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1.
Mycoses ; 67(8): e13774, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39092516

ABSTRACT

BACKGROUND: Fungi clinically relevant to human skin comprise prevalent commensals and well-known pathogens. Only rarely human skin harbours fungi that evade identification. OBJECTIVE: To characterise an enigmatic specimen isolated from a skin lesion. METHODS: A comprehensive clinical and mycological workup including conventional methods for phenotypic characterisation and sequencing based on internal transcribed spacer (ITS) and large subunit (LSU) regions to infer a phylogenetic tree. RESULTS: Cultures on common solid media were macroscopically inconspicuous initially until mycelial tufts developed on the surface, notably on potato dextrose agar. Polymorphous chlamydospores were detected but no aleurospores and ascomata. At 26°C, the isolate grew on standard agars, plant materials and garden soil and utilised peptone, keratins, lipids, inulin, erythrocytes and cellulose. It also grew at 5°C and at 37°C. Nucleotide sequences of its ITS region showed 93% similarity to sequences of different Malbranchea species. The closest matches among LSU rRNA sequences were obtained with the genera Amauroascus, Arthroderma, Auxarthronopsis and Malbranchea (93%-95%). A combined phylogenetic analysis placed the fungus in a sister clade to Neogymnomycetaceae, classified as incertae sedis in Onygenales, on a large distance to either Diploospora rosea or 'Amauroascus' aureus. CONCLUSIONS: The genus Inopinatus gen. nov. (MB854685) with the species Inopinatus corneliae sp. nov. (MB854687) is introduced to accommodate our isolate (holotype: DSM 116806; isotypes: CBS 151104, IHEM 29063). Probably Inopinatus corneliae is a geophilic species that, although potentially harmful, was no relevant pathogen in our case. Its ecology, epidemiology and pathogenicity need to be further clarified.


Subject(s)
DNA, Fungal , DNA, Ribosomal Spacer , Onygenales , Phylogeny , Sequence Analysis, DNA , Skin , Humans , Skin/microbiology , Onygenales/genetics , Onygenales/classification , Onygenales/isolation & purification , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics , Dermatomycoses/microbiology , Keratins/metabolism , DNA, Ribosomal/genetics , Male , Mycological Typing Techniques
2.
Zhonghua Bing Li Xue Za Zhi ; 53(8): 803-808, 2024 Aug 08.
Article in Chinese | MEDLINE | ID: mdl-39103261

ABSTRACT

Objective: To study the clinicopathological features, immunohistochemical phenotypes, molecular changes, differential diagnosis and prognosis of isolated intraductal carcinoma of the prostate (iIDC-P). Methods: Three iIDC-P cases were collected retrospectively from 2016 to 2022 at Ningbo Clinical Pathology Diagnosis Center, Ningbo, China. The clinicopathologic features and immunophenotypic profiles were studied using light microscopy and immunohistochemistry. A targeted next-generation sequencing panel was used to analyze cancer-associated mutations. Follow-up and literature review were also performed. Results: The patients' ages were 61, 67 and 77 years, and their preoperative prostate specific antigen (PSA) levels were 7.99, 7.99 and 4.86 µg/L, respectively. Case 1 and 2 were diagnosed on needle biopsy and radical prostatectomy (RP) specimens, and case 3 was diagnosed on a specimen of transurethral resection of the prostate (TURP). The RP specimen was entirely submitted for histologic examination. In the case 1, iIDC-P was found in one tissue core (involving two ducts) in the biopsy specimen, and in 6 sections (diameter, 0.3-1.1 cm) from the radical prostatectomy specimen, and one section had separate foci of low-grade acinar adenocarcinoma (diameter, 0.05 cm). In the case 2, 6 tissue sections from the biopsy specimens showed iIDC-P, and 13 sections from RP specimen showed iIDC-P (diameter, 0.5-1.6 cm), and the other 3 sections had separate low grade acinar adenocarcinoma (diameter, 0.6 cm). In the case 3, 5 tissue blocks from the TURP specimen showed iIDC-P. The case 1 and 2 showed solid architecture with expansile proliferation of neoplastic cells in native ducts and acini. The case 3 showed dense or loose cribriform pattern, with marked cytological atypia, and frequent mitotic figures. Comedonecrosis was found in solid or dense cribriform glands in the case 2. Immunohistochemically, surrounding basal cells were highlighted using high-molecular-weight cytokeratin (34ßE12 and CK5/6) and p63, while P504s was positive in the tumor cells. The tumor cells were also positive for AR and prostate markers (NKX3.1, PSA and PSAP), and negative for GATA3. The iIDC-P and acinar adenocarcinoma both showed weak PTEN expression and no ERG (nuclear) expression. In case 2 and 3, targeted sequencing revealed activated oncogenic driver mutations in MAPK and PI3K pathway genes (KRAS, MTOR and PTEN). In addition, pathogenic mutation in TP53 and FOXA1 mutation were found in the case 2 and 3, respectively. No case demonstrated TMPRSS2::ERG translocation. All cases were microsatellite stable and had lower tumor mutation burdens (range, 2.1-3.1 muts/Mb). The patients showed no biochemical recurrence or metastasis after follow-up of 16-91 months. Conclusions: iIDC-P is a special type of intraductal carcinoma of the prostate and differs from intraductal carcinoma within high-grade prostate cancer. iIDC-P has unique molecular characteristics and may represent as a molecularly unique in situ tumor of prostate cancer.


Subject(s)
Prostate-Specific Antigen , Prostatic Neoplasms , Humans , Male , Prostatic Neoplasms/pathology , Prostatic Neoplasms/genetics , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/surgery , Aged , Middle Aged , Retrospective Studies , Prostate-Specific Antigen/metabolism , Receptors, Androgen/metabolism , Receptors, Androgen/genetics , Diagnosis, Differential , Transcription Factors/metabolism , Transcription Factors/genetics , Carcinoma, Intraductal, Noninfiltrating/pathology , Carcinoma, Intraductal, Noninfiltrating/genetics , Carcinoma, Intraductal, Noninfiltrating/metabolism , Homeodomain Proteins/metabolism , Homeodomain Proteins/genetics , Carcinoma, Ductal/pathology , Carcinoma, Ductal/genetics , Carcinoma, Ductal/metabolism , Carcinoma, Ductal/surgery , Transurethral Resection of Prostate , Racemases and Epimerases/metabolism , Racemases and Epimerases/genetics , High-Throughput Nucleotide Sequencing , Prognosis , Keratins , Membrane Proteins
3.
Gene ; 927: 148751, 2024 Nov 15.
Article in English | MEDLINE | ID: mdl-38971547

ABSTRACT

By analyzing the expression patterns of inner root sheath (IRS) specific genes during different developmental stages of hair follicle (HF) in Tan sheep embryos and at birth, this study aims to reveal the influence of the IRS on crimped wool. Skin tissues from the scapular region of male Tan sheep were collected at 85 days (E85) and 120 days (E120) of fetal development, and at 0 days (D0), 35 days (D35), and 60 days (D60) after birth, with four samples at each stage. Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to determine the relative expression levels of IRS type I keratin genes (KRT25, KRT26, KRT27, KRT28), type II keratin genes (KRT71, KRT72, KRT73, KRT74), and the trichohyalin gene (TCHH) in the skin of Tan sheep at different stages. Results showed that the expression levels of all IRS-specific genes peaked at D0, with the expression of all genes significantly higher than at E85 (P < 0.01), except for KRT73 and TCHH. The expression levels of KRT25, KRT26, and KRT72 were also significantly higher than at E120 (P < 0.01). Furthermore, the expression levels of KRT27, KRT28, KRT71, and KRT74 were significantly higher than both at E120 and D35 (P < 0.01). The expression levels of other genes at different stages showed no significant difference (P > 0.05). Conclusion: The IRS-specific genes exhibit the highest expression levels in Tan sheep at the neonatal stage. The expression levels of KRT71, KRT72, and TCHH, which are consistent with the pattern of wool crimp, may influence the morphology of the IRS and thereby affect the crimp of Tan sheep wool.


Subject(s)
Gene Expression Regulation, Developmental , Hair Follicle , Animals , Hair Follicle/metabolism , Hair Follicle/growth & development , Sheep/genetics , Sheep/growth & development , Male , Wool/metabolism , Wool/growth & development , Keratins, Type II/genetics , Keratins, Type II/metabolism , Keratins/genetics , Keratins/metabolism , Keratins, Type I/genetics , Keratins, Type I/metabolism , Intermediate Filament Proteins
4.
Wounds ; 36(6): 183-188, 2024 06.
Article in English | MEDLINE | ID: mdl-39018361

ABSTRACT

BACKGROUND: Lower extremity wounds in patients with diabetes are difficult to heal due to an overabundance of pro-inflammatory M1 macrophages, reduced phagocytosis of necrosed cells, and circulatory issues. Keratin biomaterials have been shown to address some of these concerns by encouraging the proliferation of anti-inflammatory M2 macrophages, thereby creating more favorable conditions for wound healing resembling those of patients without diabetes. OBJECTIVE: To investigate the effect of a novel human keratin matrix (HKM) on wound healing. MATERIALS AND METHODS: Ten patients with diabetes with lower extremity wounds at risk for delayed healing underwent wound debridement and application of HKM. Patients received weekly follow-up care and reapplication of HKM until healing occurred; wound size at each visit was used to calculate healing rate. RESULTS: Increased healing rates were noted with HKM compared with standard of care (SOC), including debridement and collagen treatment in all 8 patients who had received SOC prior to HKM treatment. When HKM treatment was alternated with SOC in 2 patients due to other medical conditions, healing rates decreased with SOC and then increased after reintroduction of HKM applications. CONCLUSIONS: These results suggest that HKM may help regulate the pathological processes that contribute to wound chronicity to "kick-start" wound healing. This case series demonstrates that HKM is a promising technology to improve healing rates in nonhealing lower extremity wounds in patients with diabetes.


Subject(s)
Debridement , Diabetic Foot , Keratins , Wound Healing , Humans , Wound Healing/physiology , Wound Healing/drug effects , Male , Female , Diabetic Foot/therapy , Middle Aged , Aged , Debridement/methods , Keratins/metabolism , Treatment Outcome , Lower Extremity
5.
Medicina (Kaunas) ; 60(7)2024 Jul 17.
Article in English | MEDLINE | ID: mdl-39064586

ABSTRACT

Background and Objectives: The key factor that enables osteoblastic activity and the formation of new bone, as well as gingiva, during orthodontic tooth extrusion (OE) is the periodontal ligament. The reaction of periodontal tissues associated with changes in the gingiva is a part of orthodontic tooth displacement. The aim of this study was to examine the effect of OE on the width of the zone of the keratinized and attached gingiva, the position of the mucogingival junction, and the height of the interdental papillae in the region where the OE was performed as well as in the adjacent region. Materials and Methods: This research included 28 adult patients (both orthodontically treated and untreated). The treated group included 15 patients, in whom orthodontic extrusion of the upper or lower frontal teeth was indicated and performed. The untreated group included 13 patients, with no previous or undergoing orthodontic treatment. Patients with periodontal disease and periodontal pockets in the frontal region and patients allergic to iodine were excluded from the study. Gingivomorphometric measurements were performed on two occasions in three groups of teeth (24 extruded and 30 agonist teeth in the treated patients; 66 teeth in the untreated patients). Statistical analysis of the obtained data was performed using the software package SPSS version 26.0. Results: Orthodontic extrusion induced changes in the position of the mucogingival line and an increase in the width of the keratinized gingiva. There were no statistically significant effects on the depth of the gingival sulcus, the attached gingiva width, or the height of the interdental papillae. Conclusions: Orthodontic tooth extrusion has an effect on the periodontium in the observed region. Vertical orthodontic force, directed towards the coronal plane, affects the surrounding soft oral tissues.


Subject(s)
Gingiva , Orthodontic Extrusion , Humans , Female , Adult , Male , Orthodontic Extrusion/methods , Keratins , Young Adult , Periodontal Ligament
6.
Molecules ; 29(14)2024 Jul 18.
Article in English | MEDLINE | ID: mdl-39064958

ABSTRACT

The volume of difficult-to-process keratin waste is increasing as a result of rising global meat production. If not properly managed, this waste can contribute to environmental pollution and pose a threat to human and animal welfare. An interesting and more sustainable alternative is therefore the bioconversion of keratin using microorganisms and their enzymes. This work aimed to isolate bacteria from soil samples and zoonotic keratins and to evaluate their enzymatic capacity to degrade α- and ß-keratin wastes. A total of 113 bacterial strains were isolated from environmental samples and subjected to taxonomic identification using the MALDI-TOF MS technique and to a two-step screening for proteolytic and keratinolytic activity. The ability to degrade a ß-rich keratin substrate was observed in almost all of the strains isolated from soil and horsehairs. In contrast, when an α-rich keratin substrate was used, the highest levels of hydrolysis were observed only for Ker39, Ker66, Ker85, Ker100, and Ker101. Strains with the highest biodegradation potential were identified using molecular biology methods. Phylogenetic analysis of 16S rDNA gene sequences allowed the assignment of selected keratinolytic microorganisms to the genera Exiguobacterium, Priestia, Curtobacterium, Stenotrophomonas, Bacillus, Kocuria, or Pseudomonas. The results of this study are a promising precursor for the development of new, more sustainable methods of managing keratin waste to produce high-value hydrolysates.


Subject(s)
Bacteria , Biodegradation, Environmental , Keratins , Phylogeny , Keratins/metabolism , Bacteria/metabolism , Bacteria/genetics , RNA, Ribosomal, 16S/genetics , Soil Microbiology , Animals , Peptide Hydrolases/metabolism
7.
Cell Rep ; 43(7): 114480, 2024 Jul 23.
Article in English | MEDLINE | ID: mdl-39003737

ABSTRACT

The cytoskeleton of the cell is constantly exposed to physical forces that regulate cellular functions. Selected members of the LIM (Lin-11, Isl-1, and Mec-3) domain-containing protein family accumulate along force-bearing actin fibers, with evidence supporting that the LIM domain is solely responsible for this force-induced interaction. However, LIM domain's force-induced interactions are not limited to actin. LIMK1 and LMO1, both containing only two tandem LIM domains, are recruited to force-bearing keratin fibers in epithelial cells. This unique recruitment is mediated by their LIM domains and regulated by the sequences outside the LIM domains. Based on in vitro reconstitution of this interaction, LIMK1 and LMO1 directly interact with stretched keratin 8/18 fibers. These results show that LIM domain's mechano-sensing abilities extend to the keratin cytoskeleton, highlighting the diverse role of LIM proteins in force-regulated signaling.


Subject(s)
Intermediate Filaments , Keratins , LIM Domain Proteins , Lim Kinases , LIM Domain Proteins/metabolism , Humans , Lim Kinases/metabolism , Keratins/metabolism , Intermediate Filaments/metabolism , Protein Binding , Animals , Transcription Factors/metabolism
8.
Clin Exp Dent Res ; 10(4): e932, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38973200

ABSTRACT

OBJECTIVES: There is a growing evidence to suggest augmenting peri-implant keratinized mucosa in the presence of ≤ 2 mm of keratinized mucosa. However, the most appropriate surgical technique and augmentation materials have yet to be defined. The aim of this systematic review and meta-analyses was to evaluate the clinical and patient-reported outcomes of augmenting keratinized mucosa around implants using free gingival graft (FGG) versus xenogeneic collagen matrix (XCM) before commencing prosthetic implant treatment. MATERIAL AND METHODS: Electronic databases were searched to identify observational studies comparing implant sites augmented with FGG to those augmented with XCM. The risk of bias was assessed using the Cochrane Collaboration's Risk of Bias tool. RESULTS: Six studies with 174 participants were included in the present review. Of these, 87 participants had FGG, whereas the remaining participants had XCM. At 6 months, sites augmented with FGG were associated with less changes in the gained width of peri-implant keratinized mucosa compared to those augmented with XCM (mean difference 1.06; 95% confidence interval -0.01 to 2.13; p = 0.05). The difference, however, was marginally significant. The difference between the two groups in changes in thickness of peri-implant keratinized mucosa at 6 months was statistically significantly in favor of FGG. On the other hand, XCM had significantly shorter surgical time, lower postoperative pain score, and higher color match compared to FGG. CONCLUSIONS: Within the limitation of this review, the augmentation of keratinized mucosa using FGG before the placement of the final prosthesis may have short-term positive effects on soft tissue thickness. XCM might be considered in aesthetically demanding implant sites and where patient comfort or shorter surgical time is a priority. The evidence support, however, is of low to moderate certainty; therefore, further studies are needed to support the findings of the present review.


Subject(s)
Collagen , Dental Implants , Gingiva , Humans , Collagen/therapeutic use , Gingiva/transplantation , Gingiva/pathology , Gingiva/surgery , Keratins , Mouth Mucosa/transplantation , Gingivoplasty/methods , Dental Implantation, Endosseous/methods , Heterografts
9.
Int J Biol Macromol ; 275(Pt 2): 133722, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38977053

ABSTRACT

The valorization of discarded wool from dairy sheep breeding is a challenging issue. The most proposed strategies lie in the processing of keratin extracted from wool without reducing the molecular weight of the protein chains (the high molecular weight-HMW keratin). Here, the HMW keratin has been spun for the first time by solution blow spinning. A screening study of the process carried out with a 2-level full factorial design revealed that keratin filaments can be obtained by using the polyethylene oxide at 900 kDa, a 2 bar air pressure, and a 30 cm needle-collector distance. An annealing at 80 °C for 15 min, at pH 3.5 with citric acid contributes to increasing the viscosity of the keratin solutions thereby allowing the production of defect-free and water-stable filaments having diameters from 1 to 6 µm. A negligible toxic effect was observed after 24 and 48 h on HT29 epithelial cells and normal blood cells displayed behavior similar to the control demonstrating that the patches are hemocompatible. Therefore, the developed SBS process of keratin aqueous solutions could represent a valuable platform for developing patches that need to be blood-contacting and deposited in-situ.


Subject(s)
Keratins , Wool , Keratins/chemistry , Animals , Wool/chemistry , Sheep , Humans , Molecular Weight , Solutions
10.
Microb Physiol ; 34(1): 170-181, 2024.
Article in English | MEDLINE | ID: mdl-38955164

ABSTRACT

INTRODUCTION: The global poultry industry produces millions of tons of waste feathers every year, which can be bio-degraded to make feed, fertilizer, and daily chemicals. However, feather bio-degradation is a complex process that is not yet fully understood. This results in low degradation efficiency and difficulty in industrial applications. Omics-driven system biology research offers an effective solution to quickly and comprehensively understand the molecularmechanisms involved in a metabolic pathway. METHODS: In the early stage of this process, feathers are hydrolyzed into water-soluble keratin monomers. In this study, we used high-throughput RNA-seq technology to analyze the genes involved in the internalization and degradation of keratin monomers in Stenotrophomonas maltophilia DHHJ strain cells. Moreover, we used Co-IP with LC-MS/MS technology to search for proteins that interact with recombinant keratin monomers. RESULTS: We discovered TonB transports and molecular chaperones associating with the keratin monomer, which may play a crucial role in the transmembrane transport of keratin. Meanwhile, multiple proteases belonging to distinct families were identified as binding partners of keratin monomers, among which ATPases associated with diverse cellular activity (AAA+) family proteases are overrepresented. Four genes, including JJL50_15620, JJL50_17955 (TonB-dependent receptors), JJL50_03260 (ABC transporter ATP-binding protein), and JJL50_20035 (ABC transporter substrate-binding protein), were selected as representatives for determining their expressions under different culture conditions using qRT-PCR, and they were found to be upregulated in response to keratin degradation consistent with the data from RNA-seq and Co-IP. CONCLUSION: This study highlights the complexity of keratin biodegradation in S. maltophilia DHHJ, in which multiple pathways are involved such as protein folding, protein transport, and several protease systems. Our findings provide new insights into the mechanism of feather degradation.


Subject(s)
Bacterial Proteins , Biodegradation, Environmental , Keratins , Stenotrophomonas maltophilia , Stenotrophomonas maltophilia/metabolism , Stenotrophomonas maltophilia/genetics , Keratins/metabolism , Keratins/genetics , Animals , Bacterial Proteins/metabolism , Bacterial Proteins/genetics , Feathers/metabolism , Feathers/microbiology , Tandem Mass Spectrometry , Gene Expression Regulation, Bacterial , Peptide Hydrolases/metabolism , Peptide Hydrolases/genetics
11.
Int J Pharm ; 661: 124451, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-38992735

ABSTRACT

We extended a mechanistic, physics-based framework of the dry down process, previously developed for liquids and electrolytes, to solids and coded it into the latest UB/UC/P&G skin permeation model, herein renamed DigiSkin. The framework accounts for the phase change of the permeant from dissolved in a solvent (liquid) to precipitated on the skin surface (solid). The evaporation rate for the solid is reduced due to lower vapor pressure for the solid state versus subcooled liquid. These vapor pressures may differ by two orders of magnitude. The solid may gradually redissolve and penetrate the skin. The framework was tested by simulating the in vitro human skin permeation of the 38 cosmetically relevant solid compounds reported by Hewitt et al., J. Appl. Toxicol. 2019, 1-13. The more detailed handling of the evaporation process greatly improved DigiSkin evaporation predictions (r2 = 0.89). Further, we developed a model reliability prediction score classification using diverse protein reactivity data and identified that 15 of 38 compounds are out of model scope. Dermal delivery predictions for the remaining chemicals have excellent agreement with experimental data. The analysis highlighted the sensitivity of water solubility and equilibrium vapor pressure values on the DigiSkin predictions outcomes influencing agreement with the experimental observations.


Subject(s)
Cosmetics , Keratins , Skin Absorption , Skin , Solvents , Solvents/chemistry , Humans , Hydrogen-Ion Concentration , Skin/metabolism , Keratins/chemistry , Cosmetics/chemistry , Cosmetics/pharmacokinetics , Administration, Cutaneous , Solubility , Models, Biological , Pharmaceutical Vehicles/chemistry , Phase Transition
12.
Nat Commun ; 15(1): 6259, 2024 Jul 24.
Article in English | MEDLINE | ID: mdl-39048559

ABSTRACT

Pityriasis rubra pilaris (PRP) is an inflammatory papulosquamous dermatosis, characterized by hyperkeratotic follicular papules and erythematous desquamative plaques. The precise pathogenic mechanism underlying PRP remains incompletely understood. Herein, we conduct a case-control study involving a cohort of 102 patients with sporadic PRP and 800 healthy controls of Han Chinese population and identify significant associations (P = 1.73 × 10-6) between PRP and heterozygous mutations in the Keratin 32 gene (KRT32). KRT32 is found to be predominantly localized in basal keratinocytes and exhibits an inhibitory effect on skin inflammation by antagonizing the NF-κB pathway. Mechanistically, KRT32 binds to NEMO, promoting excessive K48-linked polyubiquitination and NEMO degradation, which hinders IKK complex formation. Conversely, loss-of-function mutations in KRT32 among PRP patients result in NF-κB hyperactivation. Importantly, Krt32 knockout mice exhibit a PRP-like dermatitis phenotype, suggesting compromised anti-inflammatory function of keratinocytes in response to external pro-inflammatory stimuli. This study proposes a role for KRT32 in regulating inflammatory immune responses, with damaging variants in KRT32 being an important driver in PRP development. These findings offer insights into the regulation of skin immune homeostasis by keratin and open up the possibility of using KRT32 as a therapeutic target for PRP.


Subject(s)
Keratinocytes , Pityriasis Rubra Pilaris , Skin , Adult , Animals , Female , Humans , Male , Mice , Middle Aged , Young Adult , Case-Control Studies , Homeostasis , I-kappa B Kinase/genetics , I-kappa B Kinase/metabolism , Keratinocytes/immunology , Keratinocytes/metabolism , Keratins/metabolism , Keratins/genetics , Loss of Function Mutation , Mice, Knockout , NF-kappa B/metabolism , Pityriasis Rubra Pilaris/genetics , Pityriasis Rubra Pilaris/immunology , Pityriasis Rubra Pilaris/pathology , Pityriasis Rubra Pilaris/metabolism , Signal Transduction , Skin/pathology , Skin/immunology , Skin/metabolism , Ubiquitination
13.
J Environ Manage ; 366: 121648, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39018864

ABSTRACT

The purpose of this work is to produce keratin hydrolysate from sheep wool by alkaline hydrolysis and to assess its effectiveness in improving maize plant growth under greenhouse conditions. A hybrid response surface methodology with Box-Behnken design (RSM-BBD) was used to model and optimize the hydrolysis process. The synergistic effects between three critical independent variables including temperature, hydrolysis time, and concentration of KOH on the hydrolysis rate were statistically investigated and optimized. Under optimized conditions, a hydrolysis rate of 95.08% was achieved. The produced hydrolysate consists of water-soluble peptides, free amino acids and potassium ions, making it suitable to be used as a valuable agricultural input material for crop production. Amino acid analysis revealed high levels of proline and phenylalanine, which are responsible for water conditioning and the preservation of abiotic stress as readily available. The efficacy of the produced hydrolysate was assessed in the cultivation of maize as a crop model under greenhouse conditions. Results revealed that the application of the hydrolysate positively influenced the morphological traits of the maize crop such as plant height and leaf surface area. The magnitude of the response to the hydrolysate application depended on its concentration with the most positive effects observed at a dose 2 for the leaf's chlorophyll content, fresh shoot biomass and dry shoot biomass. The application of the hydrolysate improved fresh and dry shoot biomass by 32.5 and 34.4% compared to the control and contributed to the improvement of nitrogen use efficiency by the studied crop. The hydrolysate proved to be beneficial in improving overall plant growth and can be suitable and effective agricultural input for maize cultivation.


Subject(s)
Keratins , Wool , Zea mays , Zea mays/growth & development , Zea mays/metabolism , Animals , Wool/growth & development , Keratins/metabolism , Sheep , Hydrolysis , Biomass , Amino Acids/metabolism
14.
Int J Biol Macromol ; 275(Pt 2): 133690, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38971280

ABSTRACT

In pursuing sustainable thermal insulation solutions, this study explores the integration of human hair and feather keratin with alginate. The aim is to assess its potential in thermal insulation materials, focusing on the resultant composites' thermal and mechanical characteristics. The investigation uncovers that the type and proportion of keratin significantly influence the composites' porosity and thermal conductivity. Specifically, higher feather keratin content is associated with lesser sulfur and reduced crosslinking due to shorter amino acids, leading to increased porosity and pore sizes. This, in turn, results in a decrease in ß-structured hydrogen bond networks, raising non-ordered protein structures and diminishing thermal conductivity from 0.044 W/(m·K) for pure alginate matrices to between 0.033 and 0.038 W/(m·K) for keratin-alginate composites, contingent upon the specific ratio of feather to hair keratin used. Mechanical evaluations further indicate that composites with a higher ratio of hair keratin exhibit an enhanced compressive modulus, ranging from 60 to 77 kPa, demonstrating the potential for tailored mechanical properties to suit various applications. The research underscores the critical role of sulfur content and the crosslinking index within keratin's structures, significantly impacting the thermal and mechanical properties of the matrices. The findings position keratin-based composites as environmentally friendly alternatives to traditional insulation materials.


Subject(s)
Feathers , Hair , Keratins , Thermal Conductivity , Keratins/chemistry , Feathers/chemistry , Hair/chemistry , Humans , Alginates/chemistry , Porosity
15.
Int J Biol Macromol ; 276(Pt 2): 133805, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38996885

ABSTRACT

Successful oral insulin administration can considerably enhance the quality of life (QOL) of diabetes patients who must frequently take insulin injections. However, Oral insulin administration is seriously hampered by gastrointestinal enzymes, wide pH range, mucus and mucosal layers, which limit insulin oral bioavailability to ≤2 %. Herein, we developed a simple, inexpensive and safe dual ß-cyclodextrin/dialdehyde glucan-coated keratin nanoparticle (ß-CD-K-IN-DG). The resulted ß-CD-K-IN-DG not only gave the ultra-high insulin loading (encapsulation efficiency (98.52 %)), but also protected insulin from acid and enzymatic degradation. This ß-CD-K-IN-DG had a notable hypoglycemic effect, there was almost 80 % insulin release after 4 h of incubation under hyperglycemic conditions. Ex vivo results confirmed that ß-CD-K-IN-DG possessed high mucus-penetration ability. Transepithelial transport and uptake mechanism studies revealed that bypass transport pathway and endocytosis promoted ß-CD-K-IN-DG entered intestinal epithelial cells, thus increased the bioavailability of insulin (12.27 %). The improved stability of insulin during in vivo transport implied that ß-CD-K-IN-DG might be a potential tool for the effective oral insulin administration.


Subject(s)
Drug Carriers , Insulin , Keratins , Nanoparticles , beta-Cyclodextrins , Nanoparticles/chemistry , Insulin/administration & dosage , Insulin/pharmacokinetics , Insulin/chemistry , beta-Cyclodextrins/chemistry , Administration, Oral , Humans , Animals , Keratins/chemistry , Drug Carriers/chemistry , Glucans/chemistry , Biological Availability , Hypoglycemic Agents/administration & dosage , Hypoglycemic Agents/pharmacokinetics , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/pharmacology , Mice , Male , Caco-2 Cells , Drug Liberation
16.
Int J Pharm ; 661: 124372, 2024 Aug 15.
Article in English | MEDLINE | ID: mdl-38909923

ABSTRACT

Free amino acids (FAAs) constitute the largest component (∼40 %) of the so-called natural moisturizing factors of the skin. Their level declines in dry skin conditions and one strategy to overcome this problem may involve the topical delivery of FAAs through appropriate strategy. The objective of the present study was therefore to identify alternative skin models and study the corneocyte-water partition coefficients (KCOR/W) and permeation coefficient (KP) of 18 FAAs. The KCOR/W was studied using standard protocols and the permeation studies were conducted using Franz diffusion cell. The results indicate that the FAAs have high partitioning behavior to the corneocytes. The KCOR/W values of the human COR and that of pig ear skin were better correlated with each other than that of keratin isolated from chicken feathers. The presence of lipid in the stratum corneum (SC), initial concentration of the FAAs, and permeation enhancers affect the KCOR/W. The FAAs have low permeation into the SC which suggests the need for permeation enhancers in designing dosage form containing these compounds. Even though the investigated mathematical models show good prediction of the Kp values, better prediction could be obtained by considering factors such as the possible entrapment of the FAAs by the CORs.


Subject(s)
Amino Acids , Hydrophobic and Hydrophilic Interactions , Permeability , Skin Absorption , Animals , Amino Acids/chemistry , Amino Acids/metabolism , Amino Acids/pharmacokinetics , Swine , Humans , Skin/metabolism , Administration, Cutaneous , Chickens , Epidermis/metabolism , Models, Biological , Keratins/metabolism , Keratins/administration & dosage
17.
Cell Death Dis ; 15(6): 439, 2024 Jun 21.
Article in English | MEDLINE | ID: mdl-38906852

ABSTRACT

Although adamantinomatous craniopharyngioma (ACP) is a tumour with low histological malignancy, there are very few therapeutic options other than surgery. ACP has high histological complexity, and the unique features of the immunological microenvironment within ACP remain elusive. Further elucidation of the tumour microenvironment is particularly important to expand our knowledge of potential therapeutic targets. Here, we performed integrative analysis of 58,081 nuclei through single-nucleus RNA sequencing and spatial transcriptomics on ACP specimens to characterize the features and intercellular network within the microenvironment. The ACP environment is highly immunosuppressive with low levels of T-cell infiltration/cytotoxicity. Moreover, tumour-associated macrophages (TAMs), which originate from distinct sources, highly infiltrate the microenvironment. Using spatial transcriptomic data, we observed one kind of non-microglial derived TAM that highly expressed GPNMB close to the terminally differentiated epithelial cell characterized by RHCG, and this colocalization was verified by asmFISH. We also found the positive correlation of infiltration between these two cell types in datasets with larger cohort. According to intercellular communication analysis, we report a regulatory network that could facilitate the keratinization of RHCG+ epithelial cells, eventually causing tumour progression. Our findings provide a comprehensive analysis of the ACP immune microenvironment and reveal a potential therapeutic strategy base on interfering with these two types of cells.


Subject(s)
Craniopharyngioma , Pituitary Neoplasms , Tumor Microenvironment , Humans , Craniopharyngioma/genetics , Craniopharyngioma/pathology , Craniopharyngioma/metabolism , Craniopharyngioma/immunology , Tumor Microenvironment/immunology , Pituitary Neoplasms/pathology , Pituitary Neoplasms/genetics , Pituitary Neoplasms/immunology , Pituitary Neoplasms/metabolism , Tumor-Associated Macrophages/metabolism , Tumor-Associated Macrophages/immunology , Male , Female , Keratins/metabolism , Transcriptome/genetics , Gene Expression Regulation, Neoplastic , Adult , Middle Aged , Multiomics
18.
Sci Rep ; 14(1): 12864, 2024 06 04.
Article in English | MEDLINE | ID: mdl-38834664

ABSTRACT

Natural polymer-based hydrogels have demonstrated great potential as wound-healing dressings. They help to maintain a moist wound environment as well as promote faster healing. In this work, a multifunctional hydrogel was prepared using keratin, sodium alginate, and carboxymethyl chitosan with tannic acid modification. Micro-morphology of hydrogels has been performed by scanning electron microscopy. Fourier Transform Infrared Spectroscopy reveals the presence of hydrogen bonding. The mechanical properties of the hydrogels were examined using a universal testing machine. Furthermore, we investigated several properties of the modified hydrogel. These properties include swelling rate, water retention, anti-freezing properties, antimicrobial and antioxidant properties, hemocompatibility evaluation and cell viability test in vitro. The modified hydrogel has a three-dimensional microporous structure, the swelling rate was 1541.7%, the elastic modulus was 589.74 kPa, the toughness was 211.74 kJ/m3, and the elongation at break was 75.39%, which was similar to the human skin modulus. The modified hydrogel also showed inhibition of S. aureus and E. coli, as well as a DPPH scavenging rate of 95%. In addition, the modified hydrogels have good biological characteristics. Based on these findings, the K/SA/CCS hydrogel holds promise for applications in biomedical engineering.


Subject(s)
Alginates , Chitosan , Hydrogels , Keratins , Tannins , Alginates/chemistry , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antioxidants/chemistry , Antioxidants/pharmacology , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Cell Survival/drug effects , Chitosan/chemistry , Chitosan/analogs & derivatives , Elastic Modulus , Escherichia coli/drug effects , Hydrogels/chemistry , Keratins/chemistry , Spectroscopy, Fourier Transform Infrared , Staphylococcus aureus/drug effects , Tannins/chemistry
20.
Curr Microbiol ; 81(8): 227, 2024 Jun 16.
Article in English | MEDLINE | ID: mdl-38879855

ABSTRACT

Microbial degradation of keratin is characterized by its inherent safety, remarkable efficiency, and the production of copious degradation products. All these attributes contribute to the effective management of waste materials at high value-added and in a sustainable manner. Microbial degradation of keratin materials remains unclear, however, with variations observed in the degradation genes and pathways among different microorganisms. In this study, we sequenced the transcriptome of Purpureocillium lilacinum GZAC18-2JMP mycelia on control medium and the medium containing 1% feather powder, analyzed the differentially expressed genes, and revealed the degradation mechanism of chicken feathers by P. lilacinum GZAC18-2JMP. The results showed that the chicken feather degradation rate of P. lilacinum GZAC18-2JMP reached 64% after 216 h of incubation in the fermentation medium, reaching a peak value of 148.9 µg·mL-1 at 192 h, and the keratinase enzyme activity reached a peak value of 211 U·mL-1 at 168 h, which revealed that P. lilacinum GZAC18-2JMP had a better keratin degradation effect. A total of 1001 differentially expressed genes (DEGs) were identified from the transcriptome database, including 475 upregulated genes and 577 downregulated genes. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis of the DEGs revealed that the metabolic pathways related to keratin degradation were mainly sulfur metabolism, ABC transporters, and amino acid metabolism. Therefore, the results of this study provide an opportunity to gain further insight into keratin degradation and promote the biotransformation of feather wastes.


Subject(s)
Feathers , Hypocreales , Keratins , Transcriptome , Keratins/metabolism , Hypocreales/genetics , Hypocreales/metabolism , Animals , Feathers/metabolism , Chickens , Gene Expression Profiling , Fungal Proteins/genetics , Fungal Proteins/metabolism , Peptide Hydrolases/metabolism , Peptide Hydrolases/genetics , Mycelium/genetics , Mycelium/metabolism , Mycelium/growth & development , Fermentation , Biodegradation, Environmental
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