Comparison of the whole slide imaging and conventional light microscopy in the grading of oral epithelial dysplasia: a multi-institutional study.

BACKGROUND: Whole Slide Imaging (WSI) is an alternative method to light microscopy (LM). However, few studies have compared the diagnostic agreement between WSI and LM, especially to grade oral epithelial dysplasia (OED). The purpose of this study was to evaluate the variability in grading OED by the World Health Organization grading system, using WSI and conventional LM, and to investigate whether the access to clinical information, and psychologic or physical states of the pathologists could interfere with the diagnosis. MATERIAL AND METHODS: eleven experienced pathologists from seven Brazilian universities independently evaluated twenty-five OED cases. The analyses were performed in duplicate for each method, with an interval of at least 30 days, and the time consumed in each analysis was measured. Physical and psychologic states were evaluated by blood pressure levels, heart rate and two questionnaires: State-Trait Anxiety Inventory and Perceived Stress Scale. Clinical information was provided after the second evaluation using WSI and the pathologist could change their diagnostic decision or not. RESULTS: LM showed a higher inter-examiner agreement (k=0.53) than WSI (k=0.45) and a smaller time consumed by the pathologists (mean of 65.53 seconds compared to 91.02 seconds in WSI). In the first analysis using conventional microscopy, there was a positive correlation between kappa values and anxiety (r=0.47, p=0.02), and stress (r=0.64, p<0.01), and an inverse correlation with heart rate (r=-0.48, p=0.02). In the digital analysis, there was also a positive correlation between kappa values and anxiety (r=0.75, p<0.001). After clinical information was given, there was a slight change in 11.3% of the cases, and a great discrepancy in 1.1% of the cases, mainly increasing the OED grade. CONCLUSIONS: both microscopy systems had similar results, although LM had slightly higher kappa values, and WSI was more time consuming.

Children's public health: Danger of exposure to pathogenic fungi in recreational places in the middle-west region of Brazil.

BACKGROUND: Dermatophytes are keratinophilic fungi, considered etiological agents of cutaneous mycoses in man and animals. The objective of this work was to isolate dermatophytic and non-dermatophytic fungi in recreational sandboxes (rainy and dry periods) in public day-care centers located in the city of Cuiabá-MT (Middle-west of Brazil). METHODS: Samples (n=200) were collected from the superficial layer of the sandboxes at a depth of 2-5cm in 10 public nurseries in Cuiabá-MT. Hair baits measuring 1-2cm were autoclaved and dispersed in 50g of the sample in sterile plates, incubated at 28°C, moistened with chloramphenicol solution (50mg/L). After 6 weeks the baits were inoculated on Sabouraud agar plus 50mg/L chloramphenicol and 500mg/L cycloheximide. The fungi were identified according to macroscopic and micromorphological characteristics. RESULTS: From the collected sand samples, 1318 colonies and 56 species of fungi belonging to 22 genera were isolated. The most important genera were Paecilomyces spp. (30.42%), Penicillium spp. (19.12%), Fusarium spp. (11.46%) and Aspergillus spp. (11.15%). Dermatophytes were recovered in 50% of day-care centers in a total of 29 identified colonies, the Trichophyton genus (86.2%) being the most frequently isolated. CONCLUSIONS: The dermatophytes in the recreational areas of day-care centers can pose a risk to the health of its users, especially for children and the employees. Work with this profile is very important to guide actions related to health surveillance.

Anatomical and Functional MRI Changes after One Year of Auditory Rehabilitation with Hearing Aids.

Hearing aids (HAs) are an effective strategy for auditory rehabilitation in patients with peripheral hearing deficits. Yet, the neurophysiological mechanisms behind HA use are still unclear. Thus far, most studies have focused on changes in the auditory system, although it is expected that hearing deficits affect a number of cognitive systems, notably speech. In the present study, we used audiometric evaluations in 14 patients with bilateral hearing loss before and after one year of continuous HA use and functional magnetic resonance imaging (fMRI) and cortical thickness analysis in 12 and 10 of them compared with a normal hearing control group. Prior to HA fitting, fMRI activity was found reduced in the auditory and language systems and increased in visual and frontal areas, expanding to multimodal integration cortices, such as the superior temporal gyrus, intraparietal sulcus, and insula. One year after rehabilitation with HA, significant audiometric improvement was observed, especially in free-field Speech Reception Threshold (SRT) test and functional gain, a measure of HA efficiency. HA use increased fMRI activity in the auditory and language cortices and multimodal integration areas. Individual fMRI signal changes from all these areas were positively correlated with individual SRT changes. Before rehabilitation, cortical thickness was increased in parts of the prefrontal cortex, precuneus, fusiform gyrus, and middle temporal gyrus. It was reduced in the insula, supramarginal gyrus, medial temporal gyrus, occipital cortex, posterior cingulate cortex, and claustrum. After HA use, increased cortical thickness was observed in multimodal integration regions, particularly the very caudal end of the superior temporal sulcus, the angular gyrus, and the inferior parietal gyrus/superior temporal gyrus/insula. Our data provide the first evidence that one year of HA use is related to functional and anatomical brain changes, notably in auditory and language systems, extending to multimodal cortices.

Doxycycline treatment efficacy in dogs with naturally occurring Anaplasma phagocytophilum infection.

OBJECTIVES: To evaluate doxycycline treatment efficacy and post-treatment pathogen persistence in dogs naturally infected with Anaplasma phagocytophilum in endemic regions of the USA. MATERIALS AND METHODS: Symptomatic dogs in four US states (MN, WI, CT and CA) were evaluated before treatment with doxycycline and approximately 30 and 60 days post-treatment. Clinicopathological parameters, co-exposures and A. phagocytophilum DNA in whole blood and lymph node samples were compared between A. phagocytophilum infected and uninfected dogs. RESULTS: In total, 42 dogs fulfilled the inclusion criteria, with 16 dogs (38%) blood PCR-positive and 26 dogs (62%) blood PCR-negative for A. phagocytophilum. At initial evaluation, the proportion of clinicopathological abnormalities was similar between A. phagocytophilum infected and uninfected dogs, although thrombocytopenia and lymphopenia were statistically more prevalent among A. phagocytophilum infected dogs. Treatment with doxycycline resulted in resolution of all clinical abnormalities in infected dogs; four dogs had persistent haematological abnormalities, including mild leukopenia, eosinopenia and lymphopenia. All 16 infected dogs became blood PCR-negative approximately 30 and 60 days after treatment onset. Additionally, 13/13 (100%) lymph node specimens tested post-treatment were PCR-negative. Select clinicopathological abnormalities persisted in uninfected dogs after treatment. CLINICAL SIGNIFICANCE: The results of this study support the efficacy of doxycycline therapy for clinical treatment of dogs naturally infected with A. phagocytophilum in the USA. This study did not find clinical, haematological or microbiological indicators that supported the persistence of A. phagocytophilum infection in naturally infected dogs following treatment with doxycycline for 28 days.

Percutaneous hallux valgus treatment: Unilaterally or bilaterally.

INTRODUCTION: Currently there is no consensus regarding the use of bilateral simultaneous percutaneous surgery for Hallux valgus treatment. Although the technique described in M. Prado's book, recommends operating only one foot at a time there are no published studies confirming it. The aim of this study was to evaluate whether there is a difference between the results of patients that have been percutaneously operated on one foot and those operated on both feet at the same surgical time for mild to moderate Hallux valgus correction. MATERIAL/METHODS: We did a retrospective single centre evaluation of 93 feet (61 patients) with Hallux valgus operated percutaneously. 29 patients were operated unilaterally (group I) and 32 bilaterally simultaneously (group II) between 2005 and 2009. The Metatarsophalangeal angle (MPA), Intermetatarsal angle (IMA) and Distal metatarsal articular angle (DMAA) were evaluated pre- and postoperatively. The AOFAS score, the degree of patients' satisfaction and the complications were evaluated postoperatively. RESULTS: The mean follow-up was 24.0 months in group I, and 28.0 in group II. The average postoperative AOFAS score was 86.8, 82.9 in group I and 88.6 in group II (p>0.05). 90.6% were satisfied or very satisfied in group I, and 89.7% in group II (p>0.05). There was no statistically significant difference (p>0.05) in the average correction of MPA, DMAA, and IMA. The complications rate was similar in both groups. CONCLUSIONS: The similar results obtained in both groups suggest that the simultaneous bilateral percutaneous surgery gives equivalent results to the unilateral, which has an important socioeconomic impact since there is only one recovery time for both feet. Further research is needed.

Effects of oocyte donor age and embryonic stage of development on transcription of genes coding for enzymes of the prostaglandins and progesterone synthesis pathways in bovine in vitro produced embryos.

The ability of early bovine embryos to produce prostaglandins (PGs) and progesterone (P4), and the role of these mediators in embryonic development and survival are poorly understood. In this study we tested the hypothesis that day 7 bovine embryos are able to transcribe genes coding for enzymes of the PGs (PTGS2, PGES, PGFS) and P4 (StAR, P450scc, 3ß-HSD) synthesis pathways, and that transcription levels of these genes are associated with developmental progression and heifer age-related [pre-pubertal (PP) versus post-pubertal cyclic (C)] oocyte competence. Compared with C heifer oocytes, PP heifer oocytes showed a lower (P < 0.0001) in vitro blastocyst rate, but in embryos developing until day 7, heifer age had no effect on quality grade. Day 7 quality grade 1-2 embryos were selected for RNA extraction and gene transcription analysis by qRT-PCR, in a 2 × 2 factorial design [age (PP or C) × embryonic stage (compact morulae and early blastocysts, CM + EBL, or blastocysts and expanded blastocysts, BL + BEX); 15 embryos/group]. Transcription levels of PTGS2, PGES, PGFS, P450scc and 3ß-HSD were not affected by heifer age but were higher (P < 0.01) in BL + BEX than in CM + EBL. In conclusion, the main limiting factor for embryo production from PP heifers is oocyte competence. Day 7 bovine embryos evidence transcription of genes coding for enzymes of PGs and P4 synthesis pathways, and transcription levels are associated with blastocyst differentiation. This prompts for an autocrine/paracrine action of PGs and P4 in early bovine embryonic development.

Occult hepatitis B virus infection in liver transplant patients in a Brazilian referral center

Estimates of occult hepatitis B virus (HBV) infection prevalence varies among different studies depending on the prevalence of HBV infection in the study population and on the sensitivity of the assay used to detect HBV DNA. We investigated the prevalence of occult HBV infection in cirrhotic patients undergoing liver transplantation in a Brazilian referral center. Frozen liver samples from 68 adults were analyzed using a nested polymerase chain reaction assay for HBV DNA. The specificity of the amplified HBV sequences was confirmed by direct sequencing of the amplicons. The patient population comprised 49 (72.1%) males and 19 (27.9%) females with a median age of 53 years (range=18-67 years). Occult HBV infection was diagnosed in three (4.4%) patients. The etiologies of the underlying chronic liver disease in these cases were alcohol abuse, HBV infection, and cryptogenic cirrhosis. Two of the patients with cryptic HBV infection also presented hepatocellular carcinoma. Markers of previous HBV infection were available in two patients with occult HBV infection and were negative in both. In conclusion, using a sensitive nested polymerase chain reaction assay to detect HBV DNA in frozen liver tissue, we found a low prevalence of occult HBV infection in cirrhotic patients undergoing liver transplant, probably due to the low prevalence of HBV infection in our population.

Occult hepatitis B virus infection in liver transplant patients in a Brazilian referral center.

Estimates of occult hepatitis B virus (HBV) infection prevalence varies among different studies depending on the prevalence of HBV infection in the study population and on the sensitivity of the assay used to detect HBV DNA. We investigated the prevalence of occult HBV infection in cirrhotic patients undergoing liver transplantation in a Brazilian referral center. Frozen liver samples from 68 adults were analyzed using a nested polymerase chain reaction assay for HBV DNA. The specificity of the amplified HBV sequences was confirmed by direct sequencing of the amplicons. The patient population comprised 49 (72.1%) males and 19 (27.9%) females with a median age of 53 years (range=18-67 years). Occult HBV infection was diagnosed in three (4.4%) patients. The etiologies of the underlying chronic liver disease in these cases were alcohol abuse, HBV infection, and cryptogenic cirrhosis. Two of the patients with cryptic HBV infection also presented hepatocellular carcinoma. Markers of previous HBV infection were available in two patients with occult HBV infection and were negative in both. In conclusion, using a sensitive nested polymerase chain reaction assay to detect HBV DNA in frozen liver tissue, we found a low prevalence of occult HBV infection in cirrhotic patients undergoing liver transplant, probably due to the low prevalence of HBV infection in our population.

Evaluation of treatments with hCG and carprofen at embryo transfer in a demi-embryo and recipient virgin heifer model.

An in vivo model, combining a low developmental competence embryo (demi-embryo) and a high-fertility recipient (virgin dairy heifer) was used to evaluate the effects of treatment with human chorionic gonadotropin (hCG) and carprofen at embryo transfer (ET) on plasma progesterone (P4) concentrations of recipients and on embryonic growth and survival. Embryos were bisected and each demi-embryo was transferred to a recipient on Day 7 of the estrous cycle. At ET, heifers (n = 163) were randomly allocated to treatment with hCG (2500 IU im), carprofen (500 mg iv), hCG plus carprofen or to untreated controls. Plasma P4 concentrations were measured on Days 0, 7, 14 and 21 of all recipients plus on Days 28, 42 and 63 of pregnant recipients. Pregnancy was presumed to be present in recipients with luteal plasma P4 concentrations until Day 21 and confirmed by using transrectal ultrasonography on Days 28, 42 and 63. Embryonic measurements (crown-rump length and width) were obtained on Day 42. Treatment with hCG induced formation of secondary corpora lutea (CL) in 97% of heifers and increased (P < 0.01) mean plasma P4 concentrations of non-pregnant recipients on Day 14 and of pregnant heifers on Days 14 to 63. This was associated to a significant decrease in early embryonic mortality. In contrast, subsequent embryonic losses resulted in a non-significant numerical increase by 8% of pregnancies maintained to Day 63. Therefore, treatment with hCG significantly rescued embryos through the maternal recognition of pregnancy window but was not able to support development thereafter. Treatment with carprofen at ET had no significant effects on plasma P4 concentrations and rate of embryo mortality. Treatment with hCG plus carprofen at ET induced formation of secondary CL in 90% of heifers but decreased the luteotrophic effect of hCG, resulting in no effect on embryo survival. Low developmental competence embryos showed an intrinsic deficiency in overcoming the maternal recognition of pregnancy challenge and in proceeding to further development until Day 28 of pregnancy, whereas mortality beyond this point was residual. Results on pregnancy rates should be confirmed in further experiments involving a larger sample size.

Development of a bovine luteal cell in vitro culture system suitable for co-culture with early embryos.

The cross talk between the corpus luteum (CL) and the early embryo, potentially relevant to pregnancy establishment, is difficult to evaluate in the in vivo bovine model. In vitro co-culture of bovine luteal cells and early embryos (days 2-8 post in vitro fertilization) may allow the deciphering of this poorly understood cross talk. However, early embryos and somatic cells require different in vitro culture conditions. The objective of this study was to develop a bovine luteal cell in vitro culture system suitable for co-culture with early embryos in order to evaluate their putative steroidogenic and prostanoid interactions. The corpora lutea of the different stages of the estrous cycle (early, mid, and late) were recovered postmortem and enriched luteal cell populations were obtained. In experiments 1 and 2, the effects of CL stage, culture medium (TCM, DMEM-F12, or SOF), serum concentration (5 or 10%), atmosphere oxygen tension (5 or 20%), and refreshment of the medium on the ability of luteal cells to produce progesterone (P(4)) were evaluated. The production of P(4) was significantly increased in early CL cultures, and luteal cells adapted well to simple media (SOF), low serum concentrations (5%), and oxygen tensions (5%). In experiment 3, previous luteal cell cryopreservation did not affect the production of P(4), PGF(2α), and PGE(2) compared to fresh cell cultures. This enables the use of pools of frozen-thawed cells to decrease the variation in cell function associated with primary cell cultures. In experiment 4, mineral oil overlaying culture wells resulted in a 50-fold decrease of the P(4) quantified in the medium, but had no effect on PGF(2α) and PGE(2) quantification. In conclusion, a luteal cell in vitro culture system suitable for the 5-d-long co-culture with early embryos was developed.

Cryptococcus spp isolated from dust microhabitat in Brazilian libraries.

BACKGROUND: The Cryptococcus spp is currently composed of encapsulated yeasts of cosmopolitan distribution, including the etiological agents of cryptococcosis. The fungus are found mainly in substrates of animal and plant origin. Human infection occurs through inhalation of spores present in the environment. METHODS: Eighty-four swab collections were performed on dust found on books in three libraries in the city of Cuiabá, state of Mato Grosso, Brazil. The material was seeded in Sabouraud agar and then observed for characteristics compatible with colonies with a creamy to mucous aspect; the material was then isolated in birdseed (Niger) agar and cultivated at a temperature of 37°C for 5 to 7 days. Identification of isolated colonies was performed by microscopic observation in fresh preparations dyed with India ink, additional tests performed on CGB (L-canavanine glycine bromothymol blue), urea broth, and carbohydrate assimilation tests (auxanogram). RESULTS: Of the 84 samples collected from book dust, 18 (21.4%) were positive for Cryptococcus spp totalizing 41 UFC's. The most frequently isolated species was C. gattii 15 (36.6%); followed by C. terreus, 12 (29.3%); C. luteolus 4 (9.8%); C. neoformans, and C. uniguttulatus 3 (7.3%), and C. albidus and C. humiculus with 2 (4.6%) of the isolates. CONCLUSION: The high biodiversity of the yeasts of the Cryptococcus genus, isolated from different environmental sources in urban areas of Brazil suggests the possibility of individuals whose immune systems have been compromised or even healthy individuals coming into sources of fungal propagules on a daily bases throughout their lives. This study demonstrates the acquisition possible of cryptococcosis infection from dust in libraries.

Avaliação dos resultados e da satisfação global após tratamento da disfunção eréctil com prótese peniana insuflável de três componentes: a nossa experiência / Outcome and satisfaction rates of three-piece penile prostheses implantation for treatment of erectile dysfunction: single center experience

Introdução e objectivos: A colocação de prótese peniana é a última opção no tratamento da disfunção eréctil orgânica, quando todos os tratamentos de primeira e segunda linha não estão indicados ou são ineficazes. O objectivo deste estudo foi a avaliação dos doentes tratados com prótese peniana insuflável de três componentes e o seu impacto de satisfação no doente-parceira no nosso serviço, nos últimos 10 anos. Material e métodos: Entre 2001 e 2010, foram colocadas quarenta e uma (41) próteses penianas insufláveis de 3 componentes em homens com disfunção eréctil. Dois doentes não foram localizados na altura do estudo. Os dados foram obtidos nos registos clínicos, avaliação médica e entrevista telefónica. O casal preencheu um questionário de avaliação da qualidade de vida e satisfação global. A idade média na altura da cirurgia foi de 59,6 anos (38-73 anos). O tempo médio de seguimento após a cirurgia foi de 18 meses (6-77 meses). Resultados: As principais causas da disfunção eréctil foram antecedente cirúrgico de prostatectomia radical (46,4%) e a diabetes mellitus (39%). Seis doentes (14,6%) tiveram complicações pós-operatórias minor precoces. Em 4 doentes (9,7%) a prótese foi removida por infecção e não reimplantada. Não foi constatada disfunção mecânica ou necessi-dade de revisão das próteses. Na maioria dos doentes (94,6%) houve melhoria da libido e no relacionamento com as parceiras. Conclusão: O grau de satisfação global dos doentes e das parceiras após a colocação da prótese foi elevado, tendo-se registado apenas um pequeno número de complicações. Estes resultados demonstram que a colocação de prótese peniana insuflável de três componentes continua a ser o tratamento de eleição da disfunção eréctil orgânica refractária ao tratamento de primeira linha (AU)

Introduction and objective: Three-piece inflatable penile prostheses are the last option in the treatment of organic erectile dysfunction, when conservative therapies are unsuccessful, contraindicated or rejected. Our objective was to investigate the ten year outcome of patients treated with three-piece inflatable implant and its impact on patient-partner satisfaction in our department. Material and methods: Since 2001 to 2010, 41 patients with erectile dysfunction received a three piece penile implant. Two patients were lost for follow-up. Data was obtained from medical records, clinical evaluation and telephone interview. All patients and partners were assessed with a questionnaire about life quality and satisfaction. Mean age at surgery was 59.6 years (38-73 yrs). Median follow-up time was of 18 months (6-77 months). Results: Radical prostatectomy (46.4%) and diabetes mellitus (39.0%) were the main reason for dysfunction in this patient group. Six patients (14.6%) had early postoperative minor complications. Four patients (9.7%) presented with penile implant infection and the prosthesis were removed and not re-implanted. There were no reports of mechanical failure or prosthesis revision. The majority of patients (94.6%) were satisfied after surgery, with significant improvement in libido and relationship with partners. Conclusion: The global satisfaction of the patients and their partners after three-piece penile inflatable implantation is high, with a low complication rate. These results confirmed that three-piece inflatable penile prostheses are the gold standard in treatment of organic erectile dysfunction when conservative therapies failed (AU)

Extratemporal damage in temporal lobe epilepsy: magnetization transfer adds information to volumetric MR imaging.

BACKGROUND AND PURPOSE: MTS is characterized by gliosis and atrophy of the hippocampus and related limbic structures. However, the damage is not limited to those structures with atrophy and has been reported in extratemporal regions. Because volumetric studies are nonspecific, the pathophysiology of the brain damage remains to be solved. MTI is an MR imaging technique more sensitive to subtle neuropathologic changes than conventional MR imaging. Here we combined MTI with VBM analysis to evaluate extratemporal damage in patients with TLE. MATERIALS AND METHODS: We studied 23 healthy controls and 21 patients with TLE with mean ages, respectively, of 37.6 ± 10.9 and 38.6 ± 9.02 years. All subjects had a full clinical follow-up and MR imaging. We processed the images with VBM for volumetric analysis of WM and GM, as well as with voxel-based analysis of MTR for macromolecular integrity analysis. RESULTS: In addition to MTR decrease in the temporal lobes, we found a significant decrease in GM and WM volumes. In the WM, the MTR decrease was correlated to volume loss detected by VBM, indicating that brain atrophy may explain part of the MTR decrease. We also found areas in which the MTR decrease was not associated with volume loss, suggesting an additional pathophysiologic process other than neuronal loss and atrophy underlying the MTR changes. CONCLUSIONS: Our results support the hypothesis that there are widespread lesions in the brain, including the corpus callosum and the frontal lobe, affecting both GM and WM.

Molecular and serological diagnosis of Bartonella infection in 61 dogs from the United States.

BACKGROUND: Molecular diagnosis of canine bartonellosis can be extremely challenging and often requires the use of an enrichment culture approach followed by PCR amplification of bacterial DNA. HYPOTHESES: (1) The use of enrichment culture with PCR will increase molecular detection of bacteremia and will expand the diversity of Bartonella species detected. (2) Serological testing for Bartonella henselae and Bartonella vinsonii subsp. berkhoffii does not correlate with documentation of bacteremia. ANIMALS: Between 2003 and 2009, 924 samples from 663 dogs were submitted to the North Carolina State University, College of Veterinary Medicine, Vector Borne Diseases Diagnostic Laboratory for diagnostic testing with the Bartonella α-Proteobacteria growth medium (BAPGM) platform. Test results and medical records of those dogs were retrospectively reviewed. METHODS: PCR amplification of Bartonella sp. DNA after extraction from patient samples was compared with PCR after BAPGM enrichment culture. Indirect immunofluorescent antibody assays, used to detect B. henselae and B. vinsonii subsp. berkhoffii antibodies, were compared with PCR. RESULTS: Sixty-one of 663 dogs were culture positive or had Bartonella DNA detected by PCR, including B. henselae (30/61), B. vinsonii subsp. berkhoffii (17/61), Bartonella koehlerae (7/61), Bartonella volans-like (2/61), and Bartonella bovis (2/61). Coinfection with more than 1 Bartonella sp. was documented in 9/61 dogs. BAPGM culture was required for PCR detection in 32/61 cases. Only 7/19 and 4/10 infected dogs tested by IFA were B. henselae and B. vinsonii subsp. berkhoffii seroreactive, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs were most often infected with B. henselae or B. vinsonii subsp. berkhoffii based on PCR and enrichment culture, coinfection was documented, and various Bartonella species were identified. Most infected dogs did not have detectable Bartonella antibodies.

Prevention of endemic canine vector-borne diseases using imidacloprid 10% and permethrin 50% in young dogs: a longitudinal field study.

Canine vector-borne diseases (CVBDs) are highly prevalent and increasing in distribution worldwide. A longitudinal study was conducted in southern Italy to determine the incidence of and protection against CVBD-causing pathogens in dogs treated with a combination of imidacloprid 10% and permethrin 50% (ImPer). One hundred eleven autochthonous young dogs were divided into group A (n=63) and group B (n=48), both groups containing dogs positive and negative for one or more CVBD-causing pathogens. Additionally, 10 naïve male beagles were introduced in each group in May 2008. Group A was treated with ImPer on day 0 and every 21+/-2 days whereas group B was left untreated. Blood and skin samples were collected at baseline (March-April 2008) and at the first, second and third follow-up times (July and October 2008 and April 2009). Bone marrow was sampled at baseline and at the third follow-up. Serological, cytological and molecular tests were performed to detect Anaplasma platys, Babesia spp., Bartonella spp., Dirofilaria immitis, Ehrlichia canis, Hepatozoon canis and Leishmania infantum. Ectoparasites (fleas, ticks, and sand flies) were monitored throughout the study. The baseline prevalence of CVBDs was 39.6% with 44 dogs positive for at least one pathogen. A. platys (27.5%) and Babesia spp. (15.6%) were the most prevalent species and co-infections with up to two pathogens were detected in 16 (14.7%) individuals. At the end of the evaluation period, there was a 90.7% reduction in overall CVBD incidence density rate (IDR) in group A, as following: 100% reduction in L. infantum; 94.6% in E. canis; 94.4% in Babesia spp.; and 81.8% in A. platys. Initially positive treated dogs showed significantly lower pathogen prevalence at the third follow-up than untreated ones. At the end of the evaluation period, 8 of the 10 untreated beagles were infected with at least one pathogen whereas one of the treated beagles was A. platys positive at a single time point (second follow-up). Overall efficacy against ticks was 97.9%. In October 2009, samples were collected from the remaining 83 dogs (44 from group A and 39 from group B) to investigate the annual incidence of CVBDs in the same, at this time untreated, dog population. A high year incidence for tick-borne diseases (78.1%) and for L. infantum (13.6%) was detected in dogs from group A, seven months after the treatment had been withdrawn. The results demonstrate that ImPer preventive treatment against arthropods protects autochthonous and naïve beagle dogs against CVBD-causing pathogens.

Evaluation of brain tissue or cerebrospinal fluid with broadly reactive polymerase chain reaction for Ehrlichia, Anaplasma, spotted fever group Rickettsia, Bartonella, and Borrelia species in canine neurological diseases (109 cases).

BACKGROUND: Vector-transmitted microorganisms in the genera Ehrlichia, Anaplasma, Rickettsia, Bartonella, and Borrelia are commonly suspected in dogs with meningoencephalomyelitis (MEM), but the prevalence of these pathogens in brain tissue and cerebrospinal fluid (CSF) of dogs with MEM is unknown. HYPOTHESIS/OBJECTIVES: To determine if DNA from these genera is present in brain tissue and CSF of dogs with MEM, including those with meningoencephalitis of unknown etiology (MUE) and histopathologically confirmed cases of granulomatous (GME) and necrotizing meningoencephalomyelitis (NME). ANIMALS: Hundred and nine dogs examined for neurological signs at 3 university referral hospitals. METHODS: Brain tissue and CSF were collected prospectively from dogs with neurological disease and evaluated by broadly reactive polymerase chain reaction (PCR) for Ehrlichia, Anaplasma, Spotted Fever Group Rickettsia, Bartonella, and Borrelia species. Medical records were evaluated retrospectively to identify MEM and control cases. RESULTS: Seventy-five cases of MUE, GME, or NME, including brain tissue from 31 and CSF from 44 cases, were evaluated. Brain tissue from 4 cases and inflammatory CSF from 30 cases with infectious, neoplastic, compressive, vascular, or malformative disease were evaluated as controls. Pathogen nucleic acids were detected in 1 of 109 cases evaluated. Specifically, Bartonella vinsonii subsp. berkhoffii DNA was amplified from 1/6 dogs with histopathologically confirmed GME. CONCLUSION AND CLINICAL IMPORTANCE: The results of this investigation suggest that microorganisms in the genera Ehrlichia, Anaplasma, Rickettsia, and Borrelia are unlikely to be directly associated with canine MEM in the geographic regions evaluated. The role of Bartonella in the pathogenesis of GME warrants further investigation.

Brain tissue segmentation using q-entropy in multiple sclerosis magnetic resonance images

The loss of brain volume has been used as a marker of tissue destruction and can be used as an index of the progression of neurodegenerative diseases, such as multiple sclerosis. In the present study, we tested a new method for tissue segmentation based on pixel intensity threshold using generalized Tsallis entropy to determine a statistical segmentation parameter for each single class of brain tissue. We compared the performance of this method using a range of different q parameters and found a different optimal q parameter for white matter, gray matter, and cerebrospinal fluid. Our results support the conclusion that the differences in structural correlations and scale invariant similarities present in each tissue class can be accessed by generalized Tsallis entropy, obtaining the intensity limits for these tissue class separations. In order to test this method, we used it for analysis of brain magnetic resonance images of 43 patients and 10 healthy controls matched for gender and age. The values found for the entropic q index were 0.2 for cerebrospinal fluid, 0.1 for white matter and 1.5 for gray matter. With this algorithm, we could detect an annual loss of 0.98 percent for the patients, in agreement with literature data. Thus, we can conclude that the entropy of Tsallis adds advantages to the process of automatic target segmentation of tissue classes, which had not been demonstrated previously.

Brain tissue segmentation using q-entropy in multiple sclerosis magnetic resonance images.

The loss of brain volume has been used as a marker of tissue destruction and can be used as an index of the progression of neurodegenerative diseases, such as multiple sclerosis. In the present study, we tested a new method for tissue segmentation based on pixel intensity threshold using generalized Tsallis entropy to determine a statistical segmentation parameter for each single class of brain tissue. We compared the performance of this method using a range of different q parameters and found a different optimal q parameter for white matter, gray matter, and cerebrospinal fluid. Our results support the conclusion that the differences in structural correlations and scale invariant similarities present in each tissue class can be accessed by generalized Tsallis entropy, obtaining the intensity limits for these tissue class separations. In order to test this method, we used it for analysis of brain magnetic resonance images of 43 patients and 10 healthy controls matched for gender and age. The values found for the entropic q index were 0.2 for cerebrospinal fluid, 0.1 for white matter and 1.5 for gray matter. With this algorithm, we could detect an annual loss of 0.98% for the patients, in agreement with literature data. Thus, we can conclude that the entropy of Tsallis adds advantages to the process of automatic target segmentation of tissue classes, which had not been demonstrated previously.

Molecular documentation of Bartonella infection in dogs in Greece and Italy.

This study reports the occurrence of "Bartonella rochalimae" in Europe and the presence of Bartonella vinsonii subsp. berkhoffii genotypes II and III in dogs in southern Italy and provides DNA sequencing evidence of a potentially new Bartonella sp. infecting dogs in Greece and Italy.