Novel attacin from Hermetia illucens: cDNA cloning, characterization, and antibacterial properties.

In this study, we report a novel member of the attacin family from Hermetia illucens. The cDNA clone encoding the attacin-like protein was isolated by screening a cDNA library prepared from immunized fat body. The complete 510 bp cDNA of HI-attacin was predicted to encode a protein of 169 amino acids with a molecular weight of 17.7 kDa. The putative mature protein of H. illucens attacin (HI-attacin) had 50% identity with that of Bactrocera dorsalis attacin B. Phylogenetic analysis revealed that the HI-attacin was separated from the other dipteran attacins with a high bootstrap percentage. Compared to that in the other dipteran attacins, the G1 domain of HI-attacin was shorter and the sequence of the G2 domain of HI-attacin was more conserved than that of the G1 domain. We produced the recombinant attacin (rHI-attacin) protein using a prokaryotic expression system to confirm its antibacterial character. The rHI-attacin was produced as inclusion bodies and refolded by on-column refolding. rHI-attacin exhibited antibacterial activity against both Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA). Using real-time PCR, the expression of HI-attacin was was barely detected before the immunization, but was mostly evident in the fat body after immunization.

Outcome of Limited Video-Assisted Lumbar Sympathetic Block for Plantar Hyperhidrosis Using Clipping Method.

BACKGROUND: There are many ways to treat focal hyperhidrosis, including surgeries for palmar and axillary hyperhidrosis. However, doctors and patients tend to be reluctant to perform surgery for plantar hyperhidrosis due to misconceptions and prejudices about surgical treatment. In addition, few studies have reported the outcome of surgeries for plantar hyperhidrosis. Therefore, the objective of this study was to determine the outcome (early and late postoperative satisfaction, complication, compensatory hyperhidrosis, recurrence rate, and efficiency) of surgical treatment for plantar hyperhidrosis. MATERIALS AND METHODS: From August 2014 to October 2015, lumbar sympathetic block (LSB) was performed in 82 patients with plantar hyperhidrosis using clipping method. Limited video-assisted LSB was performed using 5 mm ligamax-clip or 3 mm horizontal-clip after identifying L3-4 sympathetic ganglion through finger-touch and endoscopic vision. RESULTS: Of the 82 patients, 45 were male and 37 were female. Their mean age was 26.38 years (range, 14-51 years). Mean follow-up time was 6.60 ± 3.56 months. Mean early postoperative satisfaction score was 9.6 on the 10th day postoperative evaluation. At more than 1 month later, the mean late postoperative satisfaction score was 9.2. There was no significant difference in early postoperative satisfaction score between clipping level L3 and L4/5. However, late postoperative satisfaction score was significantly better in the L3 group than that in the L4/5 group. Patient's age and body mass index did not affect the satisfaction score. However, male patients and patients who had history of hyperhidrosis operation showed higher satisfaction score than others. CONCLUSION: Limited video-assisted LSB using clip provided good results with minimal complications and low compensatory hidrosis, contrary to the prejudice toward it. Therefore, surgical treatment is recommended for plantar hyperhidrosis.

Purification and characterization of a novel antibacterial peptide from black soldier fly (Hermetia illucens) larvae.

In this study, we induced and purified a novel antimicrobial peptide exhibiting activity against Gram-positive bacteria from the immunized hemolymph of Hermetia illucens larvae. The immunized hemolymph was extracted, and the novel defensin-like peptide 4 (DLP4) was purified using solid-phase extraction and reverse-phase chromatography. The purified DLP4 demonstrated a molecular weight of 4267 Da, as determined using the matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) method. From analysis of DLP4 by N-terminal amino acid sequencing using Edman degradation, combined with MALDI-TOF and rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR), the amino acid sequence of the mature peptide was determined to be ATCDLLSPFKVGHAACAAHCIARGKRGGWCDKRAVCNCRK. In NCBI BLAST, the amino acid sequence of DPL4 was found to be 75% identical to the Phlebotomus duboscqi defensin. Analysis of the minimal inhibitory concentration (MIC) revealed that DLP4 have antibacterial effects against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA). The expression of DLP4 transcripts in several tissues after bacterial challenge was measured by quantitative real-time PCR. Expression of the DLP4 gene hardly occurred throughout the body before immunization, but was mostly evident in the fat body after immunization.