Dorsal Tissue Research Articles

Gene profiling in dorso-ventral patterning of mouse tongue development.

The tongue is a muscular fleshy organ in the oral cavity that is anatomically divided into the dorsal, ventral, anterior, and posterior part. The intricate tissue organisation and diverse origins of the tongue make it a complex organ of the oral cavity. To reveal the signalling molecules involved in the formation of the dorsal and ventral parts of the tongue through microarray analysis. Dorsal and ventral tongue tissues were isolated from embryonic day 14 mice by micro-dissection. RNA was extracted from the dorsal and ventral tongue tissues separately for microarray analysis. Microarray data were confirmed by quantitative reverse transcription polymerase chain reaction and whole-mount in situ hybridisation. Microarray analysis revealed expression of 33,793 genes. Of these, 931 genes were found to be equally expressed in both the dorsal and ventral parts of the tongue. On limiting the fold-change cut-off to over 1.5-fold, 725 genes were expressed over 1.5-fold in the ventral part and 1,672 in the dorsal part of the tongue. The qPCR and whole-mount in situ hybridisation revealed the expressions of angiopoietin 2 (Angpt2), fibroblast growth factor 18 (Fgf18), mesenchyme homeobox gene1 (Meox1), and SPARC-related modular calcium binding 2 (Smoc2) in the ventral part of the tongue. Numerous signalling molecules can be selected from our microarray results to examine their roles in tongue development and disease model systems. In the near future, the selection of candidate genes and their functional evaluations will be performed through loss- and gain-of-function mutation studies.

Biomechanical Comparison of a Headless Compression Screw Fastener and AO Cortical Bone Screw for Fixation of a Simulated Equine Third Carpal Bone Slab Fracture

Frontal plane slab fractures account for the majority of third carpal bone (C3) fractures in racing and performance horses. Recommended treatment is stabilization with a lagged AO cortical screw. Associated complications are fragment splitting, fragment spinning, and irritation of dorsal soft tissue structures. A novel, headless, cannulated screw with interlocking threads the Headless Compression Screw Fastener (HCSF) has been developed to resist multidirectional forces and bending moments; however, it has not been applied in the horse. Simulated C3 frontal plane slab fractures were created in nine paired carpi from equine cadaver limbs, fixed with either the HCSF or AO cortical bone screw, and loaded in shear to failure. The effect of screw type on stiffness, maximum load to failure, and yield load was assessed in separate linear mixed models. No significant (P< .05) difference between screw types was detected in terms of maximum load to failure (P= .084), stiffness (P= .26), or yield load (P= .088). Mode of failure was screw bending in all specimens. For some samples in both groups, failure was associated with the sagittal fracture at the screw-bone interface. The HCSF was successfully used to repair simulated third carpal bone fractures. The different head and thread pitches of the HCSF effectively compressed the fracture. The headless design eliminates the need for counter sinking. There was no significant difference in maximum load to failure, stiffness, nor yield load compared to the cortical screws. These results invite clinical application to be investigated.

NF-κB and AP-1 are required for the lipopolysaccharide-induced expression of MCP-1, CXCL1, and Cx43 in cultured rat dorsal spinal cord astrocytes.

TLR4 and Cx43 signaling in dorsal spinal cord has been shown to be involved in the development of neuropathic pain. However, it is not clear whether TLR4 signaling is associated with the expression of MCP-1, CXCL1, and Cx43 in LPS (lipopolysaccharide)-treated rat dorsal spinal cord astrocytes under in vitro condition. In the present study, we found that TLR4 antagonist TAK-242 significantly inhibited LPS-induced MCP-1, CXCL1, and Cx43 expression, suggesting the role of TLR4 in response to LPS in cultured dorsal spinal cord astrocytes. Application of TAK-242 significantly blocked LPS-induced NF-κB and AP-1 activity and the expression of MCP-1, CXCL1 and Cx43. Furthermore, NF-κB inhibitor PDTC and AP-1 inhibitor SR11302 significantly blocked LPS-induced MCP-1, CXCL1, and Cx43 expression. DNA-binding activity of NF-κB, its effect on MCP-1 expression was suppressed by PDTC and SR11302. On the other hand, DNA-binding activity of AP-1, its effect on CXCL1 or Cx43 expression was also suppressed by PDTC and SR11302. In addition, PDTC was found to inhibit the nuclear translocation of AP-1 and the expression of c-Jun induced by LPS, which suggested that NF-κBp65 is essential for the AP-1 activity. Similarly, SR11302 significantly blocked LPS-induced the nuclear translocation of NF-κBp65 and the expression of NF-κBp65 induced by LPS. Pretreatment with CBX, Gap26, or Gap19 (Cx43 blockers) significantly inhibited abnormal astrocytic hemichannel opening and chemokines (MCP-1 and CXCL1) release in LPS-stimulated astrocytes. In summary, cell culture experiments revealed that LPS stimulation could evoke TLR4 signaling with the subsequent activation of NF-κB and AP-1, resulting in the expression of MCP-1, CXCL1, and Cx43. TLR4 activation increased Cx43 hemichannel, but not gap-junction activities and induced the release of the MCP-1 and CXCL1 from astrocytes via Cx43 hemichannel. These findings may help us to understand the role of astrocytic signaling in inflammatory response within dorsal spinal cord tissue.

138 Single-cell RNA-seq of dorsal root ganglion reveals neuroinflammatory process in atopic dermatitis

Atopic dermatitis is a chronic inflammatory skin disease, the pathology of which is characterized by barrier dysfunction, microbiome shift, immune dysregulation initiated by a severe itching sensation. Because of its pathophysiological and clinical significance, research on understanding the mechanism of itch and finding new therapeutic targets on it has been highlighted recently. To dissect the cellular/molecular changes associated with itch, we obtained a total of six mouse dorsal root ganglion tissues; three from atopic dermatitis-induced NC/Nga mice and another three from control mice, and performed single-cell RNA sequencing. Among non-neuronal cells, proportions of neutrophil and macrophage increased in atopic dermatitis, which implies the presence of neuroinflammation. Non-peptidergic neurons, which have nociceptors transmitting itching sensation, occupy a larger proportion among neurons in atopic dermatitis, compared to the control. As the first single-cell transcriptome analysis of atopic dermatitis mouse DRG, our study provides insight into how itching sensation manifests itself on a single cell level in atopic dermatitis.

Analyzing the Influence of Imaging Resolution on Polarization Properties of Scattering Media Obtained From Mueller Matrix

The Mueller matrix contains abundant micro- and even nanostructural information of media. Especially, it can be used as a powerful tool to characterize anisotropic structures quantitatively, such as the particle size, density, and orientation information of fibers in the sample. Compared with unpolarized microscopic imaging techniques, Mueller matrix microscopy can also obtain some essential structural information about the sample from the derived parameters images at low resolution. Here, to analyze the comprehensive effects of imaging resolution on polarization properties obtained from the Mueller matrix, we, first, measure the microscopic Mueller matrices of unstained rat dorsal skin tissue slices rich in collagen fibers using a series of magnifications or numerical aperture (NA) values of objectives. Then, the first-order moments and image texture parameters are quantified and analyzed in conjunction with the polarization parameter images. The results show that the Mueller matrix polar decomposition parameters diattenuation D, linear retardance δ, and depolarization Δ images obtained using low NA objective retain most of the structural information of the sample and can provide fast imaging speed. In addition, the scattering phase function analysis and Monte Carlo simulation based on the cylindrical scatterers reveal that the diattenuation parameter D images with different imaging resolutions are expected to be used to distinguish among the fibrous scatterers in the medium with different particle sizes. This study provides a criterion to decide which structural information can be accurately and rapidly obtained using a transmission Mueller matrix microscope with low NA objectives to assist pathological diagnosis and other applications.

Intrathecal liproxstatin-1 delivery inhibits ferroptosis and attenuates mechanical and thermal hypersensitivities in rats with complete Freund's adjuvant-induced inflammatory pain

Previous studies have confirmed the relationship between iron-dependent ferroptosis and a peripheral nerve injury-induced neuropathic pain model. However, the role of ferroptosis in inflammatory pain remains inconclusive. Therefore, we aimed to explore whether ferroptosis in the spinal cord and dorsal root ganglion contributes to complete Freund's adjuvant (CFA)-induced painful behaviors in rats. Our results revealed that various biochemical and morphological changes were associated with ferroptosis in the spinal cord and dorsal root ganglion tissues of CFA rats. These changes included iron overload, enhanced lipid peroxidation, disorders of anti-acyl-coenzyme A synthetase long-chain family member 4 and glutathione peroxidase 4 levels, and abnormal morphological changes in mitochondria. Intrathecal treatment of liproxstatin-1 (a ferroptosis inhibitor) reversed these ferroptosis-related changes and alleviated mechanical and thermal hypersensitivities in CFA rats. Our study demonstrated the occurrence of ferroptosis in the spinal cord and dorsal root ganglion tissues in a rodent model of inflammatory pain and indicated that intrathecal administration of ferroptosis inhibitors, such as liproxstatin-1, is a potential therapeutic strategy for treating inflammatory pain.

Long Non-Coding RNA and mRNA Profiles in the Spinal Cord of Rats with Resiniferatoxin-Induced Neuropathic Pain.

PurposeThe ultrapotent transient receptor potential vanilloid 1 (TRPV1) agonist resiniferatoxin (RTX) induces small-fiber sensory neuropathy, which has been widely used model of postherpetic neuralgia to study mechanisms of neuropathic pain and new analgesics. The long non-coding RNA (lncRNA) and mRNA expression profiles in spinal dorsal horn tissues of rats six weeks after RTX injection to identify new RNAs related to neuropathic pain.MethodsMicroarray technology was applied to determine lncRNA expressions in spinal dorsal horn samples of adult rats 6 weeks after treatment with RTX or vehicle. The lncNA/mRNA co-expression network was constructed, and differential expression patterns of lncRNA and mRNA in RTX-treated rats were identified. Differential expressions of lncRNAs and mRNAs between RTX-treated samples and control samples were examined by RT-qPCR.ResultsMicroarray analyses showed that 745 mRNA and 139 lncRNAs were upregulated, whereas 590 mRNA and 140 lncRNAs were downregulated in spinal dorsal horn tissues after RTX exposure. TargetScan was used to predict mRNA targets for these lncRNAs, which showed that the transcripts with multiple predicted target sites were related to neurologically important pathways. In addition, differential expressions of lncRNA (ENSRNOG00000022535, ENSRNOG00000042027, NR_027478, NR_030675) and Apobec3b mRNA in spinal cord tissue samples were validated, which confirmed the microarray data. The association between NR_030675 and Apobec3b levels was confirmed, which may be related to neuropathic pain.ConclusionOur study reveals lncRNA and mRNA of molecule targets that are enriched in the spinal cord dorsal horn and provides new information for further investigation on the mechanisms and therapeutics of neuropathic pain.

Trophic Structure and Diet of Predatory Teleost Fishes in a Tropical Demersal Shelf Ecosystem

Predatory fishes are a major component of many tropical fisheries, although little is known about their diet and trophic structure, which can hinder effective management. We used stable isotopes δ15N and δ13C in conjunction with dietary prey items of five fishes (Lutjanus sebae, Lethrinus punctulatus, Epinephelus areolatus, Epinephelus multinotatus, and Plectropomus maculatus) to describe the diet and trophic structure across this assemblage. A total of 153 isotope and 87 stomach content samples were collected at two locations that were ≈30 km apart, over two sampling trips, separated by three months. There was clear separation of species’ mean δ15N and δ13C values in isotopic space; the highest mean δ15N was exhibited by E. multinotatus (13.50 ± 0.11 SE) and the lowest was L. punctulatus (11.05 ± 0.13). These two species had the lowest overlap of isotopic niche space, whereas the highest overlap occurred between L. sebae and P. maculatus. δ15N increased with fish body size for all species. However, body size was not significantly related to δ13C values for any species. There was a notable shift in both δ15N and δ13C between sampling trips, with δ13C being more depleted in the second trip. There was also a difference in δ13C between locations for all species, suggesting localised foraging. A multiple tissue comparison for L. sebae indicated positive relationships for both δ15N and δ13C between dorsal fin and muscle tissue. Mean δ15N values were the same for both fin (12.1 ± 0.10 SE) and muscle tissue (12.1 ± 0.09 SE), although δ13C was more enriched in fin (-15.6 ± 0.14 SE) compared to muscle tissue (-17.3 ± 0.11 SE). The most common dietary items across species were teleosts and crustaceans, which was consistent with isotope data indicating a reliance on a demersal food web (δ13C values ranging from -18 to -15‰). The results from our study indicate a dynamic spatio-temporal trophic structure and diet for commercially important demersal species and highlight the benefits of a multi-facetted sampling approach.

Transcriptomics-Based Study of Differentially Expressed Genes Related to Fat Deposition in Tibetan and Yorkshire Pigs.

Fat deposition traits are one of the key factors in pig production and breeding. The fat deposition capacity of pigs mainly affects the quality of pork and pig productivity. The aim of this study was to analyze the differential expression of mRNA levels in dorsal adipose tissue of Tibetan and York pigs at different growth stages using transcriptomic data to estimate key genes that regulate fat deposition in pigs. The results showed that a total of 32,747 positively expressed genes were present in the dorsal adipose tissue of the two breeds. Differentially expressed gene (DEG) screening of multiple combinations between the two breeds yielded 324 DEGS. Gene ontology (GO) biofunctional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that these DEGS were mainly involved in lipid metabolic pathways, steroid biosynthetic pathways and lipid biosynthetic processes, sterol biosynthetic processes, brown adipocyte differentiation, and other pathways related to lipid deposition and metabolism. The results showed that ACACA, SLC2A4 and THRSP genes positively regulated the lipid deposition ability and CHPT1 gene negatively regulated the lipid deposition ability in pigs. The results of this experiment suggest a theoretical basis for further studies on the regulatory mechanisms of fat deposition in pigs.

Inhibitory effect of Isatis tinctoria L. water extract on DNCB-induced atopic dermatitis in BALB/c mice and HaCaT cells.

BackgroundIsatis tinctoria L (PLG) is a medicinal herb from the roots of Isatis indigotica Fort (Family Cruciferae). Previous studies have shown that PLG has anti-inflammatory and therapeutic effects against conditions such as acute and chronic hepatitis, various respiratory inflammations, and cancer. The purpose of this study was to define the pharmacological effects of PLG on inflammatory reactions and skin hyperkeratosis, which are the main symptoms of atopic dermatitis (AD), in vivo and in vitro.MethodsFor the AD in vivo experiment, 2,4-dinitrochlorobenzene (DNCB) induction and oral administration of PLG were performed on male BALB/c mice for four weeks. For in vitro experiments, keratinocytes were activated using TNF-α/IFN-γ in cultured human keratinocyte (HaCaT) cells. PLG inhibited inflammatory chemokine production and blocked the nuclear translocation of NF-κB p65 in activated keratinocytes.ResultsAs a result of oral administration of PLG, dermis and epidermis thickening, as well as eosinophil and mast cell infiltration, were attenuated in AD skin lesions. In addition, the levels of immunoglobulin E (IgE), pro-inflammatory cytokines, and the MAPK/NF-κB signaling pathway were decreased in serum and dorsal skin tissues. Furthermore, PLG inhibited inflammatory chemokine production and blocked the nuclear translocation of NF-κB p65 in activated keratinocytes. In addition, epigoitrin and adenosine, the standard compounds of PLG, were identified as candidate AD compounds.ConclusionsThese results indicate that PLG is a potent therapeutic agent for attenuating symptoms of AD.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13020-022-00624-5.

Influence of Touch-Spun Nanofiber Diameter on Contact Guidance during Peripheral Nerve Repair.

Peripheral nerve regeneration across large gaps remains clinically challenging and scaffold design plays a key role in nerve tissue engineering. One strategy to encourage regeneration has utilized nanofibers or conduits to exploit contact guidance within the neural regenerative milieu. Herein, we report the effect of nanofiber topography on two key aspects of regeneration: Schwann cell migration and neurite extension. Substrates possessing distinct diameter distributions (300 ± 40 to 900 ± 70 nm) of highly aligned poly(ε-caprolactone) nanofibers were fabricated by touch-spinning. Cell migratory behavior and contact guidance were then evaluated both at the tissue level using dorsal root ganglion tissue explants and the cellular level using dissociated Schwann cells. Explant studies showed that Schwann cells emigrated significantly farther on fibers than control. However, both Schwann cells and neurites emigrated from the tissue explants directionally along the fibers regardless of their diameter, and the data were characterized by high variation. At the cellular level, dissociated Schwann cells demonstrated biased migration in the direction of fiber alignment and exhibited a significantly higher biased velocity (0.2790 ± 0.0959 μm·min-1) on 900 ± 70 nm fibers compared to other nanofiber groups and similar to the velocity found during explant emigration on 900 nm fibers. Therefore, aligned, nanofibrous scaffolds of larger diameters (900 ± 70 nm) may be promising materials to enhance various aspects of nerve regeneration via contact guidance alone. While cells track along with the fibers, this contact guidance is bidirectional along the fiber, moving in the plane of alignment. Therefore, the next critical step to direct regeneration is to uncover haptotactic cues that enhance directed migration.

Inhibitory Effects of Donkey Hide Gelatin on DNCB-Induced Atopic Dermatitis in NC/Nga Mice.

The increase of atopic dermatitis has led to higher socio-economic cost and raised a need for alternative medicine as novel therapeutic agents. In this study, we aimed to evaluate the inhibitory effects of Donkey Hide Gelatin (DHG) water extract on DNCB-induced atopic dermatitis in NC/Nga mice and on tumor necrosis factor (TNF)-α/interferon (IFN)-γ-treated keratinocytes and to investigate its underlying molecular mechanisms. NC/Nga mice were induced by DNCB, administered Dexamethasone (3 mg/kg) or DHG water extracts (100–400 mg/kg) for 3 weeks. The skin symptom score, serum IgE and immune cells were measured, the ALN, spleen and dorsal skin tissue were extracted for FACS, quantitative real-time PCR and histology analysis. In vitro, HaCaT cells were induced by TNF-α/IFN-γ, the levels of pro-inflammatory cytokines and chemokines and its underlying mechanism were measured by ELISA and Western blot. As a result, DHG groups showed a significant decrease in the skin symptom score and the immune cell absolute number. It also showed a marked reduction of allergic and the levels of neutrophils and eosinophils in histology analysis. In TNF-α/IFN-γ induced HaCaT cells, DHG showed inhibition effects on IL-6, IL-8, TARC and RANTES, it also downregulated the expression of ICAM-1 and COX-2, up-regulated the expression of Filaggrin. Furthermore, DHG suppressed the activation of NF-κB and mitogen-activated protein kinases (MAPK) signaling pathway induced by TNF-α/IFN-γ. Taken together, DHG maybe a potential therapeutic agent or supplement for skin inflammatory disease such as atopic dermatitis.

In Ovo Injection of CHIR-99021 Promotes Feather Follicle Development via Modulating the Wnt Signaling Pathway and Transcriptome in Goose Embryos (Anser cygnoides).

Feather performs important physiological functions in birds, and it is also one of the economic productions in goose farming. Understanding and modulating feather follicle development during embryogenesis are essential for bird biology and the poultry industry. CHIR-99021 is a potent Wnt/β-catenin signaling pathway activator associated with feather follicle development. In this study, goose embryos (Anser cygnoides) received an in ovo injection of CHIR-9902, which was conducted at the beginning of feather follicle development (E9). The results showed that feather growth and feather follicle development were promoted. The Wnt signaling pathway was activated by the inhibition of GSK-3β. Transcriptomic analyses showed that the transcription changes were related to translation, metabolism, energy transport, and stress in dorsal tissue of embryos that received CHIR-99021, which might be to adapt and coordinate the promoting effects of CHIR-99021 on feather follicle development. This study suggests that in ovo injection of CHIR-99021 is a potential strategy to improve feather follicle development and feather-related traits for goose farming and provides profiling of the Wnt signaling pathway and transcriptome in dorsal tissue of goose embryos for further understanding of feather follicle development.

A Novel Oral Endoscopic Biopsy Procedure to Obtain Rumen Epithelial Samples.

Most in vivo studies related to ruminal development in calves use invasive techniques involving rumen-fistulated or euthanized animals. In consideration of animal welfare, we developed an oral endoscopic biopsy procedure to allow the obtaining of rumen epithelial samples, thus serving as an alternative for measuring the height and width of rumen papillae in calves in a safe, quick, and efficient manner that allows the slaughtering of calves to be avoided. This procedure was tested on 12 Brangus crossbred calves randomly distributed in two groups, with one fed a meal starter and the other an extruded starter feed. Calves underwent a 12-h fasting period, were restrained in a squeeze chute, administered a dose of atropine, and sedated with xylazine before the oral endoscopic biopsy procedure. A 120 cm long Olympus® oral flexible video endoscope and forceps were used to collect cranial–dorsal sac rumen epithelial tissue samples of approximately 0.5 mm. Endoscopy was successful in all 12 calves and the collected tissue samples were placed in formalin (10%) for further processing for obtaining rumen papillae measurements. Consumption of the extruded starter feed resulted in the increased (p = 0.035) width of rumen papillae. The oral endoscopic biopsy procedure implemented in this study was demonstrated to be successful and is thus an alternative technique for studying rumen epithelial development and morphometric alterations in calf rumen tissue.

MDL-800, the SIRT6 Activator, Suppresses Inflammation via the NF-κB Pathway and Promotes Angiogenesis to Accelerate Cutaneous Wound Healing in Mice.

Sirtuin 6 (SIRT6) is an NAD+-dependent deacetylase belonging to the sirtuin family. It has been shown to participate in wound healing and some inflammation-related disorders. However, the effect of MDL-800, a highly efficient and selective SIRT6 activator, on wound healing and inflammation has not been reported. Therefore, this study investigated whether MDL-800 confers anti-inflammatory effects and promotes wound healing and uncovered the molecular mechanisms involved. This was achieved using mouse models of full-thickness wounds. Results showed that MDL-800 significantly downregulated inflammation by attenuating the release of inflammatory mediators and improved collagen deposition and neovascularization of wounds, thereby accelerating cutaneous wound healing. Furthermore, MDL-800 significantly downregulated expression levels of TNF-α and IL-6 in the dorsal skin tissue of mice via the NF-κB pathway. These results demonstrated that MDL-800 exerted anti-inflammatory and prohealing effects, indicating that the SIRT6/NF-κB/IκB signaling pathway may play an important role in wound healing.

A human coronavirus OC43-derived polypeptide causes neuropathic pain.

Human coronaviruses have been recently implicated in neurological sequelae by insufficiently understood mechanisms. We here identify an amino acid sequence within the HCoV‐OC43 p65‐like protein homologous to the evolutionarily conserved motif of myelin basic protein (MBP). Because MBP‐derived peptide exposure in the sciatic nerve produces pronociceptive activity in female rodents, we examined whether a synthetic peptide derived from the homologous region of HCoV‐OC43 (OC43p) acts by molecular mimicry to promote neuropathic pain. OC43p, but not scrambled peptides, induces mechanical hypersensitivity in rats following intrasciatic injections. Transcriptome analyses of the corresponding spinal cords reveal upregulation of genes and signaling pathways with known nociception‐, immune‐, and cellular energy‐related activities. Affinity capture shows the association of OC43p with an Na+/K+‐transporting ATPase, providing a potential direct target and mechanistic insight into virus‐induced effects on energy homeostasis and the sensory neuraxis. We propose that HCoV‐OC43 polypeptides released during infection dysregulate normal nervous system functions through molecular mimicry of MBP, leading to mechanical hypersensitivity. Our findings might provide a new paradigm for virus‐induced neuropathic pain.

Preservation Rhinoplasty: Open or Closed?

Preservation rhinoplasty (PR) is an evolving philosophy. The open approach was initially utilized, but the author felt a closed approach might be of benefit in certain patients. A total 162 primary rhinoplasty cases were studied retrospectively between May and November 2020. One hundred cases had at least 1 year of follow-up. Patients had follow-up at 1 week, 1 month, 3 months, and 1 year after surgery. Technical details were recorded, including dissection planes, preservation of the dorsum (DP) vs component reductions, surface vs foundational DP techniques, and open vs closed approach. One hundred patients had at least 1 year of follow-up. Fifty-six patients underwent an open approach and 44 a closed approach. Eighty-three patients had preservation of the dorsal soft tissue envelope. All patients who underwent a closed approach had preservation of the dorsal soft tissue envelope. Sixty-seven patients underwent DP, with 38 receiving surface techniques and 29 undergoing impaction techniques. Thirty-three patients underwent structural rhinoplasty with piezoelectric osteotomies and mid-vault reconstruction. All structural cases were performed employing an open approach. Four revision surgeries were necessary. Open and closed approaches have indications depending on the tip and dorsal deformities. A closed PR is favored with thin skin, minimal dorsal modification, osseocartilaginous preservation (foundation techniques), less complex tip deformities, and overprojected noses. An open PR is favored for extensive dorsal modification, S-shaped nasal bones, complex tip deformities, and tip augmentation. Structural dorsal rhinoplasty is always conducted open and preferred for complex dorsal deformities and severe septal deviations.

First description of Eucoleus garfiai (Gallego and Mas-Coma, 1975) in wild boar (Sus scrofa) in Italy

Eucoleus garfiai (syn. Capillaria garfiai) is a nematode infecting lingual tissue of domestic and wild swine. Prevalence data for this parasite are scant and often related to accidental findings, occurring only in Japan and a few European countries. In this study, an epidemiological survey was performed in order to identify E. garfiai in wild boar from the Campania region, southern Italy. A total of 153 wild boar carcasses were inspected over the course of two hunting seasons (2019–2020). Histological examinations were performed on tongue samples fixed and stained with haematoxylin and eosin. The scraping of dorsal tongue tissue was carried out to collect adult worms for parasitological examination. Out of 153 wild boars, 40 (26.1%, 95% CI: 19.8–33.6%) tested positive for helminths and/or eggs in tongue tissues. Parasites were identified morphologically and identification was confirmed by molecular analysis of the 18S rRNA gene, showing a 99% nucleotide match with E. garfiai sequences available in literature. No statistically significant differences were found according to age, sex nor hunting province. Our findings agree with previous histopathological data confirming the low pathogenic impact of this nematode. The present study represents the first report of E. garfiai in wild boar from Italy.

In ovo injection of CHIR-99021 promotes feather follicles development via activating Wnt/β-catenin signaling pathway during chick embryonic period

The Wingless-types/beta-catenin (Wnt/β-catenin) signaling pathway plays an important role in embryonic development and affects the physiological development processes of feather follicles. To investigate the role of Wnt/β-catenin pathway in regulating feather follicles morphogenesis, in ovo injection of CHIR-99021, an activator of the Wnt/β-catenin signaling pathway, was conducted in chick embryo model. Initially, a total of 40 embryos were used to assess feather follicles morphogenesis and the expression of β-catenin (E9–E17). The histological results showed that feather follicle morphogenesis was mainly completed from E9 to E17. β-catenin was involved in the processing of the appearance of dermal cell condensation (E9) and the completion of the feather follicles morphogenesis (E17). Next, a total of 160 fertilized eggs were randomly divided into 8 groups for in ovo injection at E9, including a Normal Saline injected group (CON) and the 500, 1,000, 2,000, 5,000, 10,000, 50,000, and 100,000 ng CHIR-99021 groups. Dorsal skin tissue samples were collected at E17 for investigating feather follicles morphology and expressions of β-catenin and lymphoid enhancerbinding factor-1 (LEF1) at gene and protein levels. The results showed that feather follicle diameter in the injected groups were significantly (P < 0.05) increased with limit dose-independence compared to the CON group. CHIR-99021 significantly (P < 0.05) influenced the mRNA expressions of catenin beta-1 (CTNNB1) and downstream target LEF1. In ovo injection of CHIR-99021 caused that β-catenin and LEF1 were significantly (P < 0.05) increased followed the increased doses as determined by western blotting. The immunochemical results showed that β-catenin was detected in the dermal papilla of feather follicles. Given these results, this study suggests to developmental biology that in ovo injection of CHIR-99021 promoted feather follicles morphogenesis and development via activating Wnt/β-catenin signaling pathway and upregulating downstream target LEF1 during embryonic period in chick embryo model. Moreover, CHIR-99021 may be a strong candidate to promote the animal feather/hair industry, especially as a reference for bird feather production.

Analyzing immune response to engineered hydrogels by hierarchical clustering of inflammatory cell subsets.

Understanding the immune response to hydrogel implantation is critical for the design of immunomodulatory biomaterials. To study the progression of inflammation around poly(ethylene glycol) hydrogels presenting Arg-Gly-Asp (RGD) peptides and vascular endothelial growth factor, we used temporal analysis of high-dimensional flow cytometry data paired with intravital imaging, immunohistochemistry, and multiplexed proteomic profiling. RGD-presenting hydrogels created a reparative microenvironment promoting CD206+ cellular infiltration and revascularization in wounded dorsal skin tissue. Unbiased clustering algorithms (SPADE) revealed significant phenotypic transition shifts as a function of the cell-adhesion hydrogel properties. SPADE identified an intermediate macrophage subset functionally regulating in vivo cytokine secretion that was preferentially recruited for RGD-presenting hydrogels, whereas dendritic cell subsets were preferentially recruited to RDG-presenting hydrogels. Last, RGD-presenting hydrogels controlled macrophage functional cytokine secretion to direct polarization and vascularization. Our studies show that unbiased clustering of single-cell data provides unbiased insights into the underlying immune response to engineered materials.