Development of a real-time PCR for Bartonella spp. detection, a current emerging microorganism.

A real-time PCR assay using SYBR Green was optimized to detect those Bartonella that are most frequently described as pathogens. The assay was genus-specific. Sequencing allowed to distinguish species. Assay sensitivity was determined using 10-fold serial dilutions of genomic DNA. Dynamic range was 100 ng-100 fg and sensitivity was 50 copies/reaction.

Severity of dementia, anosognosia, and depression in relation to the quality of life of patients with Alzheimer disease: discrepancies between patients and caregivers.

OBJECTIVE: To investigate the factors associated with discrepancies between patient and caregiver reports of the quality of life of patients (QoLp) with Alzheimer disease. METHODS: Cross-sectional analytic study of 141 patients and their caregivers. The instruments used were the Quality of Life in AD, the Global Deterioration Scale (GDS), the Geriatric Depression Scale, and the Anosognosia Questionnaire-Dementia. Differences were analyzed according to GDS stage. A linear regression analysis was conducted using the difference between the absolute QoLp scores of patients and caregivers. A cluster analysis involving patient variables was then performed. RESULTS: The discrepancy between patient and caregiver QoLp ratings increased in line with GDS stages (χ(2) (2) = 8.7, p = 0.013). In the regression model (F [7,133] = 16.6, p <0.001; R(2) = 0.477), discrepancies in QoLp reports were associated with greater anosognosia, less depression, and a better cognitive status in patients and with female gender among caregivers. The cluster analysis showed that patients with the lowest ratings of QoLp had a better cognitive status, more depression, and less anosognosia. Conversely, the highest ratings were given by patients with a poorer cognitive status, less depression, and greater anosognosia. CONCLUSIONS: The factors associated with greater discrepancies between patient and caregiver ratings of QoLp were severity of dementia, anosognosia, depression, and cognitive status in patients and female gender in caregivers. In patients with advanced dementia, greater anosognosia leads to more positive ratings in QoLp and complementary observations are required.

Understanding language and cognition after brain surgery - Tumour grade, fine-grained assessment tools and, most of all, individualized approach.

Cognitive performance influences the quality of life and survival of people with glioma. Thus, a detailed neuropsychological and language evaluation is essential. In this work, we tested if an analysis of errors in naming can indicate semantic and/or phonological impairments in 87 awake brain surgery patients. Secondly, we explored how language and cognition change after brain tumour resection. Finally, we checked if low-tumour grade had a protective effect on cognition. Our results indicated that naming errors can be useful to monitor semantic and phonological processing, as their number correlated with scores on tasks developed by our team for testing these domains. Secondly, we showed that - although an analysis at a whole group level indicates a decline in language functions - significantly more individual patients improve or remain stable when compared to the ones who declined. Finally, we observed that having LGG, when compared with HGG, favours patients' outcome after surgery, most probably due to brain plasticity mechanisms. We provide new evidence of the importance of applying a broader neuropsychological assessment and an analysis of naming errors in patients with glioma. Our approach may potentially ensure better detection of cognitive deficits and contribute to better postoperative outcomes. Our study also shows that an individualized approach in post-surgical follow-ups can reveal reassuring results showing that significantly more patients remain stable or improve and can be a promising avenue for similar reports. Finally, the study captures that plasticity mechanisms may act as protective in LGG versus HGG after surgery.

Cost-Effectiveness of Low-Field Intraoperative Magnetic Resonance in Glioma Surgery.

OBJECT: Low-field intraoperative magnetic resonance (LF-iMR) has demonstrated a slight increase in the extent of resection of intra-axial tumors while preserving patient`s neurological outcomes. However, whether this improvement is cost-effective or not is still matter of controversy. In this clinical investigation we sought to evaluate the cost-effectiveness of the implementation of a LF-iMR in glioma surgery. METHODS: Patients undergoing LF-iMR guided glioma surgery with gross total resection (GTR) intention were prospectively collected and compared to an historical cohort operated without this technology. Socio-demographic and clinical variables (pre and postoperative KPS; histopathological classification; Extent of resection; postoperative complications; need of re-intervention within the first year and 1-year postoperative survival) were collected and analyzed. Effectiveness variables were assessed in both groups: Postoperative Karnofsky performance status scale (pKPS); overall survival (OS); Progression-free survival (PFS); and a variable accounting for the number of patients with a greater than subtotal resection and same or higher postoperative KPS (R-KPS). All preoperative, procedural and postoperative costs linked to the treatment were considered for the cost-effectiveness analysis (diagnostic procedures, prosthesis, operating time, hospitalization, consumables, LF-iMR device, etc). Deterministic and probabilistic simulations were conducted to evaluate the consistency of our analysis. RESULTS: 50 patients were operated with LF-iMR assistance, while 146 belonged to the control group. GTR rate, pKPS, R-KPS, PFS, and 1-year OS were respectively 13,8% (not significative), 7 points (p < 0.05), 17% (p < 0.05), 38 days (p < 0.05), and 3.7% (not significative) higher in the intervention group. Cost-effectiveness analysis showed a mean incremental cost per patient of 789 € in the intervention group. Incremental cost-effectiveness ratios were 111 € per additional point of pKPS, 21 € per additional day free of progression, and 46 € per additional percentage point of R-KPS. CONCLUSION: Glioma patients operated under LF-iMR guidance experience a better functional outcome, higher resection rates, less complications, better PFS rates but similar life expectancy compared to conventional techniques. In terms of efficiency, LF-iMR is very close to be a dominant technology in terms of R-KPS, PFS and pKPS.

Detection of Rickettsia massiliae/Bar29 and Rickettsia conorii in red foxes (Vulpes vulpes) and their Rhipicephalus sanguineus complex ticks.

To determine the prevalence of exposure to Rickettsia massiliae/Bar29 and Rickettsia conorii in wild red foxes, we collected blood samples and ticks from 135 foxes shot in different game reserve areas in Catalonia. To detect SFG rickettsia in Rhipicephalus sanguineus complex ticks collected from the foxes, we used real-time polymerase chain reaction (PCR) to screen for ompA gene and a tick-borne bacteria flow chip technique based on multiplex PCR. Serum samples were positive for antibodies against spotted fever group (SFG) rickettsiae in 68 (50.3%). Molecular techniques identified R. massiliae in 107 ticks, R. aeschlimannii in 3 ticks, and R. slovaca in one tick; no R. conorii was identified in any of the ticks analyzed. We conclude that red foxes can carry ticks with SFG rickettsia.

Evaluation of the presence of Rickettsia slovaca infection in domestic ruminants in Catalonia, Northeastern Spain.

Rickettsia slovaca is the etiological agent of the human disease tick-borne lymphadenopathy (TIBOLA) transmitted by Dermacentor spp. ticks. In our area, Dermacentor marginatus is the most important tick vector; adult ticks feed on mammals, especially ungulates such as wild boars and domestic ruminants. The epidemiology of tick-transmitted diseases describes a wild cycle and a domestic cycle and both are connected by ticks. To identify the role of domestic ruminants in the transmission and maintenance of R. slovaca infection, blood samples from sheep (n=95), goats (n=91), and bullfighting cattle (n=100) were collected during a herd health program, and livestock grazing was selected to ensure tick contact. Samples were analyzed by serology using an indirect immunofluorescent assay (IFA) and molecular techniques (real-time PCR). Seroprevalence was 15.7% in sheep, 20.8% in goats, and 65.0% in bullfighting cattle. On the basis of molecular methods, R. slovaca infection was demonstrated in a goat blood sample with an antibody titer of 1:160. This is the first time that R. slovaca has been identified in a goat blood sample. These results suggest that domestic ruminants are exposed to R. slovaca infection and, because the domestic cycle is close to the human environment, this could increase the risk of transmitting the pathogen to human beings.

Rickettsia slovaca infection in humans in the northeast of Spain: seroprevalence study.

Catalonia is an endemic area of Mediterranean spotted fever. In 1997, A. Lakos described a new tick-borne infectious disease called tick-borne lymphadenopathy. The causative agent is Rickettsia slovaca, which is transmitted by Dermacentor marginatus ticks. We have diagnosed human cases in Catalonia. The objective of this study was to determinate seroprevalence of R. slovaca infection in humans in the northeast of Spain. The population included 217 subjects from Catalonia, northeast of Spain and was stratified by age and living place (rural, suburban, and urban). Age, gender, residence area, contact with animals, occupation, and history of rickettsioses was surveyed. Immunoglobulin G was measured by indirect immunofluorescence assay. Titers >or= 1/40 were considered. Seroprevalence of R. slovaca was 5.5% at titers of 1/40-1/320. Eight (3.7%) sera had antibodies against R. slovaca exclusively. Four sera reacted also against Rickettsia conorii and/or Bar29. Seroprevalence of R. slovaca would range from 3.7% to 5.5%. The only statistically significant association was that between R. slovaca seropositivity and age. We present serologic evidence of R. slovaca infection among population of Catalonia, northeast of Spain.