Detection of recombinant human lactoferrin and lysozyme produced in a bitransgenic cow

Lactoferrin and lysozyme are 2 glycoproteins with great antimicrobial activity, being part of the nonspecific defensive system of human milk, though their use in commercial products is difficult because human milk is a limited source. Therefore, many investigations have been carried out to produce those proteins in biological systems, such as bacteria, yeasts, or plants. Mammals seem to be more suitable as expression systems for human proteins, however, especially for those that are glycosylated. In the present study, we developed a bicistronic commercial vector containing a goat β-casein promoter and an internal ribosome entry site fragment between the human lactoferrin and human lysozyme genes to allow the introduction of both genes into bovine adult fibroblasts in a single transfection. Embryos were obtained by somatic cell nuclear transfer, and, after 6 transferences to recipients, 3 pregnancies and 1 viable bitransgenic calf were obtained. The presence of the vector was confirmed by fluorescent in situ hybridization of skin cells. At 13 mo of life and after artificial induction of lactation, both recombinant proteins were found in the colostrum and milk of the bitransgenic calf. Human lactoferrin concentration in the colostrum was 0.0098 mg/mL and that in milk was 0.011 mg/mL; human lysozyme concentration in the colostrum was 0.0022 mg/mL and that in milk was 0.0024 mg/mL. The molar concentration of both human proteins revealed no differences in protein production of the internal ribosome entry site upstream and downstream protein. The enzymatic activity of lysozyme in the transgenic milk was comparable to that of human milk, being 6 and 10 times higher than that of bovine lysozyme present in milk. This work represents an important step to obtain multiple proteins or enhance single protein production by using animal pharming and fewer regulatory and antibiotic-resistant foreign sequences, allowing the design of humanized milk with added biological value for newborn nutrition and development. Transgenic animals can offer a unique opportunity to the dairy industry, providing starting materials suitable to develop specific products with high added value.     

Influence of low temperature prefermentative techniques on chromatic and phenolic characteristics of Syrah and Cabernet Sauvignon wines

Low temperature prefermentative techniques (cold maceration, superficially frozen grapes and dry-ice frozen must) were studied as an enological alternative for obtaining highly colored wines and compared with other more common practices, such as the use of maceration enzymes during the maceration step. The experiments were conducted with two grape varieties (Syrah and Cabernet Sauvignon) during the 2007 season. Significant differences were found between the treatments at the three moments analyzed (at the end of alcoholic and malolactic fermentation, and at the moment of bottling). The cold maceration technique produced the highest concentration of anthocyanins in Cabernet Sauvignon wines. As regards chromatic parameters, the prefermentative low temperature techniques led to wines with a better color than the control wine although similar to the color obtained when a commercial maceration enzyme was used. Taking into account the results in both Cabernet Sauvignon and Syrah wines, freezing the must with dry-ice and the use of maceration enzyme were the most interesting techniques.     

生产高质量气瓶钢时无缺陷板坯连铸工艺的最新进展

气瓶钢为裂纹敏感钢种,在Krakatan钢厂3号板坯连铸机上研究,减少板坯表面纵裂纹、横裂纹和星形裂纹,从而取消板坯清理,采用优化连铸工艺参数(如选择合适连铸保护渣、优化二冷冷却、液面控制、过热度、拉速等),研究其与板坯表面质量之间的关系.所选择的保护渣粘度和熔点低,冷却水强度从0.67L/kg钢,降低到0.58L/kg钢,拉速为1.1m/min,液面控制精度为±5%,钢水的含碳量0.09%左右.结果表明,横裂纹和纵裂纹减少90%,星形裂纹从62.21%减少到28.87%,板坯表面清理率从85.54%减少到25%以下,板坯修理损失率从3.4%降到0.05%.     

Generation of partially coherent diffraction-free fields with tunable geometry

The synthesis of diffraction-free fields with different profiles is described. The synthesis is done by illuminating a screen containing a circular slit with a cosine beam. The treatment is equivalent to the modulation of the slit transfer characteristics and makes possible a tunable interference interaction of Bessel beams with noncommon axes. These results are generalized, and it is shown that coherent diffraction-free fields with arbitrary profiles can be expressed as the superposition of shifted zero-order Bessel beams and temporary parametric representations of the shifted functions. Diffraction-free fields with partially coherent features can be obtained. Experimental results are shown for each case.     

New organic chemistry of sulfur dioxide

Simple 1,3-dienes undergo highly stereoselective hetero-Diels-Alder additions with SO2 at low temperature giving sultines. These reactions that are faster than the more exothermic cheletropic additions of SO2-producing sulfolenes. This has led to the invention of a new C-C bond-forming reaction combining electron-rich dienes and alkenes with SO2. The reaction cascade has been exploited to develop combinatorial, one-pot, four-component syntheses of polyfunctional sulfones, sulfonamides, and sulfonic esters. It also allows us to generate, in one-pot operations, enantiomerically enriched polypropionate fragments containing up to three contiguous stereogenic centers and a (E)-alkene unit. These fragments can be used directly in further C-C bond-forming reactions, such as cross-aldol condensations, thus permitting the expeditious construction of complicated natural products of biological interest (e.g., Baconipyrones, Rifamycin S, Apoptolidinone) and analogues. New ene reactions of SO2 have also been discovered; they open new avenues to organic synthesis.     

Occurrence and persistence of Listeria spp. in the environment of ewe and cow's milk cheese dairies in Portugal unveiled by an integrated analysis of identification, typing and spatial-temporal mapping along production cycle

Eight dairies, located in two distant geographic regions of Portugal, were screened along the production cycle in order to evaluate the presence and distribution of Listeria spp. in their environment. Three dairies in each region were positive for the presence of listeriae and 213 isolates were obtained. Based on an integrated analysis of RAPD fingerprints with three primers, molecular identification and genomic typing of isolates was performed followed by spatial and temporal mapping on dairy plants. The occurrence of Listeria species by region was noticeable different. Listeria monocytogenes prevailed in South Portugal dairies and L. innocua presented the highest occurrence in Azores, whereas L. seeligeri and L. ivanovii were detected in distinct regions. Dairies were at risk of contamination, from more than one source, whatever the stage in the production cycle and the surface materials used. For the three prevalent species, most of the genomic types were dairy and sampling time specific. Nonetheless, more than one type could be found in each dairy at a particular site and, in a few cases, even for different species. Some dairies also shared types, mainly for L. innocua and usually at the same stage of the production cycle. For L. monocytogenes, PCR serotyping was applied and 52% of genomic types were serotype 4b. An equal frequency of genomic types (24%) was found for serotypes 1/2b or 3b and 1/2a or 3a. The global pattern of types within a dairy is not constant, suggesting cycles of elimination and recontamination along the production cycle.     

Clarification of the slope mass rating parameters assisted by SMRTool,an open-source software

Bulletin of Engineering Geology and the Environment - Geomechanics classifications are used to perform a preliminary assessment of rock slope stability for different purposes in civil and mining...     

Energy‐efficient complex distillation sequences: Control properties

Four thermally coupled distillation systems were designed for the separation of five‐component mixtures (the light‐ends separation section of a crude distillation plant); their steady‐state design was obtained by starting from a conventional distillation sequence and then optimizing for minimum energy consumption. The thermally coupled distillation systems were compared to sequence based on conventional columns design. Comparison was based on controllability properties under open and closed loop operation, following the dynamic behaviour after common industrial operating disturbances. Simulation results were analyzed by the singular value decomposition technique and with the performance examination of elimination of feed disturbances using PI controllers. It was found that thermally coupled distillation systems are controllable and, sometimes, they exhibit dynamic responses that are easier to manage than in the case of conventional distillation sequences; this result is innovative in the study of this kind of systems.     

Control of Polymorphism and Morphology in Solution Sheared Organic Field‐Effect Transistors

During the last decades, small molecule organic semiconductors have been successfully used as active layer in organic field‐effect transistors (OFETs). Despite the high mobility achieved so far with organic molecules, in order to progress in the field it is crucial to find techniques to process them from solution. The device reproducibility is one of the principal weak points of organic electronics for further commercialization. To achieve a high device‐to‐device reproducibility it is essential to control the morphology and polymorphism of the active layer for OFET application. In this work, the preparation of thin films is reported based on blends of the organic semiconductor dibenzo‐tetrathiafulvalene (DB‐TTF) and polystyrene by a solution shearing technique compatible with upscaling. Here, it is demonstrated that varying the deposition parameters (i.e., speed and temperature) or the solution formulation (i.e., semiconductor/binder polymer ratio) is possible to control the film morphology and semiconductor polymorphism and, hence, the different intermolecular interactions. It is demonstrated that the control of the thermodynamics and kinetics of the crystallization process is key for the device performance optimization. Further, this is the first time that DB‐TTF thin films of the α‐polymorph are reported.     

On the stability of the troposphere/lower stratosphere and its relationships with cirrus clouds and three mandatory levels over Buenos Aires

An unrotated principal components analysis was carried out to establish the most representative modes for the joint variability between the heights of the upper and lower boundaries of cirrus clouds and three different mandatory levels (850, 500, and 100 hPa), and the associated stability of the troposphere over Buenos Aires. Discussion is limited to the first three most representative structures found, which consists of spatial patterns (or empirical orthogonal functions, EOFs) and their time-evolving coefficients (or principal components, PCs). EOF₁ shows a direct (indirect) mode that encompasses the cirrus slightly below (above) its mean position, with 500 and 100 hPa exhibiting a similar behaviour and 850 hPa acting the opposite way. EOF₁ is associated with above-normal stability (instability) for direct (indirect) modes (i.e. positive (negative) values of PC₁). On a monthly average, this occurs in the austral winter (summer) months. Regarding EOF₂, all three mandatory levels experience positive (negative) height anomalies in direct (indirect) modes and cirrus goes up (down) under mild stability (instability). Monthly averages show that PC₂ is approximately positive in summer and in early fall and negative the rest of the year. As to EOF₃, it is characterized by a stability similar to that of EOF₂, with direct (indirect) modes showing lowered (raised) cirrus and all three mandatory levels above (below) normal conditions; on a monthly basis, PC₃ is best described as having a semi-annual evolution, with maxima (minima) in March and October (January and August). Overall, EOF₁ has the highest stability or instability, depending upon the sign of PC₁. These results are the first of their kind worldwide.