Alginate-Based 3D A549 Cell Culture Model to Study Paracoccidioides Infection.

A three-dimensional (3D) lung aggregate model based on sodium alginate scaffolds was developed to study the interactions between Paracoccidioides brasiliensis (Pb) and lung epithelial cells. The suitability of the 3D aggregate as an infection model was examined using cell viability (cytotoxicity), metabolic activity, and proliferation assays. Several studies exemplify the similarity between 3D cell cultures and living organisms, which can generate complementary data due to the greater complexity observed in these designed models, compared to 2D cell cultures. A 3D cell culture system of human A549 lung cell line plus sodium alginate was used to create the scaffolds that were infected with Pb18. Our results showed low cytotoxicity, evidence of increased cell density (indicative of cell proliferation), and the maintenance of cell viability for seven days. The confocal analysis revealed viable yeast within the 3D scaffold, as demonstrated in the solid BHI Agar medium cultivation. Moreover, when ECM proteins were added to the alginate scaffolds, the number of retrieved fungi was significantly higher. Our results highlight that this 3D model may be promising for in vitro studies of host-pathogen interactions.

Spectral pattern study of citrus black rot caused by Alternaria alternata and selecting optimal wavelengths for decay detection.

Fungal decay is one of the most common diseases that affect postharvest operations and sales of citrus. Sometimes, fungal disease develops and spreads inside the fruit and in the advanced stages of the disease, it appears apparent, so the use of efficient and reliable methods for early detection of the disease is very important. In this study, early detection of citrus black rot disease caused by Alternaria genus fungus was examined using spectroscopy. Jaffa oranges were inoculated with Alternaria alternata. The samples were inspected by spectroscopy (200-1100 nm) in the 1st, 2nd, and 3rd weeks after inoculation. The classification of healthy and infected samples and selection of most important wavelengths were conducted by soft independent modeling of class analogy (SIMCA). The most important wavelengths in the detection of healthy and infected samples of the 1st week were 507, 933, 937, and 950 nm with a classification accuracy of 60%. The most important wavelengths of the 2nd week were 522 and 787 nm with a classification accuracy of 60%. Also, wavelengths of 546, 660, 691, and 839 were found to be effective in the 3rd week with a classification accuracy of 100%.

Rapid diagnosis of Talaromyces marneffei infection assisted by metagenomic next-generation sequencing in a HIV-negative patient.

Talaromyces marneffei (T. marneffei), is an opportunistic pathogenic fungus commonly reported in southeast Asia. T. marneffei infection predominantly occurs in patients with immunodeficiency and can be fatal if diagnosis and treatment were delayed. Conventional diagnosis of T. marneffei infection relies heavily on tissue culture and histologic analysis, which is time consuming and has limited positive rate. Rapid and accurate diagnosis of T. marneffei remains urgent for effective therapy and prevention. This case is the first reported T. marneffei infection in non-HIV patients in north China diagnosed by mNGS. The successful diagnosis of T. marneffei infection assistant by mNGS underlies the potential of this technique in rapid etiological diagnosis.

Priming with FLO8-deficient Candida albicans induces Th1-biased protective immunity against lethal polymicrobial sepsis.

The morphological switch between yeast and hyphae of Candida albicans is essential for its interaction with the host defense system. However, the lack of understanding of host-pathogen interactions during C. albicans infection greatly hampers the development of effective immunotherapies. Here, we found that priming with the C. albicans FLO8-deficient (flo8) mutant, locked in yeast form, protected mice from subsequent lethal C. albicans infection. Deficiency of Dectin-2, a fungus-derived α-mannan recognition receptor, completely blocked flo8 mutant-induced protection. Mechanistically, the flo8 mutant-induced Dectin-2/CARD9-mediated IL-10 production in DCs and macrophages to block thymus atrophy by inhibiting the C. albicans-induced apoptosis of thymic T cells, which facilitated the continuous output of naive T cells from the thymus to the spleen. Continuous recruitment of naive T cells to the spleen enhanced Th1-biased antifungal immune responses. Consequently, depletion of CD4+ T cells or blockade of IL-10 receptor function using specific antibodies in mice completely blocked the protective effects of flo8 mutant priming against C. albicans infection. Moreover, mannans exposed on the surface of the flo8 mutant were responsible for eliciting protective immunity by inhibiting the C. albicans-induced apoptosis of thymic T cells to sustain the number of naive T cells in the spleen. Importantly, priming with the flo8 mutant extensively protected mice from polymicrobial infection caused by cecal ligation and puncture (CLP) by enhancing Th1-biased immune responses. Together, our findings imply that targeting FLO8 in C. albicans elicits protective immune responses against polymicrobial infections and that mannans extracted from the flo8 mutant are potential immunotherapeutic candidate(s) for controlling infectious diseases.

Epidemiological study of Trichosporon asahii infections over the past 23 years.

Trichosporon is a yeast-like basidiomycete, a conditional pathogenic fungus that is rare in the clinic but often causes fatal infections in immunocompromised individuals. Trichosporon asahii is the most common pathogenic fungus in this genus and the occurrence of infections has dramatically increased in recent years. Here, we report a systematic literature review detailing 140 cases of T. asahii infection reported during the past 23 years. Statistical analysis shows that T. asahii infections were most frequently reported within immunodeficient or immunocompromised patients commonly with blood diseases. Antibiotic use, invasive medical equipment and chemotherapy were the leading risk factors for acquiring infection. In vitro susceptibility, clinical information and prognosis analysis showed that voriconazole is the primary drug of choice in the treatment of T. asahii infection. Combination treatment with voriconazole and amphotericin B did not show superiority over either drug alone. Finally, we found that the types of infections prevalent in China are significantly different from those in other countries. These results provide detailed information and relevant clinical treatment strategies for the diagnosis and treatment of T. asahii infection.

Choosing the Right Antifungal Agent in ICU Patients.

Fungi are responsible for around 20% of microbiologically documented infections in intensive care units (ICU). In the last decade, the incidence of invasive fungal infections (IFI), including candidemia, has increased steadily because of increased numbers of both immunocompromised and ICU patients. To improve the outcomes of patients with IFI, intensivists need to be aware of the inherent challenges. This narrative review summarizes the features of routinely used treatments directed against IFI in non-neutropenic ICU patients, which include three classes of antifungals: polyenes, azoles, and echinocandins. ICU patients' pathophysiological changes are responsible for deep changes in the pharmacokinetics of antifungals. Moreover, drug interactions affect the response to antifungal treatments. Consequently, appropriate antifungal dosage is a challenge under these special conditions. Dosages should be based on renal and liver function, and serum concentrations should be monitored. This review summarizes recent guidelines, focusing on bedside management.

MICROMYCETES INFECTION IN PATIENTS WITH THERMAL TRAUMA.

GOAL: Determination of basic epidemiological parameters of burn patients with micromycetes infection. Identification of the most important micromycetes in burn patients. MATERIAL AND METHODS: Monocentre retrospective study enrolling all adult burn patients who were hospitalized between 2007 and 2015 and in whom micromycetes were isolated during hospitalization. ABSI index (Abbreviated Burn Severity Index) was used to evaluate severity of thermal trauma. Results were statistically analysed. RESULTS: There were 61 patients with thermal trauma identified in total during the period of observation, and there were yeast or fibrous fungi isolated. There were 37 males and 24 females (M:F ratio - 1.5:1) in this group. The average age of patients was 57.3 years (29 patients were aged up to 60 years, 32 patients were over the age of 60 years, inclusive). 6 patients died (lethality was 9.8%). The average extent of the burn area was 21.6% TBSA (median 14.0%). There were 90 strains of micromycetes cultured in total in these patients (79 yeasts, 11 fibrous fungi). Micromycetes were isolated from burn area in 30 patients, from the lower airways in 19 patients, from the urogenital area in 15 patients and from blood culture in 7 patients. Non-albicans Candida species were predominant among yeasts (60 strains); Candida albicans was isolated 16 times in total. Aspergillus fumigatus (4 isolations) and Fusarium species (2 isolations) were predominant species among fibrous fungi. CONCLUSION: We successfully identified the basic epidemiological parameters in burn patients with micromycetes infection, similarly to the most important yeasts and fibrous fungi causing infection in these patients.

[Consequences of autopsies for the living : Causes of death in the clinical diagnosis "septic and toxic shock"]. / Konsequenzen von Autopsiebefunden für die Lebenden : Todesursachenfeststellungen bei der klinischen Diagnose "septisch-toxischer Schock".

BACKGROUND: There is reason to believe that the diagnosis of septic and toxic shock, as indicated on the death certificate, cannot be confirmed as the cause of death without autopsy and subsequent histological analysis. The external examination of the corpse can therefore not represent the sole basis for a reliable statement about the infection status of a corpse, e. g. as a prerequisite for embalming. MATERIAL AND METHODS: The validity of autopsy in determining septic and toxic shock as the cause of death is demonstrated in 7 exemplary cases. RESULTS: Decades of experience in a university pathology institute have shown that an external examination of the corpse alone is not suitable for certifying the cause of death if an infectious disease is suspected. Consequently, only autopsy with subsequent histological analysis provides reliable statements on the etiopathogenesis of the underlying process. Possible problems and discrepancies between clinical and pathological diagnoses are discussed on the basis of several cases with or without autoptic confirmation of the septic shock. The case of a missionary from Africa infected with Lassa virus serves to point out the seriousness of the threat an undiagnosed infection may represent to the attending staff. CONCLUSION: During the treatment of patients suspected to have an infectious cause of fever of unknown origin, compliance with the usual safety regulations, including adequate disinfecting measures, is essential. In cases with fatal outcome, not infrequently under the clinical picture of a septic and toxic shock, autopsy should be regularly performed to confirm the type of infection and the infectious cause of death. Rapid and open communication between the professional groups involved plays a crucial role in this process.

Diffuse interstitial and multiple cavitary lung lesions due to Talaromyces marneffei infection in a non-HIV patient.

A 57-year-old man presented with unproductive cough and dyspnea for 6 months in Fujian Province, China. His misuse of a large amount of steroids (accumulated dose equivalent to 3530 mg prednisolone) resulted in Talaromyces marneffei infection. Chest computed tomographic scan revealed diffuse interstitial and multiple cavitary lung lesions. Treatment with amphotericin B combined with itraconazole resulted in total recovery, with marked regression of lung lesions.

Compare the nursing effects of 3 kinds of collutory for patients with chronic obstructive pulmonary diseases when combined with oral fungal infection / 中国实用护理杂志

Objective To compare the efficacy of three solutions used in the patients who suffered chronic obstructive pulmonary diseases (COPD ) when combined with oral fungal infection,and to find the most suitable one. Methods Using randomized controlled trial method, 98 patients who suffered COPD complicated with fungal infection were randomly divided into three groups. Method 1, using 0.5% povidone for the oral care. Method 2, making 200 million U of nystatin powder dissolved in 2.5% NaHCO3 for the oral care.Method 3, gave oral care with 2.5% NaHCO3 first, then put 200 million U of nystatin powder into 20 ml glycerol and covered the oral spots with them. Three groups were observed in the effect of oral fungal infection and the feedback from patients and nurses. Results In the analysis of the white spots sharply disappeared or pharyngeal swab smear turned negative, method 1 and method 3 had no statistical difference, P>0.05, but method 2 and method 3 had statistical significance, t=3.892, P 0.05. The time made solution of method 1 occupied 41.7% of method 3, and the time made solution of method 2 occupied 54.4%of method 3, while there was no statistical significance in total time of three groups (P>0.05). Conclusions The oral care with 0.5% povidone had good effect on the patients who suffered COPD complicated with oral fungal infection,with good comfort,and most nurses saving time. Nystatin powder dissolved in 2.5% NaHCO3 worked later than nystatin powder dissolved in glycerol.

Effects of immunocyte on the process of fungal keratitis / 中华实验眼科杂志

Background Infective keratopathy is a key cause of corneal blindness in China,and fungal keratitis is proved to have a higher incidence and bigger threats in infective keratitis.Researches showed that topical immunology plays an important effect during the development of fungal keratitis,but its mechanism is still studying.Objective This experiment was to explore the critical immunocyte during the process of fungal keratitis.Methods Forty-eight SPF 12-week-old male C57BL/6J mice were included and randomized into the control group and model group.The fungal keratitis model closely mimicking human cornea infections was established in the mouse using scratch followed by incubation of fusarium solani on the cornea,and the mice in the control group scratched on the cornea only.Cornea was examined under the slit lamp at 0,6,9,12,24,72 and 120 hours after operation.The severity of keratomycosis was clinically scored based on the literature criteria.The inflammatory cells were identified using immnofluorescence label,and the number of the inflammatory cells was calculated and compared among different groups and time points.This study complied with the Statement of ARVO in the use of experimental animal.Both Experimental Animal Ethic Commission in Zhengzhou University and Life Science Management Commission approved this study proposal.Results After inoculation of fusarium solani,typical fungul keratitis signs were seen on the cornea.Severe corneal opacifieation occurred within 24 hours and peaked at 72 hours.However,only mild edema of cornea was exhibited and gradually recovered normal in the control group within 24 hours.The clinical score of inflammation was higher in the model group in various time points than that in the control group,and it was seen that 24-72 hours after operation,the score attached peak in the model group with a significant difference in comparison with the control group(P＜0.01).In 9,12,24,72 and 120 hours after operation,the number of neutrophil cells was significantly increased in the model group compared with control group (P＜0.05),and that in 12,24,72 hours after operation was significantly higher than the 6 hours(P=0.004,0.000,0.001).However,no significant differences were seen in the number of neutrophil cells between 9 or 120 hours and 6 hours after operation(P=0.772,0.323).The number of T lymphocytes in cornea was significantly increased in 72 and 120 hours in comparison with 6 hours in the model group(P=0.000,0.000),and from 72 to 120 hours after operation,the number of T lymphocytes was significantly higher than that of the contral group (P＜0.01).The neutrophil cell number was positive correlated with the inflammatory score in the early phase (r =0.593,P =0.000).T limphocyte emerged in late phase but no significant correlation with the clinical score (r＝0.315,P=0.062).Conclusions Neutrophil cells play a critical role in the development of fungal keratitis in early stage.

Candidiasis in pediatric patients with cancer interned in a university hospital / Candidíases em pacientes pediátricos com câncer internados em um hospital universitário

Fungi are common causes of infection in immunocompromised patients. Candida species are frequently involved in these cases. In order to investigate candidiasis in pediatric patients with cancer, clinical samples were collected from one hundred and twenty two patients interned in the Oswaldo Cruz University Hospital in Recife, Brazil. Yeasts were isolated from thirty-four clinical samples. The species isolated were: Candida albicans (fourteen isolates), C. parapsilosis (nine isolates), C. guilliermondii (two isolates) and C. tropicalis (two isolates). We found that candidemia was most frequent in patients with malignant hematology and that C. parapsilosis infections caused the highest mortality.

Os fungos são causas comuns de infecções em pacientes imunocomprometidos e espécies de Candida são freqüentemente envolvidas nesses casos. A fim de investigar infecção fúngica em pacientes pediátricos com câncer, amostras clínicas foram coletadas de cento e vinte dois pacientes internados no Hospital Universitário Oswaldo Cruz em Recife, Brasil. Leveduras foram isoladas de trinta e quatro amostras clínicas. As leveduras isoladas foram: Candida albicans (catorze isolados), C. parapsilosis (nove isolados), C. guilliermondii (dois isolados) e C. tropicalis (dois isolados). Descobrimos que candidemia foi mais freqüente em doentes com hematologias malignas e que C. parapsilosis apresentou maior mortalidade.

Candidiasis in pediatric patients with cancer interned in a university hospital.

Fungi are common causes of infection in immunocompromised patients. Candida species are frequently involved in these cases. In order to investigate candidiasis in pediatric patients with cancer, clinical samples were collected from one hundred and twenty two patients interned in the Oswaldo Cruz University Hospital in Recife, Brazil. Yeasts were isolated from thirty-four clinical samples. The species isolated were: Candida albicans (fourteen isolates), C. parapsilosis (nine isolates), C. guilliermondii (two isolates) and C. tropicalis (two isolates). We found that candidemia was most frequent in patients with malignant hematology and that C. parapsilosis infections caused the highest mortality.