The dynamic structure of Spitzenkörpers of Trichosporon asahii examined by the fluorescent probe FM4-64

Abstract The Spitzenkörper is a dynamic and specialized multicomponent cell complex present in the tips of hyphal cells. The amphiphilic styryl dye FM4-64 was found to be ideal for imaging the dynamic changes of the apical vesicle cluster within growing hyphal tips. It is widely used as a marker of endocytosis and to visualize vacuolar membranes. Here we performed uptake experiments using FM4-64 to study the dynamic of the Spitzenkörper in Trichosporon asahii. We observed that Spitzenkörpers were present at the tip of the budding site of the spore, blastospore, and the germ tube of T. asahii. We also found that Spitzenkörpers were present at the tip of the hyphae as well as the subapical regions. Cytochalasin D, an inhibitor of actin polymerization, leads to abnormal Spitzenkörper formation and loss of cell polarity.

Rapid identification of two new isomers in bear bile powder by LC-Q-TOF-MS combined with PCC oxidation / 中国中药杂志

A rapid method of Liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) combined with pyridinium chlorochromate (PCC) oxidation has been developed to determine chemical structures of two novel isomers in bear bile powder. Derivatives of ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) were semi-synthesized by PCC oxidation, then were analyzed by LC-Q-TOF-MS. Separation was carried out on a reverse column with the mobile phase of acetonitrile-0.1% formic acid (45:55). The data of Q-TOF-MS was acquired by MS, MS/MS, positive and negative modes. Since UDCA and CDCA were stereochemical isomeric at an alcohol position, two oxidation products were same and have been confirmed by LC-Q-TOF-MS. Other two products were also determined based on the PCC oxidation theory. Samples of bear bile powder were dissolved by methanol and measured by LC-Q-TOF-MS. Two unknown peaks were found and identified by matching their retention times and accurate mass spectra ions with PCC oxidation productS. Finally, the structures of two new bile acids in bear bile powder were confirmed as 3alpha-hydroxy-7-oxo-5beta-cholanic acid, 7alpha-hydroxy-3-oxo-5beta-cholanic acid, respectively.

Analysis by confocal laser scanning microscopy of the MDPB bactericidal effect on S. mutans biofilm CLSM analysis of MDPB bactericidal effect on biofilm

Since bacteria remain in the dentin following caries removal, restorative materials with antibacterial properties are desirable to help maintaining the residual microorganisms inactive. The adhesive system Clearfil Protect Bond (PB) contains the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) in its primer, which has shown antimicrobial activity. However, its bactericidal effect against biofilm on the dentin has been little investigated. Objective: The aim of this study was to analyze by confocal laser scanning microscopy (CLSM) and viable bacteria counting (CFU) the MDPB bactericidal effect against S. mutans biofilm on the dentin surface. Material and methods: Bovine dentin surfaces were obtained and subjected to S. mutans biofilm formation in BHI broth supplemented with 1% (w/v) sucrose for 18 h. Samples were divided into three groups, according to the primer application (n=3): Clearfil Protect Bond (PB), Clearfil SE Bond, which does not contain MDPB, (SE) and saline (control group). After the biofilm formation, Live/ Dead stain was applied directly to the surface of each sample. Next, 10 µL of each primer were applied on the samples during 590 s for the real-time CLSM analysis. The experiment was conducted in triplicate. The primers and saline were also applied on the other dentin samples during 20, 90, 300 and 590 s (n=9 for each group and period evaluated) and the CFU were assessed by colonies counting. Results: The results of the CLSM showed that with the Se application, although non-viable bacteria were detected at 20 s, there was no increase in their count during 590 s. In contrast, after the PB application there was a gradual increase of non-viable bacteria over 590 s. Conclusions: The quantitative analysis demonstrated a significant decrease of S. mutans CFU at 90 s PB exposure and only after 300 s of Se application. Protect Bond showed an earlier antibacterial effect than Se Bond.

A case of acute hemolytic transfusion reaction due to anti-Di(a) antibody: A case report / 대한마취과학회지

Many medical institutions in Korea have recently been performing an antibody screening test as one of the essential elements of a pre-transfusion test. The Dia antigen is well known as one of the antigens with low incidence among Caucasians; however, it has been discovered with a relatively higher incidence among Mongoloid populations. The frequency of the Dia antigen among the Korean population is estimated to be 6.4-14.5%. But in Korea, a screening panel of cells from abroad without Dia positive cells has been commonly used when a patient has an unexpected antibody screening test. Here we report a case of acute hemolytic transfusion reaction due to Anti-Dia antibody. To prevent other transfusion reaction by anti-Dia antibody, addition of Dia positive cells as unexpected antibody screening test is recommended.

Effect of gamma irradiation on fluoride release and antibacterial activity of resin dental materials

This study evaluated the effect of gamma irradiation on fluoride release and antibacterial activity of FluroShield (FS) and Clearfil Protect Bond (CPB). Four groups were formed: G1-FS + gamma; G2-FS without gamma; G3-CPB + gamma; G4-CPB without gamma. For fluoride release analysis, 12 disks of each material were prepared and covered with nail polish, except for one side (50.4 mm² area). G1 and G3 were sterilized with a 14.5 KGy dose at 27ºC for 24 h, while G2 and G4 (controls) were not sterilized and were maintained under the same time and temperature conditions. Fluoride release measurements were made in duplicate (n=6) by an ion specific electrode. The antibacterial activity of the CPB and FS against Streptococcus mutans after gamma sterilization was evaluated by the agar-disc diffusion method. The diameter of the zones of microbial growth inhibition was recorded after 48 h. Data were analyzed statistically by ANOVA and Tukey's test (α=5 percent). Gamma sterilization decreased the fluoride release of FS by approximately 50 percent, while CPB was not affected. There was no statistically significant difference (p>0.05) in the antibacterial effect of CPB between gamma and non-gamma sterilization groups. FS presented no antibacterial activity. Gamma irradiation decreased the fluoride release of FS, but did not affect the antibacterial activity of the studied materials.

Este estudo avaliou o efeito da esterilização com raios-gama na liberação de flúor e atividade antibacteriana de materiais resinosos, Fluroshield (FS) e Clearfil Protect Bond (CPB). Quatro grupos foram formados: G1-FS e gama; G2-FS sem gama; G3-CPB e gama; G4-CPB sem gama. Doze discos de cada material foram preparados para análise de liberação de flúor, os quais foram cobertos com esmalte de unha, exceto em um lado com 50,4 mm² de área. G1 e G3 foram esterilizados com dose de 14,5 KGy por 24 h/27ºC, enquanto G2 e G4 (controles) não foram esterilizados e foram mantidos sob as mesmas condições de tempo e temperatura. As leituras de liberação de flúor foram feitas em duplicata (n=6) por um eletrodo específico. A atividade antibacteriana foi avaliada pelo teste de difusão em agar. Os halos de inibição foram medidos após 48 h. Os dados foram analisados pelos testes ANOVA e Tukey (α=5 por cento). A esterilização gama diminuiu a liberação de flúor de FS em cerca de 50 por cento, enquanto CPB não foi afetado. Não houve diferença estatisticamente significante entre os grupos esterilizados e controle no efeito antibacteriano do CPB. FS não apresentou atividade antibacteriana. A esterilização gama diminuiu a liberação de flúor de FS, mas não afetou a atividade antibacteriana dos materiais estudados.

Development of Position Encoding Circuit for a Multi-Anode Position Sensitive Photomultiplier Tube / 핵의학분자영상

PURPOSE: The goal of this paper is to present the design and performance of a position encoding circuit for 16 x 16 array of position sensitive multi-anode photomultiplier tube for small animal PET scanners. This circuit which reduces the number of readout channels from 256 to 4 channels is based on a charge division method utilizing a resistor array. MATERIALS AND METHODS: The position encoding circuit was simulated with PSpice before fabrication. The position encoding circuit reads out the signals from H9500 flat panel PMTs (Hamamatsu Photonics K.K., Japan) on which 1.5 x 1.5 x 7.0 mm3 L0.9GSO (Lu1.8Gd0.2SiO5:Ce) crystals were mounted. For coincidence detection, two different PET modules were used. One PET module consisted of a 29 x 29 L0.9GSO crystal layer, and the other PET module two 28 x 28 and 29 x 29 L0.9GSO crystal layers which have relative offsets by half a crystal pitch in x- and y-directions. The crystal mapping algorithm was also developed to identify crystals. RESULTS: Each crystal was clearly visible in flood images. The crystal identification capability was enhanced further by changing the values of resistors near the edge of the resistor array. Energy resolutions of individual crystal were about 11.6%(SD 1.6). The flood images were segmented well with the proposed crystal mapping algorithm. CONCLUSION: The position encoding circuit resulted in a clear separation of crystals and sufficient energy resolutions with H9500 flat-panel PMT and L0.9GSO crystals. This circuit is good enough for use in small animal PET scanners.

Urine products of bone breakdown as markers of bone resorption and clinical usefulness of urinary hydroxyproline: an overview / 中华医学杂志(英文版)

<p><b>PURPOSE</b>The purpose of this study is to review the urine products of bone breakdown as markers of bone resorption and usefulness of urinary hydroxyproline.</p><p><b>DATA</b>Related researches published in 1985 - 2000 were systematically reviewed.</p><p><b>RESULTS</b>Bone markers could be used for early diagnosis of bone metabolic diseases. Biochemical markers of bone resorption that reflect osteoclast activity and/or collagen degradation provide a new and potentially important clinical tool for the assessment and monitoring of bone metabolism. Assessment of bone resorption can be achieved with measurement of urinary hydroxylysine glycosides, urinary excretion of the collagen pyridinium cross-links, urinary excretion of type I collagen telopeptide breakdown products (cross-linked telopeptides) and urinary hydroxyproline.</p><p><b>CONCLUSION</b>Urinary hydroxyproline has been in use as a marker of bone resorption, but it lacks sensitivity and specificity. It is a modified amino acid that is a metabolic product of collagen breakdown. Hydroxyproline may be released either free or with fragments of the collagen molecule attached during bone resorption, and it is also liberated by the breakdown of complement and nonskeletal collagen.</p>

Antidotal efficacy of pyridinium oximes and cholineacetyltransferase inhibitors against organophosphorus intoxication in rodents.

In an attempt to develop effective antidote against organophosphorus intoxication, some new imidazole-pyridinium mono-oximes, long chain pyridinium mono-oximes and cholineacetyltransferase inhibitors were synthesised. These compounds were evaluated for their in vivo therapeutic protection and neuromuscular function studies in rodents. The results indicate that SPK-series oximes may be useful against sarin poisoning without any beneficial effect against VX (O-Ethyl S-2-NN-diisopropylaminoethyl methylphosphonofluoridate) intoxication. The cholineacetyltransferase (ChAT) inhibitors may not be of any help against any of the OP compounds studied in this study.

Cytoarchitectonic arrangement and intracortical circuits of hamster cerebellum. A study by means of confocal scanning laser microscopy

The FM4-64, a member of the family of fluorescent dyes, has been applied to the cerebellar cortex to study its cytoarchitectonic arrangement and intracortical course of mossy and climbing fibers. Slabs of hamster cerebellum, 1-2 mm thick, were incubated in 10, 30 and 100 microM solutions of FM4-64 in 0.1 M sodium phosphate buffer and observed in a slow scan confocal laser scanning microscope. Cellular tomography of cerebellar cortex allowed us to follow the course of mossy and climbing fibers in the cerebellar white and gray substance. They exhibited high fluorescence signal at the level of myelin sheath. Mossy fibers were identified only in the granular layer by their typical rosette formations and dichotomous bifurcation pattern. Climbing fiber bundles were observed crossing the granular layer and giving off collateral branches in the granular and molecular layers in a crossing-over pattern. They ascend to the Purkinje cell layer in their way to the molecular layer. Cerebellar macroneurons (Golgi and Purkinje cells) and microneurons (granule, basket and stellate cells) showed optimal intracellular staining of cell soma, axonal and proximal dendritic processes. The montage of z-series of stacked optodigital sections allowed us to explore in depth the cytoarchitectonic arrangement, nerve and glial cell morphology, and the topographic relationship of intrinsic cerebellar neurons with the afferent mossy and climbing fibers

Use of fluorescent probes to follow membrane traffic in nerve terminals

Optical tracers in conjunction with fluorescence microscopy have become widely used to follow the movement of synaptic vesicles in nerve terminals. The present review discusses the use of these optical methods to understand the regulation of exocytosis and endocytosis of synaptic vesicles. The maintenance of neurotransmission depends on the constant recycling of synaptic vesicles and important insights have been gained by visualization of vesicles with the vital dye FM1-43. A number of questions related to the control of recycling of synaptic vesicles by prolonged stimulation and the role of calcium to control membrane internalization are now being addressed. It is expected that optical monitoring of presynaptic activity coupled to appropriate genetic models will contribute to the understanding of membrane traffic in synaptic terminals.

Serum beta[2]-microglobulin concentration correlates with urinary concentrations of type 1 collagen cross- linked N-telopeptides and deoxypyridinoline in rheumatoid arthritis

Determination of serum BETA[2]-microglobulin concentration, an invasive procedure, has been advocated for monitoring patients' response to treatment in rheumatoid arthritis. The object of this study was to find out if serum BETA[2]-microglobulin concentration correlated with urinary excretions of type 1 collagen cross-linked N-telopeptides [NTx] and deoxypyridinoline [Pyrilinks-D] in rheumatoid arthritis [RA]. Using chemiluminiscent assay, serum BETA[2]-microglobulin concentrations were estimated in 25 female patients with active RA, 25 female with inactive disease, and 25 age-matched healthy female controls. Concentrations of NTx and Pyrilinks-D were also determined by immunoabsorbent assays in spot urine samples from these subject groups. The serum concentration of BETA[2]-microglobulin in patients with RA [7.45 +/- + +/- 2.10 mg/L] was significantly higher [P<0.001] than the concentrations in patients with inactive disease [3.33 +/- 0.76 mg/L], or than in normal healthy controls [2.74 +/- 0.52 mg/L]. Similarly, in patients with active RA, the spot urinary concentrations of NTx [123.08 +/- 25.53 nmol BCE/mmol creatinine] and Pyrilinks-D [15.08 +/- 3.29 nmol/mmol creatinine] were significantly higher [P<0.01] than those in patients with inactive disease [58.42 +/- 12.65 nmol BCE/mmol creatinine and 10.10 +/- 2.43 nmol/mmol creatinine, respectively]. In patients with active RA, serum concentration of BETA[2]-microglobulin correlated positively with spot urinary NTx concentrations [r=0.9910, P=0.0001], and Pyrilinks-D concentration [r=0.6177, P=0.001]. In patients with active RA, the spot urinary concentrations of NTx and Pyrilinks-D correlated positively with serum BETA[2]-microglobulin. Therefore, the estimations of these urinary markers may take the place of serum BETA[2]-microglobulin estimation in monitoring the patient's response to treatment in rheumatoid arthritis

Influência de bochechos com cepacol sobre a placa dentária. Parte I - Avaliação da quantidade de placa formada / Influence of the moutwashes with \"Cepacol\" on the dental plaque

Este estudo demonstrou que bochechos com solução antisséptica contendo cloreto de cetilpiridínio, realizados por adultos jovens, 3 vezes ao dia, proporcionou uma redução de 21,78//nos escores altos de placa e de 31,04//do peso úmido de placa em comparação aos bochechos realizados com uma solução contendo os mesmos ingredientes com exceção do cloreto de cetilpiridínio (placebo). Estes resultados permitem recomendar bochechos com solução antisséptica contendo cloreto de cetilpiridínio, como coadjuvante dos procedimentos mecânicos empregados no controle da placa dentária

Farmacocinética dos bloqueadores neuromusculares / Pharmacokinetics of neuromuscular relaxants

Os autores tecem comentários sobre a farmacocinética dos bloqueadores neuromusculares adespolarizantes e despolarizantes, enfatizando principalmente os aspectos da fisiopatologia que podem alterá-la, como a gravidez, hipotermia, idade avançada e insuficiência renal e hepática

Bacteriemias transitórias por microrganismos anaeróbicos pós-remoçäo de fio de sutura / Transitory bacteriemia due to anerobic microbes after sutura remotion

Visou a constataçäo da ocorrência de hemoculturas positivas, pós-remoçäo do fio de sutura, aposto na mucosa gengival depois de exodontia de molar superior, tendo sido feito, previamente, o uso, como colutório, de uma soluçäo aquosa de cloreto de cetilpiridínio 1:4.000 que embebia a regiäo durante um (1) minuto. Foram utilizados seres humanos do sexo masculino, normo-reativos, os quais foram divididos em grupos: A e B. O grupo A visava a detecçäo de possíveis bacteriemias transitórias, pela simples punçäo venosa do antebraço e a permanência do "butterfly" pelo tempo de duraçäo do teste (cinco minutos). O grupo B se contituía de pacientes que foram submetidos à avulsäo de dentes molares superiores nos quais, após oito dias, removia-se o fio de sutura. Do grupo A colhia-se 3 ml de sangue imediatamente à punçäo venosa e também um minuto e cinco minutos após. Do grupo B colhia-se a mesma quantidade de sangue, pré-remoçäo do fio de sutura e também um minuto e cinco minutos após a remoçäo do mesmo. Todas as hemoculturas foram processadas em meio de cultura para anaeróbicos, conforme as normas preconizadas pela Virginia Polytechnic Institute (VPI). Constatou-se que, em todos os casos do grupo A (dez indivíduos), as hemoculturas foram negativas. Nos pacientes do grupo B (dezoito pacientes), todas as hemoculturas feitas pré-remoçäo do fio de sutura foram negativas e nas hemoculturas realizadas a um minuto e a cinco minutos, após a remoçäo do fio, houve positivância em 50 por cento dos casos. As colônias de microrganismos encontradas foram identificadas quanto ao gênero

Fazadínio em anestesia pediátrica / Fazadimium in pediatric anesthesia

O brometo de fazadínio é um relaxante näo-despolarizante, com início de açäo rápido e sem produzir fasciculaçöes. Apesar da literatura já farta sobre seu emprego em pacientes adultos, os relatos em crianças säo escassos. No presente estudo, quarenta pacientes de 1 a 5 anos foram divididos aleatoriamente em dois grupos de vinte pacientes cada um. A um grupo se administrou brometo de fazadínio e ao outro brometo de pancurônio. Procurou-se comparar a açäo das duas drogas relaxantes através dos seguintes parâmetros: tempos entre a administraçäo das drogas e a parada da atividade respiratória; a qualidade da tubagem traqueal, usando-se o método descrito por Twohig et al. modificado, após um minuto da administraçäo da droga; a pressäo arterial sistólica e diastólica em intervalos de 5, 15 e 30 min após o uso do relaxante e a duraçäo e a forma de reversäo do bloqueio. Todos os pacientes foram intubados pelo mesmo autor, que desconhecia a droga empregada. A avaliaçäo da reversäo do bloqueio foi feita através de um estimulador de nervos periféricos, utilizando-se a seqüência de 4 estímulos. Os resultados obtidos foram tratados estatisticamente, considerando-se o nível de 0,05 (p <0,05) como significativo, para o "t" de Student ou para o "QUI" quadrado. A açäo do fazadínio näo apresentou diferença daquela já conhecida para pacientes adultos. Em doses de 0,4 mg.kg-1 permitiu a tubagem traqueal em 80% dos pacientes em 1`min, com um grau de relaxamento cirúrgico que durou em média 45 min. Apesar de discretamente mais lento no início de açäo, o pancurônio na dose de 0,1 mg.kg-1 se mostrou semelhante ao fazadínio tanto nas condiçöes de tubagem como na duraçäo e recuperaçäo do bloqueio. Ambas as drogas apresentaram uma boa estabilidade cardiocirculatória

Fazadínio: alteraçöes cardiovasculares / Fazadinium: cardiovascular disturbances

O efeito do Fazadínio sobre a pressäo arterial e a freqüência cardíaca foi investigado em 40 pacientes, ASA I e II. Os pacientes foram divididos em dois grupos. O bloqueador neuromuscular foi administrado aos pacientes de cada grupo nas doses de 0,8 e 1,0 mg.k-1, respectivamente. A pressäo arterial foi monitorizada pelo método auscultatório, e a freqüência cardíaca foi determinada por eletrocardiograma. Após o registro dos valores obtidos antes e depois da induçäo com Fentanil (2mcg.kg-1) e Tiopental Sódico (5mg.kg-1) o Fazadínio foi injetado I.V., e novas determinaçöes foram obtidas imediatamente, 2, 5 e 8 minutos após o término da injeçäo. Essa droga produziu aumento da freqüência cardíaca e diminuiçäo da pressäo arterial sistólica e diastólica, sendo o efeito máximo registrado no segundo minuto