Enhanced degradation of lignin by the strain fusion of Pseudomonas putida and Gordonia sp

The protoplast fusion technique was applied to construct a more efficient engineering microbial strain to degrade lignin by fusing two strains, Pseudomonas putida and Gordonia sp. At an initial lignin concentration of 900?mg/L, COD, BOD, TOC removal efficiencies increased from 69–76%, 69–72%, and 70–72% by the parent stains to 83%, 83%, and 83% of the fused strain, respectively. IR and HPLC analyses of the treated solution suggested that the fused strains were more capable of breaking the C_α–C_β bonds of the benzene ring in lignin compared to its parent strains, yielding syringyls as the main product. GC–MS analysis was used to identify the release of three-types of lower molecular intermediates: ring-opening, monomer, and dipolymer products. The phenolic hydroxyl group in lignin was oxidized to carbonyls, followed by further degradation to acids and esters. The carboxyl group on the ether linkage that maintains the macromolecular structure of lignin was oxidized to acyls, which further led to depolymerization and the opening of benzene ring.     

Sensitivity of mass spectrometry analysis depends on the shape of the filtration unit used for filter aided sample preparation (FASP)

Efficient protein solubilization using detergents is required for in‐depth proteome analysis, but successful LC‐MS/MS analysis greatly depends on proper detergents removal. A commonly used sample processing method is the filter‐aided sample preparation (FASP), which allows protein digestion and detergent removal on the same filtration device. Many optimizations of the FASP protocol have been published, but there is no information on the influence of the filtration unit typology on the detergents removal. The aim of this study was to compare the performance of conic and flat bottom filtration units in terms of number of proteins identified by LC‐MS/MS. We have analyzed 1, 10 and 100 μg of total cell lysate prepared using lysis buffer with different SDS concentrations. We compared the FASP protocol using conic and flat bottom filtration units to ethanol precipitation method. Subsequently, we applied our most performant protocol to single murine pancreatic islet, and identified up to 2463 protein using FASP versus 1169 proteins using ethanol precipitation. We conclude that FASP performance depends strongly on the filter shape: flat bottom devices are better suited for low‐protein samples, as they allow better SDS removal leading to the identification of greater number of proteins.     

1株滩涂沉积物反硝化细菌的鉴定及其性能研究

利用富集培养法从江苏沿海滩涂沉积物中分离筛选获得1株具有反硝化性能的细菌,命名为MD5。通过形态学观察、糖发酵及16S r DNA序列分析对其进行了分类鉴定、系统发育地位及反硝化性能等方面的研究。结果表明,该菌株能在厌氧条件下利用硝酸钠进行反硝化反应,在初始硝酸盐质量浓度为1 500 mg/L、30℃培养条件下,144 h后培养基中硝酸盐脱除率可达90.1%;在好氧培养条件下,该菌株可使能力测试管中硝酸盐质量浓度下降89.3%。菌株MD5的形态学、糖发酵结果与盐单胞菌一致,且基因序列与盐单胞属(Halomonas)细菌Halomonas sp.IW11-1(AB305262.1)的序列同源性达到99%,表明菌株MD5为1株盐单胞属细菌。本研究分离得到的菌株MD5在厌氧和好氧条件下均具有较强的反硝化能力,对海洋沉积物脱氮具有一定的意义。     

4种不同生活型湿地植物对富营养化水体的净化效果

选择4种湿地植物菖蒲、香蒲、浮萍和金鱼藻,研究单一及组合湿地植物对高浓度污水(污水处理厂进水)、低浓度污水(污水处理厂出水)中营养物质的去除效果.结果表明: 水体中总氮(TN)、总磷(TP)、化学需氧量(COD)浓度呈现试验前期快速下降,后期缓慢下降的趋势,表明湿地植物能有效净化污水中的TN、TP、COD,但不同湿地植物及湿地植物组合的净化效果存在差异.多种湿地植物组合比单种湿地植物对TN的净化作用强,其中香蒲＋浮萍＋金鱼藻对TN的净化效果最佳;高浓度污水中,单种挺水植物对TP的净化效果较好,低浓度污水中,则是多种湿地植物组合对TP的去除率较高;高浓度污水中,湿地植物对COD的去除率为85.1%～96.0%,其中菖蒲、香蒲去除效果最佳,低浓度污水中,湿地植物对COD去除率为76.9%～94.8%,以菖蒲＋浮萍＋金鱼藻去除效果最好.总体看来,湿地植物对高浓度污水中TN、TP、COD的净化效果好于低浓度污水,两种水体的pH都得到改善.     

Potential of aquatic macrophytes and artificial floating island for removing contaminants

Eutrophication is a major environmental issue that mankind is facing today as a result of rapid development. To reduce the eutrophication problem, we are testing an innovative artificial floating islands (AFIs) approach. AFIs include aquatic plants composed of emergent and floating macrophytes. In this experimental approach, two aquatic plants were selected to compare with the control group in order to evaluate the capacity of AFIs. These two emergent macrophytes were evaluated for a 40-day timeframe to examine their ability to purify waste water. The results showed that an AFI with purple loosestrife (Lythrum salicaria Linn.) and one with yellow-flowered iris (Iris wilsonii) have a strong ability to remove nitrogen, phosphorus, and other pollutants from water bodies. The pollutant removal rates of the AFIs with purple loosestrife and yellow-flowered iris were almost over 50%. The chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP) removal rates of the AFI with purple loosestrife were about 75%, 57%, 71%, respectively. At the same time, the COD, TN, and TP removal rates of the AFI with yellow-flowered iris were 60%, 49%, and 68%, respectively. AFIs with aeration are also a good way to remove pollutants. This study reveals that AFIs can be incorporated into wetlands to reduce the effects of water contamination and help strengthen wetland restoration.     

One-step partial or complete caries removal and bonding with antibacterial or traditional self-etch adhesives: study protocol for a randomized controlled trial

BackgroundCurrent concepts in conservative dentistry advocate minimally invasive dentistry and pulp vitality preservation. Moreover, complete removal of carious dentin in deep carious lesions often leads to pulp exposure and root canal treatment, despite the absence of irreversible pulp inflammation. For years, partial caries removal has been performed on primary teeth, but little evidence supports its effectiveness for permanent teeth. Furthermore, the recent development of new antibacterial adhesive systems could be interesting in the treatment of such lesions. The objectives of this study are to compare the effectiveness of partial versus complete carious dentin removal in deep lesions (primary objective) and the use of an antibacterial versus a traditional two-step self-etch adhesive system (main secondary objective).Methods/DesignThe DEep CAries Treatment (DECAT) study protocol is a multicenter, randomized, controlled superiority trial comparing partial versus complete caries removal followed by adhesive restoration. The minimum sample size required is 464 patients. Two successive randomizations will be performed (allocation ratio 1:1): the first for the type of excavation (partial versus complete) and the second (if no root canal treatment is required) for the type of adhesive (antibacterial versus traditional). For the two objectives, the outcome is the success of the treatment after 1 year, measured according to a composite outcome of five FDI criteria: material fracture and retention, marginal adaptation, radiographic examination (including apical pathologies), postoperative sensitivity and tooth vitality, and carious lesion recurrence.DiscussionThe study will investigate the interest of a conservative approach for the management of deep carious lesions in terms of dentin excavation and bioactive adhesive systems. The results may help practitioners achieve the most efficient restorative procedure to maintain pulp vitality and increase the restoration longevity.

Trial registration

ClinicalTrials.gov Identifier {"type":"clinical-trial","attrs":{"text":"NCT02286388","term_id":"NCT02286388"}}NCT02286388. Registered in November 2014.

Electronic supplementary material

The online version of this article (doi:10.1186/s13063-016-1484-0) contains supplementary material, which is available to authorized users.     

Morphological and physiological changes exhibited by a Cd-resistant Dictyosphaerium chlorelloides strain and its cadmium removal capacity

Changes induced on freshwater microalga Dictyosphaerium chlorelloides (Dc^wt) acclimated in the laboratory until their survival in culture media enriched with cadmium 100 µM have been studied. Cadmium removal by living cells of this Cd-resistant (Dc^CdR100) strain was tested in cultures exposed to 100 µM Cd during 30 days. Cell dimensions were measured under light microscopy, and cell growth was studied. Photosynthetic yield (Φ_PSII) was analyzed and the photosynthetic oxygen development and respiration response was obtained. Results show that Dc^CdR100 strain exhibited significant cell morphology changes in comparison to Dc^wt cells, which affected both surface area and cell biovolume. Malthusian fitness analysis showed that Dc^CdR100 strain living in Cd-enriched culture had developed a lower capacity of nearly 50% growth, and its photosynthetic oxygen development and respiration response were significantly reduced in both light and dark photosynthetic phases. Dc^CdR100 strain showed a very high capacity to remove cadmium from the aquatic environment (over 90%), although most of the removed heavy metal (≈70%) is adhered to the cell wall. These specific characteristics of Dc^CdR100 cells suggest the possibility of using this strain in conjunction with Dc^wt strain as bioelements into a dual-head biosensor, and in bioremediation processes on freshwater polluted with Cd.     

Current technologies to endotoxin detection and removal for biopharmaceutical purification

Endotoxins are the major contributors to the pyrogenic response caused by contaminated pharmaceutical products, formulation ingredients, and medical devices. Recombinant biopharmaceutical products are manufactured using living organisms, including Gram-negative bacteria. Upon the death of a Gram-negative bacterium, endotoxins (also known as lipopolysaccharides) in the outer cell membrane are released into the lysate where they can interact with and form bonds with biomolecules, including target therapeutic compounds. Endotoxin contamination of biologic products may also occur through water, raw materials such as excipients, media, additives, sera, equipment, containers closure systems, and expression systems used in manufacturing. The manufacturing process is, therefore, in critical need of methods to reduce and remove endotoxins by monitoring raw materials and in-process intermediates at critical steps, in addition to final drug product release testing. This review paper highlights a discussion on three major topics about endotoxin detection techniques, upstream processes for the production of therapeutic molecules, and downstream processes to eliminate endotoxins during product purification. Finally, we have evaluated the effectiveness of endotoxin removal processes from a perspective of high purity and low cost.     

Asymmetric competition between two tit species: a reciprocal removal experiment

1. In a 5-year field experiment, competition for food was tested between great tit ( Parus major L.) and blue tit ( P. caeruleus L.), two common hole-nesters in Central Europe. Experimental ('allopatric') populations of both species were created during the breeding seasons by preventing the nesting or egg laying of one of the competing species.
2. An asymmetric relationship was found between the two tits; blue tits were more successful in the competitive interaction. Detectable effects were found only in nestling condition. Great tits raised lighter nestlings when breeding sympatrically with blue tits.
3. A possible mechanism is suggested that could be responsible for the different competitive abilities of the two species; blue tits are more effective in utilizing the most abundant size categories of caterpillar food supply than great tits.     

Immunohistochemical visualization of insulin receptors in formalin-fixed bovine ovaries post mortem and in granulosa cells collected in vivo

Insulin is crucial for granulosa cell (GC) function, follicle growth and ovulation in cows; low insulin levels increase the risk for anoestrus. Apart from insulin concentration, alterations in the insulin receptor (IR) density on GC may affect follicular growth and steroidogenesis. Data about the IR protein distribution in the bovine follicle are scarce. Therefore, we aimed to develop a quantifiable staining method for IR protein on histological sections of bovine follicles in different developmental stages, and to apply this technique on GC obtained in living cows.In a first experiment, bovine ovaries were collected post mortem, formalin fixed, routinely processed, and stained with monoclonal murine IR-antibodies, peroxidase-labeled goat anti-mouse antibodies, and substrate chromogen. Based on their diameter, follicles were morphologically classified as small antral (SAF; n = 141), dominant (DF; n = 28) or subordinate (SF; n = 8); DF and SF were further classified as healthy or atretic based on the ratio of estrogen and progesterone concentrations in their follicular fluid. Using specialized software, the proportion of pixels displaying a positive staining signal was computed as a measure for IR density in three selected follicular regions: GC, theca (T) and stroma (STR). Results were analyzed in an ANOVA model with follicle type, region and health status as fixed factors. In SAF, DF, and SF, IR density was notably higher in GC than T or STR; the latter two displayed very low or no IR presence. The IR density in SAF was stronger than in DF and tended to be stronger than in SF. Staining intensity was not altered in atretic compared to healthy follicles. In corpus luteum, cumulus-oocyte complexes and pre-antral follicles, no IR could be detected. In a second experiment, GC samples were collected from 20 live cows on 30 and 70 d post partum by transvaginal follicular fluid aspiration, projected on glass slides, and stained using the protocol described above. Most samples yielded sufficient GC and IR was clearly visualized. However, objective quantification of the staining signal was impeded by extensive variation in the arrangement and density of GC and the amount of cellular debris on the slides.Altogether, strong IR presence in GC, most notably in SAF, suggests acquisition of IR as a key event in early follicle growth. Furthermore, we have developed a quantifiable staining technique for bovine follicles that may be applicable for GC obtained in live cows, although this method requires further standardization.