Assessment of electrochemotherapy effects on the development of Ehrlich solid tumor in swiss mice using a novel electroporator device / Avaliação dos efeitos da eletroquimioterapia sobre o desenvolvimento do tumor sólido de Ehrlich em camundongos Swiss utilizando um novo eletroporador

Electrochemotherapy is a local anticancer treatment in which non-permeant chemotherapeutic drugs are associated with electric pulses of well-established parameters. The electric pulses cause pores to open on the plasma membrane and facilitate drug transport, enhancing cytotoxicity and reducing side effects. Assessment of electrochemotherapy effects on Ehrlich solid tumor development in this work aims to evaluate in vivo usage of the electroporator device developed by the Department of Electrical Engineering of Engineering School of UFMG. Therefore, 40 Swiss mice were inoculated with Ehrlich tumor cells, and developed the tumor in solid form. After 21 days, mice were subjected to specific treatment protocols (control, bleomycin, electric pulses and electrochemotherapy); 17 days later they were euthanized and the tumors collected for histopathology analysis. Electrochemotherapy induced discrete weight loss and an inflammatory response in the tumor, which was not seen on the other treatment groups. Bleomycin alone induced necrosis. Both groups showed lower cellular proliferation rates. From this study, it was concluded that the animals tolerated electrochemotherapy treatment under anesthesia and the electroporator device developed by the Engineering School of UFMG was adequate when used in an electrochemotherapy protocol.(AU)

Eletroquimioterapia é uma modalidade de tratamento local contra o câncer em que a administração de quimioterápicos não penetrantes à membrana plasmática é associada à aplicação de pulsos elétricos com parâmetros bem estabelecidos, que abrem poros na membrana plasmática e facilitam a entrada desses fármacos nas células, aumentando sua citotoxicidade e reduzindo efeitos colaterais. A avaliação dos efeitos da eletroquimioterapia sobre o desenvolvimento do tumor sólido de Ehrlich em camundongos Swiss neste trabalho teve como objetivo testar o uso in vivo do aparelho eletroporador desenvolvido pelo Departamento de Engenharia Elétrica da Escola de Engenharia da UFMG. Para tanto, foram utilizados 40 camundongos fêmeas da linhagem Swiss, nos quais foram inoculadas células de tumor de Ehrlich, para o desenvolvimento do tumor na forma sólida. Após 21 dias, os camundongos foram submetidos ao protocolo de tratamento específico (controle, bleomicina, pulsos elétricos e eletroquimioterapia); 17 dias depois foram eutanasiados e seus tumores coletados para análise histopatológica e imuno-histoquímica. A eletroquimioterapia induziu perda de peso discreta e uma resposta inflamatória no tumor que não foi observada nos outros grupos. O grupo bleomicina apresentou maior porcentagem de necrose. Ambos os grupos apresentaram menor índice de proliferação celular. Com este estudo, pode-se concluir que o tratamento sob anestesia foi bem tolerado pelos animais e que o aparelho eletroporador desenvolvido pela Escola de Engenharia da UFMG é adequado para utilização em um protocolo de eletroquimioterapia.(AU)

Ciclooxygenase inhibitor and metronomic chemotherapy association for the treatment of metastatic anal sac carcinoma in dog: case report / Inibidor de ciclo-oxigenase associado à quimioterapia metronômica no tratamento de carcinoma de saco anal metastático canino: relato de caso

Metronomic chemotherapy consists of an anticancer modality treatment. It is applicable in patients at an advanced stage, with the objective of increasing overall survival. The aim of this study was to report an anal sac apocrine carcinoma case in a dog with lymph node metastasis treated with metronomic chemotherapy sequential to surgery and conventional chemotherapy using gemcitabine and carboplatin. Metronomic chemotherapy was associated with cyclooxygenase-2 (COX-2) inhibitors, due to strong tumor COX-2 immunohistochemistry expression. Metronomic chemotherapy was initiated with cyclophosphamide, but it was replaced by lomustine, also in metronomic dosage, due to adverse effects. Treatment showed effectiveness, since the patient's overall survival exceeded 1095 days (36 months), considerably higher than the mean overall survival expected for this pathology.(AU)

Quimioterapia metronômica consiste em uma modalidade de tratamento anticancerígeno, aplicável a pacientes em estadiamento avançado, com o objetivo de aumentar a sobrevida global. O objetivo deste trabalho foi relatar um caso de carcinoma apócrino do saco anal, em uma cadela, com metástase em linfonodo tratado com quimioterapia metronômica sequencial à cirurgia e quimioterapia convencional utilizando-se gencitabina e carboplatina. O tratamento metronômico foi associado ao uso de inibidores de ciclo-oxigenase-2 (COX-2), baseando-se na constatação de sua expressão tumoral. A terapia metronômica iniciou-se com ciclofosfamida, mas houve necessidade de substituição pela lomustina, também em dose metronômica, devido à ocorrência de efeitos adversos. O tratamento mostrou ser eficaz, pois a sobrevida do paciente ultrapassa 1095 dias (36 meses) desde a cirurgia, sendo consideravelmente maior que a média relatada para essa patologia.(AU)

Presence of growth hormone receptor (GH-R) mRNA and protein in goat ovarian follicles and improvement of in vitro preantral follicle survival and development with GH.

This study aimed to demonstrate the expression of growth hormone receptor (GH-R) mRNA and protein in goat ovarian follicles in order to investigate the effects of GH on the survival and development of preantral follicles. The ovaries were processed for the isolation of follicles to study GH-R mRNA expression or to localization of GH-R by immunohistochemical analysis. Pieces of ovarian cortex were cultured for 7 days in minimum essential medium(+) (MEM(+)) in the presence or absence of GH at different concentrations (1, 10, 50, 100, and 200 ng/mL). High expression levels of GH-R mRNA were observed in granulosa/theca cells from large antral follicles. However, preantral follicles do not express mRNA for GH-R. Immunohistochemistry demonstrated that the GH-R protein was expressed in the oocytes/granulosa cells of antral follicles, but any protein expression was observed in preantral follicles. The highest (P < 0.05) rate of normal follicles and intermediate follicles was observed after 7 days in MEM(+) plus 10 ng/mL GH (70%). In conclusion, GH-R mRNA and protein are expressed in caprine antral follicles, but not in preantral follicles. Moreover, GH maintains the survival of goat preantral follicles and promotes the development of primordial follicles.

Risk factors associated with Chlamydia psittaci infection in psittacine birds.

Chlamydia psittaci is the aetiological agent of chlamydiosis in birds, especially Psittaciformes. The objective of the present study was to detect C. psittaci by means of semi-nested PCR among psittacine birds sold at pet markets and kept as pet birds in Salvador, Bahia, Brazil. Questionnaires were used to identify risk factors involved in the epidemiology of the disease. In addition, the management of birds and cages was observed at each location studied. The frequency of C. psittaci infection was 10.6% (33/311) in the psittacine birds studied. Birds kept in households were less frequently positive (3.4%; 5/148) than those at pet markets (17.2%; 28/163). Among the several factors analysed in the epidemiology of the disease, only population density (P = 0.001) and cage hygiene (P = 0.041) in birds at pet markets were significantly associated with C. psittaci infection. These results demonstrate the presence of C. psittaci infection in Psittaciformes kept as pets and held at pet markets in Salvador, Bahia, showing that this micro-organism is a public health concern. Control measures should be encouraged to prevent the spread of the agent among birds, as well as among employees and customers.

Photoacoustic investigation of maghemite-based nanocomposite.

Photoacoustic spectroscopy was used to investigate magnetic nanocomposites incorporating nanosized maghemite particles into styrene-divinylbenzene copolymer template. Typical photoacoustic features were observed in bands C, S and L in the wavelength region of 300-1000 nm. The relative intensity of band-C scaled with the nominal concentration of nanosized maghemite incorporated into the polymeric template whereas the lowest relative intensity of band-S was found in the sample in which the template polymerization took place in the presence of the highest polar-like reaction medium. X-ray diffraction and transmission electron microscopy were used to characterize the magnetic nanosized phase as maghemite, with average particle diameter of 6.9 nm (sample Est34), 7.0 nm (sample H30), and 7.9 nm (sample Em15).

Photoacoustic spectroscopy study of Blepharocalyx salicifolius (Kunt) O. Berg.

Photoacoustic spectroscopy (PAS) has revolutionized the fields of biological, environmental, and agricultural sciences. It is a very simple, sensitive, and non-destructive technique that allows the determination of optical properties of bio-samples. The in vivo chlorophylls of the leaf have a recorded maximum absorption peak at 675 nm as against 665 nm of the in vitro chlorophylls. The intensity of purple pigmentation in leaves of Blepharocalyx salicifolius (Kunt) O. Berg, is inversely correlated to the soil moisture levels, leaf water content and leaf water potentials. The applicability of PAS to biological samples was discussed. It allows the validation of existing emission models which are important for atmospheric process. A portable device for photoacoustic spectroscopy of plants and other photosynthetic tissues, cells and organelles is provided. Further, there is provided a method to measure photosynthesis of such tissues, cells and organelles.

Synthesis, characterization and magnetic properties of polymer-Fe3O4 nanocomposite.

The chemical stability of magnetic particles is of great importance for their applications in medicine and biotechnology. The most challenging problem in physics of disordered systems of magnetic nanoparticles is the investigation of their dynamic properties. The chemical coprecipitation process was used to synthesize spherical magnetite nanoparticles of 14 nm. The as-prepared magnetite nanoparticles have been aged in the matrix. Magnetic properties and aging effect were studied by Mössbauer spectroscopy at temperatures ranging from 77 to 300 K, and X-ray diffraction. At room temperature, the Mössbauer spectrum showed superparamagnetic behavior of the particles, while well-defined sextets were observed at 77K, indicating a blocked regime. The superparamagnetic magnetite nanoparticles can be used as microbead biosensors.

Subacute blood pressure behavior in elderly hypertensive women after resistance exercise session.

AIM: The objective of this study was to determine the subacute blood pressure behavior of pharmacologically-treated elderly hypertensive patients after a session of resistance exercise. This was a controlled clinical trial carried out on 30 elderly hypertensive women. The study procedures took place over three days with an interval of 48 hours, and included a test session, an experimental protocol (EP) and a control protocol (CP). METHODS: A 10RM test was carried out to define the EP load for the following exercises: leg press, knee extension, pull-up, biceps curl and machine bench press. In the EP, three sets of exercises with 8 to 10 repetitions were carried out at 2-minute intervals. In the CP all procedures were the same as in the EP, but without the exercises. BP was measured before, immediately after and every ten minutes up to 60 minutes after the EP and CP. RESULTS: There was no significant variation in systolic blood pressure (SBP) or diastolic blood pressure (DBP) in either the EP or the CP immediately after the exercises. In the period up to sixty minutes after the exercises, there was a significant difference in BP, with a decrease in SBP and DBP for both EP and CP. CONCLUSION: The data from the present study offer a good indication that resistance exercises may be prescribed safely to this group of patients. The pressure reduction appears to be influenced by the rest that occurred after the protocols and not by exercise.

Gene expression and immunolocalization of fibroblast growth factor 2 in the ovary and its effect on the in vitro culture of caprine preantral ovarian follicles.

This study quantified Fibroblast growth factor 2 (FGF-2) mRNA and localized FGF-2 protein in different categories of follicles isolated from goat ovaries. In addition, we verified the effects of this factor on the in vitro culture of preantral follicles isolated from goats. For mRNA quantification, we performed real-time PCR using primordial, primary and secondary follicles, as well as cumulus-oocyte complexes (COCs) and mural granulosa and theca cells of small and large antral follicles. For FGF-2 protein localization, the ovaries were subjected to conventional immunohistochemical procedures. Preantral follicles were isolated and cultured in vitro for 12 days in either control (basic) or supplemented with FGF-2 medium. The expression of FGF-2 mRNA was detected in all categories of follicles and there was no difference in preantral follicles and COCs or granulosa/theca cells from small and large antral follicles. However, in large antral follicles, COCs showed expression levels significantly lower than in granulosa/theca cells (p < 0.05). We observed moderate expression of FGF-2 protein in preantral follicles but not in granulosa cells of primordial follicles and theca cells of secondary follicles. In both small and large antral follicles, strong, moderate and weak staining was observed in oocytes, granulosa and theca cells, respectively. The addition of FGF-2 caused a significant increase in the daily follicular growth rate compared to the control group. We conclude that FGF-2 mRNA is expressed throughout follicular development and that its protein can be found in different patterns in preantral and antral follicles. Furthermore, FGF-2 increases the follicular growth rate in vitro.

In vitro inhibition of Streptococci binding to enamel acquired pellicle by plant lectins.

AIM: Initial colonization of the tooth surface by streptococci involves the attachment of these bacteria to adsorbed salivary components of the acquired pellicle. In dental biofilm this adhesion may also involve lectin-like components, present on the surface of the organisms, which bind to complementary carbohydrates on the surface of the tooth. Therefore, this work aimed to evaluate the potential of six lectins, extracted from seeds of Leguminosae family members, to inhibit the adherence of five streptococci species to acquired pellicle in vitro. METHODS AND RESULTS: The lectins used in this work were extracted from Canavalia ensiformis, Canavalia brasiliensis, Dioclea violacea, Dioclea grandiflora, Cratylia floribunda and Vatairea macrocarpa. Fluorescence micrography was employed to visualize the ability of FITC-labeled lectins to attach to acquire pellicle. Adherence inhibition was performed on saliva-coated microtiter plates at which lectins solutions were previously incubated followed by incubation with the oral streptococci. Glucose-mannose specific lectins attached to acquired pellicle with high intensity, while galactose specific lectins, from V. macrocarpa, exhibits low intensity attachment. CONCLUSIONS: All lectins were able to inhibit the adherence of the microorganisms tested (p < 0.01). SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that lectins may be useful in anti adhesion therapeutics.

Ion-exchange chromatography used to isolate a spermadhesin-related protein from domestic goat (Capra hircus) seminal plasma

Mammalian seminal plasma contains among others, proteins called spermadhesins, which are the major proteins of boar and stallion seminal plasma. These proteins appear to be involved in capacitation and sperm-egg interaction. Previously, we reported the presence of a protein related to spermadhesins in goat seminal plasma. In the present study, we have further characterized this protein, and we propose ion-exchange chromatography to isolate this seminal protein. Semen was obtained from four adult Saanen bucks. Seminal plasma was pooled, dialyzed against distilled water and freeze-dried. Lyophilized proteins were loaded onto an ion-exchange chromatography column. Dialyzed-lyophilized proteins from the main peak of DEAE-Sephacel were applied to a C2/C18 column coupled to an RP-HPLC system, and the eluted proteins were lyophilized for electrophoresis. The N-terminal was sequenced and amino acid sequence similarity was determined using CLUSTAL W. Additionally, proteins from DEAE-Sephacel chromatography step were dialyzed and submitted to a heparin-Sepharose high-performance liquid chromatography. Goat seminal plasma after ion-exchange chromatography yielded 6.47 +/- 0.63 mg (mean +/- SEM) of the major retained fraction. The protein was designated BSFP (buck seminal fluid protein). BSFP exhibited N-terminal sequence homology to boar, stallion and bull spermadhesins. BSFP showed no heparin-binding capabilities. These results together with our previous data indicate that goat seminal plasma contains a protein that is structurally related to proteins of the spermadhesin family. Finally, this protein can be efficiently isolated by ion-exchange and reverse-phase chromatography.

Interferon-gamma and tumour necrosis factor-alpha production by CD4+ T and CD8+ T lymphocytes in AIDS patients with tuberculosis.

Tuberculosis (TB) is usually more severe in HIV-infected patients, and the immune derangement found in co-infected patients may differ from that in each isolated disease. Following mitogen stimulation of peripheral blood mononuclear cells (PBMC), interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha production was evaluated in T cells by flow cytometry, and in culture supernatants by enzyme-linked immunosorbent assay (ELISA) in 33 individuals: 11 AIDS patients with tuberculosis, six asymptomatic HIV-1-infected patients, eight patients with tuberculosis and eight healthy controls. The proportion of CD4+ T lymphocytes expressing IFN-gamma did not differ between the groups, whereas a trend towards increased proportions of TNF-alpha-expression in CD4+ T cells was observed in the TB compared to the HIV group, while intermediate values were observed in co-infected patients. Detection of IFN-gamma and TNF-alpha in CD8+ T lymphocytes was higher in TB than in HIV individuals. Co-infected patients presented intermediate values for IFN-gamma, while TNF-alpha detection was similar to that in HIV mono-infection. In conclusion, the proportion of T cells expressing IFN-gamma was relatively preserved in co-infected patients compared to TB patients, while the percentage of T cells expressing TNF-alpha was decreased, mainly in CD8+ T lymphocytes. However, the marked reduction in T lymphocyte numbers in co-infected patients led to a striking reduction of both cytokines in PBMC supernatants, a finding that is consistent with the impaired response to Mycobacterium tuberculosis.

Oral manifestation of sporotrichosis in AIDS patients.

A wide spectrum of oral lesions has been associated with human immunodeficiency viral infection (HIV), or AIDS. This report describes the case of an HIV-infected patient who developed a case of disseminated sporotrichosis whose first clinical sign was the presence of orofacial lesions. A histopathological study of this patient's biopsy specimens taken from the oropharyngeal lesions revealed a number of rounded and/or oval free-spore forms of Sporothrix schenkii, the identification of which was corroborated by culturing skin lesion exudate on Sabouraud's glucose agar. To the best of our knowledge to date, this is the first time a case of the oral manifestation of sporotrichosis in association with HIV infection has been described in the dental literature.

[Gallbladder emptying evaluation in chronic calcifying pancreatitis, by means of a scintilographic study with Tc-99m DISIDA]. / Avaliação do esvaziamento da vesícula biliar em portadores de pancreatite crônica calcificante, através do estudo cintilográfico com Tc-99m DISIDA.

The aim of this study was to analyze the gallbladder motor function in chronic pancreatitis (CP) patients. Gallbladder emptying was evaluated in 11 patients, without and with addition of pancreatic extract and in ten controls. The results were compared and analyzed statistically. The ejection fraction (EF) of the gallbladder (GB) at 30, 45 and 60 minutes were calculated by using Tc-99m DISIDA scintigraphy. The EF of GB at 60 minutes was significantly higher in the controls when compared to patients, although the results between patients were similar without and with addition of pancreatic extract. The results suggest that the delay in the GB emptying does not depend on the eventual alteration in the intestinal phase of the vesicular stimulation, but it probably results from a mechanic factor, which depends on the chronic pathological process located in the head of the pancreas.

[Complications of chronic pancreatitis in São Paulo (Brazil)]. / Complicações das pancreatites crônicas em São Paulo.

Chronic Pancreatitis (CP) presents distinctive characteristics in different geographical areas. With the purpose of evaluating the clinical characteristics, particularly in relation to the frequency and etiopathogeny of the complications in São Paulo, in comparison to other centers, 545 patients with this disease were analyzed, retrospectively, 493 (90.5%) of these patients were males and 52 (9.5%) females, with ages ranging from 8 to 88 (38.2 +/- 9.3 years). The diagnosis of CP was based on criteria previously adopted by the Pancreas Group of Gastroenterology Studies of the Medical School of São Paulo. The principal etiologic factors were represented by: chronic alcoholism in 509 of the 545 patients (93.4%), hereditary factors in four (0.7%), malnutrition in three (0.5%), metabolic alterations in three (0.5%) and obstruction to the pancreatic flow in two patients (0.3%). In 24 (4.4%) of the 545 patients, the etiologic factor could not be established, due to the idiopathic nature of the disease. Of the 509/545 patients (93.4%) presenting chronic ethylism, alcoholic consumption was characterized by: a) mean age alcoholic consumption was initiated: 19.5 +/- 6.5 years; b) mean daily alcoholic consumption in grams of pure ethanol: 358.6 +/- 282.0 g/d; c) mean time of alcoholic consumption: 19.8 +/- 8.8 years, and d) mean age of the appearance of clinical symptoms: 34.9 +/- 9.8 years. The clinical complications were always investigated in accordance with the pre-established protocol; the principal complications observed were represented by: gastrointestinal compression (3.3%), jaundice (24%), cysts (35.9%), cavity effusions (13.3%), pancreatic necrosis (4.7%), abscesses (3.6%), digestive hemorrhage of pancreatic origin (2.3%) and fistulae (1.1%). The incidence of these complications, particularly cysts, cavity effusions and pancreatic necrosis, was greater in the patients studied than in those observed in other regions, probably due to the presence of more intense and frequent anatomopathologic lesions found in these patients, possibly resulting from the greater consumption of alcohol compared to patients with this disease in other centers. The etiopathogenic mechanisms for the complications were individually analyzed and interpreted.

[Giant cell arteritis (Horton's disease): report of 18 cases]. / Arterite de células gigantes (Doença de Horton): a propósito de 18 casos.

The authors present and comment on 18 cases of Giant-cell arteritis observed in an Internal Medicine Department between 1984 and 1991 with emphasis on clinical aspects and diagnostic considerations, on the occult presentation forms and, finally, on two cases with peripheral neuropathy. The clinical manifestations were: headache (78%), general non-specific symptoms (78%), polymyalgia rheumatica (61%), sudden blindness (33%), local temporal signs (28%), jaw pain (24%), articular complaints (17%) and peripheral neuropathy (12%).

[Behcet's disease. Experience of an internal medicine service from 1982 to 1991]. / Doença de Behçet. Experiência de um serviço de medicina interna entre 1982 e 1991.

The authors present the results of a retrospective study on Behçet's disease, concerning 19 patients observed in Medicine Service 2 (Santo António dos Capuchos Hospital) between 1982 and 1991. There were 15 male and 4 female patients, with a mean age of 33. The clinical manifestations were: oral--100%, ocular--100%, genital--89%, articular--79%, cutaneous--68%, general non-specific manifestations--52%, venous--26%, peripheral nervous system--21%, central nervous system--10% and persistent headaches--10%.