Effects of artepillin C on model membranes displaying liquid immiscibility

It has been hypothesized that the therapeutic effects of artepillin C, a natural compound derived from Brazilian green propolis, are likely related to its partition in the lipid bilayer component of biological membranes. To test this hypothesis, we investigated the effects of the major compound of green propolis, artepillin C, on model membranes (small and giant unilamelar vesicles) composed of ternary lipid mixtures containing cholesterol, which display liquid-ordered (lo) and liquid-disordered (ld) phase coexistence. Specifically, we explored potential changes in relevant membrane parameters upon addition of artepillin C presenting both neutral and deprotonated states by means of small angle X-ray scattering (SAXS), differential scanning calorimetry (DSC), and confocal and multiphoton excitation fluorescence microscopy. Thermotropic analysis obtained from DSC experiments indicated a loss in the lipid cooperativity of lo phase at equilibrium conditions, while at similar conditions spontaneous formation of unilamellar vesicles from SAXS experiments showed that deprotonated artepillin C preferentially located at the surface of the membrane. Time-resolved experiments using fluorescence microscopy showed that at doses above 100 µM, artepillin C in its neutral state interacted with both liquid-ordered and liquid-disordered phases, inducing curvature stress and promoting dehydration at the membrane interface.

Cladosporium cladosporioides isolado de gato com carcinoma de células escamosas / Cladosporium cladosporioides isolated from a cat with squamous cell carcinoma

Este relato descreve o caso do Cladosporium cladosporioides isolado de uma lesão periocular de um felino atendido no Hospital Veterinário da Universidade Federal de Mato Grosso, em Cuiabá. A principal queixa do proprietário era uma lesão periocular, com piora no decorrer do tempo, havia aproximadamente quatro meses. Foi descrita a tentativa de tratamento da lesão com anti-inflamatórios e antibióticos, sem sucesso. Na anamnese foi relatado que o animal tinha acesso à rua e a hábitos de caça e que não havia outros animais da casa com lesão semelhante. O animal foi submetido à biópsia e citologia para um diagnóstico mais preciso do caso. Um fragmento foi encaminhado para o Laboratório de Patologia Veterinária e outro para o de Microbiologia Veterinária. Nas análises histopatológicas, houve compatibilidade com carcinoma de células escamosas e, nas lâminas de citologia, foi evidenciado um processo inflamatório agudo. Nas características macroscópicas e microscópicas da colônia, houve compatibilidade com Cladosporium sp. Posteriormente, o DNA foi extraído e sequenciado, confirmando a espécie Cladosporium cladosporioides. O objetivo deste relato foi descrever o isolamento dessa espécie em um felino com carcinoma de células escamosas.(AU)

This report describes a case of Cladosporium cladosporioides isolated from a cat with a periocular lesion at the Veterinary Hospital, Cuiaba- Brazil. Owner described his animal as having a periocular lesion treated unsuccessfully with anti-inflamatories and antibiotics. During anamnesis, it was reported that the animal has access to the street, hunting habits and none of the other animals of the house had a similar injury. The animal underwent biopsy and cytology for more accurate diagnosis of the case. A fragment was referred to the Veterinary Pathology Laboratory and another for Veterinary Microbiology. In the histopathological analysis of biopsy, it was compatible with squamous cell carcinoma and the cytology slides showed an acute inflammatory process. Microbiogical analysis isolated fungus with Cladosporium sp. Subsequently, DNA was extracted and sequenced confirming Cladosporium cladosporioides species. This paper reports the isolation of this species in a feline with squamous cell carcinoma.(AU)

Detection of echinococcosisinfections in Mongolia using ELISA with recombinant AgB and Em18 antigens / Монголын Анагаах Ухаан

Background. Echinococcosis are parasitic zoonoses, remains a public health problem of worldwide, including Mongolia . Differential diagnosis between E.granulosus and E.multilocularis has significant implications for epidemiologic studies, treatment of these diseases, since both species occur in Mongolia. Serodiagnostic tests based on detection of antibodies against genus and species-specific antigens have played an important role in differential diagnosis, confirming clinical diagnosis and in epidemiologic studies.Materials and Methods. A total of 107 volunteer participants’ serum samples and additional 11 serum samples from the persons with hepatic cysts were tested for specific IgG against recombinant AgB and recombinant Em18 antigens in an ELISA .Results.rAgB-specific antibody was detected in 2 (3.33) of 60 individuals from Bayankhongor province and no one had positive response to this antigen in 46 individuals from Ulaanbaatar city while rEm18-specific antibody was present in 7 (11.66) and 3 (6.38) respectively. The one individual with typical lesions of active echinococcosis in a liver revealed by abdominal ultrasonography showed significantly higher IgG antibody response to rAgB. We suggest that people need to be provided information not only about cystic echinococcosis but also alveolar echinococcosisand improvement of sanitation and hygiene and to be careful with corsac and red foxes and their feces to prevent those infections.

Cysticercosis/taeniasis: recent advances in serological and molecular diagnoses.

Serodiagnosis by immunoblot, using recombinant chimeric T. solium antigen and native glycoprotein antigens, has been applied for neurocysticercosis cases. Specific antibodies against both antigens were detected in serum samples from NCC patients involving multiple cysts in the brain, whereas it was not always easy to detect specific antibodies in NCC cases with a solitary cyst or calcified lesion(s). On the other hand, the diagnosis for human taeniasis or worm carriers has been routinely performed by stool examination. In this study, multiplex PCR has been established to differentiate taeniasis using Taenia mitochondrial DNA in fecal samples from worm carriers. Furthermore, the molecular identification of human taeniid cestodes by base excision sequence scanning thymine-base analysis has also been introduced. This method provides four thymine-base peak profiles unique for Asian and American/African genotypes of T. solium, T. saginata and T. asiatica. By comparing thymine base peak profiles, it is possible to differentiate human taeniid cestodes without DNA sequencing. The approaches are powerful tools for the routine diagnosis of taeniasis and the molecular identification of taeniid cestodes.

Recent advances in basic and applied science for the control of taeniasis/cysticercosis in Asia.

Detection of seven specific bands by immunoblot (IB) using glycoproteins (GPs) purified by lentil-lectin affinity chromatography has been the gold-standard for neurocysticercosis (NCC) serodiagnosis since 1989. However, due to the presence of contaminants, it was impossible to apply the GPs to ELISA. Our group at Asahikawa Medical College (AMC) succeeded in purifying the GPs by preparative isoelectric focusing; these higher quality GPs were suitable for ELISA. Based on the results of both IB and ELISA testing, developed at AMC for a field survey in Irian Jaya, it became evident that that area had pandemic NCC. We found many NCC patients, pigs full of cysts, and one dog infected with two cysts: these findings were based on serology. Recently, we conducted another survey to detect of the worm carriers of T. solium. Three of the 38 local people were positive by copro-antigen specific to Taenia species; these three patients expelled segments of Taenia spp and these were confirmed as those of T. solium by mitochondrial DNA analysis. When viable eggs of any taeniid species could be obtained, they can be developed into metacestodes in NOD-scid mice; it then becomes possible to analyze morphological dynamics, metacestode antigenicity, the efficacy of new metacestocidal drugs, and mitochondrial DNA. Mitochondrial DNA analysis of the specimens obtained in Irian Jaya was compared with that of other isolates worldwide. T. solium is now divided into two genotypes: the Asian type, and the Africa-American type. Some aspects of the pathological differences between the Asian and Africa-American types and the antigenic components of these two types are discussed.

NOD-scid mouse as an experimental animal model for cysticercosis.

The major three species of human taeniid cestodes, Taenia solium, T. saginata and T. saginata asiatica (= T. asiatica) which require humans as the definitive host are still not rare in developing countries. Among these, T. solium is the most serious with medical and economic importance. Neurocysticercosis (NCC) in humans is now recognized as the major cause of neurologic disease in the world. As these human taeniid cestodes obligatory require domestic animals such as swine, cattle and swine as the major intermediate host animals respectively, it is not easy to analyze the basic research in these domestic animals. In this brief review, we introduce experimental animal model for these three species in order to obtain fully developed metacestode stage in severe combined immunodeficiency (scid) mice. Non-obese diabetic scid (NOD-scid) mice are expected to be a satisfactory animal model and to have advantages for analysis by several view points of developmental biology with gene expression throughout development, antigenic homology of cyst fluid of these three species, evaluation of drug efficacy or metacestocidal drug designs, confirmation of unknown taeniid gravid segments for identification based on the morphology and DNA analysis of metacestodes. The animal model is not only available for human Taenia spp but can also be applied to other taeniid cestodes of economic importance or in veterinary parasitology.

Recent advances in serodiagnosis for cysticercosis.

Neurocysticercosis (NCC) caused by infection with the larval stage of Taenia solium is an important cause of neurological disease worldwide. Up to the present, many studies on characterizing species-specific antigens of T. solium have been done and several high quality antigens for serodiagnosis are available. Hence the research on serodiagnosis has been shifted to the next phase, stable production of diagnostic antigens using molecular techniques. In order to establish an enzyme-linked immunosorbent assay (ELISA) using recombinant proteins, we carried out molecular cloning and identified four diagnostic antigen candidates (Ag1, Ag1V1, Ag2, and Ag2V1). Recombinant proteins, except Ag2V1, were successfully expressed using an Escherichia coli expression system. Immunoblot analysis using NCC patient sera detected recombinant proteins. But as reactivity to rAg1 was too weak, Ag1 was not suitable for the immunodiagnosis antigen. Therefore Ag1V1 and Ag2 were chosen for ELISA antigens and Ag1V1/Ag2 chimeric protein was expressed. Of 49 serum samples from NCC patients confirmed to be seropositive by immunoblot analysis, 44 (89.7%) were positive by ELISA. Serum samples from patients with other parasitic infections did not recognized Ag1V1/Ag2 chimeric protein. Ag1V1/Ag2 chimeric protein obtained in this study is of value for differential immunodiagnosis.

Molecular variation of Taenia solium in the world.

Complete nucleotide sequences of the mitochondrial cytochrome b (Cytb) and cytochrome c oxidase subunit I (CO I) genes from various isolates of Taenia solium were examined. Eleven isolates were analyzed; two isolates from China, two isolates from Indonesia, one isolate each from India, Thailand, Mexico, Ecuador, Peru, Mozambique and Tanzania. In both genes, two isolates from Indonesia shared the same sequences. Similarly, the isolate from Mexico shared same sequences with that from Peru, and the isolate from Mozambique shared same sequences with that from Tanzania. Phylogenetic trees inferred from different mitochondrial genes yielded almost the same topology. Both the UPGMA and NJ-trees were also very similar. These trees indicate that T. solium may be diverged to 2 genetic groups; isolates from Asia form one group and isolates from Africa and Latin America belong to the other. It seems that T. solium prevalent in Africa and in Latin America shares the related origin and has recently been introduced to each area, perhaps with domestic pigs or human.

Differential serodiagnosis of cystic and alveolar echinococcosis using native and recombinant antigens in Japan.

Our group at Asahikawa Medical College has established differential serodiagnosis for zoonotic larval cestodiases such as alveolar echinococcosis (AE), cystic echinococcosis (CE) and neurocysticercosis (NCC) using purified specific antigens. In this brief review, we introduce (a) four imported CE cases in Japan, easily identified serologically, (b) most recent advances in serology for differentiation of AE and monitoring of prognosis of AE in Japan. It includes application of affinity purified Em18 and prototype of a recombinant Em18 antigen. Serology using affinity purified Em18 antigens is showing much higher sensitivity for detection of AE cases which are usually undetectable by the ongoing serology for AE authorized in Hokkaido, Japan. As serology for AE, CE or NCC is still not popular in the majority of Asian countries, we expect that this review paper stimulates researchers who are interested in serology or serodiagnosis for these larval cestodiases including AE, CE and NCC.

Cysticercosis in Indonesia: epidemiological aspects.

Cysticercosis caused by the larval stage, cysticercus or cysticerci, of the pork tapeworm Taenia solium was recognized at first in Bali and in Paniai District, Irian Jaya (Papua), Indonesia in the 1970s. In the 1990s a rapid increase in the number of the cases of epileptic seizures and burns in Jayawijaya district, eastern Papua, was observed. There were a total of 1,120 new cases of burns (7.0%) and 293 new cases of epileptic seizures (1.8%) from 15,939 local people during 1991-1995. Both histopathological examination and mitochondrial DNA analysis of resected cysts from patients and pigs revealed cysticerci of T. solium. Antibody responses highly specific to cysticercosis were revealed in approximately 67% and 65% of persons respectively with epileptic seizures and with subcutaneous nodules. Therefore, most cases of epileptic seizures and burns were considered to be associated with cysticercosis in Papua. Additional serologically data from Bali showed that 13.5% of epileptic seizures (10/74) and 12.6% of asymptomatic individuals (94/746) were supposed having been exposed to T. solium. Histopathological evaluation of 80,000 tissue samples in East Java revealed that nine were cysticercosis. All cases were non-moslems and from two ethnic groups, Chinese and Balinese. Epidemiological data on cysticercosis are not available from other provinces of Indonesia, although cases of cysticercosis are occasionally reported. Therefore, other intensive epidemiological studies are strongly recommended, especially covering the eastern part of Indonesia.

Neurocysticercosis and echinococcosis in Asia: recent advances in the establishment of highly reliable differential serodiagnosis for international collaboration.

Neurocysticercosis (NCC) and echinococcosis, caused by the larval stage of taeniid cestodes, are recognized as major parasitic zoonoses threatening human life worldwide. Cystic echinococcosis (CE), caused by Echinococcus granulosus, has well been known to be more widely distributing in Europe and Asia (Eurasia) than alveolar echinococcosis (AE) caused by E. multilocularis. However, it has recently been found that AE is more widely distributing or spreading in Eurasia. Furthermore, NCC caused by Taenia solium is also spreading in Eurasia. Due to the lack of reliable methodology for diagnosing these zoonotic cestodiases worldwide, prevalence rates of these diseases are extremely underestimated. Our group has been working for the establishment of differential serodiagnosis and molecular diagnosis of AE. CE and NCC as international collaboration projects sponsored by the Ministry of Education, Japan from 1994 until 2000 at least. In this paper, we introduce (1) the most recent original work on the establishment of differential serodiagnoses of NCC, AE and CE, (2) international collaboration work on epidemiology of these diseases in several countries, and discuss (3) what we can and should do for the control of such global parasitic diseases. It is stressed that international collaboration or cooperation work on the control of parasitic diseases is only successful based on the original scientific contribution of high standard.

Desenvolvimento de um novo modelo acadêmico na educaçäo dos profissionais de saúde no contexto do Prouni-Londrina (1991-1997): sistematizaçäo e reflexöes teórico-metodológicas / Development of a new academic model in the pedagogical support of health professionals in the Prouni-Londrina context (1991-1997): sistematization and reflexions

Este trabalho consiste na análise e reflexäo das transformaçöes que vêm sendo realizadas nos últimos 6 anos nos cursos do Centro de Ciências da Saúde da Universidade Estadual de Londrina. Em 1991, foram regulamentadas mudanças de regime acadêmico de crédito para seriado, e de currículos, além da seleçäo do CCS para participar do programa UNI. Desde entäo, várias estratégias vem sendo utilizadas com vistas ao novo modelo acadêmico, tais como a implantaçäo de projetos especiais que integram academia, serviços de saúde e comunidade, projetos de apoio ao desenvolvimento de pesquisas integradas ao ensino e à comunidade, criaçäo de estrutura de apoio pedagógico e mudanças na estrutura organizacional, além do processo de aprimoramento e capacitaçäo em novas metodologias, sempre com uma grande participaçäo de docentes, alunos e membros da comunidade. Em vista deste processo de construçäo coletiva, tem início a definiçäo dos projetos pedagógicos dos 5 cursos do CCS, baseados nos problemas relevantes da sociedade, nas tendências internacionais de educaçäo e com estratégias/metodologias de ensino que privilegiam a construçäo do conhecimento. Para a reformulaçäo, aprimoramento e redirecionamento das nossas metas, nos baseamos em pressupostos teóricos da transdisciplinaridade, e na teoria da complexidade, rumo a Pedagogia da Interaçäo