A Case Report of Mucormycosis in a Patient with a Recent History of COVID-19 Infection in India

Introduction: Mucormycosis is an angio-invasive fungal infection that increased significantly during the 2nd wave of the Covid-19 pandemic in India. The rise of cases was attributed to inflammatory changes, poor quality oxygen, immune suppression, and corticosteroid therapy. Case Presentation: This case study reports the history, treatment, and rehabilitation of a case of post-Covid-19 mucormycosis infection. The patient was admitted to the hospital following respiratory distress, at the beginning of the 2nd Covid-19 wave in India. Intravenous antibiotics, steroids, and moist O2 were administered, intensive support was provided and the patient was discharged after 13 days. Following extraction of 17, the patient reported signs of oro-antral communication which was managed by performing antral lavage and buccal advanced flap closure. Histopathological investigation of tissue salvaged during the procedure revealed the presence of fungal hyphae. Management and Prognosis: Following diagnosis, anti-fungal medication was prescribed, and a maxillectomy was performed to remove the affected tissue. On follow-up, the tissue healed with no further complications or symptoms, and rehabilitation was performed using an obturators and are movable complete denture. Histopathological investigations were carried out on the tissue salvaged during maxillectomy which confirmed Mucormycosis infection. Conclusion: The importance of histopathological investigation in the diagnosis of any infectious disease is enumerated in this paper

Infection of perennial ryegrass (Lolium perenne) by an endophyte fungus (Neotyphodium lolii) decreases the abundance and diversity of predators and parasitoids

ABSTRACT Perennial ryegrass is one of the most important food sources in animal production. However, several pests affect this crop, and one of the primary control strategies is the symbiotic relationships between ryegrass endophyte fungi. This fungus produces alkaloids that exhibit toxic activity against arthropods. Furthermore, the effect of fungi may extend to higher trophic levels, including predators (spiders and/or insects), decreasing their abundance and diversity. Given the importance of spiders and insects as predators, whether the symbiotic interaction between perennial ryegrass and endophyte fungus reduces the abundance and diversity of predators pose an important question. To address this question, natural enemies in perennial ryegrass were collected and analyzed over a year, and the percentage of endophyte fungus was evaluated by the presence of hyphae from two ryegrass cultivars, Jumbo (E-) and Alto AR1 (E+). We observed an 80% endophyte infection rate for (E+) and 0% for (E-). Moreover, 222 individual spiders corresponding to 10 families were identified in both perennial ryegrasses, including 209 individuals for (E-) and 13 for (E+). The most abundant spider family was Lycosidae, representing 71.17% of the total spiders. In addition, 65 insects were collected, corresponding to 6 families, with Carabidae being the most abundant. Furthermore, the Simpson index indicated the dominance of the family Lycosidae. Overall, spider and insect abundance and diversity were reduced in (E+), suggesting a negative effect of the endophyte on predator populations.

Investigation on the antifungal activity of pyranium derivatives N2 / 药学实践杂志

Objective To study the antifungal activity of N2 derivatives. Methods The anti-fungal activity of N2 compounds was investigated by micro-liquid dilution. Then the activity of N2 compounds on hyphal and biofilm formation was investigated. Results N2 compounds had significant antifungal activity against Candida albicans. It also expressed actively inhibitory effect on hyphal and biofilm formation. The mechanism of its fungicidal function was to damage the structure of candida albicans’ cell membrane and cell wall. Conclusion The results showed that N2 had obvious antifungal activity against Candida albicans., which provided a new idea for the development of antifungal drugs and the solution of antifungal drugs resistance.

Structure and mechanism of Candida albicans Int1 involved in septin organization regulation / 中华微生物学和免疫学杂志

Objective:To investigate the mechanism of Candida albicans Int1 in regulating septin organization. Methods:A series of full-length and truncated fragments of Int1 were constructed and fused with green fluorescent protein (GFP). The intracellular localization of the fusion proteins was observed under a fluorescence microscope. The region in Int1 that was required for bud neck localization was identified. Full-length and fragments of Int1 were overexpressed in the yeast Saccharomyces cerevisiae and the changes in cell growth, cell morphology and septin organization were investigated to determine the functional region in Int1 that mediated the interaction with septin. Moreover, the co-localization of the region and septin was analyzed. Results:The full-length Int1 consisted of 1 661 amino acid residues. A middle region of 209 amino acid residues, Int1-M4 (739-947 aa), that could be localized at the bud neck during both small and large bud periods was identified. Overexpression of Int1-M4 led to significant growth defects, elongated bud and disorganized septin. In the cells with elongated bud, Int1-M4 and septin with abnormal structures could be co-localized.Conclusions:Int1-M4 (739-947 aa), the middle region of Int1 containing 209 amino acid residues, mediated the bud neck localization and the interaction with septin, playing an important role in regulating septin organization.

Mucormicosis Maxilar posterior, secundaria a una exodoncia: Reporte de un caso y revisión de la literatura

RESUMEN: La Mucormicosis se describe como una infección fúngica de tipo oportunista y potencialmente mortal, reportándose en la literatura como la tercera en frecuencia, muy por detrás de la Candidiasis y Aspergilosis. Esta infección es causada por hongos de la familia de los Mucorales. Se presenta el caso de un hombre con Diabetes Mellitus II con un control metabólico deficiente, el cual posterior a una exodoncia, presenta una Mucormicosis Maxilar, presentando una etiología atípica y logrando ser confirmada posterior a biopsias y pruebas de cultivo, demostrando elementos del tipo Mucor. Se realizó una Maxilectomía parcial del lado afectado como tratamiento con una evolución favorable. Esta revisión destaca la importancia de la búsqueda activa basada en la semiología y la importancia de los exámenes complementarios, implicando lograr un correcto diagnóstico y pronóstico de la enfermedad.

ABSTRACT: Mucormycosis is described as an opportunistic and life-threatening fungal infection, being reported in the literature as the third in frequency, far behind candidiasis and aspergillosis. This infection is caused by fungi of the Mucorales family. We present the case of a man with Diabetes Mellitus II with poor metabolic control who, after an extraction, presents a Maxillary Mucormycosis, with an atypical etiology and confirmed after biopsies and culture tests, demonstrating elements of the Mucor type. A partial maxillectomy was performed on the affected side as a treatment with a favorable outcome. This review highlights the importance of active search based on semiology and of complementary examinations, implying a correct diagnosis and prognosis of the disease.

Neutrophils phagocytosing fungal hyphae in urinary sediment / Neutrófilos fagocitando hifas no sedimento urinário

Abstract The Phagocytosis of fungal structures by neutrophils is a well-documented function of these immune cells. However, neutrophil phagocytosis of hyphal structures in the urine sediment is not usually observed during routine sample evaluation. This is a case of hyphal phagocytosis by neutrophils in the urine of a kidney allograft recipient patient.

Resumo A fagocitose de estruturas fúngicas por neutrófilos é uma função bem documentada destas células imunes. No entanto, a fagocitose de hifas por neutrófilos no sedimento urinário não é normalmente observada durante avaliação de rotina de amostras. Este é um caso de fagocitose de hifas por neutrófilos na urina de um paciente receptor de aloenxerto renal.

Study on the antifungal activity of phenazine derivatives / 药学实践杂志

Objective To study the antifungal activity of phenazines derivatives. Methods The anti-fungal activity of phenazine compounds was evaluated initially with micro-liquid dilution. No significant antifungal activity against Candida albicans was found. Then, with the combination of phenazine compounds and fluconazole, the anti-fungal activity against fluconazole-resistant C. albicans was detected. Results The phenazine-17 had significant antifungal activity when combined with fluconazole through the inhibition of hyphae formation. Conclusion This study provides a new idea for the development of antifungal drugs and the solution of antifungal drug resistance.

Hyphal Formation is Enhanced by 121°C Heat Inactivation of Serum Added to Culture Media

Background: A novel method for inducing hyphae formation, was described.Methods:Mycelia of experimental and clinical strains from 100 patients with oral lichen planuswere cultured in RPMI 1640 medium with 56°C or 121°C inactivated calf sera. After these strains were cultured for 2 h to 7 days, the average hyphae formation rate was observed under light microscopy. Results: The 121°C inactivated calf serum showed better outcomes than 56°C inactivated calf serum in terms of time and amounts of hyphae products. Conclusions:For Candida albicanshyphae culture, the 121°C inactivated serum is superior to the 56°C inactivated serum

Study on the antibiofilm activity of kaempferol in Candida albicans / 药学实践杂志

Objective To study the action of kaempferol (KAE) against Candida albicans biofilms and explore the potential mechanisms. Methods Biofilm metabolic activity assay was used to investigate the action of KAE against C. albicans biofilm formation as well as mature biofilm. The inhibition of KAE in hyphal formation was examined by microscope. The water-hydrocarbon two-phase separation assay was used to test the effect of KAE on the cell surface hydrophobicity of C. albicans. The mRNA expression of the genes involved in biofilm formation was determined by real time RT-PCR. Results KAE showed inhibition effect on C. albicans biofilm formation in a dose-dependent manner. Moreover, KAE inhibited mature biofilm. The biomass of biofilm was reduced upon KAE treatment. KAE inhibited hyphal formation and reduced the cell surface hydrophobicity of C. albicans. In the presence of KAE, the mRNA expression of the genes involved in biofilm formation was changed, with the up-regulation of BCR1,NRG1,TUP1 and down-regulation of HWP1,EFG1,CPH1,ALS1,ALS3 and CSH1. Conclusion KAE showed antifungal activity against C. albicans biofilm. The mechanisms may relate to the inhibition of hyphal formation and reduction of cell surface hydrophobicity.

Esophageal pythiosis in an ostrich (Struthio camelus) / Ptiose Esofágica em avestrus (Struthio camelus)

Uma avestruz-do-pescoço-vermelho, com dois anos de idade, apresentava um nódulo no terço médio do esôfago e foi submetida a procedimento cirúrgico. Histologicamente, observou-se uma área focalmente extensa de necrose estendendo-se da túnica mucosa à muscular, e, em algumas secções, à túnica adventícia. Circundando a área de necrose, observou-se uma reação inflamatória composta principalmente por granulócitos e macrófagos, associada à fibroplasia e neovascularização. Em meio às áreas de necrose e inflamação, verificavam-se numerosas imagens negativas de hifas em seções longitudinais e transversais, melhor apreciadas pela coloração de metenamina nitrato de prata de Grocott. O diagnóstico definitivo de infecção por Pythium insidiosum foi confirmado por imuno-histoquímica. A avestruz recebia água para consumo de um lago localizado em uma área de pastagem, no qual alguns cavalos haviam desenvolvido pitiose cutânea anteriormente.(AU)

Presence of pathogenic candidal hyphae in patients with palatal coverage appliances/prostheses

@#Oral candidiasis is an opportunistic infection of the oral cavity. This study aimed to determine prevalence of oral candidiasis in patients with palatal coverage orthodontic appliances and prostheses and its risk factors. Three groups of patients were recruited after an informed consent. The two tested groups were the denture and the upper removable orthodontic appliance (URA) groups while patients with no prosthesis/appliance acted as control. The assessments included demographic profile, denture/URA age, night-time wearing and duration of wearing. Signs and symptoms of candidiasis were examined intra-orally. Unstimulated salivary flow rate test was also performed. Smears samples were taken from palatal mucosa and impression surfaces of denture/URA and stained with Periodic Acid Schiff (PAS). The data were analysed using descriptive and chisquare tests. A total of 86 patients were recruited; denture (n=30), URA (n=22), control (n=34). Denture and URA groups had significantly more positive cases of candidal infection (56.7 and 72.7% respectively) as compared to control group (20.6%). Among the subjects with oral candidiasis, denture patients were detected to have more severe amount of candidal hyphae (20%) than URA (13.6%) group. The relationships between oral candidiasis and these risk factors i.e. age, night-time wearing, daily total hours of wearing and denture/URA hygiene were statistically significant (p<0.05). Thus, patients who are wearing palatal coverage removable appliances or prostheses have higher prevalence of oral candidiasis. The risk is greater with confounding factors such as age, duration of wearing and oral hygiene

Changes in transcriptome expression profiles and functional analysis of differentially expressed genes during mycelium-to-yeast transformation of Sporothrix schenckii / 中华皮肤科杂志

Objective To compare and analyze the differences in the transcriptomics between mycelium and early yeast phases of Sporothrix schenckii (S.schenckii),and to realize the changes in transcriptome expression profiles during mycelium-to-yeast transformation.Methods A standard strain of S.schenckii (ATCC 10268) was subjected to 96-hour culture with Sabouraud medium at 25 ℃ or 36-hour culture with brain-heart infusion medium at 37 ℃ to obtain the mycelium and yeast form of S.schenckii,and then,their transcriptomes were sequenced.Functional annotation was performed for screened unigenes by comparison using several databases (such as NR,Swiss-Prot,KEGG,COG,KOG,GO and Pfam),coding sequence prediction,and gene expression analysis in each sample.Finally,the differentially expressed genes were subjected to pattern clustering,functional annotation and enrichment analysis.Results A total of 14.76 Gb valid data (clean data) were obtained,and functional annotation results were acquired in 28 094 of 43 863 assembled unigene clusters.Compared with S.schenckii in mycelium phase,there were 10 969 up-regulated genes and 199 down-regulated genes in S.schenckii in yeast phase.These differentially expressed genes were involved in protein phosphorylation,intracellular protein transport,cellular protein modification,small guanosine triphosphate-mediated signal transduction,vesicle-mediated transport,translation,intracellular signal transduction,microtubule formation,adenosine triphosphate synthesis,coupled proton transport and so on.Sixteen genes in the mitogen-activated protein kinase (MAPK) pathway and two-component signaling pathway,which were two important signal transduction pathways involved in fungal morphogenesis,and 16 genes involved in chitin synthesis and metabolism were all confirmed to be up-regulated in S.schenckii in yeast phase.Conclusions Compared with S.schenckii in mycelium phase,great changes in gene expression profiles were observed in S.schenckii in yeast phase.These differentially expressed genes are involved in many functions,suggesting that the dimorphic transition of S.schenckii is regulated by a multi-gene network system.

Inhibitory effect of extract of Coptidis Rhizoma on invasion of Candida albicans hyphae in vitro / 中国中药杂志

The aim of this paper was to investigate the inhibitory effect of extract of Coptidis Rhizoma(ECR) on invasion of Candida albicans hyphae in vitro.XTT reduction method was used to evaluate the metabolic activity of C.albicans.The colony edge growth of C.albicans was observed by solid medium.The growth of C.albicans hyphae was determined on semi-solid medium.The morphology and viability changes of C.albicans hyphae were assessed by scanning electron microscope and fluorescence microscope.qRT-PCR method was used to detect the ALS3 and SSA1 expression of C.albicans invasin genes.The results showed that the metabolic viability by XTT method detected that the activity of C.albicans was gradually decreased under the intervention of 64,128 and 256 mg·L-1 of ECR respectively.128,256 mg·L-1 of ECR significantly inhibited colony folds and wrinkles on solid medium and the hyphal invasion in semi-solid medium.Scanning electron microscopy and fluorescence microscopy showed that 128,256 mg·L-1 of ECR could inhibit the formation of C.albicans hyphae.qRT-PCR results showed that the expression of invasin gene ALS3 and SSA1 was down-regulated,and especially 256 mg·L-1 of ECR could down-regulate the two genes expression by 4.8,1.68 times respectively.This study showed that ECR can affect the invasiveness of C.albicans by inhibiting the growth of hyphae and the expression of invasin.

Dynamic distribution of calmodulin labeled with red fluorescent protein in the polar growth of Aspergillus fumigatus / 中华皮肤科杂志

Objective To construct a native promoter-regulated Aspergillus fumigatus strain containing red fluorescent protein-labeled calmodulin (CaM-RFP),and to observe the dynamic distribution of calmodulin during the growth of Aspergillus fumigatus.Methods Bilateral flanking sequences of Aspergillus fumigatus calmodulin gene were designed,and plasmids containing the two flanking sequences and mRFP-Aspergillus fumigatus pyrG gene (mRFP-AfpyrG) were amplified separately.The final linear PCR product for transformation was generated from the above three PCR products by fusion PCR.Then,the above linear fragment was transferred into the Aspergillus fumigatus strain by protoplast transformation,so as to construct the CaM-RFP Aspergillus fumigatus strain.The monoclonal colony was picked from the screening medium and subjected to culture.Then,the stablest fluorescent monoxenic strain of Aspergillus fumigatus was selected,and the transformant was verified by PCR.The recombinant strain and wild-type stain were cultured on solid nutrient media separately,and the morphology of these strains was observed by fluorescence microscopy at different time points.Additionally,the above 2 strains were cultured in liquid media separately,and XTT assay was performed to evaluate the growth activity of strains.Microscopy was also conducted to dynamically observe the CaM-RFP Aspergillusfumigatus strain,and analyze the spatial and temporal distribution of calmodulin during the growth and development of Aspergillus fumigatus.Results The fluorescent phenotype and PCR identification results both indicated the successful construction of the CaM-RFP Aspergillus fumigatus strain.The growth activity at 24 hours did not differ between the recombinant strain and wild-type stain (A490:0.689 ± 0.081 vs.0.678 ± 0.054,t =1.32,P ＞0.05),so did the morphology.During the polarized growth of Aspergillus fumigatus,calmodulin was always at the top of the hyphae,germination site of the hyphal branch and the top of new branches.Conclusion Calmodulin may be involved in the regulation of spore germination and polar hyphal growth of Aspergillus fumigatus.

Death of pastures syndrome: tissue changes in Urochloa hybrida cv. Mulato II and Urochloa brizantha cv. Marandu / Síndrome da morte das pastagens: Alterações teciduais em Urochloa hybrida cv. Mulato II e Urochloa brizantha cv. Marandu

Abstract The quality of forage production is a prerequisite to raising livestock. Therefore, income losses in this activity, primarily cattle raising, can result in the impossibility of economic activity. Through the qualitative and quantitative anatomical study of Urochloa hybrida cv. Mulato II and U. brizantha cv. Marandu, we searched for descriptions and compared changes in the individual vegetative body from populations with death syndrome pastures (DPS). Specimens were collected at different physiological stages from farms in northern Mato Grosso. After collection, the individuals were fixed in FAA50 and stored in 70% alcohol. Histological slides were prepared from the middle third of the sections of roots, rhizomes, and leaves, and the proportions and characteristics of tissues were evaluated in healthy, intermediate, and advanced stages of DPS. Changes were compared between cultivars. With the advancement of the syndrome, the following changes were observed: a more marked decrease in the length of roots in U. hybrida; disorganization of the cortical region of the roots and rhizome cultivars; fungal hyphae in roots and aerenchyma formation in U. hybrida; a decrease in sclerenchyma fiber proportions in roots and leaves; sclerification of the epidermis of U. brizantha rhizomes; and an increase in pericyclic fibers in U. hybrida. Furthermore, there was a decrease in the volume of epidermal cells of the abaxial face of the leaves of both cultivars, with a greater reduction in U. hybrida; a gradual decrease in thickness in the midrib of leaves similar to leaf mesophyll; conduction system obstructions; partial or total cell lysis in roots and rhizomes affected by the syndrome. Obstructions in sieve tube element and companion cells, and sometimes obstruction in xylem vessel elements. The evolution of DPS in cultivars was similar, but there were variations, arising probably from the physiological response to stress, such as aerenchyma formation in the root and increased pericycle in the rhizome of U. hybrida.

Resumo A produção de forragem de qualidade é um pressuposto indispensável à atividade pecuária moderna. Dessa forma, perdas de rendimento nesta atividade preliminar à criação bovina, podem inviabilizar esta atividade econômica. Através de estudos anatômicos qualitativos e quantitativos em Urochloa hybrida cv. Mulato II e U. brizantha cv. Marandu buscou-se descrever e comparar as alterações do corpo vegetativo de indivíduos de populações que apresentam síndrome da morte das pastagens (SMP). Foram coletados indivíduos em estádios morfológicos distintos em propriedades rurais no norte de Mato Grosso, Amazônia Meridional. Após a coleta, os indivíduos foram fixados em FAA50 e armazenados em álcool 70%. Lâminas histológicas foram confeccionadas a partir de secções do terço médio de raízes, rizomas e folhas, onde foram avaliadas as características e proporções dos tecidos em indivíduos saudáveis, indivíduos em estádio intermediário e também em estádio mais avançado da SMP, bem como comparadas as alterações entre as duas cultivares. Com o avanço da síndrome verificou-se diminuição mais acentuada da extensão de raízes em U. hybrida. A região cortical das raízes e do rizoma das cultivares exibiu desorganização, sendo observada nas raízes a presença de hifas fúngicas e a formação de aerênquima em U. hybrida. As proporções de fibras esclerenquimáticas diminuíram nas raízes e folhas. No rizoma, a epiderme de U. brizantha apresentou esclerificação e U. hybrida exibiu aumento da quantidade de fibras pericíclicas. Houve diminuição do volume das células epidérmicas da face abaxial das folhas de ambas as cultivares, com maior diminuição em U. hybrida. A região da nervura central das folhas apresentou gradativa diminuição com espessura similar ao mesofilo foliar. O sistema de condução apresentou obstruções, onde elementos de tubo crivado e células companheiras do floema das raízes e rizomas afetados pela síndrome exibiram parcial ou total lise celular e os elementos de vaso do xilema por vezes se mostraram obstruídos. A evolução da SMP nas cultivares estudadas é semelhante, contudo houve variações decorrentes provavelmente da resposta fisiológica ao estresse, como a formação de aerênquima na raiz, alteração e aumento da espessura do periciclo do rizoma de U. hybrida.

Inhibition Mechanism of Rohdea Rothon hypha formation by Candida albicans / 中国生化药物杂志

Objective To investigate the inhibition mechanism of rohdea roth on hyphae formation by Candida albicans.Methods MTT assay was used to detect the minimum inhibitory concentration(MIC)and minimum fungicidal concentration(MFC)of C.albicans.The inhibitory effect of fungicidal adherence was detected by MTT assay.Inverted fluorescence microscope was used to observe effect on hyphae formation.The influence on Efg1 and Hwp1gene expression were detected by RT-PCR method.ResultsMIC and MFC of C.albicans were 16 mg/mL and 32 mg/mL, respectively.The inhibitory effects of rohdea roth on C.albicans adherence and hyphae formation were significantly inhibited,and the concentration was dose-dependent.After the concentration of 16 mg/mL acted on C.albicans for 6 h, hyphae disappeared completely.The results of RT-PCR showed that the gene expression of Efg1 and Hwp1 could be inhibited by rohdea roth.Compared with the control group,the expression of Efg1 and Hwp1in the experimental groupwere reduced by 84.18% and 59.57%(P<0.01).Conclusion The inhibitory effect of rohdea roth on the adherence and hyphae formation of C.albicans is mainly through inhibiting the expression of Efg1 and Hwp1genes.

Study of the detection and significance of intracellular bacterial communities in patient with catheter-associated urinary tract infections / 中华泌尿外科杂志

Objective To detection the urine of bacteria hyphae and intracellular bacterial communities in patients with indwelling urinary catheter and discuss intracellular bacterial comnmunities in the pathogenesis of catheter-related urinary tract infection.Methods From May 2014 to February 2016,95 cases with D-J stent indwelling were enrolled in this study,including 38 male patients and 57 female patients.The mean age was (43 ±21)years old,ranging from 25 to 83 years old.We recorded those patient g clinical symptoms,middle urine culture results.If the middle urine culture was positive,further pathology test and scanning electron microscopy for bacteria hyphae and intracellular bacterial communities would be considered.Results The middle urine culture showed positive in 21 cases (22％,21/95);The classification of bacteria included E.coli in 11 cases,dung enterococcus in 2 cases,klebsiella pneumonia in 4 cases,pseudomonas aeruginosa in 3 cases,epidermis staphylococcus aureus in 1 case.Among those 21 patients,9 cases had the symptoms of fever and shiver.Urine pathology testing found hyphae in 6 cases (6％,6/95).all others were E.coli infection.For scanning electron microscope,6 cases were found rodshaped bacteria and hyphae.3 cases were found intracellular bacterial communities.Conclusions The presence of intracellular bacterial communities made urothelial itself the source of endogenous bacteria of urinary tract infection.Catheter-related urinary tract infections in patients with recurrence maybe basically homology bacteria.

Fator de necrose tumoral alfa TNF-α inibe a formação do biofilme de Candida albicans

O presente estudo teve como objetivo investigar a ação do TNF-α frente ao biofilme deCandida albicans. Para tanto, cepas de C. albicans fracamente produtoras de biofilme(49BMv e 51BMv) e fortemente produtoras de biofilme (ATCC 10231 e CEMM 01.05.006)foram incubadas em meio RPMI em placas de 96 poços, a 35 ºC por 48 ou 96 horas paraavaliar o desenvolvimento do biofilme em formação e maduro, respectivamente, na ausênciaou presença de diferentes concentrações de TNF-α (0,1, 0,5, 1, 10, 20 e 40 ng/mL). A adiçãodo TNF-α foi realizada no momento do preparo do cultivo para avaliar seu efeito no biofilmeem formação ou após 48 horas de cultura para avaliação no biofilme maduro. Apósincubação, a atividade metabólica foi avaliada por meio do ensaio de redução de XTT. Aconcentração inibitória mínima (CIM) do TNF-α em C. albicans foi avaliada, dentre asconcentrações testadas 0,5, 1, 10 e 20 ng/mL, por meio da técnica de microdiluição em caldo,segundo documento M27-A3, preconizada pelo CLSI. Foi realizada uma análise da relaçãohifas/blastoconídios do biofilme por microscopia óptica, por meio de uma contagem dasformas filamentosas (hifas e pseudo-hifas) e leveduriformes e em seguida aplicada umafórmula que permitiu visualizar a relação entre a quantidade de formas filamentosas e asdemais estruturas. Os resultados indicaram uma marcante inibição da formação do biofilme deforma dose-dependente e uma discreta inibição do biofilme maduro pelo TNF-α...

The aim of this study was to investigate the activity of TNF-α against Candida albicansbiofilm. Therefore, week biofilm producer (49BMv and 51BMv) and strong biofilm producer(ATCC 10231 and CEMM 01.05.006) strains of C. albicans were incubated in RPMI mediumin 96-well plates at 35 °C for 48 or 96 hours in order to assess biofilm formation and maturebiofilm, respectively, in the absence or presence of different concentrations of TNF-α (0.1,0.5, 1, 10, 20 and 40 ng/ml). TNF-α was added during preparation of the biofilm to assess itseffect on biofilm formation or after 48 hours of culture for evaluation in mature biofilm. Afterincubation, the metabolic activity was assessed using XTT reduction assay. The minimuminhibitory concentration (MIC) of TNF-α against C. albicans was evaluated by brothmicrodilution technique according to Document M27-A3, recommended by CLSI, using theconcentrations 0.5, 1, 10 and 20 ng/mL. An analysis of the hyphae/blastoconidia ratio inbiofilm was performed using optical microscopy by couting of filamentous (hyphae andpseudo-hyphae) and yeast cells and then a formula which allowed visualization of therelationship between the amount of filamentous and other structures was applied. The resultsindicated that TNF-α showed a significant inhibition of biofilm formation in a dose-dependentway and a discrete mature biofilm inhibition. In addition, the analysis of hyphae/blastoconidiaratio showed a statistically significant reduction in the number of filamentous forms inbiofilm formation in the presence of TNF-α...

Candidiasis bucal en pacientes con diabetes mellitus / Oral Candidiasis in patients with diabetes mellitus

Se realizó un estudio descriptivo y transversal de 75 pacientes con diabetes mellitus, quienes presentaron candidiasis bucal, tratados en la consulta estomatológica del Policlínico de Especialidades del Hospital Provincial Docente Clinicoquirúrgico "Saturnino Lora Torres" de Santiago de Cuba, desde marzo de 2013 hasta igual mes de 2015, a fin de evaluar los resultados del diagnóstico clínico e histopatológico de la citada afección. Las muestras para biopsias se fijaron y procesaron con la técnica clásica de inclusión en parafina; se colorearon con una tinción especial de ácido peryódico de Schiff, lo cual confirmó la existencia de cambios celulares. Predominaron el sexo masculino, el grupo etario de 60 años y más; la ausencia de manifestaciones clínicas, el paladar duro y la comisura labial como los sitios de mayor prevalencia y la candidiasis eritematosa como la forma clínica más común. La presencia de hifas patógenas y células levaduriformes fueron los hallazgos histopatológicos más sobresalientes.

A descriptive and cross-sectional study of 75 patients with diabetes mellitus who presented oral candidiasis, treated in the stomatological service of the Specialties Polyclinic of "Saturnino Lora Torres" Teaching Provincial Clinical Surgical Hospital in Santiago de Cuba was carried out, from March, 2013 to the same month in 2015, in order to evaluate the results of the clinical and histopathologic diagnosis of the mentioned affection. The samples for biopsies were fixed and processed with the classic technique of inclusion in paraffin; they were colored with a special stain of Schiff periodic acid, which confirmed the existence of cellular changes. There was a prevalence of male sex, the 60 years and over age group; the absence of clinical symptoms, the hard palate and the corner of the mouth as the sites of greater prevalence and the erythematous candidiasis as the most common clinical form. The presence of pathogen hyphae and yeast-like cells were the most outstanding histopathologic findings.

Inhibitory effects of curcumin on hyphal development and biofilm formation of five kinds of non-Candida albicans / 中草药

Objective: To observe the inhibitory effects of curcumin on hyphal development and biofilm formation of five kinds of non-Candida albicans. Methods: Serial 2-fold dilution assay was used to determine the MICs of curcumin to C. tropicalis, C. glabrata, C. parapsilokis, C. krusei, C. guilliermondii; XTT assay was used to determine the SMIC50 of curcumin to the five non-C. albicans. Inverted microscope and scanning electron microscope (SEM) were applied to inspectting the morphological change of non-C. albicans treated by curcumin, The dilution method was applied to inspecting the hyphae around the colonies. Results: The MICs of curcumin to C. tropicalis, C. glabrata, C. parapsilokis, C. Krusei, C. guilliermondii were 64, 128, 256, 256, and 128 μg/mL, and the SMIC50 were 512, 512, >512, >512, and 512 μg/mL, respectively. We found that curcumin could inhibit the hyphal and biofilm formation of the five kinds of non-C. albicans by observation with inverted microscope and SEM, and curcumin could inhibit the hyphal development around the colonies by observation with optical microscope. Conclusion: Curcumin could inhibit the hyphal development and biofilm formation of the five kinds of non-C. albicans.