Karyotype of Norway spruce by multicolor FISH.

The chromosomes (2n = 2 x = 24) of Norway spruce are very large since their size reflects the huge amount of genomic DNA (2C = 30 x 10(9) bp). However, the identification of homologous pairs is hampered by their high degree of similarity at the morphological level. Data so far presented in the literature were not sufficient to solve all the ambiguities in chromosome identification. Several genomic Norway spruce DNA clones containing highly repetitive sequences have been identified and characterised in our laboratory. Three of them were selected for fluorescent in situ hybridization (FISH) experiments because of their strong signals and suitability for chromosome identification: PATR140 hybridized at the centromeric site of three chromosome pairs; PAF1 hybridized in six subtelomeric and two centromeric sites; 1PABCD6 co-localized with the subtelomeric sites identified by PAF1. The statistical analysis of microscopic measurements of chromosomes in combination with the FISH signals of these probes allowed the unambiguous construction of Norway spruce karyotype. We also compared the karyotype of Norway spruce with that of other spruce species to infer the number and kind of rearrangements that have occurred during the evolution of these species.

[Daily dialysis: evaluation of the first year of experience at home and in a limited care center]. / La dialisi quotidiana: valutazione del primo anno di esperienza a domicilio ed in un centro ad assistenza limitata.

BACKGROUND: Among self dialysis treatments, daily dialysis is encountering a growing interest. Aim of this study was to evaluate results of the first year of daily dialysis in our Center. METHODS: Since November 1998, twelve patients started daily dialysis. One patient started RRT on daily dialysis; one patient was in training; 8 were on home dialysis, 3 in the limited care center. Selection of patients was performed according to wide acceptance criteria as for age (range 33-61 years), dialysis follow-up (range 1-23 years), comorbidity (=/>1 comorbid factor present in 8). Dialysis schedule consisted of 6 sessions per week (2-3 hours), blood flow 250-320 ml/min, individualized dialysate. Occasional shift to 3-4 times per week were allowed for logistic or working reasons. RESULTS: Results were analyzed taking into account patient satisfaction and main clinical parameters. In 9/12 the choice of treatment resulted from both clinical reasons and patient preferences, while in 3 was due to clinical indications (1/3 dropped out). The main reasons of choice were logistic or research of the best treatment. The most common fears regarded fistula and needle puncturing. Despite the time unconvenience, the rapidly regained well being was the reason for choosing this treatment. Also in this relatively short follow-up the favorable results reported as for weight gain, blood pressure control and metabolic pattern are confirmed. The few side effects were multifactorial (fistula thrombosis after blood pressure normalization, 2 recurrences of atrial fibrillation). CONCLUSIONS: In conclusion, daily dialysis resulted also in our centre as a promising alternative even in difficult patients.

Daily dialysis Kt/V and flexible schedules: is it possible to control efficiency, when and how?

BACKGROUND: Daily hemodialysis is a promising treatment schedule but uniform criteria for defining efficiency are lacking. METHODS: On our daily dialysis (DD) schedule, duration is flexible (2-3 hours, patients are free to add up to 30 min/session), Qb 250-350 mL/min; dialyser 1.6-1.8 m2. Study was performed on 12 pts on DD for > or = 2 months, with > or = 4 Kt/V on subsequent days, tested in the same laboratory. GOAL: To evaluate variability and identify a simple method for weekly calculation, Kt/V was assessed for 133 sessions. RESULTS: On flexible DD, variability of Kt/V-session is high (relative error 4.9%-22%). On flexible schedules, within the time range chosen (2-3 hours) variability of average hourly Kt/V is lower (standard deviation: min (0.014; max (0.052 hour, relative error 4.9%-10%) allowing calculation of weekly Kt/V (averaging 3 sessions: relative error < 6%) suitable for clinical practice. CONCLUSIONS: Flexible schedules, allowing patients to increase treatment time, are an interesting clinical option, but a challenge for Kt/V assessment.

Henoch-Schönlein purpura after 20 years of renal replacement therapy.

While the clinical impact of the impaired immune response, commonly described in chronic dialysis patients, is still a matter of discussion, it is usually considered that immunological diseases tend to become progressively less active after the start of regular renal replacement therapy (RRT). We reported a case of Henoch-Schonlein Purpura in a 51-year-old male, on RRT for 20 years, 8 on dialysis and 12 with renal graft, because of ESRD of unknown origin (chronic glomerulonephritis?). The acute onset of the syndrome, presenting purpuric rash, abdominal discomfort and asymmetric joint pain with edema and local signs of acute inflammation, was followed by several relapses over a 2 years period. This biopsy proven diagnosis offered an explanation for his chronic renal failure; furthermore, we conclude that, possibly because of the usually good correction of uremic immunodepression by efficient dialysis (this patient's Kt/V ranged from 1.1 to 1.3 according to Lowrie's formula), the possibility of immune diseases should be carefully considered even in long long-term RRT patients.

Modelling the "ideal" self care--limited care dialysis center.

Limited care dialysis is an interesting option, which has gained attention in several settings because of the aging of the uremic cohort. The aim of this study was to assess its potential in the Piedmont region in northern Italy, evaluating patients' and care-givers' preferences and testing them in a mathematical model of organisation. The study was conducted in the satellite unit of a university hospital (200-210 dialysis patients), following 35 patients (15 at home, 20 in the center, 10 on daily dialysis). Opinions were collected with a questionnaire and features identified were empirically tested through a simulation model. Most patients (34/35) preferred a small unit, with a stable caring team. Further options were flexibility of dialysis schedule, multiple treatment options, integrated center/home care. These needs could be met by a flexible organization including conventional dialysis (3/week) and daily dialysis (6/week). We employed a simulation model (ARENA software) to calculate the nurses required for each shift and the opening hours and best schedule for the unit. Addition of daily dialysis (2-3 hours) to two conventional 4-5 hour sessions to increased the number of patients followed or "spared" beds, ensuring flexibility. According to patients' best choice (7 dialysis stations), and to the recorded calls, the needs are for two nurses per shift, two shifts per day and six nurses for up to 30 patients in limited care. In conclusion, small centers with flexible schedules can tailor dialysis to patients' needs. A managerial approach is valuable for testing cost/benefit ratios in specific contexts.

Identification of wild and cultivated sunflower for breeding purposes by AFLP markers.

Amplified fragment length polymorphisms (AFLPs) represent one of the most powerful polymerase chain reaction (PCR)-based markers which enables one to discriminate single plants by DNA analysis. To date this technique has only been applied in cultivated sunflower to detect genetic diversity among oilseed inbred lines. In this article we report the use of AFLP markers to investigate the level of diversity within and between populations of Helianthus argophyllus collected in the Maputo area, Mozambique, both for taxonomic and breeding purposes. Three primer combinations gave the best results with 92 polymorphic fragments and were able to discriminate these wild endemic populations from H. annuus and from one of its interspecific hybrids. Most of the variation (71%) observed was within population, and the dendrogram based on shared fragments did not divide the H. argophyllus genotypes into distinct groups resembling different populations. Moreover the hybrid genotypes formed distinguishable subgroups with the cultivated sunflower genotype, confirming the suitability of this technique for taxonomic and phylogenetic studies. From a breeding point of view, although the 12 populations of H. argophyllus represent a new valuable genetic resource, only two of them possessed most of the variation observed, suggesting that they can be the most promising material for crossing with cultivated sunflower.

Education as a clinical tool for self-dialysis.

Therapeutic compliance and patient education are presently considered crucial parts of end-stage renal disease (ESRD) therapy. In the center where Italian home and self-care dialysis treatment started, an education program was designed as multi-step pathway--following patients from chronic renal failure to dialysis and eventual graft--employing lessons, booklets, and books. Each step was validated in various subsets of patients. Lessons involved two hours of informal discussion on the main aspects of ESRD and renal replacement therapy (RRT); booklets were created from tape recordings of the lessons. Patient participation was good, with 28 of 33 patients on charge in the center for 6 months or more taking part in more than one lesson in 1999. In 16 of 16 patients who answered a questionnaire after two lessons, expressed opinion was "good" to "fair." All asked for further material. With regard to books, 500 copies of the book What does dialysis mean? were given out in the region; this book was validated in 22 patients on peritoneal dialysis (PD) and 18 on hemodialysis (HD). It helped patients to accept dialysis in 65% of cases and to comprehend it in 90%. Four thousand copies of the book Stories, containing 18 interviews on transplantation, were printed, and this book was validated in 21 patients on self-care and 35 on hospital dialysis (potential candidates for graft). Of 56 patients, 53 asked for further material; 19 changed their initial opinion (10 choose transplantation, despite initial skepticism; 9 put off transplantation, despite initial acceptance). On a local scale, the program led 12 of 18 new patients, who followed at least part of the program, to choose self-dialysis (PD, home, and self-care dialysis).

Stent implantation in a left anterior descending coronary artery through an internal mammary artery graft.

We describe a case of balloon angioplasty of a totally occluded left anterior descending coronary artery through a tortuous left internal mammary artery graft. Because of early recurrence of the lesion, we implanted a short Palmaz-Schatz stent with good angiographic and clinical results.

[Anatomo-functional correlations of changes in electrocardiographic repolarization in aortic and mitral valve insufficiency. Comparison of indices based on QRS voltages]. / Corrispettivo anatomo-funzionale delle alterazioni della ripolarizzazione all'elettrocardiogramma nelle insufficienze valvolari aortica e mitralica. Confronto con gli indici basati sul voltaggio del QRS.

Electrocardiographic repolarization changes and voltage criteria for left ventricular hypertrophy were examined, in relation to hemodynamic, echocardiographic and angiographic data. This was done to evaluate their association with abnormalities in cardiac function and structure in 53 patients with chronic aortic regurgitation and 36 patients with chronic mitral regurgitation. No patient showed evidence of coronary artery disease. Of the patients with aortic regurgitation, the 27 patients with an abnormal repolarization pattern at ECG had worse NYHA functional class when compared to the 24 patients with normal repolarization (2.4 +/- 1 vs 1.6 +/- 0.9; p less than .01). They also had greater left ventricular dimensions (end-diastolic volume: 162 +/- 57 ml/m2 vs 109 +/- 15 ml/m2, p less than .01; end-systolic volume: 85 +/- 46 ml/m2 vs 44 +/- 31 ml/m2, p less than .01), lower left ventricular ejection fraction (.50 +/- .12 vs .63 +/- .14; p less than .01), greater left ventricular mass (170 +/- 56 gr/m2 vs 119 +/- 29 gr/m2; p less than .01) and higher end-diastolic left ventricular pressure (21 +/- 11 mmHg vs 11 +/- 8 mmHg; p less than .01). QRS voltage was less closely related to cardiac function and structure and thus, did not modify the conclusions based on repolarization findings alone. Furthermore, repolarization patterns identified patient subgroups with high or low prevalences of previously described predictors of poor surgical outcome. The presence or absence of the "strain" pattern was not related to differences in cardiac structure and function, in patients with mitral regurgitation.(ABSTRACT TRUNCATED AT 250 WORDS)

[Surgical results in mitral insufficiency: prognostic value of the indices of left ventricular function]. / Risultato operatorio nell'insufficienza mitralica: valore prognostico degli indici di funzione ventricolare sinistra.

Left ventricular function is a major prognostic factor in patients with mitral regurgitation, but the ability of echocardiographic and hemodynamic parameters to predict the surgical result is controversial. We investigated the prognostic role of various pre-operative indices of left ventricular function in 23 consecutive patients who underwent successful surgical correction of chronic mitral regurgitation. At a mean follow-up of 20 +/- 16 months, patients underwent echocardiography and radio-nuclide angiography and were grouped according to the post-operative left ventricular ejection fraction. Group A was made up of 16 patients with a left ventricular ejection fraction greater than or equal to .45: they showed post-operative reduction of the left ventricular end-diastolic diameter (from 36.3 +/- 3.2 to 30.5 +/- 4.5 mm/m2; p less than .001) and of the radius/thickness ratio (from 3.5 +/- 0.6 to 2.9 +/- 0.6; p less than .01). In 7 patients (group B), post-operative left ventricular ejection fraction was less than .45 and no significant changes in the left ventricular end-diastolic diameter (from 41.5 +/- 2.7 to 36.9 +/- 6.1 mm/m2; NS) or the radius/thickness ratio (from 3.9 +/- 0.9 to 3.4 +/- 1.0; NS) were observed. During the follow-up all group A patients remained asymptomatic or minimally symptomatic, whereas 2 group B patients died of refractory left ventricular failure. Pre-operative left ventricular volumes and diameters, both at end-diastole and end-systole, were significantly greater in group B patients.(ABSTRACT TRUNCATED AT 250 WORDS)

[Evaluation of the seriousness of aortic valve stenosis: calculation of the valvular area or peak? Reliability of Hakki's formula]. / Valutazione della gravità della stenosi valvolare aortica: calcolo dell'area o gradiente al picco? Affidabilità della formula di Hakki.

The severity of aortic valve stenosis should be assessed by means of the calculation of the valvular area; on the other hand, the routine use of the Gorlin's formula for the aortic area is laborious and time consuming. Recently Hakki proposed a simplified formula (area = cardiac output/square root gradient) for the calculation of valvular areas. This method does not require the assessment of the systolic ejection time or the transvalvular flow; furthermore, the peak systolic gradient instead of the mean gradient may be entered into the formula. We have evaluated the reliability of this formula on 83 patients with aortic valve stenosis either pure or with absent to mild aortic incompetence (angiographically first degree maximum). Twenty-eight patients had isolated aortic stenosis, 55 had associated mitral stenosis and/or mitral or tricuspid regurgitation. Our results show a good correlation between the values of valvular areas obtained by Hakki's formula and those obtained by Gorlin's formula (r = 0.90 in the first group and r = 0.91 in the second group of patients). On the contrary we observed a poor relationship between the peak systolic gradient and the valvular area, with a considerable scatter of the data, especially for low values of peak systolic gradient. We therefore conclude that the assessment of the aortic valve stenosis must be based on the estimation of the valvular area; in our hands the Hakki's formula has proven to be easy and sufficiently reliable for routine diagnostic studies.

[Acute dissection of the thoracic aorta. Proposal of a new surgical technic]. / La dissezione acuta dell'aorta toracica. Proposta di una nuova tecnica chirurgica.

Acute aortic dissection is a challenging surgical disease. Replacement of the supracoronary aorta alone can be followed by recurrent aneurysm formation at the level of the residual aortic root. The Bentall procedure prevents this late complication but intraoperative haemorrhage may be severe and valve replacement is always mandatory. A new surgical technique is presented which has been adopted in seven consecutive patients with no deaths. With this procedure, strengthening of the aortic root is obtained by inserting three Dacron Double Velour patches "between" the internal and external aortic lamina, one for each sinus of Valsalva. The patches override the coronary ostia which are left wide patent, and are anchored directly to the aortic anulus by single mattress sutures which reduce the size of the anulus. Suspension of the valve leaflets to the patches overriding each other at the commissures together with anuloplasty reestablish valve continency. The two aortic stumps are secured with running sutures over the free edge and a tubular Dacron graft is then anastomosed to them. The procedure reinforces the aortic root, will prevent recurrent aneurysm formation and avoids at the same time valve replacement, when unnecessary, and coronary arteries reimplantation.

[Corrected transposition of the great vessels in the adult. Presentation of a case simulating ischemic cardiopathy]. / La trasposizione corretta dei grossi vasi nell'età adulta. Presentazione di un caso simulante la cardiopatia ischemica.

We report the clinical and laboratory findings in a 58 years old woman with corrected transposition of the great arteries (CTGA), who that presented typical angina pectoris. The diagnosis of ischemic heart disease was supported by the history of a previous myocardial infarction. Other findings were a systolic murmur of mild mitral regurgitation, left bundle branch block and enlarged left ventricle on the chest X-ray. Cardiac catheterization showed a corrected transposition of the great arteries (L-malposition with situs solitus); left and right coronary arteries were free of luminal stenosis. We suggest therefore that anginal chest pain may be due to myocardial ischemia induced by discrepancy between myocardial oxygen consumption and coronary blood flow. This complication may occur in patients with corrected transposition of great arteries surviving in adulthood.

[Development of a method for the quantitative evaluation of the regional kinetics of the left ventricle]. / Elaborazione di un metodo per la valutazione quantitativa della cenetica regionale del ventricolo sinistro.

Segmental wall motion abnormalities of the left ventricle frequently occur in ischemic heart disease. An objective, quantitative method is required to ensure the reproducibility of the assessment of left ventricular regional function, especially in evaluating the changes induced by diagnostic and therapeutic interventions. In 32 normal subjects we obtained 30 degrees right anterior oblique left ventriculograms and developed a method based on the following observations. The method should reflect the symmetric uniform motion of the left ventricular silhouette. Only actual contractile motion should be taken into account; therefore, rotatory and translational motion should be compensated for. Passive systolic movement of aortic and mitral valves accounts for the contraction of neighbouring myocardial segments. Left ventricular wall excursion is most often measured by a coordinate system: since the cavity of the left ventricle becomes relatively longer during systole, left ventricular walls contract neither toward a single central point nor toward the long axis; therefore the appropriate origin of the coordinate system will be a segment. Furthermore, as more elongated left ventricular end-diastolic silhouettes appear to show a greater extent of systolic lengthening (we show evidence of this), the length of the segment must be related to the end-diastolic shape. The basic steps of the method are: 1) identification of a symmetry line, from the aortic mid-point to the apex, by connecting the mid-point of 19 diameters perpendicular to the long axis; 2) roto-translation of the end-systolic silhouette so that the end-systolic apex and aortic mid-point lie on the symmetry line.(ABSTRACT TRUNCATED AT 250 WORDS)