Upload this completed form to website with submission ARTICLE INFORMATION Fill in information in each box below Article Type Research article Article Title Investigation of Immunostimulatory Effects of Heat-treated Lactiplantibacillus plantarum LM1004 and its Underlying Molecular Mechanisms Running Title (within 10 words) Immunostimulatory Effects of Heat-treated Lactiplantibacillus plantarum LM1004 Author Won-

PurposeAcne vulgaris is a common skin disease accompanied by chronic inflammation in the pilosebaceous follicles, resulting from excessive Cutibacterium acnes. This study aimed to investigate the inhibition of biofilm formation by C. acnes ATCC 6919 using heat‐treated Pediococcus acidilactici LM1013 (HT‐LM1013), previously isolated from the Korean traditional fermented alcoholic beverage—makgeolli, and its application as a leave‐on‐type product for patients with acne vulgaris.MethodsHT‐LM1013 was prepared by Lactomason and homogenized using a high‐pressure homogenizer. The minimum inhibitory concentration (MIC), tricarboxylic acid (TCA) cycle, and lipase activity were evaluated for C. acnes inhibition. Inhibition of biofilm formation was demonstrated using a crystal violet solution. Damaged C. acnes was observed using field‐emission scanning electron microscopy (FE‐SEM). Clinical trials were performed using a leave‐on‐type product containing HT‐LM1013.ResultsHT‐LM1013 inhibited the TCA cycle (36.80%) and lipase activity using palmitate (31.89%), stearate (36.91%), and oleate (30.86%) as substrates at 1 × MIC (p < 0.01). After treatment with HT‐LM1013, concave and elongated shapes of C. acnes were observed by FE‐SEM. In addition, HT‐LM1013 inhibited biofilm formation by 71.75% at 1 × MIC (p < 0.001) and removed 73.35% of mature biofilms (p < 0.01). In the clinical trial, the leave‐on‐type product decreased the number of closed comedones from 14.04 to 10.22, open comedones from 7.22 to 4.39%, and sebum content to 76.23% at week 4 (p < 0.01). The satisfaction score of the participants was recorded 3.83 on a five‐point scale.ConclusionHT‐LM1013 is potent for the treatment of acne vulgaris.

Probiotics are defined as live organisms in the host that contribute to health benefits. Lactobacillus gasseri LM1065, isolated from human breast milk, was investigated for its probiotic properties based on its genome. Human breast milk was donated by a healthy woman. The isolated Lactobacillus strain was identified as L. gasseri and named LM1065. Complete genome sequencing and de novo assembly were performed using the PacBio RS II system and hierarchical genome assembly process (HGAP). Probiotic properties were determined by the resistance of the strain to gastric conditions, adherence ability, enzyme production, and safety assessment. The fungistatic effect and the ability to inhibit hyphae transition were studied using the cell-free supernatant (CFS) of L. gasseri LM1065. L. gasseri LM1065 showed high gastric pepsin tolerance and mild tolerance to bile salts. Auto-aggregation ability and hydrophobicity were measured to be 61.21% and 61.55%, respectively. These properties resulted in the adherence of the bacteria to the human intestinal epithelial cells (2.02%). In addition, antibiotic-resistance genes and putative virulence genes were not predicted in the complete genome sequence of L. gasseri LM1065, and antibiotic susceptibility was satisfied by the criteria of the European Food Safety Authority using a cut-off test. CFS showed a fungistatic effect and suppressed the tricarboxylic acid cycle in Candida albicans (29.02%). CFS also inhibited the transition to true hyphae and damaged the blastoconidia. This study demonstrates the essential properties of this novel probiotic, L. gasseri LM1065, and its potential to inhibit vaginal Candida albicans infection.

Probiotics are defined as live organisms in the host that contribute to health benefits. Lactobacillus gasseri LM1065, isolated from human breast milk, was investigated for its probiotic properties based on its genome. Draft genome map and de novo assembly were performed using the PacBio RS II system and hierarchical genome assembly process (HGAP). Probiotic properties were determined by the resistance to gastric conditions, adherence ability, enzyme production, safety assessment and mobile genetic elements. The fungistatic effect and inhibition of hyphae transition were studied using the cell-free supernatant (CFS). L. gasseri LM1065 showed high gastric pepsin tolerance and mild tolerance to bile salts. Auto-aggregation and hydrophobicity were measured to be 61.21% and 61.55%, respectively. The adherence to the human intestinal epithelial cells was measured to be 2.02%. Antibiotic-resistance genes and putative virulence genes were not predicted in the genomic analysis, and antibiotic susceptibility was satisfied by the criteria of the European Food Safety Authority. CFS showed a fungistatic effect and suppressed the tricarboxylic acid cycle in Candida albicans (29.02%). CFS also inhibited the transition to true hyphae and damaged the blastoconidia. This study demonstrates the essential properties of this novel probiotic, L. gasseri LM1065, and potential to inhibit vaginal C. albicans infection.