Sleeping difficulties and health-related quality of life in Parkinson's disease.

OBJECTIVE: Various sleep-related symptoms occur in Parkinson's disease (PD). Their occurrence with health-related quality of life (HRQL), comorbid sleep disorders, and other comorbidities was studied. METHODS: Altogether, 1447 randomly selected patients with Parkinson's disease, aged 43-89 years, participated in a questionnaire study. A structured questionnaire with 207 items was based on the Basic Nordic Sleep Questionnaire. Questions on demographics, PD, sleep disorders, and comorbidities were included. RESULTS: The response rate was 59.0%, and of these, 80% had answered to all questions that were used in the analyses (N=684). Occurrence of long sleep was found in 26.2% of the subjects, short sleep in 32.5%, poor sleep in 21.2%, sleep deprivation in 33.8%, disrupted sleep in 47.4%, and difficulties to fall asleep in 12.2%, respectively. Poor self-rated health and poor quality of life occurred in 44.4% and in 43.3% of all participants. In the logistic regression, age and gender differentially predicted long sleep and sleep deprivation, such that older age and being male were positively associated with long sleep but negatively associated with the report of sleep deprivation. Depression, subjective negative stress, and fatigue occurred with long sleep. On the other hand, poor sleep and excessive daytime sleepiness occurred with short sleep and sleep deprivation. CONCLUSIONS: The sleep difficulties in PD are frequent. The long sleeping patients have depression, stress, and fatigue.

Economic burden and quality of life impairment increase with severity of PD.

PURPOSE: To define the interrelationship between cost-of-illness, quality of life (QoL) and Parkinson's disease (PD) severity in a common patient management setting in Finland.Scope. Two hundred and sixty consecutive outpatients with idiopathic PD participated. UPDRS, motor fluctuations, QoL, and the use of health care resources were measured. Direct and indirect costs were calculated. CONCLUSIONS: There is a strong relationship between QoL or cost-of-illness on the one hand, and severity of PD on the other. Treatment policies capable of reducing or delaying motor fluctuations would be expected to increase QoL and reduce some of the economic burden of PD.

Simultaneous quantification of human glandular kallikrein 2 and prostate-specific antigen mRNAs in peripheral blood from prostate cancer patients.

We present a multiplexed and internally calibrated quantitative reverse transcription-PCR (QRT-PCR) assay to detect human glandular kallikrein 2 (hK2) and prostate-specific antigen (PSA) transcripts in blood samples from healthy subjects and prostate cancer (PC) patients. The assay detected 50 copies of hK2 and PSA mRNA, and 1 PSA- and 10 hK2-expressing LNCaP cells in the presence of 2.5 x 10(6) PSA- and hK2-negative cells. In PC patients, 20 of 25 and 19 of 25 gave detectable PSA and hK2 mRNAs, respectively. Number of hK2 mRNA copies was significantly higher than that of PSA mRNA copies in patients with biochemically progressive (P = 0.02) PC, and with locally advanced and metastasized (P = 0.004) PC. Patients with rapidly progressive and hormone refractory PC gave detectable hK2 mRNA only in 2 of 8 and PSA mRNA in 3 of 8 patients. Neither PSA nor hK2 mRNAs were detected in 16 healthy subjects. PSA and hK2 discriminated PC patients with biochemically progressive and advanced disease from the controls and from the aggressive distant metastatic disease. The assay provides a reliable quantification of the number of hK2 and PSA mRNA copies, allows to discriminate PC cases from healthy subjects, and offers a tool for further studies on molecular staging of PC.

Dual-label detection of amplified products in quantitative RT-PCR assay using lanthanide-labeled probes.

Quantitative RT-PCR (QRT-PCR) enables the sensitive and specific detection of mRNA with a small copy number. We used the QRT-PCR method and dual-label analysis of amplification products for the detection of prostate-specific antigen (PSA) mRNA. The QRT-PCR assay employed a PSA-like internal standard (IS) mRNA, which was used to quantify the PSA mRNA copies and to control the variations during the whole assay procedure from the RNA extraction to the detection of QRT-PCR amplification products by hybridization assay. After co-amplification, the PSA and IS products were detected in a microplate using Eu3+ chelate-labeled PSA and Tb3+ chelate-labeled IS hybridization probes. The detection probes allowed the simultaneous and dual-label detection of PSA and IS products in the same microtiter well. Compared to the single-label assay, the dual-label detection improved the within- and between-assay CV% from 21.7 to 7.5 and from 36.0 to 30.3, respectively. The between- and within-assay variation of the dual-label assay was further studied using PSA-producing LNCaP cells. The cells were found to express 980 +/- 170 (mean +/- SD) copies of PSA-mRNA with the within-assay CV% of 17.7 and 890 +/- 220 (mean +/- SD) copies of PSA-mRNA with the between-assay CV% of 25.0. The methodology developed may help in future studies to obtain reliable quantification of PSA mRNA generated by circulating prostate cancer cells.

Time-resolved fluorometry in end-point and real-time PCR quantification of nucleic acids.

Two time-resolved fluorescence-based methods for nucleic acid quantification are described and their results are compared. Both methods use an exogenous internal standard to eliminate errors arising from different steps of the assay. The first method is a competitive end-point assay, where the standard competes for the same primers with the actual target sequence, prostate-specific antigen (PSA) cDNA. The standard and target are quantified in a dual-label plate hybridization with lanthanide-labelled probes after a fixed number of PCR cycles. The second method is based on real-time monitoring of PCR and on the use of a novel homogeneous signal generation principle that relies on the use of a 5'-->3' exonucleolytic DNA polymerase and a probe labelled with an environment sensitive, stable and fluorescent lanthanide chelate. In this assay, a non-competitive, exogenous internal standard is used. Both assays have a wide linear range (50-5 x 10(6) and 10-5 x 10(7) input PSA cDNA molecules for the end-point and real-time assays, respectively) and there is a strong correlation between the results obtained with the two assays (r = 1.0). Being somewhat faster to perform, the real-time format is better suited for assays that require high throughput.

Hippocampal and temporal lobe atrophy and age-related decline in memory.

OBJECTIVES: To evaluate the relationship of memory decline that accompanies aging with structural changes in the medial temporal lobe, in healthy middle-aged and older subjects. MATERIAL AND METHODS: A sample of 35 neurologically non-diseased subjects, between 55 and 70 years of age, were examined in a 5-year follow-up study. Neuropsychological investigation included tests of learning, verbal memory, and visual memory. MRI was performed with a superconducting MRI system operating at 1.0 T, using coronal slices of T1-weighted images. Medial temporal lobe atrophy was rated separately in the neocortical, entorhinal and hippocampal regions. RESULTS: We did not find any statistically significant relationship between mild hippocampal or temporal atrophy and memory test performance. Nor did the longitudinal decline in memory show a relationship with temporal lobe atrophy. CONCLUSIONS: The main outcome of our study was that age-related memory decline was not related to mild temporal lobe atrophy in healthy subjects without mild cognitive impairment. There could be other factors influencing memory functions besides age-related structural changes in temporal lobes.

A new label technology for the detection of specific polymerase chain reaction products in a closed tube.

A novel signal generation principle suitable for real time and end-point detection of specific PCR products in a closed tube is described. Linear DNA probes were labeled at their 5'-ends with a stable, fluorescent terbium chelate. The fluorescence intensity of this chelate is lower when it is coupled to single-stranded DNA than when the chelate is free in solution. The synthesized probes were used in the real time monitoring of PCR using a prototype instrument that consisted of a fluorometer coupled to a thermal cycler. When the probe anneals to a complementary target amplicon, the 5'-->3' exonucleolytic activity of DNA polymerase detaches the label from the probe. This results in an enhanced terbium fluorescence signal. Since terbium has a long excited state lifetime, its fluorescence can be measured in a time-resolved manner, which results in a low background fluorescence and a 1000-fold signal amplification. The detection method is quantitative over an extremely wide linear range (at least 10-10(7)initial template molecules). The label strategy can easily be combined with existing label technologies, such as TaqMan 5'-exonuclease assays, in order to carry out multiplex assays that do not suffer from overlapping emission peaks of the fluorophores.

Cardiovascular diseases, health status, brain imaging findings and neuropsychological functioning in neurologically healthy elderly individuals.

The aim of our study was to evaluate the relationship between health-related factors, brain imaging findings and cognitive functioning. We examined 113 neurologically healthy subjects from 55 to 85 years of age. Health-related variables included a clinical health evaluation, cardiovascular diseases, and other systemic diseases. The presence of white matter changes and cerebral and peripheral atrophy were obtained with magnetic resonance imaging. Neuropsychological tests measuring verbal memory, visual memory, intellectual and language functions, visuoconstructional functions, flexibility, and speed and attention were administered. Results showed that overall health status was not related to cognition. Subjects, who had both arterial hypertension and white matter changes had difficulties in flexibility. Cardiac failure and white matter changes were related to impairment in visuoconstructional functions, flexibility and attention. Significant speed and attention deficits were observed in subjects with cardiac failure and central atrophy. In conclusion, this study verifies the relationship between hypertension, white matter changes and cognitive functions. We found also specific patterns in relation with cardiac failure, brain imaging findings and cognitive functioning, the most vulnerable domains were visuoconstructional functions, flexibility and attention.

Heterogeneity of cognitive profiles in aging: successful aging, normal aging, and individuals at risk for cognitive decline.

Neuropsychological clinical decision-making is complicated by the fact that variability in test performance increases with advancing age. This research explores the presence of homogeneous subgroups in 120 neurologically healthy individuals, from 55 to 85 years of age. Subjects at risk for dementing diseases were diagnosed as Aging-Associated Cognitive Decline (AACD) and Mild Cognitive Impairment (MCI). Cluster analysis was applied on 11 neuropsychological variables assessing logical memory immediate recall and retention percentage, visual memory immediate recall and retention, conceptual thinking, naming, verbal fluency, constructional functions, motor speed, flexibility and finger tapping. Five clusters were extracted, one representing cognitively successfully aged, and two consisting of individuals with normal or average level of performance. One cluster was characterized by older subjects with difficulties in visual memory, visuoconstructional functions, and speed and attention, most of the younger subjects in the same cluster had a diagnosis of AACD or MCI. The fifth cluster represented individuals at risk for dementing diseases; most of them were diagnosed having AACD and more than half had a diagnosis of MCI. Age, activity and intellectual levels, and to a lesser degree education, were significantly related to the cluster solution. The present findings caution against treating samples of elderly individuals as homogeneous.

Quantitative reverse transcription-PCR assay with an internal standard for the detection of prostate-specific antigen mRNA.

BACKGROUND: Circulating prostate cells can be detected with a reverse transcription-PCR (RT-PCR) assay for prostate-specific antigen (PSA) mRNA. We have developed a new quantitative RT-PCR method for measuring PSA mRNA. METHODS: The method uses a PSA-like internal standard (IS) mRNA that is added into the sample at the beginning of the RNA extraction and coamplified by RT-PCR with the PSA in the sample. After PCR amplification, the IS and PSA products are selectively detected by hybridization in a microtitration plate using probes labeled with fluorescent europium chelates. RESULTS: The method was validated with PSA and IS mRNAs and PSA-expressing cells to obtain a detection limit of 50 PSA mRNA copies (i.e., signal 2 times the mean of zero signal), linearity up to 10(6) copies, and detection of a single PSA-expressing cell. In preliminary evaluations, 60% (n = 10) of the prostate cancer patients with skeletal metastases gave results above the detection limit (500 PSA mRNA copies in 5 mL of blood). The total number of PSA copies ranged from 900 +/- 200 to 44 100 +/- 4900 (mean +/- SD) in the samples, corresponding to approximately 1-100 PSA-expressing cells in 5 mL of blood. In the controls (n = 34), none of the healthy females and 2 of 19 healthy males had detectable PSA mRNA [700 +/- 100 and 2000 +/- 900 (mean +/- SD) PSA mRNA copies in 5 mL of blood for the 2 males]. CONCLUSIONS: The assay provides sensitive and quantitative detection of PSA mRNA expression from blood samples and can be used to establish the clinically significant number of PSA mRNA copies in prostate cancer.

A dual-label immunofluorometric assay for human osteocalcin.

Circulating human osteocalcin (hOC) has been shown to be comprised of two main forms: the intact 1-49 form and the proteolytic N-terminal midfragment (N-mid) spanning amino acid residues 1-43 or 1-44. We used three monoclonal antibodies (MAbs) raised against hOC and bovine osteocalcin in developing a dual-label assay for the simultaneous measurement of the proportions of the intact and N-mid forms in serum samples. The assay is based on time-resolved fluorescence utilizing differently labeled trace MAbs. Biotinylated MAb 2H9 is used as a capture antibody for both the intact hOC and the N-mid. Tracer MAb 6F9 labeled with a Europium (III)-chelate binds to the intact the N-mid and the intact hOC, whereas tracer MAb 3G8 labeled with a Terbium (III)-chelate binds to the intact hOC only. The simultaneous binding of the antibodies was tested by comparing full-length hOC purified from human bone and hOC shortened from the C terminus by four amino acid residues with carboxypeptidase Y. Serum hOC measurements with the dual-label assay were in agreement with the corresponding single-label assays (r = 0.96 for intact + N-mid assay and r = 0.81 for intact assays, n = 91). The lower correlation between the intact assays was attributable to proteolytic susceptibility of the intact form due to one additional freezing and thawing cycle in carrying out the dual-label assay. As measured with the dual-label assay, the levels (mean +/- SD) of serum intact + N-mid OC were 6.2 +/- 2.1 ng/ml in the premenopausal group (n = 44), 13.9 +/- 4.9 ng/ml in the postmenopausal group without hormone replacement therapy (HRT; n = 13), and 7.5 +/- 3.4 ng/ml in the postmenopausal group with HRT (n = 13). The levels of intact hOC in the same groups were 4.8 +/- 1.4 ng/ml, 9.8 +/- 2.9 ng/ml, and 5.3 +/- 2.1 ng/ml, respectively. Whether the main forms of OC or their relative proportions in serum can be used for predicting bone diseases or for monitoring the progression and management of diseases awaits further investigations.

Differences in neuropsychological functioning associated with age, education, neurological status, and magnetic resonance imaging findings in neurologically healthy elderly individuals.

In a cross-sectional study, a sample of 113 individuals, 55 to 85 years old, without any neurological diseases was investigated. The study provides information on differences associated with age, education, and gender, and in relation to neurological status, magnetic resonance imaging, and cognitive functioning. Differences between age groups were shown in memory, constructional, and language functions, and especially in tests related to speed and attention. Education was related to most of the cognitive functions, but especially to verbal intellectual functions, visual and logical memory, language functions, and calculation. Gender differences were found in finger tapping, constructional functions, and verbal intellectual functions. Primitive reflexes showed a tendency to correlate with comprehension and memory of sentences. Extrapyramidal signs were related to psychomotor speed, and attention, verbal fluency, and set shifting together with intellectual functions and learning. Central atrophy on magnetic resonance imaging was related to memory functions in those 65 and 70 years of age, whereas in the oldest age groups immediate recall was associated with the severity of lesions.

MRI of the brain in neurologically healthy middle-aged and elderly individuals.

Our purpose was to document the MRI appearances of the brain in healthy middle-aged to elderly subjects. T2- and proton density-weighted axial slices were obtained in 61 volunteers, 30-86 years of age. After visual inspection, signal intensities of brain structures were measured on T2-weighted images. Age-related changes became increasingly apparent after age 50. The main findings were that signal intensity of the white matter increased concomitantly with widening of the cerebrospinal fluid spaces; that basal ganglia remained stable; that high-signal foci in white matter increased in number and size after the age of 50 years; that periventricular high-signal foci were constant after the age of 65 years. Our visual impression of a decrease in signal intensity of the central grey matter with age seems to be mistaken. Pathological processes should be suspected if periventricular foci are found in middle-aged or young subjects.

White matter hyperintensities on MRI in the neurologically nondiseased elderly. Analysis of cohorts of consecutive subjects aged 55 to 85 years living at home.

BACKGROUND AND PURPOSE: We undertook this study to evaluate the frequency and risk factors of white matter hyperintensities seen on T2-weighted MR imaging. We examined cohorts of neurologically nondiseased elderly subjects participating in a general-community study, the Helsinki (Finland) Aging Brain Study. Cohorts of consecutive subjects aged 55, 60, 65, 70, 75, 80, and 85 years (n = 20, 18, 20, 18, 19, 18, and 15, respectively; total, n = 128) were divided into a young-old (age < 75 years, n = 76) group and an old-old (age > or = 75 years, n = 52) group. METHODS: Frequency of hyperintensities seen on T2-weighted axial and coronal MR images (0.02 T) was rated using a four-point scale in periventricular and centrum semiovale areas. RESULTS: The majority of the subjects showed only mild white matter hyperintensities, which were more frequent in the periventricular areas. Age was the most important factor to explain the presence of hyperintensities. A logistic regression analysis related periventricular hyperintensities in the entire group to central atrophy (odds ratio [OR], 4.7; 95% confidence interval [CI], 1.7 to 12.9) and silent infarcts (OR, 5.6; 95% CI, 1.0 to 19.8); among the young-old, hyperintensities related to diabetes (OR, 17.0; 95% CI, 1.9 to 154.2) and central atrophy (OR, 14.7; 95% CI, 3.5 to 61.8). Centrum semiovale hyperintensities related in the entire group to cardiac arrhythmia (OR, 4.0; 95% CI, 1.0 to 15.5), central atrophy (OR, 3.9; 95% CI, 1.2 to 12.4), and silent infarcts (OR, 3.6; 95% CI, 1.0 to 12.5). CONCLUSIONS: These mild white matter hyperintensities in the neurologically nondiseased elderly related especially to age and also to concomitant silent infarcts, atrophy, and some vascular risk factors. The known factors, however, explained only part of the variation. The young-old and old-old groups showed different associations. In contrast to former assumptions, the presence of white matter hyperintensities among the aged is likely to be linked to other as yet unidentified age-related factors.

Use of computerized visual performance test in assessing day-time vigilance in patients with sleep apneas and restless sleep.

We have studied day-time vigilance in 31 patients (median age 49 years) with suspected sleep disorders using a new visual reaction time and performance test. The findings in the day-time vigilance test were compared with the number of desaturation events and movement arousals measured with a sensitive movement detector in the night-time. In our statistical model the high number of desaturations correlated with a high dispersion in reaction-times. The squared multiple r was 0.465 in a model where the dispersion of reaction times was the dependent variable and the number of desaturations, duration of quiet sleep and the mode of oxygen saturation were independent variables. A high amount of body movements (movement arousals, duration less than 5 seconds) correlated with gradual deterioration in the performance test. The squared multiple r was 0.447 in a model where the regression coefficient of reaction times was the dependent variable and active sleep and number of body movements less than 5 seconds in duration were the independent variables. Frequent arousals in apnoeic patients are observed in hyper-excitable responders and are known to cause sleep deprivation and hypersomnia. Our findings in desaturating patients indicate that in those with a low chemoreceptor response to hypoxia the failure in day-time regulation of vigilance may differ from the failure associated with sleep-deprivation.

The normal brain stem from infancy to old age. A morphometric MRI study.

Our purpose was to develop a method of measuring the size of the brain stem by routine MRI and to determine brain stem dimensions in a normal population. We examined 174 subjects, aged 4 months to 86 years, with no known brain disease. Sagittal midline diameters of the mesencephalon, pons and medulla oblongata were measured on sagittal T1-weighted images, coronal diameters from axial T2-weighted images. The adult midsagittal diameter of the mesencephalon was reached at the age of 6 years, and decreased slightly after 45-50 years. Pontine dimensions increased until the age of 20 years and did not subsequently decrease. The midsagittal and midcoronal diameters of the medulla oblongata stopped increasing at the ages of 6 and 8 years, respectively. Minimal reduction in the midsagittal diameter occurs after 50 years. Normal ranges for each dimension were recorded. Knowledge of the normal variation in size of the brain stem can be helpful in the investigation of neurodegenerative diseases. The method described is rapid and needs no additional hard- or software. An additional finding was an increase in large vermian sulci in subjects over 50 years of age.

White matter changes in healthy elderly persons correlate with attention and speed of mental processing.

OBJECTIVE: To evaluate the association between white matter changes (leukoaraiosis [LA]) seen on magnetic resonance imaging and cognitive functions. DESIGN: Survey of cohorts of neurologically healthy elderly subjects derived consecutively from a population-based random sample. SETTING: General community, the Helsinki (Finland) Aging Brain Study. SUBJECTS: Cohorts of neurologically healthy subjects aged 55, 60, 65, 70, 75, 80, and 85 years (n = 20, 18, 19, 18, 17, 17 and 11 subjects, respectively; total N = 120). MEASURES: Leukoaraiosis was rated in the periventricular areas (0 to 24) and the centrum semiovale (0 to 24); also, a total LA score was obtained (0 to 48). The neuropsychological test battery covered memory, verbal intellectual and constructional functions, language, speed and attention, and speed of mental processing, as well as simple psychomotor speed. RESULTS: Low age-related LA scores and deterioration of cognitive functions were obtained in the normal subjects. When controlling for age, we found that speed and attention, together with the speed of mental processing measured by the Trail Making A and the Stroop tests, correlated with the total LA score. However, there was wide variation between subjects. Comparing groups with and without LA proved the association of LA with Trail Making A time, Stroop test result (words/time and difference/time), and the compound score of speed and attention. Presence of periventricular LA was especially related to speed of mental processing. CONCLUSION: Leukoaraiosis could explain some of the intellectual impairment in the elderly, especially that of slowing of distinct motor and attentional functions, as well as slowing of mental processing. Mild LA in normal aged subjects could also signal brain at risk for further cognitive impairment.

Magnetic resonance imaging findings associated with cardiac arrest.

BACKGROUND AND PURPOSE: The frequency and prognostic significance of neuroradiological findings after cardiac arrest are unknown. Using healthy volunteers as control subjects, we studied the magnetic resonance imaging (MRI) findings associated with cardiac arrest, adjusted for confounding factors. METHODS: The presence of cerebral infarcts, leukoaraiosis, atrophy, and edema on ultra-low-field MRI was assessed in 88 community volunteers and 52 cardiac arrest survivors enrolled in a placebo-controlled, randomized, double-blind trial of nimodipine in out-of-hospital ventricular fibrillation. RESULTS: Cardiac arrest was an independent risk factor for the presence of infarcts in a logistic regression model adjusted for age, sex, and history of myocardial infarction, stroke, coronary heart disease, cardiac failure, and hypertension (odds ratio, 3.6; 95% confidence interval, 1.3 to 9.9; P = .01). Leukoaraiosis was associated with increasing age but not with cardiac arrest. Adjusted for age, the delay of advanced life support had an inverse correlation with the degree of atrophy in placebo-treated patients (r = -.62, P < .0001) but not in patients treated with nimodipine (r = -.10, P = .43). Lack of age-related atrophy, possibly implicating the presence of brain edema, predicted poor outcome after cardiac arrest (odds ratio, 4.6; 95% confidence interval, 1.4 to 15.8; P = .01). CONCLUSIONS: Cardiac arrest was associated with deep cerebral infarcts but not with leukoaraiosis. MRI findings did not predict the functional outcome at 1 year. Nimodipine treatment had no significant effect on the MRI findings, but delayed resuscitation was associated with probable brain edema only in placebo-treated patients.