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Most real-world systems are characterised by dynamics and correlations emerging at multiple time scales, and are therefore referred to as complex systems. In this work, the complexity of time series produced by complex systems was investigated in the frame of information theory computing the entropy rate via the conditional entropy (CE) measure. A comparative investigation of several CE estimators, based on linear parametric and non-linear model-free representations of the process dynamics, was performed considering simulated linear autoregressive (AR) and mixed non-linear deterministic and linear stochastic dynamics processes, as well as physiological time series reflecting short-term cardiorespiratory dynamics. In simulations, the estimated CE values decreased when reducing the system complexity through an increase in the pole radius of the AR process or with the predominance of the deterministic behaviour in the mixed dynamics. In the application to cardiorespiratory dynamics, a reduction in physiological complexity was observed resulting from a regularization of the time series of heart rate and respiratory volume when decreasing the breathing rate. Our results evidence how simple and fast approaches based on linear parametric or permutation-based model-free estimators allow efficient discrimination of complexity changes in the short-term evolution of complex dynamic systems. However, in the presence of non-linear dynamics, the superiority of the more general but computationally expensive nearest-neighbour method is highlighted. These findings have implications for the assessment of complex dynamics both in clinical settings and in physiological monitoring. [ABSTRACT FROM AUTHOR]

Recurrent ankle sprains are common in soccer players, characterized by restricted range of motion, pain, and decreased proprioception, strength, and postural control. The objective was to evaluate the effectiveness of a fascial therapy and strength training program, combined with kinesiotaping, in improving ankle range of motion, pain, strength and stability in footballers with recurrent sprains. A simple blind randomized clinical trial was conducted on soccer players. Thirty-six federated footballers were recruited and randomized to the two study groups. The experimental group received an intervention using myofascial techniques applied to the subastragaline joint, eccentric training with an isoinertial device and neuromuscular taping. The control group was administered an intervention using myofascial techniques on the subastragaline joint and eccentric training with an isoinertial device. The results were recorded for all players at baseline, after 4 weeks of intervention, and at the end of the 4-week follow-up period. Subsequent to intervention and follow-up, we found statistically significant improvements in the experimental group in ankle mobility, strength and stability. The control group exhibited improvements in all study variables. No differences in the improvement of variables were found based on the allocation of athletes to one group or another. The combination of fascial therapy and eccentric strength training with an isoinertial device improves ankle mobility, strength and stability in footballers with recurrent ankle sprains. The use of taping techniques failed to provide a greater improvement of the study variables when combined with manual therapy and strength techniques. • Recurrent ankle sprains are common in soccer players. • Myofascial therapy, strength training and kinesiotaping can improve pain and ankle mobility. • This protocol can improve ankle strength and stability of the lower limbs. • The improvement of ankle functionality regardless of the use of stabilizing taping. [ABSTRACT FROM AUTHOR]

Background and Aims: Novel genetic determinants associated with coronary artery disease (CAD) have been discovered by genome wide association studies. Variants encompassing the CELSR2- PSRC1-SORT1 gene cluster have been associated with CAD. This study is aimed to investigate the rs629301 polymorphism association with the extent of CAD evaluated by coronary angiography (CAG), and to evaluate its associations with an extensive panel of lipid and lipoprotein measurements in a large Italian cohort of 2429 patients.Methods and Results: The patients were collected by four Intensive Care Units located in Palermo and Verona (Italy). Clinical Records were filed, blood samples were collected, lipids and apolipoproteins (apo) were measured in separate laboratories. CAD was defined by the presence of stenotic arteries (>50% lumen diameter) by CAG. The presence of CAD was associated with the rs629301 genotype. Patients with CAD were 78% and 73% (p = 0.007) of the T/T vs. T/G + G/G genotype carriers respectively. T/T genotype was also correlated with the number of stenotic arteries, with a 1.29 (1.04-1.61) risk to have a three-arteries disease. T/T genotype correlated with higher levels of LDL-, non-HDL cholesterol, apoB, apoE and apoCIII, and lower HDL-cholesterol. Logistic Regression confirmed that rs629301was associated with CAD independently from the common risk factors, with a risk similar to that conferred by ten years of age [odds ratios were 1.43 (1.04-1.96) and 1.39 (1.22-1.58) respectively].Conclusions: rs629301 risk allele was independently associated with the extension and severity of CAD and positively with apoE and apoB containing lipoproteins. [ABSTRACT FROM AUTHOR]

Brain–heart interactions involve bidirectional effects produced by bottom-up input at each heartbeat, and top-down neural regulatory responses of the brain. While the cortical processing of the heartbeat is usually investigated through the analysis of the Heartbeat Evoked Potential, in this study we propose an alternative approach based on the variability in the predictability of the brain dynamics induced by the heartbeat. In a group of eighteen subjects in whom simultaneous recording of the electroencephalogram (EEG) and electrocardiogram was performed in a resting-state, we analyzed the temporal profile of the local Information Storage (IS) to detect changes in the regularity of EEG signals in time windows associated with different phases of the cardiac cycle at rest. The average values of the local IS were significantly higher in the parieto-occipital areas of the scalp, suggesting an activation of the Default Mode Network, regardless of the cardiac cycle phase. In contrast, the variability of the local IS showed marked differences across the cardiac cycle phases. Our results suggest that cardiac activity influences the predictive information of EEG dynamics differently in the various phases of the cardiac cycle. The variability of local IS measures can represent a useful index to identify spatio-temporal dynamics within the neurocardiac system, which generally remain overlooked by the more widely employed global measures. • An alternative method to explore cortical heartbeat processing is proposed. • Pointwise time-specific measures are used to characterize brain–heart interactions. • Local information storage measures reveal dynamics hidden globally. • Cardiac timing affects changes in the regularity of brain activity. [ABSTRACT FROM AUTHOR]

Brain–heart interactions involve bidirectional effects produced by bottom-up input at each heartbeat, and top-down neural regulatory responses of the brain. While the cortical processing of the heartbeat is usually investigated through the analysis of the Heartbeat Evoked Potential, in this study we propose an alternative approach based on the variability in the predictability of the brain dynamics induced by the heartbeat. In a group of eighteen subjects in whom simultaneous recording of the electroencephalogram (EEG) and electrocardiogram was performed in a resting-state, we analyzed the temporal profile of the local Information Storage (IS) to detect changes in the regularity of EEG signals in time windows associated with different phases of the cardiac cycle at rest. The average values of the local IS were significantly higher in the parieto-occipital areas of the scalp, suggesting an activation of the Default Mode Network, regardless of the cardiac cycle phase. In contrast, the variability of the local IS showed marked differences across the cardiac cycle phases. Our results suggest that cardiac activity influences the predictive information of EEG dynamics differently in the various phases of the cardiac cycle. The variability of local IS measures can represent a useful index to identify spatio-temporal dynamics within the neurocardiac system, which generally remain overlooked by the more widely employed global measures. • An alternative method to explore cortical heartbeat processing is proposed. • Pointwise time-specific measures are used to characterize brain–heart interactions. • Local information storage measures reveal dynamics hidden globally. • Cardiac timing affects changes in the regularity of brain activity. [ABSTRACT FROM AUTHOR]

Metabolomics is a rapidly emerging discipline within functional genomics which is increasingly being applied to understand biochemical phenotypes across a range of biological systems. Metabolomics measures all (or a subset) metabolites in a cell at a specific time point, reflecting a snapshot of all the regulatory events responding to the external environmental conditions. Although metabolomics and systems biology approaches have been applied to the study of terrestrial plants, few marine macrophytes have been examined using these novel technologies. Marine macrophytes (including seaweeds and seagrasses) are marine ecosystem engineers delivering a range of ecologically and economically valuable biological services; however they are under threat from a wide range of anthropogenic stressors, climate variation, invasive species and pathogens. Investigating metabolomic regulation in these organisms is crucial to understand their acclimation, adaptation and defence responses to environmental challenges. This review describes the current analytical tools available to study metabolomics in marine macrophytes, along with their limitations for both targeted and non-targeted workflows. To illustrate recent advances in systems biology studies in marine macrophytes, we describe how metabolites are used in chemical defence to deter a broad range of invasive species and pathogens, as well as metabolomic reprogramming leading to acclimation or adaptive strategies to environmental and anthropogenic stresses. Where possible, the mechanistic processes associated with primary and secondary plant metabolism governing cellular homeostasis under extreme environments are discussed. Further, we provide a comprehensive overview of an in silico plant metabolome database that can be utilized to advance our knowledge from a system biology approach to marine macrophytes. Finally, functional integration of metabolomics with the allied “omics” disciplines of transcriptomics and proteomics, as well as the emerging discipline of “fluxomics” are discussed in the context of developing biological system networks, the identification of unknown gene/protein functions and the analysis of metabolic pathways in marine plants exposed to stress. [ABSTRACT FROM AUTHOR]

Hypoxia (low oxygen stress) is increasingly reported on coral reefs, caused by ocean deoxygenation linked to coastal nutrient pollution and ocean warming. While the ability to regulate respiration is a key driver of hypoxia tolerance in many other aquatic taxa, corals' oxyregulatory capabilities remain virtually unexplored. Here, we examine O 2 -consumption patterns across 17 coral species under declining O 2 partial pressure (p O 2). All corals showed ability to oxyregulate, but total positive regulation (T pos) varied between species, ranging from 0.41 (Pocillopora damicornis) to 2.42 (P. acuta). On average, corals performed maximum regulation effort (P cmax) at low p O 2 (30% air saturation, corresponding to lower O 2 levels measured on natural reef systems), and exhibited detectable regulation down to as low as <10% air saturation. Our study shows that corals are not oxyconformers as previously thought, suggesting oxyregulation is likely important for survival in dynamic O 2 environments of shallow coral reefs subjected to hypoxic events. • Respiratory responses of reef building corals were assessed under progressive hypoxia. • Corals are capable of moderate oxyregulation despite their anatomical simplicity. • Significant variability in positive regulation exists among reef-building corals. • Oxyregulation occurs down to very low PO 2 values (30% air saturation). [ABSTRACT FROM AUTHOR]

Poly(3-hydroxybutyrate) (PHB) is a promising bioplastic compound which is produced by certain prokaryotic microbes such as some species of photosynthetic cyanobacteria. The production of cyanobacterial PHB has the potential to reduce traditional PHB production costs through using atmospheric CO 2 as a carbon substrate, as opposed to using expensive organic carbon substrates needed for heterotrophic PHB production. This techno-economic analysis explores the financial profitability of using open raceway ponds to cultivate the cyanobacteria and solvent extraction on biomass to produce 10,000 tonnes of PHB per year. With a 10% PHB yield (10% PHB mass fraction of cell dry weight) used, the capital cost for the plant was $193.5 M USD and the operating cost was $147.3 M USD per year. At $4000 per tonne for PHB, the revenue of this plant was only $40 M USD per year, resulting in a 20-year Net Present Value (NPV) of -$1.8B USD. The breakeven minimum PHB selling price (for a zero 20-year NPV) was $18,339 USD per tonne. From the sensitivity analysis it was revealed that the key driver behind profitability of cyanobacterial PHB production is PHB yield. Lastly, seven different hypothetical scenarios are explored to increase the economic viability of the cyanobacterial production. These strategies ranged from additional revenue streams from pigment extraction, treatment of wastewater, to decreasing capital costs by using deep 'PHB ripening ponds'. Operational costs were reduced through use of flocculants, solar power purchase agreements and an on-site anaerobic digester to reduce both energy and nutrient costs through digestate use as pond media. The optimistic combination of all these factors resulted in a final minimum PHB selling price of $7704 USD per tonne; still roughly twice the current market value of PHB. [Display omitted] • Assesses the economic viability of an industrial scale cyanobacterial PHB bioplastic production facility. • Identifies the key capital and operating cost barriers through techno-economic modelling and sensitivity analysis. • Quantifies the minimum cyanobacterial PHB selling price to achieve financial viability and compares to current market prices. • Explores several strategies to improve profitability of cyanobacterial PHB production through scenario analysis (including a biorefinery approach and incorporation of wastewater treatment). [ABSTRACT FROM AUTHOR]

Lidocaine is commonly used for topical anesthesia of the upper airway in patient with anticipated difficult tracheal intubation undergoing awake fiberoptic intubation. Lidocaine toxicity is dose related and proportional to its plasma level. Although neurologic toxicity has been frequently observed with intravenous use, it has also been reported for topical use. We report on a case of a patient with base tongue abscess who developed sudden seizures and coma during application of topical anesthesia with lidocaine for awake fiberoptic intubation. The presence of a deep neck infection that causes hyperemia and edema of the pharyngolaryngeal mucosa may enhance transmucosal systemic absorption of local anesthetic. Moreover, conditions such as hypercarbia, dysphagia, or hepatic diseases are known to facilitate onset of lidocaine neurologic toxicity with serum concentration lower than normal. These findings should be kept in mind before administering topical anesthesia of the upper airway. In the presence of any of these conditions above, either the total dose of local anesthetic or its concentration should be reduced as much as possible. [ABSTRACT FROM AUTHOR]

Objective: The Jarvik 2000 (Jarvik Heart, Inc, New York, NY) is a thumb-sized high-speed impeller pump that is used as a ventricular assist device in patients with terminal heart failure. Because the Jarvik 2000 is designed for long-term use, it is a central question whether the mechanical forces inside the pump affect blood components. This study evaluated the potential association of the high rotational speed of the Jarvik 2000 with platelet activation, which may result in thromboembolic events. Methods: The study group comprised patients with terminal heart failure who were supported with the Jarvik 2000. All were men and received 100 mg aspirin daily. In 8 patients, soluble platelet activation markers (P-selectin and sCD40L), platelet counts, and hemolysis markers (haptoglobin and lactate dehydrogenase levels) were determined. In 5 patients, P-selectin expression and platelet receptor glycoprotein IIb/IIIa activation were determined with flow cytometry and compared with a control group of 5 healthy men. Platelet activation was measured at various rotational device speeds. Results: After Jarvik 2000 implantation, increased hemolysis was observed, but platelet activation markers and platelet counts were not affected. Increased rotational speed (8000 to 12,000 rpm) of the device also did not result in increased platelet activation. Conclusion: The Jarvik 2000 was not associated with detectable platelet activation, despite high rotational impeller speeds. [Copyright &y& Elsevier]

Abstract: Triglycerides (TGs) are vehicled by multiple particles with different abilities to promote atherosclerosis. Among plasma TG-rich lipoproteins (TRLs), subspecies may or may not contain apolipoprotein E (apoE) molecules: in this study, we evaluated the relative contribution of apoE-rich and apoE-poor TRLs to coronary atherosclerosis. We selected a group of males with premature coronary artery disease (CAD) without any of the classical nonlipid risk factors and/or high plasma lipid levels and evaluated the plasma concentration of TRL subspecies in comparison with healthy controls. Patients with CAD and controls had total cholesterol and TG levels within the normal range (despite slightly, even if significantly, higher TG levels in patients with CAD) and low-density lipoprotein cholesterol levels near optimal values. Nevertheless, patients with CAD had significantly lower high-density lipoprotein cholesterol, smaller low-density lipoprotein peak particle size, and a reduced HDL2b subfraction than controls. In addition, we observed higher concentrations of total TRL in patients with CAD together with a selective increase in apoE-rich particles. All these data were confirmed after correction for TG levels. We also investigated which parameters were associated with the spread of coronary atherosclerosis. Subjects with a single-vessel disease had selectively lower levels of apoE-rich fractions than patients with a multivessel disease. This was confirmed by multivariate analysis. Patients with a premature CAD free of nonlipid conventional risk factors, despite not having elevated lipid levels, show several lipoprotein abnormalities. Besides known atherogenic alterations, the accumulation of apoE-rich TRL subfractions may represent an additive factor that can potentially promote and initiate the atherosclerotic process. [Copyright &y& Elsevier]

Summary: Background and aim: In this study we addressed some open questions in patients with coronary artery disease (CAD). First, we analysed which of the traditional risk factors was associated with the spreading of coronary stenosis and second, we aimed to identify if any variable was predictive of post-percutaneous transluminal coronary angioplasty (PTCA) clinical events. Methods and results: We collected a consecutive series of patients with CAD (n =301) and in the subgroup of patients undergoing PTCA (n =135) we performed a prospective one-year follow-up study recording cardiovascular morbidity and total mortality. According to the extension of coronary atherosclerosis, we found a significant relationship with the prevalence of diabetes in men and with plasma HDL-cholesterol concentrations in women. The follow-up was completed in 95% of patients; we did not document any death whereas clinical events were registered in 16% of patients. At univariate analysis, we found that patients with clinical events had a higher prevalence of family history of CAD (43% vs 14%, p <0.005), diabetes (52% vs 21%, p <0.005) and multivessel disease (52% vs 35%, p <0.05). Multivariate analysis (logistic regression) confirmed that family history of CAD (OR 4.6, 95% CI 1.7–12.8, p <0.005), diabetes (OR 4.0, 95% CI 1.5–10.6, p <0.01) and multivessel disease (OR 2.8, 95% CI 1.1–7.4, p <0.05) were the only variables predictive of clinical events. Conclusions: In this study, factors associated with the spreading of coronary stenosis were different according to the gender. Moreover, the presence of diabetes and multivessel disease had a negative impact on the long-term prognosis of patients undergoing PTCA. In addition, the family history of CAD represented in our study a strong predictor of clinical events. We suggest that in the management of post-PTCA patients, the role of individual baseline clinical characteristics must be taken into account and that subjects with a family history of premature CAD, diabetes and a wide extension of coronary disease represent those with the highest risk. [Copyright &y& Elsevier]

• Parametric approaches to assess spontaneous cardiovascular variability are studied. • A new causal local spectral decomposition method is used for baroreflex analysis. • Causal approaches allow to differentiate baroreflex and feedforward interactions. • Local approaches avoid confounding oscillatory effects outside the band of interest. • Causal local measures allow an objective assessment of baroreflex coupling and gain. The objective of this study is to present a new method to assess in the frequency domain the directed interactions between the spontaneous variability of systolic arterial pressure (SAP) and heart period (HP) from their linear model representation, and to apply it for studying the baroreflex control of arterial pressure in healthy physiological states and after acute myocardial infarction (AMI). The method is based on pole decomposition of the model transfer function and on the following evaluation of causal measures of coupling and gain from the poles associated to low frequency (0.04−0.15 Hz) oscillatory components. It is compared with traditional non-causal approaches for the spectral analysis of the baroreflex gain, and with causal approaches based on the directed coherence, in a group of AMI patients and in Young and Old healthy controls studied at rest and during head-up tilt. Analysis of feedforward interactions from RR to SAP is also performed. Our results support the importance of using local causal approaches to quantify separately baroreflex and feedforward interactions between RR and SAP, allowing both to confirm known physiological behaviors (e.g., weaker baroreflex effectiveness and lower sensitivity after AMI) and to suggest novel findings (e.g., preserved low frequency baroreflex response to postural stress after AMI). We also find that the postural stress alters feedforward interactions selectively across groups, being related to decreased coupling only in Young and to increased gain mostly in AMI. These results have a significance for the clinical assessment of the baroreflex and the physiological evaluation of cardiovascular interactions. [ABSTRACT FROM AUTHOR]

Phaeodactylum tricornutum is a model pennate diatom for molecular studies due to its sequenced genome, genetic tractability, and capacity to produce a myriad of compounds of biotechnological interest. While tools for genetic engineering and heterologous gene expression have been developed for this species, the available set of characterised promoters remains limited. A diverse and well-understood set of genetic regulatory elements is necessary to enable fine tuning of gene expression for metabolic engineering and synthetic biology approaches. In this study, we investigated the expression profile of four endogenous constitutive promoter and terminator pairs Nub (Phatr3_J14260), SVP (Phatr3_J16798), 45582 (Phatr3_J45582), and Pbt (Phatr3_J54958) selected from a multi-experiment, integrated transcriptomics database under a variety of growth conditions, including various light regimes and nitrogen (N) limitation. Expression of the fluorescent reporter mVenus was driven from non-integrative episomes and monitored via flow cytometry. Gene expression was validated on a transcript and protein level using RT-qPCR and western blot, respectively. We found that all four promoter and terminator pairs exhibited stable expression throughout exponential growth, with distinct expression patterns and reduced light dependency compared to the widely-used FcpB promoter. Furthermore, the activity of these promoters was less impacted by N depletion than for FcpB. Our results indicate these novel promoters and terminators are promising tools for fine tuning single or multiple gene expression in P. tricornutum. • Transcriptomics-guided selection of putative promoters/terminators in P. tricornutum. • Episomal expression of mVenus demonstrates promoters are suitable for genetic engineering. • Promoters drive extended expression during dark periods and nitrogen starvation. • Novel genetic parts are compatible with standardized TypeIIS cloning systems. • Clone-to-clone variation depends on promoter strength. [ABSTRACT FROM AUTHOR]

Microalgae are increasingly being considered for recombinant protein production because of low cultivation costs, absence of endotoxins and insusceptibility to human infectious agents. Despite these advantages, the yield of recombinant protein produced in microalgae is still low compared to more established expression systems and optimization at the genetic and cultivation levels is required for this new system to be economically viable. This study investigates the effect of biphasic temperature regimes on the yield of recombinant human interferon alpha 2a (IFN-α2a), a therapeutic protein known for its anti-cancer and anti-viral properties, produced by the model green alga Chlamydomonas reinhardtii (Cr.IFN-α2a). Biphasic growth is commonly employed to increase recombinant protein production in mammalian cell lines used for commercial production of therapeutic proteins, with a lowering of the temperature resulting in higher yields. In this study, lowering the temperature from 25 °C to 15 °C in mid-exponential growth phase increased the accumulation of Cr.IFN-α2a by 3.3-fold while it slowed down the growth of the three C. reinhardtii transgenic lines tested. In contrast, a rise of temperature from 25 °C to 35 °C accelerated cell growth, while negatively impacting the production of Cr.IFN-α2a. After a two-step chromatography purification, the Cr.IFN-α2a produced was estimated to be 53% pure with a yield of 90 μg/L of culture. The amino acid sequence of Cr.IFN-α2a was confirmed by mass spectrometry. However, the anti-viral activity of Cr.IFN-α2a was found to be 10 times lower than the human IFN-α2a standard produced using E. coli when challenged in a cytopathic effect (CPE) assay, likely due to the formation of aggregates. While the molecular mechanisms driving the accumulation of Cr.IFN-α2a at lower temperature remains unclear, our results support that reducing the temperature at the peak of expression is a valid strategy to increase the yield of recombinant Cr.IFN-α2a in C. reinhardtii. • First report of biphasic temperature regime on transgenic C. reinhardtii • A switch of temperature from 25 °C to 15 °C slowed down the growth of C. reinhardtii. • Lowering the temperature to 15 °C increased the yield of Cr.IFN-α2a by 3.3-fold. • The Cr.IFN-α2a was ~53% pure with a yield of 90 μg/L of culture after purification. • The Cr.IFN-α2a was less active than the human IFN-α2a standard produced in E. coli. [ABSTRACT FROM AUTHOR]

• Advances in cyanobacterial PHB metabolism hold potential for bioplastic production. • Key industrial strategies for efficient scale up are discussed. • Assessment of current economic status and key financial drivers identified. • Perspectives on future research areas to achieve economic viability. PHB is a promising bioplastic material that naturally accumulates in many strains of cyanobacteria. This comprehensive review covers recent advances in several topics including PHB metabolism, material properties, relevant extraction methods and protocols, industrial cultivation strategy, current economic assessment and much more. Ultimately, the profitability of cyanobacterial PHB production is controlled by low PHB productivity as well as expensive cultivation and harvesting equipment. Several research areas for improving viability of cyanobacterial PHB production have also been summarised and perspectives on future efforts suggested including; screening, genetic modification, wastewater cultivation and using chemical modulators among others. [ABSTRACT FROM AUTHOR]

Copper (Cu) in an essential trace metal but it can also contaminate coastal waters at high concentrations mainly from agricultural run-off and mining activities which are detrimental to marine organisms including seagrasses. The molecular mechanisms driving Cu toxicity in seagrasses are not clearly understood yet. Here, we investigated the molecular responses of the Australian seagrass, Z. muelleri at the whole transcriptomic level after 7 days of exposure to 250 μg Cu L−1 and 500 μg Cu L−1. The leaf-specific whole transcriptome results showed a concentration-dependent disturbance in chloroplast function, regulatory stress responses and defense mechanisms. This study provided new insights into the responses of seagrasses to trace metal stress and reports possible candidate genes which can be considered as biomarkers to improve conservation and management of seagrass meadows. • Copper toxicity in Z. muelleri is found to be concentration-dependent. • Exposure to Cu stress indicated disturbance in chloroplast function but no significant impact on photosystem subunits. • CytP450 (89A2-like) is found as copper-induced chemical defence mechanism. • Peroxidase and Glutathione s transferase are found as possible copper-induced enzymatic defence mechanisms at 500 μg Cu L-1 only. • Phytohormones and heavy metal-associated isoprenylated plant protein 3 (HIPP3) are activated as regulatory Cu stress responses. [ABSTRACT FROM AUTHOR]

• Three novel promoters were characterised in N. gaditana for transgene expression. • Hsp90 and EPPSII promoters outperformed TUB , a commonly used constitutive promoter. • These promoters expand N. gaditana genetic resource for basic and applied research. Nannochloropsis is a marine microalga from the Eustigmatophyceae stramenopile lineage that has been studied extensively due to a broad range of industrial applications, mostly related to their oil and pigment production. However, tools to genetically engineer members of this group, and therefore further understand and maximise their industrial potential are still limited. In order to expand the potential industrial uses of this organism, several molecular tools, including gene promoters of different strength, are needed. A comprehensive and diverse set of well-characterized promoters is key to a number of genetic engineering and synthetic biology applications, such as the assembly of complex biological functions or entire metabolic pathways. In this study, we measured the promoter activity of three endogenous constitutive promoters from N. gaditana genes EPPSII (Nga02101); HSP90 (Nga00934); ATPase (Nga06354.1) in driving the expression of a Sh ble- mVenus fluorescent reporter fusion protein. Through a combined approach that includes flow cytometry, RT-qPCR and immunoblotting, we profiled the activity of these promoters at both the transcript and protein level. Two promoters HSP90 (Nga00934) and EPPSII (Nga02101) outperformed the widely used β-tubulin promoter, exhibiting 4.5 and 3.1-fold higher mVenus fluorescence, respectively. A third promoter ATPase (Nga06354.1) was also able to drive the expression of transgenes, albeit at lower levels. We show that the new promoters identified in this study are valuable tools, which can be used for genetic engineering and functional genetics studies in N. gaditana. [ABSTRACT FROM AUTHOR]

Abstract Terpenoids are a large and diverse class of naturally occurring metabolites serving many industrial applications and natural roles. Economically important terpenoids are often produced in low abundance from their natural sources, making their industrial-scale production challenging or uneconomical, therefore engineered microorganisms are frequently used as heterologous production platforms. Photosynthetic microorganisms, such as the green alga Chlamydomonas reinhardtii , represent promising systems to produce terpenoids in a cost-effective and sustainable manner, but knowledge about the regulation of their terpenoid metabolism remains limited. Here we report on the investigation of the phytohormone methyl jasmonate (MeJA) as elicitor of algal terpenoid synthesis. We treated C. reinhardtii cells in mid-exponential growth phase with three different concentrations of MeJA (0.05, 0.5 and 1 mM). The highest concentration of MeJA affected the photosynthetic activity of the cells, arrested the growth and up-regulated key genes of the 2- C -methyl-D-erythritol 4-phosphate (MEP) pathway, leading to a significant increase in intermediates of this pathway, squalene and (S)-2,3-epoxysqualene, while the abundance of cycloartenol, and two main sterols (ergosterol and 7-dehydroporiferasterol) decreased. These data suggest the redirection of the carbon flux towards the synthesis of yet uncharacterised triterpenoid secondary metabolites upon MeJA treatment. Our results offer important new insights into the regulation of the triterpenoid metabolism in C. reinhardtii and raise important questions on hormonal signalling in microalgae. Phytohormone treatment is tested for the first time in algae, where it holds great potential for identifying key transcriptional regulators of the MEP pathway as targets for future metabolic engineering studies for improve production of high-value triterpenoids. Highlights • First report of triterpenoid elicitation by a phytohormone in C. reinhardtii. • MeJA treatment (1 mM) arrested the growth and up-regulated genes of the MEP pathway. • Highlighting a specialised pathway branching immediately upstream of cycloartenol. • Accumulation of an uncharacterised triterpenoid secondary metabolite (C 30 H 52 O 3). • Plant-like MeJA response machinery is putatively absent in C. reinhardtii. [ABSTRACT FROM AUTHOR]

Management of cervicocarcinoma during pregnancy is influenced by gestational age, stage of disease, and patient’s desire to maintain her pregnancy. We report a case of a pregnant patient with locally advanced cervicocarcinoma successfully treated by neoadjuvant chemotherapy, followed by caesarean section and radical surgery. [Copyright &y& Elsevier]