Examples of bioactive peptides derived from internal sequences of proteins are known for decades. The great majority of these findings appear to be fortuitous rather than the result of a deliberate and methodological-based enterprise. In the present work, we describe the identification and the biological activities of novel antimicrobial peptides unveiled as internal fragments of various plant proteins founded on our hypothesis-driven search strategy. All putative encrypted antimicrobial peptides were selected based upon their physicochemical properties that were iteratively selected by an in-house computer program named Kamal. The selected peptides were chemically synthesized and evaluated for their interaction with model membranes. Sixteen of these peptides showed antimicrobial activity against human and/or plant pathogens, some with a wide spectrum of activity presenting similar or superior inhibition efficacy when compared to classical antimicrobial peptides (AMPs). These original and previously unforeseen molecules constitute a broader and undisputable set of evidences produced by our group that illustrate how the intragenic concept is a workable reality and should be carefully explored not only for microbicidal agents but also for many other biological functions.

Introduction: In this study, the clinical features, underlying diseases and clinical outcomes of patients with cryptococcosis were investigated. In addition, a molecular analysis of the Cryptococcus neoformans species complex isolated from these patients was performed. Methods: A prospective study of 62 cases of patients with cryptococcal infection was conducted at the Hospital de Doenças Tropicais de Goiás Dr. Anuar Auad from 2009-2010. Cryptococcal meningitis cases were diagnosed by direct examination and cerebrospinal fl uid (CSF) sample culture. The profi ling of these patients was assessed. The CSF samples were submitted to India ink preparation and cultured on Sabouraud dextrose agar, and C. neoformans was identifi ed by the production of urease, a positive phenoloxidase test and assimilation of carbohydrates. C. neoformans and C. gattii isolates were distinguished by growth on L-canavanine-glycine-bromothymol blue medium, and molecular analysis was conducted via PCR fi ngerprinting reactions using M13 and (GACA) 4 primers. Results: From the 62 patients with cryptococcosis, 71 isolates of CSF were obtained; 67 (94.4%) isolates were identifi ed as C. neoformans var. grubii/VNI, and 4 (5.6%) were identifi ed as C. gattii/VGII. Of these patients, 53 had an HIV diagnosis. The incidence of cryptococcosis was higher among patients 20-40 years of age, with 74.2% of the cases reported in males. Cryptococcus-related mortality was noted in 48.4% of the patients, and the symptoms were altered sensorium, headache, fever and stiff neck. Conclusions: The high morbidity and mortality observed among patients with cryptococcosis demonstrate the importance of obtaining information regarding the epidemiological profi le and clinical course of the disease in the State of Goiás, Brazil.

Following the treads of our previous works on the unveiling of bioactive peptides encrypted in plant proteins from diverse species, the present manuscript reports the occurrence of four proof-of-concept intragenic antimicrobial peptides in human proteins, named Hs IAPs. These IAPs were prospected using the software Kamal, synthesized by solid phase chemistry, and had their interactions with model phospholipid vesicles investigated by differential scanning calorimetry and circular dichroism. Their antimicrobial activity against bacteria, yeasts and filamentous fungi was determined, along with their cytotoxicity towards erythrocytes. Our data demonstrates that Hs IAPs are capable to bind model membranes while attaining α-helical structure, and to inhibit the growth of microorganisms at concentrations as low as 1μM. Hs02, a novel sixteen residue long internal peptide ( ) derived from the unconventional myosin 1h protein, was further investigated in its capacity to inhibit lipopolysaccharide-induced release of TNF-α in murine macrophages. Hs02 presented potent anti-inflammatory activity, inhibiting the release of TNF-α in LPS-primed cells at the lowest assayed concentration, 0.1 μM. A three-dimensional solution structure of Hs02 bound to DPC micelles was determined by Nuclear Magnetic Resonance. Our work exemplifies how the human genome can be mined for molecules with biotechnological potential in human health and demonstrates that IAPs are actual alternatives to antimicrobial peptides as pharmaceutical agents or in their many other putative applications.

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Introduction: This is the fi rst study to examine the in vitro susceptibility and the expression of virulence factors in Candida species in the presence of Pimenta pseudocaryophyllus (Gomes) L.R. Landrum (Myrtaceae), a Brazilian plant known as paucravo. Additionally, the mechanisms of action of the crude ethanol extract and the ethyl acetate and aqueous fractions of this plant were investigated. Methods: The in vitro susceptibility of Candida was tested using the broth microdilution method, whereas an XTT reduction assay was used for biofi lms. Adherence was determined by counting the number of yeast cells that adhered to 100 oral epithelial cells, and hyphal formation was verifi ed in the hyphal induction medium M199. Flow cytometry with propidium iodide and FUN-1 was performed to assess the mechanism of action. Results: The results revealed that the crude ethanol extract and the ethyl acetate and aqueous fractions of P. pseudocaryophyllus inhibited the growth of Candida isolates at a minimal inhibitory concentration (MIC) ranging from 64 to 256µg/mL, whereas the 50% sessile minimal inhibitory concentration (SMIC 50 ) ranged from 512 to >1,024µg/mL. Adherence and hyphal formation were signifi cantly reduced in the presence of the crude ethanol extract and both fractions. Although cell membrane injury was detected, the predominant mechanism of action appeared to be the alteration of yeast metabolism, as demonstrated by fl ow cytometry. Conclusions: Our results indicated that antifungal activity reduced the expression of virulence factors in yeast via the alteration of yeast metabolism, suggesting that the crude extract of P. pseudocaryophyllus and its fractions may contain novel antifungal agents.

The aim of this study was to perform from the morpho-anatomical study, physicochemical characterization, chemical composition, seasonal variability and larvicidal activity of the volatile oils of Campomanesia pubescens (DC.) O. Berg leaves. The botanical material was collected in Hidrolândia, Goiás. Morpho-anatomical characterization and phytochemical screening were performed by conventional methods. Leaf powders were submitted to hydrodistillation in the Clevenger apparatus and the identification of the chemical components of the volatile oils obtained was done by GC-MS. The larvicidal activity was carried out with third-stage larvae of the Aedes aegypti. The leaf blade, the main vein, and the young stem have a uni-stratified epidermis covered by a cuticle. The mesophyll and cortical parenchyma of the main vein and young stem have secretory cavities and idioblasts with crystals. The main rib contains a bicolateral bundle in an open arch. The young stem has a discontinuous band of sclerenchyma external to the phloem cells. Most volatile oil compounds are spathulenol, caryophyllene oxide, α-macrocarpene, and z-caryophylleneIn phytochemical analysis, tannins, digitalis, flavonoids, and total phenols were detected. The content of volatile compounds was 7.36%, that of total ash was 1.77%, and that of mucilage was 3.52 ml. The volatile oil at the concentration used was inactive against Ae aegypti larvae. The present study contributes to the taxonomic knowledge of the species and provides parameters for quality control of the plant raw material. This work represents the first description of the chemical compounds and seasonal variability of volatile oils from C. pubescens leaves collected in Goiás state.

doi: 10.5216/ree.v12i3.6472As espécies de Candida são de ocorrência comum na cavidade bucal sendo que estas leveduras estão presentes em cerca de 50% de indivíduos saudáveis, podendo este porcentual variar dependendo da população estudada. Os processos infecciosos causados por Candida variam de quadros clínicos benignos, a quadros graves e fatais como as infecções invasivas e disseminadas. Neste estudo foi avaliada a colonização de leveduras do gênero Candida em 312 amostras de saliva de profissionais do Hospital das Clínicas da Universidade Federal de Goiás, na cidade de Goiânia, Goiás, Brasil, durante o período de julho a dezembro de 2007. As espécies de Candida isoladas da saliva foram identificadas através da produção de tubo germinativo e de clamidoconídios, assimilação de carboidratos e cultivo em meio CHROMagar® Candida. Os profissionais estudados mostraram colonização por leveduras do gênero Candida em 61,9% das amostras analisadas, sendo isolados C. albicans (58,5%), C. parapsilosis (24,4%), C. krusei (8,3%) e C. famata (3,6%). A divulgação destes resultados pode servir como um auxílio para a prevenção e controle de candidíase bucal nos trabalhadores da área de saúde que vivem em contato constante com indivíduos altamente suscetíveis a infecções.Descritores: Cândida; Saliva; Pessoal de saúde.

O envelhecimento populacional tem crescido nas últimas décadas e, com isso, aumentou também a incidência das doenças crônicas, como é o caso da Hipertensão Arterial Sistêmica (HAS). A HAS é uma doença assintomáticacaracterizada pela elevação persistente dos níveis da Pressão Arterial, iguais ou acima de 140/90 mmHg e sua detecção é feita após repetidas aferições. A doença é um problema de saúde pública no Brasil, pois tem grande prevalência e está associada a diversas patologias como acidente vascular cerebral (AVC), infarto do miocárdio e insuficiência renal. O controle da HAS é feito por meio de mudanças no estilo de vida e uso de medicamentos, no entanto, a administração dos fármacos, pode trazer diversos riscos, especialmente para os idosos. Diante disso, tornou-se necessário investigar a importância da Atenção Farmacêutica no acompanhamento dos pacientes idosos e hipertensos, no Brasil, que é a proposta do presente estudo.