Combined mutations of the penA with ftsX genes contribute to ceftriaxone resistance in Neisseria gonorrhoeae and peptide nucleic acids targeting these genes reverse ceftriaxone resistance.

OBJECTIVES: To investigate the gene mutations associated with ceftriaxone (CRO) resistance among gonococcal isolates, and to determine the effects of the mutated genes on CRO minimum inhibitory concentrations (MICs) with transformation assays and antisense peptide nucleic acids (asPNAs). METHODS: Ceftriaxone-resistant (CROR) and ceftriaxone-susceptible (CROS) isolates were identified using EUCAST and paired according to similarity in their MICs to other antimicrobials. The two groups of gonococci were sequenced and analysed. Mutated genes that showed a statistical difference between the two groups were transformed into gonococcal reference strains to determine their functions. AsPNAs were designed and transformed into the former transformant to further confirm the effects of the mutated genes. RESULTS: Twenty-two paired CROR and CROS isolates were obtained. The incidence of the penA-A501T and penA-G542S mutations individually, as well as combined mutations (penA-A501T and ftsX-R251H, penA-G542S and ftsX R251H), was statistically different between the two groups. The MIC of ATCC43069 (A43) increased 2 times following transformation with penA-A501T, and the MICs of A43 and ATCC49226 (A49) increased 32 times and 2 times following transformation with penA-A501T and ftsX-R251H, respectively. Antisense PNA-P3 reduced the MIC of the A43 transformant most significantly when transformed individually. PNA-P3 and PNA-F1 (asPNAs of the penA and ftsX) restored CRO susceptibility. CONCLUSIONS: PenA-A501T and penA-G542S mutations are important in CRO resistance among gonococci isolates. The ftsX-R251H mutation is also related to CRO resistance, and combined mutations of ftsX-R251H and penA-A501T comediate a significant reduction in CRO susceptibility. The combined application of PNA-P3 and PNA-F1 could effectively reverse the resistance to CRO in N. gonorrhoeae.

Exploring the clinical features and risk factors for children tinea capitis complicated with allergic diseases.

BACKGROUND: Tinea capitis, atopic dermatitis and allergic rhinitis are the most common disorders endured by prepubescent children. Dermatophyte infections have been linked to allergic disorders, such as increased sensitivity to dermatophytes in patients with atopic dermatitis. OBJECTIVES: To explore the correlation between tinea capitis and allergic diseases in children and to analyse their risk factors. METHODS: This study monitored epidemiological changes in childhood tinea capitis and risk factors for whom with allergic disease in a single centre in three consecutive five-year intervals by reviewing clinical data and multivariate logistic data analysis. RESULTS: Between 2007 and 2022, there were 127 children patients with tinea capitis, the mean age was 4.83 years, and the male-to-female ratio was 1.76:1. Zoophilic Microsporum canis and Trichophyton mentagrophytes were the most prevalent pathogens, and the proportions remained relatively constant every 5 years. There were 34 (26.8%) children with tinea capitis complicated with allergic disease, among them 14 children with atopic dermatitis/eczema, 13 with allergic rhinitis, 8 urticaria, 6 food allergies and 1 allergic asthma. Male, kerion, zoophilic species infections and animal contact history were prevalent features in allergic disease combined with tinea capitis. Patients with tinea capitis plus allergic disease mostly had a family history with similar complications. CONCLUSION: M. canis and T. mentagrophytes were the most prevalent pathogens of tinea capitis in the last 15 years; atopic dermatitis/eczema and allergic rhinitis were the most frequently associated allergic diseases. Male, kerion, zoophilic pathogen and animal contact history are risk factors.

Human and mouse TLR2 results in different activation of p38 and JNK signal pathway in HaCaT infected by Trichophyton rubrum and Microsporum canis.

Introduction: It has long been recognized that inflammation to dermatophyte infection is different among various hosts, but the mechanism underlying is still not well understood. Toll-like receptor (TLR2), mediates the innate immune response against dermatophyte infection and is very important to trigger the inflammatory response to dermatophytes. Considering the different amino acid sequences and structures of TLR2, we speculated that TLR2 from different hosts will activate the downstream signal pathways to varying degrees, resulting in different inflammatory responses to dermatophytes. Methods: In this study, we constructed the mice-human fusion TLR2 expressed HaCaT (mhTLR2-HaCaT) by replacing the extracellular ligand recognition region of human TLR2 with that of the mouse. Then hTLR2-HaCaT cells and mhTLR2-HaCaT cells were infected with T. rubrum and M. canis for 24 h followed by immunoblotting to asses associated proteins of p38 and JNK signal pathway. Results: Compared with that of human TLR2 expressed HaCaT (hTLR2-HaCaT), levels of phosphorylated p38 protein were increased in mhTLR2-HaCaT cells stimulated by T. rubrum for 24 h, and levels of phosphorylatedJNK and c-Jun protein were increased in mhTLR2-HaCaT cells whenstimulated with M. canis for 24 h. Discussion: Compared with hTLR2-HaCaT cells, p38 and JNK signal pathwayswere activated in mhTLR2-HaCaT after being infected by Trichophyton rubrumand Microsporum canis, respectively. Since p38 and JNK are the mainpathways that transduce the signal for host recognition of dermatophytes andmediate the downstream inflammatory response, it suggested that theinterspecific difference of TLR2 ectodomain may be one of the reasons for thedifferent inflammatory manifestations between humans and mice infected bythese two dermatophytes. Quite especially, the mouse-derived TLR2extracellular recognition region is more effective in recognizing T. rubrum andM. canis to activate the downstream signal pathways, resulting in a tenserinflammatory response against these two dermatophytes.

The design and application of an automated microscope developed based on deep learning for fungal detection in dermatology.

BACKGROUND: Light microscopy to study the infection of fungi in skin specimens is time-consuming and requires automation. OBJECTIVE: We aimed to design and explore the application of an automated microscope for fungal detection in skin specimens. METHODS: An automated microscope was designed, and a deep learning model was selected. Skin, nail and hair samples were collected. The sensitivity and the specificity of the automated microscope for fungal detection were calculated by taking the results of human inspectors as the gold standard. RESULTS: An automated microscope was built, and an image processing model based on the ResNet-50 was trained. A total of 292 samples were collected including 236 skin samples, 50 nail samples and six hair samples. The sensitivities of the automated microscope for fungal detection in skin, nails and hair were 99.5%, 95.2% and 60%, respectively, and the specificities were 91.4%, 100% and 100%, respectively. CONCLUSION: The automated microscope we developed is as skilful as human inspectors for fungal detection in skin and nail samples; however, its performance in hair samples needs to be improved.

In-vivo full-field measurement of microcirculatory blood flow velocity based on intelligent object identification.

Microcirculation plays a crucial role in delivering oxygen and nutrients to living tissues and in removing metabolic wastes from the human body. Monitoring the velocity of blood flow in microcirculation is essential for assessing various diseases, such as diabetes, cancer, and critical illnesses. Because of the complex morphological pattern of the capillaries, both In-vivo capillary identification and blood flow velocity measurement by conventional optical capillaroscopy are challenging. Thus, we focused on developing an In-vivo optical microscope for capillary imaging, and we propose an In-vivo full-field flow velocity measurement method based on intelligent object identification. The proposed method realizes full-field blood flow velocity measurements in microcirculation by employing a deep neural network to automatically identify and distinguish capillaries from images. In addition, a spatiotemporal diagram analysis is used for flow velocity calculation. In-vivo experiments were conducted, and the images and videos of capillaries were collected for analysis. We demonstrated that the proposed method is highly accurate in performing full-field blood flow velocity measurements in microcirculation. Further, because this method is simple and inexpensive, it can be effectively employed in clinics.

Skin sensitivity evaluation: What could impact the assessment results?

BACKGROUND: For the evaluation results of skin sensitivity, such as clinical parameters, stinging test records and biophysical assessments dates might be impacted by many factors, the influence factors need to be further explored, and the skin sensitivity evaluation process and methodology needed distinction and normalization. In this study, we investigated the changes of sensitive skin indexes and lactic acid stinging test results in different seasons, facial regions, skin photo-type, and living habits. METHODS: Twenty-four healthy subjects had completed this study. Lactic acid stinging test was performed in different seasons. Transepidermal water loss (TEWL), skin hydration, sebum secretion, and pH were measured in an environment-controlled room. Correlations between stinging responses, skin biophysical parameters, and sensitive skin inducements in different seasons were statistically analyzed. RESULTS: Skin TEWL, hydration, sebum secretion, and pH values on different facial parts were various. Two-way correlation analysis between the results of lactic acid stinging test in different seasons and the sensitivity factors showed differences between summer, autumn, and winter. The mean scores of lactic acid stinging test increased in autumn. Linear regression analysis of skin sensitivity factors in type III and type IV photobiology skin found that the frequency of sleeping time and eating spicy food in the past of week could infect the sensitive skin evaluation dates statistically (P < .05). DISCUSSION/CONCLUSIONS: Skin sensitivity assessment results were impacted by seasonal transformation, living habits and customs, and facial regions. These indicted that we should consider above interfering factors when evaluated the skin sensitivity for getting more precise dates.

Synergistic effect of terbinafine and amphotericin B in killing Fonsecaea nubica in vitro and in vivo.

Chromoblastomycosis is a chronic fungal infection. Itraconazole and terbinafine are the most recommended antifungal drugs for chromoblastomycosis, while amphotericin B is not usually recommended. A patient with chromoblastomycosis in our hospital showed poor clinical responses to itraconazole and terbinafine. The fungus isolated from the lesions of this patient was identified as Fonsecaea nubica and numbered zssy0803. In vitro antifungal susceptibilities of F. nubica zssy0803 to terbinafine, amphotericin B, itraconazole, voriconazole and caspofungin were evaluated, as well as the combinations of terbinafine with the other four antifungals. The combined effect of terbinafine and amphotericin B on other 20 clinical F. nubica strains was also evaluated. The minimal inhibitory concentrations of terbinafine, amphotericin B, itraconazole, voriconazole and caspofungin on F. nubica zssy0803 were 0.25 µg/mL, 2 µg/mL, 1 µg/mL, 4 µg/mL and 8 µg/mL, respectively. The combination of terbinafine and amphotericin B showed the lowest fractional inhibitory concentration index of 0.28 to F. nubica zssy0803 in comparison with combinations of terbinafine and the other four antifungal drugs. The combination of terbinafine and amphotericin B was also synergistic for all the other 20 F. nubica strains. Then, the combination of oral terbinafine (500 mg/day) and intralesional injections of amphotericin B (1 mg/mL) was used to treat this patient. After this combined therapy for 25 weeks and terbinafine monotherapy for additional 12 weeks, the patient was cured. These findings indicate for the first time that terbinafine and amphotericin B are synergistic in killing F. nubica both in vitro and in vivo.

Efficacy and safety of 2% supramolecular salicylic acid compared with 5% benzoyl peroxide/0.1% adapalene in the acne treatment: a randomized, split-face, open-label, single-center study.

BACKGROUND: Topical drugs for mild to moderate acne include adapalene (ADA) and benzoyl peroxide(BPO). Supramolecular salicylic acid (SSA), a modified SA preparation, is considered as a new effective therapeutic scheme. OBJECTIVES: To compare the safety and efficacy of 2% supramolecular SA (2% SSA) with 0.01% adapalene plus 5% benzoyl peroxide (5%BPO +0.1%ADA) for treatment of facial acne. MATERIALS AND METHODS: This was an open-label, split face, randomized and single-centre clinical trial. Subjects with mild to moderate acne were enrolled. Two percent SSA cream were randomly applied on one side of the face while 5%BPO +0.1%ADA gel was applied on the opposite side for 28 days. The numbers of acne lesions, along with side effects of the targeted area were evaluated by the investigators at day 0, day 14, and day 28. Skin water content, TEWL and skin lightening indexes were measured at the same time. RESULTS: A total of 31 of acne patients completed the trial. Dates showed that 2% SSA had similar effects to 5%BPO +0.1%ADA in reducing papules/pustules (47.9% vs. 49.8%), non-inflammatory lesions (43.1% vs. 42.7%) and total lesions (44.1% vs. 45.6%; all p > 0.05) at day 28. The skin barrier (skin hydration value and TEWL value), skin brightness (L* value) and erythema (a* values) indicators showed no statistical differences in the left and right sides of the face (p > 0.05). CONCLUSION: This study demonstrated that 2% SSA has a similar efficacy with 5%BPO +0.1%ADA in mild to moderate acne treatment. This might be a useful pilot study that could be used to support further larger clinical trials.

Minimal erythema dose, minimal persistent pigment dose which model for whitening products evaluation is better?

BACKGROUND/AIMS: How to select a suitable method in whitening products evaluation is still under discussion. Here, we compared two different artificial pigmentation models and explored an ideal UV dosage for skin whitening products evaluation model establishment. METHODS: Thirty five healthy volunteers with type IV human skin were recruited and the skin minimal erythema dose (MEDs) and minimal persistent pigment dose (MPPDs) were measured. All volunteers were simultaneously exposed to six increasing doses of radiations from different ultraviolet sources on lower back bilateral flattening area: 95% UVA/5% UVB with the radiating doses of 0.75, 0.94, 1.17, 1.46, 1.83, 2.29 MEDs was used on the left side; meanwhile 99% UVA/1% UVB with radiating doses of 6.0, 7.5, 9.4, 11.7, 14.6, 18.3 MPPDs were used on the right side. Observations and pigmentation measurements were carried out before and after UV radiation for 24 weeks. RESULT: 1.83 MED and 2.29 MED induced medium depth pigmentation by 95% UVA/5% UVB irradiation. 1.83 MED dose causing minimal photo-damage on skin was selected as the most suitable dose. With 99% UVA/1% UVB irradiation, 9.4 MPPD and 11.7 MPPD induced medium depth pigmentation. 9.4 MPPD dose causing minimal photo-damage on skin was selected. CONCLUSION: These findings potentiate advanced understanding of UV model establishment and selection for skin whitening products evaluation as related to dermatopharmacology and dermatotoxicology.

Prevalence of low inflammatory tinea genitalis in southern China.

BACKGROUND: Emergence of highly inflammatory genital dermatophyte infections has been reported from Southeast Asia. In view of this, knowledge of the non-outbreak fungal flora of the genitals is required as a baseline study. OBJECTIVES: We present our 12-year experience in a tertiary clinic with the diagnosis of scrotal fungal infections. METHODS: A retrospective review was performed of patients with a diagnosis of scrotal fungal infections proven by direct microscopy and culture. Clinical, mycological and treatment data were collected. RESULTS: In total, 35 male patients were identified, of which 27 concerned dermatophyte infections and eight were yeasts. Nannizzia gypsea was the most common agent (48.6%), presenting as thick pseudomembraneous lesions limited to the scrotum. Trichophyton rubrum (22.9%) and Epidermophyton floccosum (5.7%) mainly presented erythematous, dry and scaly lesions and involving more sites besides the scrotum. Candida albicans (n = 3), C. glabrata (n = 2), C. guilliermondii (n = 1) and Trichosporon asteroides (n = 1), presented various lesions. Sports, sweating and concurrent tineas are hypothesised as predisposing factors. CONCLUSIONS: The prevalent causative agent of scrotum infections is N. gypsea, but wide species diversity is observed. All infections show mild skin inflammation. It is suggested that this genital fungal flora represents the current situation prior to clonal dermatophyte outbreaks.

A novel RFLP-ARMS TaqMan PCR-based method for detecting the BRAF V600E mutation in melanoma.

To enable the rapid and sensitive screening of the BRAF V600E mutation in clinical samples, a novel method combining restriction fragment length polymorphism (RFLP) analysis with the popular amplification refractory mutation system (ARMS) TaqMan quantitative (qPCR) genotyping method in a single reaction tube was developed. A total of 2 primer pairs were designed to enrich for and genotype the BRAF mutational hotspot (RFLP primers and ARMS primers) and a restriction enzyme was used to remove the wild-type alleles. The analysis revealed that this method detected mutant alleles in mixed samples containing >0.1% mutant sequences. In a survey of 53 melanoma samples, this method detected 21 mutation-positive samples. This novel RFLP-ARMS TaqMan qPCR protocol may prove useful for detecting mutations in clinical samples containing only a small proportion of mutant alleles.

Onychomycosis secondary to onychomadesis: an underdiagnosed manifestation.

Onychomycosis is a rare nail disorder in early childhood, while onychomadesis is a periodic idiopathic, non-inflammatory disease that affects the nail matrix and is common in children especially in those who suffer from viral infections. In this study, we investigated recent cases of onychomycosis subsequent to periods of onychomadesis in children. Sixteen young children (six males, 10 females) with a mean age of 36.5 months were diagnosed with onychomadesis, and 13 of the patients had a history of viral infection prior to nail changes. Direct microscopy of nail scaling was positive in 11 cases (68.8%), and culture was positive in the same number of cases. Four Candida species were isolated: Candida glabrata was the most frequent, found in eight cases (72.7%), while C. albicans, C. parapsilosis and C. tropicalis, each were encountered in a single case. All children were treated successfully with or without topical bifonazole therapy.

Primary localized cutaneous amyloidosis with lichen and poikiloderma-like lesions and an excellent response to systemic acitretin.

Primary localized cutaneous amyloidosis is a skin-limited amyloidosis that does not involve internal organs. It is clinically subclassified into 3 general categories and some rare variants. However, there is considerable overlap within the classification. Though there are a variety of therapeutic measures, the treatment is often unsatisfactory, particularly when the disease is severe and extensive. We describe a rare case of primary localized cutaneous amyloidosis with lichen and poikiloderma-like lesions that showed an excellent response to systemic acitretin.

Primary localized cutaneous amyloidosis with lichen and poikiloderma-like lesions and an excellent response to systemic acitretin

Abstract: Primary localized cutaneous amyloidosis is a skin-limited amyloidosis that does not involve internal organs. It is clinically subclassified into 3 general categories and some rare variants. However, there is considerable overlap within the classification. Though there are a variety of therapeutic measures, the treatment is often unsatisfactory, particularly when the disease is severe and extensive. We describe a rare case of primary localized cutaneous amyloidosis with lichen and poikiloderma-like lesions that showed an excellent response to systemic acitretin.

A case of chromoblastomycosis by Fonsecaea nubica indicating a possible insect route of transmission.

A first auricular case of chromoblastomycosis due to Fonsecaea nubica is reported in a 42-year-old Chinese male. He presented a slightly verrucous, erythematous plaque on his right auricle which had gradually extended over a 10-year period, and the patient reported a history of dog flea sting before onset of the lesions. Diagnosis was based on histopathological and mycological examination of clinical samples, which revealed muriform cells. Identification of the aetiological agent was assessed by morphological characteristics and confirmed at species level by sequencing of the rDNA internal transcribed spacer (ITS). The patient showed marked clinical improvement after 3 months combination therapy with itraconazole and terbinafine. The possible mode of transmission of auricular chromoblastomycosis is discussed.

Ultrafast imaging with anti-aliasing based on optical time-division multiplexing.

Serial time-encoded amplified microscopy (STEAM) is a novel ultrafast imaging technique that is based on space-to-time-to-wavelength mapping. Nevertheless, the technique requires a high-cost electronic digitizer of several tens of gigahertz sampling rate to read out sufficient image information. To acquire a large amount of image information by using a relatively low-sampling-rate electronic digitizer, an anti-aliasing technique based on optical time-division multiplexing is proposed. A 38.88 MHz line-scan imaging system is demonstrated experimentally. By using the proposed anti-aliasing technique, a 20 GS/s sampling rate is achieved by employing a 10 GS/s electronic digitizer. Defects and scratches on the target that were not identifiable originally can be clearly distinguished after using the proposed technique. Numerical analysis shows that the image quality can be improved by 4.16 dB, compared to that not using the anti-aliasing technique and at least 2.3 dB comparing to those obtained by bilinear, bicubic, and nearest-neighbor interpolation and Lanczos resampling techniques.

Effect of Culture Supernatant Derived from Trichophyton Rubrum Grown in the Nail Medium on the Innate Immunity-related Molecules of HaCaT.

BACKGROUND: Trichophyton rubrum is superficial fungi characteristically confined to dead keratinized tissues. These observations suggest that the soluble components released by the fungus could influence the host immune response in a cell in contact-free manner. Therefore, this research aimed to analyze whether the culture supernatant derived from T. rubrum grown in the nail medium could elicit the immune response of keratinocyte effectively. METHODS: The culture supernatants of two strains (T1a, T XHB ) were compared for the ß-glucan concentrations and their capacity to impact the innate immunity of keratinocytes. The ß-glucan concentrations in the supernatants were determined with the fungal G-test kit and protein concentrations with bicinchoninic acid protein quantitative method, then HaCaT was stimulated with different concentrations of culture supernatants by adopting morphological method to select a suitable dosage. Expressions of host defense genes were assessed by quantitative polymerase chain reaction after the HaCaT was stimulated with the culture supernatants. Data were analyzed with one-way analysis of variance, followed by the least significant difference test. RESULTS: The T. rubrum strains (T1a and T XHB ) released ß-glucan of 87.530 ± 37.581 pg/ml and 15.747 ± 6.453 pg/ml, respectively into the media. The messenger RNA (mRNA) expressions of toll-like receptor-2 (TLR2), TLR4, and CARD9 were moderately up-regulated in HaCaT within 6-h applications of both supernatants. HaCaT cells were more responsive to T1a than T XHB . The slight increase of dendritic cells-specific intercellular adhesion molecule 3-grabbing nonintegrin expression was faster and stronger, induced by T1a supernatant than T XHB . The moderate decreases of RNase 7, the slight up-regulations of Dectin-1 and interleukin-8 at the mRNA level were detected only in response to T1a rather than T XHB . After a long-time contact, all the elevated defense genes decreased after 24 h. CONCLUSION: The culture supernatant of T. rubrum could directly and transiently activate the innate immune response of keratinocytes.

Disseminated infection caused by Emmonsia pasteuriana in a renal transplant recipient.

Emmonsia pasteuriana is a thermally dimorphic fungus identified in very few human cases. Here, we report a case of a 43-year-old male renal transplant patient from China presenting with multiple painful skin eruptions on his head, nose and left thigh, later accompanied by respiratory failure. Histopathology of the biopsy collected from the left thigh upper ulcer and occipital nodule both demonstrated chronic inflammation with granuloma formation and yeast-like elements. Emmonsia pasteuriana was cultured from two biopsy specimens and their identity was confirmed by sequencing of the rDNA internal transcribed spacer. The patient in intensive care showed marked clinical improvement with antifungal treatment.

Trichophyton rubrum conidia modulate the expression and transport of Toll-like receptor 2 in HaCaT cell.

Trichophyton rubrum (T. rubrum) represents the most important agent of dermatophytosis in humans. T. rubrum infection causes slight inflammation, and tends to be chronic and recurrent. It is suggested that T. rubrum can modulate the innate immune responses of host cells, which result in the failure of host cells to recognize T. rubrum and initiate effective immune responses. In this study we show how T. rubrum conidia modulate the expression and transport of Toll-like receptor 2 in HaCaT cell. Flow cytometric analysis showed that the surface and total expression of Toll-like receptor 2 were upregulated at the very early stage when keratinocytes were exposed to T. rubrum conidia regardless of the dose, and the upregulation of surface TLR2 was much more significant than that of total TLR2. Moreover, TLR2 expression was suppressed after upregulation in the initial stage of T. rubrum exposure, and the decrease of total TLR2 was earlier than that of surface TLR2. Our results suggest that in the early stage, TLR2 of keratinocytes were upregulated and transported to the cell surface. After then, the expression of TLR2 was suppressed by T. rubrum conidia.