Tissue reaction to Aroeira (Myracrodruon urundeuva) extracts associated with microorganisms: an in vivo study.

Based on aroeira's (Myracrodruon urundeuva) antimicrobial activity and a future trend to compose intracanal medication, the aim of this study was to assess in vivo inflamatory tissue response to the extracts by edemogenic and histological analysis containing inactivated facultative and anaerobic microorganisms. For edema quantification, eighteen animals were divided into three groups (n = 3, periods: 3 and 6 hours) and 0.2 mL of 1% Evans blue per 100 g of body weight was injected into the penile vein under general anesthesia. After 30 min the animals received a subcutaneous injection in the dorsal region of aqueous or ethanolic extract of aroeira or saline (control) containing inactivated bacteria. Samples were collected, immersed in formamide for 72h, and evaluated by spectrophotometry (630 m). For histological analysis, polyethylene tubes with the extracts were implanted in the dorsal of 30 male rats. Analysis of the fibrous capsule and inflammatory infiltrate were performed after 7 and 30 days. The aqueous extract group induced less edema in both postoperative periods compared to the other groups, but the differences were not significant (p > 0.05). Tissue repair was significantly better after 30 days than after 7 days (p < 0.01). The aqueous solution showed less inflammatory response than the ethanolic solution (p < 0.05), with tendency for better results than control after 7 days. After 30 days, the response to both extracts was similar to control. The aqueous and ethanolic aroeira extracts containing inactivated microorganisms showed a trend for better results than saline, even when associated with microorganisms, and facilitated the tissue repair process.

Tissue reaction to Aroeira (Myracrodruon urundeuva) extracts associated with microorganisms: an in vivo study

Abstract: Based on aroeira's (Myracrodruon urundeuva) antimicrobial activity and a future trend to compose intracanal medication, the aim of this study was to assess in vivo inflamatory tissue response to the extracts by edemogenic and histological analysis containing inactivated facultative and anaerobic microorganisms. For edema quantification, eighteen animals were divided into three groups (n = 3, periods: 3 and 6 hours) and 0.2 mL of 1% Evans blue per 100 g of body weight was injected into the penile vein under general anesthesia. After 30 min the animals received a subcutaneous injection in the dorsal region of aqueous or ethanolic extract of aroeira or saline (control) containing inactivated bacteria. Samples were collected, immersed in formamide for 72h, and evaluated by spectrophotometry (630 m). For histological analysis, polyethylene tubes with the extracts were implanted in the dorsal of 30 male rats. Analysis of the fibrous capsule and inflammatory infiltrate were performed after 7 and 30 days. The aqueous extract group induced less edema in both postoperative periods compared to the other groups, but the differences were not significant (p > 0.05). Tissue repair was significantly better after 30 days than after 7 days (p < 0.01). The aqueous solution showed less inflammatory response than the ethanolic solution (p < 0.05), with tendency for better results than control after 7 days. After 30 days, the response to both extracts was similar to control. The aqueous and ethanolic aroeira extracts containing inactivated microorganisms showed a trend for better results than saline, even when associated with microorganisms, and facilitated the tissue repair process.

Importance of multidisciplinary approach in diagnosis of histoplasmosis.

A patient with systemic involvement, initially treated as tuberculosis, is presented in this report. There were only two painful subcutaneous nodules, from which we arrived at the correct diagnosis of histoplasmosis. The patient was attended by several experts in the fields of infectious diseases, nephrology and internal medicine, but the diagnosis was only possible after dermatological examination and skin biopsy. This case values multidisciplinary interaction between dermatologists and other medical areas for diagnosis of cases with atypical manifestations.

Importance of multidisciplinary approach in diagnosis of histoplasmosis

Abstract: A patient with systemic involvement, initially treated as tuberculosis, is presented in this report. There were only two painful subcutaneous nodules, from which we arrived at the correct diagnosis of histoplasmosis. The patient was attended by several experts in the fields of infectious diseases, nephrology and internal medicine, but the diagnosis was only possible after dermatological examination and skin biopsy. This case values multidisciplinary interaction between dermatologists and other medical areas for diagnosis of cases with atypical manifestations.

Phaeohyphomycosis Caused by a Novel Species, Pseudochaetosphaeronema martinelli.

Among the opportunistic mycoses that are emerging in patients with immunosuppression or severe underlying illness, many isolates lack of characteristic sporulation and until recently could not be identified. Clinical signs are mostly nonspecific and therefore such infections have often been disregarded. In the present paper we describe a novel, nonsporulating fungal species causing subcutaneous phaeohyphomycosis in two patients of different origin. One is a 73-year-old female from Martinique who suffered from rheumatoid arthritis, while the other case concerns a 72-year-old male from Mexico who had a history of type 2 diabetes mellitus. Sequencing of the partial ribosomal operon revealed that in both cases a member of the order Pleosporales was concerned which could not be affiliated to any family within this order. Multilocus analysis revealed that the fungus was related to another, unaffiliated agent of human mycetoma, Pseudochaetosphaeronema larense, and therefore the name Pseudochaetosphaeronema martinelli was introduced.

Identification and ultrastructural characterization of the Wolbachia symbiont in Litomosoides chagasfilhoi.

BACKGROUND: Filarial nematodes are arthropod-transmitted parasites of vertebrates that affect more than 150 million people around the world and remain a major public health problem throughout tropical and subtropical regions. Despite the importance of these nematodes, the current treatment strategies are not efficient in eliminating the parasite. The main strategy of control is based on chemotherapy with diethylcarbamazine, albendazole and ivermectin. In the 1970s, it was found that some filarids possess endosymbiotic bacteria that are important for the development, survival and infectivity of the nematodes. These bacteria belong to the genus Wolbachia, which is a widespread and abundant intracellular symbiont in worms. Knowledge about the structure of the bacteria and their relationship with their nematode hosts may allow new perspectives for the control of filarial nematodes. METHODS: In this study, we used transmission electron microscopy combined with three-dimensional approaches to observe the structure of the endosymbiont of the filarial nematode Litomosoides chagasfilhoi, an experimental model for the study of lymphatic filariasis. In addition, the bacterium was classified based on PCR analyses. RESULTS: The bacterium was mainly found in the hypodermis and in the female reproductive system in close association with host cell structures, such as the nucleus and endoplasmic reticulum. Our ultrastructural data also showed that the symbiont envelope is composed of two membrane units and is enclosed in a cytoplasmic vacuole, the symbiosome. Molecular data revealed that the bacterium of L. chagasfilhoi shares 100% identity with the Wolbachia endosymbiont of Litomosoides galizai. CONCLUSIONS: Here we described ultrastructural aspects of the relationship of the Wolbachia with the filarial nematode Litomosoides chagasfilhoi and the findings lead us to consider this relationship as a mutualistic symbiosis.

Molecular identification of melanised non-sporulating moulds: a useful tool for studying the epidemiology of phaeohyphomycosis.

Subcutaneous infections caused by melanised fungi have been increasingly reported among transplant patients, and these infections have the potential for blood and visceral dissemination. Some moulds, such as Mycelia sterilia, cannot grow and sporulate on different media, making their identification impossible by conventional methods. The fast and accurate identification of melanised fungi at the species level is important because species may have tropism to different organs and different susceptibilities to antifungal agents. Molecular tools have been reported to be helpful for the species identification of non-sporulating moulds. Our goal was to identify the species of M. sterilia isolates obtained from clinical samples of transplant patients using sequences of ITS and the D1/D2 regions of rDNA. Clinical samples were obtained from eight kidney transplant recipients who developed subcutaneous fungal infections. The diagnosis was confirmed by histopathology and conventional culture. Histopathology showed septated, melanised hyphae, and the cultures identified non-sporulating fungi. Therefore, the DNA from the M. sterilia isolates was subjected to PCR amplification and sequencing of the ITS and D1/D2 regions. Genus/species identification was obtained by comparison with gene banks. We obtained the following identifications: Alternaria sp. (2), Cochliobolus lunatus/Curvularia lunata (2), Cochliobolus hawaiiensis/Bipolaris hawaiiensis (1), Ochroconis sp. (1), Medicocopsis romeroi/Pyrenochaeta romeroi (1) and Nigrograna mackinnonii/Pyrenochaeta mackinnonii (1).

Classification of subcutaneous and systemic mycoses.

The classification of human fungal infections in medical reports is sometimes confusing. This occurs because some agents act as opportunistic organisms in immunosuppressed patients, whereas others affect the subcutaneous tissue and also cause disseminated or systemic disease. Finally, some clinically similar infections caused by aerobic actinomycetic bacteria and others caused by parasites (rhinosporidiosis) have been traditionally included in the descriptions of mycotic diseases. This contribution provides the clinician with a classification of subcutaneous, systemic, and opportunistic fungal infections.

Subcutaneous phaeohyphomycosis (mycotic cyst).

Phaeohyphomycosis designates fungal infections caused by pheoid or melanized fungi and characterized histopathologically by the presence of septate hyphae, pseudohyphae, and yeasts. Etiologic agents include Exophiala, Phoma, Bipolaris, Phialophora, Colletotrichum, Curvularia, Alternaria, Exserohilum, and Phialemonium sp. The most common are Exophiala jeanselmei and Wangiella dermatitidis. The clinical presentation depends on the immune status of the host: superficial (tinea nigra and black piedra); cutaneous (scytalidiosis) and corneal; subcutaneous (mycotic cyst); and systemic phaeohyphomycosis in the immunocompromised host. The mycotic cyst is a localized form, characterized by subcutaneous asymptomatic nodular lesions that develop after traumatic implantation of fungi, especially on the extremities. The average size of the cysts is 2.5 cm. KOH examination reveals pigmented yeasts, pseudohyphae, and hyphae. A cutaneous biopsy specimen usually shows an abscess or a suppurative granuloma with pigmented yeasts and pseudohyphae. The treatment of choice is surgical excision, but additional anti-fungal therapy is recommended for recurrent cases and immunocompromised patients.

Host defenses in subcutaneous mycoses.

Subcutaneous mycoses include diverse clinical syndromes, characterized by invasion of the skin and subcutaneous tissue by saprobic fungi. Individuals living in rural areas constantly suffer lesions or trauma; however, only a few of them develop disease. In this contribution, we describe recent advances in the understanding of the virulence of these organisms, focusing on the most prevalent infections, sporotrichosis, chromoblastomycosis, and mycetoma. Although these infectious diseases are considered neglected tropical diseases, modern molecular techniques have been able to identify the etiologic agents and observe variations in the former monolithic concept of the species, which was based mostly on morphologic characteristics. The complete genetic characterization of the causative agents, along with that of their host, will help in the understanding of the factors on which the development of these infections depends.

Mycoses of implantation in Latin America: an overview of epidemiology, clinical manifestations, diagnosis and treatment.

Implantation or subcutaneous mycoses are a frequent health problem in Latin American countries and other tropical and subtropical areas. Although such infections rarely cause disseminated or invasive disease, they have an important impact on public health, and timely diagnosis and appropriate treatment remain important. Although some implantation mycoses are found in immunocompromised persons, the immunocompetent population is the principal target in Latin America. Most etiologic agents are found in soil, vegetation, and decaying matter in tropical, subtropical, and humid environments and infection is commonly the result of penetrating injury. Infections primarily occur (1) among low socioeconomic groups, (2) among those living in rural areas or involved in farming, hunting, or other outdoor activities, and (3) particularly among adult men. This review focuses on the epidemiology of the most clinically significant implantation mycoses in Latin America, i.e., sporotrichosis, eumycetoma, chromoblastomycosis, subcutaneous phaeohyphomycosis, subcutaneous zygomycosis, and lacaziosis. Main epidemiologic findings, clinical manifestations, diagnosis, and treatment options are also discussed.

Subcutaneous phaeohyphomycosis.

This report shows images of a case of subcutaneous phaeohyphomycosis caused by Exophiala jeanselmei in a patient who has undergone renal transplantation. The paper comments briefly on the disease and emphasizes the need to take this mycosis into account in the differential diagnosis of other dermatoses, including non-infectious dermatoses.

Feoifomicose subcutânea / Subcutaneous phaeohyphomycosis

São apresentadas imagens ilustrativas de um caso de feoifomicose subcutânea causada pela Exophiala jeanselmei num paciente transplantado renal. Breves comentários sobre a doença encontram-se no texto. Ressalta-se a necessidade de essa micose entrar no diagnóstico diferencial de outras dermatoses, inclusive as não infecciosas.

This report shows images of a case of subcutaneous phaeohyphomycosis caused by Exophiala jeanselmei in a patient who has undergone renal transplantation. The paper comments briefly on the disease and emphasizes the need to take this mycosis into account in the differential diagnosis of other dermatoses, including non-infectious dermatoses.

Cutaneous and subcutaneous phaeohyphomycosis.

UNLABELLED: Case 1: A 17-year-old male rural worker from Bolivia living in La Plata (Argentina) for the past year had a lesion on the flexor side of his right forearm (6 x 4 cm). The lesion was formed by several confluent nodular areas, wine-red in color, some fistulized, with hemopurulent drainage. The area was hot and painless (Figure 1). On physical examination, no regional adenomegalies were reported. The following analyses were requested and results reported. Soft tissue ultrasound: material of solid consistency with layered liquid areas, located in the subcutaneous cellular tissue, with fistulous tract, connecting through superficial planes. Evidence of peripheral edema. Bacteriologic analysis (puncture aspiration): methicillin-sensitive Staphylococcus aureus. Mycologic analysis (puncture aspiration): negative; laboratory results: eosinophilia; and human immunodeficiency virus: nonreactive. Histopathologic examination: lesions of necrosis with granulomatous inflammatory reaction. Fungi techniques (periodic acid-Schiff, Grocott stains): negative. Bacilos acid-alcohol resistentes (acid-alcohol resistant bacillus) (BAAR) techniques (Kinyoun, Ziehl-Neelsen): negative. Foreign body examination tested with polarized light: negative. Mycologic and bacteriologic examinations were repeated, including a search for mycobacterium species using material obtained from the biopsy performed on the cutaneous lesion. Macromorphology: the colony was initially black and of creamy consistency, to later become velvety. Micromorphology: dark blastoconidia, then cylindrical phialides with elliptical conidia (Figure 2). IDENTIFICATION: Exophiala dermatitidis infection. On the basis of these characteristics, the diagnosis is phaeohyphomycosis due to Edermatitidis. The patient is treated with antimycotic therapy, with oral itraconazole (400 mg/d), plus indication of surgical procedure to remove the lesion. The patient's condition evolves favorably with no recidivant episodes after the sixth month post-treatment (Figure 3). During the first year, controls were scheduled every 2 months. Case 2: A 72-year-old diabetic man had a painful chronic varicose ulcer on the side of his left foot, with black friable exudate, 2x3 cm in diameter after 1 year. Every time the black material was removed, it would quickly grow back again. No response was obtained with different therapies applied to seal the lesion (Figure 4). Routine laboratory results included the following. Glucemy: 1.82 g/dL. Histopathology: filamentous septate fungal elements with positive Grocott stain (Figure 5 and Figure 6). Mycologic examination and culture: direct: fungal elements in dematiaceous group. Culture: positive for Curvularia lunata (Figure 7). The treatment selected was oral itraconazole (400 mg/d) for 12 months, with periodic laboratory controls, plus application of wet pads on the ulcer containing sodium borate and ketoconazole cream. At the fourth month, the ulcer had completely closed (Figure 8).

Is TGF-beta important for the evolution of subcutaneuos chronic mycoses?

Jorge Lobo's and chromoblastomycosis are chronic deep mycosis that clinically manifests as keloid and verrucous polymorphic lesions of solid consistency and variable size that contain small scales and crusts. Few studies are available in the literature characterizing the in situ cellular and humoral immune response, especially the involvement of cytokines which immunosuppressive and fibrogenic effects as the TGF-beta. The hypothesis of the present paper is explaining the possible mechanism of this cytokine in cutaneous lesions pathology in chromoblastomycosis and lacaziosis (Jorge Lobo's disease). The results of this investigation are a new hypothesis for ethiopatogenesis of these diseases: TGF-beta is a double effect that follows fibrosis and immunosuppression in local skin.

Subcutaneoous phaeohyphomycosis by Exophiala jeanselmei in a cardiac transplant recipient.

We report a case of phaeohyphomycosis caused by Exophiala jeanselmei in a cardiac transplant recipient maintained on immunosuppressive therapy with mycophenolate mofetil tacrolimus and prednisone. The lesion began after trauma on the right leg that evolved to multiple lesions with nodules and ulcers. Diagnosis was performed by histological examination and culture of pus from skin lesions. Treatment consisted of itraconazole (200 mg/day) for three months with no improvement and subsequently with amphotericin B (0.5 mg/Kg per day to a total of 3.8 g intravenously). After four months of treatment, the lesions showed marked improvement with reduction in the swelling and healing of sinuses and residual scarring.

Subcutaneoous phaeohyphomycosis by Exophiala jeanselmei in a cardiac transplant recipient

Este trabalho relata um caso de feohifomicose subcutânea causado por Exophiala jeanselmei em um paciente que havia recebido transplante de coração e mantinha terapia com micofenolato mofetil, tracolimus e prednisone. As lesões tiveram início após trauma na perna inferior direita que evoluíram produzindo múltiplos nódulos e úlceras. Diagnóstico foi realizado através de avaliação histológica e de características macroscópicas e microscópicas da cultura das lesões da pele. O paciente fez uso de itraconazol em concentração de 200 mg/dia durante três meses, não se observando no entanto, melhora das lesões. Após este período, o paciente foi tratado com anfotericina B a uma concentração de 0,5 mg/Kg/dia totalizando 3,8 g. Após quatro meses de tratamento as lesões mostraram melhora evidente, verificando-se fechamento das fístulas e cicatrização das lesões.

Subcutaneous phaeohyphomycosis caused by Lasiodiplodia theobromae and successfully treated surgically.

While visiting Jamaica, a 50-year-old woman stumbled on an outdoor wooden staircase and sustained an injury to the right leg. The wound was cleaned topically and the patient was given antibacterial therapy. Five weeks later, in Canada, she presented with an ulcer at the injury site. An excisional biopsy showed copious broad, septate, melanized fungal filaments penetrating into tissue. Culture yielded a nonsporulating melanized mycelium. The isolate was strongly inhibited by cycloheximide and benomyl but grew at 37 degrees C. After 16 weeks cultivation on modified Leonian's agar at 25 degrees C, it developed pycnidia characteristic of Lasiodiplodia theobromae, a common tropical phytopathogen mainly known previously as a rare agent of keratitis and onychomycosis in humans. The patient was not given antifungal chemotherapy, and the ulcer, which had been broadly excised in the biopsy procedure, ultimately resolved after treatment with saline compresses. The six-month follow-up showed no sign of infection. This case, interpreted in light of previously reported cases, shows that on rare occasions L. theobromae is able to act as an agent of subcutaneous phaeohyphomycosis and that, when this occurs, debridement alone may be sufficient to eradicate it.