Dual-color live imaging unveils stepwise organization of multiple basal body arrays by cytoskeletons.

For mucociliary clearance of pathogens, tracheal multiciliated epithelial cells (MCCs) organize coordinated beating of cilia, which originate from basal bodies (BBs) with basal feet (BFs) on one side. To clarify the self-organizing mechanism of coordinated intracellular BB-arrays composed of a well-ordered BB-alignment and unidirectional BB-orientation, determined by the direction of BB to BF, we generated double transgenic mice with GFP-centrin2-labeled BBs and mRuby3-Cep128-labeled BFs for long-term, high-resolution, dual-color live-cell imaging in primary-cultured tracheal MCCs. At early timepoints of MCC differentiation, BB-orientation and BB-local alignment antecedently coordinated in an apical microtubule-dependent manner. Later during MCC differentiation, fluctuations in BB-orientation were restricted, and locally aligned BB-arrays were further coordinated to align across the entire cell (BB-global alignment), mainly in an apical intermediate-sized filament-lattice-dependent manner. Thus, the high coordination of the BB-array was established for efficient mucociliary clearance as the primary defense against pathogen infection, identifying apical cytoskeletons as potential therapeutic targets.

In Vitro Antioxidant and Antiaging Activities of Collagen and Its Hydrolysate from Mackerel Scad Skin (Decapterus macarellus).

The skin of mackerel scad fish (Decapterus macarellus) is a new source for pepsin-soluble collagen and its hydrolysate, both of which have never been explored. This study aims to characterize and determine the in vitro antioxidant, antiglycation, and antityrosinase activity of pepsin-soluble collagen (PSC) and hydrolyzed collagen (HC) from mackerel scad skin. PSC was extracted using 0.5 M acetic acid containing 0.1% pepsin for 48 h at 4 °C. The obtained PSC was then hydrolyzed with collagenase type II (6250 U/g) to produce HC. The PSC yield obtained was 6.39 ± 0.97%, with a pH of 6.76 ± 0.18, while the HC yield was 96% from PSC. SDS-PAGE and Fourier Transform Infrared (FTIR) analysis showed the typical features of type I collagen. HC demonstrated high solubility (66.75-100%) throughout the entire pH range (1-10). The PSC and HC from mackerel scad skin showed antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH), with IC50 values of 148.55 ± 3.14 ppm and 34.966 ± 0.518 ppm, respectively. In the antiglycation test, PSC had an IC50 value of 239.29 ± 15.67 ppm, while HC had an IC50 of 68.43 ± 0.44 ppm. PSC also exhibited antityrosinase activity, with IC50 values of 234.66 ± 0.185 ppm (on the L-DOPA substrate), while HC had an IC50 value of 79.35 ± 0.5 ppm. Taken together, these results suggest that the skin of mackerel scad fish has potential antiaging properties and can be further developed for pharmaceutical and cosmetic purposes.

Avenolide, a Streptomyces hormone controlling antibiotic production in Streptomyces avermitilis.

Gram-positive bacteria of the genus Streptomyces are industrially important microorganisms, producing >70% of commercially important antibiotics. The production of these compounds is often regulated by low-molecular-weight bacterial hormones called autoregulators. Although 60% of Streptomyces strains may use γ-butyrolactone-type molecules as autoregulators and some use furan-type molecules, little is known about the signaling molecules used to regulate antibiotic production in many other members of this genus. Here, we purified a signaling molecule (avenolide) from Streptomyces avermitilis--the producer of the important anthelmintic agent avermectin with annual world sales of $850 million--and determined its structure, including stereochemistry, by spectroscopic analysis and chemical synthesis as (4S,10R)-10-hydroxy-10-methyl-9-oxo-dodec-2-en-1,4-olide, a class of Streptomyces autoregulator. Avenolide is essential for eliciting avermectin production and is effective at nanomolar concentrations with a minimum effective concentration of 4 nM. The aco gene of S. avermitilis, which encodes an acyl-CoA oxidase, is required for avenolide biosynthesis, and homologs are also present in Streptomyces fradiae, Streptomyces ghanaensis, and Streptomyces griseoauranticus, suggesting that butenolide-type autoregulators may represent a widespread and another class of Streptomyces autoregulator involved in regulating antibiotic production.

Oral intake of collagen hydrolysate from mackerel scad (Decapterus macarellus) attenuates skin photoaging by suppressing the UVB-induced expression of MMP-1 and IL-6.

OBJECTIVES: Excessive skin exposure to UVB radiation can induce photoaging caused by an imbalance in oxidative stress and inflammatory responses, damaging the skin's structure and surface layer. A previous study revealed that collagen hydrolisate extracted from the skin of mackarel scads (Decapterus macarellus) had antiaging properties that were tested in vitro, which serves as a foundation for a subsequent study of its use in vivo. This study aimed at investigating the repair effect of the mackerel scad's skin collagen hydrolysate (MSS-CH) in photoaging conditions in a mouse model. METHODS: MSS-CH was given orally in mice model of skin photoaging under chronic exposure to UVB irradiation for 12 weeks. Morphological and histological changes on the skin were evaluated using SEM and HE staining, along with the measurement of the expression of matrix metalloproteinases (MMP-1) and cytokine pro-inflammatory interleukin 6 (IL-6) using ELISA. RESULTS: MSS-CH inhibits the occurrence of epidermal thickening and damage to the dermal layer of the skin. As a result, it restores the epidermis' barrier function and reduces surface damage caused by photoaging. The skin of the MSS-CH treated group exhibited improved physical appearance with reduced fine lines, wrinkles, and enhanced smoothness. Additionally, administering MSS-CH to the mice groups reduced the expression of MMP-1 and IL-6 in UVB-exposed skin. CONCLUSIONS: Altogether, this in vivo study demonstrates the photoaging-protective properties of CH-MSS, aligning with previous in vitro data. Thus, MSS-CH emerges as a strong candidate for use as an ingredient in nutraceuticals and biocosmetics.

Metabolomics-driven strain improvement: A mini review.

In recent years, mass spectrometry-based metabolomics has been established as a powerful and versatile technique for studying cellular metabolism by comprehensive analysis of metabolites in the cell. Although there are many scientific reports on the use of metabolomics for the elucidation of mechanism and physiological changes occurring in the cell, there are surprisingly very few reports on its use for the identification of rate-limiting steps in a synthetic biological system that can lead to the actual improvement of the host organism. In this mini review, we discuss different strategies for improving strain performance using metabolomics data and compare the application of metabolomics-driven strain improvement techniques in different host microorganisms. Finally, we highlight several success stories on the use of metabolomics-driven strain improvement strategies, which led to significant bioproductivity improvements.

Planar cell polarity induces local microtubule bundling for coordinated ciliary beating.

Multiciliated cells (MCCs) in tracheas generate mucociliary clearance through coordinated ciliary beating. Apical microtubules (MTs) play a crucial role in this process by organizing the planar cell polarity (PCP)-dependent orientation of ciliary basal bodies (BBs), for which the underlying molecular basis remains elusive. Herein, we found that the deficiency of Daple, a dishevelled-associating protein, in tracheal MCCs impaired the planar polarized apical MTs without affecting the core PCP proteins, causing significant defects in the BB orientation at the cell level but not the tissue level. Using live-cell imaging and ultra-high voltage electron microscope tomography, we found that the apical MTs accumulated and were stabilized by side-by-side association with one side of the apical junctional complex, to which Daple was localized. In vitro binding and single-molecule imaging revealed that Daple directly bound to, bundled, and stabilized MTs through its dimerization. These features convey a PCP-related molecular basis for the polarization of apical MTs, which coordinate ciliary beating in tracheal MCCs.

Daple deficiency causes hearing loss in adult mice by inducing defects in cochlear stereocilia and apical microtubules.

The V-shaped arrangement of hair bundles on cochlear hair cells is critical for auditory sensing. However, regulation of hair bundle arrangements has not been fully understood. Recently, defects in hair bundle arrangement were reported in postnatal Dishevelled-associating protein (ccdc88c, alias Daple)-deficient mice. In the present study, we found that adult Daple-/- mice exhibited hearing disturbances over a broad frequency range through auditory brainstem response testing. Consistently, distorted patterns of hair bundles were detected in almost all regions, more typically in the basal region of the cochlear duct. In adult Daple-/- mice, apical microtubules were irregularly aggregated, and the number of microtubules attached to plasma membranes was decreased. Similar phenotypes were manifested upon nocodazole treatment in a wild type cochlea culture without affecting the microtubule structure of the kinocilium. These results indicate critical role of Daple in hair bundle arrangement through the orchestration of apical microtubule distribution, and thereby in hearing, especially at high frequencies.

Multiciliated cell basal bodies align in stereotypical patterns coordinated by the apical cytoskeleton.

Multiciliated cells (MCCs) promote fluid flow through coordinated ciliary beating, which requires properly organized basal bodies (BBs). Airway MCCs have large numbers of BBs, which are uniformly oriented and, as we show here, align linearly. The mechanism for BB alignment is unexplored. To study this mechanism, we developed a long-term and high-resolution live-imaging system and used it to observe green fluorescent protein-centrin2-labeled BBs in cultured mouse tracheal MCCs. During MCC differentiation, the BB array adopted four stereotypical patterns, from a clustering "floret" pattern to the linear "alignment." This alignment process was correlated with BB orientations, revealed by double immunostaining for BBs and their asymmetrically associated basal feet (BF). The BB alignment was disrupted by disturbing apical microtubules with nocodazole and by a BF-depleting Odf2 mutation. We constructed a theoretical model, which indicated that the apical cytoskeleton, acting like a viscoelastic fluid, provides a self-organizing mechanism in tracheal MCCs to align BBs linearly for mucociliary transport.