RESUMO Como elemento traço essencial para os animais, o cobre contribui significativamente para o crescimento e saúde dos animais. Comparado aos oligoelementos inorgânicos, os oligoelementos orgânicos são melhores suplementos; notavelmente, eles são adquiridos através de transformação microbiana. Portanto, nós selecionamos microorganismos enriquecidos com cobre de solos com alto teor de cobre para obter cobre orgânico. O carbamato de sódio diethyldithio trihidratado foi aplicado como agente cromogênico para a determinação de micro quantidades de cobre intracelular através da espectrofotometria. No total, 50 fungos foram isolados após a aplicação bem sucedida da plataforma de triagem para micróbios ricos em cobre. Após análises morfológicas e de biologia molecular, a cepa N-2, identificada como Aspergillus niger sp. demonstrou um melhor potencial de enriquecimento de cobre do que outras. Notavelmente, a tolerância da estirpe ao cobre foi quase três vezes maior que a da Saccharomyces cerevisiae, até 1600mg/L. O conteúdo de cobre ligado orgânico era de 22,84mg Cu/g de célula seca. Usando o método de superfície de resposta Central Composite Design (CCD), nós otimizamos a condição de fermentação (quantidade de inoculação, 13%; temperatura, 28C; pH, 5,0). Em comparação com os resultados da deformação original sob a condição de fermentação de fator único, relatamos um aumento de 24,18% sob as condições otimizadas. Coletivamente, estas descobertas fornecem uma referência para descobrir novos aditivos de cobre orgânico de baixo custo.

ABSTRACT As an essential trace element for animals, copper significantly contributes to the growth and health of animals. Compared to inorganic trace elements, organic trace elements are better supplements; notably, they are acquired through microbial transformation. Therefore, we screened for copper-enriched microorganisms from high copper content soil to obtain organic copper. Sodium diethyldithio carbamate trihydrate was applied as a chromogenic agent for determining micro amounts of intracellular copper through spectrophotometry. In total, 50 fungi were isolated after the successful application of the screening platform for copper-rich microbes. Following morphological and molecular biology analyses, the N-2 strain, identified as Aspergillus niger sp. demonstrated showed better copper enrichment potential than others. Notably, the strain tolerance to copper was nearly thrice that of Saccharomyces cerevisiae, up to 1600mg/L. The content of the organic bound copper was 22.84mg Cu/g dry cell. Using the Central Composite Design (CCD) response surface method, we optimized the fermentation condition (inoculation amount, 13%; temperature, 28(C; pH, 5.0). Compared to the original strain results under the single factor fermentation condition, we reported an increase by 24.18% under the optimized conditions. Collectively, these findings provide a reference for uncovering new and low-cost organic copper additives.

Abstract Plasma nitriding is a plasma-activated thermochemical method widely used to increase the fatigue strength, hardness and wear resistance of low-alloy steels. In this work, a new structure consisting of a hollow cathode metal plate can produce a hollow cathode discharge and heat the workpiece by radiation. This principle is used to perform hollow cathode source plasma nitriding (HCSPN). Herein, 42CrMo steels were treated by conventional plasma nitriding (CPN) and HCSPN treatments. The 42CrMo steels were nitrided at 525 °C for 6 h in NH3 atmosphere. The mechanical and dry wear properties were measured by nano-indentation, Vickers micro-indentation and ball-on-disk tribometer, respectively. The H3/E⁎2 ratio was 0.0274 GPa (H=11.51 GPa and E⁎=235.84 GPa) for the CPN sample and 0.0276 GPa (H=10.87 GPa and E⁎=215.54 GPa) for the HCSPN sample. Compared with the untreated 42CrMn steel, all the nitrided samples possess increased fracture toughness. Because the workpiece was not used as a discharge cathode, this HCSPN treatment overcame the disadvantages of the conventional CPN treatment. Additionally, results showed that the surface of the HCSPN sample was smoother than that of the CPN sample, and its tribological performance was better.

The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology.

Hydraulic support instability (HSI) is one of the most common causes of disasters in underground coal mining, posing a threat to the safety of mine workers and normal operation of the equipment. It is prone to occur in fully-mechanized mining faces with a large mining height (FMMLMH), especially when the dip angle of the coal seam is large. The key to controlling HSI is to deduce its mechanism and employ effective control techniques. This paper focuses on the analysis of HSI types, the key parameters and techniques to control HSI in FMMLMH, the establishment of a model of HSI in FMMLMH, and a multi-parameter sensitivity mechanical model of different HSI forms in the no. 7(2)19 longwall face in Xutuan Coal Mine, Huaibei Mining Group, by using sensitivity analysis . The results show that HSI mainly presents in three forms: hydraulic support gliding (HSG), hydraulic support tilting (HST), and hydraulic support tail twisting (HSTT). The occurrence of the above three forms depends mainly on support anti-instability capability. In the no. 7219 longwall face, HSG and HST are the main two forms of HSI. The dip angle of the working face and the friction coefficient between floor and hydraulic support are the sensitive parameters for HSG, while HST is strongly dependent on the dip angle of the working face and the friction coefficient between roof and hydraulic support. By the applications of measures such as the oblique layout of the working face, cutting the floor into a step pattern, moving the support under pressure, and raising the setting load, the support stability was controlled effectively.

Most cells exchange ions and small metabolites via gap junction channels. These channels are made of two hemichannels (connexons), each formed by the radial arrangement of six connexin (Cx) proteins. Connexins span the bilayer four times (M1-M4) and have both amino- and carboxy-termini (NT, CT) at the cytoplasmic side of the membrane, forming two extracellular loops (E1, E2) and one inner (IL) loop. The channels are regulated by gates that close with cytosolic acidification (e.g., CO2 treatment) or increased calcium concentration, possibly via calmodulin activation. Although gap junction regulation is still unclear, connexin domains involved in gating are being defined. We have recently focused on the CO2 gating sensitivity of Cx32, Cx38 and various mutants and chimeras expressed in Xenopus oocytes and studied by double voltage clamp. Cx32 is weakly sensitive to CO2, whereas Cx38 is highly sensitive. A Cx32 chimera containing the second half of the inner loop (IL2) of Cx38 was as sensitive to CO2 as Cx38, indicating that this domain plays an important role. Deletion of CT by 84% did not affect CO2 sensitivity, but replacement of 5 arginines (R) with sparagines (N) at the beginning of CT (C1) greatly enhanced the CO2 sensitivity of Cx32. This suggests that whereas most of CT is irrelevant, positive charges of C1 maintain the CO2 sensitivity of Cx32 low. As a hypothesis we have proposed a model that involves charge interaction between negative residues of the beginning of IL1 and positive residues of either C1 or IL2. Open and closed channels would result from IL1-C1 and IL1-IL2 interactions, respectively