L-System model for the growth of arbuscular mycorrhizal fungi, both within and outside of their host roots.

Development of arbuscular mycorrhizal fungal colonization of roots and the surrounding soil is the central process of mycorrhizal symbiosis, important for ecosystem functioning and commercial inoculum applications. To improve mechanistic understanding of this highly spatially and temporarily dynamic process, we developed a three-dimensional model taking into account growth of the roots and hyphae. It is for the first time that infection within the root system is simulated dynamically and in a spatially resolved way. Comparison between data measured in a calibration experiment and simulated results showed a good fit. Our simulations showed that the position of the fungal inoculum affects the sensitivity of hyphal growth parameters. Variation in speed of secondary infection and hyphal lifetime had a different effect on root infection and hyphal length, respectively, depending on whether the inoculum was concentrated or dispersed. For other parameters (branching rate, distance between entry points), the relative effect was the same independent of inoculum placement. The model also indicated that maximum root colonization levels well below 100%, often observed experimentally, may be a result of differential spread of roots and hyphae, besides intrinsic plant control, particularly upon localized placement of inoculum and slow secondary infection.

Production of a periplasmic trehalase in Gluconobacter oxydans and growth on trehalose.

Gluconobacter strains are specialized in the incomplete oxidation of monosaccharides. In contrast, growth and product formation from disaccharides is either very low or impossible. A pathway that allows growth on trehalose was rationally designed to broaden the substrate range of Gluconobacter oxydans. Expression vectors containing different signal sequences and the gene encoding alkaline phosphatase, phoA, from Escherichia coli were constructed. The signal peptide that exhibited the strongest periplasmic PhoA activity was used to generate a G. oxydans strain able to utilize the model disaccharide trehalose as a carbon and energy source by expressing the periplasmic trehalase TreA from E. coli. The strain had a doubling time of 3.7h and reached a final optical density of 1.7 when trehalose was used as a growth substrate. In comparison, the wild-type harboring the empty vector and the strain expressing treA without a signal sequence grew slowly to a final OD of only 0.15. The trehalose concentration in treA expressing cultures decreased continuously during the exponential growth phase indicating that the substrate was hydrolyzed to glucose by TreA. In contrast to the wild-type growing on glucose, the treA expression strain mainly formed acetate and 5-ketogluconate as end products rather than gluconate.

Growth model for arbuscular mycorrhizal fungi.

In order to quantify the contribution of arbuscular mycorrhizal (AM) fungi to plant phosphorus nutrition, the development and extent of the external fungal mycelium and its nutrient uptake capacity are of particular importance. We develop and analyse a model of the growth of AM fungi associated with plant roots, suitable for describing mechanistically the effects of the fungi on solute uptake by plants. The model describes the development and distribution of the fungal mycelium in soil in terms of the creation and death of hyphae, tip-tip and tip-hypha anastomosis, and the nature of the root-fungus interface. It is calibrated and corroborated using published experimental data for hyphal length densities at different distances away from root surfaces. A good agreement between measured and simulated values was found for three fungal species with different morphologies: Scutellospora calospora (Nicol. & Gerd.) Walker & Sanders; Glomus sp.; and Acaulospora laevis Gerdemann & Trappe associated with Trifolium subterraneum L. The model and findings are expected to contribute to the quantification of the role of AM fungi in plant mineral nutrition and the interpretation of different foraging strategies among fungal species.

Minimal early mortality in CABG--simply a question of surgical quality?

BACKGROUND: The increasing number of risk scores and models for the evaluation of the early risk after cardiac surgery reflects the interest in 'calculating' the risk of adverse events. Different time intervals, but also different 'types' of death are generally accepted in the evaluation of early mortality. The aim of this study was to focus on the differences in the calculation of early mortality and to focus on their potentially misleading impact on risk stratification. METHODS: We investigated 7,436 patients who underwent coronary artery bypass grafting from June 30, 1988 through June 30, 2001. A follow-up was performed 180 days after operation (98.7 % complete). RESULTS: According to the definition of 30-day mortality to represent the total time interval between an intervention and the 30th postoperative day, the 30-day mortality was 5.92 % (n = 440 patients). Hospital mortality reflects the number of deaths from the day of intervention through the patient's individual discharge, independent of any fixed time interval. Hospital mortality was 5.86 % (n = 436 patients) in our patient group. 30-day hospital mortality requires the investigation of hospital mortality until the 30th postoperative day; in-hospital and general mortality after the 30th postoperative day remained excluded from the analysis; 30-day hospital mortality was 5.19 % (n = 386 patients). Assuming a maximum hospital stay of 5 days, hospital mortality would decrease to 2.64 % (n = 196 patients). CONCLUSIONS: 30-day mortality, hospital mortality and 30-day hospital mortality are used to determine early outcome. The present data indicate the vulnerability of non-standardized time intervals to discharge policy. However, both hospital mortality and 30-day hospital mortality are predominantly used in current risk scores and models. In view of the comparability and meaning of data, the methodology for the evaluation of early risk should be reconsidered.

Does the completeness of revascularization contribute to an improved early survival in patients up to 70 years of age?

OBJECTIVE: In the era of a renewal of incomplete revascularization approaches, the controversy reappears as to whether the approach for complete revascularization is of prognostic value. The clear advantage of complete revascularization in elderly patients has recently been published. However, for the younger patient group, there is no conclusive information available so far. The aim of our study was to investigate the effect of complete vs. incomplete revascularization in patients up to 70 years of age. PATIENTS AND METHODS: 6531 patients underwent isolated CABG. 5003 of these patients were aged up to 70 years at the time of operation. RESULTS: Incomplete revascularization was performed in 534 (10.7 %) patients. The most common reasons for incomplete revascularization were small vessels and massive calcification. The differences in mortality up to the 180(th) day after CABG are statistically significant. By Kaplan-Meier analysis, the time relationship between incomplete revascularization and death affects predominantly the very early period after CABG. By logistical regression, incomplete revascularization was found to be an independent risk factor for death after CABG. CONCLUSION: Incomplete revascularization affects the early outcome after CABG in patients up to 70 years of age as an independent risk factor for death. In view of recent approaches for primarily incomplete CABG, our results indicate the necessity to reconsider the advantages of complete revascularization.

Does the completeness of revascularization affect early survival after coronary artery bypass grafting in elderly patients?

OBJECTIVE: Usefulness and risks of incomplete versus complete revascularization are still matters of ongoing discussions. Because an increasing number of elderly patients are undergoing coronary artery bypass grafting (CABG), the question arises whether a less extensive surgical approach is more prudent than complete revascularization. METHODS: Of 6531 patients undergoing isolated CABG, 859 were 75 and older at the time of operation. Mean age of the 859 patients was 77+/-2.7 years (median: 76 years); 65% were men. Follow-up enquiry by questionnaire was performed at the 180th postoperative day with a completeness of 95.6%. Assessment of the impact of incomplete revascularization utilized both multivariable analysis and propensity score matching to account for selection factors. RESULTS: Incomplete revascularization was performed in 133 patients (16%). The most common reasons for incomplete revascularization were small vessels (55%) and massive calcification (32%). Mortality until 180 days after CABG was higher (n=32; 24%) after incomplete than after complete revascularization (n=105; 15%; P=0.005). By logistic multivariable regression, incomplete revascularization was identified as an independent risk factor for death (Odds ratio, 1.8; P=0.015). By time-related analysis, incomplete revascularization predominantly affected the early period after CABG (P=0.001). Aortic cross clamping time was only slightly shorter for the group with incomplete (59+/-27 min (median: 55 min) vs. 63+/-26 min (median: 58 min); P=0.1). CONCLUSIONS: Incomplete revascularization increases the early risk of death after CABG in patients aged 75 years and older. The potential compensating benefit of the shorter aortic cross clamping time does not outweigh the advantages of complete revascularization. Thus, in the era of high-volume interventional approaches and minimally invasive techniques, the advantages of complete revascularization need to be considered.

Escherichia coli O157 infections and unpasteurised milk.

We report on two children with Escherichia coli O157 infection, one of whom developed haemolytic uraemic syndrome (HUS). Both had drunk raw cows or goats milk in the week before their illness. Molecular subtyping identified a sorbitol fermenting Escherichia coli O157:H isolate from a dairy cow. This isolate differed from Shiga toxin producing O157:H strains isolated from the 6 year old boy with HUS. This result underlines the need to search for other causes of infection, despite documented consumption of unpasteurised milk. In the second patient, human sorbitol non-fermenting O157:H isolates and animal isolates from goats were indistinguishable. The isolation of indistinguishable sorbitol non-fermenting Escherichia coli O157:H from contact animals supports the association between HUS and consumption of raw goats milk, and re-emphasises the importance of pasteurising milk.

Target, application, and interpretation of scores and alternative methods for risk assessment in cardiac surgery.

BACKGROUND: The more popular the use of different methods for risk adjustment becomes, the more often data are applied without any regard about the primary target and/or about important assumptions. Furthermore, risk adjustment is no longer restricted for quality assurance purposes, but became a "tool" of health policy. Few working groups currently use risk adjustment for the development of new therapeutic concepts. The aim of our study is to clarify possibilities and limitations of popular risk adjustment methods. PATIENTS AND METHODS: 4985 Patients underwent isolated CABG. Statistics was performed by calculating descriptive statistics, Parsonnet, and Higginsscores. Furthermore, the parametric, time-adjusted hazard function by Blackstone was used. RESULTS: Descriptive statistics allows intra-, and interinstitutional comparisons of single items to identify "outlying" results. Risk scores aim to predict preoperatively the risk category of the patient who undergoes cardiac surgery. However, since different scores are based on a score-specific combination of variables, and different definitions of the investigation interval, different results may occur, when different scores are calculated for a single patient. However, the use for example, of scores in patient groups allows description of changing risk structures. Most of the scores derive from univariate analyses and monophasic functions. However, survival curves are predominantly multiphasic and require a consideration of the time-dependency of "risk factors". DISCUSSION: An increasing number of patients with severe comorbidity undergoes cardiac surgery. To evaluate reliably present and futurous therapeutic options, risk adjustment is necessary. Since various tools for risk-adjustment are available, a serious discussion about reliability and application is necessary.

Responses of the soil microbiota to elevated CO2 in an artificial tropical ecosystem.

Plants in artificial tropical ecosystems were grown under ambient (340 microl l(-1)) and elevated (610 microl l(-1)) atmospheric CO2 for 530 d under low-nutrient conditions on a substrate free of organic C. At the end of the experiment a number of soil chemical and microbiological variables were determined. Although we found no changes in total soil organic matter under elevated CO2, we did find that after physical fractionation the amount of organic C in the supernatant (< 0.2 microm) and the amount of water extractable organic C (WEOC) was lower under elevated CO2. The extractable optical density (OD) indicated a higher degree of humification for the elevated than for the ambient CO2 samples (P = 0.032). Microbial biomass C was not significantly altered under high CO2, but total bacterial counts were significantly higher. The microbial biomass C-to-N ratio was also higher at elevated (15.0) than at ambient CO2 (10.0). The number of mycorrhizal spores was lower at high CO2, but ergosterol contents and fungal hyphal lengths were not significantly affected. Changes were found neither in community level physiological profiles (CLPPs) nor in the structural attributes (phospholipid fatty acids, PLFAs) of the microbial community. Overall, the effects on the soil microbiota were small, perhaps as a result of the low nutrient supply and low organic matter content of the soil used in our study. The few significant results showing changes in specific, though relatively minor, organic matter pools may point to possible long-term changes of the more major pools. Furthermore, the data suggest increased competition between plants and microbes for N at high CO2.

Critical illness polyneuropathy: a new iatrogenically induced syndrome after cardiac surgery?

OBJECTIVE: Critical illness polyneuropathy (CIP) is a newly described severe complication after open heart surgery leading to tetraplegia for weeks to months. The purpose of the study was to gather further information on critical illness polyneuropathy developing in patients after cardiac surgery and to evaluate the hypothetical risk factors possibly related to the onset of this neurological disorder. METHODS: From July 1994 to October 1995, 7 out of 1511 patients undergoing open heart surgery developed critical illness polyneuropathy, which was diagnosed on the basis of electromyographic and nerve conduction features. The only common clinical finding was an intensive care unit (ICU) stay beyond seven days, therefore a similar group of 37 patients staying longer than seven days in the intensive care unit during the same period of time was evaluated and retrospectively compared to the 7 patients developing critical illness polyneuropathy. Univariate analysis of several traits was performed to evaluate possible risk factors. RESULTS: 4 Out of 7 patients in the CIP group died, all due to multiple organ failure, in contrast to 3/37 patients in the control group, again due to multiple organ failure. Patients developing CIP were staying significantly longer in the ICU (62+/-3 versus 14+/-8 days, P < 0.01) and had a significantly longer time on ventilator support (50+/-28 versus 7+/-13 days, P < 0.01) The incidence of sepsis was significantly higher in the CIP group than in the control group (85.7 versus 10.8%, P < 0.01). Compared to the control group the proportion of patients receiving corticosteroids (100 versus 10.8%, P < 0.01) and increased dosages of epinephrine and norepinephrine was higher in the CIP group (85.7 versus 35.1%, P < 0.05). Furthermore, the proportion of patients requiring chronic venovenous hemodiafiltration was significantly elevated in the CIP group (85.7 versus 5.4%, P < 0.01). CONCLUSIONS: CIP, despite it's benign nature due to it's spontaneous remission in patients who survive, is a disturbing complication following cardiac surgery which is associated with high mortality, a prolonged stay in the ICU, as well as an extended time on ventilator support. Interventions like chronic hemodiafiltration, the application of corticosteroids and the administration of high doses of catecholamines are more frequent in patients with CIP. Whether this indicates a causal relationship remains to be elucidated.

Scutellospora calospora (Nicol. & Gerd.) Walker & Sanders associated with subterranean clover produces non-infective hyphae during sporulation.

Subterranean clover (Trifolium subterraneum L.) inoculated with either Scutellospora calospora (Nicol. & Gerd.) Walker & Sanders or Glomus sp. (WUM 10(1)) was grown in 3 kg of Lancelin sand surrounding a fine mesh bag (< 38 µm) filled with 400 g of steamed sand. The mesh size was chosen to allow hyphae to penetrate but to exclude plant roots. Every week, from 3 wk after planting the original plants, subterranean clover seeds were sown into the sand inside the mesh bags. The last sowing date was 8 wk from sowing of the original plants. The seedlings, with their roots growing inside the mesh bags, were allowed to grow for 3 wk. At that stage the seedlings and the original plants were harvested and their roots assessed for mycorrhizal colonization. The percentage of root length of the original plants colonized by S. calospora decreased from the first harvest onwards, while colonization by Glomus sp. maintained a similar level at all harvests. Seedlings growing in pots whose original plants were colonized by S. calospora had a her percentage of root length colonized at the first two harvests but levels of Colonization decreased substantially thereafter. Seedlings in the Glomus sp. treatment on the other hand, had similar levels of colonization at all but the last harvest. The length of hyphae g-1 soil in the mesh hues increased throughout the experiment for both fungi. However, the length of hyphae in the S. calospora treatment increased at a faster rate. Spore production by S. calospora commenced 7 wk after sowing the original plants. These results indicate that the hyphae produced by S. colospora after internal colonization had stopped increasing were largely non-infective, and that these hyphae are primarily involved in the production of spores.

Scutellospora calospora (Nicol. & Gerd.) Walker & Sanders associated with subterranean clover: dynamics of colonization, sporulation and soluble carbohydrates.

Two time-course experiments were conducted to investigate the relationship between colonization and sporulation by Scutellospora calospora (Nicol. & Gerd.) Walker & Sanders colonizing roots of subterranean clover (Trifolium subterraneum L.). The percentage of root length colonized by S. calospora increased up to 49 d in expt 1 and 32 d in expt 2 after which it declined. At this stage, there was no further increase in mycorrhizal root length but a sharp increase in non-mycorrhizal root length. In expt 1, the soluble carbohydrate concentration of the roots decreased during the first 49 d, followed by a sudden increase, coinciding with the decline in the percentage of root length colonized. One week before this the shoot P inflow also decreased. In both experiments, sporulation by S. calospora occurred after the decline in colonization. During sporulation there is no further spread in colonization within the root, which may result in increases in soluble carbohydrate concentration within the root. Spore formation may be aided by carbon stores located in the external auxiliary bodies formed by species of the genus Scutellospora.

Cytogenetic studies in a canine mammary tumor.

In a 9-year-old female dog (Basset Artesian Normand) with a mammary adenocarcinoma, cytogenetic evaluation of tumor cells showed a chromosome number of 76 in most metaphases (95%). The following abnormalities were found: symmetric metacentric chromosomes 1 and 6, centric fusion 3/38, a marker X-chromosome (Xmar) and a biarmed small marker chromosome (mar). In the remaining metaphases (5%) there were additional biarmed marker chromosomes present.

[Mechanism of action, scope of the damage and reduction of side effects in intraocular Nd:YAG laser surgery]. / Wirkungsmechanismen, Schadensreichweite und Reduzierung von Nebenwirkungen bei der intraokularen Nd:YAG-Laserchirurgie.

The damage mechanisms of intraocular Nd:YAG laser surgery and their respective damage ranges were investigated in vitro using bovine cornea specimens as a model tissue. The main damage mechanisms are plasma formation and expansion, emission of acoustic transients, and cavitation with jet formation. When a sequence of laser pulses is applied, the interaction of the acoustic transients with gas bubbles remaining from preceding laser exposures is also important. To distinguish the effects caused by the different physical mechanisms, laser pulses were aimed directly onto the corneal endothelium, and parallel to the cornea at various distances. Simultaneously, the cavitation bubble size was determined. The surface morphology and sections of the same lesions were studied by light and electron microscopy. The primary surgical mechanism is tissue evaporation by the laser plasma, whereas the collateral damage from single laser pulses is mainly caused by the cavitation and jet formation. The damage range after a 4-mJ laser pulse is 0.8 mm which is slightly larger than the corresponding cavitation bubble radius. The damage range of the acoustic transients produced by a 4-mJ laser pulse is several millimeters, when they can interact with small gas bubbles attached to the corneal endothelium. The damage range of the acoustic transients alone is smaller than that of cavitation as far as damage detected by light and scanning electron microscopy is concerned. However, on a subcellular level the acoustic transients may possibly cause damage up to a much larger distance. The damage range observed varies with the cube root of the laser pulse energy. A reduction of collateral effects therefore requires the use of small pulse energies. For energies of less than 1 mJ, the pulse duration has to be reduced to ensure plasma production. It is proposed to use low-energy picosecond pulses with moderate repetition rate instead of single nanosecond pulses to reduce collateral damage effects.