Cytoplasmic droplets produced by the effect of adenine on Physarum plasmodia: Comparison with caffeine droplets and effect of calcium

Three independently established Drosophila cell lines, Schneider's line 3 (S3), Dübendorfer's line 1 (D1) and MDR3, an adenine salvage deficient clone of the Kc line, all cease to proliferate in the presence of ecdysterone. This is also observed with hybrids between S3 and MDR3 and between D1 and MDR3. It is shown that cells derived directly from wild-type Drosophila embryos can be hybridized with MDR3. Of nine such hybrids all proved to be able to proliferate in the presence of ecdysterone.     

Arabidopsis thaliana as a new model host for plant-parasitic nematodes

We have established culture conditions for successful infection and development of several economically important cyst-forming and root-knot nematodes on Arabidopsis thaliana under monoxenic conditions. Complete life cycles were obtained with the sedentary cyst nematodes Heterodera schachtii, H. trifolii, H. cajani and the root-knot nematodes Meloidogyne incognita and M. arenariaas well as with the migratory nematode Pratylenchus penetrans. In contrast, H. goettingiana and Globodera rostochiensis were unable to develop on Arabidopsis roots. Tissue-culture quality agar and medium conditions optimized for hydroponic root culture were essential for successful infections. Detailed in-vivo observations were made inside Arabidopsis roots during the early infection stages of M. incognita and during complete development of H. schachtii. Seventy-four different ecotypes of Arabidopsis were screened for their susceptibility towards H. schachtii resulting in a range of infection rates. None of the ecotypes tested showed complete resistance in vitro. The use of Arabidopsis as a host for plant-parasitic nematodes will provide a new model system for the molecular genetic analysis of this interaction.     

New approaches to control plant parasitic nematodes

Plant parasitic nematodes are a serious threat for crop production worldwide. This review summarizes our understanding of plant nematode interactions and presents new alternatives for nematode control in the field. Breeding for resistance has been a major goal for many important crop species like soybean, potato, tomato and sugar-beet. As a result numerous nematode-resistance genes have been identified, two of which have been cloned recently, Hs1 ^pro-1 from sugar-beet, giving resistance to the beet cyst nematode Heterodera schachtii, and Mi from tomato, giving resistance to the root-knot nematode Meloidogyne incognita. Also artificial resistance genes, coding for nematotoxic proteins or causing rapid death of feeding cells, have been elucidated. In the future, genetic engineering of nematode resistance will become more and more important for plant breeding. Transformation techniques will allow genes to be quickly introduced into susceptible breeding lines and then combined with each other to produce plant varieties with durable resistance. Received: 26 August 1998 / Received revision: 16 December 1998 / Accepted: 21 December 1998     

Biochemical Characterization of Fungal Phytases (myo-Inositol Hexakisphosphate Phosphohydrolases): Catalytic Properties

Supplementation with phytase is an effective way to increase the availability of phosphorus in seed-based animal feed. The biochemical characteristics of an ideal phytase for this application are still largely unknown. To extend the biochemical characterization of wild-type phytases, the catalytic properties of a series of fungal phytases, as well as Escherichia coli phytase, were determined. The specific activities of the fungal phytases at 37°C ranged from 23 to 196 U · (mg of protein)⁻¹, and the pH optima ranged from 2.5 to 7.0. When excess phytase was used, all of the phytases were able to release five phosphate groups of phytic acid (myo-inositol hexakisphosphate), which left myo-inositol 2-monophosphate as the end product. A combination consisting of a phytase and Aspergillus niger pH 2.5 acid phosphatase was able to liberate all six phosphate groups. When substrate specificity was examined, the A. niger, Aspergillus terreus, and E. coli phytases were rather specific for phytic acid. On the other hand, the Aspergillus fumigatus, Emericella nidulans, and Myceliophthora thermophila phytases exhibited considerable activity with a broad range of phosphate compounds, including phenyl phosphate, p-nitrophenyl phosphate, sugar phosphates, α- and β-glycerophosphates, phosphoenolpyruvate, 3-phosphoglycerate, ADP, and ATP. Both phosphate liberation kinetics and a time course experiment in which high-performance liquid chromatography separation of the degradation intermediates was used showed that all of the myo-inositol phosphates from the hexakisphosphate to the bisphosphate were efficiently cleaved by A. fumigatus phytase. In contrast, phosphate liberation by A. niger or A. terreus phytase decreased with incubation time, and the myo-inositol tris- and bisphosphates accumulated, suggesting that these compounds are worse substrates than phytic acid is. To test whether broad substrate specificity may be advantageous for feed application, phosphate liberation kinetics were studied in vitro by using feed suspensions supplemented with 250 or 500 U of either A. fumigatus phytase or A. niger phytase (Natuphos) per kg of feed. Initially, phosphate liberation was linear and identical for the two phytases, but considerably more phosphate was liberated by the A. fumigatus phytase than by the A. niger phytase at later stages of incubation.     

Glenoid cancellous bone strength and modulus.

The objectives of this study were to determine the strength and modulus of glenoid cancellous bone, including regional variations. The motivations were: to select a suitable bone substitute for standardized testing of glenoid prosthesis loosening, to assist in shoulder prosthesis design and to provide input data for finite element analyses. Ten glenoids from eight cadavers (mean age, 81) were tested by in situ indentation. Mean strength ranged from 6.7 to 17 MPa for the ten glenoids, the overall mean being 10.3 MPa. Mean E moduli ranged from 67 to 171 MPa for the individual glenoids, the overall mean being 99 MPa. These values are likely at the lower end of what would be expected for normal bone since strength and modulus decrease with age and the available specimens were older. These values may be appropriate for prosthesis design, however, since mechanical properties are reduced in rheumatoid arthritic bone. Regional trends were very similar for modulus and strength. The strongest region was postero-superior. The central column, correlating with the keel position in many glenoid components, was weaker than both the anterior and posterior regions but deeper. A large drop in strength and modulus below the subchondral layer emphasizes the importance of maintaining this layer during prosthetic replacement.     

Caffeine Impairs Myocardial Blood Flow Response to Physical Exercise in Patients with Coronary Artery Disease as well as in Age-Matched Controls

Background

Caffeine is one of the most widely consumed pharmacologically active substances. Its acute effect on myocardial blood flow is widely unknown. Our aim was to assess the acute effect of caffeine in a dose corresponding to two cups of coffee on myocardial blood flow (MBF) in coronary artery disease (CAD).

Methodology/Principal Findings

MBF was measured with ¹⁵O-labelled H2O and Positron Emission Tomography (PET) at rest and after supine bicycle exercise in controls (n = 15, mean age 58±13 years) and in CAD patients (n = 15, mean age 61±9 years). In the latter, regional MBF was assessed in segments subtended by stenotic and remote coronary arteries. All measurements were repeated fifty minutes after oral caffeine ingestion (200 mg). Myocardial perfusion reserve (MPR) was calculated as ratio of MBF during bicycle stress divided by MBF at rest. Resting MBF was not affected by caffeine in both groups. Exercise-induced MBF response decreased significantly after caffeine in controls (2.26±0.56 vs. 2.02±0.56, P<0.005), remote (2.40±0.70 vs. 1.78±0.46, P<0.001) and in stenotic segments (1.90±0.41 vs. 1.38±0.30, P<0.001). Caffeine decreased MPR significantly by 14% in controls (P<0.05 vs. baseline). In CAD patients MPR decreased by 18% (P<0.05 vs. baseline) in remote and by 25% in stenotic segments (P<0.01 vs. baseline).

Conclusions

We conclude that caffeine impairs exercise-induced hyperaemic MBF response in patients with CAD to a greater degree than age-matched controls.