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1.  A two-year follow-up of cognitive deficits and brain perfusion in Mild Cognitive Impairment and mild Alzheimer’s disease 
Background
The 15-Objects Test (15-OT) provides useful gradation of visuoperceptual impairment from normal aging through Alzheimer’s disease (AD) and correlates with temporo-parietal perfusion.
Objectives
To analyse progression of 15-OT performance in Mild Cognitive Impairment (MCI) and AD, and its correlates with cognition and Single Photon Emission Computerized Tomography (SPECT). Further, to examine neuropsychological and SPECT differences between the MCI patients who developed AD and those who didn’t.
Methods
From the initial 126 participants (42/group), 38AD, 39MCI and 38 elderly controls (EC) were reassessed (SPECT: 35AD, 33MCI, 35EC) after two years. The progression of cognitive and SPECT scores during this period was compared between groups, and baseline data between converters and non-converters. SPECT data were analysed by SPM5.
Results
The 15-OT was the only measure of progression that differed between the three groups; worsening scores on 15-OT were associated with worsening in verbal and visual retention, and decreased perfusion on left postsubicular area. In the MCI patients cerebral perfusion fell over the two years in medial-posterior cingulate and fronto-temporo-parietal regions; AD showed extensive changes involving almost all cerebral regions. No SPECT changes were detected in controls. At baseline, the MCI patients who developed AD differed from non-converters in verbal recognition memory, but not in SPECT perfusion.
Conclusion
SPECT and 15-OT appear to provide a potential measure to differenciate between progression of normal aging, MCI and AD. Worsening on 15-OT was related to decreased perfusion in postsubicular area; but further longitudinal studies are needed to determine the contribution of 15-OT as a predictor of AD from MCI.
doi:10.3233/JAD-2012-111850
PMCID: PMC3589736  PMID: 22406443
Visuoperception; The 15-Objects test; cerebral perfusion; brain SPECT; two-year follow-up; prospective; longitudinal; Mild Cognitive Impairment; Alzheimer’s disease
2.  Brain perfusion correlates of visuoperceptual deficits in Mild Cognitive Impairment and mild Alzheimer’s disease 
Journal of Alzheimer's Disease  2010;21(2):557-567.
Background
Visuoperceptual processing is impaired early in the clinical course of Alzheimer’s disease (AD). The 15-Objects Test (15-OT) detects such subtle performance deficits in Mild Cognitive Impairment (MCI) and mild AD. Reduced brain perfusion in the temporal, parietal and prefrontal regions have been found in early AD and MCI patients.
Objectives
To confirm the role of the 15-OT in the diagnosis of MCI and AD, and to investigate the brain perfusion correlates of visuoperceptual dysfunction (15-OT) in subjects with MCI, AD and normal aging.
Methods
Forty-two AD, 42 MCI and 42 healthy elderly control (EC) subjects underwent a brain Single Photon Emission Tomography (SPECT) and separately completed the 15-OT. An analysis of variance compared 15-OT scores between groups. SPM5 was used to analyse the SPECT data.
Results
15-OT performace was impaired in the MCI and AD patients. In terms of the SPECT scans, AD patients showed reduced perfusion in temporal-parietal regions, while the MCI subjects had decreased perfusion in the middle and posterior cingulate. When MCI and AD groups were compared, a significant brain perfusion reduction was found in temporo-parietal regions. In the whole sample, 15-OT performance was significantly correlated with the clinical dementia rating scores, and with the perfusion in the bilateral posterior cingulate and the right temporal pole, with no significant correlation in each separate group.
Conclusion
Our findings suggest that the 15-OT performance provides a useful gradation of impairment from normal aging to AD, and it seems to be related to perfusion in the bilateral posterior cingulate and the right temporal pole.
doi:10.3233/JAD-2010-091069
PMCID: PMC3306804  PMID: 20555146
Visuoperception; cerebral perfusion; brain SPECT; Mild Cognitive Impairment; Alzheimer’s disease
3.  Comparative Skull Analysis Suggests Species-Specific Captivity-Related Malformation in Lions (Panthera leo) 
PLoS ONE  2014;9(4):e94527.
Lion (Panthera leo) populations have dramatically decreased worldwide with a surviving population estimated at 32,000 across the African savannah. Lions have been kept in captivity for centuries and, although they reproduce well, high rates of stillbirths as well as morbidity and mortality of neonate and young lions are reported. Many of these cases are associated with bone malformations, including foramen magnum (FM) stenosis and thickened tentorium cerebelli. The precise causes of these malformations and whether they are unique to captive lions remain unclear. To test whether captivity is associated with FM stenosis, we evaluated 575 lion skulls of wild (N = 512) and captive (N = 63) origin. Tiger skulls (N = 276; 56 captive, 220 wild) were measured for comparison. While no differences were found between males and females or between subadults and adults in FM height (FMH), FMH of captive lions (17.36±3.20 mm) was significantly smaller and with greater variability when compared to that in wild lions (19.77±2.11 mm). There was no difference between wild (18.47±1.26 mm) and captive (18.56±1.64 mm) tigers in FMH. Birth origin (wild vs. captive) as a factor for FMH remained significant in lions even after controlling for age and sex. Whereas only 20/473 wild lions (4.2%) had FMH equal to or smaller than the 5th percentile of the wild population (16.60 mm), this was evident in 40.4% (23/57) of captive lion skulls. Similar comparison for tigers found no differences between the captive and wild populations. Lions with FMH equal to or smaller than the 5th percentile had wider skulls with smaller cranial volume. Cranial volume remained smaller in both male and female captive lions when controlled for skull size. These findings suggest species- and captivity-related predisposition for the pathology in lions.
doi:10.1371/journal.pone.0094527
PMCID: PMC3981823  PMID: 24718586
4.  The Influence of Social Structure, Habitat, and Host Traits on the Transmission of Escherichia coli in Wild Elephants 
PLoS ONE  2014;9(4):e93408.
Social structure is proposed to influence the transmission of both directly and environmentally transmitted infectious agents. However in natural populations, many other factors also influence transmission, including variation in individual susceptibility and aspects of the environment that promote or inhibit exposure to infection. We used a population genetic approach to investigate the effects of social structure, environment, and host traits on the transmission of Escherichia coli infecting two populations of wild elephants: one in Amboseli National Park and another in Samburu National Reserve, Kenya. If E. coli transmission is strongly influenced by elephant social structure, E. coli infecting elephants from the same social group should be genetically more similar than E. coli sampled from members of different social groups. However, we found no support for this prediction. Instead, E. coli was panmictic across social groups, and transmission patterns were largely dominated by habitat and host traits. For instance, habitat overlap between elephant social groups predicted E. coli genetic similarity, but only in the relatively drier habitat of Samburu, and not in Amboseli, where the habitat contains large, permanent swamps. In terms of host traits, adult males were infected with more diverse haplotypes, and males were slightly more likely to harbor strains with higher pathogenic potential, as compared to adult females. In addition, elephants from similar birth cohorts were infected with genetically more similar E. coli than elephants more disparate in age. This age-structured transmission may be driven by temporal shifts in genetic structure of E. coli in the environment and the effects of age on bacterial colonization. Together, our results support the idea that, in elephants, social structure often will not exhibit strong effects on the transmission of generalist, fecal-oral transmitted bacteria. We discuss our results in the context of social, environmental, and host-related factors that influence transmission patterns.
doi:10.1371/journal.pone.0093408
PMCID: PMC3976290  PMID: 24705319
5.  Globally Expanding Carbapenemase Finally Appears in Spain: Nosocomial Outbreak of Acinetobacter baumannii Producing Plasmid-Encoded OXA-23 in Barcelona, Spain 
Antimicrobial Agents and Chemotherapy  2013;57(10):5155-5157.
Resistance of Acinetobacter baumannii clinical isolates to carbapenems is on the rise worldwide mainly in association with the production of OXA-23. Until recently, however, OXA-23 was absent in Spain. In this work, we report the molecular characterization of a hospital outbreak of OXA-23-producing A. baumannii in Barcelona caused by a multidrug-resistant (MDR) clone belonging to international clone IC-II/sequence type ST85 between October 2010 and May 2011. blaOXA-23 was carried in a plasmid of 90 kb and located within the composite transposon Tn2006.
doi:10.1128/AAC.01486-13
PMCID: PMC3811394  PMID: 23877694
6.  Is the Physician’s Behavior in Dyslipidemia Diagnosis in Accordance with Guidelines? Cross-Sectional Escarval Study 
PLoS ONE  2014;9(3):e91567.
Background
Clinical inertia has been defined as mistakes by the physician in starting or intensifying treatment when indicated. Inertia, therefore, can affect other stages in the healthcare process, like diagnosis. The diagnosis of dyslipidemia requires ≥2 high lipid values, but inappropriate behavior in the diagnosis of dyslipidemia has only previously been analyzed using just total cholesterol (TC).
Objectives
To determine clinical inertia in the dyslipidemia diagnosis using both TC and high-density lipoprotein cholesterol (HDL-c) and its associated factors.
Design
Cross-sectional.
Setting
All health center visits in the second half of 2010 in the Valencian Community (Spain).
Patients
11,386 nondyslipidemic individuals aged ≥20 years with ≥2 lipid determinations.
Measurement Variables
Gender, atrial fibrillation, hypertension, diabetes, cardiovascular disease, age, and ESCARVAL training course. Lipid groups: normal (TC<5.17 mmol/L and normal HDL-c [≥1.03 mmol/L in men and ≥1.29 mmol/L in women], TC inertia (TC≥5.17 mmol/L and normal HDL-c), HDL-c inertia (TC<5.17 mmol/L and low HDL-c), and combined inertia (TC≥5.17 mmol/L and low HDL-c).
Results
TC inertia: 38.0% (95% CI: 37.2–38.9%); HDL-c inertia: 17.7% (95% CI: 17.0–18.4%); and combined inertia: 9.6% (95% CI: 9.1–10.2%). The profile associated with TC inertia was: female, no cardiovascular risk factors, no cardiovascular disease, middle or advanced age; for HDL-c inertia: female, cardiovascular risk factors and cardiovascular disease; and for combined inertia: female, hypertension and middle age.
Limitations
Cross-sectional study, under-reporting, no analysis of some cardiovascular risk factors or other lipid parameters.
Conclusions
A more proactive attitude should be adopted, focusing on the full diagnosis of dyslipidemia in clinical practice. Special emphasis should be placed on patients with low HDL-c levels and an increased cardiovascular risk.
doi:10.1371/journal.pone.0091567
PMCID: PMC3953440  PMID: 24626597
7.  Understanding Light Harvesting in Radial Junction Amorphous Silicon Thin Film Solar Cells 
Scientific Reports  2014;4:4357.
The radial junction (RJ) architecture has proven beneficial for the design of a new generation of high performance thin film photovoltaics. We herein carry out a comprehensive modeling of the light in-coupling, propagation and absorption profile within RJ thin film cells based on an accurate set of material properties extracted from spectroscopic ellipsometry measurements. This has enabled us to understand and evaluate the impact of varying several key parameters on the light harvesting in radially formed thin film solar cells. We found that the resonance mode absorption and antenna-like light in-coupling behavior in the RJ cell cavity can lead to a unique absorption distribution in the absorber that is very different from the situation expected in a planar thin film cell, and that has to be taken into account in the design of high performance RJ thin film solar cells. When compared to the experimental EQE response of real RJ solar cells, this modeling also provides an insightful and powerful tool to resolve the wavelength-dependent contributions arising from individual RJ units and/or from strong light trapping due to the presence of the RJ cell array.
doi:10.1038/srep04357
PMCID: PMC3950579  PMID: 24619197
8.  Minimally Invasive Robotic Kidney Transplantation for Obese Patients Previously Denied Access to Transplantation 
Obese patients with end-stage renal disease (ESRD) are often excluded from kidney transplantation due to concerns about surgical site infections. To reduce infections, we developed a robotic kidney transplantation method for obese recipients. From June 2009-December 2011, a prospective cohort of 39 obese patients underwent robotic kidney transplantation at a single center. The outcomes of patients with at least six months of follow-up (n=28) were compared to a frequency-matched retrospective cohort of obese patients who underwent open kidney transplantation from 2004-2009 (n=28). The 28 robotic patients were predominately African-American (46.4%) or Hispanic (35.7%), with a mean age of 47.9±10.7 years, similar to the control group. BMI in the robotic group was 42.6±7.8 kg/m2 compared to 38.1±5.4 kg/m2 in the control group (p=0.02). There were no surgical site infections in the robotic group (0/28), while 28.6% (8/28) in the control group developed an infection (p=0.004). Six-month creatinine (1.5±0.4 vs.1.6±0.6 mg/dL; p=0.47), and patient and graft survival (100%) were comparable between the two groups. Outcomes following robotic surgery compared favorably to conventional transplantation. Robotic surgery may therefore enable obese patients with ESRD to access kidney transplantation and may thereby reduce health disparities in groups with a high prevalence of obesity and ESRD.
doi:10.1111/ajt.12078
PMCID: PMC3647345  PMID: 23437881
Robotic Surgery; Kidney Transplant; Obesity; Health Disparities
9.  Anal gas evacuation and colonic microbiota in patients with flatulence: effect of diet 
Gut  2013;63(3):401-408.
Objective
To characterise the influence of diet on abdominal symptoms, anal gas evacuation, intestinal gas distribution and colonic microbiota in patients complaining of flatulence.
Design
Patients complaining of flatulence (n=30) and healthy subjects (n=20) were instructed to follow their usual diet for 3 days (basal phase) and to consume a high-flatulogenic diet for another 3 days (challenge phase).
Results
During basal phase, patients recorded more abdominal symptoms than healthy subjects in daily questionnaires (5.8±0.3 vs 0.4±0.2 mean discomfort/pain score, respectively; p=<0.0001) and more gas evacuations by an event marker (21.9±2.8 vs 7.4±1.0 daytime evacuations, respectively; p=0.0001), without differences in the volume of gas evacuated after a standard meal (262±22 and 265±25 mL, respectively). On flatulogenic diet, both groups recorded more abdominal symptoms (7.9±0.3 and 2.8±0.4 discomfort/pain, respectively), number of gas evacuations (44.4±5.3 and 21.7±2.9 daytime evacuations, respectively) and had more gas production (656±52 and 673±78 mL, respectively; p<0.05 vs basal diet for all). When challenged with flatulogenic diet, patients’ microbiota developed instability in composition, exhibiting variations in the main phyla and reduction of microbial diversity, whereas healthy subjects’ microbiota were stable. Taxa from Bacteroides fragilis or Bilophila wadsworthia correlated with number of gas evacuations or volume of gas evacuated, respectively.
Conclusions
Patients complaining of flatulence have a poor tolerance of intestinal gas, which is associated with instability of the microbial ecosystem.
doi:10.1136/gutjnl-2012-303013
PMCID: PMC3933177  PMID: 23766444
Colonic Bacteria; Colonic Fermentation; Colonic Microflora; Functional Bowel Disorder; Visceral Sensitivity
10.  Telemedicine-Based Approach for Obstructive Sleep Apnea Management: Building Evidence 
Background
Telemedicine seems to offer reliable solutions to health care challenges, but significant contradictory results were recently found. Therefore, it is crucial to carefully select outcomes and target patients who may take advantage of this technology. Continuous positive airway pressure (CPAP) therapy compliance is essential to treat patients with obstructive sleep apnea (OSA). We believe that OSA patients could benefit greatly from a telemedicine approach for CPAP therapy management.
Objective
The objective of our study was to evaluate the application of a telemedicine-based approach in the CPAP therapy management, focusing on patients’ CPAP follow-up and training.
Methods
We performed two studies. First, (study 1) we enrolled 50 consecutive OSA patients who came to our sleep center for the CPAP follow-up visit. Patients performed a teleconsultation with a physician, and once finalized, they were asked to answer anonymously to a questionnaire regarding their opinion about the teleconsultation. In a second randomized controlled trial (RCT) (study 2), we included 40 OSA patients scheduled for CPAP training. There were 20 that received the usual face-to-face training and 20 that received the training via videoconference. After the session, they were blindly evaluated on what they learned about OSA and mask placement.
Results
More than 95% (49/50) of the interviewed patients were satisfied with the teleconsultation, and 66% (33/50) of them answered that the teleconsultation could replace 50%-100% of their CPAP follow-up visits. Regarding the RCT, patients who received the CPAP training via videoconference demonstrated the same knowledge about OSA and CPAP therapy as the face-to-face group (mean 93.6% of correct answers vs mean 92.1%; P=.935). Performance on practical skills (mask and headgear placement, leaks avoidance) was also similar between the two groups.
Conclusions
OSA patients gave a positive feedback about the use of teleconsultation for CPAP follow-up, and the CPAP training based on a telemedicine approach proved to be as effective as face-to-face training. These results support the use of this telemedicine-based approach as a valuable strategy for patients’ CPAP training and clinical follow-up.
doi:10.2196/ijmr.3060
PMCID: PMC3961625  PMID: 24554392
telemedicine; sleep apnea; CPAP therapy; teleconsultation
11.  Modifying the Victor® Easy Set® Rat Trap to Improve the Animal Welfare of Stoats and Ship Rats Trapped in New Zealand 
PLoS ONE  2014;9(2):e86760.
Stoats (Mustela erminea) and ship rats (Rattus rattus) in New Zealand are targeted by trapping to mitigate their predation on native wildlife. Internationally recognized guidelines for assessing the effectiveness and welfare performance of kill traps are supported by New Zealand legislation under the Animal Welfare Act 1999. The Victor® Easy Set® rat trap was tested and passed a similar standard for killing short-tailed weasels in Canada but failed for stoats when tested in New Zealand in 2002 (short-tailed weasels and stoats are the same species). We tested a modified version of the trap in 2011–12, modified by changing the treadle trigger to a pull trigger and adding a plastic shroud to direct and align approach by animals to the front of the trap. These traps, in vertical and horizontal sets, were tested with both stoats and ship rats. During each test the trap had to render 10 of 10 animals irreversibly unconscious within 3 minutes to meet approval requirements. The modified trap passed with both species in both trap sets. All stoats were struck across the cranium whereas rats were struck either on the cranium or neck. We recommend this trap design for use by community conservation groups for targeting stoats and ship rats in New Zealand.
doi:10.1371/journal.pone.0086760
PMCID: PMC3914800  PMID: 24505264
12.  A Metapopulation Approach to African Lion (Panthera leo) Conservation 
PLoS ONE  2014;9(2):e88081.
Due to anthropogenic pressures, African lion (Panthera leo) populations in Kenya and Tanzania are increasingly limited to fragmented populations. Lions living on isolated habitat patches exist in a matrix of less-preferred habitat. A framework of habitat patches within a less-suitable matrix describes a metapopulation. Metapopulation analysis can provide insight into the dynamics of each population patch in reference to the system as a whole, and these analyses often guide conservation planning. We present the first metapopulation analysis of African lions. We use a spatially-realistic model to investigate how sex-biased dispersal abilities of lions affect patch occupancy and also examine whether human densities surrounding the remaining lion populations affect the metapopulation as a whole. Our results indicate that male lion dispersal ability strongly contributes to population connectivity while the lesser dispersal ability of females could be a limiting factor. When populations go extinct, recolonization will not occur if distances between patches exceed female dispersal ability or if females are not able to survive moving across the matrix. This has profound implications for the overall metapopulation; the female models showed an intrinsic extinction rate from five-fold to a hundred-fold higher than the male models. Patch isolation is a consideration for even the largest lion populations. As lion populations continue to decline and with local extinctions occurring, female dispersal ability and the proximity to the nearest lion population are serious considerations for the recolonization of individual populations and for broader conservation efforts.
doi:10.1371/journal.pone.0088081
PMCID: PMC3914926  PMID: 24505385
13.  Nonviral-Mediated Hepatic Expression of IGF-I Increases Treg Levels and Suppresses Autoimmune Diabetes in Mice 
Diabetes  2013;62(2):551-560.
In type 1 diabetes, loss of tolerance to β-cell antigens results in T-cell–dependent autoimmune destruction of β cells. The abrogation of autoreactive T-cell responses is a prerequisite to achieve long-lasting correction of the disease. The liver has unique immunomodulatory properties and hepatic gene transfer results in tolerance induction and suppression of autoimmune diseases, in part by regulatory T-cell (Treg) activation. Hence, the liver could be manipulated to treat or prevent diabetes onset through expression of key genes. IGF-I may be an immunomodulatory candidate because it prevents autoimmune diabetes when expressed in β cells or subcutaneously injected. Here, we demonstrate that transient, plasmid-derived IGF-I expression in mouse liver suppressed autoimmune diabetes progression. Suppression was associated with decreased islet inflammation and β-cell apoptosis, increased β-cell replication, and normalized β-cell mass. Permanent protection depended on exogenous IGF-I expression in liver nonparenchymal cells and was associated with increased percentage of intrapancreatic Tregs. Importantly, Treg depletion completely abolished IGF-I-mediated protection confirming the therapeutic potential of these cells in autoimmune diabetes. This study demonstrates that a nonviral gene therapy combining the immunological properties of the liver and IGF-I could be beneficial in the treatment of the disease.
doi:10.2337/db11-1776
PMCID: PMC3554392  PMID: 23099863
14.  Follow-up of loci from the International Genomics of Alzheimer's Disease Project identifies TRIP4 as a novel susceptibility gene 
Translational Psychiatry  2014;4(2):e358-.
To follow-up loci discovered by the International Genomics of Alzheimer's Disease Project, we attempted independent replication of 19 single nucleotide polymorphisms (SNPs) in a large Spanish sample (Fundació ACE data set; 1808 patients and 2564 controls). Our results corroborate association with four SNPs located in the genes INPP5D, MEF2C, ZCWPW1 and FERMT2, respectively. Of these, ZCWPW1 was the only SNP to withstand correction for multiple testing (P=0.000655). Furthermore, we identify TRIP4 (rs74615166) as a novel genome-wide significant locus for Alzheimer's disease risk (odds ratio=1.31; confidence interval 95% (1.19–1.44); P=9.74 × 10−9).
doi:10.1038/tp.2014.2
PMCID: PMC3944635  PMID: 24495969
dementia risk; DNA; GWAS; molecular epidemiology; SNP; thyroid receptor
15.  Population Structure of the Endangered Franciscana Dolphin (Pontoporia blainvillei): Reassessing Management Units 
PLoS ONE  2014;9(1):e85633.
Franciscanas are the most endangered dolphins in the Southwestern Atlantic. Due to their coastal and estuarine habits, franciscanas suffer from extensive fisheries bycatch, as well as from habitat loss and degradation. Four Franciscana Management Areas (FMA), proposed based on biology, demography, morphology and genetic data, were incorporated into management planning and in the delineation of research efforts. We re-evaluated that proposal through the analysis of control region sequences from franciscanas throughout their distribution range (N = 162), including novel sequences from the northern limit of the species and two other previously unsampled localities in Brazil. A deep evolutionary break was observed between franciscanas from the northern and southern portions of the species distribution, indicating that they must be managed as two Evolutionarily Significant Units (ESU). Furthermore, additional FMAs should be recognised to accommodate the genetic differentiation found in each ESU. These results have immediate consequences for the conservation and management of this endangered species.
doi:10.1371/journal.pone.0085633
PMCID: PMC3908959  PMID: 24497928
16.  Correction: Effect of Insulin on ACE2 Activity and Kidney Function in the Non-Obese Diabetic Mouse 
PLoS ONE  2014;9(1):10.1371/annotation/2a7b69d2-a049-448e-b584-51e4c1e8a50d.
doi:10.1371/annotation/2a7b69d2-a049-448e-b584-51e4c1e8a50d
PMCID: PMC3905978
17.  Domestic Dogs in Rural Communities around Protected Areas: Conservation Problem or Conflict Solution? 
PLoS ONE  2014;9(1):e86152.
Although domestic dogs play many important roles in rural households, they can also be an important threat to the conservation of wild vertebrates due to predation, competition and transmission of infectious diseases. An increasing number of studies have addressed the impact of dogs on wildlife but have tended to ignore the motivations and attitudes of the humans who keep these dogs and how the function of dogs might influence dog-wildlife interactions. To determine whether the function of domestic dogs in rural communities influences their interactions with wildlife, we conducted surveys in rural areas surrounding protected lands in the Valdivian Temperate Forests of Chile. Sixty percent of farm animal owners reported the use of dogs as one of the primary means of protecting livestock from predators. The probability of dog–wild carnivore interactions was significantly associated with the raising of poultry. In contrast, dog–wild prey interactions were not associated with livestock presence but had a significant association with poor quality diet as observed in previous studies. Dog owners reported that they actively encouraged the dogs to chase off predators, accounting for 25–75% of the dog–wild carnivore interactions observed, depending on the predator species. Humans controlled the dog population by killing pups and unwanted individuals resulting in few additions to the dog population through breeding; the importation of predominantly male dogs from urban areas resulted in a sex ratios highly dominated by males. These results indicate that dog interactions with wildlife are related to the role of the dog in the household and are directly influenced by their owners. To avoid conflict with local communities in conservation areas, it is important to develop strategies for managing dogs that balance conservation needs with the roles that dogs play in these rural households.
doi:10.1371/journal.pone.0086152
PMCID: PMC3896434  PMID: 24465930
18.  Changes in Bacterial Population of Gastrointestinal Tract of Weaned Pigs Fed with Different Additives 
BioMed Research International  2014;2014:269402.
This study aimed to provide novel insights into the gastrointestinal microbial diversity from different gastrointestinal locations in weaning piglets using PCR-restriction fragment length polymorphism (PCR-RFLP). Additionally, the effect of different feed additives was analyzed. Thirty-two piglets were fed with four different diets: a control group and three enriched diets, with avilamycin, sodium butyrate, and a plant extract mixture. Digesta samples were collected from eight different gastrointestinal segments of each animal and the bacterial population was analysed by a PCR-RFLP technique that uses 16S rDNA gene sequences. Bacterial diversity was assessed by calculating the number of bands and the Shannon-Weaver index. Dendrograms were constructed to estimate the similarity of bacterial populations. A higher bacterial diversity was detected in large intestine compared to small intestine. Among diets, the most relevant microbial diversity differences were found between sodium butyrate and plant extract mixture. Proximal jejunum, ileum, and proximal colon were identified as those segments that could be representative of microbial diversity in pig gut. Results indicate that PCR-RFLP technique allowed detecting modifications on the gastrointestinal microbial ecology in pigs fed with different additives, such as increased biodiversity by sodium butyrate in feed.
doi:10.1155/2014/269402
PMCID: PMC3915759  PMID: 24575403
19.  Antibiotic Resistance Determinants in a Pseudomonas putida Strain Isolated from a Hospital 
PLoS ONE  2014;9(1):e81604.
Environmental microbes harbor an enormous pool of antibiotic and biocide resistance genes that can impact the resistance profiles of animal and human pathogens via horizontal gene transfer. Pseudomonas putida strains are ubiquitous in soil and water but have been seldom isolated from humans. We have established a collection of P. putida strains isolated from in-patients in different hospitals in France. One of the isolated strains (HB3267) kills insects and is resistant to the majority of the antibiotics used in laboratories and hospitals, including aminoglycosides, ß-lactams, cationic peptides, chromoprotein enediyne antibiotics, dihydrofolate reductase inhibitors, fluoroquinolones and quinolones, glycopeptide antibiotics, macrolides, polyketides and sulfonamides. Similar to other P. putida clinical isolates the strain was sensitive to amikacin. To shed light on the broad pattern of antibiotic resistance, which is rarely found in clinical isolates of this species, the genome of this strain was sequenced and analysed. The study revealed that the determinants of multiple resistance are both chromosomally-borne as well as located on the pPC9 plasmid. Further analysis indicated that pPC9 has recruited antibiotic and biocide resistance genes from environmental microorganisms as well as from opportunistic and true human pathogens. The pPC9 plasmid is not self-transmissible, but can be mobilized by other bacterial plasmids making it capable of spreading antibiotic resistant determinants to new hosts.
doi:10.1371/journal.pone.0081604
PMCID: PMC3894933  PMID: 24465371
21.  Effect of Insulin on ACE2 Activity and Kidney Function in the Non-Obese Diabetic Mouse 
PLoS ONE  2014;9(1):e84683.
We studied the non-obese diabetic (NOD) mice model because it develops autoimmune diabetes that resembles human type 1 diabetes. In diabetic mice, urinary albumin excretion (UAE) was ten-fold increased at an “early stage” of diabetes, and twenty-fold increased at a “later stage” (21 and 40 days, respectively after diabetes diagnosis) as compared to non-obese resistant controls. In NOD Diabetic mice, glomerular enlargement, increased glomerular filtration rate (GFR) and increased blood pressure were observed in the early stage. In the late stage, NOD Diabetic mice developed mesangial expansion and reduced podocyte number. Circulating and urine ACE2 activity were markedly increased both, early and late in Diabetic mice. Insulin administration prevented albuminuria, markedly reduced GFR, blood pressure, and glomerular enlargement in the early stage; and prevented mesangial expansion and the reduced podocyte number in the late stage of diabetes. The increase in serum and urine ACE2 activity was normalized by insulin administration at the early and late stages of diabetes in Diabetic mice. We conclude that the Diabetic mice develops features of early kidney disease, including albuminuria and a marked increase in GFR. ACE2 activity is increased starting at an early stage in both serum and urine. Moreover, these alterations can be completely prevented by the chronic administration of insulin.
doi:10.1371/journal.pone.0084683
PMCID: PMC3882249  PMID: 24400109
22.  Loss of Olfactory Receptor Function in Hominin Evolution 
PLoS ONE  2014;9(1):e84714.
The mammalian sense of smell is governed by the largest gene family, which encodes the olfactory receptors (ORs). The gain and loss of OR genes is typically correlated with adaptations to various ecological niches. Modern humans have 853 OR genes but 55% of these have lost their function. Here we show evidence of additional OR loss of function in the Neanderthal and Denisovan hominin genomes using comparative genomic methodologies. Ten Neanderthal and 8 Denisovan ORs show evidence of loss of function that differ from the reference modern human OR genome. Some of these losses are also present in a subset of modern humans, while some are unique to each lineage. Morphological changes in the cranium of Neanderthals suggest different sensory arrangements to that of modern humans. We identify differences in functional olfactory receptor genes among modern humans, Neanderthals and Denisovans, suggesting varied loss of function across all three taxa and we highlight the utility of using genomic information to elucidate the sensory niches of extinct species.
doi:10.1371/journal.pone.0084714
PMCID: PMC3879314  PMID: 24392153
23.  The relationship between executive functions and fluid intelligence in schizophrenia 
An enduring question is unity vs. separability of executive deficits resulting from impaired frontal lobe function. In previous studies, we have asked how executive deficits link to a conventional measure of fluid intelligence, obtained either by standard tests of novel problem-solving, or by averaging performance in a battery of novel tasks. For some classical executive tasks, such as the Wisconsin Card Sorting Test (WCST), Verbal Fluency, and Trail Making Test B (TMTB), frontal deficits are entirely explained by fluid intelligence. However, on a second set of executive tasks, including tests of multitasking and decision making, deficits exceed those predicted by fluid intelligence loss. In this paper we discuss how these results shed light on the diverse clinical phenomenology observed in frontal dysfunction, and present new data on a group of 15 schizophrenic patients and 14 controls. Subjects were assessed with a range of executive tests and with a general cognitive battery used to derive a measure of fluid intelligence. Group performance was compared and fluid intelligence was introduced as a covariate. In line with our previous results, significant patient-control differences in classical executive tests were removed when fluid intelligence was introduced as a covariate. However, for tests of multitasking and decision making, deficits remained. We relate our findings to those of previous factor analytic studies describing a single principal component, which accounts for much of the variance of schizophrenic patients' cognitive performance. We propose that this general factor reflects low fluid intelligence capacity, which accounts for much but not all cognitive impairment in this patient group. Partialling out the general effects of fluid intelligence, we propose, may clarify the role of additional, more specific cognitive impairments in conditions such as schizophrenia.
doi:10.3389/fnbeh.2014.00046
PMCID: PMC3932409  PMID: 24605092
executive function; fluid intelligence; schizophrenia; frontal lobe; multitasking; decision making
24.  Reduced striatal ecto-nucleotidase activity in schizophrenia patients supports the “adenosine hypothesis” 
Purinergic Signalling  2013;9(4):599-608.
Schizophrenia (SZ) is a major chronic neuropsychiatric disorder characterized by a hyperdopaminergic state. The hypoadenosinergic hypothesis proposes that reduced extracellular adenosine levels contribute to dopamine D2 receptor hyperactivity. ATP, through the action of ecto-nucleotidases, constitutes a main source of extracellular adenosine. In the present study, we examined the activity of ecto-nucleotidases (NTPDases, ecto-5′-nucleotidase, and alkaline phosphatase) in the postmortem putamen of SZ patients (n = 13) compared with aged-matched controls (n = 10). We firstly demonstrated, by means of artificial postmortem delay experiments, that ecto-nucleotidase activity in human brains was stable up to 24 h, indicating the reliability of this tissue for these enzyme determinations. Remarkably, NTPDase-attributable activity (both ATPase and ADPase) was found to be reduced in SZ patients, while ecto-5′-nucleotidase and alkaline phosphatase activity remained unchanged. In the present study, we also describe the localization of these ecto-enzymes in human putamen control samples, showing differential expression in blood vessels, neurons, and glial cells. In conclusion, reduced striatal NTPDase activity may contribute to the pathophysiology of SZ, and it represents a potential mechanism of adenosine signalling impairment in this illness.
doi:10.1007/s11302-013-9370-7
PMCID: PMC3889387  PMID: 23771238
Ecto-nucleotidases; Adenosine; CD39; CD73; Schizophrenia; Postmortem
25.  A mathematical model of serotype replacement in pneumococcal carriage following vaccination 
A number of childhood vaccination programmes have recently introduced vaccination against Streptococcus pneumoniae, the pneumococcus, a major cause of pneumonia and meningitis. The pneumococcal conjugate vaccines (PCVs) that are currently in use only protect against some serotypes of the bacterium, and there is now strong evidence that those serotypes not included in the vaccine increase in prevalence among most vaccinated populations. We present a mathematical model for the dynamics of nasopharyngeal carriage of S. pneumoniae that allows for carriage with multiple serotypes. The model is used to predict the prevalence of vaccine type (VT) and non-VT (NVT) serotypes following the introduction of PCV. Parameter estimates for the model are obtained by maximum likelihood using pre-vaccination data from The Gambia. The model predicts that low (1, 6A and 9V) and medium (4, 5, 7F, 14, 18C, 19A and 19F) prevalence serotypes can be eliminated through vaccination, but that the overall prevalence of carriage will be reduced only slightly because of an increase in the prevalence of NVT serotypes. Serotype replacement will be sequential, with high and medium prevalence NVT serotypes dominating initially, followed by an increase of serotypes of low prevalence. We examine the impact of a hypothetical vaccine that provides partial protection against all serotypes, and find that this reduces overall carriage, but is unable to eliminate low or medium prevalence serotypes.
doi:10.1098/rsif.2013.0786
PMCID: PMC3808555  PMID: 24132203
serotype replacement; bacterial carriage; mathematical model; pneumococcal vaccine

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