Factors Affecting Enhanced Permeation of Amphotericin B Across Cell Membranes and Safety of Formulation

The aim of this study was to determine amphotericin B (AmB) permeation across lipid bilayer membranes mounted on Transwell® and to observe the phagocytosis of the AmB and the AmB-lipid formulations by alveolar macrophage (AM) cell lines using a fluorescence microscope. The lipid bilayer membranes were prepared from phospholipid and ergosterol as well as phospholipid and cholesterol in a ratio (67:33 mol%). AmB-lipid formulations were prepared from AmB incorporated with four lipid derivatives during a lyophilization process. In vitro cytotoxicity studies were carried out on kidney cells by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The levels of nitric oxide production by AMs exposed to these AmB-lipid formulations were determined by the Griess reaction. Phagocytosis of the AmB-lipid formulations was carried out using AM cells. The lipid bilayer membranes and AmB-lipid formulations were successfully prepared. In vitro cytotoxicity results showed less toxicity to kidney cells than pure AmB, and a 1,000-fold less production of nitric oxide by NR8383 cell lines was obtained when compared to lipopolysaccharide. Permeation results were two- to fivefold higher than for pure AmB in the ergosterol containing lipid bilayer and two- to fourfold higher than AmB in the cholesterol containing compositions, both of which were enough to kill the fungi according to their MICs and MFCs. AM phagocytosed the AmB-lipid formulations. We suggest that these products especially the AmB-sodium deoxycholate sulfate are potential candidates for targeting AM cells for the treatment of invasive pulmonary aspergillosis.     

Electric Current-Induced Detachment of Staphylococcus epidermidis Biofilms from Surgical Stainless Steel

Biomaterial-centered infections of orthopedic percutaneous implants are serious complications which can ultimately lead to osteomyelitis, with devastating effects on bone and surrounding tissues, especially since the biofilm mode of growth offers protection against antibiotics and since removal frequently is the only ultimate solution. Recently, it was demonstrated that as a possible pathway to prevent infections of percutaneous stainless steel implants, electric currents of 60 to 100 μA were effective at stimulating the detachment of initially adhering staphylococci from surgical stainless steel. However, initially adhering bacteria are known to adhere more reversibly than bacteria growing in the later stages of biofilm formation. Hence, the aim of this study was to examine whether a growing Staphylococcus epidermidis biofilm can be stimulated to detach from surgical stainless steel by the use of electric currents. In separate experiments, four currents, i.e., 60 and 100 μA of direct current (DC) and 60 and 100 μA of block current (50% duty cycle, 1 Hz), were applied for 360 min to stimulate the detachment of an S. epidermidis biofilm that had grown for 200 min. A 100-μA DC yielded 78% detachment, whereas a 100-μA block current under the same experimental conditions yielded only 31% detachment. The same trend was found for 60 μA, with 37% detachment for a DC and 24% for a block current. Bacteria remaining on the surface after the current application were less viable than they were prior to the current application, as demonstrated by confocal laser scanning microscopy. In conclusion, these results suggest that DCs are preferred for curing infections.     

Factors Affecting Somatic Embryogenesis from Cotyledonary Explants of Safflower

Frequency of safflower (Carthamus tinctorius L.) somatic embryogenesis, number of somatic embryos per responding explant and somatic embryo maturation and germination were affected by genotype, explant age, carbon source, and ethylene. Among 8 cultivars tested, 7 were embryogenic with varying frequencies. The best response was obtained with cv. Girna. Whole cotyledonary explant from 10-d-old plants was best responding compared to 5- or 15-d-old ones. Among different carbon sources, sucrose at 87.6 mM concentration was most suitable for embryo induction, maturation and germination. Of the different ethylene inhibitors, silver nitrate at 50 [micro ]M concentration significantly increased the embryogenic frequency and also the number of embryos per responding explant. Silver nitrate has pronounced effect on embryo maturation but had no effect on germination.     

Factors Affecting Accuracy of Data Abstracted from Medical Records

ObjectiveMedical record abstraction (MRA) is often cited as a significant source of error in research data, yet MRA methodology has rarely been the subject of investigation. Lack of a common framework has hindered application of the extant literature in practice, and, until now, there were no evidence-based guidelines for ensuring data quality in MRA. We aimed to identify the factors affecting the accuracy of data abstracted from medical records and to generate a framework for data quality assurance and control in MRA.MethodsCandidate factors were identified from published reports of MRA. Content validity of the top candidate factors was assessed via a four-round two-group Delphi process with expert abstractors with experience in clinical research, registries, and quality improvement. The resulting coded factors were categorized into a control theory-based framework of MRA. Coverage of the framework was evaluated using the recent published literature.ResultsAnalysis of the identified articles yielded 292 unique factors that affect the accuracy of abstracted data. Delphi processes overall refuted three of the top factors identified from the literature based on importance and five based on reliability (six total factors refuted). Four new factors were identified by the Delphi. The generated framework demonstrated comprehensive coverage. Significant underreporting of MRA methodology in recent studies was discovered.ConclusionThe framework generated from this research provides a guide for planning data quality assurance and control for studies using MRA. The large number and variability of factors indicate that while prospective quality assurance likely increases the accuracy of abstracted data, monitoring the accuracy during the abstraction process is also required. Recent studies reporting research results based on MRA rarely reported data quality assurance or control measures, and even less frequently reported data quality metrics with research results. Given the demonstrated variability, these methods and measures should be reported with research results.     

Factors Affecting Revegetation of Carex lacustris and Carex stricta from Rhizomes

The revegetation of sedge meadows has been problematic because natural recolonization does not occur under many circumstances and because planted propagules often fail to reestablish successfully. In this study, detached rhizomes of Carex lacustris Willd. and Carex stricta Lam. were transplanted in both fall (September) and spring (May) into three experimental wetlands to determine the effects of both planting season and hydrology on survival and establishment. Each experimental wetland had the same mean water depth across 5% slopes, but one had a constant water depth (0.5 m) throughout the growing season, another fell from a mean depth of 0.75 m to 0.25 m, and a third rose from a mean depth of 0.25 m to 0.75 m. Initial rhizome survival, shoot growth, and soil characteristics were recorded over 2 years. Neither planting proved successful (6.9% versus 0.5%) for C. stricta, a tussock-forming sedge. For C. lacustris, a sedge with spreading rhizomes, spring planting had greater rhizome survival (53.2% survival) than fall planting (0.7%). Since both species initiate new shoots in the fall, they are susceptible to transplant failure during this season. The highest survival rates (71–100%) and plant production (736.0 and 494.5 g/m²) for C. lacustris occurred near the water’s edge in both the constant and falling basins. In the rising basin, establishment and growth of this species was high at all water depths (71–96%; 399 g/m²). C. lacustris grew optimally at the same elevations where rhizome survival was greatest, suggesting that shoots are more sensitive to early-season than late-season water levels.     

Factors Affecting Reproducibility of Numerical Classifications

Results obtained with reference organisms which had been included in each of three different investigations were compared. Positions of the reference organisms in diagrams based on highest-link numerical groupings varied with the composition of the microbial samples studied, but significant correlations were found between the results of different investigations. Correlations between the overall groupings obtained in different studies were often better than the correlations between actual similarity values computed between specific pairs of organisms. Similarity values based on different sets of features showed significant correlations, regardless of the kinds of features involved or of the number of features (above a minimum of about 40). Correlations between values based on totally different features were nearly as good as those between values based on similar features which had been determined by different investigators.     

Some Factors Affecting Undergraduate Academic Achievement

A related series of studies, most of which have been published previously, is described. These studies form a coherent whole and demonstrate the development of a theme, namely, the identification of factors in the student and the medical school which, in their interaction, influenced undergraduate academic performance at one medical school. In the population concerned no reliable positive or negative correlation could be demonstrated between cognitive ability and academic performance, when the former was measured by the Wechsler Adult Intelligence Scale and the Medical College Admission Test, and the latter by the current assessment methods of the medical school. Other factors, including socioeconomic and individual personality variables, are at present under investigation as to their effect on academic achievement. It is emphasized that the results of these studies cannot be regarded as valid for all medical schools, but the methods employed can be generalized.     

Factors Affecting Transformation of Pasteurella novicida

The requirements and characteristics of Pasteurella novicida transformations in liquid suspensions were studied. Transformation frequencies of 0.1 to 0.3% were routinely obtained when recipient cells were harvested from 16-hr agar plates and higher than 1% when logarithmic-phase broth-grown cells were used. Calcium ions were essential for transformations. The deoxyribonucleic acid dose response curve, kinetics of transformation, and pH optimum for transformations were similar to those of other bacterial transformation systems. The genetic relatedness of P. novicida and P. tularensis was established by transforming P. novicida mutants with deoxyribonucleic acid extracted from P. tularensis.     

Factors Affecting Transformation of Micrococcus lysodeikticus

Relatively high transfo mation frequencies were obtained with Micrococcus lysodeikticus cells, previou[ill]sly grown in defined broth, with a tris(hydroxymethyl)-aminomethane-glutamate b uffer containing Mg(++), Ca(++), Ba(++), or Sr(++).     

Factors Affecting Egg-ratio in Planktonic Rotifers

Edmondson’s egg ratio (number of amictic eggs per female) is an important life history variable, which has been in wide use to understand and predict patterns of population growth in planktonic rotifers under field conditions. It is also useful as an indicator of the health of rotifers under culture conditions. Generally, an inverse relationship exists between the egg ratio and the density of females in a population. A number of abiotic and biotic factors influence the egg ratio. For example, temperature can cause marked changes in the egg ratio by influencing the frequency of egg production and the hatching times of parthenogenetic eggs. Also, preferential feeding on ovigerous females of rotifers by invertebrate predators such as Asplanchna will lower the egg ratios of the population. The easy detachment of eggs, as may be the case in some members of the Brachionidae especially during enhanced reproduction when food levels are high, may also cause an underestimation of the egg ratio. In this review, we discuss the egg ratio of selected rotifer species in relation to the role of diel changes in egg production, the frequency or the intensity of feeding, the problems of distinguishing between different egg types and the negative effect of stressors such as toxicants and diet quality.     

Factors Affecting Grain Enlargement in Wheat

When most of the grains were removed from wheat ears the remaininggrains developed increased numbers of aleurone and endospermcells, and the increased grain volume was accompanied by anenlarged endosperm cavity. The giant grains usually synthesizedstarch more rapidly than normal grains. Flag leaf photosynthesiswas unaffected and additional sugar was retained in the culmand glumes. Total nitrogen and free amino acid content increased.Gibberellin and auxin concentrations were greater in the glumesof the partly degrained ears, and auxin in the grains was alsogreater than in equivalent grains from intact ears.     

Factors Affecting Transfection in Bacillus stearothermophilus

The conditions for the infection of Bacillus stearothermophilus 4S with TP-1C phage deoxyribonucleic acid (DNA) are described. Cells from log-phase cultures are the most susceptible to phage DNA infection (transfection). A cellular component (competence factor) which enhances transfection is released into the culture medium during the transition period between the log and stationary phase of growth. Transfection is stimulated in the order of decreasing effectiveness, by Fe(3+), Mn(2+), and Mg(2+). The efficiency of transfection is the highest in cells growing at 60.5 C and does not occur in cells growing at 67 C although the cells are growing normally. A cellular component (competence factor) of this organism, which is released into the culture medium, advances by 40 min some step in the uptake of phage DNA.     

Factors Affecting Clonal Diversity and Coexistence

Recent genetic studies of asexually reproducing fishes in thegenus Poeciliopsis (Poeciliidae) revealed abundant variationin the form of multiple sympatric clones. Recurrent hybridizationsbetween sexual species provides the principal source of clonalvariation. The hybrids are spontaneously endowed with a clonalreproductive mechanism that perpetuates a high level of heterozygosity.Migration within and between river systems, and mutations, alsocontribute to clonal diversity in these fish. Coexistence amongdifferent clones and with the sexual ancestors depends in partupon specializations characteristic of individual clones. Clonalreproduction is an efficient mechanism for freezing a portionof the niche-width variation contained in the gene pool of themore broadly adapted, sexual ancestors. Multiclonal populationsachieve significantly higher densities relative to the sexualforms than do monoclonal populations. This relationship is afunction of the clonal variability upon which natural selectioncan act and upon the capacity of a multiclonal population tobetter exploit a heterogeneous environment through niche diversification.In all-female organisms such as Poeciliopsis, which are dependentupon sexual species for insemination, competitive abilitiesprobably are at a premium in the densely populated pools andarroyos of the Sonoran Desert. Competitive abilities are probablyless important for truly parthenogenetic clones which rely oncolonization abilities to escape from their sexual ancestorsand from other clones.