulfate reducing ∂-Proteobacteria implicate these microbes in the production of abundant MGDG in anaerobic environments where the concentrations of phosphate are lower than 10 µM. Numerous predicted changes in the composition of the cell envelope and systems involved in transport, maintenance of cytoplasmic redox potential, central metabolism and regulatory pathways also suggest an impact of phosphate limitation on the susceptibility of sulfate reducing bacteria to other anthropogenic or environmental stresses.

CyAbrB2 Contributes to the Transcriptional Regulation of Low CO2 Acclimation in Synechocystis sp. PCC 6803.

Acclimation to low CO2 conditions in cyanobacteria involves the co-ordinated regulation of genes mainly encoding components of the carbon-concentrating mechanism (CCM). Making use of several independent microarray data sets, a core set of CO2-regulated genes was defined for the model strain Synechocystis sp. PCC 6803. On the transcriptional level, the CCM is mainly regulated by the well-characterized transcriptional regulators NdhR (= CcmR) and CmpR. However, the role of an additional regulatory protein, namely cyAbrB2 belonging to the widely distributed AbrB regulator family that was originally characterized in the genus Bacillus, is less defined. Here we present results of transcriptomic and metabolic profiling of the wild type and a ΔcyabrB2 mutant of Synechocystis sp. PCC 6803 after shifts from high CO2 (5% in air, HC) to low CO2 (0.04%, LC). Evaluation of the transcriptomic data revealed that cyAbrB2 is involved in the regulation of several CCM-related genes such as sbtA/B, ndhF3/ndhD3/cupA and cmpABCD under LC conditions, but apparently acts supplementary to NdhR and CmpR. Under HC conditions, cyAbrB2 deletion affects the transcript abundance of PSII subunits, light-harvesting components and Calvin-Benson-Bassham cycle enzymes. These changes are also reflected by down-regulation of primary metabolite pools. The data suggest a role for cyAbrB2 in adjusting primary carbon and nitrogen metabolism to photosynthetic activity under fluctuating environmental conditions. The findings were integrated into the current knowledge about the acquisition of inorganic carbon (Ci), the CCM and parts of its regulation on the transcriptional level.

Chronic Hypobaric Hypoxia Induces Right Ventricular Hypertrophy and Apoptosis in Rats: Therapeutic Potential of Nanocurcumin in Improving Adaptation.

Nehra, Sarita, Varun Bhardwaj, Santosh Kar, and Deepika Saraswat. Chronic hypobaric hypoxia induces right ventricular hypertrophy and apoptosis in rats: therapeutic potential of nanocurcumin in improving adaptation. High Alt Med Biol. 17:342-352, 2016.-a sustained work load on the right heart on ascent to high altitudes promotes right ventricular hypertrophy (RVH), which eventually undergoes decompensation and promotes pathological damage. However, the exact set of events leading to damage remains unidentified. Curcumin is a natural antioxidant and antihypertrophic agent, but it has poor biostability. Nanotized curcumin (nanocurcumin) has emerged as a promising agent with improved biostability while retaining the therapeutic properties of curcumin. The present study aimed at analyzing the therapeutic properties of nanocurcumin in ameliorating cardiac damage due to chronic hypobaric hypoxia (HH)-induced RVH in comparison to curcumin. Sprague-Dawley rats exposed to HH (25,000 feet, effective oxygen fraction in air [FIO2] ∼0.08, temperature 28°C ± 1°C, relative humidity 55% ± 2% for 3, 7, 14, and 21 days) developed RVH with increased interstitial collagen content, Fulton's index, and cardiomyocyte cross-sectional area while upregulating atrial natriuretic peptide. Tissue damage due to apoptotic cell death was evident by cytochrome-c/caspase-3 activation and TUNEL assay. Concomitant modulation of cyclic guanosine monophosphate (cGMP)/cGK-1, calmodulin-dependent protein kinase II (CaMkinase II), and intracellular calcium levels with increased free radical-induced damage and lipid peroxidation further contributed to the right heart pathology. Nanocurcumin supplementation decreased HH-induced RVH and apoptosis while modulating cardiac cGMP/cGK-1 signaling, and maintaining CaMkinase II, intracellular calcium levels and redox status better than curcumin. Nanocurcumin-mediated antiapoptotic effects might have benefited residents and sojourners at high altitude in preventing hypoxic cardiac damage.

Biomimetic synthesis of antimicrobial silver nanoparticles using in vitro-propagated plantlets of a medicinally important endangered species: Phlomis bracteosa.

In vitro-derived cultures of plants offer a great potential for rapid biosynthesis of chemical-free antimicrobial silver nanoparticles (AgNPs) by enhancing their phytochemical reducing potential. Here, we developed an efficient protocol for in vitro micropropagation of a high-value endangered medicinal plant species, Phlomis bracteosa, in order to explore its biogenic potential in biomimetic synthesis of antimicrobial AgNPs. Murashige and Skoog medium supplemented with 2.0 mg/L thidiazuron was found to be more efficient in inducing optimum in vitro shoot regeneration (78%±4.09%), and 2.0 mg/L indole-3-butyric acid was used for maximum root induction (86%±4.457%). Antimicrobial AgNPs were successfully synthesized by using aqueous extract (rich in total phenolics and flavonoids content) of in vitro derived plantlets of P. bracteosa. Ultraviolet-visible spectroscopy of synthesized AgNPs showed characteristic surface plasmon band in the range of 420-429 nm. The crystallinity, size, and shape of the AgNPs were characterized by X-ray diffraction and scanning electron microscopy. Face-centered cubic AgNPs of almost uniform spherical size (22.41 nm) were synthesized within a short time (1 hour) at room temperature. Fourier-transform infrared spectroscopy revealed that the polyphenols were mainly responsible for reduction and capping of synthesized AgNPs. Energy dispersive X-ray analysis further endorsed the presence of elemental silver in synthesized AgNPs. These biosynthesized AgNPs displayed significantly higher bactericidal activity against multiple drug-resistant human pathogens. The present work highlighted the potent role of in vitro-derived plantlets of P. bracteosa for feasible biosynthesis of antimicrobial AgNPs, which can be used as nanomedicines in many biomedical applications.

Photorespiration participates in the assimilation of acetate in Chlorella sorokiniana under high light.

The development of microalgae on an industrial scale largely depends on the economic feasibility of mass production. High light induces productive suspensions during cultivation in a tubular photobioreactor. Herein, we report that high light, which inhibited the growth of Chlorella sorokiniana under autotrophic conditions, enhanced the growth of this alga in the presence of acetate. We compared pigments, proteomics and the metabolic flux ratio in C. sorokiniana cultivated under high light (HL) and under low light (LL) in the presence of acetate. Our results showed that high light induced the synthesis of xanthophyll and suppressed the synthesis of chlorophylls. Acetate in the medium was exhausted much more rapidly in HL than in LL. The data obtained from LC-MS/MS indicated that high light enhanced photorespiration, the Calvin cycle and the glyoxylate cycle of mixotrophic C. sorokiniana. The results of metabolic flux ratio analysis showed that the majority of the assimilated carbon derived from supplemented acetate, and photorespiratory glyoxylate could enter the glyoxylate cycle. Based on these data, we conclude that photorespiration provides glyoxylate to speed up the glyoxylate cycle, and releases acetate-derived CO2 for the Calvin cycle. Thus, photorespiration connects the glyoxylate cycle and the Calvin cycle, and participates in the assimilation of supplemented acetate in C. sorokiniana under high light.

Acute L-arginine supplementation has no effect on cardiovascular or thermoregulatory responses to rest, exercise, and recovery in the heat.

PURPOSE: To investigate the effect of acute L-arginine (L-ARG) supplementation on cardiovascular and thermoregulatory responses to rest, exercise, and recovery in the heat. METHODS: Eight healthy men (age 27 ± 6 years; stature 176 ± 6 cm; body mass 76 ± 4 kg; maximal power output 237 ± 39 W) participated in a double-blind, crossover study, attending the laboratory for two experimental trials. On each occasion, participants consumed 500 ml of a black currant-flavoured cordial beverage 30 min before completing a 90 min experiment in the heat (35 °C and 50% rh). The experiment consisted of 30 min of seated rest, followed by 30 min submaximal cycling (60% maximal power output) and 30 min passive seated recovery. On one visit the drink contained 10 g of dissolved L-ARG while on the other visit it did not. RESULTS: L-ARG supplementation increased plasma L-ARG concentrations (peak +223 ± 80% after 60 min of the 90 min experiment); however, supplementation had no effect on rectal temperature, mean skin temperature, heart rate, arterial pressure, forearm skin vascular conductance, oxygen consumption or sweat loss at rest, during exercise, or during recovery in the heat (p > 0.05). CONCLUSION: Acute ingestion of 10 g L-ARG supplementation failed to elicit any changes in the cardiovascular or thermoregulatory responses to active or passive heat exposure in young, healthy males.

The study of ascorbate peroxidase, catalase and peroxidase during in vitro regeneration of Argyrolobium roseum.

Here, we demonstrate the micropropagation protocol of Argyrolobium roseum (Camb.), an endangered herb exhibiting anti-diabetic and immune-suppressant properties, and antioxidant enzymes pattern is evaluated. Maximum callogenic response (60 %) was observed from leaf explant at 1.0 mg L(-1) 1-nephthalene acetic acid (NAA) and 0.5 mg L(-1) 6-benzyl aminopurine (BA) in Murashige and Skoog (MS) medium using hypocotyl and root explants (48 % each). Addition of AgNO3 and PVP in the culture medium led to an increase in callogenic response up to 86 % from leaf explant and 72 % from hypocotyl and root explants. The best shooting response was observed in the presence of NAA, while maximum shoot length and number of shoots were achieved based on BA-supplemented MS medium. The regenerated shoots were rooted and successfully acclimatized under greenhouse conditions. Catalase and peroxidase enzymes showed ascending pattern during in vitro plant development from seed while ascorbate peroxidase showed descending pattern. Totally reverse response of these enzymes was observed during callus induction from three different explants. During shoot induction, catalase and peroxidase increased at high rate while there was a mild reduction in ascorbate peroxidase activity. Catalase and peroxidase continuously increased; on the other hand, ascorbate peroxidase activity decreased during root development and acclimatization states. The protocol described here can be employed for the mass propagation and genetic transformation of this rare herb. This study also highlights the importance and role of ascorbate peroxidase, catalase, and peroxidase in the establishment of A. roseum in vitro culture through callogenesis and organogenesis.

Design and conduct of 'Xtreme Alps': a double-blind, randomised controlled study of the effects of dietary nitrate supplementation on acclimatisation to high altitude.

The study of healthy human volunteers ascending to high altitude provides a robust model of the complex physiological interplay that emulates human adaptation to hypoxaemia in clinical conditions. Nitric oxide (NO) metabolism may play an important role in both adaptation to high altitude and response to hypoxaemia during critical illness at sea level. Circulating nitrate and nitrite concentrations can be augmented by dietary supplementation and this is associated with improved exercise performance and mitochondrial efficiency. We hypothesised that the administration of a dietary substance (beetroot juice) rich in nitrate would improve oxygen efficiency during exercise at high altitude by enhancing tissue microcirculatory blood flow and oxygenation. Furthermore, nitrate supplementation would lead to measurable increases in NO bioactivity throughout the body. This methodological manuscript describes the design and conduct of the 'Xtreme Alps' expedition, a double-blind randomised controlled trial investigating the effects of dietary nitrate supplementation on acclimatisation to hypobaric hypoxia at high altitude in healthy human volunteers. The primary outcome measure was the change in oxygen efficiency during exercise at high altitude between participants allocated to receive nitrate supplementation and those receiving a placebo. A number of secondary measures were recorded, including exercise capacity, peripheral and microcirculatory blood flow and tissue oxygenation. Results from this study will further elucidate the role of NO in adaption to hypoxaemia and guide clinical trials in critically ill patients. Improved understanding of hypoxaemia in critical illness may provide new therapeutic avenues for interventions that will improve survival in critically ill patients.

Thyroid hormone regulates muscle function during cold acclimation in zebrafish (Danio rerio).

Thyroid hormone (TH) is a universal regulator of growth, development and metabolism during cold exposure in mammals. In zebrafish (Danio rerio), TH regulates locomotor performance and metabolism during cold acclimation. The influence of TH on locomotor performance may be via its effect on metabolism or, as has been shown in mammals, by modulating muscle phenotypes. Our aim was to determine whether TH influences muscle phenotypes in zebrafish, and whether this could explain changes in swimming capacity in response to thermal acclimation. We used propylthiouracil and iopanoic acid to induce hypothyroidism in zebrafish over a 3-week acclimation period to either 18 or 28°C. To verify that physiological changes following hypothyroid treatment were in fact due to the action of TH, we supplemented hypothyroid fish with 3,5-diiodothryronine (T2) or 3,5,3'-triiodothyronine (T3). Cold-acclimated fish had significantly greater sustained swimming performance (Ucrit) but not burst speed. Greater Ucrit was accompanied by increased tail beat frequency, but there was no change in tail beat amplitude. Hypothyroidism significantly decreased Ucrit and burst performance, as well as tail beat frequency and SERCA activity in cold-acclimated fish. However, myofibrillar ATPase activity increased in cold-acclimated hypothyroid fish. Hypothyroid treatment also decreased mRNA concentrations of myosin heavy chain fast isoforms and SERCA 1 isoform in cold-acclimated fish. SERCA 1 mRNA increased in warm-acclimated hypothyroid fish, and SERCA 3 mRNA decreased in both cold- and warm-acclimated hypothyroid fish. Supplementation with either T2 or T3 restored Ucrit, burst speed, tail beat frequency, SERCA activity and myosin heavy chain and SERCA 1 and 3 mRNA levels of hypothyroid fish back to control levels. We show that in addition to regulating development and metabolism in vertebrates, TH also regulates muscle physiology in ways that affect locomotor performance in fish. We suggest that the role of TH in modulating SERCA1 expression during cold exposure may have predisposed it to regulate endothermic thermogenesis.