Influence of in vitro capacitation time on structural and functional human sperm parameters / 亚洲男科学杂志(英文版)

A cascade of dramatic physiological events is linked to the sperm acrosome reaction and binding to the oocyte's zona pellucida during human sperm capacitation. However, structural and functional sperm changes during capacitation currently remain poorly defined. Here, we performed a multibiomarker approach based on the utilization of sperm concentration, motility, viability, morphology, acrosome reaction, tyrosine phosphorylation, DNA fragmentation, and lectin-binding sites to analyze the impact caused by swim-up selection times (uncapacitated, 1 h capacitated, and 4 h capacitated) on sperm function and structure in normozoospermic samples. We found that a 4 h swim-up capacitation increased sperm quality, because a large number of cells with normal morphology and lower DNA fragmentation rates were recovered. Furthermore, the long-term capacitation induced a higher percentage of cells with tyrosine phosphorylation of the principal piece as well as a redistribution of lectin-binding sites. Overall, the multivariate biomarkers analyzed showed a less variable distribution on spermatozoa recovered after 4 h capacitation than that with the shorter capacitation time. These findings stress the importance of capacitation time as a relevant factor in sperm quality with potential biological reproductive implications both for basic research and in assisted reproduction techniques.

Antibacterial mechanisms of ethyl acetate extract from endophytic fungi Alternaria alternata against Staphylococcus aureus / 中草药

Objective To investigate the effect of ethyl acetate extract (B06e) from fermentation liquid of an endophytic fungus Alternaria spp. on the cell membrane integrity and the permeability of Staphylococcus aureus. Methods The minimum inhibitory concentration (MIC) of B06e against S. aureus was measured by double dilution method; The changes of electric conductivity of bacterial culture, A260 and A280 before and after treated by B06e were analyzed, respectively. Besides, the changes of cell membrane permeability before and after by B06e was measured by flow cytometry. The effect of B06e on the cell membrane structure was investigated by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR). Results The results showed that MIC value of B06e against S. aureus was 50 μg/mL. The conductivity of 3 × MIC treatment group was 1.06 times of the value of the control group; after treatment of B06e, the values of A260 and A280 were significantly higher than those of the control group: The beta-galactosidase activity of 3 × MIC treatment group was 9.43 times more than the value of the control group; Flow cytometry analysis showed that 3 × MIC treatment group by propidium iodide (PI) staining of positive cells was 47.63 times more than the control group; SEM and FT-IR analysis showed that the structure of bacterial cell changed after B06e treatment. Conclusion B06e can kill S. aureus cell by increasing the permeability of its cell membrane and destroy cell membrane integrity.

In Vitro Antifungal Activity of (1)-N-2-Methoxybenzyl-1,10-phenanthrolinium Bromide against Candida albicans and Its Effects on Membrane Integrity

Metal-based drugs, such as 1,10-phenanthroline, have demonstrated anticancer, antifungal and antiplasmodium activities. One of the 1,10-phenanthroline derivatives compounds (1)-N-2-methoxybenzyl-1,10-phenanthrolinium bromide (FEN), which has been demonstrated an inhibitory effect on the growth of Candida spp. This study aimed to explore the in vitro antifungal activity of FEN and its effect on the membrane integrity of Candida albicans. The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of FEN against planktonic C. albicans cells were determined using the broth microdilution method according to the Clinical and Laboratory Standards Institute guidelines. Cell membrane integrity was determined with the propidium iodide assay using a flow cytometer and were visualized using scanning electron microscopy (SEM). Planktonic cells growth of C. albicans were inhibited by FEN, with an MIC of 0.39–1.56 µg/mL and a MFC that ranged from 3.125 to 100 µg/mL. When C. albicans was exposed to FEN, the uptake of propidium iodide was increased, which indicated that membrane disruption is the probable mode of action of this compound. There was cells surface changes of C. albicans when observed under SEM.

Black tea extract protects against -radiation-induced membrane damage of human erythrocytes.

The membrane integrity of circulating red blood cells (RBCs) is compromised by the deleterious actions of -radiation in humans. Tea is the most widely consumed popular, inexpensive and non-toxic beverage rich in antioxidants. Here, we explored the radioprotective actions of black tea against the -radiation-induced membrane permeability of human erythrocytes. The phytochemical analysis of tea revealed the total polyphenol content to be 114.89±6.03 mg gallic acid equivalent/g dry wt. and flavonoid content, 34±0.11 mg catechin equivalent/g dry wt. of the extractable solid in the commercially available tea bags. Tea extracts showed potential scavenging of H2O2 and NO, appreciable extent of total antioxidant capacity and effective anti-hemolytic action. Tea extracts (15 µg/mL) significantly ameliorated the -radiation-induced increase of the levels of thiobarbituric acid-reactive substances (TBARS, an index of lipid peroxidation) in the RBC membrane ghosts. Stored blood showed higher levels of K+ ion as compared to the normal blood which was elevated by -radiation. Membrane ATPase was inhibited by the exposure to -radiation. Treatment of RBCs with the tea extracts (15 µg/ml) prior to the exposure of -radiation significantly mitigated these changes in the erythrocyte membranes caused by the lower dose of radiation (4 Gy) as compared to that induced by the higher dose of -radiation.

Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation / Identificação de danos ultraestruturais e funcionais em espermatozoides de tatus-peba (Euphractus sexcinctus) oriundos da criopreservação

The aim of the study was to cryopreserve the semen of six-banded armadillos (Euphractus sexcinctus) in Tris-yolk and glycerol diluent, and to determine the damage caused by the freezing-thawing process, using fluorescent markers and ultrastructural analysis. Semen samples (n=11) collected from 4 adult six-banded armadillos by electroejaculation were cryopreserved in Tris diluent plus 20% egg yolk and 3% glycerol, in a fast freezing curve. Classical analysis of samples was performed after dilution, refrigeration and thawing, followed by fluorescence analysis, using a combination of fluorescent probes to assess membrane integrity (propidium iodide - PI and Hoechst - H342), and mitochondrial activity (CMXRos - Mito Tracker Red®). We also used the ultrastructural analysis to verify possible morphological alterations caused by cryoinjuries. When compared with fresh samples, we verified a significant decline in all the armadillos' semen parameters after thawing, in which only 6.1% motile sperm were found. However, the percentage of sperm which remained with viable (13%) and functional (24.7%) membranes after thawing suggests that some cells could be live but immotile. Analysis using fluorescent markers revealed that the mitochondria of armadillos' sperm is highly sensible to the freezing protocol and the findings through ultrastructure analysis proved this statement. Additionally, the images obtained by transmission electron microscopy revealed that frozen-thawed sperm presented damaged plasma membrane, nuclear modifications as changes in chromatin and acrossomal changes relative to sperm capacitation. In conclusion, this study is the first attempt to cryopreserve the semen of an armadillo species, and to help us to identify critical points on the freezing-thawing procedure in order to improve the protocol.(AU)

O objetivo deste estudo foi criopreservar o sêmen de tatus-peba (Euphractus sexcinctus) em diluente Tris-gema e glicerol, e determinar os danos causados pelo processo de congelação-descongelação, utilizando marcadores fluorescentes e análise ultraestrutural. As amostras de sêmen (n=11) coletadas de 4 tatus-peba adultos por eletroejaculação foram criopreservadas em diluente Tris acrescido de 20% de gema de ovo e 3% de glicerol, em curva rápida de congelação. A análise clássica das amostras foi realizada após a diluição, refrigeração e descongelação, seguida por análise de fluorescência, utilizando uma combinação de sondas fluorescentes para avaliar a integridade da membrana (Iodeto de Propídio - PI e Hoechst - H342), e a atividade mitocondrial (CMXRos - Mito Tracker RED®). Foi também utilizada a análise ultraestrutural para verificar possíveis alterações morfológicas causadas pela crioinjúria. Quando comparadas com as amostras a fresco, verificou-se uma queda significativa em todos os parâmetros seminais dos tatus após a descongelação, em que apenas 6,1% de espermatozoides móveis foram encontrados. No entanto, o percentual de espermatozoides que permaneceu com membrana viável (13%) e funcional (24,7%) após a descongelação sugere que algumas células podem estar vivas, mas imóveis. Análises utilizando marcadores fluorescentes revelaram que as mitocôndrias dos espermatozoides de tatus são altamente sensíveis ao protocolo de congelação e os achados através da análise ultraestrutural comprovaram esta afirmação. Além disso, as imagens obtidas por microscopia eletrônica de transmissão revelaram que espermatozoides congelados-descongelados apresentaram membranas plasmáticas danificadas, modificações nucleares como alterações na cromatina, e alterações acrossomais relativas à capacitação espermática. Em conclusão, este estudo é a primeira tentativa de criopreservação de sêmen em uma espécie de tatu, e nos auxiliou a identificar pontos críticos no processo de congelação-descongelação, a fim de melhorar o protocolo.(AU)

Bacillus cereus as a nongastrointestinal pathogen.

The potential of Bacillus cereus to cause systemic infections is of serious concern. Apart from Gastrointestinal infections, it causes respiratory tract infections, nosocomial infections, eye infections, CNS infections, cutaneous infections, endocarditis, osteomyelitis and urinary tract infections. The potential of this bacterium to cause life threatening infections has increased. Trauma is an important predisposing factor for Bacillus cereus infections. The maintenance of skin and mucous membrane integrity limits infection by this micro-organism.

Improved assessment of frozen/thawed mouse spermatozoa using fluorescence microscopy

Genetically modified (GM) animals are unique mutants with an enormous scientific potential. Cryopreservation of pre-implantation embryos or spermatozoa is a common approach for protecting these lines from being lost or to store them in a repository. A mutant line can be taken out of a breeding nucleus only if sufficient numbers of samples with an appropriate level of quality are cryopreserved. The quality of different donors within the same mouse line might be heterogeneous and the cryopreservation procedure might also be error-prone. However, only limited amounts of material are available for analysis. To improve the monitoring of frozen/thawed spermatozoa, commonly used in vitro fertilization (IVF) followed by embryo transfer were replaced with animal-free techniques. Major factors for assessing spermatozoa quality (i.e., density, viability, motility, and morphology) were evaluated by fluorescence microscopy. For this, a live/dead cell staining protocol requiring only small amounts of material was created. Membrane integrity was then examined as major parameter closely correlated with successful IVF. These complex analyses allow us to monitor frozen/thawed spermatozoa from GM mice using a relatively simple staining procedure. This approach leads to a reduction of animal experiments and contributes to the 3R principles (replacement, reduction and refinement of animal experiments).

Cysteine addition on short-term cooled boar semen preservation and its relationship with swine field fertility / Adição de cisteína na preservação de sêmen suíno refrigerado e sua relação com a fertilidade suína

Artificial insemination is routinely used in the swine industry to reduce the costs of production through to increase the efficiency of the refrigerated boar semen process. The objective of this study was to evaluate the effect of different levels of cysteine (CYS) added to the Beltsville Thawing Solution (BTS) extender semen during cooling for up to 72 hours. Ejaculated from three boars were collected with the gloved-hand technique and semen aliquots were diluted in BTS as follow: BTS only (BTS), BTS + 0.1mM cysteine (CYS0.1), BTS + 0.5mM cysteine (CYS0.5), BTS + 1.0mM cysteine (CYS1.0), BTS + 2.5mM cysteine (CYS2.5), BTS + 5.0mM cysteine (CYS5.0), BTS + 10.0mM cysteine (CYS10.0), and BTS + 20.0mM cysteine (CYS20.0). Evaluation of sperm integrity were analyzed using 0.5mg/ml propidium iodide (plasma membrane), 100µg/ml isothiocynate-conjugated Pisum sativun agglutinin (acrosomal membrane) and 153µM 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide (mitochondria potential) after semen dilution at specific times (0, 24, 48 and 72 hours). Additionally, we also evaluated the effects of 5.0 mM CYS addition in the BTS extender on the maintenance of sperm quality and their influence on fertility in the swine production. After artificial insemination, animals were evaluated based on the estrous return and the number of piglet's born. Cysteine at concentrations of 10.0 and 20.0mM resulted in more pronounced reductions even at the time zero. Semen viability decreased to levels below 10 percent at these high levels of CYS in the first 24 hour of storage at 17ºC. At the end of the storage time, less than 65 percent of sperm cells had intact plasma membrane in all groups. The sperm viability decreased significantly when the semen was added at high concentrations of CYS (time "0"; CYS10.0 and CYS20.0; p<0.05), when compared to the other CYS concentrations. The BTS (10.20±0.39) treated group showed a lower rate of estrus return when compared to other (BTSCYS; 86.05±039), and it showed also the highest total number of piglets borne per treatment (12.71±3.38 vs. 9.00±3.38, respectively). In conclusion, the addition of CYS in the BTS semen extender did not maintain spermatic viability of boar cooled spermatozoa and it results in a higher percentage of return to estrus and lower number of piglets borne.

A inseminação artificial é usada rotineiramente na indústria suinícula para reduzir os custos de produção além de obter maior eficiência reprodutiva durante o processo de resfriamento do sêmen. O objetivo deste trabalho foi avaliar o efeito da adição de diferentes concentrações de cisteína (CIS) ao diluidor de sêmen Beltsville Thawing Solution (BTS) resfriado sobre a qualidade espermática por até 72 horas. Foram coletados ejaculados de três cachaços e as amostras de sêmen foram diluídas em BTS, conforme os seguintes tratamentos: BTS (grupo controle); CIS0,1 (BTS + 0,1mM de cisteína); CIS0,5 (BTS + 0,5mM de cisteína); CIS1,0 (BTS + 1,0mM de cisteína); CIS2,5 (BTS + 2,5mM de cisteína); CIS5,0 (BTS + 5,0mM de cisteína); CIS10,0 (BTS + 10,0mM de cisteína) e CIS20,0 (BTS + 20,0mM de cisteína). A avaliação da integridade espermática foi determinada através de sondas fluorescentes em uma combinação de 100µg/mL FICT-PSA (isotiocinato de lecitina), 0.5mg/ml PI (iodeto de propidio), e 153µM JC-1 (5,5',6,6'-tetracloro-1,1',3,3'-tetraetillbenzimidazolil iodeto de carbocianina). As avaliações dos tratamentos foram realizadas 0, 24, 48 e 72 horas após a diluição do sêmen. Adicionalmente, foi avaliado o efeito da adição de 5,0 mM de cisteína ao diluidor BTS na manutenção da qualidade espermática e no efeito na fertilidade em suínos. Após a inseminação artificial, as fêmeas foram avaliadas quanto a taxa de retorno e o tamanho da leitegada. Durante todos os períodos analizados, os grupos CIS10,0 e CIS20,0 apresentaram menor número de espermatozóides viáveis em relação aos demais grupos. A viabilidade espermática diminuiu a níveis abaixo de 10 por cento nos tratamentos CIS10,0 e CIS20,0 nas primeiras 24 horas de armazenamento a 17ºC. Ao final do período de armazenamento todos os grupos apresentavam média inferior a 65 por cento de espermatozóides com a membrana plasmática intacta. A viabilidade espermática diminuiu significativamente quando altas concentrações de CIS (hora "0"; CIS10,0 e CIS20,0; p<0.05) foram adicionadas ao sêmen comparadas com as demais concentrações. O grupo BTS (10,20±0,39) apresentou menor taxa de retorno ao estro comparado com BTSCIS (86,05±0,39), além de apresentar maior número de leitões nascidos (12,71±3,38 vs . 9,00±3,38, respectivamente). Portanto, podemos concluir que a adição de CIS ao diluidor BTS não mantém a qualidade espermática e resulta em maior taxa de retorno ao estro e menor número de leitões nascidos.

Efecto de la centrifugación sobre la membrana plasmática y el ADN de espermatozoides bovinos / Efeito da centrifugação sobre a membrana plasmática e o DNA de espermatozóides bovinos / Effect of centrifugation on plasma membrane and DNA of bovine spermatozoa

El objetivo de este estudio fue determinar si la integridad de la membrana plasmática y el ADN de los espermatozoides pueden ser afectados por la centrifugación realizada en el proceso de lavado y selección. Los espermatozoides fueron sometidos a diferentes tiempos de centrifugación (10, 30 y 45 min); se utilizó un control negativo con espermatozoides no centrifugados y un control positivo con espermatozoides sometidos a estrés oxidativo con peroxido de hidrógeno (H2O2) (200 μM). Para evaluar la integridad de la membrana se utilizó la prueba hipoosmótica (HOST) y para evaluar la fragmentación del ADN se utilizó el ensayo cometa: El análisis estadístico se realizó mediante la prueba de Fisher. La centrifugación durante 10, 30 y 45 min, presentó un efecto estadísticamente significativo sobre el daño en el ADN, respecto de los espermatozoides no centrifugados (p<0.05). Además, se observó diferencia estadística significa (p<0.05) entre los espermatozoides centrifugados a diferentes tiempos con respecto al control positivo realizado con H2O2 (200 μM). La comparación de medias no indicó diferencia estadística significativa entre los espermatozoides no centrifugados y los centrifugados por un periodo de 10 y 30 min (p>0.05) en la reacción positiva a la prueba HOST, lo cual sí sucedió (p<0.05) al comparar los no centrifugados y los centrifugados por 45 min. El control positivo realizado con H2O2 presentó diferencia significativa (p<0.05) con el resto de los tratamientos. En conclusión, los resultados del presente trabajo sugieren que la centrifugación de los espermatozoides durante 10, 30 ó 45 min a 700 x g para la realización del gradiente diferencial de Percoll, tiene un efecto deletéreo sobre el ADN de los espermatozoides bovinos y que la centrifugación por 45 min además de causar daño en el ADN produce pérdida de la integridad de la membrana plasmática.

O objetivo de este estudo foi determinar se a integridade da membrana plasmática e o DNA dos espermatozóides podem ser afetados pela centrifugação realizada no processo de lavado e seleção. Os espermatozóides foram submetidos a diferentes tempos de centrifugação (10, 30 e 45 min); foi utilizado um controle negativo com espermatozóides não centrifugados e um controle positivo com espermatozóides submetidos a estresse oxidativo com peróxido de hidrogênio (H2O2) (200 μM). Para avaliar a integridade da membrana foi utilizado o teste hipoosmótica (HOST) e para avaliar a fragmentação do DNA Fo utilizado o ensaio cometa: A análise estatística se realizou mediante o teste de Fisher. A centrifugação durante 10, 30 e 45 min, apresentou um efeito estatisticamente significativo sobre o dano no DNA, em relação aos espermatozóides não centrifugados (p<0.05). Além disto, observou-se diferença estatisticamente significativa (p<0.05) entre os espermatozóides centrifugados a diferentes tempos em relação ao controle positivo realizado com H2O2 (200 μM). A comparação de medias não detectou diferença estatisticamente significativa entre os espermatozóides não centrifugados e os centrifugados por um período de 10 e 30 min (p>0.05) na reação positiva à prova HOST, o qual ocorreu ao comparar os não centrifugados e os centrifugados por 45 min (p<0,05). O controle positivo realizado com H2O2 apresentou diferença significativa (p<0.05) quando comparado contra os outros tratamentos. Em conclusão, os resultados do presente trabalho sugerem que a centrifugação dos espermatozóides durante 10, 30 ou 45 min a 700 x g para realização do gradiente diferencial de percoll, têm um efeito deletério sobre o DNA dos espermatozóides bovinos e que a centrifugação por 45 min além de causar dano no DNA produz perda da integridade da membrana plasmática.

The aim of this study was to evaluate the effects of different times of centrifugation on plasma membrane integrity and DNA of bovine sperm cells, by means of the hyposmotic test (HOST) and the comet assay, respectively. The sperm cells were centrifuged at 700 x g for 10, 30 or 45 min. No-centrifuged thawed semen served as negative control whereas hydrogen peroxide (H2O2) (200 mM) treated sperm cells were used as a positive control. The results indicate that while the integrity of the plasma membrane was not affected by centrifugation, bovine sperm DNA was damaged independently of centrifugation times. Significant differences between negative control and treatments were found (p<0.05) and in the same way, between treatments and positive control, where the damage for oxidative stress was greater. These results indicate that centrifugation could be detrimental for in vitro bovine embryo production. Additionally, some grade of DNA fragmentation in not centrifuged sperm cells (negative control) was registered, suggesting that DNA of bovine sperm cells could be affected by other factors, probably freezing procedure.

Initial tissue response of biodegradable membrane in rat subcutaneous model / 대한치주과학회지

PURPOSE: Various kinds of biodegradable membranes are currently used in dental clinics. And the frequency and the necessity of their usage are increasing due to their numerous advantages. Therefore it is important to understand the difference of various membranes and histological reaction against implanted membranes. MATERIALS AND METHODS: Biodegradable membranes of Biogide(R), Resolute(R), and Tutodent(R) were cut into small pieces by 1.0x0.5cm. The membranes were implanted 1.5cm apart from each other under the epithelium on the skull of 18 Sprague Dawley rats. The animals were sacrificed at 3, 7, and 14 days after surgical procedure. The specimens were examined by histological analysis. RESULTS: 1. Early period after implantation of the membranes showed connective tissues surrounding membranes and there were a few inflammatory cells present. 2. In Biogide(R) and Tutodent(R) specimens, inflammatory cells and surrounding tissues were shown to infiltrate from outside with slight density difference inside. In Resolute(R) specimens, membranes were fragmented. Inflammatory cells and connective tissues were also observed inside. 3. In Resolute(R) specimen, giant cells were present which implicates that foreign body reaction has occurred. 4. Biogide(R) had lower integrity than other membranes and is not enough to be used alone in defect area. However, Resolute(R) had superior firmness than others. Tutodent(R) had middle level of integrity. CONCLUSION: This experimental model enabled to observe early inflammatory reactions and morphological changes of materials and can be used to develop and evaluate the efficacy of biodegradable membranes. Duplication of standardized human oral environment will be required in future experiments.

Viability evaluation of engineered tissues

Biohybrid artificial organs encompass all devices capable of substituting for an organ or tissue function and are fabricated from both synthetic materials and living cells. The viability of engineered tissue could be related to the viability of implanted cells. The system of viability assay for mammalian cell culture can be applied to the determination of cell viability for engineered tissue. This review explores various methods of cell viability assay which can be applied to the viability evaluation of engineered tissue. The major criteria employed in viability assays include survival and growth in tissue culture, functional assay, metabolite incorporation, structural altercation, and membrane integrity. Each viability assay method is based on different definitions of cell viability, and has inherent advantages and disadvantages. In order to be able to assess the viability of cells with one assay method, it is desirable to compare the viability measurements from various assays derived from different criteria.

Contractile Reserve versus Cell Membrane Integrity for Predicting Contractile Recovery after Reperfusion in Acute Myocardial Infarction and Their Relationship

BACKGROUND: Early prediction of functional recovery could have very important prognostic and therapeutic implication. Modalities evaluating functional recovery of dysynergic segments have included metabolic and perfusion imaging as well as assessment of contractile reserve in response to inotropic agent. Dobutamine stress echocardiography(DSE) assesses contractile reserve whereas rest and 24hr delayed Tl-201 SPECT(T1-201) assesses cell membrane integrity. Does contractile reserve always imply cell membrane integrity? If not, which one could be more useful predictor of contractile recovery of the infarcted segment after restoration of the infarct related artery(IRA) in acute myocardial infarction(AMI)? The aim of this study was to evaluate comparative accuracy of contractile reserve and cell membrane integrity in predicting contractile recovery and their relation. METHODS: We prospectively compared DSE with Tl-201 in 22 of 34 patients with acute myocardial infarction from August 1996 to September 1997. AU patients underwent coronary angiography and got revascularization treatment if they had significant stenosis. DSE and Tl-201 were done within 24hrs after successful restoration without flow limiting residual stenosis. An intravenous infusion of dobutamine(5(mg/kg/min) was started with an infusion pump and continued in 3-minute dose increment up to 20(mg/kg/min under continuous ECG and echocardiographic monitoring. Regional wall motion was assessed according to 16-segment model from American Society of Echocardiography recommendation. Wall motion was scored as 1 (normal), 2(mild to moderate hypokinesia), 3(severe hypokinesia), 4(akinesia), 5(dyskinesia). The improvement of wall motion score(WMS) more than 1 from asynergy during dobutmine infusion was considered to have contractile reserve. SPECT images were obtained at rest and 24hours later. The Tl-201 images were analyzed visually according to 16 segment model, scored by grade 0: normal uptake, 1: mild to moderate decreased, 2: severe decreased, 3: defect. Those segments were considered to have a cellular integrity if the defect at rest improved more than grade 1 on delayed images. All patients underwent 1 month follow up echocardiography after discharge. The improvement of WMS more than score 1 from asynergy at follow up was considered to have contractile recovery. RESULTS: 71 akinetic segments out of total of 352 segments were analyzed. There was no significant difference of sensitivity(88 vs 82%), specificity(82 vs 82%), and accuracy(86 vs 81%) in predicting contractile recovery between DSE and Tl-201. The agreement between contractile reserve and cellular integrity was 73% with kappa value of 0.42(p=0.001). CONCLUSION: Contractile reserve has similar sensitivity, specificity and predictive value with cell membrane integrity in predicting contractile recovery. However contractile reserve immediately after restoration does not necessarily imply cell membrane integrity in acute myocardial inFarction.

Effect of griseofulvin on lipid composition and membrane integrity in Microsporum gypseum.

The effect of griseofulvin on lipid constituents and membrane permeability of Microsporum gypseum has been investigated. Mycelia grown in medium containing griseofulvin (IC50 concentration) possessed a lower content of total lipids, phospholipids and sterols. This inhibitory effect was further supported by decreased incorporation of [14C] acetate in total lipids, total phospholipids and sterols. Decrease in total phospholipids was also reflected to a varying extent in all individual phospholipids. An increase in the unsaturated to saturated fatty acid ratio was observed in mycelia grown in medium containing griseofulvin. Membrane permeability was affected by griseofulvin as shown by increased K+-efflux and greater leakage of intracellular [32P] labelled components from prelabelled cells. Our results suggest that the antifungal activity of griseofulvin is partially due to its secondary effect on lipid constituents of Microsporum gypseum.