Fungal infection control by garlic extracts (Allium sativum L) and modulation of peritoneal macrophages activity in murine model of sporotrichosis / Controle da infecção fúngica pelos extratos de alho (Allium sativum L) e modulação da atividade de macrófagos peritoneais em modelo murino de esporotricose

Abstract Garlic (Allium sativum L.) is grown all over the world as seasoning and medicinal vegetable since 3,000 BC. Allicin is the main component of garlic, being attributed to it the most of its biological activities, such as bactericidal, antifungal and antiviral actions. However, other compounds of garlic present antioxidant, hypocholesterolemic, vasodilator activities, protective action against different types of cancer, and immunomodulatory. Fungal infections are important causes of morbidity and mortality in people mainly in immunosuppressed ones. Sporothrix schenckii, the causing agent of Sporotrichosis (most common subcutaneous mycosis in Latin America), is dimorphic fungus, of saprophytic life in soil or plants, infecting people and animals mainly through skin injuries and bruises. The main of this work was to evaluate the influence of garlic consuming on immune modulation of healthy and infected Swiss mice in induced way by S. schenckii, since these animals functioning of peritoneal macrophages as well as the nitric oxide and cytokines' production (IL-1β, IL-10 and IL-12) and to evaluate the antifungal potential of garlic with S. schenckii through minimum inhibitory concentration test and colony-forming units. The results showed that garlic offers antifungal potential with S. schenckii. The oral taking of garlic extracts influences the releasing of cytokines by macrophages, regular consuming shows anti-inflammatory effect, and its acute use may take to an inflammatory response. Mice that consumed garlic responded more effectively to fight against the infection.

Resumo O alho (Allium sativum L.) é cultivado em todo o mundo como hortaliça condimentar e medicinal desde 3.000 a. C. A alicina é o principal componente do alho, sendo atribuída a ela a maior parte das suas atividades biológicas, dentre elas as ações bactericida, antifúngica e antiviral. Porém, outros compostos do alho apresentam atividade antioxidante, hipocolesterolemiante, vasodilatadora, ação protetora contra diversos tipos de câncer e imunomoduladora. As infecções por fungos são causas importantes de morbidade e mortalidade no homem principalmente em indivíduos imunossuprimidos. O Sporothrix schenckii, agente causal da esporotricose (micose subcutânea mais comum na América Latina), é fungo dimórfico, de vida saprofítica no solo ou em vegetais, infectando homens e os animais principalmente através de lesões e arranhões na pele. O objetivo desse trabalho foi avaliar a influência do consumo de alho na imunomodulação de camundongos Swiss saudáveis e infectados de forma induzida por S. schenckii, a partir do estado funcional dos macrófagos peritoneais desses animais quanto à produção de óxido nítrico e das citocinas (IL-1β, IL-10 e IL-12) e avaliar o potencial antifúngico do alho frente ao S. schenckii por meio de teste de concentração inibitória mínima e unidades formadoras de colônia. Os resultados demonstraram que o alho apresenta potencial antifúngico frente S. schenckii. A administração oral de extratos de alho influencia a liberação de citocinas por macrófagos, o consumo regular apresenta efeito anti-inflamatório, e seu uso agudo pode gerar uma resposta inflamatória. Camundongos que consumiram alho responderam de forma mais efetiva no combate da infecção.

Fungal infection control by garlic extracts (Allium sativum L.) and modulation of peritoneal macrophages activity in murine model of sporotrichosis.

Garlic (Allium sativum L.) is grown all over the world as seasoning and medicinal vegetable since 3,000 BC. Allicin is the main component of garlic, being attributed to it the most of its biological activities, such as bactericidal, antifungal and antiviral actions. However, other compounds of garlic present antioxidant, hypocholesterolemic, vasodilator activities, protective action against different types of cancer, and immunomodulatory. Fungal infections are important causes of morbidity and mortality in people mainly in immunosuppressed ones. Sporothrix schenckii, the causing agent of Sporotrichosis (most common subcutaneous mycosis in Latin America), is dimorphic fungus, of saprophytic life in soil or plants, infecting people and animals mainly through skin injuries and bruises. The main of this work was to evaluate the influence of garlic consuming on immune modulation of healthy and infected Swiss mice in induced way by S. schenckii, since these animals functioning of peritoneal macrophages as well as the nitric oxide and cytokines' production (IL-1ß, IL-10 and IL-12) and to evaluate the antifungal potential of garlic with S. schenckii through minimum inhibitory concentration test and colony-forming units. The results showed that garlic offers antifungal potential with S. schenckii. The oral taking of garlic extracts influences the releasing of cytokines by macrophages, regular consuming shows anti-inflammatory effect, and its acute use may take to an inflammatory response. Mice that consumed garlic responded more effectively to fight against the infection.

Measurements of gamma dose and thermal neutron fluence in phantoms exposed to a BNCT epithermal beam with TLD-700.

Gamma dose and thermal neutron fluence in a phantom exposed to an epithermal neutron beam for boron neutron capture therapy (BNCT) can be measured by means of a single thermoluminescence dosemeter (TLD-700). The method exploits the shape of the glow curve (GC) and requires the gamma-calibration GC (to obtain gamma dose) and the thermal-neutron-calibration GC (to obtain neutron fluence). The method is applicable for BNCT dosimetry in case of epithermal neutron beams from a reactor because, in most irradiation configurations, thermal neutrons give a not negligible contribution to the TLD-700 GC. The thermal neutron calibration is not simple, because of the impossibility of having thermal neutron fields without gamma contamination, but a calibration method is here proposed, strictly bound to the method itself of dose separation.

Methods for dose measurements in small phantoms irradiated at BNCT epithermal column.

Suitable dosimeter methods have been proposed and tested, to measure the different dose contributions in small phantoms exposed to epithermal/thermal neutron beams designed for BNCT. One method is based on Fricke-gel dosimeter in small tubes of 2.8mm of external diameter, that allow determining profiles of gamma dose and of boron dose. The other method is based on the use of TLD-700 chips, from whose answer the contribution of thermal neutrons is subtracted by means of appropriate parameters of the glow curve.

Neutron fluence depth profiles in water phantom on epithermal beam of LVR-15 research reactor.

Horizontal channel with epithermal neutron beam at the LVR-15 research reactor is used mainly for boron neutron capture therapy. Neutron fluence depth profiles in a water phantom characterise beam properties. The neutron fluence (approximated by reaction rates) depth profiles were measured with six different types of activation detectors. The profiles were determined for thermal, epithermal and fast neutrons.

LVR-15 reactor epithermal neutron beam parameters--results of measurements.

The epithermal neutron beam of the LVR-15 reactor provides the appropriate conditions for varied BNCT activity. The principal parameters have been frequently determined. The following detectors have been used for the measurement: set of activation monitors of different nuclides irradiated in free beam and in the water phantom, Si semiconductor detector with (6)LiF converter, twin ionization chambers, thermoluminescence dosimeters, gel dosimeters used for imaging of separate part of dose, neutron spectrometer of Bonner type. Obtained results of measured parameters are presented in the paper.

Dose imaging in a thorax phantom with lung-equivalent volume at the epithermal neutron beam of LVR-15 reactor.

A thorax phantom has been designed, consisting of PMMA and PE plates containing a cavity filled with a laboratory-made lung-substitute. Fricke-gel dosimeters have been placed in the lung-substitute volume, and the phantom has been irradiated at the epithermal column of LVR-15 reactor. Absorbed dose images have been obtained for both gamma radiation and charged particles emitted in the (10)B reactions with thermal neutrons. Measurements with thermoluminescence dosimeters (TLDs) and Monte Carlo (MC) calculations have been performed too, in order to attain inter-comparison of results.

Evaluation of all dose components in the LVR-15 reactor epithermal neutron beam using Fricke gel dosimeter layers.

Fricke gel dosimeters in the form of layers are suitable to reconstruct bidimensional distributions of the absorbed dose; in accordance with their chemical composition and applying suitably developed algorithms, they can provide dose images of the different radiation components in a BNCT field. After the description of the applied method, this work presents the results obtained at the epithermal column of the BNCT facility at the NRI in Rez (CZ). The measured dose distributions are shown in comparison with data taken by means of other dosimeters thermoluminescence dosimeters (TLDs) and with calculations carried out with the Monte Carlo code MCNP5. The agreement with the results obtained by means of the different techniques is satisfying.

Development of equipments for determination of BNCT source spectral parameters.

The knowledge of neutron and gamma ray energy spectra can strongly influence the BNCT information about delivered dose to target volume as well as to the surface healthy tissue region. This region is very often decisive to stay within the recommended healthy tissue limit. Modification of neutron Bonner spectrometer to one block i.e. Bonner spectrometer monoblock (BSM) and gamma ray Si semiconductor spectrometer are being developed and verified in real conditions of LVR-15 reactor beam. Test measurements were also carried out in conditions of known standard spectra. The accepted procedure and the first results documenting the sensitivity BSM to different spectra are presented.

[Neutron capture therapy in the treatment of glioblastoma multiforme. Initial experience in the Czech Republic]. / Neutronová záchytová terapie v lécbe glioblastoma multiforme. První zkusenosti v Ceské republice.

BACKGROUND: Glioblastoma multiforme is the most frequent primary brain tumor in adults. Despite advances in surgery, radiotherapy and chemotherapy, its treatment remains unsatisfactory with very limited overall survival. In the year 2001, in cooperation with Department of Neurosurgery, Nemocnice Na Homolce and Nuclear Research Institute in Rez, we have started to treat glioblastoma patients with boron neutron capture therapy (BNCT). METHODS AND RESULTS: Cells of malignant brain tumors, especially that of glioblastomas, are able to accumulate boron compounds. If BNCT should be successful, it is necessary to reach selective accumulation of sufficient amount of 10B in the tumor and low accumulation in the normal brain tissue. After BSH administration, radiation with low energy thermal neutrons is delivered. It results in nuclear capture and fission reactions with subsequent selective damage of tumor cells. At the time of analysis 9 patients have been enrolled. Therapy was completed in 5 patients. Treatment has been very well tolerated. We observed minimal acute toxicity associated with radiation and no laboratory abnormalities after administrations of BSH. Unfortunately treatment results were quite unsatisfactory. The median time to progression and overall survival were shorter then expected with conventional treatment. CONCLUSIONS: BNCT is very well tolerated with only a modest toxicity. In contrast to standard radiation, BNCT patients receive only one dose of radiation. Nevertheless, in this small pilot study first results were inferior when compared either to outcomes of conventional therapy or to results reported from other BNCT groups. It might be explained that lower dose of radiation had been used. Further study will show whether the higher dose radiation can improve treatment results.

Determinants of recombinant production of antimicrobial cationic peptides and creation of peptide variants in bacteria.

Cationic peptides possessing antibacterial activity are virtually ubiquitous in nature, and offer exciting prospects as new therapeutic agents. We had previously demonstrated that such peptides could be produced by fusion protein technology in bacteria and several carrier proteins had been tested as fusion partners including glutathione-S-transferase, S. aureus protein A, IgG binding protein and P. aeruginosa outer membrane protein OprF. However these fusion partners, while successfully employed in peptide expression, were not optimized for high level production of cationic peptides (Piers, K., Brow, M. L., and Hancock, R. E. W. 1993, Gene 137, 7-13). In this paper we took advantage of a small replication protein RepA from E. coli and used its truncated version to construct fusion partners. The minimal elements required for high level expression of cationic peptide were defined as a DNA sequence encoding a fusion protein comprising, from the N-terminus, a 68 amino acid carrier region, an anionic prepro domain, a single methionine and the peptide of interest. The 68 amino acid carrier region was a block of three polypeptides consisting of a truncated RepA, a synthetic cellulose binding domain and a hexa histidine domain. The improved system showed high level expression and simplified downstream purification. The active peptide could be yielded by CNBr cleavage of the fusion protein. This novel vector was used to express three classes of cationic peptides including the alpha-helical peptide CEMA, the looped peptide bactenecin and the extended peptide indolicidin. In addition, mutagenesis of the peptide gene to produce peptide variants of CEMA and indolicidin using the improved vector system was shown to be successful.

Neutron-capture therapy of brain tumours: neutron sources, neutron-capture drugs, biological tests and clinical perspectives in the Czech Republic.

The paper reviews neutron sources, chemical compounds and clinical perspectives of the boron neutron-capture therapy of brain tumours. Special attention is paid to the physical characteristics and biological effectiveness of the epithermal neutron beam constructed at the LVR-15 nuclear reactor at Rez near Prague.

Monitoring of the boron neutron-capture reaction by a simple animal model.

Sodium borocaptate (BSH, Na2Bl2HllSH), a slow neutron-capture compound, was injected into the left forebrain ventricle of 1-week-old rats (150 microg BSH/3 microl phosphate buffered saline). After 90 min, the animals were irradiated by epithermal neutrons (LVR-15 nuclear reactor in Rez near Prague, flux density 8.8 x 10(7) neutrons cm-2 s-1, 8 MW reactor power, 8.2 cGy/min) for 5, 10 or 20 min. The brains were examined histologically 8 h after irradiation. In animals irradiated for 5 to 10 min (41 and 82 cGy-Eq, respectively) lethal damage of cells was found in the external granular layer of the cerebellum and the subependymal layer of the forebrain. Irradiation for 20 min (164 cGy-Eq) caused more extensive destruction of cell populations in these regions and, in addition, dead cells appeared also in the more differentiated postmitotic compartments, namely the deeper layers of the cerebellum, layers II/III of the cerebral cortex and corpus callosum. In the forebrain periventricular layer, the extent of cell damage was declining towards the olfactory bulbs. In intact animals, as well as in those injected only with the 150 microl phosphate buffered saline, the radiation damage was low and limited only to the most sensitive dividing populations of the cerebellum and the forebrain. The study demonstrates a differentiation-dependent damage of the rat brain cells by alpha particles and presents a simple model for evaluation of the biological effectiveness of slow neutron beams constructed for neutron-capture therapy of tumors.

A genotoxicological study of hexachlorobenzene and pentachloroanisole.

The potential mutagenic activity of hexachlorobenzene (HCB) and pentachloroanisole (PCA) was investigated. No genotoxicity after application on Salmonella typhimurium (Ames test), Escherichia coli, and human peripheral blood lymphocytes in vitro was observed.

Anorexia nervosa / Anorexia nervosa

Se analiza el caso de un nino con anorexia nervosa y se discute tanto el diagnostico diferencial como la dinamica familiar que explica el problema

[Even distribution of myocardial protection in surgery of the coronary arteries: successive cardioplegia at the origin of the aorta and at the coronary sinus]. / Pour une répartition uniforme de la protection myocardique dans la chirurgie des coronaires: cardioplégie successive dans la racine de l'aorte et dans le sinus coronaire.

In coronary bypass surgery myocardial protection by injection of a cardioplegic fluid is made uneven by the presence of stenotic segments. In order to make it even, the authors suggest that the fluid should be injected first into the aortic origin, then into the coronary sinus. This method should provide greater safety when multiples bypasses are performed and in cases of impaired left ventricular function.