Use of Poultry Manure Combined with Soil Solarization as a Control Method for Meloidogyne incognita in Carnation.

The effectiveness of a combination of soil solarization and poultry manure (raw or pelletized) amendments for the control of root-knot nematode (Meloidogyne incognita) was tested in carnation (Dianthus caryophyllus) crops grown in in-ground beds under plastic-covered greenhouse conditions in southern Spain. Our trials demonstrated that soil solarization alone did not provide sufficient control of root-knot nematode, because the carnation growing season in this region only partly coincides with the most effective period for solarization, resulting in an insufficient duration of treatment during a key period for effectiveness. Chemical fumigation with 1,3-dichloropropene + chloropicrin prior to planting was effective in reducing nematode population densities in soil. Its effects spanned 9 months after planting, resulting in acceptable crop yields. In comparison, the combination of soil solarization and raw or pelletized poultry manure was slightly less effective than chemical fumigation for control of this pathogen but crop yields after 9 months were similar. However, the higher root gall indices observed after 9 months, in comparison with chemically fumigated plots, indicated the need for a reapplication of the organic manure treatment at the start of each successive growing season.

Erythrocyte glycohydrolases in subjects with trisomy 21: could Down's syndrome be a model of accelerated ageing?

We studied some erythrocyte glycohydrolases, erythrocyte membrane fluidity, plasma hydroperoxides and total antioxidant defences in 23 Down syndrome (DS) individuals in comparison with healthy age-matched and elderly controls. With regard to erythrocyte plasma membrane fluidity, plasma hydroperoxides and total plasma oxidative defences, DS subjects resembled the age-matched controls more than the elderly ones. Membrane glycohydrolases in DS, however, presented a pattern partly similar to age-matched controls and partly to elderly controls. Concerning cytosol glycohydrolases, DS subjects had lower levels of hexosaminidase and N-acetyl-beta-D-glucosaminidase, the latter specific for the hydrolysis of GlcNAc residues O-linked to proteins. In general, erythrocyte membrane and cytosol glycohydrolases decreased during erythrocyte ageing in DS subjects and in all controls. The increased levels of the same enzymes in DS plasma might be attributed to an alteration of their release-uptake mechanisms between the two different compartments, on account of the higher plasma hydroperoxide levels. These findings indicate that erythrocyte ageing in DS differs partially from that of age-matched and elderly controls. In any case, the accelerated ageing seen in DS is no fully comparable to physiological ageing.

Erythrocyte membrane alterations during ageing affect beta-D-glucuronidase and neutral sialidase in elderly healthy subjects.

In this study, a comparison between elderly (>70 years) and young subjects reveals that elder people are subject to a higher oxidative stress, which causes an increase in plasma hydroperoxide levels (18%) and a decrease in antioxidant defenses (25%). Moreover, the marked decrease of the erythrocyte membrane fluidity observed in elderly subjects was likely to affect the behavior of some membrane glycohydrolases. In fact, a significant decrease of beta-d-glucuronidase and neutral sialidase (30 and 50%, respectively) was detected. Activity differences were also observed when erythrocytes were further distinguished according to their biological age. Striking differences between young and elderly subjects were observed for beta-d-glucuronidase and neutral sialidase in young and senescent erythrocytes, respectively. Overall beta-d-glucuronidase decreases with the subjects' age, while neutral sialidase levels are higher in the elderly. This is presumably due to the localization of these enzymes in distinct plasma membrane micro-domains, which are differently peroxidized. A possible role of these enzymes in signaling praecox membrane alterations has also been evidenced.

First Report of Phytophthora cactorum Causing Fruit Rot on Avocado in Spain.

The area of avocado (Persea americana Mill.) orchards in southern Spain has increased recently and is currently at 8,063 ha. Avocado production in this part of Spain was 72,581 t during 2003. During February 2004, apical necrosis was observed on avocado fruits (cv. Hass) in one orchard in Vélez-Málaga, Málaga Province, southern Spain. Dark brown lesions and necrotic flecking of the flesh also were observed on fruits. Isolations from the skin of the fruit previously washed with tap water and disinfested with 20% sodium hypochlorite on potato dextrose agar (PDA) consistently resulted in mycelial colonies. Sporangia produced on V8 juice by successive washing of mycelia with saline solution (1) measured 31 to 37.2 (33.3) × 21.7 to 28.8 (24.2) µm in size. The pathogen was identified as Phytophthora cactorum on the basis of morphological structures (mycelia, sporangia, chlamydospores, and oospores) formed when grown on V8 juice and PDA (2). To confirm pathogenicity, a mycelial suspension was obtained by blending mycelia grown for 1 week on PDA in 200 ml of sterile water. Three healthy avocado fruits were inoculated with the suspension by injection; three other fruits were inoculated by placing a drop of suspension on the unbroken skin of the fruit. The same number of fruit was inoculated as controls using sterile water instead of mycelial suspension. The inoculated fruits were incubated for 5 days in a moist chamber at 24°C in darkness. Spots appeared on all fruits for both inoculation methods, and the pathogen was isolated and identified as P. cactorum. No symptoms appeared on the control fruits. To our knowledge, this is the first report of P. cactorum causing fruit rot on avocado in Spain. References: (1) D. Chen and G. A. Zentmeyer. Mycologia 62:397, 1970. (2) G. M. Waterhouse and J. M. Waterston. No. 111 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1966.

Cambios físico-químicos del almidón durante la nixtamalización del maíz en variedades con diferente dureza de grano / Physicochemical changes in starch during corn alkaline-cooking in varieties with different kernel hardness

Changes in starch characteristics caused by alkaline-cooking in two corn genotypes (hard and soft) were studied in the present work. Color, pH, pasting properties (Brabender viscoamylograph), water sorption, solubility, swelling power and thermal analysis (Differential scanning calorimetry) were determined in starches previously extracted, from raw and alkaline-cooked hard and soft corns. A reduction in maximum viscosity peak, temperature at the endothermic peak and swelling power at 90 degrees C, and an increase in solubility was observed in starches obtained from both hard and soft corn after alkaline-cooking. Starch from hard corn showed an increment in initial pasting temperature after alkaline-cooking. Alkaline-cooking induced higher modifications in hard than in soft corn starch, probably as a result of its endosperm type and grain hardness, although optimum cooking times were used for each corn genotype.

First Report of Stolbur Phytoplasma in Avocado in Spain.

Since its introduction to southern Spain in the late 1970s, avocado (Persea americana Mill.) has become an alternative irrigation crop to more traditional and less productive dry land crops such as olive, almond, and grapevine. Avocado orchards in this region currently cover an area of 7,500 ha, producing 58,000 tons of fruit in 2000, with 42,000 tons exported annually to the European Union. In summer 1999, symptoms similar to those caused by phytoplasmas were observed in a plot of avocado cv. Hass. Symptoms of leaf roll, leaf veinal chlorosis with the leaves becoming small and abnormally red, and dwarfing were irregularly distributed on affected trees. Host species of phytoplasmas, such as Lavandula officinalis and Thymus officinalis, are found (1) in orchards surrounded by Mediterranean forests. Leaves from eight symptomatic plants taken from affected plots and leaves from two symptomless plants taken from a healthy plot were collected in May, July, and October 2000, and analyzed for phytoplasma infection by polymerase chain reaction (PCR) assays. DNA for PCR was prepared from leaf petioles, midribs, or trunk phloem by phytoplasma-enrichment fraction according to Daire et al. (2). The stolbur phytoplasma was detected in trees by PCR using stolbur-specific nonribosomal primer pair stol 4 f/r (3) or by nested PCR with 16S rDNA primers pairs P1/P7 and fU5/rU3. Phytoplasmas were detected only in samples collected in July. Phytoplasmas were detected by universal primers in all symptomatic samples analyzed in July, whereas stolbur-specific primers gave positive results in only 75% of the symptomatic samples. Restriction fragment length polymorphism (RFLP) analysis with enzymes AluI and Tru9I confirmed the phytoplasma belonged to the stolbur group. To our knowledge, this is the first description of a phytoplasma disease of avocado trees in Spain. Stolbur is the most important vector-borne disease caused by phytoplasma in several crops. In Spain, it has been identified in various crops and weeds such as grapevine, pear, tomato, carrot, pepper, chicory, and strawberry, and appears to be ubiquitous in herbaceous plant host in several families. Rigorous control of stolbur in avocado should be implemented as soon as possible to avoid further disease development and subsequent economic damage to this industry. References: (1) A. Batlle et al. Eur. J. Plant Pathol. 106:811, 2000. (2) X. Daire et al. Ann. Appl. Biol. 121:95, 1992. (3) X. Daire et al. Eur. J. Plant Pathol. 103:507, 1997.

Anatomical variations in the human paranasal sinus region studied by CT.

A precise knowledge of the anatomy of the paranasal sinuses is essential for the clinician. Conventional radiology does not permit a detailed study of the nasal cavity and paranasal sinuses, and has now largely been replaced by computerised tomographic (CT) imaging. This gives an applied anatomical view of the region and the anatomical variants that are very often found. The detection of these variants to prevent potential hazards is essential for the use of current of endoscopic surgery on the sinuses. In the present work, we have studied the anatomical variants observed in the nasal fossae and paranasal sinuses in 110 Spanish subjects, using CT in the coronal plane, complemented by horizontal views. We have concentrated on the variants of the nasal septum, middle nasal concha, ethmoid unciform process and ethmoid bulla, together with others of lesser frequency. The population studied showed great anatomical variability, and a high percentage (67%) presented one or more anatomical variants. Discounting agger nasi air cells and asymmetry of both cavities of the sphenoidal sinus, which were present in all our cases, the variations most often observed were, in order, deviation of the nasal septum, the presence of a concha bullosa, bony spurs of the nasal septum and Onodi air cells.

Soil Solarization in Established Avocado Trees for Control of Dematophora necatrix.

Four field experiments on the control of Dematophora necatrix in avocado orchards affected by white root rot were conducted in the Mediterranean coastal area of southern Spain during 1991 to 1994. In the unshaded locations of solarized plots, the maximal temperatures were 35 to 42°C, depending upon the year and soil depth (15 to 60 cm). Temperature increases attributable to soil solarization ranged between 4 and 8°C in unshaded areas, whereas for shaded areas they were approximately 4°C. Inoculum recovery was decreased in root samples buried at 15 to 30 cm in unshaded locations of both solarized and unsolarized plots after 3 to 5 weeks, whereas 4 to 8 weeks of solarization were required for the elimination of the pathogen buried at depths of 45 to 60 cm. In contrast, inoculum recovery ranged from 30 to 60% for samples in shaded locations of unsolarized plots. D. necatrix was not recovered from roots of infected trees in solarized plots sampled 9 months after solarization, whereas recovery from roots in unsolarized plots was similar to levels before solarization. Soil solarization in established orchards was successful in reducing viability of inoculum buried in soil and eliminated inoculum in infected roots of live trees.

Comparison of exercise and dobutamine stress echocardiography in assessing mitral stenosis.

Dobutamine elicited similar hemodynamic response to exercise in 20 consecutive patients with mitral stenosis, and significantly altered management in 6 of them (30%). Dobutamine stress echocardiography is a safe and feasible alternative to exercise in patients with mitral stenosis of mild-to-moderate severity and ambiguous symptoms.

Usefulness of dobutamine echocardiography in the hemodynamic assessment of mechanical prostheses in the aortic valve position.

Dobutamine stress echocardiography is a simple, safe, and readily available method to evaluate prosthetic valve hemodynamics under various flow conditions. A normal response of aortic valve prostheses to dobutamine infusion is a 100% increment in transprosthetic pressure gradients with an unchanged valve area.

Use of dobutamine stress echocardiography in assessing mechanical aortic prostheses: comparison with exercise echocardiography.

BACKGROUND AND AIMS OF THE STUDY: Relatively high resting flow velocities are occasionally seen in asymptomatic patients with apparently normal mechanical aortic prostheses. Doppler echocardiography in conjunction with bicycle exercise or dobutamine have been used to 'unmask' symptoms or abnormal hemodynamics in these patients. The aim of this study was to compare the Doppler-derived hemodynamic response to exercise and to dobutamine in patients with normally functioning mechanical aortic prostheses. METHODS: Bicycle ergometry (from 200-400 kg x m/min) and dobutamine (from 5-40 micrograms/kg/min) in conjunction with Doppler echocardiography were performed in 25 asymptomatic patients (21 men, four women; mean age 61 years) with mechanical aortic prostheses (range: 19 mm to 27 mm) who had normal resting hemodynamics and normal left ventricular function. Aortic valve area (continuity equation) and maximal instantaneous and mean gradients were estimated at rest and at peak stress. RESULTS: Target heart rate was achieved in 10/25 (40%) patients with exercise, and in 21/25 (84%) with dobutamine (p < 0.005). At peak stress, exercise induced a 25% increase in peak flow velocity, a 55% increase in valve gradients, and no significant change in valve area. In comparison, dobutamine induced a 48% increase in peak flow velocity, a 105% increase in valve gradients, and also no significant change in valve area. The flow velocity and pressure gradients at peak stress were significantly higher with dobutamine than with exercise. CONCLUSIONS: In normally functioning aortic valve prostheses, the target heart rate can be reached more often with dobutamine than with supine bicycle exercise. Despite significant increase in the transvalvular gradients, the valve area remained unchanged. The clinical significance of exercise or dobutamine in symptomatic patients with mechanical prostheses is yet to be proven.

Skeletal maturation of wrist and hand ossification centers in normal Spanish boys and girls: a study using the Greulich-Pyle method.

The skeletal maturation of the upper limb distal segment was studied in 239 Spanish boys and girls between birth and 14 years of age. The atlas of Greulich and Pyle, offering standards of skeletal development of white North American children, was used as reference. The results obtained show firstly that in absolute terms, the ossification process is earlier in girls than in boys. This difference is maximum between 5 and 11 years of age. The boys show a delay of around 3 months with respect to their North American opposites. The girls, in contrast, show a better fit to the corresponding (female) standard of the atlas. In general, the highest agreement between bone age and chronologic age is for the ossification centers of the distal epiphysis of the ulna and radius and for those of the metacarpal bones. At the level of the carpus, individual analysis of the ossification centers shows that the capitate and hamate bones in the girls evolve most in accord with chronologic age. At the other extreme, and in both sexes, the ossification centers of the trapezium and trapezoid bones are statistically furthest from chronologic age.

The clinical utility of transesophageal echocardiography in ischemic papillary muscle rupture.

Ischemic papillary muscle rupture is a potentially lethal complication of myocardial infarction. Acute mitral regurgitation, shock with pulmonary edema, may ensue as its result. Transthoracic echocardiography is the initial noninvasive tool used to confirm the diagnosis. Transesophageal echocardiography has overcome some of the limitations of transthoracic imaging, permitting timely surgical management of these critically ill patients.

The antiproliferative activity of c-myb and c-myc antisense oligonucleotides in smooth muscle cells is caused by a nonantisense mechanism.

Smooth muscle cell (SMC) proliferation is thought to play a major role in vascular restenosis after angioplasty and is a serious complication of the procedure. Developing antisense (AS) oligonucleotides as therapeutics is attractive because of the potentially high specificity of binding to their targets, and several investigators have reported inhibition of SMC proliferation in vitro and in vivo by using AS strategies. We report here the results of our experiments on vascular SMCs using AS oligonucleotides directed toward c-myb and c-myc. We found that significant inhibition of SMC proliferation occurred with these specific AS sequences but that this inhibition was clearly not via a hybridization-dependent AS mechanism. Rather, inhibition was due to the presence of four contiguous guanosine residues in the oligonucleotide sequence. This was demonstrated in vitro in primary cultures of SMCs and in arteries ex vivo. The ex vivo model developed here provides a rapid and effective system in which to screen potential oligonucleotide drugs for restenosis. We have further explored the sequence requirements of this non-AS effect and determined that phosphorothioate oligonucleotides containing at least two sets of three or four consecutive guanosine residues inhibit SMC proliferation in vitro and ex vivo. These results suggest that previous AS data obtained using these and similar, contiguous guanosine-containing AS sequences be reevaluated and that there may be an additional class of nucleic acid compounds that have potential as antirestenosis therapeutics.

Microsurgical anatomy of the human ophthalmic artery: a mesoscopic study of its origin, course and collateral branches.

In the present paper we have studied the gross (mesoscopic) anatomy of the ophthalmic a. in humans, using magnification by microsurgical systems to obtain data on the origin and course of this artery and its main collateral branches. Comparison of our results with previous reports indicates that, although the anatomical variations of the vascular system are well known, some patterns of frequency may be emphasised. Thus, the ophthalmic a. was usually found as a collateral branch of the internal carotid a., although other origins were also found. The ophthalmic a., once inside the orbit, followed a course above the optic nerve in most cases. All the collateral branches of the ophthalmic a., with the exception of the muscular branches, showed great constancy.

Stage-specific oligonucleotide uptake in murine bone marrow B-cell precursors.

Fluorescein isothiocyanate (FITC)-conjugated phosphodiester and phosphorothioate oligonucleotides were used in four-color flow cytometry with murine bone marrow cells stained with monoclonal antibody specific for the differentiation markers B220, S7 (CD43), and BP-1 to show possible stage-specific oligonucleotide uptake. Relatively low uptake was observed among pre-Pro- and early Pro-B cells. Late Pro-B- and pre-B cells had increased oligonucleotide uptake, whereas B cells had a lower level. Cell membrane binding of oligonucleotides varied during B-cell differentiation in parallel with internalization, which was documented by confocal microscopy. An FITC-conjugated polyanionic dextran sulfate also showed differentiation-related B-cell association, suggesting the presence of cell membrane binding sites specific for polyanions as opposed to a unique feature of the DNA backbone. Interpretation of antisense experiments in murine bone marrow cells will need to account for the heterogeneous oligonucleotide uptake among differentiating B cells.

Atherosclerotic plaque evolution in the descending thoracic aorta in familial hypercholesterolemic patients. A transesophageal echo study.

We explored the concept that transesophageal echocardiography can be used as a tool to detect, characterize, and study plaque morphology in the descending thoracic aorta. The pattern of atherosclerotic plaques in the descending thoracic aorta in familial hypercholesterolemic (FH) patients was evaluated. Additionally, evolution of plaque characteristics as a result of therapy was analyzed. In a randomized prospective protocol, eight FH patients (five men and three women, aged 23 to 65 years [mean +/- SD, 42 +/- 14 years]) receiving standard therapy (n = 3; baseline low-density lipoprotein [LDL] cholesterol, 222 +/- 71 mg/dL, mean +/- SD) or LDL apheresis (n = 5; baseline LDL cholesterol, 262 +/- 51 mg/dL) were studied. Baseline and follow-up (mean, 12 months) transesophageal echocardiographic studies were performed. Measurements obtained were atherosclerotic plaque area (PA), aortic wall area (WA), total arterial area (TAA), and plaque-to-wall area ratio (PWR). LDL cholesterol decreased in both groups. The greatest severity of plaque was detected at 30 to 35 cm from the incisors (approximately 15 to 20 cm from the aortic arch). The smallest plaques were present at the arch and more distal descending aorta. In the control group, TAA, PA, and PWR did not change significantly (P = NS versus baseline). In the LDL-apheresis group, TAA increased (P < .05 versus baseline), PA decreased in three of five patients (P = NS versus baseline), and PWR fell (P < .05 versus baseline).(ABSTRACT TRUNCATED AT 250 WORDS)

Improvement in the diagnosis of bioprosthetic valve dysfunction by transesophageal echocardiography.

Transthoracic two-dimensional echocardiography has been considered the method of choice for the non-invasive assessment of bioprosthetic valves. Recently, transesophageal echocardiography has provided a better imaging window for detecting valve abnormalities not identified otherwise, but pathologic confirmation is essential to establish the role of new diagnostic techniques in clinical practice. Therefore, the purpose of this study was to evaluate the usefulness of transesophageal echocardiography in determining the etiology of bioprosthetic valve dysfunction. Transthoracic and transesophageal two-dimensional and color-flow Doppler echocardiography were performed on 44 consecutive patients with suspected bioprosthetic dysfunction who underwent surgical intervention. At surgery a total of 63 abnormalities in 44 bioprostheses were identified: 31 (49.2%) flail cusps, 14 (22.2%) vegetations, four (6.3%) abscesses, four (6.3%) dehiscences, and 10 (15.9%) stenoses. Transesophageal echocardiography was superior to its transthoracic counterpart for the detection of flail cusps (96.7% vs. 54.8%, p < 0.01), vegetations (85.7% vs. 42.8%, p < 0.01), abscesses (100% vs. 25.0%, p < 0.05), and dehiscences (100% vs. 25.0%, p < 0.05). All patients with flail cusps had severe regurgitation as evaluated by color-flow Doppler. In bioprosthetic mitral stenosis, both methods provided accurate estimates of valve area, while in bioprosthetic aortic stenosis, transthoracic Doppler velocity measurements allowed calculation of valve area in four of six patients. In two patients with high flow velocities, the valve area was not estimated because of an inability to measure the left ventricular outflow tract dimension. Conversely, the transesophageal approach provided measurement of the outflow tract in all six patients, but Doppler velocities were not obtained in any of them.(ABSTRACT TRUNCATED AT 250 WORDS)

Anatomical study of the branches emerging along the intracavernous course of the internal carotid artery in humans.

The introduction in the sixties of magnification tools in surgery has created the need for a precise anatomical knowledge of morphological structures that was previously unnecessary from a clinical perspective. In the present paper, we have centred our attention on the intracavernous portion of the internal carotid artery (arteria carotis interna) in humans for applicative purposes. With the aid of a surgical microscope and the corresponding microdissecting material we have analysed the branches emerging from this arterial segment. Our results are compared with previous data appearing in the literature, the latter sometimes confusing because of the different terminologies used to refer to these vessels. The elements under study varied greatly, but some basic variational parameters were deduced.

Comparison of cellular binding and uptake of antisense phosphodiester, phosphorothioate, and mixed phosphorothioate and methylphosphonate oligonucleotides.

The effects of phosphorothioate (S-oligonucleotide) or terminal phosphorothioate-phosphodiester (S-O-oligonucleotides) or methylphosphonate-phosphodiester (MP-O-oligonucleotides) modifications on mouse spleen cell surface binding, uptake, and degradation were studied using fluorescein (FITC)-conjugated oligonucleotides. S-oligonucleotides had the highest cell binding and uptake, followed by S-O-, O-, and MP-O-oligonucleotides. Competition studies indicated that S-oligonucleotides have an increased affinity for cell membrane oligonucleotide binding sites, because they could completely block O-oligonucleotide binding at a molar ratio of just 0.1. Uptake of all oligonucleotides was higher in B cells than T cells and was increased by stimulation with the B-cell mitogen, lipopolysaccharide. Although our cells had been purified using conventional techniques to eliminate dead cells, there remained about 5% of cells that were dead or dying, as determined by flow cytometry using propidium iodide staining. Of note, oligonucleotide association with dead cells was approximately 50-fold greater than that with living cells. Confocal microscopy confirmed that the oligonucleotides in living cells were intracellular, and indicated little nuclear uptake by 4 h. While extensive degradation of intracellular O-oligonucleotides was apparent by 4 h, there was no detectable degradation of S-, S-O, or MP-O-oligonucleotides.