How to improve the energy-saving performance of China's transport sector? An input-output perspective.

The transport sector proves a major energy consumer in China, but improving energy-saving performance in China's provincial transport sector from the lifecycle perspective remains unresolved. Thus, this study employs the environmentally extended multi-region input-output (MRIO) method, structural path analysis, and the newest MRIO table of China from 2017, to investigate how to improve the energy-saving performance from final demand structure, supply chain, and pathway perspectives. The relevant results are threefold. (1) Regarding the final demand structure level, the embodied energy consumption of China's transport sector is predominantly driven by investment from the production side, while that of the consumption side is primarily caused by exports. (2) At the supply chain level, production-side embodied energy consumption primarily occurs along a three-echelon supply chain, while that from the consumption side mostly occurs via a two-echelon supply chain. (3) At the pathway level, the production-side energy-saving performance of China's provincial transport sector is dominated by two pathways along the construction sector, including transport sector → construction sector → final demands, and transport sector → intermediate inputs → construction sector → final demands, while that of the consumption side is chiefly determined by three pathways along internal transportation chains.

Continuous In Vivo Monitoring of Indole-3-Acetic Acid and Salicylic Acid in Tomato Leaf Veins Based on an Electrochemical Microsensor.

Indole-3-acetic acid (IAA) and salicylic acid (SA), as critical plant hormones, are involved in multiple physiological regulatory processes of plants. Simultaneous and continuous in vivo detection of IAA and SA will help clarify the mechanisms of their regulation and crosstalk. First, this study reports the development and application of an electrochemical microsensor for simultaneous and continuous in vivo detection of IAA and SA. This electrochemical microsensor system consisted of a tip (length, 2 mm) of platinum wire (diameter, 0.1 mm) modified with carbon cement and multi-walled carbon nanotubes, an untreated tip (length, 2 mm) of platinum wire (diameter, 0.1 mm), as well as a tip (length, 2 mm) of Ag/AgCl wire (diameter, 0.1 mm). It was capable of detecting IAA in the level ranging from 0.1 to 30 µM and SA ranging from 0.1 to 50 µM based on the differential pulse voltammetry or amperometric i-t., respectively. The dynamics of IAA and SA levels in tomato leaf veins under high salinity stress were continuously detected in vivo, and very little damage occurred. Compared to conventional detection methods, the constructed microsensor is not only suitable for continuously detecting IAA and SA in microscopic plant tissue in vivo, it also reduces the damage done to plants during the detection. More importantly, the continuous and dynamic changes in IAA and SA data obtained in stiu through this system not only can help clarify the interaction mechanisms of IAA and SA in plants, it also helps to evaluate the health status of plants, which will promote the development of basic research in botany and precision agriculture.

Protein-Directed Au(0)-Rich Gold Nanoclusters as Ratiometric Luminescence Sensors for Auric Ions via Comproportionation-Induced Emission Enhancement.

Gold nanoclusters (Au NCs) are widely used as fluorescent probes in biomedical sensing and imaging due to their versatile optical properties and low cytotoxicity. Surface engineering of gold nanoclusters (Au NCs) aims to design a surface with versatile physicochemical performances, but previous investigations have primarily focused on the acquisition of the "brightest" species. This has resulted in other types of Au NC being neglected. In the present study, our group prepared a series of Au NCs that were rich in surface Au(0), using the "aged" form of bovine serum albumin (BSA) via controlling the pH during synthesis. We found that slight increases of alkalinity during synthesis over that which produced Au NCs with the most intensive photoluminescence generated the "darkest" Au NCs, which exhibited the strongest absorption. These Au NCs included more Au atoms and had a higher Au(0) content. Furthermore, the addition of Au3+ quenched the emission of the "brightest" Au NCs, but increased that of the "darkest" Au NCs. The increased Au(I) proportion observed in the Au3+-treated "darkest" Au NCs resulted in a novel comproportionation-induced emission enhancement effect, which we utilized to construct a "turn-on" ratiometric sensor for toxic Au3+. The addition of Au3+ generated simultaneous, opposite effects on blue-emissive diTyr BSA residues and red-emissive Au NCs. After optimization, we successfully constructed ratiometric sensors for Au3+ with high sensitivity, selectivity, and accuracy. This study will inspire a new pathway to redesign the protein-framed Au NCs and analytical methodology via comproportionation chemistry.

Levilactobacillus yiduensis sp. nov., Isolated from Zha-chili in Yidu County, Hubei Province, China.

The zha-chili isolate HBUAS62285T is a facultative anaerobic strain. This bacterium was gram-positive, but it could not generate catalase, was not motile, did not form spores, had no flagella, and produced gamma-aminobutyric acid (GABA). Comparing of HBUAS62285T with its related type strains Levilactobacillus suantsaiihabitans BCRC 81129T, Levilactobacillus angrenensis M1530-1T, Levilactobacillus cerevisiae DSM 100836T, Levilactobacillus wangkuiensis 6-5(1)T, Levilactobacillus lanxiensis 13B17T, and Levilactobacillus mulengensis 112-3T showed that the similarity of 16S rRNA gene sequence was less than 99.13%. Strain HBUAS62285T has a G+C content of 50.57 mol%, an ANI value of less than 86.61%, an AAI value of less than 92.9%, and a dDDH value of less than 32.9% when compared to the aforementioned closely related strains. In the end, the most major fatty acids in cells were determined to be C16:0, C18:1 ω9c, C19:1 cyclo ω9,10c, and summed feature 10. Overall, the results of the phenotypic, genomic, chemotaxonomic, and phylogenetic studies indicate that strains HBUAS62285T and CD0817 constitute a new species within the genus Levilactobacillus, and the name Levilactobacillus yiduensis sp. nov. is proposed. The type strain is HBUAS62285T (= JCM 35804T = GDMCC 1.3507T).

How pom cheerleading improves the executive function of preschool children: the mediating role of speed and agility.

Physical exercises can improve individuals' physical health and cognition, but the internal influence path is unclear. This study aims to examine the influence of pom cheerleading training on physical fitness and executive function of preschool children and explore the relationship between sports training, physical fitness, and executive function. We selected seventy-one preschool children and divided them into the experimental group (n = 36) and the control group (n = 35). The experimental group kept a 12-week pom cheerleading training, and the exercises of the control group remained normal. Children's physical fitness and executive function were tested, in one week before and after the experiment, respectively. Results of repeated measurements analysis of variance and structural equation model test showed: (1) after 12-week pom cheerleading training, in terms of physical fitness, the experimental group has a significant improvement over the control group on agility and speed; in terms of executive function, the inhibitory control and working memory of the experimental group were significantly enhanced over the control group. (2) Speed quality plays a partial mediating role between pom cheerleading training and inhibitory control; agility plays a major mediating role between pom cheerleading training and working memory. It is concluded that physical exercise can directly improve preschool children's executive function, and indirectly enhance executive function mediated by physical fitness. Furthermore, structured and systematic physical education should be adopted for preschool children to cultivate their interest in sports and enhance their cognition.

Characterization of two novel pentose-fermenting and GABA-producing species: Levilactobacillus tujiorum sp. nov. and Secundilactobacillus angelensis sp. nov. Isolated from a solid-state fermented zha-chili.

Lactobacilli are dominant in zha-chili. This study provides a taxonomic characterization of five bacterial strains isolated from zha-chili in China. The cells were Gram-positive, facultative anaerobic, non-spore-forming, flagella-free, catalase-negative, heterofermentative, pentose-fermenting, and gamma-aminobutyric acid (GABA)-producing rods. For HBUAS51241T, HBUAS51329, and HBUAS51416, C16:0, C18:1ω9c and C19:0 iso were the predominant cellular fatty acids; diphosphatidylglycerol (DPG), phosphatidylglycerol (DP), glycolipids (GL), and glycolipids (AL) were the major phospholipids. While for HBUAS51383T and HBUAS58055, C16:0, C18:1ω9c, C19:0 cyclo ω8c were the predominant cellular fatty acids; DPG, DP, GL, and AL were the major phospholipids. Strains HBUAS51241T, HBUAS51329, and HBUAS51416 showed 98.1-99.1% 16S rRNA gene sequence similarity, 80.2-81.4% ANI, 87.7-90.0% AAI, and 23.8-32.8% digital DDH to their closest related type strains Levilactobacillus hammesii DSM 16381T, Levilactobacillus parabrevis ATCC 53295T, and Levilactobacillus fuyuanensis 244-4T. Strains HBUAS51383T and HBUAS58055 showed 98.7-99.5% 16S rRNA gene sequence similarity, 75.4-81.4% ANI, 75.5-89.1% AAI, and 19.7-24.0% digital DDH to their closest related type strains Secundilactobacillus silagincola IWT5T, Secundilactobacillus silagei JCM 19001T, Secundilactobacillus pentosiphilus IWT25T, Secundilactobacillus mixtipabuli IWT30T, Secundilactobacillus odoratitofui DSM 19909T, and Secundilactobacillus similis DSM 23365T. The central carbon metabolism pathways for the five strains were summarizeded. Based on the phenotypic, chemotaxonomic, and genomic data, we propose two novel species Levilactobacillus tujiorum sp. nov. whose type strain is HBUAS51241T (=GDMCC 1.3022T = JCM 35241T), and Secundilactobacillus angelensis sp. nov. whose type strain is HBUAS51383T (=GDMCC 1.3021T = JCM 35209T).

Tumor treating fields combined with a poly (adenosine diphosphate-ribose) polymerase inhibitor during radiotherapy for rapidly progressing IDH-wildtype diffuse astrocytoma: a case report.

A 57-year-old woman was diagnosed with IDH-wildtype (IDHwt) astrocytoma (World Health Organization grade II) with the molecular characteristics of glioblastoma. She underwent concurrent radiotherapy and chemotherapy according to the Stupp protocol in combination with a multi-target antiangiogenic drug and additional intrathecal chemotherapy using methotrexate. During treatment, the patient's tumor showed rapid progression. The chemotherapy with temozolomide was stopped and replaced with radiotherapy combined with tumor treating fields (TTF), the poly (adenosine diphosphate-ribose) polymerase (PARP) inhibitor niraparib, and anlotinib. After the radiotherapy was completed, the symptoms of increased intracranial pressure and epilepsy were well controlled. Considering the patient's tolerance to the treatment, the combined therapy of TTF and anlotinib was continued, and osimertinib, a third-generation epidermal growth factor receptor-tyrosine kinase inhibitor with good permeability of the blood-brain barrier, was added. The patient was regularly followed up and had no obvious adverse drug reactions. Head magnetic resonance imaging (plain scan + enhanced scan) suggested that the lesions were stable. For rapidly progressing glioblastomas or histological grade II/III IDHwt astrocytomas, the combination of TTF and a PARP inhibitor during radiotherapy may have a synergistic effect on tumor control and is well tolerated by patients.

Luminescent wearable biosensors based on gold nanocluster networks for "turn-on" detection of Uric acid, glucose and alcohol in sweat.

From the difficulty of awareness of abnormal concentrations of biochemical indexes in people's daily life come wearable sensing technologies. Recently, luminescent wearable biosensors are emerging with simple fabrication, easy use, cost-effectivity and reliability. But several challenges should be taken up, such as availability of varied analytes, high sensitivity, stability of enzymes, photostability, low signal noises and recyclability of sensors. Here, the Luminescent Wearable Sweat Tape (LWST) biosensor is developed via embedding multi-component nanoprobes onto microwell-patterned paper substrates of hollowed-out double-side tapes. The nanoprobes consist of responsive luminophores, enzyme-loaded gold nanocluster (AuNCs) nano-networks, which are wrapped by the switch, MnO2 nanosheets. The responsive luminophores are constructed by 3 substitutable components: enzymes (uricase, GOx and alcohol dehydrogenase) for molecular target recognition, glutathione-protected AuNCs (yellow, red and green) for luminescent signal output and polycations PAH for integration. MnO2 NSs as the switch can quench the emission of the AuNCs but degraded by the reductive product of incorporated enzymes. Thus, targeting analysts (uric acid, glucose and alcohol) can be dose-dependently detected through "turn-on" luminescence approach. After incorporating the nanoprobes into hollow-out tapes, the formed LWST biosensors can detect uric acid, glucose and alcohol in sweat with the help of a smartphone. Subsequently, we primarily apply them into human daily life scenario, sampling from dine parties, and the positive relationships of analyte intakes and the increase of analytes in sweat are significant with individual difference.

Contribution of Connexin Hemichannels to the Pathogenesis of Acute Lung Injury.

Connexin (Cx) family members form hemichannels (HCs) and gap junctions (GJs). Biological functions of Cx HCs have not been adequately characterized due to the inability to selectively target HCs or GJs. Recently, we developed a 6-mer peptide mimetic (P5) of the first extracellular loop of Cx43 and showed that it can block the permeability of HCs but not GJs formed by Cx43. In this study, we further characterized the HC blocking property of P5 and investigated the role of Cx HCs in acute lung injury (ALI). We found that P5 administration decreased HC permeability, in pulmonary microvascular endothelial cells, HepG2 cells, and even Cx43-deficient astrocytes, which express different sets of Cxs, suggesting that P5 is a broad spectrum Cx HC blocker. In addition, P5 reduced HC permeability of alveolar cells in vivo. Moreover, P5 decreased endotoxin-induced release, by vascular endothelial cells in vitro, of high mobility group box protein 1 (HMGB1), a critical mediator of acute lung injury (ALI), and reduced HMGB1 accumulation in bronchoalveolar lavage fluid (BALF) of mice subjected to intratracheal endotoxin instillation. Furthermore, P5 administration resulted in a significant decrease in the concentrations of ALT, AST, and LDH in the BALF, the accumulation of leukocytes in alveoli, and the mortality rate of mice subjected to ALI. Wright-Giemsa staining showed that P5 caused similar reductions of both neutrophils and monocytes in BALF of ALI mice. Together, these results suggest that Cx HCs mediate HMGB1 release, augment leukocyte recruitment, and contribute to ALI pathology.

The interplay between chromophore and protein determines the extended excited state dynamics in a single-domain phytochrome.

Phytochromes are a diverse family of bilin-binding photoreceptors that regulate a wide range of physiological processes. Their photochemical properties make them attractive for applications in optogenetics and superresolution microscopy. Phytochromes undergo reversible photoconversion triggered by the Z ⇄ E photoisomerization about the double bond in the bilin chromophore. However, it is not fully understood at the molecular level how the protein framework facilitates the complex photoisomerization dynamics. We have studied a single-domain bilin-binding photoreceptor All2699g1 (Nostoc sp. PCC 7120) that exhibits photoconversion between the red light-absorbing (Pr) and far red-absorbing (Pfr) states just like canonical phytochromes. We present the crystal structure and examine the photoisomerization mechanism of the Pr form as well as the formation of the primary photoproduct Lumi-R using time-resolved spectroscopy and hybrid quantum mechanics/molecular mechanics simulations. We show that the unusually long excited state lifetime (broad lifetime distribution centered at ∼300 picoseconds) is due to the interactions between the isomerizing pyrrole ring D and an adjacent conserved Tyr142. The decay kinetics shows a strongly distributed character which is imposed by the nonexponential protein dynamics. Our findings offer a mechanistic insight into how the quantum efficiency of the bilin photoisomerization is tuned by the protein environment, thereby providing a structural framework for engineering bilin-based optical agents for imaging and optogenetics applications.

Apatinib treatment for symptomatic pseudoprogression after standard treatment for glioblastoma multiforme: a case report.

Apatinib, a novel tyrosine kinase inhibitor, has anti-angiogenetic effect just as bevacizumab. Although bevacizumab has been used successfully in treating cerebral radiation necrosis, there has yet not any report on that apatinib can treat pseudoprogression with symptoms. Here we report a case of glioblastoma multiforme (GBM) patient with pseudoprogression after receiving the concurrent chemoradiotherapy, which was successfully treated by apatinib. A 51-year-old woman had multiple intracranial lesions (left parietal and right frontal), the primary left parietal lesion was surgically removed and was pathologically confirmed as glioblastoma (WHO grade IV). Then the patient received postoperative temozolomide with concurrent chemoradiotherapy. Three weeks after the radiotherapy, the patient experienced increased intracranial pressure and seizure. Magnetic resonance imaging (MRI) T1 enhancement examination showed an increase of abnormal enhancement range in the area of irradiation. After multiple disciplinary team (MDT) discussion, the patient was diagnosed with pseudoprogression after radiotherapy. Then she was given apatinib for 8 weeks at a dose of 500 mg qd. During the treatment period, the clinical symptoms and corresponding nerve images of the patient have been rapidly improved. In 12 months after the radiotherapy, progression of tumor in the primary site has not been discovered. Apatinib showed a good therapeutic effect and tolerance for the development of pseudoprogression advances with obvious symptoms.

Chromophorylation of cyanobacteriochrome Slr1393 from Synechocystis sp. PCC 6803 is regulated by protein Slr2111 through allosteric interaction.

Cyanobacteriochromes (CBCRs) are photochromic proteins in cyanobacteria that act as photosensors. CBCRs bind bilins as chromophores and sense nearly the entire visible spectrum of light, but the regulation of the chromophorylation of CBCRs is unknown. Slr1393 from Synechocystis sp. PCC 6803 is a CBCR containing three consecutive GAF (cGMP phosphodiesterase, adenylyl cyclase, and FhlA protein) domains, of which only the third one (Slr1393g3) can be phycocyanobilin-chromophorylated. The protein Slr2111 from Synechocystis sp. PCC 6803 includes a cystathionine ß-synthase (CBS) domain pair of an as yet unknown function at its N terminus. CBS domains are often characterized as sensors of cellular energy status by binding nucleotides. In this work, we demonstrate that Slr2111 strongly interacts with Slr1393 in vivo and in vitro, which generates a complex in a 1:1 molar ratio. This tight interaction inhibits the chromophorylation of Slr1393g3, even if the chromophore is present. Instead, the complex stability and thereby the chromophorylation of Slr1393 are regulated by the binding of nucleotides (ATP, ADP, AMP) to the CBS domains of Slr2111 with varying affinities. It is demonstrated that residues Asp-53 and Arg-97 of Slr2111 are involved in nucleotide binding. While ATP binds to Slr2111, the association between the two proteins gets weaker and chromophorylation of Slr1393 are enabled. In contrast, AMP binding to Slr2111 leads to a stronger association, thereby inhibiting the chromophorylation. It is concluded that Slr2111 acts as a sensor of the cellular energy status that regulates the chromophorylation of Slr1393 and thereby its function as a light-driven histidine kinase.

Alkaline pH shock enhanced production of validamycin A in fermentation of Streptomyces hygroscopicus.

Validamycin A (Val-A) is produced by Streptomyces as a secondary metabolite with wide agricultural applications of controlling rice sheath blight, false smut and damping-off diseases. The effect of alkaline pH shock on enhancing Val-A production and its mechanism were investigated. A higher yield of Val-A was achieved by NaOH shock once or several times together with faster protein synthesis and sugar consumption and alkaline pH shock can increase Val-A production by 27.43%. Transcription of genes related to amino acid metabolism, carbon metabolism and electron respiratory chain was significantly up-regulated, accompanied by the substantial increase of respiratory activity and glutamate concentration. Val-A production was promoted by a series of complex mechanisms and made a response to pH stress signal, which led to the enhancement of glutamate metabolism and respiration activity. The obtained information will facilitate future studies for antibiotic yield improvement and the deep revealment of molecular mechanism.

A FKBP5 mutation is associated with Paget's disease of bone and enhances osteoclastogenesis.

Paget's disease of bone (PDB) is a common metabolic bone disease that is characterized by aberrant focal bone remodeling, which is caused by excessive osteoclastic bone resorption followed by disorganized osteoblastic bone formation. Genetic factors are a critical determinant of PDB pathogenesis, and several susceptibility genes and loci have been reported, including SQSTM1, TNFSF11A, TNFRSF11B, VCP, OPTN, CSF1 and DCSTAMP. Herein, we report a case of Chinese familial PDB without mutations in known genes and identify a novel c.163G>C (p.Val55Leu) mutation in FKBP5 (encodes FK506-binding protein 51, FKBP51) associated with PDB using whole-exome sequencing. Mutant FKBP51 enhanced the Akt phosphorylation and kinase activity in cells. A study of osteoclast function using FKBP51V55L KI transgenic mice proved that osteoclast precursors from FKBP51V55L mice were hyperresponsive to RANKL, and osteoclasts derived from FKBP51V55L mice displayed more intensive bone resorbing activity than did FKBP51WT controls. The osteoclast-specific molecules tartrate-resistant acid phosphatase, osteoclast-associated receptor and transcription factor NFATC1 were increased in bone marrow-derived monocyte/macrophage cells (BMMs) from FKBP51V55L mice during osteoclast differentiation. However, c-fos expression showed no significant difference in the wild-type and mutant groups. Akt phosphorylation in FKBP51V55L BMMs was elevated in response to RANKL. In contrast, IκB degradation, ERK phosphorylation and LC3II expression showed no difference in wild-type and mutant BMMs. Micro-CT analysis revealed an intensive trabecular bone resorption pattern in FKBP51V55L mice, and suspicious osteolytic bone lesions were noted in three-dimensional reconstruction of distal femurs from mutant mice. These results demonstrate that the mutant FKBP51V55L promotes osteoclastogenesis and function, which could subsequently participate in PDB development.

[Clinical features and prognostic factors of brain metastasis from colorectal cancer].

OBJECTIVE: The aim of this study was to analyze the clinical features and prognostic factors in patients with brain metastasis from colorectal cancer (CRC). METHODS: Clinical materials of 45 colorectal cancer patients who developed brain metastasis were collected, and the data and follow-up data of those patients were retrospectively analyzed. RESULTS: Most brain metastases were from rectal cancer (64.4%), and 80.0% of the 45 cases had extracranial metastases. The most common extracranial metastatic site was the lung (57.8%), followed by the liver (35.6%). All the brain metastases in patients with liver metastases were supratentorial, while in contrast, 44.8% of the patients without liver metastasis had subtentorial metastasis, showing a significant difference between them (P<0.05). The interval time from diagnosis of CRC to the development of brain metastases in case of Dukes D stage was 12.0 months, significantly shorter than that in the cases of Dukes A stage (24.0 months), B (36.0 months) and C (29.0 months) (P<0.05). The interval time was also shorter in the patients who developed extracranial metastasis within one year than those more than one year (12.0 months vs. 38.0 months)( P<0.05). The median survival time of patients with brain metastasis from colorectal was 6.0 months, with a 1-year survival rate of 21.1% and 2-year survival rate of 3.3% only. Univariate analysis showed that the median survival of patients with a KPS score of ≥70 was 8.0 months, significantly higher than 2.0 months in those with a KPS score of <70 (P<0.05). The median survival of patients with one or two brain metastases was 8.0 months, significantly higher than 4.0 months of those with >2 brain metastases (P<0.05). The median survival time after diagnosis of brain metastasis was 4.0 months for those who received monotherapy (only steroids, only chemotherapy or only radiotherapy), significantly shorter than 10.0 months of patients who received chemoradiotherapy, and 12.0 months of those who underwent surgery (P<0.05). Comparing each two differently treated groups, the survival time of surgery combined with chemotherapy or radiotherapy group was significantly different from that of all of other groups (P<0.05). The median survival time of chemoradiotherapy group was longer than that of monotherapy, but the difference was not significant (P>0.05). Multivariate analysis showed that brain metastases >2 and treatment modality type are independent prognostic factors for survival. CONCLUSIONS: Patients initially diagnosed with a Dukes D stage primary colorectal tumor and occurrence of extracranial metastasis (especially, pulmonary metastasis) within one year are associated to an increased risk of brain metastases and have a shorter survival time. Most brain metastases in patients with liver metastases are supratentorial, while many patients without liver metastasis have subtentorial metastasis. Brain metastases >2 and the type of treatment modality are independent prognostic factors for survival. The prognosis of patients who received chemoradiotherapy is better than those treated only with chemotherapy or radiotherapy. Some subsets of patients may benefit from surgery plus chemotherapy/radiotherapy.

A novel periplasmic protein (Slr0280) tunes photomixotrophic growth of the cyanobacterium, Synechocystis sp. PCC 6803.

Cyanobacteria are among the main contributors to global photosynthesis and show a high degree of metabolic plasticity. Synechocystis sp. PCC 6803 can grow under photoautotrophic, photomixotrophic or photoheterotrophic conditions. We have characterized a novel periplasmic protein (Slr0280) that tunes the photomixotrophic growth of Synechocystis sp. PCC 6803. Slr0280 is a multi-domain protein consisting mainly of ß-sheets. Several proteins that interact with Slr0280 were identified via bacterial two-hybrid screening. Slr0280 may interact through its DUF2233 domain with partners that participate in sugar metabolism, thereby coordinating the respective regulations. When slr0280 was deleted, the mutant grew more slowly than wild-type in the presence of glucose, which is ascribed to the down-regulation of glycolysis, glycogen catabolism, oxidative pentose phosphate pathway, Calvin cycle and glucose utilization. A positive regulation of Slr0280 on these sugar catabolic enzymes was confirmed by transcript (qPCR) analyses. Based on these findings, we proposed a speculative model that Slr0280 plays a coordinating regulatory role in sugar metabolism.

Orange fluorescent proteins constructed from cyanobacteriochromes chromophorylated with phycoerythrobilin.

Cyanobacteriochromes are a structurally and spectrally highly diverse class of phytochrome-related photosensory biliproteins. They contain one or more GAF domains that bind phycocyanobilin (PCB) autocatalytically; some of these proteins are also capable of further modifying PCB to phycoviolobilin or rubins. We tested the chromophorylation with the non-photochromic phycoerythrobilin (PEB) of 16 cyanobacteriochrome GAFs from Nostoc sp. PCC 7120, of Slr1393 from Synechocystis sp. PCC 6803, and of Tlr0911 from Thermosynechococcus elongatus BP-1. Nine GAFs could be autocatalytically chromophorylated in vivo/in E. coli with PEB, resulting in highly fluorescent biliproteins with brightness comparable to that of fluorescent proteins like GFP. In several GAFs, PEB was concomitantly converted to phycourobilin (PUB) during binding. This not only shifted the spectra, but also increased the Stokes shift. The chromophorylated GAFs could be oligomerized further by attaching a GCN4 leucine zipper domain, thereby enhancing the absorbance and fluorescence of the complexes. The presence of both PEB and PUB makes these oligomeric GAF-"bundles" interesting models for energy transfer akin to the antenna complexes found in cyanobacterial phycobilisomes. The thermal and photochemical stability and their strong brightness make these constructs promising orange fluorescent biomarkers.