The effect of sugar solution type,sugar concentration and viscosity on the imbibition and energy intake rate of bumblebees

Nectar is an essential resource for bumblebees and many other flower-visiting insects. The main constituents of nectar are sugars, which vary in both composition and concentration between plant species. We assessed the influence of sugar concentration, sugar solution viscosity and sugar solution composition on the imbibition and energy intake rate of bumblebees, Bombus impatiens Cresson (Hymenoptera: Apidae). To do this, we measured their rate of solution intake for 49 different sugar solution treatments, which varied in both sugar composition and concentration. In general, the imbibition rates of bumblebees were found to increase with increasing sugar concentration, probably due to their preference for high sugar concentrations, up to a concentration of 27% (w/w), at which point solutions reached a threshold viscosity of approximately 1.5–1.6 mPa.s. Above this threshold, the increasing viscosity of the solutions physically inhibited the imbibition rates of bees, and imbibition rate began to decrease as the concentration increased. Nevertheless, bumblebee energy intake rate increased with increasing concentration up to about 42–56%. Although we found that sugar solution composition had an impact on both imbibition and energy intake rate, its effect was not as straightforward as that of sugar concentration and viscosity.     

Study of potential binding of biologically important sugars with a dinuclear cobalt(II) complex

The investigation of the sugar–metal ion interactions remains one of the main objectives of carbohydrate coordination chemistry because the interactions between metal ions and carbohydrates are involved in many biochemical processes. The potential binding interaction between a five-coordinate dinuclear cobalt(II) complex, Na₂[Co₂(tcdc)(μ-OAc)] (1) [Na₅tcdc = Sodium-N,N,N′,N′-tetrakis(sodium carboxylate methyl)-2,6-diaminocresolate] and biologically important sugar substrates (d-glucose, d-xylose, and d-mannose) has been studied. In alkaline media, the complex 1 shows an excellent chelating ability toward these sugar substrates. A combined approach of FTIR and UV–vis spectroscopic investigations shows that the complex forms a 1:1 complex/substrate-bound product. UV–vis spectra indicate a significant blue-shift of the absorption maximum of metal complex during carbohydrate coordination highlighting the sugar binding ability of complex 1. The apparent binding constants of the substrate-bound cobalt(II) complexes have been determined from the UV–vis titration experiments.     

BSA–boronic acid conjugate as lectin mimetics

We report bovine serum albumin (BSA)–boronic acid (BA) conjugates as lectin mimetics and their glyco-capturing capacity. The BSA–BA conjugates were synthesized by amidation of carboxylic acid groups in BSA with aminophenyl boronic acid in the presence of EDC, and were characterized by Alizarin Red S (ARS) assay and SDS–PAGE gel. The BSA–BA conjugates were immobilized onto maleimide-functionalized silica beads and their sugar capturing capacity and specificity were confirmed by ARS displacement assay. Further, surface plasmon resonance (SPR) analysis of the glyco-capturing activity of the BSA–BA conjugates was conducted by immobilizing BSA–BA onto SPR gold chip. Overall, we demonstrated a BSA–BA-based lectin mimetics for glyco-capturing applications. These lectin mimetics are expected to provide an important tool for glycomics and biosensor research and applications.     

Extraction,characterization and spontaneous emulsifying properties of pectin from sugar beet pulp

The effects of organic acid extractants on the yield and characteristics of pectin from sugar beet pulp were investigated with citric acid, malic acid and lactic acid at different pH (1.5 and 2.0) and time (1 h and 2 h). The results demonstrated that the yields of pectins were directly correlated with the decrease of pH and reaction time, and the optimum yield of 17.2% was obtained at pH 1.5 and 2 h. Furthermore, the acid type also affected the physicochemical characteristics of pectin, especially on the esterification degree (42–71), galacturonic acid content (60.2–77.8%), emulsion activity (35.2–40.1%) and emulsion stability (62.1–79.4%), and a relatively single pectin mainly consisted of homogalacturonan could be obtained under a suitable reaction condition, which was an excellent crude material for the production of emulsion activity.     

UV-B辐射对拟南芥种子萌发和幼苗生长的影响

以哥伦比亚生态型(Columbia-0)拟南芥(Arabidopsis thaliana)为实验材料, 人工模拟UV-B辐照处理拟南芥种子, 统计其发芽势和发芽率, 测定根长、株高、叶绿素、可溶性糖、可溶性蛋白以及丙二醛(MDA)含量, 研究UV-B辐照处理对拟南芥种子萌发和幼苗生长的影响。研究结果表明, 低剂量的UV-B辐照可以促进拟南芥种子萌发和幼苗生长, 并且最佳辐照剂量为1.0 kJ·m^–2(P<0.05), 此时的发芽势、发芽率、根长、株高、叶绿素、可溶性糖及可溶性蛋白的含量均达到最大, 而丙二醛的含量变化则不明显(P>0.05)。当辐照剂量大于1.0 kJ·m^–2时, 促进作用逐渐变小, 并且随着辐照剂量的增加, 表现出了抑制作用。实验结果表明, 适当剂量的UV-B辐照在一定程度上可以促进拟南芥种子萌发和幼苗生长, 而过高剂量的辐照则对其产生抑制或损伤作用。     

Label-free THz sensing of living body-related molecular binding using a metallic mesh

We have demonstrated label-free THz sensing of living body-related molecular binding using a thin metallic mesh and a polyvinylidene difluoride (PVDF) membrane. Metallic meshes in the THz region are designed for anomalous transmission phenomena derived from a resonant excitation of surface waves. Additionally, they are designed to have a sharp dip in transmittance. The metallic mesh is very sensitive to a change of the refractive index of materials attached to the metallic mesh. In this paper, we report sensing of interactions between lectin and sugar using this technique. We found that the dip frequency shift, transmittance attenuation of the dip frequency, and peak shift of the derivative spectrum of the phase shift depend on the bonding amount of lectin–sugar interactions. We also applied this technique to detect avidin–biotin interactions, leading to the detection of a small amount of biotin (0.17 pg/mm²).     

Structural characterization of a rhamnose-binding glycoprotein (lectin) from Spanish mackerel (Scomberomorous niphonius) eggs

A rhamnose-binding glycoprotein (lectin), named SML, was isolated from the eggs of Spanish mackerel (Scomberomorous niphonius) by affinity and ion-exchange chromatographies. SML was composed of a non-covalently linked homodimer. The SML subunit was composed of 201 amino acid residues with two tandemly repeated domains, and contained 8 half-Cys residues in each domain, which is highly homologous to the N-terminal lectin domain of calcium-independent α-latrotoxin receptor in mammalian brains. Each domain has the same disulfide bonding pattern; Cys10–Cys40, Cys20–Cys99, Cys54–Cys86 and Cys67–Cys73 were located in the N-terminal domain, and Cys108–Cys138, Cys117–Cys195, Cys152–Cys182 and Cys163–Cys169 were in the C-terminal domain. SML was N-glycosylated at Asn168 in the C-terminal domain. The structure of the sugar chain was determined to be NeuAc-Galβ1-4GlcNAcβ1-2Manα1-6-(NeuAc-Galβ1-4GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc-Asn.     

Cryo-responses of two types of large unilamellar vesicles in the presence of non-permeable or permeable cryoprotecting agents

Cryo-responses of two types of large unilamellar vesicles (LUV) that were made from either egg yolk l-α-phosphatidylcholine (EPC) or 1,2-dipalmitoyl-rac-glycero-3-phosphocholine (DPPC), in the presence of non-permeable or permeable cryoprotective agents (CPA) was investigated. Partial ternary phase diagrams of CPA–salt–water with specific CPA to salt ratio (R), were constructed to estimate the phase volume of ice and unfrozen matrix of the LUV dispersion, which could aid in understanding the mechanistic actions of CPA. Leakage of both EPC and DPPC LUV was reduced if the sugar concentrations are above 10% (w/w) for disaccharides and 5% (w/w) for monosaccharides. Above these sugar concentrations, non-permeable CPA were more effective in preventing leakage of DPPC LUV than in EPC LUV. Below these sugar concentrations, EPC and DPPC LUV with limited mobility in the remaining unfrozen matrix were more likely to approach and interact with one and another, which were not anticipated when the LUV were completely embedded in the ice matrix. In the presence of Me₂SO or EG, EPC LUV that had been subjected to freezing and thawing processes were protected from leakage. At room temperature, Me₂SO and EG were detrimental to the DPPC LUV. This study suggests that the choice of CPA for cell cryopreservation depends on the type of phospholipids in plasma membranes, which vary in their acyl chain length and gel–liquid crystal phase transition temperature.     

Synthesis of C-glycosylated amino acids by hetero-Diels-Alder addition of ethyl 2-nitrosoacrylate to exo-glycals

C-Glycoamino acids bearing a variety of sugar moieties were prepared by the hetero-Diels–Alder addition of ethyl 2-nitrosoacrylate to exo-glycals. The reaction proceeds smoothly to yield spirocyclic oxazines that can be converted into useful products by several hydrogenolytic techniques.     

Studies on the synthesis and the antimicrobial and antioxidant activities of a novel class of fluorescein-based glycosides

Facile glycosylation of a fluorescein diol derivative with per-O-acetyl/benzoyl sugar derivatives using BF₃·Et₂O catalyst resulted in the formation of the expected glycosides in 54–66% yield. The biological screening of the glycosides against different microbes shows good inhibitory activity. The antioxidant activity of the fluorescein-based glycosides shows remarkable inhibition (IC₅₀ ∼80%).     

Crystal structures of (2R,4R)-2-(polyhydroxyalkyl)-1,3-thiazolidine-4-carboxylic acids: condensation products of l-cysteine with d-hexoses

We report herein the first crystal structures of (4-carboxy-1,3-thiazolidin-2-yl)pentitols [2-(polyhydroxyalkyl)thiazolidine-4-carboxylic acids], condensation products of l-cysteine with d-galactose and d-mannose: 2-(d-galacto-pentahydroxypentyl)thiazolidine-4-carboxylic acid hydrate, Gal-Cys·H₂O (1), and 2-(d-manno-pentahydroxypentyl)thiazolidine-4-carboxylic acid hydrate, Man-Cys·H₂O (2). In 1 and 2 the compounds crystallize as zwitterions, with the carboxylic groups deprotonated and the thiazolidine N atoms protonated. The sugar moiety and carboxylate group are in a cis configuration relative to the thiazolidinium ring, which adopts different conformation: twisted (T) on C^β–S in 1, and S-puckered envelope (E) in 2. The carbon chain of the galactosyl/mannosyl moiety remains in an extended zig-zag conformation. The orientation of the sugar O2 atom with respect to the thiazolidinium S and N atoms is trans–gauche in 1 and gauche–gauche in 2. The molecular conformation is stabilized by the intramolecular N–H?O_Cys contacts in both 1 and 2 and by the additional N–H?O_Man interaction in 2. The crystal packing of orthorhombic 1 and monoclinic 2 is determined mainly by N/O/C–H?O hydrogen bonds forming ribbons linked to each other by direct and water-mediated O/C–H?O/S contacts.     

罗布麻对不同浓度盐胁迫的生理响应

利用网室盆栽实验, 研究不同浓度的NaCl(100–400 mmol·L^–1)胁迫对罗布麻(Apocynum venetum)生长及生理特性 的影响。结果表明, 100 mmol·L^–1NaCl处理显著降低了罗布麻植株的鲜重, 但对其干重影响不大; 随着盐浓度继续增加, 罗布麻鲜重和干重显著下降。在盐胁迫下, 罗布麻叶片内的丙二醛含量、电解质渗漏率、根部和地上部Na⁺的含量明显增加, K⁺的含量随着盐离子浓度的增加而降低。盐胁迫显著降低了地上部Ca²⁺的含量, 而对根部Ca²⁺的含量没有影响。植株K⁺/Na⁺和Ca²⁺/Na⁺比值随着盐胁迫强度的增加而降低。盐胁迫显著促进了罗布麻根部对K⁺和Ca²⁺的选择性吸收及对K⁺的选择性运输。当NaCl浓度小于或等于200 mmol·L^–1时, 随着盐离子浓度的增加, 罗布麻叶片内的脯氨酸和可溶性糖积累显著增加,而当NaCl浓度大于200 mmol·L^–1时, 这2种有机溶质含量显著下降。总体上, 罗布麻通过积累无机离子、合成有机溶质及维持较高的K⁺、Ca²⁺选择性吸收和运输来适应一定浓度(≤200 mmol·L^–1NaCl)的盐胁迫。     

New dinuclear copper(II) and zinc(II) complexes for the investigation of sugar–metal ion interactions

We have studied the binding interactions of biologically important carbohydrates (d-glucose, d-xylose and d-mannose) with the newly synthesized five-coordinate dinuclear copper(II) complex, [Cu₂(hpnbpda)(μ-OAc)] (1) and zinc(II) complex, [Zn₂(hpnbpda)(μ-OAc)] (2) [H₃hpnbpda = N,N′-bis(2-pyridylmethyl)-2-hydroxy-1,3-propanediamine-N,N′-diacetic acid] in aqueous alkaline solution. The complexes 1 and 2 are fully characterized both in solid and solution using different analytical techniques. A geometrical optimization was made of the ligand H₃hpnbpda and the complexes 1 and 2 by molecular mechanics (MM+) method in order to establish the stable conformations. All carbohydrates bind to the metal complexes in a 1:1 molar ratio. The binding events have been investigated by a combined approach of FTIR, UV–vis and ¹³C NMR spectroscopic techniques. UV–vis spectra indicate a significant blue shift of the absorption maximum of complex 1 during carbohydrate coordination highlighting the sugar binding ability of complex 1. The apparent binding constants of the substrate-bound copper(II) complexes have been determined from the UV–vis titration experiments. The binding ability and mode of binding of these sugar substrates with complex 2 are indicated by their characteristic coordination induced shift (CIS) values in ¹³C NMR spectra for carbon atoms C1, C2, and C3 of sugar substrates.     

氮调控对盐环境下甜菜功能叶光系统Ⅱ荧光特性的影响

采用盆栽试验方法,以NaCl为盐分模拟不同盐度环境,研究了施氮(N)对盐环境下生长的甜菜(Beta vulgaris)功能叶光系统Ⅱ (PSⅡ)荧光特性的影响及光合色素含量的变化.结果表明:在轻度、中度及重度盐环境下,施N均能增大PSⅡ最大光化学效率(Fv/Fm)、PSⅡ潜在活性(Fv/Fo)、PSⅡ实际光量子产量(Y(Ⅱ))、非调节性能量耗散的量子产量(Y(NO))、相对电子传递速率(ETR)及光化学猝灭系数(qp),且在适宜的施N范围内(0-1.2 g·kg-1)上述参数随施N量的增加而增大.各叶绿素荧光参数光响应的结果表明,随着光强的增加,各处理下调节性能量耗散的量子产量(KNPQ))、ETR及非光化学猝灭系数(NPQ)旱上升趋势,相反,Y(Ⅱ)、Y(NO)及qp则呈下降趋势,在有效的光强范围内(0-1 000 μmol·m-2·s-1)施N提高了甜菜功能叶PSⅡ反应中心的开放程度,并且在高光强下调节PSⅡ耗散掉过剩的光能以避免对其反应中心造成伤害.各盐度环境下施N也显著增加了甜菜功能叶叶绿素与类胡萝卜素含量,增大了叶绿素a/叶绿素b值,且叶绿素与类胡萝卜素含量随施N水平的增加而增加.说明盐环境下施N能够增强甜菜功能叶PSⅡ的活性,提高PSⅡ光能利用率,从而增强其对盐渍环境的适应性.     

Increased tissue kallikrein amidase activity in urine of patients with type 1 diabetes under insulin therapy, and in those with gestational diabetes mellitus not under insulin therapy

Human tissue kallikrein (hK1) is reduced in hypertension, cardiovascular and renal diseases. There is little information on the participation of hK1 in type 1 diabetes mellitus (DM), type 2 DM, and gestational diabetes mellitus (GDM), respectively. The aim of this study was to evaluate the roles of insulin and hyperglycemia on urinary hK1 activity in type 1 DM and in GDM. Forty-three type 1 DM patients (5–35 years, disease duration ?5 years, receiving insulin, HbA_1c > 7.6%) were selected. Forty-three healthy individuals, paired according to gender and age, were used as controls. Thirty GDM patients (18–42 years, between the 24th and 37th week of pregnancy, recently diagnosed, not under insulin therapy) were also selected. Thirty healthy pregnant (18–42 years, between the 24th and 37th week of pregnancy) and 30 healthy non-pregnant women (18–42 years) were selected as controls. Random midstream urine was used. hK1 amidase activity was estimated with D-Val-Leu-Arg-Nan substrate. Creatinine was determined by Jaffe’s method. hK1 specific amidase activity was expressed as μM/(min mg creatinine) to correct for differences in urine flow rate. hK1 specific amidase activity was significantly higher in the urine of type 1 DM than in controls, and in the urine of GDM patients than in healthy pregnant women and healthy non-pregnant women, respectively. The data suggest that hyperglycemia, rather than insulin, is involved in the mechanism of increased hK1 specific amidase activity in both type 1 DM and GDM patients, respectively.     

Obesity and Sugar‐sweetened Beverages in African‐American Preschool Children: A Longitudinal Study

A representative sample of 365 low‐income African‐American preschool children aged 3–5 years was studied to determine the association between sugar‐sweetened beverage consumption (soda, fruit drinks, and both combined) and overweight and obesity. Children were examined at a dental clinic in 2002–2003 and again after 2 years. Dietary information was collected using the Block Kids Food Frequency Questionnaire. A BMI score was computed from recorded height and weight. Overweight and obesity were defined by national reference age‐sex specific BMI: those with an age‐sex specific BMI ≥85th, but <95th percentile as overweight and those with BMI ≥95th age‐sex specific percentile as obese. The prevalence of overweight was 12.9% in baseline, and increased to 18.7% after 2 years. The prevalence of obesity increased from 10.3 to 20.4% during the same period. Baseline intake of soda and all sugar‐sweetened beverages were positively associated with baseline BMI z‐scores. After adjusting for covariates, additional intake of fruit drinks and all sugar‐sweetened beverages at baseline showed significantly higher odds of incidence of overweight over 2 years. Among a longitudinal cohort of African‐American preschool children, high consumption of sugar‐sweetened beverages was significantly associated with an increased risk for obesity.     

Identification of proteoglycan from salmon nasal cartilage

There has been no structural information about the core protein of salmon nasal cartilage proteoglycan although its physiological activities have been investigated. Internal amino acid sequencing using nano-LC/MS/MS revealed that the salmon proteoglycan was aggrecan. Primer walk sequencing based on the amino acid information determined that the salmon aggrecan cDNA is comprised of 4207 bp nucleotides predicted to encode 1324 amino acids with a molecular mass of 143,276. It exhibited significant similarities to predicted pufferfish aggrecan, zebrafish similar to aggrecan, zebrafish aggrecan, bovine aggrecan and human aggrecan isoform 2 precursor; whose amino acid identities were 56%, 55%, 49%, 31% and 30%, respectively. Salmon cartilage aggrecan had globular domains G1, G2 and G3 as in mammalian aggrecans. Neither the putative keratan sulfate attachment domain enriched with serine, glutamic acid and proline, nor the putative chondroitin sulfate attachment domain with repeating amino acid sequence containing serine–glycine, found in mammalian aggrecans were observed in salmon, however, random serine–glycine (or glycine–serine) sequences predicted to the sugar chain attachment sites were observed. Based on cDNA analysis and amino acid analysis after β-elimination, the ratio of serine attached to sugar chains was calculated to be approximately 37.7% of total serine, that is, 46 of 123 serine residues.     

Optimization of butanol production from tropical maize stalk juice by fermentation with Clostridium beijerinckii NCIMB 8052

Mixed sugars from tropical maize stalk juice were used to carry out butanol fermentation with Clostridium beijerinckii NCIMB 8052. Batch experiments employing central composite design (CCD) and response surface methodology (RSM) optimization were performed to evaluate effects of three factors, i.e. pH, initial total sugar concentration, and agitation rate on butanol production. Optimum conditions of pH 6.7, sugar concentration 42.2 g/L and agitation rate 48 rpm were predicted, under which a maximum butanol yield of 0.27 g/g-sugar was estimated. Further experiments demonstrated that higher agitation facilitated acetone production, leading to lower butanol selectivity in total acetone–butanol–ethanol (ABE). While glucose and fructose are more preferable by C. beijerinckii, sucrose can also be easily degraded by the microorganism. This study indicated that RSM is a useful approach for optimizing operational conditions for butanol production, and demonstrated that tropical maize, with high yield of biomass and stalk sugars, is a promising biofuel crop.     

Pregnancy and conception rate after two intravaginal inseminations with dog semen frozen either with 5% glycerol or 5% ethylene glycol

The primary goal of this study was to compare the effects of 5% ethylene glycol (EG) and 5% glycerol (G) on fertility of frozen–thawed dog semen following intravaginal insemination. The sperm-rich fraction of the ejaculate of three male dogs was collected, pooled and divided into two aliquots, and then frozen with a Tris–glucose–egg yolk–citric acid extender containing either 5% G or 5% EG. A total of 10 bitches were inseminated twice, five with G-frozen–thawed semen and five with EG-frozen–thawed semen; intravaginal inseminations were performed the 4th and the 5th day after the estimated LH peak; four straws, thawed in a 37 °C water bath for 1 min and diluted in a Tris buffer, were used for insemination (200 × 10⁶ spermatozoa); the insemination dose was introduced in the cranial vagina of the bitch using a sterile plastic catheter. Ovariohysterectomy was performed in all bitches between days 29 and 31 after the calculated LH surge, and pregnancy status, and the number of conceptuses and corpora lutea were recorded. All bitches were pregnant. Neither the number of conceptuses, nor the ratio of conceptuses to corpora lutea (conception rate) was significantly different between groups. In this first screening, with a limited number of bitches, EG-frozen semen did not show a higher fertility than G-frozen semen when used for two intravaginal inseminations. Irrespective of the semen used, conception rate was 0.50.     

Alkaline conditions in hydrophilic interaction liquid chromatography for intracellular metabolite quantification using tandem mass spectrometry

Modeling of metabolic networks as part of systems metabolic engineering requires reliable quantitative experimental data of intracellular concentrations. The hydrophilic interaction liquid chromatography–electrospray ionization–tandem mass spectrometry (HILIC–ESI–MS/MS) method was used for quantitative profiling of more than 50 hydrophilic key metabolites of cellular metabolism. Without prior derivatization, sugar phosphates, organic acids, nucleotides, and amino acids were measured under alkaline and acidic mobile phase conditions with pre-optimized multiple reaction monitoring (MRM) transitions. Irrespective of the polarity mode of the acquisition method used, alkaline conditions achieved the best quantification limits and linear dynamic ranges. Fully 90% of the analyzed metabolites presented detection limits better than 0.5 pmol (on column), and 70% presented 1.5-fold higher signal intensities under alkaline mobile phase conditions. The quality of the method was further demonstrated by absolute quantification of selected metabolites in intracellular extracts of Escherichia coli. In addition, quantification bias caused by matrix effects was investigated by comparison of calibration strategies: standard-based external calibration, isotope dilution, and standard addition with internal standards. Here, we recommend the use of alkaline mobile phase with polymer-based zwitterionic hydrophilic interaction chromatography (ZIC–pHILIC) as the most sensitive scenario for absolute quantification for a broad range of metabolites.