Search

In our previous works, two techniques (freeze-drying and salt-leaching) were introduced to fabricate the sericin/poly(vinyl alcohol)/glycerin scaffolds. The freeze-dried and salt-leached sericin/poly(vinyl alcohol)/glycerin scaffolds with the same composition showed distinguished physical and in vitro biological characteristics. In this study, the in vivo safety and efficacy tests of both scaffolds as dressing materials for the healing of full-thickness wounds in rat model were performed in comparison with the clinically used dressing, Allevyn[®]. In the safety test, the scaffolds were implanted subcutaneously, and the signs of tissue irritation including the extent of inflammatory cells, calcification, vascularization, and fatty infiltration were scored. In the efficacy test, the scaffolds were applied to the full-thickness wound (1.5 cm × 1.5 cm), and the epithelialization and collagen formation in the wound were evaluated. Both freeze-dried and salt-leached scaffolds were characterized as non- to slightly irritant implantable materials. The freeze-dried scaffold minimally causes irritation to the tissue possibly because it was derived from the non-chemical relevant process. Furthermore, the freeze-dried scaffold showed the highest wound healing efficiency as characterized by the fastest epithelialization and highest extent of collagen formation. This might be due to the more sustained release of sericin from the freeze-dried scaffold, compared to that of the salt-leached scaffold. Therefore, fabrication process seemed to directly regulate the properties and applicability of the scaffolds. The safety and efficacy of the dressing materials in wound healing application depended not only on the materials themselves but also on the fabrication process that produces them. [ABSTRACT FROM AUTHOR]

Phikud Navakot (PN), Thai herbal remedy in National List of Essential Medicines, has been claimed to reduce many cardiovascular symptoms especially dizziness and fainting. Apart from blood supply, erythrocyte morphology, in both shape and size, is one of the main consideration factors in cardiovascular diseases and may be affected by vascular oxidative stress. However, little is known about antioxidative property of PN on erythrocyte to preserve red blood cell integrity. In this study, 1,000 μM hydrogen peroxide-induced oxidative stress was conducted on sheep erythrocyte. Three doses of PN (1, 0.5, and 0.25 mg/mL) and 10 μM of ascorbic acid were compared. The released hemoglobin absorbance was measured to demonstrate hemolysis. Electron microscopic and immunohistochemical studies were also performed to characterize dysmorphic erythrocyte and osmotic ability in relation to aquaporin- (AQP-) 1 expression, respectively. The results revealed that all doses of PN and ascorbic acid decreased the severity of dysmorphic erythrocyte, particularly echinocyte, acanthocyte, knizocyte, codocyte, clumping, and other malformations. However, the most effective was 0.5 mg/mL PN dosage. In addition, hydrostatic pressure may be increased in dysmorphic erythrocyte in association with AQP-1 upregulation. Our results demonstrated that PN composes antioxidative effect to maintain the integrity and osmotic ability on sheep erythrocyte. [ABSTRACT FROM AUTHOR]

Simple Summary: Aedes aegypti (Ae. aegypti) is a mosquito that transmits arboviruses and responds to various biological and environmental stressors, including temperature, rainfall, and humidity. However, there is a lack of knowledge about fine particulate matter (PM2.5) effects on arbovirus vectors. We hypothesized that fine particulate matter (PM2.5) may affect Ae. aegypti blood-feeding rate and organs. We set up an environmental chamber that could adjust the concentration of PM2.5 and observed their blood-feeding activity. We observed a dose–response relationship between PM2.5 level and blood-feeding rate in adult female Ae. aegypti mosquitoes. In addition, histopathological study showed some changes in the ovaries. Vacuolated or vacuolar degeneration characterized by a formation of non-lipid small droplets in the cytoplasm was observed. This demonstrated the degenerative stage of the cells before developing hydropic degeneration or another advanced stage of cellular damage. The present study explored the effects of PM2.5 exposure on the blood-feeding rate and organ integrity in the major arboviral vector Ae. Aegypti, providing information on the potential but indirect operational impact of PM2.5 exposure on the survival and transmission capabilities of this major vector. Our findings may contribute towards the conceptualization and implementation of mosquito control measures with due consideration of the effects of ambient PM2.5 on their populations. High levels of fine particulate matter (PM2.5) air pollution are a concern for human health. Several studies have examined the effects of air pollution on human and animal health. However, there is a lack of knowledge about its effects on arbovirus vectors. Thus, we investigated whether PM2.5 concentration alters the blood-feeding activity of Ae. aegypti mosquitoes. We investigated the effect on the females' propensity to blood feed at eight concentrations of PM2.5 ranging from 100 to 1000 μg/m3. Correlation analysis showed blood-feeding activity had a significant strong negative correlation with concentration of PM2.5 (rp = −0.85; p ≤ 0.00001). Exploratory linear and non-linear models showed an exponential decay relationship was the best fitting model (corrected Akaike's information criterion, 193.0; Akaike's weight, 0.766; adjusted R2, 0.780). Ultrastructural study demonstrated PM2.5 did not obstruct the respiratory system, but some fine particles were present on the antenna and abdominal body parts. Ovaries showed a dose–response relationship between PM2.5 level and vacuolated degeneration. In conclusion, the blood-feeding behavior of Ae. aegypti females may have an exponential decay relationship with PM2.5 level, and their ovaries may demonstrate dose-dependent degeneration. These findings may be important in understanding the vector's biology and disease transmission in settings with high PM2.5 levels. These results are important to understand blood-feeding and feeding pattern of mosquitoes during PM2.5 pollution, which is important for disease transmission and vector control. [ABSTRACT FROM AUTHOR]

Therapeutic treatment forms can play significant roles in resolving psoriatic plaques or promoting wound repair in psoriatic skin. Considering the biocompatibility, mechanical strength, flexibility, and adhesive properties of silk fibroin sheets/films, it is useful to combine them with anti-psoriatic agents and healing stimulants, notably silk sericin. Here, we evaluate the curative properties of sericin-coated thin polymeric films (ScF) fabricated from silk fibroin, using an imiquimod-induced psoriasis rat model. The film biocompatibility and psoriatic wound improvement capacity was assessed. A proteomics study was performed to understand the disease resolving mechanisms. Skin-implantation study exhibited the non-irritation property of ScF films, which alleviate eczema histopathology. Immunohistochemical and gene expression revealed the depletion of β-defensin, caspase-3 and -9, TNF-α, CCL-20, IL-1β, IL-17, TGF-β, and Wnt expressions and S100a14 mRNA level. The proteomics study suggested that ScF diminish keratinocyte proliferation via the mTOR pathway by downregulating mTOR protein, corresponding to the modulation of TNF-α, Wnt, and IL-1β levels, leading to the enhancement of anti-inflammatory environment by IL-17 downregulation. Hematology data demonstrated the safety of using these biomaterials, which provide a potential therapeutic-option for psoriasis treatment due to desirable effects, especially anti-proliferation and anti-inflammation, functioning via the mTOR pathway and control of IL-17 signaling. [ABSTRACT FROM AUTHOR]

Lodderomyces elongisporus, first isolated in 1952, has increasingly been recognized as a significant pathogen, with a notable rise in human infections since the 1970s. Initially misidentified as Candida parapsilosis due to morphological similarities, L. elongisporus has now been conclusively established as a distinct species, largely due to advancements in molecular biology, particularly DNA sequencing. This review traces the detection history of L. elongisporus, from the earliest documented cases to the most recent reports, underscoring its role as a causative agent in human infections. It also explores therapeutic strategies that have demonstrated efficacy, alongside instances of environmental contamination reported in international literature. A critical evaluation of diagnostic methodologies essential for precise identification is provided, including culture-based techniques such as colony morphology on Sabouraud Dextrose Agar (SDA) and chromogenic media, coupled with microscopic assessments using Lactophenol Cotton Blue (LPCB) and Gram staining. The ultrastructure of L. elongisporus, as observed under Scanning Electron Microscopy (SEM), is also discussed. Furthermore, non-culture-based diagnostics, such as sugar utilization tests (API 20C AUX and the innovative in-house arabinose-based "Loddy" test) and antifungal susceptibility profiling, are reviewed, with a particular focus on molecular tools like ITS-DNA sequencing and MALDI-TOF MS, which, despite their higher costs, offer unparalleled specificity. The accurate distinction and characterization of L. elongisporus are paramount, particularly in vulnerable and immunocompromised patients, where misdiagnosis can lead to severe consequences. This review advocates for intensified research efforts to develop more accessible diagnostic tools and deepen our understanding of this emerging pathogen, ultimately aiming to improve patient outcomes., (© 2024. The Author(s).)

Gnathostomiasis, caused by the advanced third-stage larvae of Gnathostoma spinigerum , demands novel treatment avenues. The ethanolic root extract of Stemona collinsiae has been postulated to have anthelminthic properties, suggesting its potential as an alternative remedy. In this study, S. collinsiae roots were collected, identified, and extracted with 95 % ethanol. The crude extracts were standardized using didehydrostemofoline as chemical marker. The efficacy of the S. collinsiae root extract against third-stage larvae of G. spinigerum and its toxicity to Wistar rats were evaluated. Both in vitro and in vivo tests were performed, where the in vitro tests assessed the anthelminthic potential of S. collinsiae extract against G. spinigerum larvae, while in vivo tests examined the extract's efficacy against G. spinigerum larvae in infected Wistar rats and the efficacy was compared with albendazole. Parallelly, Wistar rats underwent acute and sub-chronic toxicity tests to establish the safe dosage of the extract. The in vitro tests showcased significant anthelminthic activity, marked by discernible morphological alterations in the exposed larvae. Acute toxicity proved fatal at 2000 mg/kg body weight, while a dose of 300 mg/kg proved non-toxic. Using the Globally Harmonized Classification System, an LD50 of 500 mg/kg was determined. In vivo trials revealed a pronounced decline in G. spinigerum larvae among rats treated with the S. collinsiae extract. The larvae were also observed to be encysted post-treatment, while those treated with albendazole were not encysted. The S. collinsiae extract, with its noteworthy in vitro efficacy and favorable safety metrics in rodents, can be a potential anthelminthic agent. The diminished inflammatory response compared to albendazole hints at S. collinsiae being a safer gnathostomiasis treatment alternative. The promising results in these preliminary trials warrant a deeper investigation to determine the root extract's optimal dosing, suitable delivery methods, and its broader clinical implications., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

The quorum sensing (QS) system mediated by the abaI gene in Acinetobacter baumannii is crucial for various physiological and pathogenic processes. In this study, we constructed a stable markerless abaI knockout mutant (ΔabaI) strain using a pEXKm5-based allele replacement method to investigate the impact of abaI on A. baumannii. Proteomic analysis revealed significant alterations in protein expression between the wild type (WT) and ΔabaI mutant strains, particularly in proteins associated with membrane structure, antibiotic resistance, and virulence. Notably, the downregulation of key outer membrane proteins such as SurA, OmpA, OmpW, and BamA suggests potential vulnerabilities in outer membrane integrity, which correlate with structural abnormalities in the ΔabaI mutant strain, including irregular cell shapes and compromised membrane integrity, observed by scanning and transmission electron microscopy. Furthermore, diminished expression of regulatory proteins such as OmpR and GacA-GacS highlights the broader regulatory networks affected by abaI deletion. Functional assays revealed impaired biofilm formation and surface-associated motility in the mutant strain, indicative of altered colonization capabilities. Interestingly, the mutant showed a complex antibiotic susceptibility profile. While it demonstrated increased susceptibility to membrane-targeting antibiotics, its response to beta-lactams was more nuanced. Despite increased expression of metallo-beta-lactamase (MBL) superfamily proteins and DcaP-like protein, the mutant unexpectedly showed lower MICs for carbapenems (imipenem and meropenem) compared to the wild-type strain. This suggests that abaI deletion affects antibiotic susceptibility through multiple, potentially competing mechanisms. Further investigation is needed to fully elucidate the interplay between quorum sensing, antibiotic resistance genes, and overall antibiotic susceptibility in A. baumannii. Our findings underscore the multifaceted role of the abaI gene in modulating various cellular processes and highlight its significance in A. baumannii physiology, pathogenesis, and antibiotic resistance. Targeting the abaI QS system may offer novel therapeutic strategies for this clinically significant pathogen., (© 2024. The Author(s).)

Giardia duodenalis, a widespread parasitic flagellate protozoan causing giardiasis, affects millions annually, particularly impacting children and travellers. With no effective vaccine available, treatment primarily relies on the oral administration of drugs targeting trophozoites in the small intestine. However, existing medications pose challenges due to side effects and drug resistance, necessitating the exploration of novel therapeutic options. Isocryptolepine, derived from Cryptolepis sanguinolenta, has demonstrated promising antimicrobial and anticancer properties. This study evaluated eighteen isocryptolepine-triazole adducts for their antigiardial activities and cytotoxicity, with ISO2 demonstrating potent antigiardial activity and minimal cytotoxicity on human intestinal cells. Metabolomics analysis revealed significant alterations in G. duodenalis metabolism upon ISO2 treatment, particularly affecting phospholipid metabolism. Notably, the upregulation of phytosphingosine and triglycerides, and downregulation of certain fatty acids, suggest a profound impact on membrane composition and integrity, potentially contributing to the parasite's demise. Pathway analysis highlighted glycerophospholipid metabolism, cytochrome b5 family heme/steroid binding domain, and P-type ATPase mechanisms as critical pathways affected by ISO2 treatment, underscoring its importance as a potential target for antigiardial therapy. These findings shed light on the mode of action of ISO2 against G. duodenalis and provide valuable insights for further drug development. Moreover, the study also offers a promising avenue for the exploration of isocryptolepine derivatives as novel therapeutic agents for giardiasis, addressing the urgent need for more effective and safer treatment options., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Stemona collinsiae Craib., Stemonaceae, has been traditionally used as medicinal plants for insecticides, treatment of parasitic worms and various diseases in Southeast Asian countries. Its ethanolic root extract has been postulated for anthelminthic activities which has a potential for development for human gnathostomiasis drug. To investigate the pharmacokinetic profile, liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method for the quantification of didehydrostemofoline in rats' plasma was developed and validated. The chromatographic separation was performed on a C 18 column using 1 mM ammonium acetate in water and methanol (50:50, v/v). Tetrahydropalmatine was used as an internal standard. The multiple reaction monitoring mode was used for quantitative analysis. The validated method showed good sensitivity, linearity, precision, and accuracy. The results of stability showed that didehydrostemofoline was stable in the extracted samples in auto-sampler for 24 h and in the plasma samples under room temperature for 24 h, -20 °C for 1 month, and after three freeze-thaw processes. The developed method was applied to the pharmacokinetic study of didehydrostemofoline after oral administration of S. collinsiae root extract. Didehydrostemofoline was rapidly absorbed from the gastrointestinal tract. The time to peak drug concentration was 1.75 ± 0.62 h with maximum drug concentration of 1152.58 ± 271.18 ng/mL. Didehydrostemofoline was rapidly eliminated from the body with terminal half-life of 1.86 ± 0.50 h. Calculated drug clearance of didehydrostemofoline was 96.82 ± 23.51 L/h and volume of distribution was 260.40 ± 96.81 L. The present study provided useful data for understanding drug disposition in the body with dynamic time-course which could be beneficial for further clinical trials., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could appear to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Schistosomiasis, caused by Schistosoma trematodes, is a significant global health concern, particularly affecting millions in Africa and Southeast Asia. Despite efforts to combat it, the rise of praziquantel (PZQ) resistance underscores the need for new treatment options. Protein kinases (PKs) are vital in cellular signaling and offer potential as drug targets. This study focused on focal adhesion kinase (FAK) as a candidate for anti-schistosomal therapy. Transcriptomic and proteomic analyses of adult S. mekongi worms identified FAK as a promising target due to its upregulation and essential role in cellular processes. Molecular docking simulations assessed the binding energy of FAK inhibitors to Schistosoma FAK versus human FAK. FAK inhibitor 14 and PF-03814735 exhibited strong binding to Schistosoma FAK with minimal binding for human FAK. In vitro assays confirmed significant anti-parasitic activity against S. mekongi, S. mansoni, and S. japonicum, comparable to PZQ, with low toxicity in human cells, indicating potential safety. These findings highlight FAK as a promising target for novel anti-schistosomal therapies. However, further research, including in vivo studies, is necessary to validate efficacy and safety before clinical use. This study offers a hopeful strategy to combat schistosomiasis and reduce its global impact., (© 2024. The Author(s).)

Knee osteoarthritis is a chronic joint disease mainly characterized by cartilage degeneration. The treatment is challenging due to the lack of blood vessels and nerve supplies in cartilaginous tissue, causing a prominent limitation of regenerative capacity. Hence, we investigated the cellular promotional and anti-inflammatory effects of sericin, Bombyx mori-derived protein, on three-dimensional chondrogenic ATDC5 cell models. The results revealed that a high concentration of sericin promoted chondrogenic proliferation and differentiation and enhanced matrix production through the increment of glycosaminoglycans, COL2A1, COL X, and ALP expressions. SOX-9 and COL2A1 gene expressions were notably elevated in sericin treatment. The proteomic analysis demonstrated the upregulation of phosphoglycerate mutase 1 and triosephosphate isomerase, a glycolytic enzyme member, reflecting the proliferative enhancement of sericin. The differentiation capacity of sericin was indicated by the increased expressions of procollagen12a1, collagen10a1, rab1A, periostin, galectin-1, and collagen6a3 proteins. Sericin influenced the differentiation capacity via the TGF-β signaling pathway by upregulating Smad2 and Smad3 while downregulating Smad1, BMP2, and BMP4. Importantly, sericin exhibited an anti-inflammatory effect by reducing IL-1β, TNF-α, and MMP-1 expressions and accelerating COL2A1 production in the early inflammatory stage. In conclusion, sericin demonstrates potential in promoting chondrogenic proliferation and differentiation, enhancing cartilaginous matrix synthesis through glycolysis and TGF-β signaling pathways, and exhibiting anti-inflammatory properties., (© 2024. The Author(s).)

Human gnathostomiasis is a food-borne zoonotic helminthic infection widely reported in Latin America, Asia, and Southeast Asia. Consuming raw, or under-cooked fresh-water fish is the leading cause of this helminthic infection, which is clinically characterized by signs of inflammation, itching sensation, or irritation with migratory swelling. Neurological symptoms resulting from neurognathostomiasis vary, and there is scant information due to the rareness of patient brain samples. This study aimed to demonstrate the first evidence of human neurognathostomiasis by the detection of Gnathostoma spinigerum larva in patient's brain during craniotomy, supported by histopathological, immunological and proteomic evidence. Clinical symptoms were obtained from medical history and physical examination with laboratory investigations, including magnetic resonance imaging (MRI), left temporal craniotomy, histopathology of brain tissue, and Western blot analysis, were performed to elucidate the causative pathogens for diagnosis. In addition, the host-parasite interaction of the parasite invading the patient's brain was characterized through proteomics. Histopathology revealed worms with the characteristic cuticular spines of G. spinigerum which were detected and identified. These histopathological findings were consistent with a positive Western blot showing a 24-kDa reactive-band for gnathostomiasis. Proteomic analysis revealed the presence of G. spinigerum serpin and serine protease in the patient's serum. Moreover, the leucine-rich alpha-2-glycoprotein was indicated as a systemic biomarker of early brain injury related to invasion by G. spinigerum. Therefore, our study provides the initial evidence of human neurognathostomiasis due to G. spinigerum larval invasion along with successful craniotomy and proven larval detection including complete follow-up, and the disease prognosis after surgical treatment., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Inc. on behalf of International Association of Food and Waterborne Parasitology.)

Background Aims: Despite advancements in wound care, wound healing remains a challenge, especially in individuals with type 2 diabetes. Cell sheet technology has emerged as an efficient and promising therapy for tissue regeneration and wound repair. Among these, bilayered human keratinocyte-fibroblast cell sheets constructed using temperature-responsive culture surfaces have been shown to mimic a normal tissue-like structure and secrete essential cytokines and growth factors that regulate the wound healing process., Methods: This study aimed to evaluate the safety and therapeutic potential of human skin cell sheets to treat full-thickness skin defects in a rat model of type 2 diabetes., Results: Our findings demonstrate that diabetic wounds transplanted with bilayered cell sheets resulted in accelerated re-epithelialization, increased angiogenesis, enhanced macrophage polarization and regeneration of tissue that closely resembled healthy skin. In contrast, the control group that did not receive cell sheet transplantation presented characteristic symptoms of impaired and delayed wound healing associated with type 2 diabetes., Conclusions: The secretory cytokines and the upregulation of Nrf2 expression in response to cell sheet transplantation are believed to have played a key role in the improved wound healing observed in diabetic rats. Our study suggests that human keratinocyte-fibroblast cell sheets hold great potential as a therapeutic alternative for diabetic ulcers., Competing Interests: Declaration of Competing Interest The authors declared no competing interests., (Copyright © 2024 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.)

Sericin, a natural protein derived from Bombyx mori, is known to ameliorate liver tissue damage; however, its molecular mechanism remains unclear. Herein, we aimed to identify the possible novel targets of sericin in hepatocytes and related cellular pathways. RNA sequencing analysis indicated that a low dose of sericin resulted in 18 differentially expressed genes (DEGs) being upregulated and 68 DEGs being downregulated, while 61 DEGs were upregulated and 265 DEGs were downregulated in response to a high dose of sericin (FDR ≤ 0.05, fold change > 1.50). Functional analysis revealed that a low dose of sericin regulated pathways associated with the complement and coagulation cascade, metallothionine, and histone demethylate (HDMs), whereas a high dose of sericin was associated with pathways involved in lipid metabolism, mitogen-activated protein kinase (MAPK) signaling and autophagy. The gene network analysis highlighted twelve genes, A2M, SERPINA5, MT2A, MT1G, MT1E, ARID5B, POU2F1, APOB, TRAF6, HSPA8, FGFR1, and OGT, as novel targets of sericin. Network analysis of transcription factor activity revealed that sericin affects NFE2L2, TFAP2C, STAT1, GATA3, CREB1 and CEBPA. Additionally, the protective effects of sericin depended on the counterregulation of APOB, POU2F1, OGT, TRAF6, and HSPA5. These findings suggest that sericin exerts hepatoprotective effects through diverse pathways at different doses, providing novel potential targets for the treatment of liver diseases., (© 2024. The Author(s).)

Soil fungal communities play an important role in regulating biogeochemical transformations, yet soil-related fungal pathogens are emerging threats to humans. Our previous studies have revealed the pathogenic Scedosporium species in soils samples from public parks with high human activities in Thailand. However, measurement and survey of soil fungal communities in other areas with high human/animal activities, such as the pigsty, are poorly determined. In this study, soil fungal pathogens from a pigsty were isolated and identified. Soil samples were collected from the surrounding drainage areas. Fungal species were identified using morphological and molecular analyses. Isolation of soil samples from the pigsty revealed at least 11 species that have been identified. The most abundant fungal species belonged to genera Aspergillus and Penicillium. Moreover, Scedo-Select III culturing and phylogenetic analysis with β-tubulin gene sequencing revealed the three environmental isolates of Scedosporium species, which were consistent with the S.apiospermum. These three Scedosporium isolates were susceptible to voriconazole and caused pathological characteristics of scedosporiosis similar to S. apiospermum in vivo. In conclusion, our findings contribute towards a better understanding of soil-borne pathogenic fungi in the pigsty. The isolation of Scedosporium species with pathogenic potentials in the present study can be beneficial for the management of public health surveillance, epidemiologists, as well as physicians to reduce the risk of soil fungal contamination among pigsty workers.

Candida albicans is an opportunistic fungal infectious agent that can cause cutaneous candidiasis in humans. Biofilms formation of C. albicans is thought to be the major cause of antifungal drug resistance. Despite numerous studies conducted on C. albicans biofilms, a comprehensive understanding of how C. albicans biofilms induced cutaneous candidiasis in humans and the development of a more effective targeted therapy remain poorly investigated. Available animal models of cutaneous candidiasis and in vitro human skin cell cultures do not fully reflect the actual human skin microenvironment or the disease pathogenesis. We investigated the molecular pathology of C. albicans infection using an in vitro biofabricated 3D skin. This in vitro biofabricated 3D skin comprises a fully humanized three-dimensional (3D) skin equivalent, consisting of a stratified terminally differentiated epidermis and an underlying dermal compartment. Antifungal drug susceptibility testing, histological and electron microscopy study, biofilms study, and pro-inflammatory cytokines analysis were conducted in C. albicans infected skin. Histological results revealed that C. albicans covered and produced biofilm on the in vitro biofabricated 3D skin, invading the skin compartments including epidermis and dermis. Elevation of proinflammatory cytokines including MMP-9, IL-1β, TNF-α, and IL-5 were examined in the C. albicans infected skin. However, treatment with itraconazole reduced the pathology of C. albicans infection. This study provides an alternative pathological model of cutaneous candidiasis, which can physiologically represent a close-up event during C. albicans. Moreover, it is rapid, cost-effective, and reproducible of the in vitro biofabricated 3D skin model, and may further highlight the importance of utilizing in vivo-like conditions to improve high-throughput screening for drug discovery against several antifungal drug resistant pathogens.

C. albicans is one of the most important species of fungi known to produce biofilms on installed medical devices. The environment surrounding the fungi influences the development of the biofilm. Temperature is known to affect the yeast-to-hypha transition of C. albicans, but the impact of this factor on biofilm formation is still not understood. This study aimed to investigate the effects of temperature (42°C versus 37°C) on the formation of C. albicans biofilms. Three reference C. albicans strains were used: SC 5314, ATCC 90028, and ATCC 96901. Biofilm development was monitored in a series of time intervals, 2, 4, 6, 8, 24, and 48 h, at both 37°C and 42°C. Biofilm formation under each condition was evaluated by scanning electron microscopy, crystal violet staining, and 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-(phenylamino)-carbonyl-2H-tetrazoliumhydroxide reduction assay. Our results demonstrated that at 42°C, tested strains of C. albicans could produce a biofilm, but the mass, thickness, and metabolic activity were lower than those of the biofilm formed at 37°C.

Silk sericin-releasing (sericin/polyvinyl alcohol (PVA)/glycerin) scaffolds have been designed for wound dressing applications using different fabrication techniques that influence scaffold antigenicity. The immunological tolerance of scaffolds depends on the balance of immunogenic and tolerogenic responses modulated by dendritic cells (DCs). An in vivo skin implantation model was used to compare the tolerogenic effect of sericin/PVA/glycerin scaffolds prepared by freeze-drying versus salt-leaching techniques, using an Allevyn® scaffold as a control. Immunohistochemical and histopathological studies were performed to evaluate tolerogenic DCs (CD206+), immunogenic DCs (CD83+), regulatory T-cells (FOXP3+ and CTLA-4), a proinflammatory cytokine (interleukin 33: IL-33), a stress marker (heat shock protein 60; HSP60), histopathological changes and related inflammatory cells. It was found that both sericin/PVA/glycerin scaffolds were tolerogenic and induced early activated Treg functions, while the Allevyn® scaffold was immunogenic. However, the tolerance of the freeze-dried sericin/PVA/glycerin scaffolds was not as consistent as the salt-leached sericin/PVA/glycerin scaffolds, indicated by the low level of CTLA-4 expression. This was probably due to molecular cross-linking and the morphological and mechanical properties of the freeze-drying technique, which would enhance the immune response. Severe inflammatory responses (including mast cell degranulation and foreign body giant cell accumulation) and histopathological changes (including fat infiltration and fibrosis formation) were mainly found with the Allevyn® scaffold, presumably from its architecture and chemical composition, especially polyurethane. The up-regulation of IL-33 and HSP60 with the Allevyn® scaffold was correlated with the inflammatory and pathological levels. Our findings suggested that salt-leached sericin/PVA/glycerin scaffolds were tolerogenic, induced a low inflammatory response and were appropriate for wound dressing applications.

The pathogenesis of congenital hydronephrosis in laboratory animals has been studied for many years, yet little is known about the underlying mechanism of this disease. In this study, we investigated a MS-based comparative proteomics approach to characterize the differently expressed proteins between kidney tissue samples of ICR/Mlac-hydro and wild-type mice. Interestingly, proteomic results exhibited several mitochondrial protein alterations especially the up-regulation of 60 kDa heat shock protein (Hsp60), stress-70 protein (GRP75) dysfunction, and down-regulation of voltage-dependent anion-selective channel protein 1 (VDAC-1). The results demonstrated that mitochondrial alteration may lead to inadequate energy-supply to maintain normal water reabsorption from the renal tubule, causing hydronephrosis. Moreover, the alteration of cytoskeleton proteins in the renal tubule, in particular the up-regulation of tubulin beta-4B chain (Tb4B) and N-myc downstream-regulated gene 1 protein (Ndr-1) may also be related due to their fundamental roles in maintaining cell morphology and tissue stability. In addition, cytoskeletal alterations may consequence to the reduction of glyceraldehydes-3-phosphate dehydrogenase (GAPDH), cytoplasmic enzyme, which modulates the capacity of structural proteins. Our findings highlight a number of target proteins that may play a crucial role in congenital hydronephrosis and emphasize that the disorder of mitochondria and cytoskeleton proteins may be involved.

The mechanisms leading to cerebral malaria (CM) are not completely understood. Brain edema has been suggested as having an important role in experimental CM. In this study, CBA/CaH mice were infected with Plasmodium berghei ANKA blood-stage and when typical symptoms of CM developed on day 7, brain tissues were processed for electron-microscopic and immunohistochemical studies. The study demonstrated ultrastructural hallmarks of cerebral edema by perivascular edema and astroglial dilatation confirming existing evidence of vasogenic and cytogenic edema. This correlates closely with the clinical features of CM. An adaptive response of astrocytic activity, represented by increasing glial fibrillary acidic protein (GFAP) expression in the perivascular area and increasing numbers of large astrocyte clusters were predominately found in the CM mice. The presence of multivesicular and lamellar bodies indicates the severity of cerebral damage in experimental CM. Congestion of the microvessels with occluded white blood cells (WBCs), parasitized red blood cells (PRBCs) and platelets is also a crucial covariate role for CM pathogenesis.

Chronic renal impairment can lead to bone deterioration and abnormal bone morphology, but whether hydronephrosis is associated with bone loss remains unclear. Herein, we aimed to use computer-assisted bone histomorphometric technique to investigate microstructural bone changes in Imprinting Control Region (ICR) mice with a spontaneous mutation that was associated with bilateral nonobstructive hydronephrosis (ICR/Mlac-hydro). The results showed that 8-week-old ICR/Mlac-hydro mice manifested decreases in trabecular bone number and thickness, and an increased trabecular separation, thereby leading to a reduction in trabecular bone volume compared with the wild-type mice. Furthermore, histomorphometric parameters related to both bone resorption and formation, that is, eroded surface, osteoclast surface, and osteoblast surface, were much lower in ICR/Mlac-hydro mice than in the wild type. A decrease in moment of inertia was found in ICR/Mlac-hydro mice, indicating a decrease in bone strength. In conclusion, ICR/Mlac-hydro mice exhibited trabecular bone loss, presumably caused by marked decreases in both osteoblast and osteoclast activities, which together reflected abnormally low bone turnover. Thus, this mouse strain appeared to be a valuable model for studying the hydronephrosis-associated bone disease., (Copyright © 2013 Wiley Periodicals, Inc.)

To understand the relationship among cholesterolemia, hyperglycemic stage in non obese type 2 diabetes mellitus, and histological perturbations on liver, retina, hippocampus, and Harderian gland, we maintained rat on a diet high in cholesterol for fourteen weeks, then analyzed blood lipid profiles, blood glucose, hepatic enzymes, and microscopic lesion of those tissues. We observed that high cholesterol-treated rat elevated in cholesterol and low density lipoprotein with not correlated to hyperglycemia. Histopathological changing in Goto-Kakizaki rat on liver (microvesicular steatosis) and Harderain gland (tubular lesions) were related to hyperglycemic effect rather than cholesterolemic effect. These may be related to hypoinsulinemic characteristic of this diabetic model. However increasing pyknotic nuclei on hippocampus and reducing of retinal ganglionic cell were related to the high level of cholesterol loaded with synergized effect due to diabetic stage.

The pathogenic mechanisms underlying the occurrence of cerebral malaria (CM) are still incompletely understood but, clearly, cerebral complications may result from concomitant microvessel obstruction and inflammation. The extent to which brain edema contributes to pathology has not been investigated. Using the model of P. berghei ANKA infection, we compared brain microvessel morphology of CM-susceptible and CM-resistant mice. By quantitative planimetry, we provide evidence that CM is characterized by enlarged perivascular spaces (PVS). We show a dramatic aquaporin 4 (AQP4) upregulation, selectively at the level of astrocytic foot processes, in both CM and non-CM disease, but significantly more pronounced in mice with malarial-induced neurological syndrome. This suggests that a threshold of AQP4 expression is needed to lead to neurovascular pathology, a view that is supported by significantly higher levels in mice with clinically overt CM. Numbers of intravascular leukocytes significantly correlated with both PVS enlargement and AQP4 overexpression. Thus, brain edema could be a contributing factor in CM pathogenesis and AQP4, specifically in its astrocytic location, a key molecule in this mechanism. Since experimental CM is associated with substantial brain edema, it models paediatric CM better than the adult syndrome and it is tempting to evaluate AQP4 in the former context. If AQP4 changes are confirmed in human CM, it may represent a novel target for therapeutic intervention.