The absence of phloem loading in willow leaves

Willow (Salix babylonica L.) is representative of a large group of plants that have extensive plasmodesmatal connections between minor vein phloem and adjoining cells. Because plasmodesmata provide a diffusion pathway for small molecules, it is unclear how sucrose could be loaded from the mesophyll into the phloem against a concentration gradient. In the studies reported here, the minor vein phloem of willow leaves plasmolyzed in approximately the same concentration of osmoticum as the mesophyll. Sucrose concentrations in mesophyll cells were greater than those reported in the literature for aphid stylet exudate from willow stems. Calculated turgor pressures in the mesophyll and minor vein phloem were greater than turgor reported in the literature for sieve elements in the stems of willow. Images of minor veins were not obtained in autoradiographs when attached leaves, or leaf pieces, were provided with ¹⁴CO₂ or [¹⁴C]sucrose. Therefore, no evidence could be found for accumulation of sucrose against a concentration gradient in the minor vein phloem of willow. In these leaves, the mesophyll apparently acts as the “source” for long distance transport of sugar. The mechanism of translocation in willow, and the evolution of phloem loading, are discussed.     

Downregulating the sucrose transporter VpSUT1 in Verbascum phoeniceum does not inhibit phloem loading

Sucrose is loaded into the phloem in the minor veins of leaves before export. Two active, species-specific loading mechanisms have been proposed. One involves transporter-mediated sucrose transfer from the apoplast into the sieve element-companion cell complex, so-called apoplastic loading. In the putative second mechanism, sucrose follows an entirely symplastic pathway, and the solute concentration is elevated by the synthesis of raffinose and stachyose in the phloem, not by transporter activity. Several sucrose-transporting plants have been shown to be apoplastic loaders by downregulating sucrose transporter 1 (SUT1), leading to accumulation of sugars and leaf chlorosis. In this study we compared the effect of downregulating SUT1 in Nicotiana tabacum, a sucrose transporter, and Verbascum phoeniceum, a species that transports raffinose and stachyose. To test the effectiveness of RNAi downregulation, we measured SUT1 mRNA levels and sucrose-H⁺ symport in leaf discs. Mild NtSUT1 downregulation in N. tabacum resulted in the pronounced phenotype associated with loading inhibition. In contrast, no such phenotype developed when VpSUT1 was downregulated in V. phoeniceum, leaving minimal sucrose transport activity. Only those plants with the most severe VpSUT1 downregulation accumulated more carbohydrate than usual and these plants were normal by other criteria: growth rate, photosynthesis, and ability to clear starch during the night. The results provide direct evidence that the mechanism of phloem loading in V. phoeniceum does not require active sucrose uptake from the apoplast and strongly supports the conclusion that the loading pathway is symplastic in this species.     

Darwinian adaptation of proteorhodopsin to different light intensities in the marine environment

Proteorhodopsin, a retinal-binding protein, represents a potentially significant source of light-driven energy production in the world's oceans. The distribution of photochemically divergent proteorhodopsins is stratified according to depth. Here, we present evidence that such photochemical diversity was tuned by Darwinian selection. By using a Bayesian method, we identified sites targeted by Darwinian selection and mapped them to three-dimensional models of proteorhodopsins. We suggest that spectral fine-tuning results from the combined effect of amino acids that directly interact with retinal and those that influence the confirmation of the retinal-binding pocket.     

Evidence for direct carotenoid involvement in the regulation of photosynthetic light harvesting

Nonphotochemical quenching (NPQ) refers to a process that regulates photosynthetic light harvesting in plants as a response to changes in incident light intensity. By dissipating excess excitation energy of chlorophyll molecules as heat, NPQ balances the input and utilization of light energy in photosynthesis and protects the plant against photooxidative damage. To understand the physical mechanism of NPQ, we have performed femtosecond transient absorption experiments on intact thylakoid membranes isolated from spinach and transgenic Arabidopsis thaliana plants. These plants have well defined quenching capabilities and distinct contents of xanthophyll (Xan) cycle carotenoids. The kinetics probed in the spectral region of the S(1) --> S(n) transition of Xans (530-580 nm) were found to be significantly different under the quenched and unquenched conditions, corresponding to maximum and no NPQ, respectively. The lifetime and the spectral characteristics indicate that the kinetic difference originated from the involvement of the S(1) state of a specific Xan, zeaxanthin, in the quenched case.     

Detection of circular polarization in light scattered from photosynthetic microbes

The identification of a universal biosignature that could be sensed remotely is critical to the prospects for success in the search for life elsewhere in the universe. A candidate universal biosignature is homochirality, which is likely to be a generic property of all biochemical life. Because of the optical activity of chiral molecules, it has been hypothesized that this unique characteristic may provide a suitable remote sensing probe using circular polarization spectroscopy. Here, we report the detection of circular polarization in light scattered by photosynthetic microbes. We show that the circular polarization appears to arise from circular dichroism of the strong electronic transitions of photosynthetic absorption bands. We conclude that circular polarization spectroscopy could provide a powerful remote sensing technique for generic life searches.     

Atherosclerotic lesion of the coronary arteries in ischemic heart disease patients with differing levels of insulin, cortisol and somatotropic hormone and the type of reactions of these hormones to physical loading

Plasma insulin, cortisol and somatotropic hormone (STH) levels of coronary patients are related to the extent of atherosclerotic lesion of coronary arteries. The relationship between coronary atherosclerosis and hormonal activity becomes particularly evident through the pattern of hormonal response (a rise or a fall) to rationed exercise. An elevated insulin baseline and reduced postexercise levels are typical for coronary patients with only slightly narrowed (25% or less) coronary arteries. A tendency to higher baseline and postexercise plasma cortisol levels was noted in more severe coronary cases. Increased postexercise plasma STH levels were significantly more frequent in patients with less marked atherosclerotic lesions of coronary arteries.     

Analysis of the differing mechanisms of action of the vagus nerves on the work of the heart

Mechanisms of cardiac acceleration in response to vagal stimulation under some experimental conditions were investigated. The stimulation of one (left or right) or both vagal nerves in pigeons was not shown to produce the acceleration phenomenon either in the presence of atropine effects and stimuli of varying intensity, or with weak and sparse stimulation of intact pigeons.     

Reactive oxygen species in the vasculature: molecular and cellular mechanisms

Accumulating evidence indicates that reactive oxygen species (ROS) play major roles in the initiation and progression of cardiovascular dysfunction associated with diseases such as hyperlipidemia, diabetes mellitus, hypertension, ischemic heart disease, and chronic heart failure. ROS produced by migrating inflammatory cells as well as vascular cells (endothelial cells, vascular smooth muscle cells, and adventitial fibroblasts) have distinct functional effects on each cell type. These include cell growth, apoptosis, migration, inflammatory gene expression, and matrix regulation. ROS, by regulating vascular cell function, can play a central role in normal vascular physiology, and can contribute substantially to the development of vascular disease.     

Sensitivity and exposure to indoor allergens in adults with differing asthma severity.

In asthma, it is uncertain whether there is an association between degrees of exposure to domestic allergens and asthma severity. The pattern of sensitivity and exposure to common indoor allergens was examined in subjects with differing asthma severity. Sensitivity to house dust mite, dog and cat allergen and exposure to Der p 1, Can f 1 and Fel d 1 were assessed by skin prick tests and settled dust analysis in 28 subjects with severe asthma and 28 age- and sex-matched subjects with mild asthma (two declined skin prick test). All severe asthmatic subjects had at least one positive skin test and 20 of the 28 subjects were positive to all three allergens. Fourteen of the 26 subjects with mild asthma who took skin prick tests were positive to at least one, and one of these subjects was positive to the three allergens tested. Except for bedroom Fel d 1, the proportion of severe asthmatics both sensitized and exposed to each allergen at each site was significantly greater than the proportion sensitized and exposed in the mild asthma group. The geometric mean allergen concentrations, with the exception of bedroom Fel d 1, were greater in sensitized severe asthmatics than the sensitized mild asthmatics, which was significant for Der p 1 in bedroom samples and Can f 1 in bedroom and living room samples. These results support an association between the degrees of domestic allergen exposure in sensitized individuals and asthma severity.     

Brain uptake of ketones in rats with differing susceptibility to dietary obesity

The effect of a low- and high-fat diet on the transport of 3-hydroxybutyrate and glucose across the blood brain barrier has been measured in two strains of animals that have a marked difference in the degree of obesity that develops when they eat a high fat diet. The S 5B/Pl rats are resistant to dietary obesity whereas the Osborne-Mendel rats readily develop obesity when eating a high-fat diet. The transport of 3-hydroxybutyrate and glucose across the blood-brain barrier was measured as the ratio of radioactive compound (3-hydroxybutyrate or glucose) to radioactively labeled water by the technique of Oldendorf. The uptake of 3-hydroxybutyrate was significantly higher in the S 5B/Pl rats than in the Osborne-Mendel rats when they were eating either the low-fat diet or the high-fat diet. In addition, there was a significant increase in the transport of 3-hydroxybutyrate in the animals of both strains when eating the high-fat diet as compared to the low-fat diet. However, there was no difference in the transport of glucose between the two strains of rats whether they ate a low-fat or high-fat diet. These data are consistent with the hypothesis that resistance to dietary obesity is associated with increased transport of 3-hydroxybutyrate across the blood-brain barrier.     

Resistance of Candida species to antifungal agents: molecular mechanisms and clinical consequences

Candida albicans and related species pathogenic for man become resistant to antifungal agents, in particular triazole compounds, by expression of efflux pumps that reduce drug accumulation, alteration of the structure or concentration of antifungal target proteins, and alteration of membrane sterol composition. The clinical consequences of antifungal resistance can be seen in treatment failures in patients and in changes in the prevalences of Candida species causing disease. These effects were seen unequivocally in HIV-infected patients with oropharyngeal candida infections, but their incidence has decreased dramatically with the introduction of highly active antiretroviral therapy. The evidence for similar emergence of antifungal-resistant yeast strains and species in other types of candida infections is confounded by non-standardised susceptibility testing methods and definitions of a resistant fungal isolate. Recent large-scale surveys of yeasts isolated from blood cultures, based on standardised methodology and resistance definitions, do not support the view that antifungal resistance in pathogenic yeasts constitutes a significant or growing therapeutic problem.     

Central and peripheral mechanisms of the enhanced hypertension following long-term salt loading in spontaneously hypertensive rats

We evaluated whether or not increased sodium (Na) concentrations of cerebrospinal fluid (CSF) and stimulated activities of brain renin-angiotensin system (RAS) contribute to an enhanced hypertension by salt overload in spontaneously hypertensive rats (SHR). Long-term salt loading (1% NaCl solution as drinking fluid) accelerated the development of hypertension in SHR, but did not alter the blood pressure (BP) in normotensive Wistar-Kyoto rats (WKY). CSF Na concentration was elevated in uninephrectomized (Nx) group as compared to that in control SHR, while in WKY CSF Na was not influenced by the treatment. A fall in BP by intravenous AVP antagonist or hexamethonium was greater in salt-loaded SHR than in controls. This hypotensive response to the combined blockade of AVP and SNS correlated with CSF Na in SHR but not in WKY. Plasma concentration of AVP and epinephrine tended to increase in relation to the degree of salt loading in SHR but not in WKY. Pressor responses to intracerebroventricular (ICV) angiotensin II (AII) and NaCl were greater in SHR than in WKY, although these responses were not influenced by chronic salt load in either SHR or WKY. The enhanced hypertensive action of ICV NaCl in SHR was abolished by pretreatment with ICV AII antagonist. Chronic saline drinking enhanced the depressor effect of ICV captopril in SHR but not in WKY. These observations suggest that salt overload in SHR may cause an elevated CSF Na concentration and an enhanced activity of brain RAS, which may increase activity of SNS and release of AVP, resulting in an enhanced development of hypertension.     

Multiple pathways for ultrafast transduction of light energy in the photosynthetic reaction center of Rhodobacter sphaeroides

A pathway of electron transfer is described that operates in the wild-type reaction center (RC) of the photosynthetic bacterium Rhodobacter sphaeroides. The pathway does not involve the excited state of the special pair dimer of bacteriochlorophylls (P*), but instead is driven by the excited state of the monomeric bacteriochlorophyll (BA*) present in the active branch of pigments along which electron transfer occurs. Pump-probe experiments were performed at 77 K on membrane-bound RCs by using different excitation wavelengths, to investigate the formation of the charge separated state P+HA-. In experiments in which P or BA was selectively excited at 880 nm or 796 nm, respectively, the formation of P+HA- was associated with similar time constants of 1.5 ps and 1. 7 ps. However, the spectral changes associated with the two time constants are very different. Global analysis of the transient spectra shows that a mixture of P+BA- and P* is formed in parallel from BA* on a subpicosecond time scale. In contrast, excitation of the inactive branch monomeric bacteriochlorophyll (BB) and the high exciton component of P (P+) resulted in electron transfer only after relaxation to P*. The multiple pathways for primary electron transfer in the bacterial RC are discussed with regard to the mechanism of charge separation in the RC of photosystem II from higher plants.     

Complete amino acid sequence of light chain variable regions derived from five monoclonal anti-p-azophenylarsonate antibodies differing with respect to a crossreactive idiotype.

The induced antibody response to the hapten p-azophenylarsonate in the A/J mouse has provided a model system for the detailed examination of a heritable crossreactive idiotype and its fine structural and serologic analysis. While earlier studies used to apparent homogeneity in the serum response for structural studies, a more complete understanding of the arsonate idiotypic system became possible with the development of monoclonal antibodies differing with respect to these determinants. Five monoclonal antibodies, four crossreactive idiotype positive and one crossreactive idiotype negative, were selected for complete amino acid sequence analysis. The sequences of the light chain variable regions of these molecules are presented here. The data indicate considerable sequence divergence of the monoclonal light chains from the serum light chains. However, there is a striking degree of homology among the monoclonal light chains regardless of the idiotype character of the parent molecule. Although minor variations are apparent throughout the variable regions, the joining regions are identical among light chains in all of these anti-arsonate antibodies. A particularly notable focus of variation is found at positions 92 and 93 in the third hypervariable region. The possible role of this region in the contribution of the light chain to the arsonate crossreactive idiotype is discussed. These data are consistent with the concept that the anti-arsonate monoclonal light chains originate from the joining of a specific J kappa gene segment to a single germ-line V kappa gene segment. These coding segments are likely further subject to a variety of somatic alterations that generate the modest sequence diversity found among the final protein products.     

Targets and endpoints in ablation therapy for atrial fibrillation in the light of pathophysiological mechanisms

Atrial fibrillation (AF), an important public health problem is a complex and multifactorial arrhythmia. Non-pharmacological treatment for symptomatic patients is of increasing importance. The different catheter ablation techniques in AF treatment developed during recent years, all based on different pathophysiological insights, are discussed. The non-standardized use of different follow-up methods after ablation make interpretation and comparison of results difficult.     

The effect of bicarbonate on photosynthetic oxygen evolution in flashing light in chloroplast fragments

The ability of bicarbonate ion (HCO₃^-) to stimulate photosynthetic oxygen evolution in maize chloroplast fragments exposed to continuous light depends on light intensity. Stimulation by HCO₃^- is less at low intensities. In HCO₃^--depleted chloroplasts exposed to brief saturating light flashes, period 4 oscillations (in O₂ yield per flash) are damped within three cycles. Readdition of HCO₃^- to these preparations restores the oscillatory pattern to higher flash numbers, indicating that HCO₃^- reduces the probability of “misses” in the photosystem II reaction center. The rate of the dark relaxation reaction S_n^′ → S_n+1 (where S refers to the oxidation state of the oxygen-evolving mechanism and n = 0, 1, or 2), after a photoact in the photosystem II reaction center, is retarded in HCO₃^--depleted chloroplasts compared to the rate for this reaction in depleted chloroplasts to which HCO₃^- has been resupplied. However, the final oxygen-evolving reaction after the accumulation of four positive charges appears to be independent of HCO₃^-. Bicarbonate has no effect on the dark deactivation of the higher oxidation states (S₂ and S₃) of the positive charge-accumulating system. We propose two alternate ways in which the kinetic model of oxygen evolution developed by Kok et al. [(1970) Photochem. Photobiol. 11, 457-475] can be extended to include the action of HCO₃^-.