Jack bean α-mannosidase digestion profile of hybrid-type N-glycans: effect of reaction pH on substrate preference

Jack bean α-mannosidase (JBM) is a well-studied plant vacuolar α-mannosidase, and is widely used as a tool for the enzymatic analysis of sugar chains of glycoproteins. In this study, the JBM digestion profile of hybrid-type N-glycans was examined using pyridylamino (PA-) sugar chains. The digestion efficiencies of the PA-labeled hybrid-type N-glycans Manα1,6(Manα1,3)Manα1,6(GlcNAcβ1,2Manα1,3)Manβ1,4GlcNAcβ1,4GlcNAc-PA (GNM5-PA) and Manα1,6(Manα1,3)Manα1,6(Galβ1,4GlcNAcβ1,2Manα1,3)Manβ1,4GlcNAcβ1,4GlcNAc-PA (GalGNM5-PA) were significantly lower than that of the oligomannose-type N-glycan Manα1,6(Manα1,3)Manα1,6Manβ1,4GlcNAcβ1,4GlcNAc-PA (M4-PA), and the trimming pathways of GNM5-PA and GalGNM5-PA were different from that of M4-PA, suggesting a steric hindrance to the JBM activity caused by GlcNAcβ1-2Man(α) residues of the hybrid-type N-glycans. We also found that the substrate preference of JBM for the terminal Manα1-6Man(α) and Manα1-3Man(α) linkages in the hybrid-type N-glycans was altered by the change in reaction pH, suggesting a pH-dependent change in the enzyme-substrate interaction.     

Evaluation of velvet bean meal as an alternative protein ingredient for poultry feed

The effect of certain simple and cost-effective processing methods on the nutritional and anti-nutritional properties of seed materials of an under-utilized food legume, Mucuna pruriens (L.) DC. var. utilis (Wall. ex Wight) Baker ex Burck (velvet bean, VB), collected from Valanadu, Kerala, India was analyzed in experiment 1. The raw VB seeds were found to contain appreciable levels of crude protein (263.2 g/kg dry matter (DM)); ether extract (79.6 g/kg DM); crude fiber (95.8 g/kg DM) and ash content (38.4 g/kg DM). Among the different treatments used, soaking in sodium bicarbonate solution + autoclaving was more effective in reducing maximum levels of various anti-nutritional compounds of VB seeds. Furthermore, in experiment 2, the effect of inclusion of different levels of velvet bean meal (VBM; subjected to soaking in sodium bicarbonate solution + autoclaving) as an alternative protein source in poultry feed on the growth performance of commercial-type broiler birds was investigated. The results indicate that the inclusion of VBM up to the 40% level exhibited better growth performance of the broiler birds such as feed intake, body weight gain, feed conversion ratio and protein efficiency ratio in both the starter and finisher phases.     

Evaluation of acrylamide grafted moth bean starch as controlled release excipient

The aim of this study was to investigate the applicability of acrylamide grafted moth bean starch as controlled release matrix former. Lamivudine was used as model drug and its controlled release tablets were formulated using various concentration of grafted copolymer. The grafted copolymer was tested for acute toxicity and drug-excipient compatibility study. The formulations were evaluated for physical characteristics like hardness, friability, % drug content and weight variations. The in vitro release study showed that the optimized formulation exhibited highest correlation (R) value in case of Higuchi model and the release mechanism study proved that the formulation showed a combination of diffusion and erosion based release process. There was a significant difference in the pharmacokinetic parameters (T_max, C_max, AUC, V_d, T_1/2 and MDT) of the optimized formulation as compared to the marketed conventional tablet Lamivir^®, which confirmed controlled release potential of acrylamide grafted copolymer.     

Multivesicular bodies in developing tobacco seed and mung bean are functionally equivalent

Protein storage vacuoles (PSVs) are the primarily storage organelles in cotyledon cells for protein preservation in seeds. Storage proteins are transported from the endoplasmic reticulum (ER) to the Golgi apparatus for subsequent delivery to PSVs via presumably Golgi-derived dense vesicles (DVs). However, recent studies demonstrated that storage proteins in early stage of developing cotyledon of mung beans reached the multivesicular bodies (MVBs) prior to the detection of DVs, indicating the possible involvement of MVBs in mediating transport of storage proteins during the early stage of seed development. Here, we further show that the MVBs in developing tobacco seeds are functionally and biochemically equivalent to those in developing mung beans. Thus, MVBs in developing tobacco seeds are structurally distinct from DVs, contain both vacuolar sorting receptors (VSRs) and storage proteins, and they are insensitive to treatments of wortmannin and brefeldin A (BFA).     

低频电刺激+耳穴压豆+中药汤药治疗宫内残留物的效果及对超声指标的影响

摘要 目的：探讨低频电刺激+耳穴压豆+中药汤药治疗宫内残留物的效果及对超声指标的影响。方法：选择2022年1月至12月来我院诊治的存在宫内残留物患者120例,根据随机数字表法,将120例患者分为A组、B组、C组、D组,每组30例,A组患者采用生化汤加减治疗,B组使用低频电刺激+生化汤加减治疗,C组使用耳穴压豆+生化汤加减治疗,D组低频电刺激+耳穴压豆+生化汤加减治疗。对比四组治疗前后的宫内残留物超声影像表现,对比四组的超声RI、PI水平、中医证候积分、凝血功能指标,对比四组的血β-HCG水平恢复正常时间、月经复潮时间、阴道流血持续时间、临床疗效及不良反应发生情况。结果：治疗前,四组患者均有宫内残留物,声像的图像表现为子宫局灶性增厚,子宫内膜的回声欠均匀,厚薄不均,宫腔内局部可见不均质的回声团,边界欠清晰;治疗后,四组超声检查发现部分患者无宫内残留物;部分患者的宫内残留物回声明显减低。治疗前,四组宫内残留物超声RI、PI水平、中医证候积分、凝血功能指标对比无差异（P>0.05）;治疗后,四组的RI、PI水平较治疗前明显升高,B、C、D组明显较A组高,D组明显较B、C组高（P均<0.05）;四组的阴道出血量、小腹痛、大便干、舌苔、脉象评分、全血低切粘度、全血高切粘度、纤维蛋白原较治疗前明显降低,B、C、D组明显较A组低,D组明显较B、C组低（P均<0.05）。B、C、D组的血β-HCG水平恢复正常时间、月经复潮时间、阴道流血持续时间明显较A组低,D组的明显较B、C组低（P均<0.05）;B、C组以上指标对比无差异（P>0.05）。B、C、D组的临床疗效较A组高,D组较B、C组高,四组患者的临床疗效对比无差异（P>0.05）。治疗过程中,四组均无明显不良反应,完成治疗。结论：低频电刺激+耳穴压豆+生化汤加减治疗可明显提高血瘀证宫内残留物的效果,可能与其可改善患者的凝血功能有关,阴道彩色多普勒超声可用于评估宫内残留物治疗的疗效。     

Water stress and insect herbivory interactively reduce crop yield while the insect pollination benefit is conserved

Climate change is predicted to hamper crop production due to precipitation deficits and warmer temperatures inducing both water stress and increasing herbivory due to more abundant insect pests. Consequently, crop yields will be impacted simultaneously by abiotic and biotic stressors. Extensive yield losses due to such climate change stressors might, however, be mitigated by ecosystem services such as insect pollination. We examined the single and combined effects of water stress, insect herbivory and insect pollination on faba bean yield components and above‐ and belowground plant biomass under realistic field conditions. We used rainout shelters to simulate a scenario in line with climate change projections, with adequate water supply at sowing followed by a long period without precipitation. This induced a gradually increasing water stress, culminating around crop flowering and yield formation. We found that gradually increasing water stress combined with insect herbivory by aphids interactively shaped yield in faba beans. Individually, aphid herbivory reduced yield by 79% and water stress reduced yield by 52%. However, the combined effect of water stress and aphid herbivory reduced yield less (84%) than the sum of the individual stressor effects. In contrast, insect pollination increased yield by 68% independently of water availability and insect herbivory. Our results suggest that yield losses can be greatly reduced when both water stress and insect herbivory are reduced simultaneously. In contrast, reducing only one stressor has negligible benefits on yield as long as the crop is suffering from the other stressor. We call for further exploration of interactions among ecosystem services and biotic and abiotic stressors that simulate realistic conditions under climate change.     

Conjugated and free sterols from black bean (Phaseolus vulgaris L.) seed coats as cholesterol micelle disruptors and their effect on lipid metabolism and cholesterol transport in rat primary hepatocytes

Phytosterols have been widely studied for their cholesterol-lowering effect. Conjugated phytosterol forms have been found more active than free moieties. There are no reports about the sterol profile of black bean seed coats neither its effects on cholesterol metabolism. The aim of this research was to identify and quantify phytosterols from black bean seed coats and to determine their effects on cholesterol micellar solubility and on mRNA and key protein levels involved in lipid/cholesterol metabolism and cholesterol transport in primary rat hepatocytes. Free phytosterols, acylated steryl glycosides, and steryl glycosides were extracted from black bean seed coats. They were identified through HPLC–MS–TOF and quantified through HPLC equipped with UV–visible and evaporative light-scattering detectors. Free and conjugated phytosterols from the coats significantly increased the inhibitory effect of cholesterol micelle formation compared with stigmasterol, which was used as control (P < 0.05). In addition, phytosterols of black bean seed coat decreased lipogenesis by the downregulation of lipogenic proteins such as sterol regulatory element-binding protein 1 and fatty acid synthesis (FAS) in primary rat hepatocytes. Regarding β-oxidation, phytosterols upregulated the expression of carnitine palmitoyltransferase I and promoted the β-oxidation of long-chain fatty acids. Phytosterols inhibited cholesterol micellar solubility and reduced the activation of the liver X receptor, decreasing hepatic FAS and promoting hepatic β-oxidation of long-chain fatty acids.     

Adaptive Changes and Genotypic Variation for Root Architecture of Common Bean in Response to Phosphorus Deficiency

Root architectural responses to phosphorus (p) availability may be an important trait for P acquisition efficiency. In the present study, The authors examined the effects of P availability on root architectural responses of different common bean genotypes. Five common bean (Phaseolus vulgaris L.) genotypes representing different origins and ecotypic races were compared both in a specially designed paper pouch system and a stratified P buffer sand culture system with computer image analysis. The results showed that root architecture was regulated by P availability. P deficiency led to form a shallower root system, as indicated by increased relative distribution of basal root length in the upper layers and decreased the growth angle of basal roots. There was significant genetic variation in root architecture in response to P deficiency both in the paper pouch system and the stratified sand culture system. Under low P conditions some genotypes were more gravitropically sensitive to low P availability, resulting in producing a shallower root system and enhanced root exploration into the surface soil, where soil available P is more concentrated. G19833 and DOR364, which were most contrasting in P efficiency, were also very different in root architectural response to P availability. The results from this study suggest that P availability regulates root architecture and P deficiency leads to shallower root architecture in beans. The genetic potential of root architecture provides the possibility of selecting this trait for improving P acquisition efficiency in common bean.     

Osmotic regulation of assimilate unloading from seed coats of Vicia faba. Assimilate partitioning to and within attached seed coats.

The significance of the osmotic potential of the seed apoplast sap as a regulator of assimilate transfer to and within coats of developing seed of Vicia faba (cv. Coles Prolific) was assessed using attached empty seed coats and intact developing seed. Following surgical removal of the embryos, through windows cut in the pod walls and underlying seed coats, the resulting attached “empty” seed coats were filled with solutions of known osmotic potentials (–0. 02 versus –0. 75 MPa). Sucrose efflux from the coats was elevated at the higher osmotic potential (high osmotic concentration) for the first 190 min of exchange. Thereafter, this efflux was depressed relative to efflux from coats exposed to the low osmotic potential (high osmotic concentration) solution. This subsequent reversal in efflux was attributable to an enhanced diminution of the coat sucrose pools at the high external osmotic potential. Indeed, when expressed as a proportion of the current sucrose pool size, relative efflux remained elevated for coats exposed to the high osmotic potential solution. Measurement of potassium and sucrose fluxes to and from their respective pools in the coat tissues demonstrated that the principal, fluxes, sensitive to variative in the external osmotic potential, were phloem import into and efflux from the “empty” coats. Phloem import, consistent with a pressure-driven phloem transport mechanism, responded inversely with changes in the external osmotic potential. In contrast, sucrose and potassium efflux from the coats exhibited a positive dependence on the osmotic potential. Growth rates of whole seed were approximately doubled by enclosing selected pods in water jackets held at temperatures of 25°C. compared to 15°C. The osmotic potential of sap collected from the seed apoplast remained constant and independent of the temperature-induced changes in seed growth rates and hence phloem import. Based on these findings, it is proposed that control of phloem import by changes in the external osmotic potential observed with “empty” seed coats has no significance as a regulator of assimilate import by intact seed. Rather, maintenance of the seed apoplast osmotic potential, independent of seed growth rate, suggests that the observed osmotic regulation of efflux from the coats may play a key role in integrating assimilate demand by the embryo with phloem import.