Development of a sink-source interaction model for the growth of short-rotation coppice willow and in silico exploration of genotype×environment effects.

Identifying key performance traits is essential for elucidating crop growth processes and breeding. In Salix spp., genotypic diversity is being exploited to tailor new varieties to overcome environmental yield constraints. Process-based models can assist these efforts by identifying key parameters of yield formation for different genotype×environment (G×E) combinations. Here, four commercial willow varieties grown in contrasting environments (west and south-east UK) were intensively sampled for growth traits over two 2-year rotations. A sink-source interaction model was developed to parameterize the balance of source (carbon capture/mobilization) and sink formation (morphogenesis, carbon allocation) during growth. Global sensitivity analysis consistently identified day length for the onset of stem elongation as most important factor for yield formation, followed by various 'sink>source' controlling parameters. In coastal climates, the chilling control of budburst ranked higher compared with the more eastern climate. Sensitivity to drought, including canopy size and rooting depth, was potentially growth limiting in the south-east and west of the UK. Potential yields increased from the first to the second rotation, but less so for broad- than for narrow-leaved varieties (20 and 47%, respectively), which had established less well initially (-19%). The establishment was confounded by drought during the first rotation, affecting broad- more than narrow-leaved canopy phenotypes (-29%). The analysis emphasized quantum efficiency at low light intensity as key to assimilation; however, on average, sink parameters were more important than source parameters. The G×E pairings described with this new process model will help to identify parameters of sink-source control for future willow breeding.

Antithrombotic effect of tissue and plasma type angiotensin converting enzyme inhibitors in experimental thrombosis in rats.

This study compared the antithrombotic effect of plasma angiotensin converting enzyme inhibitors (ACE-Is): captopril (CAP), enalapril (ENA) and tissue ACE-Is: perindopril (PER), quinapril (QUIN) in experimental venous and arterial thrombosis. Normotensive Wistar rats were treated p.o. with CAP (75 mg/kg), ENA (20 mg/kg), PER (2 mg/kg) and QUIN (3 mg/kg) for 10 days. The influence of ACE-Is on coagulation and fibrinolytic systems as well as platelet function was evaluated. The hypotensive effect of ACE-Is was equal in all groups. QUIN maintained the final carotid blood flow at the highest value in comparison to PER and plasma ACE-Is. The arterial thrombus weight was reduced in PER and QUIN groups while venous thrombus weight was also reduced after CAP. Tissue and plasma ACE-Is caused the inhibition of platelet adhesion and aggregation. A reduction of fibrin generation, prolongation of prothrombin time (PT), activated partial thromboplastin time (APTT) and shortening of euglobulin clot lysis time (ECLT) were observed after PER and QUIN treatment. In conclusion, given in equipotent hypotensive doses, tissue ACE-Is exerted more pronounced antithrombotic effect than plasma ACE-Is in experimental thrombosis. The differences between tissue and plasma ACE-Is in terms of their more pronounced inhibition of experimental thrombosis may be related to the intensified activation of fibrinolysis and inhibition of coagulation.

Correlation between 99mTc-bicisate and regional CBF measured with iodo-[14C]antipyrine in a primate focal ischemia model.

The need for an agent to quantitatively measure regional cerebral blood flow in humans using single photon emission computed tomography has led to the development of a new 99mTc-labeled agent: bicisate (ethyl cysteinate dimer). We have utilized an acute stroke model in the baboon to examine the ability of this agent to quantitatively measure regional cerebral blood flow in ischemic tissue. One hour after occlusion of either the left anterior cerebral or the middle cerebral artery, 99mTc-bicisate was administered intravenously, followed 20 min later by the measurement of local cerebral blood flow using iodo-[14C]antipyrine. With use of double-label autoradiography, the distribution of 99mTc-bicisate was correlated with the local cerebral blood flow images. A cerebral blood flow parameter was calculated from the 99mTc-bicisate tissue distribution and the arterial blood tracer concentration using an indicator fractionation model. For cerebral blood flows above approximately 40-50 ml 100 g-1 min-1, 99mTc-bicisate underestimates cerebral blood flow by as much as 20%, while for blood flows below approximately 15 ml 100 g-1 min-1, blood flow is overestimated by the 99mTc-bicisate distribution by an average of 3-4 ml 100 g-1 min-1. This apparent hyperfixation at very low blood flows may be related to a higher extraction of this tracer by ischemic tissue.

In vivo fluorometric measurement of changes in cytosolic free calcium from the cat cortex during anoxia.

A new approach to assess the mean changes in intracellular free calcium [Ca2+]i directly from the cortex in situ is described along with the [Ca2+]i changes during nitrogen anoxia. Following incision of the dura and part of the pia-arachnoid membrane, quin2 acetoxymethyl ester, 100 microM in artificial CSF, was superfused for 60 min onto the cat cortex. A small cortical area was irradiated with ultraviolet rays (350/30 nm) and the changes in the fluorescence and reflectance were recorded microfluorometrically at 506 and 366 nm, respectively. The net change in the quin2-Ca2+ fluorescence was calculated after correction for the hemodynamic artifact and subtraction of the basal NADH change. The quin2-Ca2+ fluorescence began to increase significantly (48.0 +/- 13.4 units; p less than 0.05) 20 s prior to the isoelectric electrocorticogram (ECoG) and remained elevated during nitrogen anoxia. It decreased steeply 7.3 +/- 1.7 s prior to the recovery of the ECoG activity after the animal was reoxygenated. Thus, the changes in the intracellular free calcium preceded those of the ECoG during a reversible anoxic insult, suggesting that the increase in the [Ca2+]i might be related to the electrical failure during anoxia.

Cytosolic free calcium, NAD/NADH redox state and hemodynamic changes in the cat cortex during severe hypoglycemia.

Using indo-1, a fluorescent Ca2+ indicator, in vivo fluorometric measurements were made of changes in cytosolic free Ca2+, NAD/NADH redox state, and hemodynamics directly from the cat cortex during and after severe insulin-induced hypoglycemia. Cytosolic free Ca2+ started to increase when the EEG became isoelectric, remained at a significantly high level (p less than 0.05) during the period of isoelectric EEG (IEEG), and recovered to the control level 6 min following an intravenous infusion of glucose. The NAD/NADH redox state oxidized significantly during IEEG and then recovered rapidly to the control level after the glucose infusion. Local cortical blood volume (LCBV) increased gradually during the progression of hypoglycemia, reaching the maximal level (146 +/- 7%) at the end of IEEG, and then started to recover. The mean transit time (MTT) through the cortical microcirculation was shortened during the IEEG (control: 3.84 +/- 0.41 s versus IEEG: 2.73 +/- 0.17 s, p less than 0.05), whereas it was prolonged during the 30-min recovery period (5.68 +/- 0.58 s, p less than 0.05). Local cortical blood flow calculated from the LCBV and MTT showed a twofold increase 5 min into IEEG (201 +/- 27% of control, p less than 0.05), recovered 15 min into the recovery period, and then decreased to 77% of control (p less than 0.05) by 30 min. The data support the hypothesis that hypoglycemic brain damage might be mediated by an elevation of cytosolic free calcium.

Acute improvement in histological outcome by MK-801 following focal cerebral ischemia and reperfusion in the cat independent of blood flow changes.

The present study reports on the acute effects of MK-801 on the histopathological outcome and blood flow changes during focal cerebral ischemia and reperfusion. In addition, acute changes in the EEG and blood pressure are also reported. In 16 halothane-anesthetized cats, the left middle cerebral artery (MCA) was occluded for 2 h followed by 4 h of reperfusion. Thirty minutes after the onset of ischemia, eight animals were treated with 1 mg/kg of MK-801, while eight animals received saline. Blood flow from the peripheral MCA territory was measured with H2 clearance. There was a comparable reduction in blood flow (down to 20% of control) in the ischemic gyri of the two groups followed by a partial recovery after recirculation. There was a similar decrease in the EEG amplitude over the ischemic central MCA territory in the treated and the untreated group. Treatment with MK-801 induced a burst suppression in the EEG and a transient drop (11.4 +/- 6.5 mm Hg) in the mean arterial pressure. The volume of early ischemic damage decreased by one-third in the MK-801-treated group compared to the untreated one, both in the total hemisphere (from 29 +/- 10 to 20 +/- 5%) and in the hemispheric cortex (range 36 +/- 8 to 24 +/- 13%). A major fraction of this improvement was localized to the middle and posterior parietal (mainly perifocal) regions of the MCA territory. These results show that in our model, MK-801 improves histopathological outcome despite the lack of apparent effect on the cortical blood flow, and an adverse effect on the systemic blood pressure. This is the first report that describes data on a reproducible model of reperfusion after temporary occlusion of the MCA in a cat, extending the findings of the Glasgow group, who observed similar neuroprotection in models of permanent MCA occlusion.

The effect of hyperglycemia on intracellular calcium in stroke.

The effect of hyperglycemia on cytosolic free calcium ([Ca2+]i) during temporary focal cerebral ischemia was investigated in cats using a fluorometric technique. The middle cerebral artery (MCA) was occluded for a period of 1 h, after which the clip was removed. In seven animals, plasma glucose was raised to 500-700 mg/dl by infusion of a 50% glucose solution starting 30 min after MCA occlusion, while eight animals were kept normoglycemic during and following occlusion. MCA occlusion induced a significant, but identical, elevation of the [Ca2+]i signal ratio (400/506 nm) in both the normoglycemic group (from 1.40 to 1.97 +/- 0.34, p less than 0.01) and in the hyperglycemic group (from 1.40 to 2.00 +/- 0.53, p less than 0.01) at the end of the occlusion. Between 10 and 30 min after reopening, the [Ca2+]i signal ratio decreased to control levels in the normoglycemic group (1.40 +/- 0.11 and 1.36 +/- 0.08 at 10 and 30 min after reopening, respectively), but remained elevated in the hyperglycemic group (1.69 +/- 0.18 and 1.65 +/- 0.21 at 10 and 30 min after reopening, respectively). There was a statistically significant difference between the two groups (p less than 0.01). These data suggest that hyperglycemia may be harmful to calcium recovery during the early recirculation period following focal cerebral ischemia.

A high degree of genetic diversity is revealed in Isatis spp. (dyer's woad) by amplified fragment length polymorphism (AFLP).

Genetic diversity in 38 genotypes, representing 28 individual genotypes from five landraces of Isatis tinctoria (three German: Tubingen, Potsdam and Erfurt, one Swiss and one English), five genotypes of Isatis indigotica (Chinese woad) and five genotypes of Isatis glauca, were investigated using AFLP analysis. Five primer combinations detected a total of 502 fragments of which 436 (86.9%) were polymorphic. The level of polymorphism recorded within each species was 29.8, 86.9 and 35.8% for I. indigotica, I. tinctoria and I. glauca, respectively. Clearly, genetic diversity within I. tinctoria was greater than that observed in I. indigotica or I. glauca. Cluster analyses of the AFLP data using UPGMA and PCO revealed the complete separation of the genotypes of each species into distinct groups. I. indigotica separated as an entirely independent group, whereas I. glauca formed a separate cluster within the I. tinctoria group. Indeed, I. tinctoria and I. glauca are more closely related to each other than either is to I. indigotica. In addition, the genotypes of each landrace, apart from one from the English group, were clearly discriminated. However, the anomalous genotype did associate with the rest of its group when it was linked with the Erfurt group. These results provide new and useful information about the make-up of the Isatis genome, which has not previously been evaluated. They will be useful in the selection of plant material for variety development and conservation of the gene-pool.

A genetic linkage map of willow ( Salix viminalis) based on AFLP and microsatellite markers.

The genus Salix (willow) contains a number of species of great value as biomass crops. Efforts to breed varieties with improved biomass yields and resistances to pests and diseases are limited by the lack of knowledge on the genetic basis of the traits. We have used AFLP and microsatellite markers to construct a genetic map of willow from a full-sib cross of the diploid species Salix viminalis (2n = 38). In accordance with a double pseudo-testcross approach, separate parental maps were constructed and merged to produce a consensus map comprising 291 AFLP and 39 willow microsatellite markers. Nineteen poplar microsatellites were also tested in willow. Five of these amplified loci, of which two were mapped. Linkage groups of the consensus map that could be identified in the parental maps are presented here and spanned 1,256.5 cM with an average interval between markers of 4.4 cM.

Cytosolic free calcium and NAD/NADH redox state in the cat cortex during in vivo activation of NMDA receptors.

Activation of the N-methyl-D-aspartate (NMDA) receptors and the concomitant Ca2+ entry have been implicated in neuronal injury in a variety of pathological states. The effects of extracellular Mg2+ concentrations and D,L-2-amino-5-phosphonovaleric acid (APV), a competitive NMDA receptor antagonist on the NMDA-induced responses were investigated in vivo. In vivo fluorometric measurements were made of changes in cytosolic free Ca2+ ([Ca2+]i) and NADH fluorescence directly from the cat cortex using indo-1, a fluorescent Ca2+ indicator. Changes in [Ca2+]i were assessed utilizing the ratio of indo-1 emission at two wavelengths (400 and 506 nm) during excitation with ultraviolet light (340 nm). Application of 100 microM NMDA to the cortex produced a significant increase in the [Ca2+]i signal ratio at physiological concentrations of Mg2+ (1.2 mM). This increase was enhanced in the absence of Mg2+ and was completely blocked either at 5 mM Mg2+ or in the presence of 50 microM APV. The NAD/NADH redox state was initially oxidized, which was also blocked by either high Mg2+ or APV. The application of NMDA elicited characteristic electroencephalogram (EEG) changes consisting of a marked reduction in amplitude and regular spikes (17-20 Hz). These EEG changes did not appear in the presence of APV. In addition to NMDA receptor antagonists, the level of extracellular Mg2+ is a potent physiological modulator of the NMDA response.

Calcium channel blockers in systemic hypertension.

Alterations in transmembrane flux of calcium ions may be playing a role in the pathophysiology of systemic hypertension. Calcium channel blockers have been shown to be effective antihypertensive drugs with excellent safety profiles. They are efficacious in the long term treatment of systemic hypertension in all population subgroups, and have special applicability for treating patients with hypertensive urgencies and individuals with concomitant diseases such as angina pectoris and arrhythmias.

Nasopharyngeal carcinoma. A study examining Asian patients treated in the United States.

From 1972 to 1991, 126 Asian patients with nasopharyngeal carcinoma underwent definitive radiation therapy for locoregional disease: 86 men, 40 women. Median age was 50. All patients received external-beam irradiation with cobalt 60 or 4-18 MV x-rays. Local recurrence, regional recurrence, and distant metastases were 22%, 11%, and 23%, respectively. Local recurrence progressively increased with increasing T stage, but doses in those who recurred did not differ from the group as a whole. Regional recurrence was not associated with T or N stage or dose. Patients with N2 disease had the highest distant metastatic rate. The 5- and 10-year overall survival rates were 54% and 38%, respectively. Of age, gender, and histology, only age less than 50 was found to be favorably prognostic. No severe long-term complications were observed, and acute reactions were acceptable. Our survival results are comparable with results found both in Asia and North America. Nonetheless, altered fractionation techniques and/or other radiation modalities should be further explored to improve locoregional control.

A prospective study of the effect of the appropriateness of foot-shoe fit and training shoe type on the incidence of overuse injuries among infantry recruits.

In a prospective study of the effect of the appropriateness of foot-shoe fit and training shoe type on the incidence of overuse injuries among infantry recruits, it was found that three shoe widths for each shoe length size were necessary to adequately accommodate the recruit population's foot anatomy. Recruits compensated for the lack of available shoe widths by choosing larger shoe sizes. However, this compensation did not result in an increase in the incidence of overuse injuries. Recruits who trained in basketball shoes had a lower incidence of overuse injuries of the feet than recruits who trained in infantry boots (p = 0.0001), but the overall incidence of overuse injuries was the same in both groups.