Development and validation of a bioanalytical method for quantification of LNA-i-miR-221, a 13-mer oligonucleotide, in rat plasma using LC-MS/MS.

LNA-i-miR-221, a 13-mer oligonucleotide, is a new miR-221 inhibitor that could be used as a novel drug for multiple myeloma. Herein, an ion-pair reversed phase liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantification of LNA-i-miR-221 in rat plasma. Plasma samples were prepared with an initial phenol/chloroform/isoamyl alcohol liquid-liquid extraction followed by a solid phase extraction. Chromatographic separation was performed with a gradient system on a HALO C18 column using hexafluoro-2-propanol/triethylamine buffer and methanol as mobile phase at a flow rate of 0.4 mL/min. Under these conditions LNA-i-miR-221 and the analogue internal standard are co-eluted at 1.2 min. The detection was carried out in multiple reaction monitoring (MRM) mode using a negative electrospray ionization (ESI) interface. The assay showed a good linearity within the calibration range 10-10000 ng/mL. The precision, accuracy, and recovery values were found to be <15% (<20% at LLOQ), 100 ±â€¯15%, and 97.6-103.7%, respectively. This method was successfully applied to measure the concentrations of LNA-i-miR-221 in plasma samples following the intravenous administration during a 4-week toxicity study in rats.

Haplotypes in cathechol-O-methyltransferase gene confer increased risk for psychosis in Alzheimer disease.

BACKGROUND: The gene encoding catechol-O-methyltransferase (COMT) has been suggested as a candidate for Alzheimer-related psychosis (AD-P) susceptibility, and an association between AD-P and a functional valine to methionine polymorphism has been reported. OBJECTIVE: The aim of this study was to assess the genetic contribution of other COMT variants to the risk of AD-P. METHODS: Two hundred and forty-six AD patients underwent clinical and neuropsychological examination as well as an evaluation of behavioural and psychiatric disturbances. They were subsequently divided into two subgroups according to the presence (AD-P) or the absence (AD-nP) of psychotic symptoms. Four single-nucleotide polymorphisms (SNPs) within COMT gene were evaluated, i.e. rs737865, rs737864, intron 1 C2754delC, and the well-known valine/methionine variant (rs4680). Analyses were performed on the single locus and pairwise disequilibrium of loci, and multi-locus haplotype. RESULTS: The individual SNP analysis confirmed an association for the valine/methionine variant with AD-P. Haplotype analyses revealed that the alleles at four loci (rs737865, rs737864, intron 1 C2754delC, rs4680) interacted to create the risk of psychosis in AD, as A-C-C-G haplotype (OR=2.08, 95% CI=1.02-4.27, P=0.044) and G-C-delC-G haplotype (OR=2.54, 95% CI=1.32-4.90, P=0.006) in respect to the most common and not-at-risk A-C-C-A haplotype which was significantly overrepresented in AD-P. CONCLUSIONS: The present findings provide evidence of COMT genetic variations' role in the susceptibility to AD-related psychosis. The observation of a haplotype effect of different polymorphisms within the COMT gene puts emphasis on the usefulness of haplotype analysis in better defining individualized genetic risk profiles in AD.

The Italian version of the pain assessment in advanced dementia (PAINAD) scale.

Pain is an unpleasant sensory and emotional experience. It's recognized to be modified by individual memory, expectation, and emotion. The most accurate evidence of pain and its intensity is based on patient's description and self-report. One of the main problems in assessing pain in dementia concerns with the impairment of communication and memory. Unfortunately, the most used tools to evaluate pain have been developed for normal aging people, requiring verbal and cognitive skills. Therefore, proper instruments are urged to be developed, tested, and validated to assess pain in a cognitively impaired population. The purpose of this study was to assess the validity of the PAINAD in the Italian version as a reliable tool for measuring pain in demented people.

Genetic correlates of behavioral endophenotypes in Alzheimer disease: role of COMT, 5-HTTLPR and APOE polymorphisms.

Several studies have been conducted to understand the genetic correlates of Alzheimer disease (AD)-related behavioral and psychological symptoms in dementia (BPSD). However, given that BPSD rarely occur in isolation, it has been suggested that targeting BPSD individually is too narrow of an approach if one wants to accurately define all the associated risk factors. To date, we know of no work on genetic polymorphisms related to behavioral endophenotypes in AD. The present study sought to evaluate the relationship between such behavioral endophenotypes in AD and genetic variations in dopamine- or serotonin-related genes, such as catechol-O-methyltransferase (COMT) or 5-HTT gene-linked promoter region (5-HTTLPR), and apolipoprotein E (APOE). Among 232 AD patients who underwent clinical and neuropsychological examination, a behavioral and psychiatric evaluation, and genotyping at COMT, 5-HTTPLR, and APOE; 66.4% showed more than one behavioral symptom. By Principal Component Analysis of Neuropsychiatric Inventory (NPI) symptoms four endophenotypes were identified, these were termed "psychosis", "moods", "apathy", and "frontal". Modeling NPI symptom-endophenotype-genotype relationships, and taking into account possible confounds (i.e. demographic characteristics, comorbidities, concomitant pharmacological treatments, and disease severity) by latent variable models, COMT and 5-HTTLPR genetic variations correlated with "frontal" and "psychosis" endophenotypes. APOE genotype did not correlate with any endophenotype. These findings suggest that the possibility of identifying distinct phenotypes on a genetic basis among AD patients exists, and suggest that clustering of BPSD into endophenotypes might provide a new strategy for guiding future research on this issue.

Progressive neuronal and motor dysfunction in mice overexpressing the serine protease inhibitor protease nexin-1 in postmitotic neurons.

Perturbation of the homeostasis between proteases and their inhibitors has been associated with lesion-induced or degenerative neuronal changes. Protease nexin-1 (PN-1), a secreted serine protease inhibitor, is constitutively expressed in distinct neuronal cell populations of the adult CNS. In an earlier study we showed that transgenic mice with ectopic or increased expression of PN-1 in postnatal neurons have altered synaptic transmission. Here these mice are used to examine the impact of an extracellular proteolytic imbalance on long-term neuronal function. These mice develop disturbances in motor behavior from 12 weeks on, with some of the histopathological changes described in early stages of human motor neuron disease, and neurogenic muscle atrophy in old age. In addition, sensorimotor integration, measured by epicranial multichannel recording of sensory evoked potentials, is impaired. Our results suggest that axonal dysfunction rather than cell death underlies these phenotypes. In particular, long projecting neurons, namely cortical layer V pyramidal and spinal motor neurons, show an age-dependent vulnerability to PN-1 overexpression. These mice can serve to study early stages of in vivo neuronal dysfunction not yet associated with cell loss.

One-year mortality in elderly stable patients with COPD.

A retrospective study was performed to evaluate the risks of one-year mortality in very old hospitalized patients including those suffering from chronic obstructive pulmonary disease (COPD). Six hundred and fifty-eight disabled patients (M = 194, mean age 79.2 +/- 7.4 years) consecutively admitted to and discharged from a Geriatric Evaluation and Rehabilitation Unit (GERU) after a comprehensive rehabilitation program were studied and divided into two groups: COPD (n = 337, 51%) and non-COPD (n = 321, 49%). Multidimensional evaluation including information on demographics, cognitive status [Mini Mental State Examination (MMSE)], physical health [number of diseases, Greenfield's Individual Disease Severity (IDS), and number of drugs used], functional disability [Basic Activity of Daily Living (BADL), Tinetti scale, and Physical Performance Test (PPT)], and nutritional status [Prognostic Nutritional Index (PNI)] were assessed at admission. Survival rate was assessed over a 1-year period following discharge. COPD patients mainly differed from non-COPD in terms of older age, smoking habit, number of associated diseases and drugs used. Aggregating the IDS 2-3-4 COPD classes (symptoms + functional impairment), the risk of one-year mortality was double that of the IDS 1 COPD class (symptoms only) and of non-COPD subjects (IDS 0 class) after adjusting for age, sex, disability, malnutrition, and comorbility. Moreover, IDS 2-3-4 COPD patients suffering from cor pulmonale (CP) had a fourfold 1-year risk of mortality in comparison with the IDS 1 COPD group after adjusting for the same covariates. Hospitalized stable very old COPD patients presenting functional impairment have a higher 1-year risk of mortality than only symptomatic COPD or non-COPD subjects. The presence of cor pulmonale with COPD further increases this risk.

Serum cholesterol levels as a measure of frailty in elderly patients.

The authors evaluated the association between serum cholesterol levels and social, clinical, and functional characteristics in 637 elderly hospitalized patients (mean age = 79.1 years, range = 65-97) from the Geriatric Evaluation and Rehabilitation Unit (GERU) at P. Richiedei Hospital in Gussago, Brescia (Italy). Patients consecutively admitted to the GERU during an 18-month period underwent a multidimensional evaluation including information on demographics, cognitive status, physical health (number of chronic diseases and administered drugs), functional disability, and nutritional status. Mean cholesterol levels were significantly lower in men; persons living with others; older individuals; and individuals with cognitive impairment, poorer somatic health, higher disability, and a higher level of malnutrition. Lower serum cholesterol levels may be considered an independent hematologic marker of frailty in elderly hospitalized patients.

Venlafaxine-induced reset osmostat syndrome: case of a 79-year-old depressed woman.

The presence of hyponatremia, especially in a frail and very old patient, is associated with a greater morbidity and mortality rate. We report the case of a depressed 79-year-old woman who was treated with venlafaxine, in whom a drug-induced hyponatremia occurred in the absence of other possible causes. The case is discussed in the context of the multipotential factors that induce hyponatremia, with particular attention to the geriatric patient.

Good nutritional oral intake is associated with equal survival in demented and nondemented very old patients.

OBJECTIVE: To evaluate the association of oral food intake with survival in very old demented nursing home patients. DESIGN: A prospective cohort study. SETTING: A nursing home in northern Italy. MEASUREMENTS: Anthropometric and laboratory nutritional indicators and nutrient intake were assessed in 33 demented (age 85.7 +/- 5.7 years) and 25 nondemented (age 84.9 +/- 5.7 years) patients. Mortality data were collected over a 28-month follow-up period. Association of survival with dementia was estimated by Kaplan-Meyer analysis and multivariate Cox proportional hazard models. RESULTS: Nutrient intake and nutritional status were good compared with data in the literature and were similar in demented and nondemented patients, except for smaller triceps skinfold thickness in the demented. The cumulative annual death rate was 0.23 deaths per subject per year, similar in the demented (0.23) and the nondemented (0.22). Unadjusted survival by Kaplan-Meyer analysis was similar in the two groups, and correction for-age, gender, cognition, triceps skinfold thickness, and number of drugs in a Cox model did not alter the relationship. CONCLUSIONS: Dementia developing in very old age is not necessarily associated with malnutrition and decreased life expectancy.

Association of chronic non-steroidal anti-inflammatory drugs use and cognitive decline in non-demented elderly patients admitted to a geriatric evaluation and rehabilitation unit.

In a geriatric evaluation and rehabilitation unit (GERU), 258 elderly patients (M: 71, F: 187; mean age 77.4 +/- 7.5) scoring 22 or more at Mini-Mental State Examination (MMSE) consecutively admitted were assessed in order to evaluate the effects of non-steroidal anti-inflammatory drugs (NSAID) chronic treatment on cognitive status in non-demented elderly patients. Sixty-six patients (25.6%) were considered chronic NSAID users. Patients chronically assuming NSAADs showed a significantly higher MMSE score than non-users (26.9+/-2.1 vs 25.7+/-2.5, P<0.0005 ). After controlling for potential confounders in a multivariate model, chronic NSAID use remained independently associated with MMSE score. The results support a positive association between chronic NSAID use and cognitive function in non-demented elderly patients. Randomized controlled trials will be needed to definitively prove this beneficial effect.

Factors related to length of stay in a geriatric evaluation and rehabilitation unit.

Factors related to length of stay were examined in 295 elderly patients (mean age = 79.0 +/- 7.3, range 65-94; males = 75, females = 220), consecutively admitted to a Geriatric Evaluation and Rehabilitation Unit (GERU, P. Richiedei Hospital, Gussago, Brescia, Italy) over a twelve-month period (November 1, 1993-October 31, 1994). Mini Mental State Examination (MMSE), Geriatric Depression Scale (GDS), Basic Activities of Daily Living (BADL), Instrumental Activities of Daily Living (IADL), Tinetti Scale, Prognostic Nutritional Index (PNI), number of diseases and number of administered drugs were evaluated. An index of Disease Severity (IDS) was utilized to estimate the level of comorbidity severity. Three comorbidity classes were thus defined: I) patients with no disease of relevant severity; II) patients with only one disease of relevant severity accompanied by clinically significant comorbidity; and III) patients with two or more relevant diseases. The variables associated with the length of stay proved to be classes of comorbidity, MMSE, dependence in BADL and IADL, Tinetti scale, and PNI. The association of longer length of stay with greater comorbidity was enhanced by impairment in gait and balance (Tinetti < 18) and malnutrition (PNI > 45). These data suggest that the length of stay in hospital is related to comorbidity in patients with conditions of physical and biomedical frailty.

Mood improvement in elderly women after in-hospital physical rehabilitation.

OBJECTIVE: To evaluate the relationship between change in depressive symptoms and in-hospital physical rehabilitation in elderly women. DESIGN: Longitudinal study. SETTING: Hospital facility (geriatric evaluation and rehabilitation unit). PATIENTS: One hundred twenty-three elderly inpatient women (mean age: 78.4+/-6.9 years, range 60 to 93) with good cognitive status (Mini Mental State Examination: 23.1+/-5.1) consecutively admitted over a 7-month period. INTERVENTION: Physical therapy tailored to individual needs (five sessions a week of 30 to 45 minutes each). MAIN OUTCOMES MEASURES: On admission: cognition (MMSE), depressive symptoms (Geriatric Depression Scale [GDS]), functional status (basic and instrumental activities of daily living [BADL, IADL], Tinetti scale), and somatic health. On discharge: depressive symptoms and gait and balance performances (Tinetti scale). RESULTS: Seventy-five patients (61%) did not show changes on Tinetti scale over the hospitalization period and 48(39%) had a change of 3 or more points. Nonresponders had no change of GDS over the hospitalization period for all levels of physical disability on admission, whereas responders had relevant improvement of depressive symptoms when markedly disabled on admission, and progressively smaller improvements of depressive symptoms with increasing function on admission. CONCLUSIONS: The study provides evidence that mood status changes synchronically with disability.

Assessment of functional ability with the bed rise difficulty scale in a group of elderly patients.

The aim of the study (part of the Progetto Longitudinale Gussago) was to evaluate the variables related to the difficulty in rising from a bed in 2 groups of elderly patients: nursing home residents, and patients admitted to a geriatric evaluation and management unit. Functional ability was tested through the bed rise difficulty scale (BRD). The version used in this study considered only those 7 items (out of 12) found to be of value. Only those patients who were able to rise from bed without help were selected in order to achieve the aim of the study (33 males, 113 females; mean age 79.6 +/- 7.3 years). Although the 146 patients assessed were considered as having a good functional level (Tinetti score 18.8 +/- 6.9, ADL Katz score 1.6 +/- 1.4), most of them had high scores on the BRD scale, indicating the ability of this scale to detect early, mild disability. The total score of the BRD scale was significantly related to the ADL Katz (r = 0.29, p = 0.000), Tinetti scale (r = -0.39, p = 0.000) and physical performance test (PTT; r = -0.47, p = 0.000). Similar results were obtained for the correlation between BRD time and ADL Katz (r = 0.033, p = 0.000), Tinetti scale (r = -0.30, p = 0.000) and PPT-(r = -0.46, p = 0.000). In a logistic regression analysis the items of the PPT scale considering upper extremity function and Tinetti balance score were significantly associated with the total bed rise time and score.

Food intake and mortality in the frail elderly.

BACKGROUND: Adequate qualitative and quantitative food intake is a major determinant of health. However, nutritional requirements in the elderly are unknown, and even more so in the frail elderly. The aim of the study was to evaluate the influence of energy and macro-nutrients on health in the frail nursing home elderly. METHODS: Food intake of 72 not severely diseased elderly patients was assessed with direct weighing method. Outcome measure was survival over 28-month follow-up period. Confounders of the association of food intake with survival were: age, gender, body-mass index, daily function, somatic health, anergy, and nutritional status. Crude association of food intake with survival was assessed with Kaplan-Meyer method, and adjusted association with multiple Cox regression models. RESULTS: Patients of the study had good average food intake. Mortality rate was relatively low (.20 per year). Low levels of energy, protein, lipid, and carbohydrate intake were negatively associated with survival even after adjustment for confounders. When compared to high intake, adjusted relative risks for mortality of low intake were 4.74, 3.75, 4.71, and 2.04, respectively. Medium levels of energy, protein, and lipid, but not carbohydrate, intake yielded intermediate mortality risk. CONCLUSIONS: Food intake is a strong predictor of survival even in moderately diseased elderly patients, suggesting possible low-cost interventions.