Single-Cell Hemoprotein Diet Changes Adipose Tissue Distributions and Re-Shapes Gut Microbiota in High-Fat Diet-Induced Obese Mice.

We have previously observed that feeding with single-cell hemoprotein (heme-SCP) in dogs (1 g/day for 6 days) and broiler chickens (1 ppm for 32 days) increased the proportion of lactic acid bacteria in the gut while reducing their body weights by approximately 1~2%. To define the roles of heme-SCP in modulating body weight and gut microbiota, obese C57BL/6N mice were administered varied heme-SCP concentrations (0, 0.05, and 0.5% heme-SCP in high fat diet) for 28 days. The heme-SCP diet seemed to restrain weight gain till day 14, but the mice gained weight again later, showing no significant differences in weight. However, the heme-SCP-fed mice had stiffer and oilier bodies compared with those of the control mice, which had flabby bodies and dull coats. When mice were dissected at day 10, the obese mice fed with heme-SCP exhibited a reduction in subcutaneous fat with an increase in muscle mass. The effect of heme-SCP on the obesity-associated dyslipidemia tended to be corroborated by the blood parameters (triglyceride, total cholesterol, and C-reactive protein) at day 10, though the correlation was not clear at day 28. Notably, the heme-SCP diet altered gut microbiota, leading to the proliferation of known anti-obesity biomarkers such as Akkermansia, Alistipes, Oscillibacter, Ruminococcus, Roseburia, and Faecalibacterium. This study suggests the potential of heme-SCP as an anti-obesity supplement, which modulates serum biochemistry and gut microbiota in high-fat diet-induced obese mice.

Single-cell hemoprotein (heme-SCP) exerts the prebiotic potential to establish a healthy gut microbiota in small pet dogs.

To investigate the effect of the single-cell hemoprotein (heme-SCP) source on animals, a dog-treat (100 g for each dog) harboring 0.2% heme-SCP was manufactured and fed to seven pet dogs (< 10 kg) in a randomized manner (irrespective of owner's feeding style, dogs' health conditions, and staple diets), and the feces before and after the dog-treat diet were analyzed to define the structure of the microbiota. The total bacterial species of the seven dogs showed no difference (564-584), although the bacterial compositions varied significantly. The Firmicutes phylum increased (54.7-73.7%), showing differential species composition before and after heme-SCP intake. Proteobacteria, Bacteroidetes, and Fusobacteria decreased (5.4-3.8%, 32.9-16.8%, and 6.3-3.6%, respectively), which agreed with the previous observation of deliberate feeding. Therefore, it is conceivable that heme-SCP as a prebiotic can shape the gut microbiota regardless of the administration method. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01195-9.

Reverse Engineering Targets for Recombinant Protein Production in Corynebacterium glutamicum Inspired by a Fast-Growing Evolved Descendant.

We previously reported a Corynebacterium glutamicum JH41 strain with a 58% faster growth rate through application of adaptive laboratory evolution. To verify that the fast-reproducing strain was useful as a host for recombinant protein expression, we introduced a plasmid responsible for the secretory production of a recombinant protein. The JH41 strain harboring the plasmid indeed produced the secretory recombinant protein at a 2.7-fold greater rate than its ancestral strain. To provide the reverse engineering targets responsible for boosting recombinant protein production and cell reproduction, we compared the genome sequence of the JH41 strain with its ancestral strain. Among the 15 genomic variations, a point mutation was confirmed in the 14 bases upstream of NCgl1959 (encoding a presumed siderophore-binding protein). This mutation allowed derepression of NCgl1959, thereby increasing iron consumption and ATP generation. A point mutation in the structural gene ramA (A239G), a LuxR-type global transcription regulator involved in central metabolism, allowed an increase in glucose consumption. Therefore, mutations to increase the iron and carbon consumption were concluded as being responsible for the enhanced production of recombinant protein and cell reproduction in the evolved host.

Accelerated Growth of Corynebacterium glutamicum by Up-Regulating Stress- Responsive Genes Based on Transcriptome Analysis of a Fast-Doubling Evolved Strain.

Corynebacterium glutamicum, an important industrial strain, has a relatively slower reproduction rate. To acquire a growth-boosted C. glutamicum, a descendant strain was isolated from a continuous culture after 600 generations. The isolated descendant C. glutamicum, JH41 strain, was able to double 58% faster (td=1.15 h) than the parental type strain (PT, td=1.82 h). To understand the factors boosting reproduction, the transcriptomes of JH41 and PT strains were compared. The mRNAs involved in respiration and TCA cycle were upregulated. The intracellular ATP of the JH41 strain was 50% greater than the PT strain. The upregulation of NCgl1610 operon (a putative dyp-type heme peroxidase, a putative copper chaperone, and a putative copper importer) that presumed to role in the assembly and redox control of cytochrome c oxidase was found in the JH41 transcriptome. Plasmid-driven expression of the operon enabled the PT strain to double 19% faster (td=1.82 h) than its control (td=2.17 h) with 14% greater activity of cytochrome c oxidase and 27% greater intracellular ATP under the oxidative stress conditions. Upregulations of genes those might enhance translation fitness were also found in the JH41 transcriptome. Plasmid-driven expressions of NCgl0171 (encoding a cold-shock protein) and NCgl2435 (encoding a putative peptidyl-tRNA hydrolase) enabled the PT to double 22% and 32% faster than its control, respectively (empty vector: td=1.93 h, CspA: td=1.58 h, and Pth: td=1.44 h). Based on the results, the factors boosting growth rate in C. gluctamicum were further discussed in the viewpoints of cellular energy state, oxidative stress management, and translation.

A novel combination regimen with intense focused ultrasound and pressure- and dose-controlled transcutaneous pneumatic injection of hypertonic glucose solution for lifting and tightening of the aging face.

BACKGROUND: Intense focused ultrasound (IFUS) has demonstrated moderate efficacy for facial laxity of the aging face. Pressure- and dose-controlled transcutaneous pneumatic injections of hypertonic glucose solution (TPIG) are a minimally invasive way to deliver solution into the skin for therapeutic purposes. Recently, its application around temporal areas is known to exert early-onset lifting effects with facial contour rejuvenation. AIMS: We sought to evaluate the safety and efficacy of this novel combination regimen with IFUS and TPIG for lifting and tightening of aging face. PATIENTS AND METHODS: Twenty-two Korean subjects with mild-to-moderate facial skin laxity were evaluated after receiving a sequential single session of IFUS and TPIG treatments. Dermatologists' objective assessments for general appearance, and mid- and low faces based on photographic images were performed at 1- and 12-week post-treatment follow-ups. Patients' subjective assessments were also conducted. Skin biopsies were taken at baseline and the last visit. RESULTS: Among 22 subjects, twenty (91%) demonstrated improvement after 12-week follow-up. We also observed similarly improved state just after 1 week of treatments (82%: 18/22). Specifically, improvement of the mid-face was evident from the first week, while the progress of the low face improvement was observed at the twelve week. Subjective assessments paralleled these findings. No seriously adverse effect was observed during procedures. Histologic evaluation showed greater dermal collagen fibers throughout the dermis after treatments. CONCLUSION: Combination treatment with IFUS and TPIG has beneficial effects for skin lifting and tightening with early-onset time.

Real-time measurement of fibers using an HY-differential mobility analyzer with an optical particle counter (KOFAM).

This study investigated the applicability of an HY-differential mobility analyzer with an optical particle counter (HY-DMA/OPC), named as KOFAM, for counting fibrous matters in real time. Fibers separated from particles by the HY-DMA were counted with an OPC. To assess the KOFAM performance, the proposed method and the conventional gold standard phase contrast microscopy (PCM) method were compared in terms of variables such as recovery, relative difference, coefficient of determination, and conformity. The optimal sheath-to-aerosol (outlet) flow ratio of the internal flow in the HY-DMA was determined to be 1.6:1. In terms of recovery of the HY-DMA, the highest recovery was obtained at a voltage of 500 V regardless of which type of asbestos was tested. The recovery rate for serpentine was 45.5% and that for amphibole was 34.9%. The coefficients of determination of serpentine (R2 = 0.89) and amphibole (R2 = 0.87) were highly correlated. With respect to the coefficient of variation (CV), the KOFAM demonstrated superior performance over the M7400AD and F-1 methods and showed almost no difference from the PCM method (KOFAM: 22.5%, M7400AD: 32.4%, F-1: 88.8%, and PCM: 21.9%). There was no statistically significant difference between concentration measurements of the KOFAM and PCM analyses. Accordingly, it was concluded that the KOFAM can be used as a superior alternative to conventional fiber measurement methods. The preliminary results support the use of the KOFAM for constant measurement of airborne asbestos concentrations in real time.