Yeast infection of sperm, oocytes and embryos after intravaginal culture for embryo transfer

We report two patients treated with interferon-alpha who developed Raynaud's phenomenon followed by multiple digital necrosis. Arteriography of the legs revealed diffuse distal narrowing. Histological examination of the resected tissue showed ischaemic necrosis within the dermis and subcutaneous tissue without vasculitis. Resolution of the symptoms was observed when interferon-alpha was withdrawn. Since other causes of Raynaud's phenomenon and digital necrosis were ruled out, a drug cause is likely. Raynaud's phenomenon should be looked for in patients treated with interferon-alpha. If present, immediate withdrawal of the drug is required.     

Partitioning of the Golgi apparatus during mitosis in living HeLa cells

The Golgi apparatus of HeLa cells was fluorescently tagged with a green fluorescent protein (GFP), localized by attachment to the NH2-terminal retention signal of N-acetylglucosaminyltransferase I (NAGT I). The location was confirmed by immunogold and immunofluorescence microscopy using a variety of Golgi markers. The behavior of the fluorescent Golgi marker was observed in fixed and living mitotic cells using confocal microscopy. By metaphase, cells contained a constant number of Golgi fragments dispersed throughout the cytoplasm. Conventional and cryoimmunoelectron microscopy showed that the NAGT I-GFP chimera (NAGFP)-positive fragments were tubulo-vesicular mitotic Golgi clusters. Mitotic conversion of Golgi stacks into mitotic clusters had surprisingly little effect on the polarity of Golgi membrane markers at the level of fluorescence microscopy. In living cells, there was little self-directed movement of the clusters in the period from metaphase to early telophase. In late telophase, the Golgi ribbon began to be reformed by a dynamic process of congregation and tubulation of the newly inherited Golgi fragments. The accuracy of partitioning the NAGFP-tagged Golgi was found to exceed that expected for a stochastic partitioning process. The results provide direct evidence for mitotic clusters as the unit of partitioning and suggest that precise regulation of the number, position, and compartmentation of mitotic membranes is a critical feature for the ordered inheritance of the Golgi apparatus.     

Lymphotactin: a key regulator of lymphocyte trafficking during acute graft rejection

One strategy for improving the antitumor selectivity and toxicity profile of antitumor agents is to design drug carrier systems with suitable transport proteins. Thus, four maleimide derivatives of doxorubicin were bound to thiolated human serum albumin which differed in the stability of the chemical link between drug and spacer. In the maleimide derivatives, 3-maleimidobenzoic or 4-maleimidophenylacetic acid was bound to the 3'-amino position of doxorubicin through a benzoyl or phenylacetyl amide bond and 3-maleimidobenzoic acid hydrazide or 4-maleimidophenylacetic acid hydrazide was bound to the 13-keto position through a benzoyl hydrazone or phenylacetyl hydrazone bond. The acid-sensitive albumin conjugates prepared with the carboxylic hydrazone doxorubicin derivatives exhibited an inhibitory efficacy in the MDA-MB-468 breast cancer cell line and U937 leukemia cell line comparable with that of the free drug (using the BrdU-(5-bromo-2'-deoxyuridine)-incorporation assay and tritiated thymidine incorporation assay respectively, IC50 approximately 0.1-1 microM) whereas conjugates with the amide derivatives showed no or only marginal activity. These results demonstrate that antiproliferative activity depends on the nature of the chemical bond between doxorubicin and carrier protein. Acid-sensitive albumin conjugates are suitable candidates for further in vitro and in vivo assessment.     

Differential expression and functions of cortical myosin IIA and IIB isotypes during meiotic maturation, fertilization, and mitosis in mouse oocytes and embryos

To explore the role of nonmuscle myosin II isoforms during mouse gametogenesis, fertilization, and early development, localization and microinjection studies were performed using monospecific antibodies to myosin IIA and IIB isotypes. Each myosin II antibody recognizes a 205-kDa protein in oocytes, but not mature sperm. Myosin IIA and IIB demonstrate differential expression during meiotic maturation and following fertilization: only the IIA isoform detects metaphase spindles or accumulates in the mitotic cleavage furrow. In the unfertilized oocyte, both myosin isoforms are polarized in the cortex directly overlying the metaphase-arrested second meiotic spindle. Cortical polarization is altered after spindle disassembly with Colcemid: the scattered meiotic chromosomes initiate myosin IIA and microfilament assemble in the vicinity of each chromosome mass. During sperm incorporation, both myosin II isotypes concentrate in the second polar body cleavage furrow and the sperm incorporation cone. In functional experiments, the microinjection of myosin IIA antibody disrupts meiotic maturation to metaphase II arrest, probably through depletion of spindle-associated myosin IIA protein and antibody binding to chromosome surfaces. Conversely, the microinjection of myosin IIB antibody blocks microfilament-directed chromosome scattering in Colcemid-treated mature oocytes, suggesting a role in mediating chromosome-cortical actomyosin interactions. Neither myosin II antibody, alone or coinjected, blocks second polar body formation, in vitro fertilization, or cytokinesis. Finally, microinjection of a nonphosphorylatable 20-kDa regulatory myosin light chain specifically blocks sperm incorporation cone disassembly and impedes cell cycle progression, suggesting that interference with myosin II phosphorylation influences fertilization. Thus, conventional myosins break cortical symmetry in oocytes by participating in eccentric meiotic spindle positioning, sperm incorporation cone dynamics, and cytokinesis. Although murine sperm do not express myosin II, different myosin II isotypes may have distinct roles during early embryonic development.     

Comparison of the isoacceptor makeup of transfer RNA for 2 early stages in the development of loach embryos

The chromatographic profiles of isoacceptor tRNA's for 9 amino acids from the loach embryos at two early developmental stages were compared by the method of chromatography on sepharose column in the decreasing gradient of ammonium sulphate concentration to elucidate the possible role of tRNA in the control of early embryogenesis processes. Certain differences both in the proportions of some isoacceptor peaks and their relative positions on the chromatographic profile were found for 8 aminoacyl tRNA. Methionyl-tRNA was represented by one constant peak.     

Perturbation of the mitosis/apoptosis balance: a fundamental mechanism in toxicology

BACKGROUND: The greatest risk of psychiatric morbidity during the climacteric years is linked to a greater exposure to stressful life experiences, women's dissatisfaction with their role in society and to an absence of social backup. The question to be posed now is the extent to which the longitudinal surveys can confirm, refute or complement the results of the cross-sectional analyses. METHODS: A study involving population-based cohorts was carried out on a sample of 120 women who had been previously identified as being premenopausal or menopausal during a cross-sectional examination conducted between 1987 and 1988 within the city of Valencia (Spain). The aims were to analyze the movements of specific psychosocial factors (role satisfaction, level of social support) making a comparison between the premenopausal and postmenopausal phases. RESULTS: An increase in the frequency of psychiatric episodes was detected in the later stages of the climacteric. The absence of global social support, being a possible psychiatric case and experiencing severe life events during the initial phases of the climacteric all emerge as being the most reliable factors for predicting the psychiatric morbidity during the postmenopausal phase. CONCLUSIONS: The results lend weight to the hypothesis of psychiatric morbidity being linked to social changes. A discussion of the clinical implications is presented.     

Change in fibrinolytic activity as a parameter for assessing local mechanical stimulation during physical exercise

The influences of mechanical force during physical exercise on blood coagulation and fibrinolysis were investigated and the effects of sports equipment, especially shoes were also examined. As an experimental model, local mechanical vibration was applied to the palm of the hand. General coagulation parameters did not change, but fibrinolytic activity was elevated due to the tissue plasminogen activator (t-PA) released from vascular endothelial cells stimulated by vibration. The influence of mechanical stimulation by repeated side-jumping with bare feet was examined on the sole of the foot. As with t-PA, the von Willebrand Factor antigen (vWF:Ag) derived from vascular endothelial cells also tended to increase, but not as much as when wearing sports shoes. Sports shoes protected the blood vessels of the feet from damage by mechanical force during physical exercise. Changes in fibrinolytic activity and t-PA could be useful for assessing local mechanical stimulation during physical exercise and also as indexes for the development of new sports equipment and its improvement.     

Localization of the 26S proteasome during mitosis and meiosis in fission yeast

The 26S proteasome is a large multisubunit complex involved in degrading both cytoplasmic and nuclear proteins. We have investigated the localization of this complex in the fission yeast, Schizosaccharomyces pombe. Immunofluorescence microscopy shows a striking localization pattern whereby the proteasome is found predominantly at the nuclear periphery, both in interphase and throughout mitosis. Electron microscopic analysis revealed a concentration of label near the inner side of the nuclear envelope. The localization of green fluorescent protein (GFP)-tagged 26S proteasomes was analyzed in live cells during mitosis and meiosis. Throughout mitosis the proteasome remained predominantly at the nuclear periphery. During meiosis the proteasome was found to undergo dramatic changes in its localization. Throughout the first meiotic division, the signal is more dispersed over the nucleus. During meiosis II, there was a dramatic re-localization, and the signal became restricted to the area between the separating DNA until the end of meiosis when the signal dispersed before returning to the nuclear periphery during spore formation. These findings strongly imply that the nuclear periphery is a major site of protein degradation in fission yeast both in interphase and throughout mitosis. Furthermore they raise interesting questions as to the spatial organization of protein degradation during meiosis.     

Managed care in a new key: Britain's strategies for the 1990s

The new Labour government in Britain has issued three variations of a White Paper that outline significant changes in how the world's largest managed care health system will be run. All three emphasize systemwide criteria for quality, effectiveness, and health gain, which in turn imply redressing past inequalities in funding and service. One, the Scottish Paper, eliminates Thatcher's internal market and returns to a simple organizational structure centering on the health boards. The other two propose combining primary care practices into purchasing groups, a daunting task that will spawn many new problems and expenses as it recreates the internal market.     

Ureteral opening pressure: a novel parameter for the evaluation of pediatric hydronephrosis

PURPOSE: The antegrade nephrostogram is an important tool in the evaluation of the upper urinary tract. However, the information currently provided by a nephrostogram is largely limited to anatomical details. To establish a meaningful pressure-flow parameter that may be incorporated into a routine nephrostogram, we evaluated the ureteral opening pressure (defined as the pressure at which contrast material is first seen beyond the suspected site of obstruction) and correlated these findings with the results of pressure-flow studies performed with an external infusion and/or furosemide induced diuresis. MATERIALS AND METHODS: A total of 52 renal units were studied under a prospective pressure-flow study protocol. All patients had grade 3 or 4 hydronephrosis (Society of Fetal Urology classification) and patient age range was 0.2 to 12 years (median 1.1). The suspected sites of obstruction were the ureteropelvic and ureterovesical junctions in 42 and 10 renal units, respectively. With the patient under general anesthesia 22 gauge percutaneous nephrostomy needles were inserted. Pressure-flow studies with an external infusion and/or furosemide induced diuresis were then performed. As the renal pelvic pressure progressively increased during the course of the pressure-flow studies, the renal pelvic pressure at which contrast material was first seen to appear distal to the suspected site of obstruction was recorded as the ureteral opening pressure. Ureteral opening pressures were compared to the results of the pressure-flow studies. RESULTS: With a positive test defined as renal pelvic pressure greater than 14 cm. water, positive ureteral opening pressures were associated with positive pressure-flow study results in 100% of the cases, regardless of which form of pressure-flow study was used or where the suspected site of obstruction was located. In contrast, negative ureteral opening pressures had specificities and negative predictive values of only 19 to 57%, depending on the form of the pressure-flow study and the suspected site of obstruction. CONCLUSIONS: An elevated ureteral opening pressure was 100% predictive of obstruction and may obviate the need for more elaborate pressure-flow analyses. However, if the ureteral pelvic pressure remained low, the possibility of a potentially significant obstruction could not be definitively eliminated and further evaluation was required.     

Influence of rat placental lactogen-I on the development of whole rat embryos in culture

Rat placental lactogen-I (rPL-I), the first prolactin-like hormone expressed in the placenta during pregnancy in the rat, is known to influence maternal functions. In the present study, we have investigated the effects of rPL-I on the growth and development of cultured whole rat embryos. Rat embryos, with or without ectoplacental cone (EPC) attached, were explanted at day 9 of gestation. After 48 h of culture, the embryos, enclosed by the yolk sacs, were assessed by the presence of visible heart contractions ('heart beats'), crown-rump length (CRL) and yolk sac diameter (YSD). When intact embryos with EPC were cultured, the concentrations of rPL-I and rPL-II (products of EPC) in the medium were 850+/-841 and 92+/-181 ng/ml respectively (means+/-s.e.m.). In embryo cultures with the EPC removed, rPL-I levels decreased to相似文献   

The ovarian hyperstimulation syndrome in extracorporeal oocyte fertilization and the transfer of cleavage-stage embryos

Major clinical characteristics of ovarian hyperstimulation syndrome are described and criteria of predicting the course of this disease and measures to prevent it presented. Statistical data on the presence of this syndrome in various forms during various schemes of superovulation stimulation are offered. Optimal schemes for the treatment of the condition are suggested.     

Remodeling of nuclear architecture during the cell cycle in Drosophila embryos

Little is known about what determines the nuclear matrix or how its reorganization is regulated during mitosis. In this study we report on a monoclonal antibody, mAb2A, which identifies a novel nuclear structure in Drosophila embryos which forms a diffuse meshwork at interphase but which undergoes a striking reorganization into a spindle-like structure during pro- and metaphase. Double labelings with alpha-tubulin and mAb2A antibodies demonstrate that the microtubules of the mitotic apparatus co-localize with this mAb2A labeled structure during metaphase, suggesting it may serve a role in microtubule spindle assembly and/or function during nuclear division. That the mAb2A-labeled nuclear structure is essential for cell division and/or maintenance of nuclear integrity was directly demonstrated by microinjection of mAb2A into early syncytial embryos which resulted in a disintegration of nuclear morphology and perturbation of mitosis.     

Mass transfer during dissolution of a solid into liquid in the iron-carbon system

The rates of dissolution of solid pure iron and a solid iron-carbon alloy into molten iron-carbon alloys were studied “isothermally” as a function of temperature, carbon content and fluid dynamic (stirring) conditions. Experiments were carried out in a resistance-heated tube furnace under an inert atmosphere of argon gas at temperatures ranging from 2165 to 2563°F. A cylindrical specimen of the solid was preheated to the liquid temperature, immersed in the liquid bath contained in a graphite or alumina crucible, and rotated at speeds ranging up to 1800 rpm. The diameter of the specimen after partial solution was determined by direct measurement or calculated from the weight loss of the specimen. The experimental data were fitted with nondimensional correlations of mass transfer for a vertical cylinder, both stationary and rotating. When the specimen was stationary the mass transfer was dominated by natural convection in the liquid bath. When the specimen was rotated, forced convection prevailed and controlled the mass transfer rate. This paper is based in part on a thesis submitted by Y-U. Kim in partial fulfillment of the requirements for the degree Doctor of Philosophy at the University of Michigan.