Nutrient enrichment and probiotics for sea urchin Anthocidaris crassipina larvae in captivity to promote large-scale aquaculture.

Sea urchin contains physiologically active substances, such as amino acids and unsaturated fatty acids, and an important aquatic organism. Purple sea urchin, one of the common edible sea urchins, is an important aquatic product. In order to supply the vast seafood market, large-scale aquaculture of sea urchins is very important. The aim of this study was to optimize the rearing of the Anthocidaris crassipina larvae enhancing the nutrition by mixing feed to improve their growth and survival. The survival rate of Chaetoceros muelleri feeding alone is only 40%. If the survival rate is improved through nutrient enrichment, the large-scale aquaculture of larvae can be promoted. The experiment was divided into two parts. Experiment 1: Two types of commonly used microalgae, Isochrysis galbana tml (I), C. muelleri (C) and two types of probiotics, Rhodopseudomonas palustris (R), and Saccharomyces cerevisiae (S) were used in the. Feeding amounts are 5000, 10,000, and 20,000 cell mL-1 , and the control group (N) did not eat. Experiment 2: C. muelleri 20,000 cell mL-1 was mixed with I. galbana tml, R. palustris (R) and S. cerevisiae (S) at 5000 and 10,000 cell mL-1 . After the experiment, body length, body width, stomach length, rudiment length, rudiment length, body composition, digestive enzymes and survival rate were measured to evaluate the best feed. The results showed that the mixed feeding of C. muelleri 20,000 cell mL-1 and R. palustris 5000 cell mL-1 can achieve the best development and survival of larval embryos and can promote metamorphosis into juveniles in the shortest time. The research results will be applied to the large-scale aquaculture of A. crassipina larvae to promote the diversity of aquaculture.

Co-expression of CD24 and Hsp70 as a prognostic biomarker for lung cancer.

Lung cancer is one of the most common malignant neoplasms worldwide. CD24 is a marker of tumor stem cells that plays an important role in tumorigenesis. Hsp70 is an important molecular chaperone. However, the co-expression and interaction of CD24 and Hsp70, as well as the significance for the prognosis of lung cancer are still unclear. The expression levels of CD24 and Hsp70 were detected by immunohistochemistry and their correlation was analyzed. The expression levels of CD24 mRNA and protein were examined using qRT-PCR and western blotting in SPCA1, A549, H1975, and H1650 cell lines. A CD24-overexpressing cell model was established. The interaction between CD24 and Hsp70 was verified by co-immunoprecipitation and western blotting. CD24 and Hsp70 expression were significantly higher in lung cancer tissues than in adjacent tissues (CD24: p=0.008; Hsp70: p<0.001). CD24 protein expression showed a positive correlation with lymph node metastasis, TNM stage, and vascular cancer thrombus. Hsp70 protein expression showed a positive correlation with differentiation, lymph node metastasis, and TNM stage. CD24 and Hsp70 high expression were also correlated with poor survival. The positive co-expression rate of CD24 and Hsp70 in lung cancer tissues was 52.7% (49/93). CD24 and Hsp70 expression in lung cancer were positively correlated (r=0.368, p<0.001), and co-immunoprecipitation was verified that both endogenous and exogenous CD24 co-precipitated with Hsp70 directly or indirectly. When Hsp70 inhibitor VER15508 was added to A549 cells, Hsp70 and CD24 protein expression were significantly decreased. The present study demonstrated that CD24 and Hsp70 were highly expressed in lung cancer tissues, and associated with invasion, metastasis, and poor survival. Hsp70 may regulate CD24 expression. Co-expression of CD24 and Hsp70 may be a prognostic biomarker for lung cancer.

Enantioselective Reductive Cyanation and Phosphonylation of Secondary Amides by Iridium and Chiral Thiourea Sequential Catalysis.

The combination of transition-metal catalysis and organocatalysis increasingly offers chemists opportunities to realize diverse unprecedented chemical transformations. By combining iridium with chiral thiourea catalysis, direct enantioselective reductive cyanation and phosphonylation of secondary amides have been accomplished for the first time for the synthesis of enantioenriched chiral α-aminonitriles and α-aminophosphonates. The protocol is highly efficient and enantioselective, providing a novel route to the synthesis of optically active α-functionalized amines from the simple, readily available feedstocks. In addition, the reactions are scalable and the thiourea catalyst can be recycled and reused.

A light-gated molecular brake with antilock and fluorescence turn-on alarm functions: application of singlet-state adiabatic cis → trans photoisomerization.

A light-gated molecular brake that displays both high braking power (∼10(7)) and high switching power (∼74%) is reported. The lower rate for brake-on than for brake-off switching of the pentiptycene rotor mimics the function of an antilock braking system (ABS) for vehicles on a loose surface. The brake is also armed with a fluorescence turn-on alarm for accidental deactivation of the ABS function by acids. All of these features are associated with the highly efficient singlet-state adiabatic cis → trans photoisomerization of the phenylstilbene chromophore.

The Effect of Feeding on Briareum violacea Growth, Survival and Larval Development under Temperature and Salinity Stress.

In recent years, climate change has often caused fluctuations in seawater salinity and temperature, which threaten the survival and growth of corals. Effectively improving the stress response to temperature and salinity changes in corals to prevent bleaching is one of the important issues. This study initially explored the use of artificial polyunsaturated fatty acids to assess the ability of Briareum violacea to slow bleaching, enhance growth, stabilize larval development and reduce antistress factors (superoxide dismutase and catalase) when they were exposed to temperature and salinity stress. The salinities used in the experiment were 25, 30, 35 and 40 psu, and the temperatures were 20, 25 and 30 °C. It was divided into two parts: Experiment 1-Effects of temperature and salinity and feeding on digestive enzymes, reproduction and stress response of B. violacea; Experiment 2-Effects of temperature and salinity and feeding on the settlement and survival of larvae. The results showed that the feeding treatment group reduced the superoxide dismutase, catalase and mortality of corals under stress and significantly improved larval development and larval settlement.

Effects of LED Light Illumination on the Growth, Digestive Enzymes, and Photoacclimation of Goniopora columna in Captivity.

Goniopora columna is a stony coral valued for its reef-building potential and its unique appearance. Thus, identifying the optimal culture conditions for G. columna would enable efficient cultivation and prevent the illegal exploitation of marine resources. Light sources are crucial for the growth of corals because zooxanthellae provide them with basic nutrients through photosynthesis. Different corals and zooxanthellae have different photoacclimation characteristics; therefore, selecting a suitable light wavelength remains the key inhibitor of coral maintenance in marine aquariums. Accordingly, this study investigated the effects of different light wavelengths on G. columna. It was illuminated for 6 or 12 h a day under white light, yellow light, red light (LR), green light (LG), blue light (LB), or purple light (LP) for 8 weeks. During the experiment, R(R; i.e., a formula feed that combines sodium alginate, protein and probiotics) of 5% (w/v) of G. columna tissue and skeletal dry weight was fed every day. Coral polyps were counted, zooxanthellae density, chlorophyll a concentration, specific growth rates, and survival rates were calculated; polyp stretching and contractile behaviors were observed; and body composition and digestive enzyme activity were analyzed. LB or LP (but not LG or LR) illumination for at least 6 h per day significantly promoted the growth, survival, protein content, and protease activity of the G. columna specimens. Furthermore, coral polyp extension reached 100% after 30 min of LP and LB light irradiation. Although no significant differences in the zooxanthellae density or chlorophyll a concentration were noted under various light wavelengths, significant reductions were detected in the absence of light. To achieve energy-efficient coral aquaculture with regard to G. columna cultivation, 6 h of LB or LP illumination per day can improve the growth.

Evaluation of Clove Extract for Drug Therapy of Ciliate Infection in Coral (Goniopora columna).

In recent years, ciliate infections have caused serious casualties to corals in the ocean. Infected corals die within a short period of time, which not only poses a threat to wild coral reefs, but also has a major impact on large scale aquaculture of coral. Clove is a kind of Chinese medicine with antifungal, antibacterial, antiviral, insecticidal, and other functions. Clove is a natural product. If it can be used in the treatment of coral ciliates, it will reduce this threat to the environment. The clove extract was diluted with sterile seawater to 500 ppm, 1500 ppm, 2500 ppm, 5000 ppm, 7500 ppm, and 10,000 ppm to carry out virulence test on ciliates. The results show that the LC50 value is 1500 ppm, which can cause the death of ciliates in 10 min without causing significant changes in G. columna SOD, CAT, chlorophyll a, and zooxanthellae. In addition, observation of tissue slices revealed that no ciliates and vacuum were found in the G. columna tissue after 10 min of medicated bathing. In summary, 1500 ppm of clove extract can be used for the treatment of coral ciliates.

A pentiptycene-derived molecular brake: photochemical E→Z and electrochemical Z→E switching of an enone module.

The synthesis and brakelike performance of a new molecular system (1) consisting of a pentiptycene rotor and a 2-methyleneindanone brake are reported. The rotation kinetics of the rotor was probed by both variable-temperature (1)H and (13)C NMR spectroscopy and DFT calculations, and the switching between the brake-on and brake-off states was conducted by a combination of photochemical and electrochemical isomerization. Because of the greater steric hindrance between the rotor and the brake units in the Z form ((Z)-1) than in the E form ((E)-1), rotation of the rotor is slowed down 500-fold at room temperature (298 K) on going from (E)-1 to (Z)-1, corresponding to the brake-off and brake-on states, respectively. The (E)-1→(Z)-1 photoisomerization in acetonitrile is efficient and reaches an (E)-1/(Z)-1 ratio of 11:89 in the photostationary state upon excitation at 290 nm, attributable to a much larger isomerization quantum efficiency for (E)-1 versus (Z)-1. An efficient (Z)-1→(E)-1 isomerization (96%) was also achieved by electrochemical treatment through the radical anionic intermediates. Consequently, the reversibility of the E-Z switching of 1 is as high as 85%. The repeated E-Z switching of 1 with alternating photochemical and electrochemical treatments is also demonstrated.

Toward a four-toothed molecular bevel gear with C2-symmetrical rotors.

The design, synthesis, conformational analysis, and variable-temperature NMR studies of pentiptycene-based molecular gears Pp(2)X, where Pp is the unlabeled (in 1H) or methoxy groups-labeled (in 1OM) pentiptycene rotor and X is the phenylene stator containing ortho-bridged ethynylene axles, are reported. The approach of using shape-persistent rotors of four teeth but C(2) symmetry for constructing four-toothed molecular gears is unprecedented. In addition, the first example of enantioresolution of chiral pentiptycene scaffolds is demonstrated. Density functional theory (DFT) and AM1 calculations on these Pp(2)X systems suggest two possible correlated torsional motions, geared rocking and four-toothed geared rotations, which compete with the uncorrelated gear slippage. The DFT-derived torsional barriers in 1H for rocking, four-toothed rotation, and gear slippage are approximately 2.9, 5.5, and 4.7 kcal mol(-1), respectively. The low energy barriers for these torsional motions result from the low energy cost of bending the ethynylene axles. Comparison of the NMR spectra of 1OM in a mixture of stereoisomers (1OM-mix) and in an enantiopure form (1OM-op) confirms a fast gear slippage in these Pp(2)X systems. The effect of the methoxy labels on rotational potential energy surface and inter-rotor dynamics is also discussed.

Feeding of a Scleractinian Coral, Goniopora columna, on Microalgae, Yeast, and Artificial Feed in Captivity.

Nutritional requirements are critical in the process of coral aquaculture. In addition to energy from symbiotic algae, corals obtain sufficient nutrition through heterotrophic feeding. Microalgae and yeast are commonly used as nutritional supplements for many aquaculture organisms. In addition, if artificial feed can match or improve upon the nutritional supplementation provided by microalgae and yeast in the case of G. columna, then feeding this coral would be markedly easier. Hence, this article preliminarily discusses feeds suitable for G. columna. In this study, artificial PUFA rich in animal protein (R), Saccharomyces cerevisiae, Isochrysis galbana tml, and Nannochloropsis oculate were fed to G. columna at quantities of 5% and 10% of body weight. Growth, survival, body composition, and digestive enzymes were assessed. Regarding body composition, the coral's protein content is higher than that of carbohydrate or fat; thus, evaluating the heterotrophic nutrition of G. columna by using protein absorption is appropriate. The protease content is also high in digestive enzymes. Protein content, protease activity, and specific growth rate were significantly higher in the R group than in other groups. The number of polyps in the groups fed R at 5% and 10% of body weight increased by 40.00 ± 2.43 and 47.33 ± 0.89 number, respectively, significantly greater increases than those achieved in the other groups (p < 0.05). Changes in body composition and digestive enzymes over a 24-h period were compared to determine the optimal feeding time. Protein content and protease activity increased markedly between 6:00 and 12:00. The experimental results suggest that R can improve the activity of G. columna digestive enzymes and their protein and lipid content in body tissue, shorten the cultivation time, and enhance the profitability of coral aquaculture.

Effects of Ciliate Infection on the Activities of Two Antioxidant Enzymes (SOD and CAT) in Captive Coral (Goniopora columna) and Evaluation of Drug Therapy.

Ciliate infection is a serious parasitic disease of coral. Infected coral rots and dies in a short time. In addition to killing corals by infecting them in the oceans, ciliate infection also poses a threat to corals farmed on a large scale. In this study, two antioxidant enzymes (SOD and CAT) were used to judge the stress response in Goniopora columna after infection, and KCl and H2O2 were used to evaluate the therapeutic effect. The results showed that SOD and CAT increased during the early stage of infection but decreased with the extension of infection time. In terms of drug therapy, it was found that the treatment of ciliate infection with 1.5% of KCl had no significant effect on SOD and CAT of G. columna. The morphological changes of zooxanthellae, chlorophyll a, and coral were not significant. H2O2 leads to a stress response and polyp contraction. In conclusion, 1.5% of KCl can be used in the selection of drugs to treat ciliate infection.

Selective and predicable amine conjugation sites by kinetic characterization under excess reagents.

The site selectivity for lysine conjugation on a native protein is difficult to control and characterize. Here, we applied mass spectrometry to examine the conjugation kinetics of Trastuzumab-IgG (Her-IgG) and α-lactalbumin under excess linker concentration ([L]0) based on the modified Michaelis-Menten equation, in which the initial rate constant per amine (kNH2 = Vmax/NH2/KM) was determined by the maximum reaction rate (Vmax/NH2) under saturated accessible sites and initial amine-linker affinity (1/KM). Reductive amination (RA) displayed 3-4 times greater Vmax/NH2 and a different panel of conjugation sites than that observed for N-hydroxysuccinimide ester (NHS) chemistry using the same length of polyethylene glycol (PEG) linkers. Moreover, faster conversion power rendered RA site selectivity among accessible amine groups and a greater tunable range of linker/protein ratio for aldehyde-linkers compared to those of the same length of NHS-linkers. Single conjugation with high yield or poly-conjugations with site homogeneity was demonstrated by controlling [L]0 or gradual addition to minimize the [L]0/KM ratio. Formaldehyde, the shortest aldehyde-linker with the greatest 1/KM, exhibited the highest selectivity and was shown to be a suitable probe to predict conjugation profile of aldehyde-linkers. Four linkers on the few probe-predicted hot spots were elucidated by kinetically controlled RA with conserved drug efficacy when conjugated with the payload. This study provides insights into controlling factors for homogenous and predictable amine bioconjugation.

Pentiptycene-derived light-driven molecular brakes: substituent effects of the brake component.

Five pentiptycene-derived stilbene systems (1 R; R = H, OM, NO, Pr, and Bu) have been prepared and investigated as light-driven molecular brakes that have different-sized brake components (1 H<1 OM<1 NO<1 Pr<1 Bu). At room temperature (298 K), rotation of the pentiptycene rotor is fast (k(rot)=10(8)-10(9)  s(-1)) with little interaction with the brake component in the trans form ((E)-1 R), which corresponds to the brake-off state. When the brake is turned on by photoisomerization to the cis form ((Z)-1 R), the pentiptycene rotation can be arrested on the NMR spectroscopic timescale at temperatures that depend on the brake component. In the cases of (Z)-1 NO, (Z)-1 Pr, and (Z)-1 Bu, the rotation is nearly blocked (k(rot)=2-6 s(-1)) at 298 K. It is also demonstrated that the rotation is slower in [D(6)]DMSO than in CD(2)Cl(2). A linear relationship between the free energies of the rotational barrier and the steric parameter A values is present only for (Z)-1 H, (Z)-1 OM, and (Z)-1 NO, and it levels off on going from (Z)-1 NO to (Z)-1 Pr and (Z)-1 Bu. DFT calculations provide insights into the substituent effects in the rotational ground and transition states. The molar reversibility of the E-Z photoswitching is up to 46%, and both the E and Z isomers are stable under the irradiation conditions.

A pentiptycene-derived light-driven molecular brake.

A room-temperature light-driven molecular brake (1), consisting of a pentiptycene rotator, a 3,5-dinitrophenyl brake, and a photoisomerizable ethenyl spacer, is reported. The rotation rates of the rotator differ by about 9 orders of magnitude between the brake-on (cis-1) and brake-off (trans-1) states.

General and chemoselective bisphosphonylation of secondary and tertiary amides.

With Tf2O as the activation reagent, a mild and general method has been developed for the bisphosphonylation of both secondary and tertiary amides. The protocol is highly efficient and chemoselective, and it tolerates a number of sensitive functional groups such as cyano, ester, and aldehyde groups.

An antilock molecular braking system.

A light-driven molecular brake displaying an antilock function is constructed by introducing a nonradiative photoinduced electron transfer (PET) decay channel to compete with the trans (brake-off) → cis (brake-on) photoisomerization. A fast release of the brake can be achieved by deactivating the PET process through addition of protons. The cycle of irradiation-protonation-irradiation-deprotonation conducts the brake function and mimics the antilock braking system (ABS) of vehicles.

A redox-gated slow-fast-stop molecular rotor.

A pentiptycene-derived p-phenylenediamine mimics a molecular double-rotor system that displays redox-dependent rotation rates for the amino rotors about the pentiptycene-amine C-N bond. The rotation is accelerated in the radical cation state but stopped in the di(radical cation) state. Electronic interplay of the two rotors is also discussed.

Synthesis of new halogenated pentiptycene building blocks.

The synthesis of central-ring halogenated pentiptycene phenols 12-14 and dihalogenated pentiptycenes 15-17 is reported. Their utilities as building blocks for preparing new pentiptycene-derived pi-conjugated systems through the Sonogashira, Heck, and Suzuki reactions are also demonstrated.