Terapia fotodinâmica como adjuvante no tratamento da doença periodontal em equinos / [Adjunctive photodynamic therapy as a treatment for periodontal disease in equines]

A doença periodontal é uma afecção comum, relacionada ao aprisionamento de alimentos em diastemas não fisiológicos, em equinos. O tratamento consiste na correção da causa primária, limpeza e desbridamento do sulco gengival, denominado tratamento convencional (TC). Frequentemente antimicrobianos são necessários, pela gravidade ou patogenicidade dos agentes. A terapia fotodinâmica adjuvante (TF) tem sido estudada pelo seu potencial combate bacteriano, sem causar resistência bacteriana. O objetivo deste estudo foi analisar o uso da TF na doença periodontal, experimentalmente induzida, em dentes incisivos de equinos, e compará-la com o TC. O TC não resultou em melhora clínica estatisticamente, tanto em graus como em profundidade, apenas numérica na profundidade aos 30 dias. A TF foi empregada em dentes com profundidade maior da bolsa periodontal que a do grupo TC e, mesmo assim, apresentou melhora clínica já com sete dias, e mais efetiva aos 30, atingindo, em média, o valor considerado normal, três milímetros. A TF apresentou potencial para ser aplicada na rotina, pelo incremento nos resultados, sem causar efeitos colaterais.(AU)

Periodontal disease is a common condition, related to the entrapment of food in non-physiological diastemas in horses. Treatment consists of correction of the primary cause, cleansing and debridement of the gingival sulcus, called Conventional Therapy (CT). Often antimicrobials are requested due to the severity or pathogenicity of the agents. Adjuvant Photodynamic Therapy (PDT), has been studied for its potential bacterial combat, without causing bacterial resistance. The objective of this study was to analyze the use PDT with the experimentally induced periodontal disease in the incisor teeth of horses, and to compare with the CT. The CT did not result in clinical improvement, either in degrees or millimeters. The PDT was used in teeth with a greater depth of the periodontal pocket than the TC group, and even then, showed clinical improvement in only seven days, and more effective at 30, reaching the three millimeter value considered normal on average. The PDT presented the potential to be applied in the routine by the increase in the results without causing side effects.(AU)

Progesterona plasmática e fertilidade de fêmeas caprinas submetidas à sincronização do estro com prostaglandina F2α / Plasmatic progesterone and fertility of goats submitted at estrus synchronization with Prostaglandin F2α

Avaliou-se a resposta ao protocolo de sincronização do estro com duas doses de prostaglandina F2a (22,5µg) intervaladas de 10 dias, por meio da mensuração da concentração de progesterona plasmática, bem como a taxa de concepção das cabras após a inseminação artificial, de acordo com as diferentes respostas obtidas. Utilizaram-se 23 fêmeas e dois reprodutores da raça Toggenburg. A mensuração da progesterona plasmática foi realizada no dia da primeira aplicação de PGF2a (D0), no D5, no dia da segunda aplicação de PGF2a (D10), no D15, no D20 e no D33. A resposta positiva à PGF2α foi determinada pela queda da concentração de progesterona a valores abaixo de 1,5ng/mL, mensurada nos dias cinco e 15. As fêmeas foram distribuídas em três grupos. O grupo I foi composto por fêmeas que responderam às duas aplicações; o grupo II por fêmeas que não responderam à primeira aplicação e responderam à segunda aplicação; e o grupo III por fêmeas que responderam à primeira aplicação e não responderam à segunda aplicação, foram inseminadas e não conceberam. A presença de um corpo lúteo funcional, no momento das aplicações, determinou a resposta ao protocolo. As diferentes respostas das fêmeas ao protocolo, grupo I e II, não influenciaram as taxas de concepção.

Plasmatic progesterone concentrations were evaluated during the synchronization protocol with two doses of 22.5µg of Prostaglandin F2α, 10-day interval, and the conception rate of females in accordance to the reply to the protocol. Twenty-three female goats and two sexually mature Toggenburg bucks were used. Blood was sampled on day 0 (1st PGF2α injection), and on the following days 5, 10 (2nd PGF2α injection), 15, 20, and 33. The positive reply to the PGF2α was determined when the progesterone concentrations fell to values below 1.5ng/mL at days 5 and 15. The females were divided in three groups: Group I: females that responded to the two PGF2α applications; Group II: females that responded only to the second application; Group III: females that responded the first application, did not respond to the second application, were inseminated but did not conceive. The presence of a functional corpus luteum at the moment of the applications determined the reply to the protocol. There was no difference in the conception rates between females that responded to the two PGF2α applications or responded only to the second application.

Desenvolvimento e caracterização de sistemas semi-sólidos para liberação de própolis na cavidade bucal / Development and characterisation of semisolid systems to deliver propolis in the oral cavity

Pharmaceutical formulations containing poloxamer 407 (P407), Carbopol 934P (C934P) or gelatin (GELA), with ethanolic propolis extract (PE), were designed for the treatment of oral mucosal diseases. PE was produced and its quality was assessed by measuring its specific gravity, pH, weight of dry residue and total flavonoid content. Monopolymeric and binary polymeric formulations were prepared and their gelling temperature (Tsol/gel), pH, continuous flow rheology and mucoadhesion were studied. PE exhibited good quality and the formulations were easy to prepare and showed a wide range of consistency. Most of the formulations showed thermoresponsive behaviour and only those containing 15% P407, plus 0.20% C934P or 1.0 % GELA, displayed Tsol/gel suitable for application to the oral mucosa. Monopolymeric formulations, containing C934P or GELA, and binary formulations exhibited pseudoplastic flow and low degrees of thixotropy. Monopolymeric formulations containing P407 exhibited pseudoplastic flow and rheopexy. The mucoadhesive properties of the systems could not be assessed. Fragments of formulation were found to remain stuck to parts of the mucin disc, owing to cohesive failure of the samples and of the sample/mucin interface. The data obtained on these formulations indicate a potentially useful role in the treatment of oral mucosal diseases

Formulações farmacêuticas contendo poloxamer 407 (P407), Carbopol 934P (C934P) ou gelatina (GELA), e extrato de própolis (EP) foram desenvolvidos para o tratamento de doenças da mucosa oral. EP foi produzido e sua qualidade foi avaliada quanto ao resíduo seco e ao teor de flavonóides totais. Formulações monopoliméricas e poliméricas binárias foram produzidas e a temperatura de gelificação (Tsol/gel), o pH, a reologia, assim como a mucoadesão das mesmas foram avaliados. O EP apresentou boa qualidade, as preparações foram fáceis de produzir e apresentaram uma ampla variação de consistência. A maioria das preparações apresentou comportamento de resposta térmica e apenas as formulações contendo 15% P407 e 0,20% C934P ou 1,0% GELA apresentaram Tsol/gel adequada para a administração na mucosa oral. Formulações monopoliméricas, contendo C934P ou GELA, e binárias exibiram comportamento de fluxo pseudoplástico e baixo grau de tixotropia. Formulações monopoliméricas de P407 exibiram fluxo pseudoplástico e reopexia. As propriedades mucoadesivas dos sistemas não puderam ser avaliadas. Fragmentos de formulações foram encontrados aderidos em alguns lugares do disco de mucina devido à falha de coesão das amostras e da interface amostra/mucina. Os resultados obtidos com essas formulações indicam a utilidade das mesmas no tratamento de doenças da mucosa oral.

Fitoterápicos: um mercado promissor / Phytomedicines: a promising market

Observa-se na Fitoterapia uma tendência de contribuição efetiva à saúde da população. Por consequência, a padronização de fitomedicamentos é um pré-requisito para a garantia da qualidade, bem como para a constância dos efeitos terapêuticos e segurança do usuário. A validação de processo analítico deve garantir, através de evidências experimentais, que o método atenda às exigências das aplicações analíticas, assegurando a confiabilidade dos resultados. Assim, os equipamentos e materiais de laboratório devem ser devidamente calibrados e o analista qualificado. As substâncias químicas de referência devem ser certificadas por compêndios oficiais, como as Farmacopeias ou por outros códigos autorizados pela legislação vigente. Tão importante quanto o desenvolvimento e validação de uma metodologia analítica é o posterior estudo de estabilidade, a fim de garantir que o produto mantenha sua qualidade durante toda vida útil. Para a obtenção de registro de um medicamento fitoterápico dentro dos padrões requeridos pela legislação faz-se necessário, portanto, a realização de diferentes testes para validação deste medicamento de forma a garantir sua segurança no uso, eficácia na utilização e qualidade do produto.

There is an observable trend towards phytotherapy making a recognized effective contribution to public health. Consequently, the standardization of phytomedicines is a prerequisite for quality assurance and to ensure the consistency of therapeutic effects and safety of the user. Analytical method validation should ensure, through experimental testing, that the method meets the requirements of the analytical applications, ensuring the reliability of results. Thus, the equipment and laboratory materials must be properly calibrated and the analyst qualified. The chemical references must be certified by official compendia, such as pharmacopoeias or other officially accepted codes. As important as the development and validation of an analytical methodology is the stability study, performed to ensure that the product retains its quality throughout its shelf-life. Therefore, to register a herbal medicine in accordance with the legal standards, it is necessary to carry out various validation tests on the product, to ensure its safety, effectiveness and quality.

Posttransplant proteinuria associated with everolimus.

Anti-mTOR may induce proteinuria when utilized after renal transplantation. Little is known about the pathogenesis and composition of proteinuria. To clarify this unresolved aspect, we analyzed urinary protein composition utilizing an integrated proteomics approach, including quantitative assays, 2-dimensional electrophoresis, MALDI-TOF, and Western blots among 48 renal transplant recipients treated with everolimus (EVL; n = 31) or enteric-coated mycophenolic acid (EC-MPA; n = 17). High (>3 g/d) or intermediate levels of proteinuria (1-3 g) developed in 12 EVL patients (39%) compared with 4 subjects (23%) in the EC-MPA group. Proteinuria, which started during the first 2 days after EVL, tended to reduce during the follow-up. Quantitative proteomics showed an increase in low molecular proteins beta2 microglobulin (P < .001) and alpha1 microglobulin (P < .025). Qualitative proteomics showed a marked increase among all urinary components in EVL and EC-MPA patients. Major changes involved typical components of glomerular damage: albumin, Zn-alpha1 glycoprotein, alpha2HS glycoprotein, and leucine-rich alpha2 glycoprotein. In addition, we observed specific biomarkers for EVL: clusters of alpha1-antitrypsin fragments and monoclonal lambda chains. In conclusion, EVL induced proteinuria of a mixed glomerular and tubular origin that correlated with the start of treatment and reached nephrotic ranges in few cases. The specific urinary markers may reflect renal alterations related to the transplant or specific alterations associated with the drug.

[18FDG-PET and large vessel vasculitis: preliminary data on 25 patients]. / 18FDG-PET e vasculiti dei grandi vasi: studio preliminare su 25 pazienti.

OBJECTIVE: To evaluate the predictive value of clinical and biochemical features when compared to 18FDG-PET in the diagnostic work-up of large vessel vasculitis (LVV). METHODS: Twenty-five patients underwent 18FDG-PET for the clinical suspect of LVV. All of them presented history of systemic symptoms lasting >or=6 months and laboratoristic evidence of persistently high markers of inflammation. The patients were stratified according with: i) clinical manifestations, defined as presence of one or more ACR criteria for the classification of LVV; ii) laboratory investigations: Erythrocyte Sedimentation Rate (ESR) higher or lower than 50 mm/h, C-Reactive Protein (CRP) higher or lower than 2 mg/dl; iii) prednisone dose in the 4 weeks preceding PET examination. RESULTS: The total number of positive PET was higher in the group without clinical ACR criteria and in the group with inflammation markers under the established cut-off. The number of scans consistent with LVV was higher in the groups presenting one or more clinical criteria for LVV but in those with very high ESR and CRP. In all the cases differences between groups were not statistically significative. A clear cut negative correlation between steroid dose and number of scans suggestive for LVV has been observed. CONCLUSIONS: Diagnosis of LVV remains challenging, especially in patients presenting with a constellation of non-specific symptoms and laboratory findings. In this study, both clinical and biochemical features show low correlation with a vasculitic pattern of FDG uptake. In our experience 18FDG-PET represents an useful diagnostic tool in early stages of LVV and a powerful instrument to follow the treatment responses.

Preparation and characterisation of ethylcellulose microparticles containing propolis

Ethylcellulose microparticles containing propolis ethanolic extract (PE) were prepared by the emulsification and solvent evaporation method. Three ratios of ethylcellulose to PE dry residue value (DR) weretested (1:0.25, 1:4 and 1:10). Moreover, polysorbate 80 was used as emulsifier in the external phase (1.0 or 1.5% w/w). Regular particle morphology without amorphous and/or sticking characteristics was achieved only when an ethylcellulose: DR ratio of 1:0.25 and 1.0% polysorbate 80 were used. Microparticles had a mean diameter of 85.83 mium. The entrapment efficiency forpropolis of the microparticles was 62.99 more or less 0.52%. These ethylcellulose microparticles containing propolis would be useful for developing propolis aqueous dosage forms without the strong and unpleasant taste, aromatic odour and high ethanol concentration of PE.

Recurrent nephrotic syndrome in homozygous truncating NPHS2 mutation is not due to anti-podocin antibodies.

Mutations in NPHS2 are a common cause of focal segmental glomerulosclerosis (FSGS). It was initially assumed that FSGS caused by a genetically defective protein in the native kidney would not recur after transplantation; however, description of three patients with NPHS2 missense mutations challenged the validity of this assumption. A possible mechanism of recurrence in cases with stop-codon mutations is the formation of auto-antibodies against the truncated protein. In this case report, we describe a 9-year-old girl with the R138X NPHS2 mutation who presented with recurrent nephrotic syndrome 4 years after renal transplantation from a deceased donor, and was treated with plasmapheresis with a partial response. Renal histology did not demonstrate glomerular immunoglobulin deposition and an extensive search for anti-podocin antibodies based on indirect Western blot with recombinant podocin, was negative, as was the test for glomerular permeability factor (Palb). Taken together these findings confirm the possibility of post transplantation nephrotic syndrome in patients with NPHS2 mutations. Lack of immunoglobulin deposition, absence of circulating anti-podocin antibodies, and normal Palb suggest that other, unknown pathogenetic mechanisms are implicated.

Glomerular clusterin is associated with PKC-alpha/beta regulation and good outcome of membranous glomerulonephritis in humans.

Mechanisms for human membranous glomerulonephritis (MGN) remain elusive. Most up-to-date concepts still rely on the rat model of Passive Heymann Nephritis that derives from an autoimmune response to glomerular megalin, with complement activation and membrane attack complex assembly. Clusterin has been reported as a megalin ligand in immunodeposits, although its role has not been clarified. We studied renal biopsies of 60 MGN patients by immunohistochemistry utilizing antibodies against clusterin, C5b-9, and phosphorylated-protien kinase C (PKC) isoforms (pPKC). In vitro experiments were performed to investigate the role of clusterin during podocyte damage by MGN serum and define clusterin binding to human podocytes, where megalin is known to be absent. Clusterin, C5b-9, and pPKC-alpha/beta showed highly variable glomerular staining, where high clusterin profiles were inversely correlated to C5b-9 and PKC-alpha/beta expression (P=0.029), and co-localized with the low-density lipoprotein receptor (LDL-R). Glomerular clusterin emerged as the single factor influencing proteinuria at multivariate analysis and was associated with a reduction of proteinuria after a follow-up of 1.5 years (-88.1%, P=0.027). Incubation of podocytes with MGN sera determined strong upregulation of pPKC-alpha/beta that was reverted by pre-incubation with clusterin, serum de-complementation, or protein-A treatment. Preliminary in vitro experiments showed podocyte binding of biotinilated clusterin, co-localization with LDL-R and specific binding inhibition with anti-LDL-R antibodies and with specific ligands. These data suggest a central role for glomerular clusterin in MGN as a modulator of inflammation that potentially influences the clinical outcome. Binding of clusterin to the LDL-R might offer an interpretative key for the pathogenesis of MGN in humans.

Preparation and antimicrobial activity of gelatin microparticles containing propolis against oral pathogens.

Gelatin microparticles containing propolis ethanolic extractive solution were prepared by spray-drying technique. Particles with regular morphology, mean diameter ranging of 2.27 microm to 2.48 microm, and good entrapment efficiency for propolis were obtained. The in vitro antimicrobial activity of microparticles was evaluated against microorganisms of oral importance (Enterococcus faecalis, Streptococcus salivarius, Streptococcus sanguinis, Streptococcus mitis, Streptococcus mutans, Streptococcus sobrinus, Candida albicans, and Lactobacillus casei). The utilized techniques were diffusion in agar and determination of minimum inhibitory concentration. The choice of the method to evaluate the antimicrobial activity of microparticles showed be very important. The microparticles displayed activity against all tested strains of similar way to the propolis, showing greater activity against the strains of E. salivarius, S. sanguinis, S. mitis, and C. albicans.

Circulating anti-actin and anti-ATP synthase antibodies identify a sub-set of patients with idiopathic nephrotic syndrome.

Idiopathic nephrotic syndrome (iNS) with resistance or dependence to steroids is a common disease in children but in spite of an increasing clinical impact its pathogenesis is unknown. We screened for the presence of circulating antibodies against glomerular (podocytes, mesangium) and tubular cells (tubular epithelia) a cohort of 60 children with iNS including 8 patients with a familial trait of iNS or with proven mutation of NPHS1-NPHS2 and 12 with good sensitivity to steroids. Positive sera were found in 8 cases, all belonging to the category without familial trait/molecular defects. The targets of antibodies were characterized with Western blot and MALDI-Mass utilizing beta-hexyl cell extracts separated with two-dimensional electrophoresis. In all cases antibodies of the IgM class were directed against ATP synthase beta chain alone (4 cases) or in combination with actin (3 cases); one child presented IgG against aldose reductase. The clinical picture was nephrotic syndrome with steroid resistance or dependence and variable cyclosporin sensitivity; 3 patients developed end stage renal failure. The basic pathology picture was focal segmental glomerulosclerosis (FSGS) in 4 cases and mesangial proliferative glomerulonephrites with deposition of IgM in 2. Overall, patients with circulating auto-antibodies could not be readely differentiated on clinical grounds with the exception of 3 children who developed positivity for antinuclear antibodies during the follow-up. Affinity-purified IgM from one patient who underwent plasmapheresis for therapeutical pourposes (but not from a normal pool) induced proteinuria in Sprague-Dawley rats and concomitant human IgM deposition within glomeruli. This is the first report of circulating anti-actin/ATP synthase beta chain antibodies in a subset of patients with iNS. Both pathological significance and clinical impact given by the presence of these antibodies and the relationship with other conditions such as lupus-erythematosus, characterized by their presence, must be defined.

Laparoscopic telementored adrenalectomy: the Italian experience.

BACKGROUND: Laparoscopy is widely accepted as the gold standard for adrenalectomy. Telementoring has been developed to reduce the complications associated with surgeon inexperience. We report our preliminary experience with laparoscopic telementored adrenalectomy. METHODS: From July 2002 to May 2003, eight laparoscopic telementored adrenalectomies were performed between two separate operating sites 430 km apart. Six of these procedures were monolateral laparoscopic adrenalectomies, and one was bilateral. All cases were performed by an expert open surgeon who was skilled in laparoscopic procedure but who had no experience in laparascopic adrenalectomy RESULTS: All the procedures were successfully performed in a telementored fashion. The mean operative times, blood loss, and postoperative morbidity results were comparable to those for standard laparoscopic adrenalectomies reported in the literature. CONCLUSIONS: This preliminary experience has demonstrated the feasibility of national telementoring. It is a viable method that can potentially add to surgical education and decrease the likelihood of complications due to inexperience with new techniques.

Limits for the central production of Theta+ and Xi(--)pentaquarks in 920-GeV pA collisions.

We have searched for Theta+(1540) and Xi(--)(1862) pentaquark candidates in proton-induced reactions on C, Ti, and W targets at midrapidity and square root of s = 41.6 GeV. In 2 x 10(8) inelastic events we find no evidence for narrow (sigma approximately 5 MeV) signals in the Theta+ --> pK0(S) and Xi(--) --> Xi- pi- channels; our 95% C.L. upper limits (UL) for the inclusive production cross section times branching fraction B dsigma/dy/(y approximately 0) are (4-16) mub/N for a Theta+ mass between 1521 and 1555 MeV, and 2.5 mub/N for the Xi(--). The UL of the yield ratio of Theta+/Lambda(1520) < (3-12)% is significantly lower than model predictions. Our UL of B Xi(--)/Xi(1530)0 < 4% is at variance with the results that have provided the first evidence for the Xi(--).

In vitro release of propolis from gelatin microparticles prepared by spray-drying technique

The purpose of this study was to investigative the in vitro release of propolis from gelatin microparticles. Gelatin microparticles containing porpolis extractive solution (PES) were prepared by spray-drying technique. Microparticles with a mean diameter of 2.50 µm and with regular morphology were obtained. The entrapment efficiency of propolis in the microparticles was over 39%. Spray-drying showed to be a feasible method for the preparation of gelatin microparticles containing propolis. Comparing to PES, the in vitro release of propolis from gelatin microparticles in aqueous medium was slower, considering markes 1 and 2. Thus, it was possible to transform a liquid propolis dosage form into a solid one, improving manipulation, packaging and storage and with modified release in aqueous medium, comparatively to the ethanolic extract of the drug

Gelatin microparticles containing propolis obtained by spray-drying technique: preparation and characterization.

Gelatin microparticles containing propolis extractive solution (PES) were prepared by spray-drying technique. The optimization of the spray-drying operating conditions and the proportions of gelatin and mannitol were investigated. Regular particle morphology was obtained when mannitol was used, whereas mannitol absence produced a substantial number of coalesced and agglomerated microparticles. Microparticles had a mean diameter of 2.70 microm without mannitol and 2.50 microm with mannitol. The entrapment efficiency for propolis of the microparticles was upto 41% without mannitol and 39% with mannitol. The microencapsulation by spray-drying technique maintained the activity of propolis against Staphylococcus aureus. These gelatin microparticles containing propolis would be useful for developing intermediary or eventual propolis dosage form without the PES' strong and unpleasant taste, aromatic odour, and presence of ethanol.

Hydrogenases in sulfate-reducing bacteria function as chromium reductase.

The ability of sulfate-reducing bacteria (SRB) to reduce chromate VI has been studied for possible application to the decontamination of polluted environments. Metal reduction can be achieved both chemically, by H(2)S produced by the bacteria, and enzymatically, by polyhemic cytochromes c(3). We demonstrate that, in addition to low potential polyheme c-type cytochromes, the ability to reduce chromate is widespread among [Fe], [NiFe], and [NiFeSe] hydrogenases isolated from SRB of the genera Desulfovibrio and Desulfomicrobium. Among them, the [Fe] hydrogenase from Desulfovibrio vulgaris strain Hildenborough reduces Cr(VI) with the highest rate. Both [Fe] and [NiFeSe] enzymes exhibit the same K(m) towards Cr(VI), suggesting that Cr(VI) reduction rates are directly correlated with hydrogen consumption rates. Electron paramagnetic resonance spectroscopy enabled us to probe the oxidation by Cr(VI) of the various metal centers in both [NiFe] and [Fe] hydrogenases. These experiments showed that Cr(VI) is reduced to paramagnetic Cr(III), and revealed inhibition of the enzyme at high Cr(VI) concentrations. The significant decrease of both hydrogenase and Cr(VI)-reductase activities in a mutant lacking [Fe] hydrogenase demonstrated the involvement of this enzyme in Cr(VI) reduction in vivo. Experiments with [3Fe-4S] ferredoxin from Desulfovibrio gigas demonstrated that the low redox [Fe-S] (non-heme iron) clusters are involved in the mechanism of metal reduction by hydrogenases.

Bioremediation of chromate: thermodynamic analysis of the effects of Cr(VI) on sulfate-reducing bacteria.

Developing new bioremediation processes for soils and effluents polluted by Cr(VI) requires the selection of the most efficient and the most heavy-metal-resistant bacteria. The effects of Cr(VI) on bioenergetic metabolism in two sulfate-reducing bacteria (SRB), Desulfovibrio vulgaris Hildenborough and Desulfomicrobium norvegicum, were monitored using isothermal microcalorimetry. The complete reduction of Cr(VI) to Cr(III) was studied by spectrophotometry and by speciation using a combination of high-performance liquid chromatography and inductively coupled plasma-mass spectrometry. Results revealed that Cr(VI) induces an inhibition of growth with concomitant production of energy, which can be compared to the reaction of the bacteria to a stress such as oxidative stress. Moreover, the sensitivity of bacteria towards this metal is as a characteristic of the strain, which leads to differences in the kinetics of Cr(VI) reduction. The study by microcalorimetry of heavy metal effects on SRB bioenergetic metabolism thus appears an appropriate tool to identify better strains to be used for industrial bioremediation process development.

Barkas effect for antiproton stopping in H2.

We report the stopping power of molecular hydrogen for antiprotons of kinetic energy above the maximum (approximately 100 keV) with the purpose of comparing with the proton one. Our result is consistent with a positive difference in antiproton-proton stopping powers above approximately 250 keV and with a maximum difference between the stopping powers of 21%+/-3% at around 600 keV.

Cytochromes c555 from the hyperthermophilic bacterium Aquifex aeolicus. 2. Heterologous production of soluble cytochrome c555s and investigation of the role of methionine residues.

The cycB2 gene encoding the soluble cytochrome c555s from Aquifex aeolicus, an hyperthermophilic organism, has been cloned and expressed using Escherichia coli as the host organism. The cytochrome was successfully produced in the periplasm of an E. coli strain coexpressing the ccmABCDEFGH genes involved in the cytochrome c maturation process. Comparison of native and recombinant cytochrome c555s shows that both proteins are indistinguishable in terms of spectroscopic and physicochemical properties. Since two different methionine residues are present in the sequence stretch usually providing the sixth ligand to the heme iron, site-directed mutagenesis has been performed in order to identify the methionine serving as the axial ligand. Two single mutations were introduced, leading to the replacement of each methionine by a histidine residue. Characterization of both mutants, M78H and M84H cytochromes c555s, using biochemical and biophysical techniques has been carried out. The M84H mutant exhibits spectral features identical to those of native cytochrome. Its redox midpoint potential is decreased by 40 mV. By contrast, substitution of methionine 78 by a histidine residue strongly alters the structural and physicochemical properties of the molecule which exhibits characteristics of His/His iron coordination type rather than His/Met. These results allow us to identify methionine 78 as the sixth ligand of cytochrome c555s heme iron. Preliminary results on the thermostability of the native and mutant cytochromes c555 are also reported.

Inhibition of renal permeability towards albumin: a new function of apolipoproteins with possible pathogenetic relevance in focal glomerulosclerosis.

Focal segmental glomerulosclerosis (FSGS) is a degenerative renal disease characterized by the accumulation of extracellular matrix and lipids within the glomerular tuft. It has been proposed that an abnormal renal permeabilization towards proteins induced by a putative plasma factor is, in some way, involved in the pathogenesis of the disease. In this paper, we measured the plasma permeability activity (Palb) in several sera of patients with FSGS and found a mean activity of 0.82+/-0.03 which means a marked increase compared to a mean Palb of 0.16+/-0.03 in normal controls. Coincubation of FSGS and normal serum reduced the permeability activity within the normal range; normal serum added to the incubation medium after the glomeruli had already been exposed to the FSGS serum had no effect, suggesting the presence of inhibitory substances with a direct effect on a circulating substrate. Finally, the antipermeability activity was retained when heated to 60 degrees C but not to 100 degrees C. By serial fractionations of normal serum and reported activity measurements at each step, five natural occurring inhibitors of albumin permeabilization were purified and characterized by matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS), as components of apolipoproteins (apo) (apo E2 and E4, apo L, the high Mr apo J and a 28 kDa fragment of apo A-IV). Coincubation of each apolipoprotein with FSGS serum inhibited permeability, but only apo J and apo E2 and E4 were found to be crucial for the process. In conclusion, we have purified from normal serum five inhibitors of permeability induced by FSGS serum, all corresponding to apolipoproteins. An imbalance between permeability factors and apolipoproteins may play a pathogenetic role in FSGS.