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Search Results (274)

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10 pages, 607 KiB  
Article
Terbinafine Resistance in Trichophyton Strains Isolated from Humans and Animals: A Retrospective Cohort Study in Italy, 2016 to May 2024
by Silvia Crotti, Deborah Cruciani, Michela Sabbatucci, Sara Spina, Vincenzo Piscioneri, Martina Torricelli, Roberta Calcaterra, Claudio Farina, Luigi Pisano and Manuela Papini
J. Clin. Med. 2024, 13(18), 5493; https://doi.org/10.3390/jcm13185493 (registering DOI) - 17 Sep 2024
Abstract
Background: In recent decades, globalization and international migration have increased the spread of infectious agents, including dermatophytes. Although considered minor infections, dermatophytoses are highly contagious, and they significantly reduce the quality of life, inducing itching, burning, sleep disturbances, and even depressive states. Moreover, [...] Read more.
Background: In recent decades, globalization and international migration have increased the spread of infectious agents, including dermatophytes. Although considered minor infections, dermatophytoses are highly contagious, and they significantly reduce the quality of life, inducing itching, burning, sleep disturbances, and even depressive states. Moreover, the increasing resistance to antifungals threats the public health and burdens the costs for the healthcare system. Methods: DermaGenius® Resistance Multiplex real-time PCR assay allowed to analyze the terbinafine susceptibility/resistance of 172 Trichophyton strains, which were isolated from human and animal samples collected from 2016 to May 2024 and previously identified by Sanger sequencing. Results: All the 11 animal strains belonged to the T. interdigitale/T. mentagrophytes complex and tested terbinafine sensitive. Out of 161 human strains, 9 (5.6%) showed terbinafine resistance and 7 (4.3%) were identified as T. indotineae. Conclusions: This study provides preliminary data about behavior toward antifungals in animals and finalizes the scientific information currently available about human strains, highlighting the importance of the One Health concept. Moreover, it supports the relevant role of T. indotineae as an emerging dermatophyte with high proportion of terbinafine resistance. Full article
(This article belongs to the Section Dermatology)
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<p>Dermatophyte species obtained by Sanger sequencing and their genotypic terbinafine sensitivity/resistance (S/R) analyzed by DermaGenius<sup>®</sup> Resistance Multiplex real-time PCR assay in human samples (HS) and animal samples (AS).</p>
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13 pages, 268 KiB  
Review
The Zoonotic Potential of Fungal Pathogens: Another Dimension of the One Health Approach
by Reetu Kundu, Yashik Bansal and Nidhi Singla
Diagnostics 2024, 14(18), 2050; https://doi.org/10.3390/diagnostics14182050 - 15 Sep 2024
Viewed by 284
Abstract
Zoonotic diseases are caused by viruses, bacteria, fungi and parasites and they comprise about 75% of all emerging infectious diseases. These can be transmitted via the direct (scratches on skin or animal bites) or indirect mode (through environmental shedding of infectious agent by [...] Read more.
Zoonotic diseases are caused by viruses, bacteria, fungi and parasites and they comprise about 75% of all emerging infectious diseases. These can be transmitted via the direct (scratches on skin or animal bites) or indirect mode (through environmental shedding of infectious agent by the infected animal) of transmission. Environmental changes, whether in the form of urbanization, industrialization or destruction of wildlife habitats, lead to more human invasion in wildlife areas, subsequently leading to an increased passage of animals towards human dwellings and more exposure to animals, making humans susceptible to these infections. Climate change is another major factor. Global warming and the evolving thermotolerance of fungi, adapting more to human body temperature than their saprophytic nature, is leading to the emergence of humans as new hosts for fungi. The domestication of animals, rising populations, enhanced tourism, migratory populations, intrusions into wildlife, etc., are other known factors. Zoonotic fungal infections have long been neglected and are now gaining due attention. In this review, we briefly discuss the various aspects currently known for zoonotic fungal infections and bring forth the importance of this particular issue to be addressed in a timely manner. Full article
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
13 pages, 2888 KiB  
Article
Epidemiology of Onychomycosis in the United States Characterized Using Molecular Methods, 2015–2024
by Aditya K. Gupta, Tong Wang, Shruthi Polla Ravi, Avantika Mann, Sara A. Lincoln, Hui-Chen Foreman and Wayne L. Bakotic
J. Fungi 2024, 10(9), 633; https://doi.org/10.3390/jof10090633 - 5 Sep 2024
Viewed by 349
Abstract
Onychomycosis is a recalcitrant fungal infection of the nail unit that can lead to secondary infections and foot complications. Accurate pathogen identification by confirmatory testing is recommended to improve treatment outcomes. In this study, we reviewed the records of 710,541 patients whose nail [...] Read more.
Onychomycosis is a recalcitrant fungal infection of the nail unit that can lead to secondary infections and foot complications. Accurate pathogen identification by confirmatory testing is recommended to improve treatment outcomes. In this study, we reviewed the records of 710,541 patients whose nail specimens were sent to a single molecular diagnostic laboratory between 2015 and 2024. PCR testing revealed a more comprehensive spectrum of pathogens than previously reported, which was corroborated by the demonstration of fungal invasion on histopathology. Consistent with our current understanding, the T. rubrum complex (54.3%) are among the most common pathogens; however, a significant portion of mycology-confirmed diagnoses were caused by the T. mentagrophytes complex (6.5%), Aspergillus (7.0%) and Fusarium (4.5%). Females were significantly more likely to be infected with non-dermatophytes molds (NDMs; OR: 2.0), including Aspergillus (OR: 3.3) and Fusarium (OR: 2.0), and yeasts (OR: 1.5), including Candida albicans (OR: 2.0) and C. parapsilosis (OR 1.6), than males. The T. mentagrophytes complex became more prevalent with age, and conversely the T. rubrum complex became less prevalent with age. Patients aged ≥65 years also demonstrated a higher likelihood of contracting onychomycosis caused by NDMs (OR: 1.6), including Aspergillus (OR: 2.2), Acremonium (OR: 3.5), Scopulariopsis (OR: 2.9), Neoscytalidium (OR: 3.8), and yeasts (OR: 1.8), including C. albicans (OR: 1.9) and C. parapsilosis (OR: 1.7), than young adults. NDMs (e.g., Aspergillus and Fusarium) and yeasts were, overall, more likely to cause superficial onychomycosis and less likely to cause dystrophic onychomycosis than dermatophytes. With regards to subungual onychomycosis, Aspergillus, Scopulariopsis and Neoscytalidium had a similar likelihood as dermatophytes. The advent of molecular diagnostics enabling a timely and accurate pathogen identification can better inform healthcare providers of appropriate treatment selections and develop evidence-based recommendations. Full article
(This article belongs to the Special Issue Personalized Mycology)
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<p>PAS-stained nail specimens: (<b>A</b>,<b>D</b>) a subungual infection pattern evidenced by the presence of fungal elements in the subungual keratin; (<b>B</b>,<b>E</b>) a superficial infection pattern evidenced by the presence of fungal elements within the dorsal aspect of the nail plate; and (<b>C</b>,<b>F</b>) a dystrophic infection pattern evidenced by the presence of fungal elements in the nail plate and in the subungual keratin. Fungal elements are indicated by black arrows.</p>
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<p>Distribution of fungal agents in patients with mycology-confirmed onychomycosis diagnoses by PCR and histopathology. Results are stratified by year and presented as mean ± SD; mixed detections and un-speciated samples are not shown.</p>
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<p>Likelihood of: (<b>A</b>) NDMs; and (<b>B</b>) yeasts in causing subungual onychomycosis, with moderate-to-florid fungal element quantities, compared to dermatophytes. Data points represent OR (95% CI), grey-shaded bars represent the prevalence of the respective NDM/yeast target, the reference (dotted) line represents dermatophytes with an OR of 1.</p>
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<p>Likelihood of: (<b>A</b>) NDMs; and (<b>B</b>) yeasts in causing superficial onychomycosis compared to dermatophytes. Data points represent OR (95% CI), grey-shaded bars represent the prevalence of the respective NDM/yeast target, the reference (dotted) line represents dermatophytes with an OR of 1.</p>
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<p>Likelihood of: (<b>A</b>) NDMs; and (<b>B</b>) yeasts in causing dystrophic onychomycosis compared to dermatophytes. Data points represent OR (95% CI), grey-shaded bars represent the prevalence of the respective NDM/yeast target, the reference (dotted) line represents dermatophytes with an OR of 1.</p>
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12 pages, 2117 KiB  
Article
Extended Use of Topical Efinaconazole Remains Safe and Can Provide Continuing Benefits for Dermatophyte Toenail Onychomycosis
by Aditya K. Gupta and Elizabeth A. Cooper
J. Fungi 2024, 10(9), 620; https://doi.org/10.3390/jof10090620 - 30 Aug 2024
Viewed by 355
Abstract
Introduction: Efinaconazole 10% topical solution labeling for onychomycosis describes phase III trials of 12 months of treatment; the slow growth of onychomycotic nails suggests a longer treatment period may increase efficacy. We present here the first evaluation of extended use of efinaconazole 10% [...] Read more.
Introduction: Efinaconazole 10% topical solution labeling for onychomycosis describes phase III trials of 12 months of treatment; the slow growth of onychomycotic nails suggests a longer treatment period may increase efficacy. We present here the first evaluation of extended use of efinaconazole 10% topical solution for up to 24 months. Materials and Methods: Enrolled patients (n = 101) had one target great toenail with mild to moderate distal lateral subungual onychomycosis and applied efinaconazole 10% topical solution to all affected toenails once daily for 18 months (EFN18) or 24 months (EFN24). Efficacy and safety were evaluated at each visit by visual review and mycology sampling. Results: Regarding the target toenail for patients treated for 24 months (EFN24), mycological cure (negative microscopy and culture) was 66.0% at Month 12, increasing to 71.7% at Month 24; effective cure (mycological cure and ≤10% affected nail) was 13.2% at Month 12, rising to 22.6% at Month 24. Mild to moderate application site reactions (symptoms of erythema/scaling) were the only efinaconazole-related reactions, in eight patients (7.9%). No systemic efinaconazole events or drug interactions were found. Patients aged 70 years or more had similar efficacy to younger patients at all time periods and did not show any increased treatment risks. Thinner nails exhibited better clearance versus thicker nails. A higher proportion of patients with Trichophyton mentagrophytes complex infection experienced application site reactions (35.7%), and a higher effective cure was found at Month 24 versus T. rubrum patients. Conclusion: There is a trend of increasing mycological cure and effective cure beyond Month 12 to Month 24, without an increased safety risk. The enrolled population in this trial was significantly older than in the phase III trials, with a greater degree of onychomycosis severity; however, increased age did not appear to reduce the chance of efficacy to Month 24 in this study. Our data suggest that lack of ability to clear nail dystrophy remains a significant problem for patients, rather than any lack of efinaconazole action over long-term treatment periods. Full article
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<p>Mycological and effective cure rates by treatment group.</p>
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<p>Patient photos (EFN24) demonstrating onychomycosis cure at Month 24.</p>
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<p>Efinaconazole for 24 months (EFN24 ONLY): cure rates by age.</p>
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<p>Efinaconazole for 24 months (EFN24 ONLY): cure by nail thickness.</p>
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<p>Efinaconazole for 24 months (EFN24 ONLY): cure rates by organism.</p>
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11 pages, 1003 KiB  
Article
Physicochemical Characterization of Carao Honey Flour (Cassia grandis) and Its Effects on the Sensory Attributes in a Cookie
by Jhunior Marcía Fuentes, Manuel de Jesús Álvarez Gil, Héctor Zumbado Fernández, Ismael Montero-Fernández, Daniel Martín-Vertedor, Ajitesh Yadav and Ricardo S. Aleman
Appl. Sci. 2024, 14(17), 7502; https://doi.org/10.3390/app14177502 - 25 Aug 2024
Viewed by 582
Abstract
Carao honey is a potential functional ingredient that can generate added value to food products enriched by reducing waste. In many countries, carao has been utilized for therapeutic purposes since this type of plant extract can inhibit the growth against the most common [...] Read more.
Carao honey is a potential functional ingredient that can generate added value to food products enriched by reducing waste. In many countries, carao has been utilized for therapeutic purposes since this type of plant extract can inhibit the growth against the most common dermatophytes, a pathogenic fungus that grows on the skin, mucous membranes, hair, nails, feathers, and other body surfaces, causing ringworm and related diseases. The physicochemical characteristics of the cookies were also investigated, which included aW, proximate analysis, hardness, and color. A sensory study was carried out to determine the rejection threshold, acceptability, purchase intent, and general taste of 90 consumers. The gluten-free cookies were prepared with carao honey as a partial substitute for rice flour in different percentages (0%, 2.5%, 5%, and 10%). The carao honey proximate composition, rheological properties, and pasting properties were analyzed. It was emphasized that incorporating carao honey into cookies improves the ingredient’s sustainability. The impact of carao on the physicochemical and sensory properties of cookies was evident in color, flavor, and smell. According to the results, only samples with 2.5% carao were accepted regarding flavor and smell. Overall, liking scores, age, and flavor were the most significant predictors of purchase intent. The information given to panelists did not significantly (p < 0.05) influence liking scores only for cookies. According to physicochemical analysis, carao honey flour was high in carbohydrates (88%). The incorporation of carao produced cookies with darker colors and a softer texture. The study demonstrated that carao flour could be included to produce sensorial accepted cookies at a 2.5% addition level as an alternative in the food industry that benefits from environmental sustainability and, at the nutritional level, improves the availability of nutrients, mainly sugars and proteins. Full article
(This article belongs to the Special Issue Topical Advisory Panel Members’ Collection Series: Functional Foods)
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<p>Flowchart of carao honey process.</p>
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<p>Viscosity profiles of rice flours determined by RVA. The 0%, 2.5%, 5%, and 10% C-CHF treatments correspond to the CHF addition of 0%, 2.5%, 5%, and 10%, respectively.</p>
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<p>Heat flux of rice flours determined by DSC. The 0%, 2.5%, 5%, and 10% C-CHF treatments correspond to the CHF addition of 0%, 2.5%, 5%, and 10%, respectively.</p>
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17 pages, 3550 KiB  
Article
Dermatophytosis in Companion Animals in Portugal: A Comprehensive Epidemiological Retrospective Study of 12 Years (2012–2023)
by Ricardo Lopes, Andreia Garcês, Augusto Silva, Paula Brilhante-Simões, Ângela Martins, Luís Cardoso, Elsa Leclerc Duarte and Ana Cláudia Coelho
Microorganisms 2024, 12(8), 1727; https://doi.org/10.3390/microorganisms12081727 - 22 Aug 2024
Viewed by 511
Abstract
Dermatophytosis, commonly referred to as ringworm, is a common superficial fungal infection in companion animals and humans. Between 2012 and 2023, plucked hair and scraped scale samples from domestic dogs and cats with clinical suspicion of dermatophytosis were collected from 355 veterinary medical [...] Read more.
Dermatophytosis, commonly referred to as ringworm, is a common superficial fungal infection in companion animals and humans. Between 2012 and 2023, plucked hair and scraped scale samples from domestic dogs and cats with clinical suspicion of dermatophytosis were collected from 355 veterinary medical centres across mainland Portugal. A total of 4716 animal samples were inoculated onto DERM agar, incubated at 25 °C for up to 4 weeks, and periodically examined macro- and micro-scopically to observe and evaluate fungal growth. Of these, 271 samples were removed due to contaminant fungi. Of the 568 positive cultures, the highest number were from the North (48.1%; 95% CI: 44.0–52.2%) and Centre (32.4%; 95% CI: 28.7–36.4%) regions. Microsporum canis was the most frequently isolated species (63.9%), followed by Trichophyton spp. (20.3%) and Nannizia gypsea (formerly Microsporum gypseum) (8.1%). Felines exhibited a higher frequency (17.4%) compared with dogs (9.1%) (p < 0.001). In dogs, the Yorkshire Terrier, West Highland White Terrier, Miniature Pinscher, Dalmatian and Miniature Schnauzer demonstrated a significant predisposition to dermatophytosis (p < 0.05). In cats, the Persian and Scottish Fold breeds were significantly predisposed (p < 0.05). No significant differences were found between sexes (p > 0.05). These findings underscore dermatophytosis as an increasing public health concern due to its zoonotic and contagious nature, providing comprehensive insights into the epidemiology of dermatophytosis in Portugal. Full article
(This article belongs to the Section Veterinary Microbiology)
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<p>Dermatophytosis (ectothrix) in a cat. Arthrospores on the exterior of the hair shaft (Lactophenol Cotton Blue stain, 400×).</p>
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<p>Samples were inoculated onto DERM agar plates (bioMérieux, Marcy–l’Étoile, France), incubated at 25 °C. <span class="html-italic">Trichophyton</span> spp. with colonies smooth or powdery, with colours ranging from white to cream or tan (<b>A</b>). The reverse side of these colonies can show the colour red (<b>B</b>) due to the phenol red in the agar as a pH indicator when dermatophyte positive.</p>
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<p><span class="html-italic">Trichophyton</span> spp. typically have numerous microconidia forming dense clusters. These microconidia are hyaline, smooth-walled and predominantly spherical or subspherical. Some species also exhibit cigar-shaped macroconidia with smooth, thin walls. Additional microscopic features can include spiral hyphae and chlamydospores, varying among different species (Lactophenol Cotton Blue stain, 1000×).</p>
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<p><span class="html-italic">Microsporum canis</span> presents septate hyphae, spindle-shaped macroconidia with 6–15 compartments and asymmetrical ends and microconidia. The macroconidia are long, rough and have dense outer walls (Lactophenol Cotton Blue stain, 1000×).</p>
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<p><span class="html-italic">Nannizzia gypsea</span> (formerly <span class="html-italic">Microsporum gypseum</span>) shows septate hyphae and a significant number of macroconidia and microconidia. The macroconidia are fusiform and symmetrical with rounded ends, containing 3–6 compartments. Microconidia are moderately numerous and are located along the hyphae (Lactophenol Cotton Blue stain, 1000×).</p>
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<p><span class="html-italic">Nannizzia nana</span> (formerly <span class="html-italic">Microsporum nanum</span>) exhibits septate hyphae and pear-shaped macroconidia, usually with two compartments. Microconidia are less common and smaller compared to those in other species (Lactophenol Cotton Blue stain, 1000×).</p>
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<p>Spatial distribution, according to the distribution of animals from the different districts of Portugal, of the 4445 animals included in this study (map drawn in paintmaps.com; accessed on 23 June 2024).</p>
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<p>Map of continental Portugal showing a categorical representation of average frequencies of dermatophyte diagnosis over 12 years (2012–2023) per district and region (NUTS2) (map drawn in mapinseconds.com; accessed on 23 June 2024).</p>
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14 pages, 3922 KiB  
Article
Microsporum canis Causes Cutaneous and Extracutaneous Feline Dermatophytic Pseudomycetomas: Molecular Identification and Clinicopathological Characteristics
by Stefan Hobi, Wing Yan Jacqueline Tam, May Tse, Omid Nekouei, Yingfei Chai, Fraser I. Hill, Edmund Cheung, Wietz Botes, Francois Saulnier-Troff, Colin T. McDermott and Vanessa R. Barrs
J. Fungi 2024, 10(8), 576; https://doi.org/10.3390/jof10080576 - 14 Aug 2024
Viewed by 743
Abstract
Dermatophytic pseudomycetoma (DPM) is a rarely reported invasive fungal infection of humans and animals, especially cats. This study aimed to identify dermatophytes, breed associations, and the frequency of extracutaneous (EC) involvement in feline DPM. Electronic records and formalin-fixed paraffin-embedded tissue (FFPET) from 32 [...] Read more.
Dermatophytic pseudomycetoma (DPM) is a rarely reported invasive fungal infection of humans and animals, especially cats. This study aimed to identify dermatophytes, breed associations, and the frequency of extracutaneous (EC) involvement in feline DPM. Electronic records and formalin-fixed paraffin-embedded tissue (FFPET) from 32 suspected DPM cases in 30 cats were retrieved from a diagnostic laboratory between 2018 and 2024. To confirm DPM and molecular identity, DNA was extracted from FFPET for ITS2 sequencing, and immunohistochemistry was performed on PCR-negative cases. All cases were confirmed as DPM. Microsporum canis was the only dermatophyte identified. The sensitivity and specificity of ITS2 sequencing for M. canis identification in FFPET were 22/32 (68.8%) and 21/22 (95.5%), respectively. Exotic (36.7%) and Persian (23.3%) but not British breeds (26.3%) were over-represented compared to feline admissions at an affiliated veterinary hospital (8.5%, p < 0.001; 3%, p < 0.001; 21.6%, p = 0.817, respectively). Five cases (16.7%) had EC lesions; two had intra-abdominal masses; two had oral cavity masses, including one which extended into the cranial vault; and one had superficial cervical lymph node invasion. Exotic and Persian breeds are over-represented for DPM and M. canis is the primary cause. EC lesions of DPM may occur more commonly than previously thought. Full article
(This article belongs to the Special Issue Advances in Human and Zoonotic Dermatophytoses)
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<p>Case 3 (<a href="#jof-10-00576-t001" class="html-table">Table 1</a>) was presented with diffuse ulceration and thickening of ventral abdominal skin (<b>A</b>). After 3-month treatment with itraconazole then terbinafine, the remaining affected skin (<b>B</b>) was resected en bloc, and an enlarged left superficial cervical lymph node (<b>C</b>) was biopsied. The cat developed a recurrent focal dermatophytic pseudomycetoma the following year (<b>D</b>) involving a hind-limb digit.</p>
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<p>Diagnostic imaging findings in a cat with an intra-abdominal dermatophytic pseudomycetoma and presentation for constipation (Case 8, <a href="#jof-10-00576-t001" class="html-table">Table 1</a>). Right lateral radiograph (<b>A</b>) and sagittal (<b>B</b>), transverse (<b>C</b>), and coronal (<b>D</b>) computed tomographic images show a soft-tissue density retroperitoneal sub-lumbar mass dorsal to the colon (yellow arrowheads), extending from the cranial border of L6 cranially through the pelvic inlet caudally where it obliterates the rectal lumen. Proximal to the rectum, the colon is distended with faeces.</p>
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<p>A 10-year-old Exotic shorthair (Case 10, <a href="#jof-10-00576-t001" class="html-table">Table 1</a>) with a left-head tilt, stertorous respiration, left retromolar oral mass (<b>A</b>,<b>B</b>), a large mass over the left submandibular/auricular region (asterisk (<b>A</b>,<b>C</b>,<b>D</b>)), and multiple nodular masses on the dorsal head and neck. A CT scan of the head showed the left-sided facial mass was multilobulated and extended across tissue planes into the oral cavity at the level of the temporomandibular joint, obliterating the caudal nasopharynx (<b>D</b>). The mass extended intracranially through a markedly widened oval foramen at the level of the TMJ, showed heterogeneous contrast uptake, and displaced the cerebrum dorsally ((<b>C</b>), arrows).</p>
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<p>Histological and staining characteristics of a dermatophytic pseudomycetoma in the superficial cervical lymph node in Case 3, <a href="#jof-10-00576-t001" class="html-table">Table 1</a>. The lymph node is effaced by a pyogranulomatous cellulitis with multifocal amorphous foci and occasional lymphoid aggregate, hematoxylin and eosin (<b>A</b>) (H&amp;E), 100×; (<b>B</b>) higher magnification of these amorphous foci reveals matts of fungal hyphae surrounded by epithelioid macrophages (arrow), fewer multinucleated giant cells (asterisks), and neutrophils (arrowhead), H&amp;E, 400×, scale bar = 100 µm.; (<b>C</b>) fungal hyphae with bulbous wall highlighted by Periodic Acid Schiff (PAS), 200×; and (<b>D</b>) Grocott’s methenamine sliver (GMS), 200×; scale bar = 100 µm.</p>
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<p>Immunohistochemical staining of a cutaneous dermatophytic pseudomycetoma, Case 1, <a href="#jof-10-00576-t001" class="html-table">Table 1</a>, stained with an anti-dermatophyte mouse monoclonal antibody (Xceltis GmbH, Mannheim, Germany). Fungal hyphae in the centre of the lesions exhibit positive (brown) staining (<b>A</b>). There is no positive staining of the negative control tissue (<b>B</b>). Scale bar = 50 µm.</p>
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17 pages, 881 KiB  
Review
Is Raman Spectroscopy of Fingernails a Promising Tool for Diagnosing Systemic and Dermatological Diseases in Adult and Pediatric Populations?
by Teresa Tabasz, Natalia Szymańska, Katarzyna Bąk-Drabik, Aleksandra Damasiewicz-Bodzek and Agnieszka Nowak
Medicina 2024, 60(8), 1283; https://doi.org/10.3390/medicina60081283 - 9 Aug 2024
Viewed by 615
Abstract
Background: Raman spectroscopy is a well-known tool used in criminology, molecular biology, and histology. It is also applied to diagnose bone mineral disorders by taking advantage of the similarity of the structure of keratin and bone collagen. Raman spectroscopy can also be [...] Read more.
Background: Raman spectroscopy is a well-known tool used in criminology, molecular biology, and histology. It is also applied to diagnose bone mineral disorders by taking advantage of the similarity of the structure of keratin and bone collagen. Raman spectroscopy can also be used in dermatology and diabetology. The purpose of the present review is to critically evaluate the available research about the use of Raman spectroscopy in the mentioned areas of medicine. Methodology: PubMed was searched for peer-reviewed articles on the subject of use of Raman spectroscopy in bone mineral disorders, dermatology, and diabetes mellitus. Results: Nail keratin and bone collagen are related structural proteins that require disulfide bond for structural stability. Therefore, Raman spectroscopy of keratin may have potential as a diagnostic tool for screening bone quality and distinguishing patients at risk of fracture for reasons different from low bone mineral density (BMD) in the adult women population. Raman spectroscopy can also investigate the changes in keratin’s structure in nails affected by onychomycosis and distinguish between healthy and onychomycosis nail samples. It could also reduce the need for nail biopsy by distinguishing between dermatophytic and non-dermatophytic agents of onychomycosis. Additionally, Raman spectroscopy could expedite the diagnostic process in psoriasis (by assessing the secondary structure of keratin) and in diabetes mellitus (by examining the protein glycation level). Conclusions: In adult populations, Raman spectroscopy is a promising and safe method for assessing the structure of fingernails. However, data are scarce in the pediatric population; therefore, more studies are required in children. Full article
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<p>The principle of operation of Raman spectroscopy.</p>
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<p>The concept behind the use of Raman spectroscopy in the diagnosis of bone structure disorders.</p>
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12 pages, 1586 KiB  
Article
Zoonotic Enteric Nematodes and Dermatophytes in Cat Cafés: An Investigation in the Bangkok Metropolitan Area, Thailand
by Phakjira Sanguansook, Siwaporn Tuangpermsub, Boonyakorn Leelakarnsakul, Sutida Phaisansomsuk, Vachira Hunprasit, Laura Del Río, Waree Niyomtham, Nuvee Prapasarakul and Woraporn Sukhumavasi
Vet. Sci. 2024, 11(8), 358; https://doi.org/10.3390/vetsci11080358 - 7 Aug 2024
Viewed by 1304
Abstract
Cat cafés have gained significant popularity worldwide, offering a unique interface between humans and cats. The present study aims to assess the prevalence of potentially zoonotic endoparasites and dermatophytes from cats living in cat cafés situated in the Bangkok metropolitan area in 2017–2018. [...] Read more.
Cat cafés have gained significant popularity worldwide, offering a unique interface between humans and cats. The present study aims to assess the prevalence of potentially zoonotic endoparasites and dermatophytes from cats living in cat cafés situated in the Bangkok metropolitan area in 2017–2018. Cat fecal samples were subjected to microscopic examination employing centrifugal flotation and centrifugal sedimentation techniques. The hair samples from every cat were cultured on a dermatophyte test medium and Sabouraud dextrose agar and subsequently confirmed by visualization of the typical colony and macroconidia morphology. Findings from 11 cat cafés indicated an 18.2% (2/11) prevalence of gastrointestinal parasites, including Toxocara spp., Ancylostoma spp., Physaloptera spp., and Eucoleus aerophilus. Dermatophytes were prevalent in 16.2% (32/198) of the total number of cats tested, with Microsporum canis being the sole species identified. Notably, the presence of dermatophyte was significantly correlated with the presence of skin lesions and the cats’ origin. In summary, the findings of this study have provided evidence of potentially zoonotic endoparasites and dermatophytes in cats residing in cat cafés. Therefore, it is imperative to heighten awareness and encourage preventive measures among cat café owners and customers to halt the dissemination of these pathogens. Full article
(This article belongs to the Section Veterinary Microbiology, Parasitology and Immunology)
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<p>Geographical distribution of the selected 11 cat cafés for this study located in Bangkok metropolitan area, Thailand, 2017–2018.</p>
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<p>Microscopic findings of endoparasite eggs isolated from cat feces of positive cat café taken at 400× magnification: (<b>A</b>) <span class="html-italic">Toxocara</span> spp. (<b>B</b>) <span class="html-italic">Ancylostoma</span> spp. (<b>C</b>) <span class="html-italic">Physaloptera</span> spp. (<b>D</b>) <span class="html-italic">Eucoleus aerophilus</span>.</p>
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<p><span class="html-italic">Demodex gatoi</span> found in cat feces recovered by the centrifugal flotation technique, 400× magnification: (<b>A</b>) from cat café no. 03 (<b>B</b>) from cat café no. 06.</p>
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<p>Example of dermatophytes cultured from hair samples of cats residing in cat cafés: (<b>A</b>) The growth of typical white fluffy colony on DTM media that changed to purple color after 14 days incubation. (<b>B</b>) Several spindle-shaped macroconidia of <span class="html-italic">M. canis</span> stained in lactophenol cotton blue from subculture on SDA for 21 days, 400× magnification.</p>
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15 pages, 844 KiB  
Review
Host-Pathogen Interaction and Resistance Mechanisms in Dermatophytes
by Eleonora Dubljanin, Jelena Zunic, Isidora Vujcic, Ivana Colovic Calovski, Sandra Sipetic Grujicic, Stefan Mijatovic and Aleksandar Dzamic
Pathogens 2024, 13(8), 657; https://doi.org/10.3390/pathogens13080657 - 4 Aug 2024
Viewed by 772
Abstract
Dermatophytes are widely distributed in the environment, with an estimated prevalence of 20–25% of the the global population yearly. These fungi are keratinophilic and keratinolytic and cause the infection of keratin-rich structures such as skin, hair, and nails. The pattern of this infectious [...] Read more.
Dermatophytes are widely distributed in the environment, with an estimated prevalence of 20–25% of the the global population yearly. These fungi are keratinophilic and keratinolytic and cause the infection of keratin-rich structures such as skin, hair, and nails. The pattern of this infectious disease covers a wide spectrum from exposed individuals without symptoms to those with acutely inflammatory or non-inflammatory, chronic to invasive, and even life-threatening symptoms. This review summarizes current information on the pathogenicity, virulence factors, and drug resistance mechanisms associated with dermatophytes. A greater number of virulence factors of these fungi are important for the occurrence of infection and the changes that occur, including those regarding adhesins, the sulfite efflux pump, and proteolytic enzymes. Other virulence factors include mechanisms of evading the host defense, while the development of resistance to antifungal drugs is increasing, resulting in treatment failure. The investigation of host-pathogen interactions is essential for developing a more complete understanding of the mechanisms underlying dermatophyte pathogenesis and host response to inform the use of diagnostics methods and antifungal therapeutics to minimize the high fungal burden caused by dermatophytes and to control the spread of resistance. Full article
(This article belongs to the Section Fungal Pathogens)
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<p>Disease pyramid in dermatophytosis.</p>
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<p>Schematic representation of the steps in the degradation of keratinous structures by dermatophytes.</p>
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12 pages, 8155 KiB  
Article
Caffeine Protects Keratinocytes from Trichophyton mentagrophytes Infection and Behaves as an Antidermatophytic Agent
by Diogo M. da Fonseca, Lisa Rodrigues, José Sousa-Baptista, Félix Marcos-Tejedor, Marta Mota, Rodrigo A. Cunha, Chantal Fernandes and Teresa Gonçalves
Int. J. Mol. Sci. 2024, 25(15), 8303; https://doi.org/10.3390/ijms25158303 - 30 Jul 2024
Viewed by 549
Abstract
Caffeine affords several beneficial effects on human health, acting as an antioxidant, anti-inflammatory agent, and analgesic. Caffeine is widely used in cosmetics, but its antimicrobial activity has been scarcely explored, namely against skin infection agents. Dermatophytes are the most common fungal agents of [...] Read more.
Caffeine affords several beneficial effects on human health, acting as an antioxidant, anti-inflammatory agent, and analgesic. Caffeine is widely used in cosmetics, but its antimicrobial activity has been scarcely explored, namely against skin infection agents. Dermatophytes are the most common fungal agents of human infection, mainly of skin infections. This work describes the in vitro effect of caffeine during keratinocyte infection by Trichophyton mentagrophytes, one of the most common dermatophytes. The results show that caffeine was endowed with antidermatophytic activity with a MIC, determined following the EUCAST standards, of 8 mM. Caffeine triggered a modification of the levels of two major components of the fungal cell wall, β-(1,3)-glucan and chitin. Caffeine also disturbed the ultrastructure of the fungal cells, particularly the cell wall surface and mitochondria, and autophagic-like structures were observed. During dermatophyte–human keratinocyte interactions, caffeine prevented the loss of viability of keratinocytes and delayed spore germination. Overall, this indicates that caffeine can act as a therapeutic and prophylactic agent for dermatophytosis. Full article
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<p>Effect of caffeine on β-(1,3)-glucan and chitin levels of <span class="html-italic">T. mentagrophytes</span> cell wall. <span class="html-italic">T. mentagrophytes</span> microconidia were inoculated in YME supplemented with 1, 5, or 10 mM of caffeine prior to the quantification of (<b>A</b>) β-(1,3)-glucan or (<b>B</b>) chitin levels in the fungal cell wall. Under the control condition, the fungus was grown in YME containing no caffeine. Results are presented as mean ± SEM (<span class="html-italic">n</span> = 3); * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 with an unpaired <span class="html-italic">t</span>-test.</p>
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<p>Impact of caffeine on <span class="html-italic">T. mentagrophytes</span> ultrastructure. Fungal microconidia were inoculated in YME in the absence or presence of 10 mM caffeine. Acquired images from (<b>A</b>–<b>D</b>) represent fungi grown in the absence of caffeine, and images from (<b>E</b>–<b>L</b>) represent fungi grown in media supplemented with 10 mM caffeine. “v” identifies vacuoles, “n” nuclei.</p>
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<p>Impact of caffeine on <span class="html-italic">T. mentagrophytes</span> ultrastructure. Fungal microconidia were inoculated in YME in the absence or presence of 10 mM caffeine. Acquired images from (<b>A</b>–<b>D</b>) represent fungi grown in the absence of caffeine, and images from (<b>E</b>–<b>L</b>) represent fungi grown in media supplemented with 10 mM caffeine. “v” identifies vacuoles, “n” nuclei.</p>
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<p>Effect of caffeine on HaCaT cell viability and impact during an in vitro infection with <span class="html-italic">T. mentagrophytes</span> (Tm) microconidia. Keratinocytes were exposed to caffeine (caf.) at different concentrations for 6, 9, and 12 h. Cell viability was quantified using the MTT assay. Under control condition, no caffeine was added to cells. (<b>A</b>) The viability of HaCaT cell was determined in the absence of fungal microconidia and (<b>B</b>) in the presence of fungal <span class="html-italic">T. mentagrophytes</span> spores. Results are presented as mean ± SEM (<span class="html-italic">n</span> = 3); * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001 (vs. control in (<b>A</b>) and vs. Tm in (<b>B</b>)) using two-way ANOVA.</p>
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<p>Effect of caffeine on <span class="html-italic">T. mentagrophytes</span> microconidia germination during the course of an in vitro infection. HaCaT cells were infected with <span class="html-italic">T. mentagrophytes</span> microconidia in the absence (control condition) or in the presence of 10 mM caffeine. After each incubation period, the number of ungerminated microconidia was counted. The number of ungerminated microconidia in control conditions (no caffeine) was normalized to 1. Results are presented relative to control as mean ± SEM (<span class="html-italic">n</span> = 3).</p>
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38 pages, 29521 KiB  
Article
Bioinformatics-Driven mRNA-Based Vaccine Design for Controlling Tinea Cruris Induced by Trichophyton rubrum
by Amir Elalouf, Hanan Maoz and Amit Yaniv Rosenfeld
Pharmaceutics 2024, 16(8), 983; https://doi.org/10.3390/pharmaceutics16080983 - 25 Jul 2024
Viewed by 764
Abstract
Tinea cruris, a dermatophyte fungal infection predominantly caused by Trichophyton rubrum and Epidermophyton floccosum, primarily affects the groin, pubic region, and adjacent thigh. Its recurrence is frequent, attributable to repeated fungal infections in susceptible individuals, especially those with onychomycosis or tinea pedis, [...] Read more.
Tinea cruris, a dermatophyte fungal infection predominantly caused by Trichophyton rubrum and Epidermophyton floccosum, primarily affects the groin, pubic region, and adjacent thigh. Its recurrence is frequent, attributable to repeated fungal infections in susceptible individuals, especially those with onychomycosis or tinea pedis, which act as reservoirs for dermatophytes. Given the persistent nature of tinea cruris, vaccination emerges as a promising strategy for fungal infection management, offering targeted, durable protection against various fungal species. Vaccines stimulate both humoral and cell-mediated immunity and are administered prophylactically to prevent infections while minimizing the risk of antifungal resistance development. Developing fungal vaccines is challenging due to the thick fungal cell wall, similarities between fungal and human cells, antigenic variation, and evolutionary resemblance to animals, complicating non-toxic target identification and T-cell response variability. No prior research has shown an mRNA vaccine for T. rubrum. Hence, this study proposes a novel mRNA-based vaccine for tinea cruris, potentially offering long-term immunity and reducing reliance on antifungal medications. This study explores the complete proteome of T. rubrum, identifying potential protein candidates for vaccine development through reverse vaccinology. Immunogenic epitopes from these candidates were mapped and integrated into multitope vaccines and reverse translated to construct mRNA vaccines. Then, the mRNA was translated and computationally assessed for physicochemical, chemical, and immunological attributes. Notably, 1,3-beta-glucanosyltransferase, CFEM domain-containing protein, cell wall galactomannoprotein, and LysM domain-containing protein emerged as promising vaccine targets. Antigenic, immunogenic, non-toxic, and non-allergenic cytotoxic T lymphocyte, helper T lymphocyte, and B lymphocyte epitopes were selected and linked with appropriate linkers and Toll-like receptor (TLR) agonist adjuvants to formulate vaccine candidates targeting T. rubrum. The protein-based vaccines underwent reverse translation to construct the mRNA vaccines, which, after inoculation, were translated again by host ribosomes to work as potential components for triggering the immune response. After that, molecular docking, normal mode analysis, and molecular dynamic simulation confirmed strong binding affinities and stable complexes between vaccines and TLR receptors. Furthermore, immune simulations of vaccines with and without adjuvant demonstrated activation of immune responses, evidenced by elevated levels of IgG1, IgG2, IgM antibodies, cytokines, and interleukins. There was no significant change in antibody production between vaccines with and without adjuvants, but adjuvants are crucial for activating the innate immune response via TLRs. Although mRNA vaccines hold promise against fungal infections, further research is essential to assess their safety and efficacy. Experimental validation is crucial for evaluating their immunogenicity, effectiveness, and safety. Full article
(This article belongs to the Special Issue Trends in mRNA Vaccine Development and Applications)
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<p>Predicted 3D structures of mRNA-derived vaccine candidates BGTV (<b>a</b>), CDPV (<b>b</b>), GMPV (<b>c</b>), and LDPV (<b>d</b>). Generated using trRosettaRNA.</p>
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<p>Visual representation of secondary structures of BGTV (<b>a</b>), CDPV (<b>b</b>), GMPV (<b>c</b>), and LDPV (<b>d</b>).</p>
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<p>Superimposed 3D models of unrefined (purple) and refined (green) BGTV (<b>a</b>), CDPV (<b>c</b>), GMPV (<b>e</b>), and LDPV (<b>g</b>) with Ramachandran plots of refined 3D constructs of BGTV (<b>b</b>), CDPV (<b>d</b>), GMPV (<b>f</b>), and LDPV (<b>h</b>).</p>
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<p>Docking complexes of vaccine candidates (red) against <span class="html-italic">T. rubrum</span> and TLR2 receptor (blue). (<b>a</b>) BGTV-TLR2; (<b>b</b>) CDPV-TLR2; (<b>c</b>) GMPV-TLR2; (<b>d</b>) LDPV-TLR2.</p>
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<p>Docking complexes of vaccine candidates (red) against <span class="html-italic">T. rubrum</span> and TLR4 receptor (blue). (<b>a</b>) BGTV-TLR4; (<b>b</b>) CDPV-TLR4; (<b>c</b>) GMPV-TLR4; (<b>d</b>) LDPV-TLR4.</p>
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<p>Normal mode analysis (NMA) of vaccine candidates against <span class="html-italic">T. rubrum</span> and TLR2 receptor complexes by iMODs. (<b>a</b>–<b>e</b>) iMODS results of BGTV-TLR2 complex. (<b>a</b>) NMA mobility; (<b>b</b>) main-chain deformability; (<b>c</b>) B-factor values; (<b>d</b>) the eigenvalue; (<b>e</b>) variance. (<b>f</b>–<b>j</b>) iMODS results of CDPV-TLR2 complex. (<b>f</b>) NMA mobility; (<b>g</b>) main-chain deformability; (<b>h</b>) B-factor values; (<b>i</b>) the eigenvalue; (<b>j</b>) variance; (<b>k</b>–<b>o</b>) iMODS results of GMPV-TLR2 complex. (<b>k</b>) NMA mobility; (<b>l</b>) main-chain deformability; (<b>m</b>) B-factor values; (<b>n</b>) the eigenvalue; (<b>o</b>) variance; (<b>p</b>–<b>t</b>) iMODS results of LDPV-TLR2 complex. (<b>p</b>) NMA mobility; (<b>q</b>) main-chain deformability; (<b>r</b>) B-factor values; (<b>s</b>) the eigenvalue; (<b>t</b>) variance.</p>
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<p>Normal mode analysis (NMA) of vaccine candidates against <span class="html-italic">T. rubrum</span> and TLR4 receptor complexes by iMODs. (<b>a</b>–<b>e</b>) iMODS results of BGTV-TLR24 complex. (<b>a</b>) NMA mobility; (<b>b</b>) main-chain deformability; (<b>c</b>) B-factor values; (<b>d</b>) the eigenvalue; (<b>e</b>) variance. (<b>f</b>–<b>j</b>) iMODS results of CDPV-TLR4 complex. (<b>f</b>) NMA mobility; (<b>g</b>) main-chain deformability; (<b>h</b>) B-factor values; (<b>i</b>) the eigenvalue; (<b>j</b>) variance; (<b>k</b>–<b>o</b>) iMODS results of GMPV-TLR4 complex. (<b>k</b>) NMA mobility; (<b>l</b>) main-chain deformability; (<b>m</b>) B-factor values; (<b>n</b>) the eigenvalue; (<b>o</b>) variance; (<b>p</b>–<b>t</b>) iMODS results of LDPV-TLR4 complex. (<b>p</b>) NMA mobility; (<b>q</b>) main-chain deformability; (<b>r</b>) B-factor values; (<b>s</b>) the eigenvalue; (<b>t</b>) variance.</p>
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<p>MD simulation results of dock complexes of potential vaccine candidates (BGTV (black), CDPV (blue), GMPV (yellow), and LDPV (red)) with TLR2 backbone. (<b>a</b>) Trajectory analysis of the RMSD between C-alpha atoms of dock complexes over time, (<b>b</b>) RMSF plot, (<b>c</b>) number of hydrogen bond formations, and (<b>d</b>) radius of gyration (RoG) plot.</p>
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<p>MD simulation results of dock complexes of potential vaccine candidates (BGTV (black), CDPV (blue), GMPV (yellow), and LDPV (red)) with TLR4 backbone. (<b>a</b>) Trajectory analysis of the RMSD between C-alpha atoms of dock complexes over time, (<b>b</b>) RMSF plot, (<b>c</b>) number of hydrogen bond formations, and (<b>d</b>) radius of gyration (RoG) plot.</p>
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<p>A computer-based simulation to model the immune response to the BGTV candidate, administering three doses over 350 days. Key parameters evaluated included antigen and immunoglobulins levels (<b>a</b>), LBLs (<b>b</b>–<b>d</b>), HTLs and CTLs (<b>e</b>–<b>i</b>), natural killer cells (<b>j</b>), dendritic cells (<b>k</b>), macrophages (<b>l</b>), epithelial presenting cell population (<b>m</b>), and cytokine concentrations (<b>n</b>). The Simpson index (D) was utilized to assess the simulation outcomes.</p>
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<p>A computer-based simulation to model the immune response to the CDPV candidate, administering three doses over 350 days. Key parameters evaluated included antigen and immunoglobulins levels (a), LBLs (b–d), HTLs and CTLs (e–i), natural killer cells (j), dendritic cells (k), macrophages (l), epithelial presenting cell population (m), and cytokine concentrations (n). The Simpson index (D) was utilized to assess the simulation outcomes.</p>
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<p>A computer-based simulation to model the immune response to the GMPV candidate, administering three doses over 350 days. Key parameters evaluated included antigen and immunoglobulins levels (a), LBLs (b–d), HTLs and CTLs (e–i), natural killer cells (j), dendritic cells (k), macrophages (l), epithelial presenting cell population (m), and cytokine concentrations (n). The Simpson index (D) was utilized to assess the simulation outcomes.</p>
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<p>A computer-based simulation to model the immune response to the LDPV candidate, administering three doses over 350 days. Key parameters evaluated included antigen and immunoglobulins levels (a), LBLs (b–d), HTLs and CTLs (e–i), natural killer cells (j), dendritic cells (k), macrophages (l), epithelial presenting cell population (m), and cytokine concentrations (n). The Simpson index (D) was utilized to assess the simulation outcomes.</p>
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13 pages, 626 KiB  
Article
New Diagnostic Strategy for Onychomycosis: First-Line Utilization of DermaGenius® PCR and Calcofluor Microscopy Combined with Selective Culturing
by Séverine Evrard, Caroline Minon, Mouhsine Lamtiri Laarif, Benjamin De Backer, Henry Paridaens, Marie-Pierre Hayette, Julie Frère, Jean-Marc Senterre and Jean-Marc Minon
J. Fungi 2024, 10(8), 515; https://doi.org/10.3390/jof10080515 - 24 Jul 2024
Viewed by 589
Abstract
Onychomycosis (OM) is a widespread infection requiring prolonged treatment with potential side effects. Diagnostic certainty is therefore essential before initiating antifungal therapy. Molecular biology has already shown benefits in reducing the time to diagnosis, providing technical ease, and increasing sensitivity for the respective [...] Read more.
Onychomycosis (OM) is a widespread infection requiring prolonged treatment with potential side effects. Diagnostic certainty is therefore essential before initiating antifungal therapy. Molecular biology has already shown benefits in reducing the time to diagnosis, providing technical ease, and increasing sensitivity for the respective species that molecular tests can detect. Nevertheless, causative agents are numerous, and culture remains essential, particularly for detecting non-dermatophytes mold infections. This study compared the performance of three different diagnostic strategies: conventional culture technique, the multiplex DermaGenius® 2.0 PCR (DG), and a mixed PCR/culture algorithm guided by the result of direct examination with calcofluor (DEC). The mixed algorithm (MA) prioritizes DG PCR and DEC as the primary diagnostic tools, supplemented by selective sample inoculation when mycelial elements are visualized in DEC and when DG PCR fails to detect any fungus or identifies a fungus with morphology differing from that observed in DEC (filamentous fungi versus yeasts). With only 13% of samples requiring inoculation, MA emerged as the most effective strategy, demonstrating significantly higher sensitivity (98.18%; p < 0.001) compared to single-method approaches (78.18% for DG PCR alone and 74.55% for culture alone) while maintaining a specificity comparable to DG PCR (100%). This new approach saves time in result delivery, requires fewer human resources, and increases diagnostic accuracy to better meet the needs of clinicians. Full article
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<p>Distribution of fungus type according to the patient’s age.</p>
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13 pages, 3364 KiB  
Article
Synergistic Antimicrobial Activity of Biogenic Silver Nanoparticles and Acanthospermum australe Essential Oil against Skin Infection Pathogens
by Javier Mussin and Gustavo Giusiano
Antibiotics 2024, 13(7), 674; https://doi.org/10.3390/antibiotics13070674 - 20 Jul 2024
Viewed by 713
Abstract
In response to the steady increase in antimicrobial-resistant strains, the World Health Organisation has emphasised the need to investigate new antimicrobial agents and alternative therapies that improve the spectrum of activity and reduce the dose required, thus improving safety. This study focused on [...] Read more.
In response to the steady increase in antimicrobial-resistant strains, the World Health Organisation has emphasised the need to investigate new antimicrobial agents and alternative therapies that improve the spectrum of activity and reduce the dose required, thus improving safety. This study focused on the characterisation of Acanthospermum australe essential oil and green-synthesis silver nanoparticles (AgNP), evaluating their cytotoxicity in human cells, antimicrobial activity and synergistic effect against pathogens causing skin infections. The main components of the essential oil were germacrene A (24.07%), γ-cadinene (21.47%) and trans-caryophyllene (14.97%). Spherical AgNP with a diameter of 15 ± 3 nm were synthesised. The essential oil showed antimicrobial activity against dermatophytes and Malassezia globosa, while AgNP were found to be active against bacteria, yeasts and dermatophytes. Both compounds were found to be primarily non-cytotoxic at the concentrations required to inhibit microbial growth. Furthermore, the combined use of essential oil and AgNP showed a synergistic antimicrobial effect against dermatophytes and M. globosa. In conclusion, the results suggest that the combined use of bioactive compounds from natural sources, such as essential oil and biogenic AgNP, has the potential to improve antimicrobial efficacy against specific skin pathogens, particularly Microsporum canis, Nannizzia gypsea and M. globosa. Full article
(This article belongs to the Special Issue Silver and Gold Compounds as Antibiotics, 2nd Edition)
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<p>(<b>a</b>) UV–visible spectrum, (<b>b</b>) FTIR, (<b>c</b>) TEM image and (<b>d</b>) EDX of the synthesised AgNP.</p>
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<p>Percentage of cell viability after 24 h of incubation as a function of the logarithm of the concentration (µg/mL) of essential oil and AgNP. Values tested: 0.01 = log 1.024; 1.01 = log 10.24; 2.01 = log 102.4; 3.01 = log 1024. The dashed red line indicates 70% viability. Samples with less than 70% viability were considered cytotoxic.</p>
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<p>Heat map constructed from the MIC values obtained for the essential oil and AgNP against the tested microorganisms. EO: essential oil. Drug controls: Itraconazole for dermatophytes and yeasts, penicillin for S. pyogenes and gentamicin for other bacteria.</p>
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26 pages, 5605 KiB  
Article
Ocimum basilicum and Lagenaria siceraria Loaded Lignin Nanoparticles as Versatile Antioxidant, Immune Modulatory, Anti-Efflux, and Antimicrobial Agents for Combating Multidrug-Resistant Bacteria and Fungi
by Lamiaa A. El-Samahy, Yasmine H. Tartor, Adel Abdelkhalek, Ioan Pet, Mirela Ahmadi and Sameh M. El-Nabtity
Antioxidants 2024, 13(7), 865; https://doi.org/10.3390/antiox13070865 - 19 Jul 2024
Viewed by 810
Abstract
Lignin nanoparticles emerged as a promising alternative for drug delivery systems owing to their biodegradability and bioactive properties. This study investigated the antimicrobial activity of the ethanolic extract of Ocimum basilicum-loaded lignin nanoparticles (OB-LNPs) and Lagenaria siceraria seed oil-loaded lignin nanoparticles (LS-LNPs) [...] Read more.
Lignin nanoparticles emerged as a promising alternative for drug delivery systems owing to their biodegradability and bioactive properties. This study investigated the antimicrobial activity of the ethanolic extract of Ocimum basilicum-loaded lignin nanoparticles (OB-LNPs) and Lagenaria siceraria seed oil-loaded lignin nanoparticles (LS-LNPs) to find a solution for antimicrobial resistance. OB-LNPs and LS-LNPs were tested for their antimicrobial potential against Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Staphylococcus aureus, Salmonella enterica, Trichophyton mentagrophytes, Trichophyton rubrum, and Microsporum canis. OB-LNPs and LS-LNPs were further tested for their anti-efflux activity against ciprofloxacin-resistant Salmonella enterica strains and for treating Salmonella infection in a rat model. We also investigated the antifungal efficacy of OB-LNPs and LS-LNPs for treating T. rubrum infection in a guinea pig model. Both OB-LNPs and LS-LNPs showed strong antimicrobial potential against S. Typhimurium and T. rubrum infections. LS-LNPs showed antibacterial activity against Salmonella enterica species with a MIC range of 0.5–4 µg/mL and antifungal activity against T. rubrum with a MIC range of 0.125–1 µg/mL. OB-LNPs showed antibacterial activity against Salmonella enterica species with a MIC range of 0.5–2 µg/mL and antifungal activity against T. rubrum with a MIC range of 0.25–2 µg/mL. OB-LNPs and LS-LNPs downregulated the expression of ramA and acrB efflux pump genes (fold change values ranged from 0.2989 to 0.5434; 0.4601 to 0.4730 for ramA and 0.3842–0.6199; 0.5035–0.8351 for acrB). Oral administration of OB-LNPs and LS-LNPs in combination with ciprofloxacin had a significant effect on all blood parameters, as well as on liver and kidney function parameters. Oxidative stress mediators, total antioxidant capacity, and malondialdehyde were abolished by oral administration of OB-LNPs and LS-LNPs (0.5 mL/rat once daily for 5 days). Interferon-γ and tumor necrosis factor-α were also reduced in comparison with the positive control group and the ciprofloxacin-treated group. Histopathological examination of the liver and intestine of OB-LNPs and LS-LNPs-treated rats revealed an elevation in Salmonella clearance. Treatment of T. rubrum-infected guinea pigs with OB-LNPs and LS-LNPs topically in combination with itraconazole resulted in a reduction in lesion scores, microscopy, and culture results. In conclusion, OB-LNPs and LS-LNPs possess immunomodulatory and antioxidant potential and can be used as naturally derived nanoparticles for drug delivery and treatment of Salmonellosis and dermatophytosis infections. Full article
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<p>Schematic illustration of the green synthesis of LS-LNPs and OB-LNPs.</p>
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<p>TEM micrographs and selected area electron diffraction (SAED) patterns of LS-LNPs (<b>A</b>) and OB-LNPs (<b>B</b>) showing nano sphere shape with concentric rings of brilliant spots.</p>
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<p>UV-visible spectra (<b>A</b>), FT-IR spectra (<b>B</b>), zeta potential (<b>C</b>), and XRD patterns (<b>D</b>) of LS-LNPs and OB-LNPs.</p>
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<p>Expression of <span class="html-italic">ram</span>A and <span class="html-italic">acr</span>B efflux genes in <span class="html-italic">Salmonella enterica</span> strains treated with OB-LNPs and LS-LNPs. The transcription of the <span class="html-italic">ram</span>A and <span class="html-italic">acr</span>B genes significantly reduced in response to treatment with OB-LNPs and LS-LNPs, minimizing the OB-LNPs. ** indicates a highly significant difference at <span class="html-italic">p</span> value &lt; 0.001.</p>
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<p>Total bacterial counts (Log<sub>10</sub> CFU) in liver (<b>A</b>), spleen (<b>B</b>), and intestine (<b>C</b>) samples from rats in different groups treated with CIP (45 mg/kg bw), Cip+OB-LNPs (45 mg/kg bw + 2.5 mL/kg bw), Cip+LS-LNPs (45 mg/kg bw + 2.5 mL/kg bw), OB-LNPs (2.5 mL/kg bw), and LS-LNPs (2.5 mL/kg bw), daily for 5 days post <span class="html-italic">S</span>. Typhimurium challenge.</p>
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<p>Photomicrograph from the ileum and jejunum of rats using H&amp;E staining at 7th day post infection with <span class="html-italic">S</span>. Typhimurium. (<b>A</b>) noninfected and nontreated rats showing normal mucosal villi comprising absorptive columnar epithelium with variable number of goblet cells (black arrow), normal Peyer’s patch (yellow arrow), normal submucosa (light blue arrow), and muscular coat (green arrow). (<b>B</b>) infected, non-treated rats showing villous necrosis (black arrows), mucosal leucocytic infiltration (green arrows), and moderate lymphoid hyperplasia of Peyer’s patch (yellow arrows). (<b>C</b>) rats were infected and treated with 45 mg/kg CIP (ciprofloxacin) orally once daily for 5 days. The ileum revealed normal submucosal glands (black arrow) and mildly reactive Peyer’s patch (yellow arrow). The jejunum revealed normal jejunal mucosal villi comprising absorptive columnar epithelium with a variable number of goblet cells (black arrow), and few lymphocytes are seen in the submucosa (light blue arrow). (<b>D</b>) rats were infected and received CIP+OB-LNPs (45 mg/kg bw + 2.5 mL/kg bw). The jejunum and ileum show normal mucosal villi comprising absorptive columnar epithelium with a variable number of goblet cells, occasional lymphocytic infiltration (black arrow), and normal Peyer’s patch (yellow arrow), beside normal submucosa (light blue arrows) and muscular coat (green arrow). (<b>E</b>) rats were infected and treated with CIP+LS-LNPs (45 mg/kg bw + 2.5 mL/kg bw) showing normal mucosal villi comprising absorptive columnar epithelium with variable number of goblet cells (black arrow) and normal Peyer’s patch (yellow arrow), beside normal submucosa (light blue arrows) and muscular coat (green arrow). Ileal villi of some sections appear moderately stunted (white stars). (<b>F</b>) rats were infected and treated with OB-LNPs (2.5 mL/kg bw), showing normal mucosal villi comprising absorptive columnar epithelium with a variable number of goblet cells and occasional epithelial stratification (black arrow), moderately reactive Peyer’s patch (yellow arrow), normal submucosa (light blue arrows), and muscular coat (green arrow). (<b>G</b>) rats were infected and treated with LS-LNPs (2.5 mL/kg bw) daily for 5 days, showing normal mucosal villi with variable number of goblet cells (black arrow), normal Peyer’s patch (yellow arrow), normal submucosa (light blue arrows), and muscular coat (green arrow).</p>
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<p>Photomicrographs of liver sections of rats using H&amp;E staining at 7th day post infection with <span class="html-italic">S</span>. Typhimurium. (<b>A</b>) noninfected and nontreated rats showing a normal central vein and portal area comprising vascular and biliary structures (red arrow at 120 µm; orange and blue arrows at 60 µm) and, hepatic sinusoids (black star), and hepatocytes (red star). (<b>B</b>) infected non treated rats showing focal hepato-cellular degeneration (green arrowhead), early necrotic changes (yellow arrowhead), and scattered apoptosis (yellow star) beside focal round cells aggregation (dark green arrow). (<b>C</b>) rats were infected and treated with 45 mg/kg CIP (ciprofloxacin) orally once daily for 5 days, showing mildly congested portal blood vessels and biliary proliferation (yellow star and red arrow). Normal hepatic sinusoids (black star) and hepatocytes (red star) are also seen. (<b>D</b>) rats were infected and received CIP+OB-LNPs (45 mg/kg bw + 2.5 mL/kg bw) showing apparently normal hepatic parenchyma with occasional portal fibroplasia (brown star), biliary proliferation (orange star), reactive Von-Kupffer cells (black star), and focal interstitial round cells aggregation (dark green arrow). (<b>E</b>) rats were infected and treated with CIP+ LS-LNPs (45 mg/kg bw + 2.5 mL/kg bw), showing normal central vein (red arrow), portal area comprising vascular and biliary structures (orange and blue arrows), hepatic sinusoids (black star), and hepatocytes (red star). (<b>F</b>) rats were infected and treated with OB-LNPs (2.5 mL/kg bw), showing apparently normal hepatic parenchyma (red arrow, red star, and dark blue arrows) with occasional reactive Von-Kupffer cells (black star) and focal interstitial round cells aggregation (dark green arrow). (<b>G</b>) rats were infected and treated with LS-LNPs (2.5 mL/kg bw) daily for 5 days showing apparently normal hepatic parenchyma (red arrow, red star, and dark blue arrows) with occasional reactive Von-Kupffer cells (black star).</p>
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<p>Means of erythema (<b>A</b>), scales (<b>B</b>), and alopecia (<b>C</b>) scores in <span class="html-italic">Trichophyton rubrum</span> infected guinea pigs in different groups. Symbols on the graphs indicate significant differences among the treated groups (itraconazole (ITRA), ITRA+LC-LNPs, ITRA+OB-LNPs, OB-LNPs, and LS-LNPs) and the untreated positive control group (<span class="html-italic">p</span> &lt; 0.05). <span class="html-italic">p</span>-value was estimated using Exact Wilcoxon test.</p>
Full article ">Figure 8 Cont.
<p>Means of erythema (<b>A</b>), scales (<b>B</b>), and alopecia (<b>C</b>) scores in <span class="html-italic">Trichophyton rubrum</span> infected guinea pigs in different groups. Symbols on the graphs indicate significant differences among the treated groups (itraconazole (ITRA), ITRA+LC-LNPs, ITRA+OB-LNPs, OB-LNPs, and LS-LNPs) and the untreated positive control group (<span class="html-italic">p</span> &lt; 0.05). <span class="html-italic">p</span>-value was estimated using Exact Wilcoxon test.</p>
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