Chondrogenic differentiation of rat mesenchymal stem cells on silk fibroin/chondroitin sulfate/hyaluronic acid ternary scaffolds

Cartilage repair is a challenge in bone tissue reconstruction. In this study, silk fibroin (SF), chondroitin sulfate (CS) and hyaluronic acid (HA) were employed to fabricate scaffolds for tissue engineered cartilage by freeze drying technique. The secondary pores were formed in the main pores of SF/CS/HA scaffold which improved the pore connectivity and equilibrium swelling of the scaffold. Furthermore, rat bone marrow mesenchymal stem cells were seeded on the scaffolds to evaluate the cell adhesion and proliferation. Results of hematoxylin/eosin staining and cell counting kit-8 assay showed that the cells migration and differentiation of SF/CS/HA (80/15/5) scaffold were better than that of SF/CS/HA scaffolds with different ratios after 7 days culture. Moreover, immunohistochemistry and scanning electron microscope demonstrated that large amounts of collagen II and proteoglycans of the cells were expressed in the SF/CS/HA 3D scaffold, while the expression of collagen I was barely visible by immunohistochemistry. Abound of extracellular matrix was formed to morphologically round and distributed uniformly throughout the scaffolds. The 3D ternary scaffold could promote the cells chondrogenic differentiation without using any inductive agent and offer potential for cartilage tissue regeneration.     

The effect of growth regulators on tuberisation of cultured stem pieces ofSolanum Tuberosum

Summary The effect of growth regulators on tuberisation of potato (var.British Queen) stem cuttings has been investigated. It was found that indoleacetic acid and maleic hydrazide promoted tuberisation while gibberellic acid strongly inhibited the formation of tubers. The behaviour of stem pieces on a low sugar medium was also investigated and it was found that the application of growth regulators did not cause tuberisation when an adequate supply of carbohydrate was not available. It is suggested that tuberisation is regulated by the interplay of carbohydrate supply, gibberellins and antigibberellin-like substances.     

Fabrication of hollow nanofibrous structures using a triple layering method for vascular scaffold applications

This study aims to develop a new approach for fabricating hollow nanofibrous yarns by engineering a triple-layer structure (polyvinyl alcohol (PVA) multifilament core surrounded by a layer of PVA nanofibers and a polylactic acid (PLA) nanofiber outer layer). After fabrication of this 3-layer structure, the core portion was extracted, leaving the outer layer intact after dissolving the PVA nanofibers in water. To determine the optimum thickness of the outer layer, hollow nanofiber yarns with five different thicknesses were produced. A hollow nanofiber yarn was also produced using a common method to enable comparison of the methods. In the common method, a core sheath yarn consisting of a PVA multifilament core and a PLA nanofiber outer layer was fabricated, and a hollow yarn was produced by placing the core yarn in hot water. The results revealed facilitation of core extraction from the yarn body of the new 3-layer structure, which occurred due to rapid dissolution of the middle layer. The wicking behavior in the hollow yarn fabricated using the novel method followed the Locus Washburn equation and that of the hollow yarn produced from the core sheath yarn deviated from it. The results demonstrated that tensile properties of hollow nanofiber yarns were improved by increasing the thickness. Furthermore, hemolysis and cytotoxicity assays indicated that the fabricated hollow nanofibrous structure is non-toxic and blood compatible, indicating its potential for use in biomedical applications such as vascular scaffolds.     

播娘蒿高频率再生植株因素的研究

以播娘蒿子叶、子叶柄和下胚轴为外植体，研究不同外植体、激素组合以及硝酸银对播娘蒿再生频率的影响。结果表明：外植体、激素以及硝酸银的影响都非常显著；最佳外植体为下胚轴；最佳激素组合为6－BA（2．0mg/L) NAA(0.5mg/L);附加AgNO3(1.7mg/L)可使下胚轴再生频率提高63．42％。诱导生根的最佳培养基为1／2MS＋NAA（0．5mg/L) IBA(0.5mg/L)。     

Impact of vitronectin concentration and surface properties on the stable propagation of human embryonic stem cells

The standard method for culturing human embryonic stem cells (hESC) uses supporting feeder layers of cells or an undefined substrate, Matrigel(?), which is a basement membrane extracted from murine sarcoma. For stem cell therapeutic applications, a superior alternative would be a defined, artificial surface that is based on immobilized human plasma vitronectin (VN), which is an adhesion-mediating protein. Therefore, VN adsorbed to diverse polymer surfaces was explored for the continuous propagation of hESC. Cells propagated on VN-coated tissue culture polystyrene (TCPS) are karyotypically normal after >10 passages of continuous culture, and are able to differentiate into embryoid bodies containing all three germ layers. Expansion rates and pluripotent marker expression verified that a minimal VN surface density threshold is required on TCPS. Further exploration of adsorbed VN was conducted on polymer substrates with different properties, ranging from hydrophilic to hydrophobic and including cationic and anionic polyelectrolyte coatings. Despite differing surface properties, these substrates adsorbed VN above the required surface density threshold and were capable of supporting hESC expansion for >10 passages. Correlating wettability of the VN-coated surfaces with the response of cultured hESC, higher cell expansion rates and OCT-4 expression levels were found for VN-coated TCPS, which exhibits a water contact angle close to 65°. Importantly, this simple, defined surface matches the performance of the benchmark Matrigel, which is a hydrogel with highly complex composition.     

Distribution and effectiveness of pesticide application with a cold fogger on pepper plants cultured in a greenhouse

A cold fogger was evaluated for efficiency of spray-deposit distribution and for control of insect damage and fungal disease on pepper (Capsicum annuum L.) plants cultured in a greenhouse. The fogger was placed at one end of an 800-m² greenhouse, and the distribution and deposition of a fluorescent tracer by the cold fogger was evaluated using quantitative methods and image analysis. An increased cold fogger-dependent air speed, determined by anemometry, was correlated with increased deposition of the fluorescent tracer. A consistent decline of deposition from the area nearest to the cold fogger to the far end of the greenhouse was observed. Nonetheless, no differences were found in the droplet density between the outer and inner parts of the foliage. The average droplet density was 1828 cm⁻², but the leaf deposition was restricted to the upper surface of the leaves. The distribution and numbers of thrips (Thrips sp.) and powdery mildew (Leveillula taurica [Lev.] G. Arnaud) on leaves were assessed over a period of five months. Thrips control was found to be quite good throughout the greenhouse in response to phytosanitary treatments. Although the control of powdery mildew was generally adequate, there was significant disease development in certain locations in the greenhouse that were distant from the cold fogger. The employment of cold foggers in greenhouses needs to be carefully analysed in terms of the dimensions of the greenhouse and the pests and diseases to be controlled due to the uneven deposition of the sprayed products.     

Adhesion, proliferation, and gene expression profile of human umbilical vein endothelial cells cultured on bilayered polyelectrolyte coatings composed of glycosaminoglycans

This study characterized human umbilical vein endothelial cell (HUVEC) adhesion, proliferation, and gene expression on bilayered polyelectrolyte coatings composed of an outermost layer of glycosaminoglycans (hyaluronan, heparin, or chondroitin sulfate), with an underlying layer of poly-L-lysine or chitosan. The proportion of cells that adhered to the various polyelectrolyte coatings after 1 and 2 h incubations was quantified by the WST-8 assay. Interchanging poly-L-lysine with chitosan resulted in significant differences in cellular adhesion to the outermost glycosaminoglycan layer after 1 h, but these differences became insignificant after 2 h. The proliferation of HUVEC on the various bilayered polyelectrolyte coatings over 10 days was characterized using the WST-8 assay. Regardless of whether the underlying layer was poly-L-lysine or chitosan, HUVEC proliferation on the hyaluronan outermost layer was significantly less than on heparin or chondroitin sulfate. Additionally, it was observed that there was more proliferation with poly-L-lysine as the underlying layer, compared to chitosan. Subsequently, real-time polymerase chain reaction was used to analyze the expression of seven genes related to adhesion, migration, and endothelial function (VWF, VEGFR, VEGFA, endoglin, integrin-α5, ICAM1, and ICAM2) by HUVEC cultured on the various bilayered polyelectrolyte coatings for 3 days. With poly-L-lysine as the underlying layer, biologically significant differences (greater than twofold) in the expression of VWF, VEGFR, VEGFA, endoglin, and ICAM1 were observed among the three glycosaminoglycans. With chitosan as the underlying layer, all three glycosaminoglycans displayed biologically significant differences in the expression of VWF and VEGFR compared to the chitosan control. CT-HA displayed the highest level of expression of VWF, whereas expression levels of VEGFR were almost similar among the three glycosaminoglycans.     

Determination of drift potential of different flat fan nozzles on a boom sprayer using a test bench

This study's objective was to evaluate the functionality of an ad hoc test bench for spray drift measurement with boom sprayers, using it for evaluating different nozzles according to drift risk. The repeatability of results was evaluated by conducting similar tests at two different laboratories. Drift potential values (DPV) obtained showed an interesting effect of Venturi flat fan nozzles on drift reduction, in comparison with conventional flat fan nozzles (reference nozzle was XR 11003). Newly designed flat fan nozzles reduced the risk of drift. Reasonably relations between 10th-percentile, D[v,0.1], 50th-percentile or Volume Median Diameter, D[v,0.5], 90th-percentile, D[v,0.9], V₁₀₀ and DPV were observed in all cases, with R² values of 0.58, 0.65, 0.66 and 0.72, respectively. The lowest drift values were achieved with TTI and TD Spray Max nozzles; they were significantly lower than those obtained for IDK and AIXR ones. Results indicated that the drift test bench can be used as an alternative to the official standard procedure for drift measurements on boom sprayers (e.g. ISO 22866), as it is able to discriminate the influence of different boom settings (especially nozzle types) on drift. Further studies could be useful in order to prove that the classification of nozzles according to drift risk obtained using the test bench is comparable to the nozzle classifications obtained applying the ISO 22866 test method.     

The effect of stem population on tuber yield in a trial with single-stem seed pieces

Summary Excised eyes on 5 g of seed tuber tissue were used to grow plants with single stems. These were transplanted in the field at densities of 120 000, 180 000 and 240 000 plants/ha, with in-row spacings of 16 or 33 cm. Total tuber yield was not affected, although more tubers were produced at the high densities. Tuber numbers did not increase in proportion to the number of stems. This was due to the fact that the number of tubers per stem decreased from a mean of 4.8 to 2.9 with increasing stem populations.     

The history and distribution of nodulating Paraburkholderia,a potential inoculum for Fynbos forage species

Legumes in the Fynbos vegetation of the Western Cape of South Africa have emerged as candidates for domestication, particularly for their adaptation to acidic and infertile soils. However, South African rhizobia have been shown to be very diverse and unique, and a detailed understanding of them is essential to success in forage breeding programs that seek to exploit these “new” legumes. Symbionts of legumes in South Africa that belong to traditional rhizobial genera have been shown to have a unique origin for their symbiotic loci in comparison to members sampled from other regions of the world. Some of the legume tribes in the Fynbos have also been shown to associate predominantly with unique species in the Betaproteobacterial genus Paraburkholderia. The rhizobial members of this genus have two main centres of diversity, of which South Africa is one. In this centre, the legume hosts are principally from the Papilionoideae subfamily while hosts from the mimosoid clade (now in the Caesalpinioideae) are abundant in the South American centre. Not only do these rhizobia differ in terms of host, but their symbiotic loci also show separate origins. The dominance and uniqueness of the Paraburkholderia symbionts, in the context of indigenous South African legumes, makes understanding the history and factors that affect the distribution of this genus essential if successful adaptation and effective nodulation of these legumes in Agriculture are to be achieved globally.     

甘蓝型油菜减氮增效潜力评价及种质资源筛选

为培育具有高潜力的减肥增效油菜品种,以不同甘蓝型油菜种质资源为材料,通过两年三点的田间肥效试验,研究了它们对氮肥的响应特征及节氮增产潜力.以产量平均值为分界线将供试材料划分为4种类型:低氮高效型、双高效型、双低效型和高氮高效型.在高氮条件下,双高效型和高氮高效型材料具有16.96％～36.50％的节氮潜力和21.57％...     

Methods of testing fungicides for controlling skin spot (Polyscytalum pustulans) on potatoes during storage

Summary Suspensions of 12 fungicides were evaluated for controlling skin spot on stored potatoes in two experiments. Tubers produced from seed tubers inoculated withPolyscytalum pustulans (Experiment 1) or tubers wounded with pin pricks before dipping in mycelial suspensions ofP. pustulans (Experiment 2) were immersed for 5 minutes in suspensions of different fungicides at 10 mg l. 100 mg 1 or 1 g l a.i. After incubation in sealed plastic bags at 5°C the numbers of skin spots developed at random over the tuber surface (Experiment 1) or at pin pricks (Experiment 2) were recorded after 20 or 24 weeks. In both experiments carbendazim, thiabendazole and thiophanate methyl and in one experiment thiophamine were most effective in decreasing incidence of skin spots. Imazalil, prochloraz and sisthane also had highly significant effects at higher concentrations (100mg l. l g l a. i).

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Zusammenfassung Es wurden 12 Fungizide auf ihre Wirkung auf das Myzelwachstum vonPolyscytalum pustulans und auf das Auftreten der Tüpfelfleckenkrankheit an gelagerten Kartoffelknollen geprüft. Formulierte Fungizide wurden in Konzentrationen von 0.3 bis 80 mg l in 2%igen, auf pH 7 abgepufferten Malzagar inkorporiert. Der durchmesser der Kolonien wurde nach 4-w?chiger Inkubation bei 15°C gemessen, die meisten Verbindungen ergaben niedrige Hemmkonzentrationen (IC₅₀, Tabelle 1). Die Prüfung der Fungizide erfolgte in 2 Versuchen durch 5-minütige Immersion von Knollen in Suspensionen von 10 mg 1, 100 mg 1 und 1 g l Wirkstoff. In Versuch 1 wurden die Knollen von Saatgut, welches zur Pflanzung mitP. pustulans inokuliert worden war. 5 Tage nach der Rodung behandelt. In Versuch 2 wurden 8 Wochen lang gelagerte Knollen gewaschen und dann mit einem Ger?t, mit 25 Nagelspitzen (1 mm lang, 0,5 mm Durchmesser an der Basis) in 3 mm Abstand in einer Platte (20 mm × 20 mm × 3 mm) mit Handgriff, verletzt (Abb. 1). Das Ger?t wurde an 2 Stellen in die Knollen gedrückt, die verletzten Knollen wurden unmittelbar danach in eine Suspension von 10 Wochen alten mazerierten Kulturen vonP. pustulans von 2%igem Malzagar getaucht. Am n?chsten Tag wurden die Knollen mit den Fungiziden behandelt. In beiden F?llen wurden die Knollen nach Abtropfen luftgetrocknet und bei 5°C in drahtverschlossenen Plastikbeuteln gelagert. Ermittelt wurde die Zahl der Tüpfelflecken (Abb. 1) bei zuf?lligem Auftreten (Versuch 1, 20 Wochen) und nach Verletzung mit Nagelspitzen (Versuch 2, 24 Wochen). Carbendazim, Thiabendazol, Thiophanatmethyl und Thiophamine waren die effektivsten Verbindungen zur Verhinderung von Tüpfelflecken an gelagerten Knollen (Tabellen 2 und 3). Imazalil, Prochloraz und Sisthane hatten bei h?heren Konzentrationen ebenfalls hochsignifikante Wirkung. Die Knollenbesch?digung verursacht neue Verwundungen, an denen sich Tüpfelflecken bilden k?nnen, deshalb sind Fungizidprüfungen offensichtlich auch zu anderen Zeitpunkten als unmittelbar nach der Rodung m?glich, wie dies bei natürlich infizierten Knollen erforderlich ist. Ausserdem war das Auftreten von Tüpfelflecken zwischen Proben besch?digter Knollen weniger variabel als bei Knollen aus inokuliertem Pflanzgut.

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Résumé Douze fongicides ont été testés afin d'observer leur action sur la croissance mycélienne dePolyscytalum pustulans et l'incidence de l'oosporiose sur les tubercules en conservation. Des formulations de fongicides ont été incorporées dans la gélose avec 2% d'extrait de malt tamponné à pH 7 à des concentrations variant de 0,3 à 80 mg/l. Le diamètre des colonies a été mesuré à 15°C après 4 semaines d'incubation. La plupart des produits ont donné une inhibition à des concentrations faibles (IC₅₀, tableau 1). Deux expérimentations ont été mises en place pour tester les fongicides après immersion des tubercules pendant 5 minutes dans des suspensions de 10 mg/l, 100 mg/l et 1 g/l de matière active. Dans la première expérimentation, les tubercules provenant de plants inoculés avecP. pustulans à la plantation ont ètè traités 5 jours après l'arrachage. Dans la deuxième expérience, les tubercules ont été conservés pendant 8 semaines, puis la vés et blessés à l'aide d'une plaque (fig. 1) en laiton (20 mm×20 mm×3 mm) munie de 25 pointes (1 mm de long 0,5 mm de large à la base) espacees de 3 mm chacune. Chaque tubercule est blessé de deux endroits et immergé immédiatement dans une suspension contenant une culture deP. pustulans agée de 10 semaines et macérée dans une solution d'extrait de malt à 2%. Les tubercules ont été traités avec les fongicides le jour suivant. Dans les deux expérimentations, les tubercules traités ont été ressuyés puis séchés et mis dans des sacs en plastique. Les sacs sont fermés et stockés à 5°CC. Le nombre de taches sur les tubercules, soit présentes sur toute la surface après 20 semaines dans la première expérimentation, soit aux endroits des blessures après 24 semaines dans la deuxième expérimentation, a été enregistré (fig. 1). Le carbendazime, le thiabendazole, le méthyl thiophanate et la thiophamine ont été les produits les plus efficaces vis à vis de l'oosporiose sur tubercules en conservation (tableaux 2 et 3). L'imazalil, le prochloraz et le sisthane ont eu des effets significatifs aux concentrations les plus élevées. Les tubercules blessés pour l'expérimentation provoquent des conditions favorables au développement de l'oosporiose; il est donc suggéré de tester les fongicides à d'autres dates que celle de l'arrachage où l'on utilise obligatoirement des tubercules infectés naturellement. De plus, en ce qui concerne l'expression des sympt?mes, les échantillons de tubercules blessés présent nt une variabilité moins importante que des tubercules provenant de semences inoculées.

     

Influence of Poly(lactic acid) Layer on the Physical and Antibacterial Properties of Dry Bacterial Cellulose Sheet for Potential Acute Wound Healing Materials

Dry bacterial cellulose nanofiber (BC) sheet coated with poly(lactic acid) (PLA) was developed and characterized towards acute wound healing applications. This new approach of PLA coating on BC revealed enhanced physical and antibacterial properties. Commercial BC sheets originated from the manufacturing of nata de coco jelly were dried and coated with the PLA at various concentrations of 2, 4, 6, 8, 10 and 12 % w/v for the purpose of improving the mechanical properties and followed by loading of antiseptic such as benzalkonium chloride (BAC). PLA has been proposed for the use of coating materials at a concentration of 8 %, the biocomposite sheet started exhibiting a low moisture uptake, prolonged swelling in simulated wound fluid solution and high tear (9.17 Nm²/kg) and burst indices (32.5 kPa·m²/g). The 8 % PLA coating revealed porous fiber-like morphology as observed under scanning electron microscope. Therapeutic loading capacity of the BC/8 PLA was substantially higher than the pristine BC. Furthermore strong antimicrobial activities against Staphylococcus aureaus and Escherichia coli were observed for the BC/8PLA biocomposite film. These reports were clearly suggestive of the fact that synthetic biodegradable polymers, such as PLA, may be exploited for the synergistic combination with BC for antimicrobial and acute wound management. This new and modified fiber source material could reduce the dependency on plant based cellulose for more demanding biomedical applications such as wound healing materials, vascular graft, cartilage replacement, drug delivery and tissue engineering.     

Lupins as a potential source of raffinose family oligosaccharides: Preparative method for their isolation and purification

Results of studies on the content of raffinose family oligosaccharides (RFOs) in plants of 11 species of the Leguminosae family are presented. It is clearly shown that plants of the Leguminosae family, especially of the lupin species, are the richest source of these sugars among the plants tested. A simple method of preparative isolation and purification of RFOs from bitter lupin extract was developed. This method seems to be useful for efficient isolation of oligosaccharides from plant material for further nutritional tests.     

果园生草模式土壤固碳潜力——以福建省为例

以定位试验为平台,研究了果园生草管理模式对土壤有机碳含量的影响,并通过分析土壤有机碳的变化量来估算福建省果园生草管理的土壤固碳潜力。结果表明：与1996年相比,2007年生草果园0-20 cm土壤有机碳含量提高了37.0%,土壤容重降低了1.8%。与清耕模式相比,生草模式更有利于土壤有机碳的积累,其土壤碳年变化量是清耕模式的3.2倍,土壤固碳速率为714.52 kg.hm-2.a-1。福建省果园生草模式土壤固碳潜力为3.26×105t.a-1,其中,以漳州地区最高,占36.3%。     

Feeding behavior of a potential insect pest, Lygus hesperus, on four new industrial crops for the arid southwestern USA

Camelina (Camelina sativa), guayule (Parthenium argentatum), lesquerella (Physaria fendleri), and vernonia (Centrapalus pauciflorus [formerly Vernonia galamensis]) are either under limited commercial production or being developed for production in the southwestern USA. Insect pests are a potential economic threat to all these new crops, with Lygus hesperus, the western tarnished plant bug, among the most prominent due to its regional abundance and propensity to feed on reproductive plant tissue. The objectives of this study were to establish baseline data on the feeding behavior and potential impact of L. hesperus on camelina, guayule, lesquerella and vernonia. Behavioral observations of adult females and males, and nymphs of this insect were made in the laboratory. Insects spent ≈35% of their time either probing (=tasting) or feeding on various reproductive and vegetative tissues of guayule, lesquerella or vernonia, but only 20% on camelina. When insects did probe and feed they preferred reproductive tissues, primarily flowers and siliques/achenes, and there were differences in these behaviors relative to crop but not generally to insect stage or sex. Insects probed and fed more on flower tissue of guayule and vernonia compared with camelina and lesquerella, and more on siliques of lesquerella compared with achenes of vernonia. When probing and feeding on vegetative tissue, there was generally a preference for stems compared with leaves in all crops except guayule. Results show that L. hesperus will readily feed on the economically important tissues of all crops, and although research has shown that this feeding did not consistently affect lesquerella yield, further work will be needed to determine if such feeding poses a risk to commercial production of camelina, guayule or vernonia.     

Determination of the potential land for securing double-rice cropping in the Cambodian Mekong Delta,based on a sub-area based modeling of flood inundation

Inundated areas of the Mekong Delta are the most important regions for agricultural production in both Cambodia and Vietnam. With population growth and increasing water demand in the dry season, effectively managing available water in the region is vital for crop production. This study is aimed at developing a sub-area based modeling of flood inundation model in order to analyze flood inundation processes in the Cambodian Mekong Delta as a basis for introducing semi-flood control for new cropping systems such as double-rice cropping. The simulated results of the flood inundation model from 2002 to 2003 were utilized for estimating land appearance in the dividing sub-area, and the potential of securing land for double-rice cropping was determined based on the period of land appearance and the cropping schedule. In order to realize the available water for double cropping, consideration of the effect of operating canal gates for controlling the early and receding inundation are crucially necessary. By considering the operation of control gates facilities, the potential land for securing double-rice cropping exceeded about 50 km² compared to the 34 km² of the actual land used for recession rice (about 30 and 20% of the total selected area, respectively). The study confirms that the sub-area based modeling of inundation model can be a helpful tool for water management in the Cambodia Mekong Delta.