In vitro evaluation of antibacterial activities and anti-inflammatory effects of Bifidobacterium spp. addressing acne vulgaris.

The objective of this study was to evaluate the antibacterial and anti-inflammatory effect of Bifidobacterium spp. In the first part of the study, the antibacterial activities of live and sonicated cells, from a total of 23 Bifidobacterium species, on the growth of 5 different strain of Staphylococcus aureus. Six strains, of sonicated Bifidobacterium, exhibited antibacterial activity against staphylococci samples, and seven Bifidobacterium strains exhibited antibacterial activity on the growth of S. aureus S.P.-N2. In the second part of the study, we tested the antimicrobial activity, of Bifidobacterium against Propionibacterium acne KCTC3320, using the co-culture method. The loss of P. acnes viability, caused by B. adolescentis SPM0308 and B. longum SPM1207, was 84% and 75%, respectively (*p < 0.05). In the third part of the study, the anti-inflammatory activity of B. adolescentis SPM0308 and B. longum SPM1207 were assessed; nitric oxide (NO), and tumor necrosis factor-α (TNF-α), production were tested using the murine macrophage RAW 264.7 cell line. Treatment of RAW 264.7 cells, with Bifidobacterium, decreased production of NO and TNF-α rather than LPS (100 ng/mL) treatment. The results suggest that B. adolescentis SPM0308 could be used as an effective control for P. acnes KCTC3320, and S. aureus, and reduce the risk of acne vulgaris development. We suggest that B. adolescentis SPM0308 may be a useful probiotic microorganism, for prevention of acne vulgaris, without adverse effects.

Increase of 30K protein in identified motoneurons by hemolymph results in inhibition of programmed cell death in silkworm, Bombyx mori (Lepidoptera, Bombycidae).

This study demonstrates that a 30K protein was gradually synthesized in primary-cultured motoneurons from the accessory planta retractor (APR) of the 6th abdominal ganglion (APR6) in silkworm ventral ganglia through stimulation of hemolymph. An increase in 30K protein synthesis resulted in an inhibition of programmed cell death (PCD) of APR6 motoneurons. The 30K protein was gradually synthesized from the 30Kc6 gene of identified APR6s in day-6 4th instars to day-9 5th instar larvae, but synthesis of the 30K protein ceased in isolated APR6s of day-1 pupa, which normally begin to undergo PCD. When pupal APR6s were treated with larval hemolymph, however, the 30K protein was synthesized suggesting the existence of an anti-PCD factor in the larval hemolymph. An increase of 30K protein within the APR6s was confirmed by antiserum made against the recombinant 30K protein that originated from the APR 30Kc6 gene. Larval APR6, in which PCD was induced with 20-hydroxyecdysone (20E) added to the primary culture, exhibited some PCD characteristics of shrinkage of cell bodies, axonal fragmentation and loss of mitochondrial function. These results provide new insights on the survival or PCD of insect motoneurons through stimulation of hemolymph.

Brain-derived neurotrophic factor promotes neurite growth and survival of antennal lobe neurons in brain from the silk moth, Bombyx mori in vitro.

This study was conducted to investigate effects of brain-derived neurotrophic factor on the neurite growth and the survival rate of antennal lobe neurons in vitro, and secretion of brain-derived neurotrophic factor-like neuropeptide from brain into hemolymph in the silk moth, Bombyx mori. In primary culture of antennal lobe neurons with brain-derived neurotrophic factor, it promoted both a neurite extension of putative antennal lobe projection neurons and an outgrowth of branches from principal neurites of putative antennal interneurons with significance (p<0.05). Brain-derived neurotrophic factor also increased significantly a survival rate of antennal lobe neurons (p<0.05). Results from immunolabeling of brain and retrocerebral complex, and ELISA assay of hemolymph showed that brain-derived neurotrophic factor-like neuropeptide was synthesized by both median and lateral neurosecretory cells of brain, then transported to corpora allata for storage, and finally secreted into hemolymph for action. These results will provide valuable information for differentiation of invertebrate brain neurons with brain-derived neurotrophic factor.

Effects of 20-hydroxyecdysone and serotonin on neurite growth and survival rate of antennal lobe neurons in pupal stage of the silk moth Bombyx mori in vitro.

Effects of 20-hydroxyecdysone and serotonin on the morphological development and the survival of antennal lobe neurons from day-2 pupal brains of the silk moth Bombyx mori were investigated in vitro. Four morphologically distinct neuronal types could be identified in the cultured antennal lobe neurons: unipolar, bipolar, multi-polar and projection neurons. Antennal lobe neurons in culture with 20-hydroxyecdysone and serotonin showed different patterns of the morphological development from those described in Manduca sexta. Projection neurons extend their neurites remarkably by 20-hydroxyecdysone in B. mori, but there is no extension from antennal lobe neurons in M. sexta. Multi-polar neurons conspicuously increase only formation of new branches from their primary neurites by serotonin in B. mori, but there are both extension and branching of the neurites in M. sexta. On day-5, antennal lobe neurons in lower titers of 20-hydroxyecdysone had significantly higher survival rates than those in higher titers. Neurons cultured for 7 days at different levels of 20-hydroxyecdysone generally showed significantly lower survival rates than neurons cultured for 5 days under the same conditions.

Molecular characterization of a cDNA from the silk moth Bombyx mori encoding Manduca sexta allatotropin peptide.

Allatotropin is a 13-residue amidated neuropeptide isolated from pharate adult heads of the tobacco hornworm, Manduca sexta and strongly stimulates biosynthesis of juvenile hormones in adults, but not larval, lepidopteran corpora allata. From a Bombyx mori midgut cDNA library, a cDNA that encodes a 130-amino-acid polypeptide containing M. sexta allatotropin sequence was isolated. The B. mori allatotropin cDNA consists of 1196 nucleotides. The encoded allatotropin peptide is identical to that isolated from M. sexta and that predicted from Pseudaletia unipuncta, with 84% and 81% identity in the amino acid sequence of the allatotropin peptide precursor, respectively. M. sexta allatotropin is flanked by two different endoproteolytic cleavage sites within the precursor of the B. mori allatotropin peptide. Evidence from northern blotting of B. mori tissues showed that the allatotropin gene is expressed in the cells of midgut, head and integument with different transcription amount, but not in the fat body and silk gland. Midgut has also a number of allatotropin-immunoreactive cells and nerve fibers. These results will provide valuable information in understanding the AT gene of insects.