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Cacalia delphiniifolia and Cacalia hastata are edible wild plants in Japan. We found that these plants have anti-melanogenic activity in B16F10 mouse melanoma cells. Three furanoeremophilanes, cacalol (from C. delphiniifolia), dehydrocacalohastin, and cacalohastin (from C. hastata), were identified as the main active components. The genus Cacalia may be a good source of beneficial materials with anti-melanogenic effects.

Background Disruption of the indigenous microbiota is likely related to frailty caused by undernutrition. However, the relationship between undernutrition and the oral microbiota, especially normal bacteria, is not obvious. The aim of this study was to elucidate the associations of nutritional and oral health conditions with prevalence of bacteria and fungi in the oral cavity of older individuals. Methods Forty-one institutionalized older individuals with an average age ± standard deviation of 84.6 ± 8.3 years were enrolled as participants. Body mass index (BMI) and oral health assessment tool (OHAT) scores were used to represent nutritional and oral health status. Amounts of total bacteria, streptococci, and fungi in oral specimens collected from the tongue dorsum were determined by quantitative polymerase chain reaction (PCR) assay results. This study followed the STROBE statement for reports of observational studies. Results There was a significant correlation between BMI and streptococcal amount (ρ = 0.526, p p = 0.003). In logistic regression models, streptococcal amount was a significant variable accounting for “not undernutrition” [odds ratio 5.68, 95% confidential interval (CI) 1.64–19.7 (p = 0.06)]. On the other hand, participants with a poor oral health condition (OHAT ≥ 5) harbored significantly higher levels of fungi (p = 0.028). Conclusion Oral streptococci were found to be associated with systemic nutritional condition and oral fungi with oral health condition. Thus, in order to understand the relationship of frailty with the oral microbiota in older individuals, it is necessary to examine oral indigenous bacteria as well as etiological microorganisms.

The sustainability of energy and food supplies has come to represent a major concern throughout the world today. Greenhouse cultivation, an intensive food-production system, contributes fresh vegetables and fruits to the world food supply. Greenhouse crop yields and quality can be improved by microclimate controls powered by fuels and grid electricity inputs. Therefore, producing abundant and quality crops with improved energy efficiency has been pursued as a challenge to be addressed by researchers and practitioners. Although application of photovoltaics (PV) to greenhouses can reduce fuel and grid electricity consumption, PV inherently conflicts with cultivation because both photosynthesis and PV depend on sunlight availability. Various contrivances have been explored to enhance the compatibility of cultivation and PV power generation. This review describes important aspects of greenhouse cultivation, electricity demand in greenhouses, state-of-the-art of greenhouse PV systems, and PV shading effects on plants. Finally, prospects for energy-sustainable greenhouse PV technologies are presented.

North Pacific krill (Euphausia pacifica) contain 8R-hydroxy-eicosapentaenoic acid (8R-HEPE), 8R-hydroxy-eicosatetraenoic acid (8R-HETE) and 10R-hydroxy-docosahexaenoic acid (10R-HDHA). These findings indicate that E. pacifica must possess an R type lipoxygenase, although no such enzyme has been identified in krill. We analyzed E. pacifica cDNA sequence using next generation sequencing and identified two lipoxygenase genes (PK-LOX1 and 2). PK-LOX1 and PK-LOX2 encode proteins of 691 and 686 amino acids, respectively. Recombinant PK-LOX1 was generated in Sf9 cells using a baculovirus expression system. PK-LOX1 metabolizes eicosapentaenoic acid (EPA) to 8R-HEPE, arachidonic acid (ARA) to 8R-HETE and docosahexaenoic acid (DHA) to 10R-HDHA. Moreover, PK-LOX1 had higher activity for EPA than ARA and DHA. In addition, PK-LOX1 also metabolizes 17S-HDHA to 10R,17S-dihydroxy-docosahexaenoic acid (10R,17S-DiHDHA). PK-LOX1 is a novel lipoxygenase that acts as an 8R-lipoxygenase for EPA and 10R-lipoxygenase for DHA and 17S-HDHA. Our findings show PK-LOX1 facilitates the enzymatic production of hydroxy fatty acids, which are of value to the healthcare sector.

The integration of the photovoltaic (PV) energy in the greenhouse farm has raised concerns on the agricultural sustainability of this specific agrosystem in terms of crop planning and management, due to the shading cast by the PV panels on the canopy. The PV greenhouse (PVG) can be classified on the basis of the PV cover ratio (PVR), that is the ratio of the projected area of PV panels to the ground and the total greenhouse area. In this paper, we estimated the yield of 14 greenhouse horticultural and floricultural crops inside four commercial PVG types spread in southern Europe, with PVR ranging from 25 to 100%. The aim of the work is to identify the PVG types suitable for the cultivation of the considered species, based on the best trade-off between PV shading and crop production. The daily light integral (DLI) was used to compare the light scenarios inside the PVGs to the crop light requirements, and estimate the potential yield. The structures with a PVR of 25% were compatible with the cultivation of all considered species, including the high light demanding ones (tomato, cucumber, sweet pepper), with an estimated negligible or limited yield reduction (below 25%). The medium light species (such as asparagus) with an optimal DLI lower than 17 mol m−2 d−1 and low light crops can be cultivated inside PVGs with a PVR up to 60%. Only low light demanding floricultural species with an optimal DLI lower than 10 mol m−2 d−1, such as poinsettia, kalanchoe and dracaena, were compatible inside PVGs with a PVR up to 100%. Innovative cropping systems should be considered to overcome the penalizing light scenarios of the PVGs with high PVR, also implementing LED supplementary lighting. This paper contributes to identify the sustainable PVG types for the chosen species and the alternative crop managements in terms of transplantation period and precision agriculture techniques, aimed at increasing the crop productivity and adaptability inside the PVG agrosystems.

253 Background: An international staging collaboration for cancer of the prostate (STAR-CAP) was recently established and used for predicting oncological outcomes for patients with prostate cancer. However, the validation of STAR-CAP has not yet been performed in the Japanese cohort. Our study aimed to assess the STAR-CAP prognostic model for Japanese patients with prostate cancer and compare it to the CAPRA score for the prediction of biochemical recurrence (BCR) after radical prostatectomy. Methods: We retrospectively analyzed data from a multi-institution database of 1,645 patients who underwent radical prostatectomy between 2010 and 2020. We excluded patients with inadequate follow-up duration ( < 1 year), missing or incomplete perioperative clinical parameters relevant for STAR-CAP, and patients who received neoadjuvant therapy. BCR was defined as two consecutive prostate-specific antigen values ≥ 0.2 ng/mL. The 3-year BCR-free survival was assessed by Kaplan-Meier analyses and log-rank test. The performance of both models to predict 3-year BCR-free survival was assessed using the area under the curve (AUC). Results: The 3-year BCR-free survival ranged from 47.3% (high risk) to 93.5% (low risk) in the STAR-CAP classification ( p< 0.001) and from 62.5% (high risk) to 94.6% (low risk) in the CAPRA classification ( p< 0.001). AUCs for the 3-year BCR prediction were 0.756 and 0.784 for STAR-CAP and CAPRA, respectively. Conclusions: In Japanese patients, both STAR-CAP and CAPRA prognostic systems provide sufficient stratification and their predictive ability for 3-year BCR-free survival is comparable, with a small advantage for CAPRA (2.8%).

Surface-coated/uncoated epitaxial LiCoO2 film electrodes with (104) orientations were fabricated on SrRuO3(100)/Nb:SrTiO3(100) using pulsed laser deposition. Films with thicknesses of ~18 nm and flat surfaces with roughnesses of less than 1 nm were model systems for clarifying the kinetics of Li-ion transfer at the electrode/electrolyte interface. The Li-ion transfer characteristics at the interface between the LiCoO2 electrode and the electrolyte (LiPF6, ethylene carbonate + diethyl carbonate) were investigated by electrochemical impedance spectrometry. The charge-transfer resistance (Rct) of uncoated LiCoO2 increased from the early cycles when charged/discharged at 3.0–4.2 V. When charged/discharged at 3.0–4.5 V, the Rct of the uncoated LiCoO2 rapidly increased from the first charge. In contrast, the Rct of Li2ZrO3-coated LiCoO2 remained almost constant during the early cycles when charged/discharged either at 3.0–4.2 or at 3.0–4.5 V. The interfacial resistances of the coated and uncoated LiCoO2 electrodes were almost equal (~100 Ω cm2). The activation energy for charge transfer was lower for the coated LiCoO2 electrode compared to that for the uncoated electrode. The current-rate capability was significantly improved by surface coating even at high-voltage charge/discharge at 3.0–4.5 V. The charge transfer process is the rate-determining step of the charge/discharge reaction.

Euphausia pacifica is a good candidate for a resource of marine n-3 PUFA. However, few reports exist of the lipid and fatty acid composition of E. pacifica. To examine the potential of E. pacifica as a resource of marine n-3 PUFA, we analyzed E. pacifica oil. We extracted lipids from E. pacifica harvested from the Pacific Ocean near Sanriku, Japan. Lipid classes of E. pacifica oil were analyzed by TLC-FID and the fatty acid composition of the oil was analyzed by GC/MS. Free fatty acids and hydroxy-fatty acids were analyzed by LC/QTOFMS. The lipid content of E. pacifica ranged from 1.30% to 3.57%. The ratios of triacylglycerols, phosphatidylcholine, phosphatidylethanolamine and free fatty acids in E. pacifica lipids were 5.3–23.0%, 32.6–53.4%, 8.5–25.4% and 2.5–7.0%, respectively. The content of n-3 PUFA in E. pacifica lipids was 38.6–46.5%. We also showed that E. pacifica contains unusual fatty acids and derivatives: C16-PUFAs (9,12-hexadecadienoic acid, 6,9,12-hexadecatrienoic acid and 6,9,12,15-hexadecatetraenoic acid) and hydroxy-PUFAs (8-HETE and 10-HDoHE). E. pacifica is a good resource of marine n-3 PUFA. Moreover, E. pacifica can provide C16-PUFA and hydroxy-PUFAs.

This study introduces a novel algorithm to estimate the cumulated global radiation inside photovoltaic (PV) greenhouses at the desired time interval. The direct and diffuse radiation were calculated on several observations points (OPs) inside the PV greenhouse. The PV panels were assimilated to polygons that can overlap the sun path seen from a specific OP. The algorithm was tested in a greenhouse with 50% PV cover ratio on the roof. The results were showed as the percentage ratio of the cumulated yearly global radiation with and without PV array on the roof (GGR), and used to draw maps of light distribution on different canopy heights (from 0.0 to 2.0 m). The maps displayed the variability of the light distribution and the most adversely affected zones inside the PV greenhouse. The yearly GGR increased with the canopy height on the zones under the plastic cover (GGR from 59% at 0.0 m to 73% at 2.0 m), and decreased under the PV cover (GGR from 57% at 0.0 m to 40% at 2.0 m). Most zones close to the side walls and the gable walls were the least affected by shading on all canopy heights. The different light distribution on the canopy heights showed that the incident solar energy on the crop changes consistently, according to the growth stage of the plants. The algorithm can be applied to several PV greenhouse types and may provide a decision support tool for the identification of the most suitable plant species, based on their light requirements.

The capability and reversibility of surface-coated LiCoO2 (LCO) during charge/discharge with a layered structure change between O3 and H1−3 phases was investigated. A compound oxide phase with a thickness of several nanometers and a LiMO2 structure (M: Co, Al, or Zr) formed on the surface of the coated LCO. The discharge capacity (Q D) and discharge energy density (E D) at a current-rate of 0.05 C in the initial charge/discharge voltage range of 2.5–4.6 V were 228 mAh g−1 and 936 Wh kg−1, respectively. The Q D at the 1st cycle at 1 C was 213 mAh g−1, and the retention at the 101st cycle was 88.8%. The Q D and E D at 6 C were 204 mAh g−1 and 778 Wh kg−1. Operando XRD and impedance analyses revealed that the decrease in capacity by cycling was mainly due to the increased polarization, which was caused by the increase in the Li-ion intercalation resistance at the electrode/electrolyte interface. The O3/H1−3 phase transition was reversible even after the cycle-test. Most of the capacity in the O3/H1−3 transition region could be reversibly charged/discharged up to 100 cycles. The transition from H1−3 to O3 during discharging was responsive up to a current-rate of 6 C.

The use of renewable energy in modern greenhouse management is important to achieve efficient and sustainable food supplies for a world with increasing population. This study assessed the performance of a blind-type shading regulator that can automatically rotate semi-transparent photovoltaic (PV) blades installed on the greenhouse roof in response to sunlight variation. The PV blind oriented parallel to the roof partially blocked intense sunlight penetration into the greenhouse, but it transmitted sunlight during cloudy time by turning the blind bearing to be perpendicular to the roof. A stable irradiation environment is therefore producible in the greenhouse under variable sky conditions. Annual operations demonstrated that the blinds’ own generated electrical energy can sustain PV blind operation and produce surplus electrical energy. The PV blind electricity generation and sunlight availability for crops below the PV blind roof were calculated based on a mathematical model developed using theoretical sunlight parameters and the experimentally obtained PV blind system parameters. Assuming cloudless skies and threshold irradiance for blind rotation set at 500 W m−2, 13.0 and 12.3 kWh m−2 yr−1 surplus electrical energy can be generated, respectively, by north–south and east–west oriented model greenhouses. Cloudy skies reduce surplus electrical energy production by 50%, but PV blinds can supply greenhouse electrical energy demands partially or completely, depending on the degree of greenhouse electrification. Below the PV blinds, 8–10 MJ m−2 day−1 of insolation is expected to irradiate crops under actual sky conditions. This insolation is sufficient to cultivate major horticultural crops. Regulating the threshold irradiance level for PV blind turning can control the sunlight apportionment ratio for cultivation and electricity generation, thereby enabling sustainable energy–food dual production in a greenhouse.

The spherical micro-cells are a semi-transparent photovoltaic (PV) technology which can contribute to improve the sustainability of greenhouse systems. Previous prototypes were tested in laboratory conditions, but the size was not suitable for the greenhouse roof application. In this work, a new prototype has been developed and tested on a real greenhouse roof. The semi-transparent PV module (STM) was composed by 4800 spherical silicon micro-cells (1.2 mm diameter) sandwiched between glass plates and integrated on a greenhouse roof with 26.5° slope. The STM was 910 mm long and 610 mm wide to match the size of the greenhouse framework. The percentage of the STM area covered with micro-cells was 2.3%, reaching 9.7% considering the metallic conductors. The cell density was 2 cells cm−2 and the measured perpendicular light transmissivity of the semi-transparent area was 73%. The characteristics of the prototype were compared with those of a conventional planar multi-crystalline silicon module (CPM). The module conversion efficiency was steadily around 0.2% over wide incident sunlight angle. The micro-cells never completely eclipse the incident sunlight when observed from more than 1 m distance from the roof, keeping the eclipsing level at 9.7%. The yield factor of the STM was slightly higher than the CPM because of the isotropic properties of the spherical cells, which are able to use both the sky-incident and the ground-reflected irradiation for energy production, irrespective of the module slope. The prototype STM is promising for greenhouse roof applications and its performance can be improved by increasing the conversion efficiency.

An increasing population and limited arable land area endanger sufficient and variegated food supplies worldwide. Greenhouse cultivation enables highly intensive plant production and thereby enables the production of abundant fresh vegetables and fruits. The salient benefits of greenhouse cultivation are supported by ingenious management of crop environments, assisted by fossil fuel and grid electricity supplies. To reduce dependence on traditional energy resources, various studies have investigated exploitation of renewable energies for greenhouse environment management. Among them, solar photovoltaic (PV) technologies are anticipated to feed electrical energy to greenhouse appliances for microclimate control. This study proposes a venetian-blind-type shading system consisting of semi-transparent PV modules as blind blades based on micro-spherical solar cell technology to achieve greenhouse shading and electricity production concurrently. In response to the solar irradiance level, the PV blind inclination was altered automatically using a direct current (DC) motor driven by electrical energy generated by the PV blind itself. The PV blind was operated continuously during a five-month test period without outage. Moreover, the PV blind generated surplus electrical energy of 2125 kJ for blind system operations during the test period. The annual surplus energy calculated under the present experimental condition was 7.8 kWh m−2 year−1, suggesting that application of the PV blind to a greenhouse roof enables sunlight level control and electrical appliance operations in the greenhouse with a diminished fuel and grid electricity supply, particularly in high-insolation regions.

5-hydroxyeicosatetraenoic acid (5-HETE) and 5-hydroxyeicosapentaenoic acid (5-HEPE) are major metabolites produced by 5-lipoxygenase (5-LOX) from arachidonic acid (AA) and eicosapentaenoic acid (EPA). Effects of hydroxides on endothelial cells are unclear, although 5-LOX is known to increase at arteriosclerotic lesions. To investigate the effects of hydroxides on human umbilical vein endothelial cells (HUVECs), the cells were treated with 50 μM each of AA, EPA, 5-HETE, and 5-HEPE. Treatment of HUVECs with 5-HETE and 5-HEPE, rather than with AA and EPA, increased the nuclear translocation of NF-E2 related factor 2 (Nrf2) and upregulated the expression of heme oxygenase-1 and cystine/glutamate transporter regulated by Nrf2. Reactive oxygen species (ROS) generation was markedly elevated in HUVECs after treatment with 5-HETE and 5-HEPE, and the pretreatment with α-tocopherol abrogated ROS levels similar to those in the vehicle control. However, ROS generation was independent of Nrf2 activation induced by 5-HETE and 5-HEPE. 5-HETE was converted to 5-oxo-eicosatetraenoic acid (5-oxo-ETE) in HUVECs, and 5-oxo-ETE increased Nrf2 activation. These results suggest that 5-HETE works as an Nrf2 activator through the metabolite 5-oxo-ETE in HUVECs. Similarly, 5-HEPE works in the same way, because 5-HEPE is metabolized to 5-oxo-eicosapentaenoic acid through the same pathway as that for 5-HETE.

High-functional plastics have been attracted much attention over the various fields of industries. For evaluating their functionalities, it is essential to investigate low molecular weight components on the surface of the plastic because these components deeply affect their functionalities. In this study, multivariate analysis and g-ogram, a G-SIMS-related spectrum analysis method, were applied to the ToF-SIMS data of polyethylene glycol mixed model sample consisting of the two different average molecular weights, 600 u and 2000 u (PEG600 and PEG2000). As a result, the pure component spectra suggesting PEG600, PEG2000 and polydimethylsiloxane contaminant were discriminated by using multivariate curve resolution (MCR) under the condition of reducing the chemical noise such as heavily fragmented ions and contaminants. Moreover, the major peaks of the pure component spectrum suggesting PEG2000 in MCR were the characteristic fragment ions obtained by the g-ogram result for PEG2000. Therefore, it is indicated that the combination of MCR and g-ogram methods is a promising tool for evaluating the functionalities of the high-functional plastics. Copyright © 2014 John Wiley & Sons, Ltd.

While a number of heterologous expression systems have been reported for extracellular laccases, there are few for the intracellular counterparts. The Lentinula edodes intracellular laccase Lcc4 is an industrially potential enzyme with its unique substrate specificity. The heterologous production of the intracellular laccase, however, had been difficult because of its expression-dependent toxicity. We previously demonstrated that recombinant yeast cells synthesized and, interestingly, secreted Lcc4 only when they were suspended to an inducing medium in a high cell-density (J. Biosci. Bioeng., 113, 154-159, 2012). The high cell-density system was versatile and applicable to other difficult-to-express secretory proteins. Nevertheless, the system's great dependence on aeration, which was a practical obstacle to scale-up production of the enzyme and some other proteins, left the secretion pathway and enzymatic properties of the Lcc4 uncharacterized. In this report, we demonstrate a successful production of Lcc4 by applying a jar-fermentor to the high cell-density system. The elevated yield (0.6 mg L(-1)) due to the sufficient aeration allowed us to prepare and purify the enzyme to homogeneity. The enzyme had been secreted as a hyper-glycosylated protein, resulting in smear band-formations in SDS-PAGE. The amino acid sequencing analysis suggested that the N-terminal 17 residues had been recognized as a secretion signal. The recombinant enzyme showed similar enzymatic properties to the naturally occurring Lcc4. The characteristics of the scale-upped expression system, which includes helpful information for the potential users, have also been described.

Peptide vaccines have two fundamental weak points, namely low antigenicity and MHC-restriction. In our previous study, we proposed the design of vaccine peptide to overcome these weakpoints. The vaccine was constructed in the following order, N-terminal, Arg-Gly-Asp (RGD), T-cell epitope peptide, di-lysine linker (KK) to B-cell epitope peptide. Although the vaccine peptide can basically induce B-cell epitope peptide specific antibodies to the host without immune adjuvants via intraperitoneal, subcutaneous and intranasal administration, some peptide antigens require adjuvants for antibody induction. In this study, we propose a novel protocol to enhance the immunogenicity of the peptide utilizing the host immune response to a conventional toxoid vaccine, which are lymphocyte activities to the T-cell epitope peptide. We selected multiagretope-type T-cell epitopes from diphtheria toxoid, a conventional vaccine antigen, and a part of amyloid-beta peptide (Aβ) as a B-cell epitope. The conventional toxoid vaccine was immunized before the peptide immunization. Using this protocol, we succeeded in the enhancement of the anti-Aβ antibodies induction by intranasal immunization without any immune adjuvants in C57BL/6 and Balb/c mice. Furthermore, the vaccine peptide induced the transformation of peripheral blood lymphocytes collected from healthy volunteers carrying immunities to diphtheria toxoid. These results suggested that our peptide vaccines with the novel protocol would provide an effective method for antibody induction.

Polyporus brumalis ibrc05015 is a strain capable of high laccase (Lac) production. Among several inducers, 0.25 mM copper was most effective for Lac production. One of the Lacs induced by copper was PbLac1, and its transcription was induced within 60 min after copper addition. The promoter region of pblac1 contained six putative metal response elements and one Ace1 consensus cis-element. We cloned the P. brumalis PbAce1 transcription factor, a homologue of Saccharomyces cerevisiae transcription factor Ace1, which regulates metallothionein genes in response to excess copper. PbAce1 complemented the function of Ace1 in an S. cerevisiae Δace strain. The conserved N-terminal copper-fist DNA binding domain of PbAce1 was required for complementation. In the PbAce1 complemented Δace1 strain, the pblac1 promoter was constitutively expressed at a high level, independent of copper concentration. PbAce1 has two Cys-rich repeat motifs (PbC1 and PbC2), which are similar to the Cys-rich repeat domain in metallothionein proteins, and are uniquely conserved in the C-terminal domain of basidiomycetous Ace1 sequences. These C-terminal domains could be involved in copper sensing and concentration-dependent Lac production in basidiomycetous fungi.

Light energy irradiated on plants cultivated in a plant factory with artificial lighting (PFAL) is supplied from electrical energy through lamps that convert electricity into light. Plants exploit light as the energy source for driving photosynthesis. Plants also use light as a signal to trigger photomorphogenesis. An efficient use of both light and electrical energy is necessary for PFALs. This chapter presents a discussion of efficiencies of conversions step-by-step, from electrical input energy to the final chemical energy stored primarily as carbohydrates in crops in a PFAL.

Light-emitting diodes (LEDs) have been exploited from laboratory plant researches to commercial cultivations in greenhouses or plant factories with artificial lighting (PFAL). Benefits of LEDs for plant cultivation are their ability to emit thermal-free monochromatic light, solidity, minute size, minimal electricity consumption, longevity, and controllability of light emission. Varieties of LED lamps are available in the market, specialized respectively for PFAL, greenhouse, and laboratory applications. Those commercial products are well designed and useful, but users also can design and fabricate crop-specific LED lighting systems according to their purposes. To maximize the merits of LEDs for plant cultivations, the knowledge about fundamental principles of LED engineering would help growers. In this chapter, the light emission from a semiconductor p–n junction, the regulation of emission, heat treatments, and the regulation of light distribution on a culture bed are described.

The use of renewable energy for greenhouse environment control to replace or reduce the consumption of fuel and power-line electricity is an important objective for sustainable greenhouse crop production. This study was undertaken to apply a solar photovoltaic (PV) array to supply electricity for greenhouse environment control. The PV array was mounted inside the south roof of an east–west oriented single-span greenhouse, in which Welsh onion (Allium fistulosum L.) was cultivated hydroponically. Effects of PV-array shading on the Welsh onion growth were assessed. Two PV-array formations were tested: straight-line and checkerboard. For each arrangement, the PV array covered 12.9% of the greenhouse roof area. Beside the PV greenhouse, a control greenhouse was built with identical dimensions and orientation to those of the PV greenhouse. Welsh onion was cultivated also in the control greenhouse. The straight-line arranged PV-array (PVs array) cast shadows on a specific area of the cultivated plants continuously during the growth period. The fresh weight (FW) and dry-matter weight (DW) of Welsh onion cultivated under the PVs array shadow were significantly less than those of Welsh onion cultivated in the control greenhouse. The checkerboard PV-array (PVc array) cast shadows in the PVc greenhouse intermittently during growth. Consequently, the inhibitory effects of the PV-array shading on the FW and DW accumulations of Welsh onion were diminished. The electrical energy generated by the PVc array was comparable to that of the PVs array, which is another advantage of the PVc array.

Yeast's extracellular expression provides a cost-efficient means of producing industrially useful recombinant proteins. However, depending on the protein to be expressed, the production results in a poor yield, which is occasionally accompanied with loss of the expression plasmid and hence hampered growth of the host in the inducing medium. Here we propose an alternative approach, high cell-density expression, to improve the yield of a certain range of so-called difficult-to-express proteins. In this expression system, recombinant yeast cells resting in stationary phase (OD(660)=3-4) are suspended in a small aliquot of inducing medium to form a high cell-density culture (e.g., OD(660)=15). When applied to the yeast strains harboring Lentinula edodes laccase (Lcc1 or Lcc4) expressing plasmids, the high cell-density system allowed the host cells to synthesize elevated amounts of the laccase which resulted in >1000- to 6000-fold higher yield than those synthesized in a classical growth-associated manner. The resting cells required aerobic agitation for the maximum production. The production system also worked for other foreign enzymes but not for beta-galactosidase from Aspergillus oryzae or Escherichia coli, likely suggesting an involvement of chaperons that act on a certain range of secretory proteins.