Culturing and Molecular Approaches for Identifying Microbiota Taxa Impacting Children's Obesogenic Phenotypes Related to Xenobiotic Dietary Exposure.

Integrated data from molecular and improved culturomics studies might offer holistic insights on gut microbiome dysbiosis triggered by xenobiotics, such as obesity and metabolic disorders. Bisphenol A (BPA), a dietary xenobiotic obesogen, was chosen for a directed culturing approach using microbiota specimens from 46 children with obesity and normal-weight profiles. In parallel, a complementary molecular analysis was carried out to estimate the BPA metabolising capacities. Firstly, catalogues of 237 BPA directed-cultured microorganisms were isolated using five selected media and several BPA treatments and conditions. Taxa from Firmicutes, Proteobacteria, and Actinobacteria were the most abundant in normal-weight and overweight/obese children, with species belonging to the genera Enterococcus, Escherichia, Staphylococcus, Bacillus, and Clostridium. Secondly, the representative isolated taxa from normal-weight vs. overweight/obese were grouped as BPA biodegrader, tolerant, or resistant bacteria, according to the presence of genes encoding BPA enzymes in their whole genome sequences. Remarkably, the presence of sporobiota and concretely Bacillus spp. showed the higher BPA biodegradation potential in overweight/obese group compared to normal-weight, which could drive a relevant role in obesity and metabolic dysbiosis triggered by these xenobiotics.

Antimicrobial Effects of Potential Probiotics of Bacillus spp. Isolated from Human Microbiota: In Vitro and In Silico Methods.

The variable taxa components of human gut microbiota seem to have an enormous biotechnological potential that is not yet well explored. To investigate the usefulness and applications of its biocompounds and/or bioactive substances would have a dual impact, allowing us to better understand the ecology of these microbiota consortia and to obtain resources for extended uses. Our research team has obtained a catalogue of isolated and typified strains from microbiota showing resistance to dietary contaminants and obesogens. Special attention was paid to cultivable Bacillus species as potential next-generation probiotics (NGP) together with their antimicrobial production and ecological impacts. The objective of the present work focused on bioinformatic genome data mining and phenotypic analyses for antimicrobial production. In silico methods were applied over the phylogenetically closest type strain genomes of the microbiota Bacillus spp. isolates and standardized antimicrobial production procedures were used. The main results showed partial and complete gene identification and presence of polyketide (PK) clusters on the whole genome sequences (WGS) analysed. Moreover, specific antimicrobial effects against B. cereus, B. circulans, Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Serratia marcescens, Klebsiella spp., Pseudomonas spp., and Salmonella spp. confirmed their capacity of antimicrobial production. In conclusion, Bacillus strains isolated from human gut microbiota and taxonomic group, resistant to Bisphenols as xenobiotics type endocrine disruptors, showed parallel PKS biosynthesis and a phenotypic antimicrobial effect. This could modulate the composition of human gut microbiota and therefore its functionalities, becoming a predominant group when high contaminant exposure conditions are present.

Memory and Executive Dysfunction Predict Complex Activities of Daily Living Impairment in Amnestic Multi-Domain Mild Cognitive Impairment.

BACKGROUND: Specific cognitive alterations could be one of the predictors that lead to the complex activities of daily living (CADL) impairment in mild cognitive impairment (MCI) and, hence, help to explain the continuum between MCI and dementia. OBJECTIVE: We aimed to reevaluate the existing uncertainty regarding the impact of memory and executive functions on CADL in patients with MCI. METHODS: Caregivers of 161 patients with amnestic multi-domain MCI and of 150 patients with incipient Alzheimer's disease as well as 100 age-, sex-, and education-matched controls, completed the Interview for Deterioration in Daily Living Activities in Dementia, a suitable instrument for the description and discrimination of CADL. In addition, all patients and controls were assessed with a neuropsychological battery to measure explicit memory and executive functions performance. RESULTS: Multiple regression analyses showed that in the group of patients with amnestic multi-domain MCI, 67.4% of the variability of the CADL impairment was explained by worse performance on executive functions tests (p < 0.0001) and 41.8% by different explicit memory components impairment (p < 0.0001). Further, in patients with incipient AD, 44.0% of the variability of CADL impairment was explained by worse performance on executive functions tests (p < 0.0001) and 39.9% by different explicit memory components worsening (p < 0.0001). CONCLUSIONS: Memory and executive functions alterations impact similarly on the CADL in both amnestic multi-domain MCI and incipient Alzheimer's disease. Given the continuum that exists between both conditions, we conclude that CADL impairment may be an important early step in the evolution towards Alzheimer's disease from amnestic multi-domain MCI.

Neurological Disorders in Central Spain, Second Survey: Feasibility Pilot Observational Study.

BACKGROUND: The Neurological Disorders in Central Spain, second survey (NEDICES-2) is a population-based, closed-cohort study that will include over 8000 subjects aged ≥55 years. It will also include a biobank. OBJECTIVE: The objective of this study was to evaluate all major aspects of the NEDICES-2 (methods, database, screening instruments, and questionnaires, as well as interexpert rating of the neurological diagnoses) in each one of the planned areas (all of them in central Spain) and to test the possibility of obtaining biological samples from each participant. METHODS: A selection of patients and participants of the planned NEDICES-2 underwent face-to-face interviews including a comprehensive questionnaire on demographics, current medications, medical conditions, and lifestyle habits. Biological samples (blood, saliva, urine, and hair) were also obtained. Furthermore, every participant was examined by a neurologist. RESULTS: In this pilot study, 567 study participants were enrolled (196 from hospitals and 371 from primary care physician lists). Of these 567, 310 completed all study procedures (questionnaires and the neurological evaluation). The study was time-consuming for several primary care physicians. Hence, a few primary care physicians from some areas refused to participate, which led to a reconfiguration of study areas. In addition, the central biobank needed to be supplemented by the biobanks of local Spanish National Health System hospitals. CONCLUSIONS: Population-based epidemiological surveys, such as the NEDICES-2, require a pilot study to evaluate the feasibility of all aspects of a future field study (population selection, methods and instruments to be used, neurological diagnosis agreement, and data collection).

Harmonization of the quantitative determination of volatile fatty acids profile in aqueous matrix samples by direct injection using gas chromatography and high-performance liquid chromatography techniques: Multi-laboratory validation study.

The performance parameters of volatile fatty acids (VFAs) measurements were assessed for the first time by a multi-laboratory validation study among 13 laboratories. Two chromatographic techniques (GC and HPLC) and two quantification methods such as external and internal standard (ESTD/ISTD) were combined in three different methodologies GC/ESTD, HPLC/ESTD and GC/ISTD. Linearity evaluation of the calibration functions in a wide concentration range (10-1000mg/L) was carried out using different statistical parameters for the goodness of fit. Both chromatographic techniques were considered similarly accurate. The use of GC/ISTD, despite showing similar analytical performance to the other methodologies, can be considered useful for the harmonization of VFAs analytical methodology taking into account the normalization of slope values used for the calculation of VFAs concentrations. Acceptance criteria for VFAs performance parameters of the multi-laboratory validation study should be established as follows: (1) instrument precision (RSDINST≤1.5%); (2) linearity (R(2)≥0.998; RSDSENSITIVITY≤4%; REMAX≤8%; REAVER≤ 3%); (3) precision (RSD≤1.5%); (4) trueness (recovery of 97-103%); (5) LOD (≤3mg/L); and (6) LOQ (10mg/L).

Methods and design of the baseline survey of the neurological disorders in Salamanca (NEDISA) cohort: a population-based study in Central-Western Spain.

BACKGROUND: To describe the design of the baseline assessment of an epidemiological study of elderly persons living in Salamanca, central-western Spain: the Neurological Diseases in Salamanca (NEDISA) study. We assessed the epidemiology of stroke, cognitive disorders, essential tremor (ET), Parkinson's disease (PD) and restless legs syndrome. METHODS: In phase 1 (February 1 to May 31, 2007), 4 neurologists and 2 trained general physicians examined and performed phlebotomy on all participants. In phase 2 (June 1, 2007, to June 1, 2008), the participants were reexamined and had a complete neuropsychological assessment. Neuroimaging was performed in participants with cognitive disorders, ET and PD. RESULTS: The registered study population consisted of 1,077 individuals, but 45 people were ineligible (address change, refusals or death), leaving a final sample of 1,032 (95.8%). The main demographic data on the 1,032 participants (408 men, 624 women) are provided. CONCLUSIONS: Most of the registered study population was enrolled, and this may have been due to the close relationship between NEDISA researchers and the general physicians in the area of study. The NEDISA study will likely improve our knowledge of prevalence rates of the neurological diseases chosen for study as well as the set of risk factors that predispose individuals in Spain to these disorders.

Nonverbal memory tasks in early differential diagnosis of Alzheimer's disease and unipolar depression.

We examined whether there are selective deficits in early-stage Alzheimer's disease (AD; n = 27) and in unipolar depression (UD; n = 17) patients on recall and recognition of spatial and visual components of nonverbal memory (NVM) and whether the two groups can be differentiated based on their performance on such tasks. We also investigated which NVM measures had the best discrimination power. We tested spatial, visuospatial, and visuoconstructive abilities in AD and UD patients. AD patients' scores on NVM tasks were significantly lower than those of healthy subjects (HS; n = 30) and consistently lower than those of the UD group. Z-scores suggested that AD patients suffered from a generalized impairment. Clear differences between AD and UD patients were found on abstract design tasks.

Does the combination of the MMSE and clock drawing test (mini-clock) improve the detection of mild Alzheimer's disease and mild cognitive impairment?

There is currently a need to develop tools to identify patients with mild AD and mild cognitive impairment (MCI). We determined the validity and reliability of a brief, easily administered cognitive screening battery consisting of fusion of two well-known brief tests (Mini-Mental Status Examination [MMSE] and Clock Drawing Test [CDT]) (Mini-clock) to differentiate between patients with mild AD, MCI, and healthy control subjects. 66 consecutive patients with mild AD, 21 with MCI, and 66 healthy controls seen in a memory clinic setting were compared. Receiver operating characteristic (ROC) curve analysis was used to calculate the cut-off value permitting discrimination between mild AD, MCI, and healthy control subjects. Interrater and test-retest reliability were also assessed. Mean cognitive scores for patients with AD, MCI, and control subjects on all two individual tests were significantly different (for each, p < 0.001). The mean area under the ROC curve for Mini-clock was higher than that obtained with MMSE or CDT in differentiating mild AD from controls (0.973 vs. 0.952 and 0.881, respectively) and MCI from controls (0.855 vs. 0.821 and 0.779, respectively). Test-retest reliability for the Mini-clock was 0.99, meanwhile interrater reliability was 0.87. The mean time to complete the test for all subjects was 8 min and 50 s. The Mini-clock is highly sensitive and specific in the detection of mild AD and reasonably accurate when attempting to separate MCI from health controls. It has a high interrater and test-retest reliability, can be quickly administered, and does not require major training.

Improvement pattern in the clock drawing test in early Alzheimer's disease.

OBJECTIVE: The aim of this paper was to compare the performance of a group of patients with early Alzheimer's disease (EAD) against a control group of healthy control (HC) subjects in the Clock Drawing Test (CDT), i.e. verbal command versus copying of a clock model presented to the subject. PATIENTS AND METHODS: The authors have studied 140 subjects; 70 patients with probable EAD, with a mean age of 76.4 +/- 7.64 years and a clinical dementia rating stage 1 (mild dementia), and 70 HC with a mean age of 75.16 +/- 6.34 years. RESULTS: Patients in the EAD group obtained significantly higher scores on the copy command mode than on the verbal command mode (Z = -7.129, p < 0.001)--improvement pattern of the CDT--whereas no statistically significant differences were found in the HC group (Z = -2.001, p < 0.080). Within the group of EAD patients, we have noticed that there is a correlation between the copy command mode and the visual-constructive functions of the Cambridge Cognitive Examination (CAMCOG) (r = 0.607, p < 0.01), while the memory functions of the CAMCOG correlate with the verbal command mode (r = 0.704, p < 0.01). CONCLUSIONS: In our study, the EAD patients show an improvement pattern in the execution of the CDT copy command in comparison with the execution of the CDT verbal command, which we did not observe in the HC group. Such results might be associated with a greater deterioration of the memory functions when compared with the visual-constructive ones in the patients with EAD.