Category Archives: RESEARCH ARTICLE

Loss of the adaptor protein Sh3bgrl initiates ovarian fibrosis in zebrafish

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Loss of the adaptor protein Sh3bgrl initiates ovarian fibrosis in zebrafish

Our study shows that knockout of the small adaptor protein SH3-domain-binding glutamate-rich protein-like (Sh3bgrl) causes ovulation and spawning inability in female zebrafish. Mechanistically, sh3bgrl knockout leads to upregulation of genes related to extracellular matrix (ECM) and fiber generation, rendering extra ECM and/or fiber accumulation and deposition in the ovary and the eventual spawning inability.


Ovarian fibrosis is a reproduction obstacle leading to female infertility in vertebrates, but the cause underlying the cellular events is unclear. Here, we found that the small adaptor protein SH3-domain-binding glutamate-rich protein like (Sh3bgrl) plays an important role in female reproduction in zebrafish. Two sh3bgrl mutant alleles that result in sh3bgrl depletion contribute to female spawning inability. Comparative transcriptome analysis revealed that sh3bgrl knockout mechanistically causes the upregulation of genes associated with extracellular matrix (ECM) and fiber generation in the zebrafish ovary. Consequently, extra ECM or fibers accumulate and are deposited in the ovary, resulting in eventual spawning inability. Our findings thus provide insights into understanding the underlying mechanism of infertility by ovarian fibrosis and provide a novel and valuable model to study female reproduction abnormality.

Niche dynamics along two centuries of multiple crayfish invasions

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Niche dynamics along two centuries of multiple crayfish invasions

A historical ecology approach unveiled how the realised niche of a European crayfish species changed since 1850. In response to the introduction of two North American crayfish species, along with a lethal pathogen, its niche shrank and shifted towards more abrupt, headwater environments where the overseas invaders have not arrived.


Abstract

The realised ecological niches of species may change in response to dynamic abiotic and biotic environments, particularly under fast global change. To fully understand the dynamics of niche features and their drivers, it is essential to have a long-term view of species distributions and the factors that may have influenced them. Here, we analysed the distribution and niche dynamics of the Italian crayfish (Austropotamobius fulcisianus) in the Iberian Peninsula over the past 200 years. The Italian crayfish was introduced to Spain in the 16th century, and spread due to multiple stocking events until the 1970s, when two North American crayfish (red swamp crayfish Procambarus clarkii, and signal crayfish Pacifastacus leniusculus) were introduced. Both North American species are carriers of a pathogen (Aphanomyces astaci, the causal agent of crayfish plague) lethal to the Italian crayfish. We hypothesised that the realised niche of the Italian crayfish, both in breadth and in position, has changed over time following changes in its range. The distribution of the Italian crayfish expanded from the mid-19th century until the mid-20th century, in association with an enlargement of its realised niched, mostly towards less abrupt and more coastal-influenced areas. After the introduction of the North American crayfishes, the collapse of the Italian crayfish involved a niche shift towards rough terrains in mountain areas. North American crayfish have eventually occupied most of the Italian crayfish's niche space, with the few no-coexistence areas being relegated to the most abrupt and high-elevation territories. Our historical approach allowed us to document and understand the highly dynamic distribution and niche of the Italian crayfish in the presence of invader counterparts, and to explore the environmental conditions under which their coexistence is minimised.

Batch culture analysis to identify potent organic acids for suppressing ruminal methane production

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Abstract

We performed an in vitro rumen batch culture study to screen 11 commercially available organic acids for methane-suppressing ability and analyzed the rumen microbiota to determine the mode of action of the acids that showed potent methane-suppressing activity. Nine of the 11 acids showed methane-suppressing activity. Maleic anhydride, itaconate, citrate, and fumarate, which showed the highest activity, were further examined. These four acids showed methane-suppressing activity irrespective of the hay-to-concentrate ratios of the substrate. Maleic anhydride and itaconate decreased total gas and short-chain fatty acid production. Maleic anhydride and fumarate increased propionate production, while itaconate increased butyrate production. Maleic anhydride, itaconate, and citrate increased lactate production. Fumarate increased the abundance of bacteria involved in propionate production. Maleic anhydride, itaconate, and citrate increased the abundance of bacteria involved in lactate production. Thus, the results indicate that maleic anhydride, itaconate, and citrate may decrease methane in part by stimulating the acrylate pathway.

Developing models to predict feces and urine excretion in Japanese Black fattening steer by multiple regression analysis

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Abstract

The objective of this study was to develop models for predicting the amount of feces and urine excreted by Japanese Black fattening steer using a dataset of 119 digestion trials for a total of 46 animals. Correlation analysis was used to analyze the relationships between feces and urine excretion and feed intake, feed digestibility, and nitrogen balance. Multiple regression analysis was conducted to develop models for predicting the amount of feces and urine excreted using the explanatory variables selected from various animal and dietary parameters based on P-value (<0.10) and variance inflation factor (<3.0). Resultingly, dry matter intake was a primary predictor of feces excreted. The prediction equation for the amount of feces excretion as a function of body weight, dry matter intake, and calculated total digestible nutrients fits the data well (adjusted coefficient of determination [adj R 2] = 0.519, root mean square error = 1.57). Furthermore, the nitrogen content in the urine was the primary predictor of the urine excretion amount. Thus, the prediction equation for the amount of urine excreted using the nitrogen content in urine yielded a highly accurate model (adj R 2 = 0.813, root mean square error = 4.12).

Characterisation of IL‐1 family members in Sweet syndrome highlights the overexpression of IL‐1β and IL‐1R3 as possible therapeutic targets

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Abstract

Sweet syndrome (SS) as a prototypic neutrophilic dermatosis (NDs) shares certain clinical and histologic features with monogenic auto-inflammatory disorders in which interleukin (IL)-1 cytokine family members play an important role. This has led to the proposal that NDs are polygenic auto-inflammatory diseases and has fuelled research to further understand the role of IL-1 family members in the pathogenesis of NDs. The aim of this study was to characterise the expression of the IL-1 family members IL-1β, IL-36γ, IL-33 and IL-1R3 (IL-1RaP) in SS. The expression profile of IL-1β, IL-33, IL-36γ and their common co-receptor IL-1R3 was analysed by immunohistochemistry, in situ hybridisation and double immunofluorescence (IF) in healthy control skin (HC) and lesional skin samples of SS. Marked overexpression of IL-1β in the dermis of SS (p < 0.001), and a non-significant increase in dermal (p = 0.087) and epidermal (p = 0.345) IL-36γ expression compared to HC was observed. Significantly increased IL-1R3 expression within the dermal infiltrate of SS skin samples (p = 0.02) was also observed, whereas no difference in IL-33 expression was found between SS and HC (p = 0.7139). In situ hybridisation revealed a good correlation between gene expression levels and the above protein expression levels. Double IF identifies neutrophils and macrophages as the predominant sources of IL-1β. This study shows that IL-1β produced by macrophages and neutrophils and IL-1R3 are significantly overexpressed in SS, thereby indicating a potential pathogenic role for this cytokine and receptor in SS.

Single‐cell analysis of human dermal fibroblasts isolated from a single male donor over 35 years

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Single-cell analysis of human dermal fibroblasts isolated from a single male donor over 35 years

The effect of ageing of the same donor on dermal fibroblast heterogeneity was tested in a dermal fibroblast lineage (ASF-4 cell lineage) isolated from the inner side of the upper arm of a healthy male donor (age 36) for 35 years. Single-cell RNA sequencing of cultured cells frozen at Passage 10 at the age of 36 and 72 years showed that genes involved in extracellular matrix remodelling were downregulated with donor ageing, which is a different observation from those reported in the previous studies. Based on the scRNA-Seq results, we identified a cellular senescence-associated gene EFEMP2.


Abstract

The aim of this study is to examine the effects of ageing on dermal fibroblast heterogeneity based on samples obtained from the same donor. We used a dermal fibroblast lineage (named ASF-4 cell lines) isolated from the inner side of the upper arm of a healthy male donor over a 35-year period, beginning at 36 years of age. Because clonal analysis of ASF-4 cell lines demonstrated a donor age-dependent loss of proliferative capacity and acquisition of senescent traits at the single-cell level, cultured cells frozen at passage 10 at ages 36 and 72 years were subjected to single-cell RNA sequencing. Transcriptome analysis revealed an increase in senescent fibroblasts and downregulation of genes associated with extracellular matrix remodelling with ageing. In addition, two putative differentiation pathways, with one endpoint consisting of senescent fibroblasts and the other without, were speculated using a pseudo-time analysis. Knockdown of the characteristic gene of the non-senescent fibroblast cluster endpoint, EFEMP2, accelerated cellular senescence. This was also confirmed in two other normal human dermal fibroblast cell lines. The detection of a common cellular senescence-related gene from single-donor analysis is notable. This study provides new insights into the behaviour of dermal fibroblasts during skin ageing.

A 2‐week time‐restricted feeding attenuates psoriasis‐like lesions with reduced inflammatory cytokines and immunosenescence in mice

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Abstract

Psoriasis, a well-established T-cell mediated dermatosis, exhibits a robust correlation with obesity and systemic inflammation, manifesting psoriasis skin lesions and premature immunosenescence within the peripheral blood and lesion. Intermittent fasting (IF) has exhibited various beneficial effects in reducing inflammation, resisting oxidative stress and slowing ageing, as well as losing weight. A form of IF known as time-restricted feeding (TRF) restricts daily caloric intake within 4–8 h. Nonetheless, the advantageous impacts of TRF on psoriasis still require further verification. We measured the acanthosis in Imiquimod (IMQ)-induced psoriasis mice and evaluated their pathological phenotypes. Our study examined the effects of a 2-week TRF on body weight and metabolic parameters. The subsets of T cells in spleens and skin lesions were accessed by flow cytometry. Cytokines and senescence-associated genes were evaluated by immunofluorescence and RT-qPCR. RNA sequencing was conducted on skin lesions. According to our findings, a 2-week TRF attenuates psoriasis-like lesions in mice with reduced inflammatory cytokines and mitigated immunosenescence. TRF increased the counts of CD4+ Treg cells in skin lesions while reducing the counts of Th2 and Th17 cells in spleens. Furthermore, the administration of TRF resulted in a decrease in the population of CD4+ senescent T cells in both the dermis and spleens, concomitant with the expression of senescence-associated genes in spleen CD4+ T cells. The outcomes mentioned above provide valuable evidence in support of TRF for the management of psoriasis.

Nano‐encapsulated anandamide reduces inflammatory cytokines in vitro and lesion severity in a murine model of cutaneous lupus erythematosus

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Abstract

Cutaneous lupus erythematosus (CLE) is a heterogeneous autoimmune skin disease which occurs independently and in conjunction with systemic lupus erythematosus. Drug development for CLE is severely lacking. Anandamide (AEA) is a primary endocannabinoid which exhibits immunomodulatory effects through mixed cannabinoid receptor agonism. We evaluated AEA as topical treatment for CLE and assessed benefits of nanoparticle encapsulation (AEA-NP) on cutaneous drug penetration, delivery and biological activity. Compared to untreated controls, AEA-NP decreased IL-6 and MCP-1 in UVB-stimulated keratinocytes (p < 0.05) in vitro. In BALB/c mice, AEA-NP displayed improved cutaneous penetration, extended release and persistence of AEA in the follicular unit extending to the base after 24 h. Utilizing the MRL-lpr lupus murine model, twice weekly treatment of lesions with topical AEA-NP for 10 weeks led to decreased clinical and histologic lesion scores compared to unencapsulated AEA and untreated controls (p < 0.05). Prophylactic application of AEA-NP to commonly involved areas on MRL-lpr mice similarly resulted in decreased clinical and histologic scores when compared to controls (p < 0.05), and reduced C3 and IBA-1 in lesional tissue (p < 0.05). The demonstrated clinical and immunomodulatory effects of treatment with AEA support its potential as therapy for CLE. This work also suggests that encapsulation of AEA improves penetration and treatment efficacy. Future studies will be conducted to assess full therapeutic potential.

Altered actin dynamics is possibly implicated in the inhibition of mechanical stimulation‐induced dermal fibroblast differentiation into myofibroblasts

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Abstract

The formation of hypertrophic scars and keloids is strongly associated with mechanical stimulation, and myofibroblasts are known to play a major role in abnormal scar formation. Wounds in patients with neurofibromatosis type 1 (NF1) become inconspicuous and lack the tendency to form abnormal scars. We hypothesized that there would be a unique response to mechanical stimulation and subsequent scar formation in NF1. To test this hypothesis, we investigated the molecular mechanisms of differentiation into myofibroblasts in NF1-derived fibroblasts and neurofibromin-depleted fibroblasts and examined actin dynamics, which is involved in fibroblast differentiation, with a focus on the pathway linking LIMK2/cofilin to actin dynamics. In normal fibroblasts, expression of α-smooth muscle actin (α-SMA), a marker of myofibroblasts, significantly increased after mechanical stimulation, whereas in NF1-derived and neurofibromin-depleted fibroblasts, α-SMA expression did not change. Phosphorylation of cofilin and subsequent actin polymerization did not increase in NF1-derived and neurofibromin-depleted fibroblasts after mechanical stimulation. Finally, in normal fibroblasts treated with Jasplakinolide, an actin stabilizer, α-SMA expression did not change after mechanical stimulation. Therefore, when neurofibromin was dysfunctional or depleted, subsequent actin polymerization did not occur in response to mechanical stimulation, which may have led to the unchanged expression of α-SMA. We believe this molecular pathway can be a potential therapeutic target for the treatment of abnormal scars.

Compared to antibiotic, feeding peony byproducts can improve growth performance, rumen fermentation, slaughter parameters, and meat quality of lambs

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Abstract

Our objective was to determine effects of feeding lamb's peony byproducts, including stem and leaves (PSL), root (PR), and seeds meal (PSM), on growth, rumen fermentation, slaughter parameters, and meat quality. Sixty-four lambs (14.0 ± 2.1 kg) were allocated into eight treatments based on BW: no additives (CON), 0.15% aureomycin (CONA), low/high levels of PSL (5%/10% PSL replaced 5%/10% Chinese hay), PR (basal diet with 0.5%/1.0% PR), PSM (5%/10% PSM replaced 5%/10% soybean meal). Growth, digestibility, and rumen fermentation had dose responses whereas slaughter parameters, meat quality, or amino acids indexes were not. Peony byproducts increased DMI (p < 0.001) compared to CON, but higher levels were more advantageous (p = 0.003). However, low levels of peony byproducts decreased the NH3-N concentration, but increased total volatile fatty acids mole percent more than high levels of that (p < 0.001). All peony byproducts increased dressing percentage (p < 0.05), increased pH and tenderness than CON (p < 0.05). In addition, PSL and PSM improved amino acid profiles of meat compared to CON, and were even better than CONA (p < 0.05). Therefore, peony byproducts not only improved animal growth but also reduced the frequency of antibiotic use in animal feeding.