Development and characterization of microsatellite loci for Khaya senegalensis (Meliaceae)1.

PREMISE OF THE STUDY: Microsatellite loci were developed to characterize genetic variation and population subdivision in Khaya senegalensis (Desr.) A. Juss. (Meliaceae). • METHODS AND RESULTS: Microsatellite loci were identified from genomic DNA sequences generated using the 454 GS-FLX titanium platform. Primers were designed for 67 tri- and tetranucleotide repeats, of which 20 were selected for 2 multiplexes based on amplification success and band size. Eleven of these loci showed polymorphism in two populations of Khaya senegalensis and amplified in individuals from across the species range. • CONCLUSIONS: These new microsatellite loci will be useful for investigation of the evolutionary and conservation genetics of Khaya senegalensis.

Automatic generation of alignments for 3D QSAR analyses.

Many 3D QSAR methods require the alignment of the molecules in a dataset, which can require a fair amount of manual effort in deciding upon a rational basis for the superposition. This paper describes the use of FBSS, a program for field-based similarity searching in chemical databases, for generating such alignments automatically. The CoMFA and CoMSIA experiments with several literature datasets show that the QSAR models resulting from the FBSS alignments are broadly comparable in predictive performance with the models resulting from manual alignments.

Determinants of tapping speed in normal control subjects and subjects with Parkinson's disease: differing effects of brief and continued practice.

BACKGROUND: Alternate tapping speed is widely used as a measure of bradykinesia in Parkinson's disease (PD). Tapping speed in normal control subjects and factors that might influence tapping speed have not been systematically examined. OBJECTIVE: To examine the effects of age, hand dominance, and gender on tapping speed in normal control subjects and to compare the effects of practice on tapping speed in normal and PD control subjects. METHODS: Tapping speed for three sequential trials in the dominant and nondominant hand was examined in 100 normal control subjects and 60 subjects with PD. The effect of hourly practice over 26 hours (19 trials) was investigated in 14 normal and 24 PD subjects. RESULTS: The speed with which normal subjects alternately tapped two counters was negatively correlated with age, was greater in the dominant hand, was not related to gender, and improved with short-term practice (three trials) and with continued practice over 26 hours. Parkinsonian subjects, in general, tapped more slowly than normal control subjects and more slowly in the more affected arm. Parkinsonian subjects benefited from short-term practice as much as normal control subjects but, unlike normal control subjects, did not improve with continued practice over 26 hours. CONCLUSIONS: Alternate tapping speed is influenced by age, hand dominance, Parkinson's disease, and practice. Subjects with PD do not benefit as much from continued practice as do normal subjects, suggesting some limitation or impairment of procedural (motor) learning in PD.

Prospective blinded study of the relationship between plasma homocysteine and progression of symptomatic peripheral arterial disease.

PURPOSE: An elevated plasma homocysteine level is an established risk factor for atherosclerotic coronary heart disease (CHD), cerebrovascular disease (CVD), and lower extremity occlusive disease (LED). An elevated plasma homocysteine level can be reduced by therapy with folate and vitamins B6 and B12. An accurate evaluation of the role of vitamin therapy requires knowledge of the influence of plasma homocysteine levels on the progression of CHD, CVD, and LED. METHODS: The Homocysteine and Progression of Atherosclerosis Study is a blinded prospective study of the influence of homocysteine and of other atherosclerotic risk factors on the progression of disease in patients with symptomatic CVD, LED, or both. This study is set in a university hospital vascular surgery clinic and the General Clinical Research Center. Consecutive patients with stable symptomatic CVD or LED underwent baseline clinical, laboratory, and vascular laboratory testing for homocysteine and other risk factors and were examined every 6 months. The primary endpoints were ankle brachial pressure index, duplex scan-determined carotid stenosis, and death. The secondary endpoints were the clinical progressions of CHD, LED, and CVD. The hypothesis that was tested was whether the progression of symptomatic CVD or LED was more frequent or more rapid in patients with elevated plasma homocysteine levels. plasma homocysteine levels. RESULTS: After a mean follow-up period of 37 months (range, 1 to 78 months) for deaths from all causes (>14 micromol/L; elevated, 18.6%; normal, 9.4%; P = .022), deaths from cardiovascular disease (elevated, 12.5%; normal, 6.3%; P = .05) and the clinical progression of CHD (highest 20% of homocysteine levels, 80%; lowest 20% of homocysteine levels, 39%; P = .007) were significantly more frequent or more rapid by life-table analysis when the homocysteine levels were elevated. Multivariate Cox proportional hazards regression model showed a significant independent and increasing relationship between the plasma homocysteine levels and the time to death (relative risk for highest one third of homocysteine values, 1.6; 95% confidence interval [CI], 1.04 to 2.56; P = 029; and relative risk for highest one fifth of homocysteine values, 3.13; 95% CI, 1.69 to 6.64; P = .0001). After an adjustment for age, smoking, hypertension, diabetes, cholesterol, and the vascular laboratory progression of CVD or LED, each 1.0 micromol/L increase in the plasma homocysteine levels resulted in a 3.6% increase (95% CI, 0.0% to 6.6%; P = .06) in the risk of death (all causes) at 3 years and a 5.6% increase (95% CI, 2.2% to 8.5%; P = .003) in the risk of death from cardiovascular disease. CONCLUSION: We conclude that elevated plasma homocysteine levels are associated significantly with death, with death from cardiovascular disease, and with the progression of CHD in patients with symptomatic CVD or LED. These results strongly mandate clinical trials of homocysteine-lowering vitamin therapy in such patients.

Evidence for association of HLA-A2 allele with onset age of Alzheimer's disease.

Our earlier studies had suggested a possible association between the HLA-A2 allele and Alzheimer's disease (AD). In the present study we tested the hypothesis that A2 is associated with earlier AD onset. We performed two independent studies: a collaborative study with 111 patients and a confirmatory study with 96 patients. We found similar patterns of reduced age at onset as a function of A2 in both data sets. Overall, A2 was associated with a significant 3-year shift to earlier onset. The effects of A2 and epsilon 4 on age at onset appeared additive. Our results suggest A2, or a closely linked gene, modulates onset age of AD. Association with A2 would suggest an immune/inflammatory response mechanism for AD.

Gender difference in apolipoprotein E-associated risk for familial Alzheimer disease: a possible clue to the higher incidence of Alzheimer disease in women.

Late-onset Alzheimer disease (AD) is associated with the apolipoprotein E (APOE)-epsilon4 allele. In late-onset familial AD, women have a significantly higher risk of developing the disease than do men. The aim of this study was to determine whether the gender difference in familial AD is a function of APOE genotype. We studied 58 late-onset familial AD kindreds. Kaplan-Meier survival analysis was used to assess genotype-specific distributions of age at onset. Odds ratios were estimated by logistic regression with adjustment for age and by conditional logistic regression with stratification on families. All methods detected a significant gender difference for the epsilon4 heterozygous genotype. In women, epsilon4 heterozygotes had higher risk than those without epsilon4; there was no significant difference between epsilon4 heterozygotes and epsilon4 homozygotes. In men, epsilon4 heterozygotes had lower risk than epsilon4 homozygotes; there was not significant difference between epsilon4 heterozygotes and those without epsilon4. A direct comparison of epsilon4 heterozygous men and women revealed a significant twofold increased risk in women. We confirmed these results in 15 autopsy-confirmed AD kindreds from the National Cell Repository at Indiana University Alzheimer Disease Center. These observations are consistent with the increased incidence of familial AD in women and may be a critical clue to the role of gender in the pathogenesis of AD.

Differential permeability of the blood-tumour barrier in intracerebral tumour-bearing rats: antidrug antibody to achieve systemic drug rescue.

The feasibility of utilizing the differential permeability of the blood-tumour barrier to low- vs. high-molecular-weight compounds is demonstrated in a brain tumour model. Nude rats (n = 27) with or without intracerebral tumours received intravenous [3H]methotrexate (M(r) 454), followed 60 min later by antimethotrexate antibody (M(r) 150,000) or nonspecific mouse antibody. Antimethotrexate antibody resulted in 93% binding of serum methotrexate. In contrast, the percentage of antibody-bound methotrexate in brain and intracerebral tumour was only slightly greater than preantibody protein binding. Methotrexate delivery to tumour was significantly greater than to brain adjacent to tumour and normal brain. The percentage delivery of [3H]methotrexate to all areas of brain was similar between animals receiving antimethotrexate antibody and nonspecific antibody. These findings support the theory that a drug rescue method may be developed that may permit the safe administration of increased dosages of chemotherapeutic drugs for the treatment of intracerebral tumours.

Spontaneous echo contrast and hemorheologic abnormalities in cerebrovascular disease.

BACKGROUND AND PURPOSE: Spontaneous echo contrast (SEC) is thought to represent a risk factor for cardioembolic stroke. In vitro studies suggest that SEC results from interaction between red cells and fibrinogen. To better understand the relation between SEC and stroke and to investigate the in vivo genesis of SEC, we examined the relation between SEC, the constituents of the blood, and plasma and serum viscosity in patients with acute stroke or chronic cerebrovascular disease. METHODS: Fifty patients with acute stroke or chronic cerebrovascular disease referred for transesophageal echocardiogram (TEE) were studied by transthoracic echocardiography and TEE. Complete blood count, fibrinogen, albumin, gamma-globulin, and plasma and serum viscosity determinations were made. Left atrial SEC was graded as absent, mild, or marked by means of TEE. RESULTS: SEC was absent in 31 patients, mild in 10 patients, and marked in 9 patients. Higher grade of SEC was associated with a significantly greater percentage of patients with atrial fibrillation and larger left atrial dimension. Atrial fibrillation was present in 23% of the patients in the SEC absent group, 50% of the patients in the mild SEC group, and 78% of the patients in the marked SEC group (P < .01). Left atrial diameter averaged 3.8 +/- 0.6 cm in the SEC absent group, 4.3 +/- 1.1 in the mild SEC group, and 4.9 +/- 0.7 in the marked SEC group (P < .001). Hematocrit, white blood cell count, and platelet count did not differ among the three groups. Fibrinogen, gamma-globulin, plasma viscosity, and serum viscosity values were all significantly higher in the presence of SEC (P < .05). Fibrinogen values were 361 +/- 97 mg/dL in the SEC absent group and 427 +/- 135 mg/dL in the marked SEC group. gamma-Globulin levels were 0.75 +/- 0.23 g/dL in the SEC absent group and 1.06 +/- 0.48 g/dL in the marked SEC group. Both plasma viscosity (1.97 cp) and serum viscosity (1.64 cp) were higher in the marked SEC group than in the SEC absent group (1.77 and 1.50 cp, respectively). CONCLUSIONS: In patients with acute stroke or chronic cerebrovascular disease, the severity of SEC was not related to albumin, hematocrit, white cell count, or platelet count but rather to elevated fibrinogen levels and concomitant increases in both plasma and serum viscosity. Moreover, increasing grade of SEC was associated with significantly increased left atrial diameter and a higher percentage of patients in atrial fibrillation.

Automation of conformational analysis and other molecular modeling calculations.

A software system has been developed for facilitating modeling calculations on large numbers of molecules. Using the system, it is possible to subject one or more molecules to a series of calculations, each requiring use of a different computer program. No user intervention is required: where necessary, output from one program is used automatically as input to the next. Names are assigned to output files automatically and in a systematic manner. As an example, the system can be used to perform a succession of calculations aimed at identifying the major low-energy conformers of each of a set of molecules, starting only from their chemical connectivities. The reliability of the results has been tested by calculations on 40 molecules taken from the Cambridge Structural Database. The observed crystal structure geometry could be found for the majority of these molecules.

Comparative hypolipidemic effects of lovastatin and simvastatin in patients with heterozygous familial hypercholesterolemia.

We have compared the effects of lovastatin and simvastatin on plasma lipoproteins, fibrinogen and urinary mevalonic acid excretion in twenty-three patients with heterozygous familial hypercholesterolemia. After a baseline period patients were randomly assigned to receive lovastatin or simvastatin at doses of 10, 20 and 40 mg twice daily, for a period of 2 months each, and then, after a 4-week wash-out period, all patients received the alternate drug for a similar period of therapy. Both drugs were well-tolerated and no patients were withdrawn due to side effects. Lipid values returned to baseline after discontinuation of therapy and no carry-over effect was observed. Treatment with lovastatin resulted in decreases in LDL cholesterol concentrations from 274 mg/dl at baseline to 211, 192 and 178 mg/dl, respectively, on doses of 20, 40 and 80 mg/day. Treatment with simvastatin reduced concentrations of LDL cholesterol to 194, 168 and 156 mg/dl, respectively, on doses of 20, 40 and 80 mg/day. Concentrations of HDL cholesterol increased on both drugs, but no dose response relationship was apparent. Both drugs reduced the 24-h urinary excretion of mevalonic acid, an intermediate in cholesterol biosynthesis, but the magnitude of decrease was similar with lovastatin and simvastatin. Small, but statistically non-significant decreases in fibrinogen occurred with both drugs. Patients who showed the greatest hypolipidemic effect during treatment with lovastatin also showed an excellent therapeutic response to simvastatin and vice versa. We conclude that, on a milligram per milligram basis, simvastatin is twice as potent as lovastatin in the treatment of familial hypercholesterolemia and that with both drugs, reductions in LDL cholesterol concentrations are accompanied by decreases in the urinary excretion of mevalonic acid.

Hypocholesterolemic effects of mevinolin in patients with heterozygous familial hypercholesterolemia.

We have evaluated the hypolipidemic effects of mevinolin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, the rate-limiting enzyme in cholesterol biosynthesis in 13 patients with heterozygous familial hypercholesterolemia (FH). Patients were maintained on a low-cholesterol diet and received sequentially increasing doses of 5, 10, 20, and 40 mg of mevinolin twice daily for a period of 1 mo on each dose. Plasma concentrations of low density lipoprotein cholesterol decreased by 19.8% on the 5 mg twice daily dose (P less than 0.05 vs. base line), 28.4% on 10 mg of mevinolin twice daily (P less than 0.05 vs. 5 mg twice daily), 35% on 20 mg of mevinolin twice daily (P less than 0.05 vs. 10 mg twice daily), and 37.7% on 40 mg of mevinolin twice daily (not statistically different from 20 mg twice daily). Concentrations of high density lipoprotein cholesterol remained stable on all doses of mevinolin whereas plasma triglyceride levels fell significantly on the 20 mg (-30.7%) and 40 mg (-34.3%) twice daily doses of mevinolin. Mevinolin was well tolerated and all patients completed the study period. Side effects during the period of study were limited to transient insomnia and headaches in two patients, transient increases in alkaline phosphatase in three patients, and a modest but sustained increase in alkaline phosphatase in a fourth patient. These results indicate that mevinolin is an effective hypolipidemic agent in patients with heterozygous FH but that the optimal doses in these patients are greater than those previously reported in normal volunteers. If long-term safety can be satisfactorily established, mevinolin offers considerable promise in the therapy of heterozygous FH.

Acute effects of volcanic ash from Mount Saint Helens on lung function in children.

To evaluate the acute effects of volcanic ash from Mt. St. Helens on the lung function of children, we studied 101 children 8 to 13 yr of age who were attending a 2-wk summer camp for children with diabetes mellitus in an area where about 1.2 cm of ash had fallen after the June 12, 1980, eruption. The outcome variables used were forced vital capacity, forced expiratory volume in one second, their ratio and mean transit time. Total and respirable dust levels were measured using personal sampling pumps. The children were tested on arrival and twice (early morning [A.M.] and late afternoon [P.M.]) every second or third day during the session. A within-day effect was measured by the P.M./A.M. ratio for the lung function variables; a between-day effect was measured by the change in the P.M. measurements over the 2 wk of camp. We found no strong evidence of either a within-day or a between-day effect on lung function, even in a subgroup of children who had preexisting lung disease or symptoms, despite daytime dust/ash levels that usually exceeded the Environmental Protection Agency's significant harm level for particulate matter.

Ciprofibrate in the therapy of type II hypercholesterolemia. A double-blind trial.

The hypolipidemic efficacy of ciprofibrate was evaluated in patients with type II hypercholesterolemia. Patients were randomized to placebo or ciprofibrate (50 mg or 100 mg/day) and, after a 6-week baseline period, received medication for a period of 12 weeks. Blood samples were analyzed every 2 weeks. Twenty patients completed the study (4 on placebo, 7 on 50 mg/day, and 9 on 100 mg/day ciprofibrate). The drug was well tolerated in all patients. Lipid values in the patients on active drug decreased and attained stable values after 4 weeks of treatment. Compared to baseline values, total and LDL cholesterol decreased 11% and 13% on the 50-mg dose whereas HDL increased 8%. Plasma triglyceride fell by 22%. In patients receiving 100 mg ciprofibrate, total and LDL cholesterol fell by 20% (334 leads to 269 mg/dl) and 24% (262 leads to 198 mg/dl), respectively. HDL increased 9.8% (51 leads to 56 mg/dl) and triglyceride decreased by 30% (102 leads to 69 mg/dl). Values in the placebo group remained stable. We conclude that once daily therapy with 100 mg ciprofibrate, is effective in reducing LDL levels in patients with type II hypercholesterolemia (mainly heterozygous FH) and that this decrease is paralleled by small rises in HDL.

Pulmonary function in young children with alpha 1-antitrypsin deficiency: comparison with matched control subjects.

In this paper we report the initial cross-sectional data from a prospective study of pulmonary function in children with moderately severe and severe alpha 1-antitrypsin deficiency. Using a case-control design, our cases were 19 children 3 to 7 years of age with alpha 1-antitrypsin deficiency, Pi phenotype ZZ or SZ. Control subjects were selected from healthy children participating in a study to establish reference values for functional residual capacity and maximal expiratory flow at functional residual capacity, using a 1:1 match for sex, height, age, and weight. We found no significant difference between the cases and their matched control subjects with respect to functional residual capacity and maximal expiratory flow at functional residual capacity. We conclude that through 7 years of age there is no gross impairment in overall pulmonary function in children with moderately severe and severe alpha 1-antitrypsin deficiency, Pi phenotypes ZZ and SZ.

Reference values for functional residual capacity and maximal expiratory flow in young children.

To determine the relation of functional residual capacity and maximal expiratory flow at functional residual capacity to sex, age, height, and weight in healthy young children living in Portland, Oregon, we tested 37 boys and 36 girls using a modified helium-dilution technique and partial expiratory flow-volume curves. Within the age, height, and weight ranges studied, exponential or multiple regression techniques offered no substantial advantage over simple linear regression using height, weight, or age. There were no sex differences for the relationship between either variable and age, height, or weight. These techniques can readily be used in children as young as 3 years of age and may provide a method for studying lung growth and development in early childhood and a way to observe the progression of disease or the effect of treatment in the young child.

Pulmonary function in heterozygotes for alpha,-antitrypsin deficiency: a case-control study.

In this paper we present the initial cross-sectional data from a prospective study of lung aging in heterozygotes for alpha 1-antitrypsin deficiency. Using a case-control design, our cases included 37 heterozygotes for alpha 1-antitrypsin deficiency, protease inhibitor phenotypes MZ and MS, selected because they were parents of children with homozygous alpha 1-antitrypsin deficiency identified in a statewide newborn screening program between 1971 and 1974. All of the heterozygotes were less than 40 yr of age. Our control subjects were selected from a random sample of a working population participating in a longitudinal study of lung aging, using a 2:1 match of control subjects to cases, matching age, sex, ethnic origin, and smoking. Using a respiratory symptom questionnaire, spirometry, and the single-breath N2 test, we found no significant difference between heterozygotes and control subjects in terms of respiratory symptoms or pulmonary function data. We conclude that to 40 yr of age, the heterozygous phenotype (Pi MZ and MS) is not a risk factor for impairment of pulmonary function.

The effect of smoking cessation on pulmonary function: a 30-month follow-up of two smoking cessation clinics.

To obtain further information about the effects of cessation of smoking on pulmonary function, we followed subjects who attended 2 smoking cessation clinics during a period of 30 months. This paper reports the results from 15 persons who succeeded in stopping smoking for the full 30-month period and from 42 who did not succeed for more than one month. Testing included a respiratory questionnaire, spirometry, and the single-breath N2 test. Standardized methods, the same equipment, and the same experienced personnel were used throughout the study. We found that forced vital capacity, one-second forced expiratory volume, closing volume as a percentage of vital capacity, closing capacity as a percentage of total lung capacity, and the slope of the alveolar plateau of the single-breath N2 test all improved significantly in the subjects who stopped smoking. This improvement continued for as long as 6 to 8 months, and then remained stable. There was no sex difference in the response to smoking cessation, nor could we find a threshold of function below which cessation did not result in improvement. On the contrary, those subjects with the greatest impairment initially showed the greatest improvement. Respiratory symptoms virtually disappeared in those who stopped smoking. Subjects who continued to smoke showed an initial improvement in some function tests, probably due to a marked decrease in consumption, but no significant improvement during the whole period. We concluded from this study that cessation of smoking results in definite improvement in pulmonary function, that there is greater improvement in persons who begin with impaired function than in those whose function is initially normal, that respiratory symptoms disappear rapidly.

The effect of smoking cessation and modification on lung function.

The purpose of this study was to obtain more information about the effect on lung function of stopping smoking or of modifying the smoking habit and to determine the time course of change. We followed a group of 75 cigarette smokers who attended a smoking cessation clinic in May 1973, using a respiratory symptom questionnaire, spirometry, closing volumes, and the slope of the alveolar plateau of the single-breath nitrogen test. Subjects were tested before stopping smoking and at 1, 3, 6, and 12 months after the initial testing. We found a significant (P less than 0.05) improvement in closing volume as a percentage of vital capacity and closing capacity as a percentage of total lung capacity at 6 and 12 months and in the slope of the alveolar plateau at 1, 6, and 12 months in those who stopped smoking. There was also a dramatic decrease in respiratory symptoms in those who stopped smoking, a moderate decrease in those who reduced their consumption by at least 25 per cent, and very little change in those who did not appreciably modify their smoking consumption.