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A single chitin synthase gene is responsible for chitin production in the eggshells of the plant parasitic nematode Meloidogyne artiellia. In this paper we describe a functional analysis of this gene using RNAi as well as further analysis of two similar genes from the free-living nematode Caenorhabditis elegans. The parasitic life-style of M. artiellia required the development of a novel method for delivery of dsRNA to nematode eggs that may be of utility in other experimental systems. C. elegans chitin synthase genes were silenced by feeding nematodes bacteria expressing appropriate chitin synthase dsRNA from a plasmid vector, while M. artiellia egg masses were soaked in dsRNA solution. The results obtained demonstrated that the synthesis of chitin continues to take place in nematode eggs within the egg sac in the parasitic nematode, and that the removal of this activity affects egg development in both C. elegans and M. artiellia. The method described here provides a new way of investigating gene function in plant parasitic nematodes allowing the validity of potential target genes for novel control methods to be assessed. Furthermore, since intact egg cells within the gelatinous matrix of M. artiellia are permeable to dsRNA, eggs of other nematodes may also be similarly permeable to dsRNA and therefore amenable to use with dsRNAi.

Background: Epidemiological estimates of left ventricular mass are based on echocardiographic imaging from the parasternal view, which is often unavailable in subjects with obesity or lung disease. This study was undertaken to assess whether the subcostal view is a valid alternative to estimate left ventricular mass in an unselected older population., Methods: In a cross-sectional study of all the residents in Dicomano, Italy, aged > or =65 years, echocardiography was performed with a systematic attempt to obtain both the parastermal and the subcostal views., Results: The parasternal view was missing in 73/614 participants, 48 of whom were imaged from the subcostal view. In participants imaged from both views, the subcostal view underestimated left ventricular cavity dimension and, consequently, left ventricular mass [79.7 (1.3) vs. 93.3 (1.5) g/m2; p<0.001]. Furthermore, the subcostal view was only 25% sensitive for the diagnosis of hypertrophy. Several multivariate regression models, developed in an equation development subgroup and tested in a validation subgroup, failed to correct the prediction of left ventricular mass based on measures taken from the subcostal view, also after inclusion of demographic, anthropometric, and spirometric covariates., Conclusions: In unselected older persons, the subcostal view does not improve the accuracy of echocardiographic estimation of left ventricular mass, which remains biased in epidemiological studies.

Background: Diffusion is a physical process based on the random movement of water molecules known as Brownian movement. Diffusion-weighted imaging (DWI) is a magnetic resonance (MR) technique that provides information about the biophysical properties of tissues such cell organization and density, microstructure and microcirculation., Materials and Methods: Twenty healthy volunteers and 18 patients with renal tumor were enrolled in our study. The DWI was obtained before contrast media injection with a single-shot SE EPI Inversion Recovery (IR) sequence. The tumor cellularity of each resected lesion was evaluated., Results: The mean apparent diffusion coefficient (ADC) value of renal tumors was significantly lower than the mean ADC value of normal renal parenchyma. In our series, the mean ADC value of renal tumors did not significantly correlate with tumor cellularity, but correlated with histological architecture., Conclusion: These preliminary results indicate the utility of DWI in the acquisition of tissue characterization data of renal masses using a minimal acquisition time (17 sec).

Haploidentical stem cell transplantation has became a clinical reality in the last 10 years as it provides the chance of transplant for about 50% of patients with hematological malignancies who do not have a matched related or unrelated donor. Proper graft preparation for this type of transplant is crucial and this paper analyses our work over the past decade in the search for the optimal graft processing procedure moving from E-rosetting and soybean agglutination, through a combination of negative or positive selection of hematopoietic stem cells to the current method of one-step positive selection. In preparing a graft for haploidentical transplant, three essential requisites must be met. It must contain (1) a megadose (>10 x 10(6) x kg recipient b.w.) of hematopoietic stem cells to overcome the HLA histocompatibility barrier; (2) very few T-lymphocytes (CD3+ cells < 3 x 10(4)/kg recipient b.w.) to prevent severe acute and chronic graft-versus-host disease (GvHD); (3) very few B-lymphocytes to prevent Epstein-Barr virus-related lymphoproliferative disorders. With current graft processing technologies based on positive selection of hematopoietic stem cells, these requirements can be met. A 70-80% hematopoietic stem cell recovery ensures the target megadose is achieved in over 70% of cases with a T-cell depletion of more than 4 logs and a B-cell depletion of over 3 logs. Progress in graft processing has ensured primary, sustained engraftment rates of over 90% and has significantly reduced the incidence of severe acute GvHD and EBV-related lymphoproliferative disorders. Modern time-saving automated graft processing devices ensure reproducibility, reliability, and biological safety, which make widespread application of the haploidentical transplant currently feasible.

Objectives: We sought to compare construct and predictive validity of four sets of heart failure (HF) diagnostic criteria in an epidemiologic setting., Background: The prevalence estimates of HF vary broadly depending on the diagnostic criteria., Methods: Data were collected in a survey of community dwellers who were > or =65 years of age living in Dicomano, Italy. At baseline, HF was diagnosed with the criteria of the Framingham, Boston, and Gothenburg studies and of the European Society of Cardiology (ESC). Left ventricular mass index and ejection fraction, left atrium systolic dimension, lower extremity mobility disability, summary physical performance score, and 6-min walk test were compared between HF and non-HF participants to test for construct validity of each set of criteria. Predictive validity was evaluated with follow-up assessment of cardiovascular mortality, incident disability, and HF-related hospitalizations. Comparisons were adjusted for demographics, comorbidity, and psychoaffective status., Results: Of 553 participants, 11.9%, 10.7%, 20.8%, and 9.0% had HF, according to Framingham, Boston, Gothenburg, and ESC criteria, respectively. In terms of construct validity, Framingham and Boston criteria discriminated HF from non-HF participants better than Gothenburg and ESC criteria across the measures of cardiac function and global performance. The Boston criteria showed a superior predictive validity because they indicated a significantly greater adjusted risk of cardiovascular death (hazard ratio3.9, 95% confidence interval 1.2 to 13.2), incident disability, and hospitalizations in participants with HF., Conclusions: The Boston criteria are preferable to Framingham, Gothenburg, and ESC criteria for the diagnosis of HF in older community dwellers because they have good construct validity and more accurately predict cardiovascular death, incident disability, and hospitalizations.

Background & Aims: The real importance of lactose malabsorption in irritable bowel syndrome (IBS) is still controversial. The aim is to define the relationship between patient perception of milk tolerance/intolerance, lactose malabsorption and abdominal symptoms in IBS., Methods: A hydrogen breath test (HBT) after an oral load of lactose was carried out in 475 consecutive IBS patients, diagnosed according to the Rome criteria. Data were analyzed in 201 age- and sex-matched pairs of IBS patients, classified according to self-reported milk tolerance/intolerance. Hydrogen peak and excretion, predominant presenting symptom and the occurrence of symptoms during the test, were evaluated., Results: The prevalence of positive HBT and the occurrence of symptoms during the test was similar in milk "tolerant" (68.6%, 40.7% respectively) and "intolerant" patients (75.6%, 42.7% respectively), as well as peak (76.4 vs 75.2 ppm) and amount of H2 excreted (57.8 vs. 53.2 ppm/h). The positive predictive value for self-reported milk intolerance was 0.75, and the negative predictive value in regular milk users was 0.31, reflecting the prevalence of lactose malabsorption in the general population more than the awareness of milk tolerance., Conclusions: In IBS patients, self-reported milk intolerance does not help in identifying lactose malabsorbers. The opposite does not rule out the occurrence of symptoms after a lactose load. Lactose is, indeed, responsible for symptoms in some IBS patients, however, these patients can only be identified by the occurrence of symptoms during the test, and not on the individual perception of milk intolerance.

Objectives: To assess whether kyphosis is associated with ventilatory dysfunction in older community dwellers., Design: Cross-sectional study., Setting: The unselected population of Dicomano, Italy aged>or=65 years., Participants: A total of 323 nonheart failure participants underwent clinical evaluation for the presence of kyphosis and spirometry. The severity of kyphosis was estimated from the difference between standing stature and knee-height-derived stature and from the occiput-wall distance., Measurements: Forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), and prevalence of obstructive and restrictive ventilatory pattern., Results: The 130 kyphotic participants (40.2%) had an adjusted 2.5 prevalence odds ratio (POR) for dyspnea (95% confidence interval (CI)=1.1-5.8). FVC% and FEV1% were lower in the presence of kyphosis (P<.01); their deficit was proportional to kyphosis severity. The ventilatory dysfunction was underestimated when reference spirometric parameters were calculated based on standing stature, compared with knee-height derived stature. Of the kyphotic participants, 56.2%, 26.9%, and 16.9% had spirometric normal, obstructive, and restrictive patterns, respectively. Kyphosis was associated with a restrictive (adjusted POR=2.3, 95% CI=1.1-4.8; P=.021) and an obstructive ventilatory pattern (adjusted POR=3.3, 95% CI=1.7-6.5; P<.001)., Conclusion: In unselected older persons, kyphosis is associated with dyspnea and ventilatory dysfunction of a restrictive and an obstructive type. Kyphosis should be included in the differential diagnosis of dyspnea and ventilatory dysfunction in the elderly.

Objectives: To evaluate whether older persons using angiotensin-converting enzyme (ACE) inhibitors have a larger lower extremity muscle mass (LEMM) than users of other antihypertensive drugs., Design: Cross-sectional analysis of data from the Health, Aging and Body Composition (Health ABC) Study., Setting: University of Tennessee, Memphis, and University of Pittsburgh clinics., Participants: A community-based sample of 2,431 well functioning participants of the Health ABC, aged 70 to 79, who were free of heart failure, were selected according to use of antihypertensive medications: ACE inhibitors (n=197), beta-blockers (n=169), thiazides (n=216), calcium-channel blockers (n=340), or none (n=1,509)., Measurements: LEMM, assessed using dual-energy x-ray absorptiometry, compared by index drug in analysis of variance models unadjusted and adjusted for demographics, study site, height, body fat, physical activity, blood pressure, coronary artery disease, diabetes mellitus, and chronic pulmonary disease., Results: LEMM significantly differed across the study groups, being larger in users of ACE inhibitors than in users of other drugs (unadjusted and adjusted models). LEMM was comparable in users of ACE inhibitors and no drug users. A trend toward larger LEMM was also observed in sex- and ethnicity-stratified analyses and in the subgroup of noncoronary hypertensive participants., Conclusion: In older persons, use of ACE inhibitors is associated cross-sectionally with larger LEMM. This finding suggests a possible explanation of the benefits of ACE inhibitors in wasting syndromes. If confirmed in longitudinal studies, this pharmacological action might have important implications for the prevention of physical disability in older patients with hypertension.

Introduction: High-dose chemotherapy conditioning regimens for autologous stem cell transplantation generally give similar results in multiple myeloma. We compared two regimens: melphalan versus melphalan plus busulphan., Methods: In all, 30 untreated patients with stage III low-risk multiple myeloma were studied. After induction with three VAD courses and mobilization with cyclophosphamide 7 g/m(2) and recombinant human granulocyte-colony stimulating factor (rHuG-CSF) (10 microg/kg b.w./die), they received melphalan 200 mg/m(2) (arm A) or busulphan 16 mg/kg plus melphalan 100 mg/m(2) (arm B) for conditioning for transplantation. All patients received maintenance therapy with Interferon 3 MU x 3/week., Results: Time to engraftment after transplantation was similar in both groups. All patients received rHuG-CSF after reinfusion of peripheral stem cells. No differences emerged in transplant-related infective and noninfective complications. There were no transplant-related deaths. A better response was observed in the melphalan plus busulphan regimen (85 versus 75%, P<0.05). The 5-year overall survival with this regimen was 56 versus 49% with melphalan, and the median survival was 126 months versus 108 months for melphalan (P=0.7). The median progression-free survival was 121 months for melphalan plus busulphan versus 97 months for melphalan (P=0.05)., Conclusion: These two conditioning regimens showed similar overall response rate and overall survival, though progression-free survival was better with busulphan plus melphalan.

Purpose: The latest innovation in MR imaging is SENSE (SENSitivity Encoding), a technology providing an important solution for the time necessary for signal encoding. In the SENSitivity Encoding (SENSE) approach, an array of multiple, simultaneously operated receiver coils is used for signal acquisition. In particular, the efficacy of SENSE technology is able to reduce the number of phase encodings by a factor (named R factor or Reduction factor) and to evaluate the same k space with fewer readout lines obtaining higher spatial and temporal resolution., Materials and Methods: An heterogeneous group of 30 patients with a variety of clinically proven diagnoses underwent Cardiac MR with SENSE technology to evaluate the technique's diagnostic efficacy. The sequences used were "Balanced" Ultrafast Gradient Echo (B-FFE) characterised by a hyper-intense blood signal with multislice multiphase and single slice multiphase acquisitions., Results: In addition to reducing scan times, SENSE technology improved spatial and temporal resolution (40 frames/cardiac cycle) providing optimal dynamic evaluation of valve structures and wall kinesis. Furthermore, the use of the SENSE technique with B-FFE sequences (sB-FFE) enabled the qualitative evaluation of abnormal blood movements and transvalvular flows., Conclusions: The considerable time savings allowed by SENSE technology and the clear improvement in image quality constitute a step forward in cardiac MR imaging. The possibility of executing morphologic, dynamic, perfusional and spectroscopic studies in the same MR examination session with short acquisition times and good image quality are becoming more feasible.

ABSTRACT In the Mediterranean Basin, Fusarium oxysporum f. sp. ciceris and the root-knot nematode Meloidogyne artiellia coinfect chickpea. The influence of root infection (after inoculation with 20 nematode eggs and second-stage juveniles per gram of soil) by two M. artiellia populations, from Italy and Syria, on the reaction of chickpea lines and cultivars with partial resistance to Fusarium wilt (CA 252.10.1.OM, CA 255.2.5.0, CPS 1, and PV 61) and with complete resistance to F. oxysporum f. sp. ciceris race 5 (CA 334.20.4, CA 336.14.3.0, ICC 14216 K, and UC 27) was investigated under controlled conditions. In genotypes with partial resistance, infection by M. artiellia significantly increased the severity of Fusarium wilt, irrespective of the fungal inoculum density (3,000 or 30,000 chlamydospores per gram of soil), except in cultivar CPS 1 at the lower fungal inoculum density. In genotypes with complete resistance to Fusarium wilt, infection by M. artiellia overcame the resistance to F. oxysporum f. sp. ciceris race 5 in CA 334.20.4 and CA 336.14.3.0 but not in ICC 14216 K, irrespective of the fungal inoculum density, and overcame the resistance in UC 27 only at the higher inoculum density. Infection by the nematode significantly increased the number of propagules of F. oxysporum f. sp. ciceris race 5 in root tissues of genotypes with complete resistance to Fusarium wilt, compared with roots that were not inoculated with the nematode, irrespective of the fungal inoculum density, except in ICC 14216 K, in which this effect occurred only at the higher inoculum density. Reproduction of an M. artiellia population from Syria in the absence of F. oxysporum f. sp. ciceris race 5 was significantly higher than that of a population from Italy in all tested chick-pea genotypes except ICC 14216 K. However, there was no significant difference between the reproduction rates of the two nematode populations in plants infected with F. oxysporum f. sp. ciceris race 5, irrespective of the fungal inoculum density and the reaction of the genotypes to the fungus.

Background and Aims: It is acknowledged that, in spite of their generally worse health, women live longer than men. However, whether women also enjoy longer disability-free lives is still unclear. Using data from a representative, Italian cohort followed for 6 years, this study aimed at estimating differences between men and women in the age of disability onset and in total survival., Methods: In 1989, 651 persons aged > or = 65 years were interviewed and their medical status was assessed by a geriatrician. In 1995, the time of onset of disability was reconstructed by re-interviewing 392 survivors and collecting proxy information for 201 subjects who had died. No information was available for 58 subjects who refused to be re-interviewed or were lost to follow-up. Data on changes in functional status were also collected by proxy interview for 34 additional persons who had died during the follow-up period, although they had not been originally interviewed at baseline., Results: Of the 235 deaths, 113 were men and 122 were women. On average, the age at death was 3.5 years higher among women than among men. However, the age at onset of disability was similar in the two sexes. In survival analysis in which age was the time variable, women were as likely as men to develop disability, but significantly less likely to die over the follow-up period., Conclusions: Compared with men, women experience longer disability before death. This may be due to sex-related differences in the lifetime prevalence of lethal vs. disabling diseases.

In mammals, methylation of DNA within regulatory sites and histone deacetylase recruitment in transcriptional repressing domains are involved in the loss of the expression of retroviral DNA or repeat arrays transferred in cells for therapeutic purposes. Various investigation results suggest that methylation/deacetylation events are modulated by extracellular and cytoplasmic signal transduction pathways closely involved in regulating cell differentiation. To analyse gene silencing mechanisms and assess if potential pharmacological treatment affects gene silencing kinetics we transduced U937 myelomonocytic cells with a bicistronic retroviral construct carrying the herpes simplex virus thymidine kinase (HSV-TK) and beta-galactosidase (Lac-Z) genes. This vector can be employed in vivo and in vitro to render transduced cell populations susceptible to ganciclovir (GCV). We verified the effect of the histone deacetylase inhibitor Trichostatin A (TSA) alone or combined with 5'-azacytidine (5'aza-C) on transcription downmodulation. Our results indicate that in our in vitro model TSA is able to reactivate transgene expression, more efficiently and with quicker kinetics (12-24h) than 5'aza-C (36-48 h). The effect is dose dependent (between 1 and 50 nM), with no relevant toxicity. Treatment with both drugs is synergistic in gene reactivation in terms of extension and persistence, with low toxicity and no relevant differentiating effects. The cells in which transgene expression has been reactivated undergo progressive silencing, but once weekly drug treatment can maintain high transgene expression levels for more than 90 days with no evidence of selection. The results obtained by treating U937 transduced clones with TSA and/or 5'aza-C together with IL-3, G-CSF or GM-CSF cytokines suggest that transduced U937 differentiation levels do not affect basal expression, but render these cells more responsive to reactivation by TSA or TSA plus 5'aza-C, but not to 5'aza-C alone. In conclusion, the results suggest that in vitro inhibition of histone deacetylase by TSA can interfere with gene silencing mechanisms affecting 5' Moloney murine leukaemia virus long terminal repeat (MoMuLV-LTR) driven transgene expression thus providing the rationale for TSA and/or 5'aza-C administration in animal models for the translation on gene therapy applications.

Background: Our aim was to prospectively evaluate a semi-quantitative pulmonary perfusion MR technique using a breath-hold 2D dynamic sequence in patients with severe pulmonary emphysema., Materials and Methods: Thirty patients with severe emphysema were studied with pulmonary perfusion MRI. Results were compared with those obtained through lung scintigraphy, considered as the gold standard technique. We used Fast Field Echo (FFE) pulse sequences in the coronal and sagittal planes, with paramagnetic contrast agent administration. Ten healthy volunteers were studied as the control group. Three thoracic radiologists independently reviewed all the subjects, evaluating the site and entity of perfusional defects. Peak intensity and apparent mean transit time were calculated., Results: MRI showed high sensitivity (86.7%) and good specifity (80.0%) in detecting perfusional defects. We observed lower peak signal intensities in emphysematous regions., Conclusion: Lung perfusion MR is a potential alternative to Nuclear Medicine in the evaluation of severe pulmonary emphysema.

Purpose: In previous studies, higher levels of oxidized low-density lipoprotein (ox-LDL) have been reported to increase the risk of cardiovascular disease. Furthermore, the anti-oxidative properties of red wine have been invoked to explain the paradox of low cardiovascular mortality, in spite of high-fat intake, in the French population. However, the distribution of ox-LDL-antibodies (ox-LDL-Ab) in the unselected population and its relationship with red wine consumption are unknown. This study was carried out to identify factors associated with ox-LDL-Ab, with particular emphasis on the association with wine consumption., Methods: We performed a cross-sectional study of 551 unselected, community-dwelling older adults (aged 65 to 94 years) living in Dicomano (Italy). Ox-LDL-Ab titre was measured with an ELISA method., Results: In the whole study sample, ox-LDL-Ab increased with age and was unrelated to blood lipids, HbA(1c), and smoking habit. A significant inverse relationship was found between ox-LDL-Ab titre and daily wine intake, which persisted after adjusting for possible confounders or in subgroup analyses of participants who reported to drink wine or were free from significant morbidity., Conclusions: These results suggest that wine affects ox-LDL-Ab titre in older populations, possibly because of its antioxidant properties.

To develop a trial with lymphocyte suicide gene therapy in patients with hematological malignancies, we transduced human T lymphocytes with a retroviral vector (LSN-tk) encoding the herpes simplex virus thymidine kinase (tk) and the neomycin resistance (NeoR) genes. Precise quantification of gene transfer is crucial for any gene therapy protocol, but methods using semiquantitative PCR are inaccurate and subject to variations. Real-time quantitative PCR could be a valid alternative. A TaqMan probe was designed to hybridize with the NeoR gene. The PCR product is 64 nucleotides long and readily quantified by TaqMan probe binding. The analysis was performed soon after transduction and repeated after the selection procedure. This method was more accurate, reproducible, and sensitive than the semiquantitative PCR assay. Accuracy was the same whether the analysis was performed at the highest rate or at the lowest rate of transduction. Additionally we used real-time PCR to monitor the kinetics of enrichment of the transduced cells over the selection time and showed how 7 days of selection are needed. This study precisely quantified the percentages of cells transduced by the retroviral vector and could have major implications in gene therapy studies.

Beta-blockers effectively reduce mortality and recurrent cardiovascular events in patients surviving myocardial infarction. However, these agents are underused in clinical practice, especially in older patients with diabetes mellitus. The current literature shows that beta-blockers are at least as effective in older diabetic patients as in other patients, without major safety concerns for their possible adverse effects. The use of these agents in clinical practice should therefore be encouraged, in order to reduce the burden of cardiovascular mortality and morbidity in older diabetic patients post-infarction.

Objective: We investigated cardiac and vascular remodeling in an unselected older population with either diastolic hypertension (HTN) or isolated systolic hypertension (ISH)., Background: Isolated systolic hypertension accounts for a substantial proportion of hypertension in individuals older than 65 years and is strongly associated with an increased risk of cardiac and cerebrovascular events. The exact mechanisms underlying the increased risk associated with ISH and elevated pulse pressure (PP), in comparison with HTN, have not been extensively investigated., Methods: Community-dwelling residents age >/=65 years in a small town in Italy (Dicomano) were enrolled. Untreated subjects considered in this study included 173 normotensive subjects (blood pressure [BP] <140/90 mm Hg), 95 subjects with HTN (diastolic BP >/=90 mm Hg), and 43 subjects with ISH (BP >/=160/<90 mm Hg). All subjects underwent extensive clinical examination, echocardiography, carotid ultrasonography, and carotid applanation tonometry., Results: Subjects with ISH had higher left ventricular (LV) mass, which was independently related to PP but not to systolic or mean pressures. Both carotid wall cross-sectional area and vascular stiffness were greater in ISH patients than in HTN and normal subjects and were independently related to PP but not to systolic BP. In addition, ISH was associated with a higher prevalence of carotid plaque and more extensive carotid atherosclerosis., Conclusions: In our community-based elderly population, individuals with ISH had higher prevalences of LV hypertrophy and carotid atherosclerosis than subjects with HTN despite lower mean BP. These findings provide potential pathophysiologic mechanisms underlying the associations of ISH and PP with increased risk of cardiovascular morbidity and mortality.

The herpes simplex virus thymidine kinase (HSV-tk) gene conferring ganciclovir (GCV)-specific sensitivity to transduced cells might control Graft-versus-Leukemia (GvL)/Graft-versus-Host Disease (GvHD). Human T lymphocytes were engineered with an LSN-tk retroviral vector encoding tk and neomycin resistance (NeoR) genes. A total of 80 x 10(6) tk(+) lymphocytes were injected intraperitoneally in NOD-SCID mice. Engraftment was evaluated by human CD45(+)/CD3(+) cytofluorimetric analysis and NeoR-based polymerase chain reaction (PCR) on peripheral blood, bone marrow, liver, thymus, and spleen on day +5. After 14 days, GCV (10 mg/kg daily) cytofluorimetric analysis and PCR were repeated (day +19). Immunohistological studies with anti-CD3 monoclonal antibody followed by alkaline phosphatase and monoclonal anti-alkaline phosphatase staining were performed on spleen and liver at the same time points. Human CD45(+)/CD3(+) cells were engrafted in all tissues on day +5 according to cytofluorimetry, immunohistology, and PCR. Lymphocytes "homed" to the white pulp T-cell area and to the red pulp; liver localization is prevalently at the periportal area. After GCV (day +19), cytofluorimetry and immunohistology showed very few CD3(+) cells. PCR identified the transgene in 22% tissue samples (positive only in thymus and spleen). GvHD did not occur in any animal. These data demonstrate elevated doses of human-transduced CD3(+) cells engraft in NOD/SCID mice; after GCV, very few CD3(+) cells can be detected and those that escape treatment can be found in the thymus and in the spleen on day +19. Lack of full response to GCV may account for cases of GvHD in patients receiving tk-transduced T lymphocytes.

Objectives: To determine whether advanced age affects the immediate and long-term results of direct-current external cardioversion (ECV) of atrial fibrillation (AF), the sustained arrhythmia most commonly encountered in older patients., Design: Retrospective analysis of medical records., Setting: Intensive care unit., Participants: Two hundred fifty consecutive patients(age 34-100) with AF who underwent ECV following a standardized protocol in an intensive care unit., Measurements: Immediate efficacy of ECV, defined as recovery of sinus rhythm, and maintenance of sinus rhythm over the follow-up were study outcomes. The univariate and multivariate associations of immediate efficacy of ECV and long-term results with clinical variables were analyzed., Results: At univariate analysis, immediate efficacy of ECV (overall, 91.2%) was lower in older patients and in those with chronic obstructive pulmonary disease, higher for a 3- to 90-day pre-ECV duration of AF than for a duration of 2 days or less or more than 90 days, and independent of underlying cardiac disease, hypertension, diabetes mellitus, previous AF, and left atrial dimension. However, pre-ECV duration of AF was the only multivariate predictor of ECV immediate success. Major complications occurred in only three patients. Of 220 patients discharged in sinus rhythm, 211 were followed up for a mean period +/- standard deviation of 34 +/- 25 months. AF relapsed in 45.5% of them. At multivariate analysis, underlying cardiac disease was the only predictor of the relapse rate, and relapse rate was lower in coronary heart disease than in valvular heart disease, congestive heart failure, or lone AF., Conclusion: Immediate and long-term results of ECV of AF, an effective and safe procedure, are unaffected by age,at least after adjusting for several covariates.

Microsatellites have become one of the most powerful genetic markers in biology. We have used DNA sequencing to characterize a highly variable microsatellite (GAAA) locus in the root-knot nematode Meloidogyne artiellia. The use of microsatellite flanking primers produced four amplification products that are defined as electromorphs, based on conventional length criteria. The sequencing of these four amplification products revealed the presence of new variants in the population due to sequence variability. The sum of electromorphs and sequence polymorphisms resulted in a total of six variants. The high degree of variability in the microsatellite containing region is due not only to variation in the number of tetranucleotide repeats but also to variation (length and site variation) in the flanking regions of the microsatellite. These investigations show that, in spite of the size homoplasy, the variability of the microsatellite flanking sequences of M. artiellia could be used as informative markers for phylogenetic reconstructions.

Cerebral protection during hypothermic circulatory arrest was obtained by combining right carotid perfusion through the axillary artery with selective perfusion of the left common carotid artery in ten patients. We believe that the proposed technique offers several advantages. (1) The axillary artery is generally free from atherosclerosis. (2) It eliminates the risk of retrograde embolization. (3) It provides antegrade perfusion of the true lumen in aortic dissection. (4) Antegrade cerebral perfusion is never interrupted. (5) Bihemispheric perfusion is assured. (6) The risk of air embolysm during carotid cannulation is reduced.

Background: Angiotensin-converting enzyme (ACE) inhibitors prevent decline in physical function in patients with congestive heart failure (CHF). We aimed to see whether ACE inhibitors also prevent reduction in physical performance and in muscle strength in older women who do not have CHF., Methods: We assessed 3-year rates of decline in both knee extensor muscle strength and walking speed in 641 women with hypertension who had participated in the Women's Health and Aging Study. Women were stratified into four groups according to type and duration of antihypertensive drug treatment. 61 had used ACE inhibitors continuously, 133 intermittently, 146 never, and 301 had used other hypertensive drugs either continuously or intermittently., Findings: Participants who had taken ACE inhibitors continuously had a lower mean 3-year decline in muscle strength of -1.0 kg (SE 1.1) compared with -3.7 (0.5) kg in continuous/intermittent users of other antihypertensive drugs (p=0.016) and with -3.9 kg in those who had never used antihypertensives (p=0.026). Muscle strength fell by 3.0 kg in 3 years in both continuous and intermittent users of ACE inhibitors (p=0.096). Mean 3-year decline in walking speed in continuous ACE inhibitor users was -1.7 cm/s compared with -13.6 cm/s in intermittent users of ACE inhibitors (p=0.015), -15.7 cm/s in continuous/intermittent users of other antihypertensive drugs (p=0.002), and -17.9 cm/s in never users of antihypertensive drugs (p=0.001)., Interpretation: ACE inhibitor treatment may halt or slow decline in muscle strength in elderly women with hypertension and without CHF.

This study examines and compares the effect of aerobic and resistance exercise on emotional and physical function among older persons with initially high or low depressive symptomatology. Data are from the Fitness, Arthritis and Seniors Trial, a trial among 439 persons 60 years or older with knee osteoarthritis randomized to health education (control), resistance exercise, or aerobic exercise groups. Depressive symptoms (assessed by the Center for Epidemiologic Studies--Depression scale) and physical function (disability, walking speed, and pain) were assessed at baseline and after 3, 9, and 18 months. Compared with results for the control group, aerobic exercise significantly lowered depressive symptoms over time. No such effect was observed for resistance exercise. The reduction in depressive symptoms with aerobic exercise was found both among the 98 participants with initially high depressive symptomatology and among the 340 participants with initially low depressive symptomatology and was the strongest for the most compliant persons. Aerobic and resistance exercise significantly reduced disability and pain and increased walking speed both, and to an equal extent, in persons with high depressive symptomatology and persons with low depressive symptomatology.

Background: Aspartate aminotransferase (AST) is an enzyme normally confined to the cytoplasm of cells, but released to the exracellular environment upon cell death. Its levels are associated with the severity of experimental gingivitis and the loss of periodontal attachement. The aim of the present study was to investigate the presence and activity levels of AST in peri-implant crevic-ular fluid (PCF) healthy and diseased endosseous implants in order to assess if AST in PCF can be further studied as a possible objective diognostic aid in oral implantology., Methods: Eighty-one fitures from 81 systemically healthy subject were divide into 3 groups, 27 healthy implants (HI), 27 implants with mucositis (MI) and 27 implants affected by periimplantities (PI)according to well-defined clinical and radiographic criteria. PCF was collected by the insertion of a # 40 standardized - point to the base of the crevice or pocket for 30 seconds. AST activity was determined spectrophotometrically at 25' C. The results were expressed as AST Units/ml in PCF., Results: An AST activity was detected in each sample from HI, MI and PI. The mean AST activity in HI was 0.26 f 0.16 U/ml; in MI, 0.38 ± 0.27u/ml; in PI, 0.62 k 0.29 U/ml. ANOVA showed that the deference among HI, MI, and PI was statistically significant at P <0.01level. Post-hoc tests demonstrated that a significant difference in AST activity existed between HI/PI (t = 5.14; P <0.01) and MI/PI (t = 3.09; P <0.01). No statistically significant difference was fond between HI/MI (t = 1.07; P>0.1) AST activity was significantly (P <1) associated with probing depth (r = 0.55), the amount of bone loss (r = 0.60) and bleeding on probing (r = 0.67). When the threshold for a positive AST test was set ≥0.4 U/ml, a sensitivity = 0.81 and a specificity = 0.74 were found in the detection of peri-implantitis; the positive predictive value was 61% and the negative predictive value was 88%., Conclusions: within the limits of this study, our results may suggest the PCF analysis could be further investigated in longitudi-nal studies as a suitable diagnostic strategy in the evaluation of dental implants. J Periodontol2000;71:1151-1157., (© 2000 American Academy of Periodontology.)