The 30K proteins are involved with important functions in the growth and development of Bombyx mori. In this study, the synthesis and regulation of 30K proteins were examined during the degeneration of Bombyx silk glands. On day 3 of the fifth instar, the protein level of 30Kc19 was low, whereas the silk proteins were rapidly synthesized. However, synthesis and accumulation of the 30Kc19 protein significantly increased at the prepupal stage and on day 1 of the pupal stage. At this stage, the silk gland cells were filled with 30Kc19 and genomic DNA. Moreover, the transcript levels of the 30K-encoding genes, including 30Kc6, 30Kc12, 30Kc19 and 30Kc23 were up-regulated during the degeneration of the Bombyx silk glands. During the time that the levels of the 30Kc19 protein were significantly up-regulated, it is notable that the transcript levels of the BmAtg8, BmAtg6 and BmDronc genes dramatically increased to regulate the programmed cell death of this gland. On day 1 of the pupal stage, intense fragmentation of genomic DNA occurred in the silk gland cells, and the putative active form of caspase was detected in the cytoplasm, showing the complete degradation of the silk glands in one day. In conclusion, the 30K proteins are synthesized in high concentrations, while proteolysis mediates silk gland degeneration in Bombyx by a caspase-dependent pathway. We propose that the 30K proteins may be nutrients and energy vectors to be absorbed by the developing tissues of pupae or moths.
Single-step real-time high-throughput monitoring of drug influences on bacterial cell behaviour has become important with growing interests in personalized therapy and medication. Conventional microchip assemblies to perform similar work do exist. However, most of these devices have complex set-ups incorporating micromixers, separators, pumps, or valves. These microcomponents can sometimes damage the entities being monitored because of the creation of unfavourable biological environments. This paper presents a microchip-based system that enables single-step mixing of two solutions in various ratios, without the need for additional microcomponents such as mixers and pumps, in order to screen effectively their combinatory effects on cell outcomes. In this work, in-vitro experiments were carried out using ampicillin at various concentrations to investigate their effects on Escherichia coli (E. coli). Results showed that the microchip provided effective screening, which yielded useful results such as effective dosages, ineffective dosages, and other possible outcomes; for instance, in this case, the occurrence of adaptive mutation of the bacteria at certain drug concentrations. Comparative microbiological laboratory tests were carried out as standard for confirmation of the results.
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