Temporal summation of second pain is affected by cognitive load.

This work attempted to clarify the interaction of cognition and pain sensitization during a paradigm of Temporal Summation of Second Pain (TSSP). We analyzed pain ratings and electroencephalographic (EEG) activity obtained from 21 healthy participants during the presentation of four experimental conditions that differed in the manipulation of attention to painful stimuli or working memory load (Attention to hand & TSSP; 0-back & TSSP (low cognitive load); 2-back & TSSP (high cognitive load); 2-back (without pain)). We found that the TSSP was reduced when the attention was diverted and the cognitive load increased, and this reduction was accompanied by higher midfrontal theta activity and lower posterior alpha and central beta activity. Although it is well established that TSSP is a phenomenon that occurs at the spinal level, here we show that it is also affected by supraspinal attentional mechanisms. Delivery of painful repeated stimuli did not affect the performance of the 2-back task but was associated with smaller amplitudes of attentional event-related potentials (ERPs) after standard stimuli (not the target). The study of brain activity during TSSP allowed to clarify the role of top-down attentional modulation in pain sensitization processes. Results contribute to a better understanding of cognitive dysfunction in pain conditions and reinforce the use of therapeutic strategies based on distracting attention away from pain.

A family-based study to identify genetic biomarkers of fibromyalgia: consideration of patients' subgroups.

OBJECTIVES: Evidence from genome-wide and candidate gene association studies, familial aggregation and linkage analyses demonstrate the genetic contribution to fibromyalgia (FM) disease. This study aimed to identify genetic biomarkers of FM and its related comorbid disorders, by exploring 41 polymorphisms potentially involved in FM pathogenesis in families with at least one patient with FM. METHODS: Core symptoms were assessed, and blood samples collected from 556 patients with FM and 395 healthy relatives. For the genetic study, a final sample of 401 FM patients and 232 healthy controls was selected, discarding patients with concomitant pathologies and controls with chronic pain. A family-based approach using DFAM test (Plink) and SNPs (single nucleotide polymorphisms) combination analyses to compare FM patients vs. controls were first applied. Second, the genotypic distribution of subgroups of FM patients, stratified by severe vs. mild symptoms of pain, depression and sleep impairment, was considered. RESULTS: No evidence of associations with FM per se were detected, using either a family-based approach or SNPs combination analyses. However, considering the subgroups of FM patients, the SNP rs6454674 (CNR1, cannabinoid receptor 1 gene) was found as a potential genetic marker of FM correlated with depression (p<.001). CONCLUSIONS: No significant associations using either the family-based analysis or the SNPs combination tests dissociated FM patients and their healthy relatives. FM patients with and without depression showed a significant difference in the genotypic distribution related to the SNP rs6454674 in the cannabinoid receptor 1 gene (CNR1) indicating that FM is not a homogenous disorder.

Corneoconjunctival transposition for the treatment of deep stromal to full-thickness corneal defects in dogs: A multicentric retrospective study of 100 cases (2012-2018).

OBJECTIVE: To describe and evaluate the use of corneoconjunctival transposition (CTT) as a surgical treatment for canine deep stromal ulcers, descemetoceles, and full-thickness corneal defects and to determine its efficacy in preserving corneal graft transparency and vision. ANIMALS STUDIED: One hundred client-owned dogs with deep stromal ulcers, descemetoceles, or full-thickness corneal defects. METHODS: Medical records of canine patients that underwent CCT, from 2012 to 2018, were reviewed. Only, patients with preoperative positive consensual pupillary light and dazzle reflexes were included. RESULTS: There were 59 males and 41 females, from 0.3 to 17 years. Brachycephalic breeds were overrepresented (65%). All patients were unilaterally affected, with 16 deep stromal ulcers, 33 descemetoceles, and 51 corneal perforations, of a median (range) size of 4 (2-8) mm. The central cornea was affected in the majority of cases (57%), and euryblepharon and keratoconjunctivitis sicca were the most common concurrent ocular diseases (42% and 40%, respectively). The graft was most frequently harvested from dorsal (67%), and 9/0 absorbable suture material was used. Intraoperative and postoperative complications were seen in 7 and 21 cases, respectively. Mean follow-up time was 107.8 days. Vision was preserved in 96% patients, with 62% showing faint to mild opacification. Among the statistically analyzed variables, euryblepharon and pigmentary keratitis were found to be significantly associated with greater corneal graft opacification (P = .040 and P = .028, respectively). CONCLUSIONS: Corneoconjunctival transposition is an effective surgical treatment for deep stromal, descemetocele, and full-thickness corneal defects in dogs, achieving a highly satisfactory degree of corneal graft transparency and preserving vision.

Brain Electrical Activity Associated With Visual Attention and Reactive Motor Inhibition in Patients With Fibromyalgia.

OBJECTIVE: Fibromyalgia (FM) is a generalized chronic pain condition associated with multiple cognitive impairments, including altered inhibitory processes. Inhibition is a key component of human executive functions and shares neural substrate with pain processing, which may explain the inhibitory deficits in FM. Here, we investigated the integrity of brain inhibitory mechanisms in these patients. METHODS: We recorded the electroencephalographic activity of 27 patients with FM and 27 healthy controls (HCs) (all women) while they performed a reactive motor inhibition task (the stop-signal paradigm). We analyzed task-induced modulations in electrophysiological markers related to inhibition (N2, P3, and midfrontal theta oscillations) and visual attention (posterior alpha oscillations). RESULTS: The FM group performed the task correctly, with no differences relative to HCs at the behavioral level. We did not find any between-group differences in N2 amplitude (F(1,52) = 0.01, p = .93), P3 amplitude (F(1,52) = 3.46; p = .068), or theta power (F(1,52) = 0.05; p = .82). However, modulation of posterior alpha power after presentation of either the go or stop stimuli was lower in patients than in HCs (F(1,52) = 7.98; p = .007). CONCLUSIONS: N2, P3, theta power, and behavioral results indicate that the mechanisms of motor inhibition are sufficiently preserved to enable correct performance of the stop-signal task in patients with FM. Nevertheless, the lower modulation of alpha suggests greater difficulty in mobilizing and maintaining visual attentional resources, a result that may explain the cognitive dysfunction observed in FM.

Cryopreservation (-20°C) of equine corneoscleral tissue: Microbiological, histological, and ultrastructural study.

OBJECTIVE: To evaluate microbiological, histological, and ultrastructural characteristics of short-term cryopreserved (STC) equine corneoscleral tissue (<1 year), and to compare it with long-term cryopreserved (LTC) tissue (>7 years). ANIMALS STUDIED: Thirty-four healthy equine globes. PROCEDURE: After a decontamination protocol, globes were enucleated and stored at -20°C in broad-spectrum antibiotics. Corneoscleral tissue was evaluated at different storage periods: 1 month-1 year (20 eyes) and 7-9 years (12 eyes). Two eyes were used as controls. Microbiologic study included direct (blood, McConkey, and Sabouraud agars) and enrichment (brain-heart infusion broth) cultures. Cryopreservation artifacts were evaluated by hematoxylin-eosin. Corneoscleral collagen organization and number of normal and dead keratocytes were established by transmission electron microscopy. RESULTS: All microbiologic direct cultures were negative. Enrichment cultures were positive in 12.5% of corneal and 59.4% of scleral tissues (pcornea  = 0.136; psclera  = 1.000). Cryopreservation artifacts were most commonly observed in LTC tissues (P = 0.002). Normal keratocytes were predominant in STC corneas (STC 60% and LTC 0%) and apoptotic ones in LTC (STC 40% and LTC 90%), whereas necrotic keratocytes were only seen in LTC (LTC 10%) (P = 0.001). No structural differences were detected in collagen organization between STC and LTC (pcornea  = 1.000; psclera  = 0.703). CONCLUSIONS: Cryopreservation of equine corneoscleral tissue did not yield direct bacterial contamination. Apoptosis is the main cause of death of cryopreserved equine keratocytes. Based on the lack of significant structural differences between STC and LTC samples, these cryopreserved tissues could potentially be used for tectonic support for at least 9 years without structural or microbiological impediment.

Endothelial cell density and characterization of corneal endothelial cells in the Tawny Owl (Strix aluco) using specular microscopy.

OBJECTIVE: To determine endothelial cell density (ECD) and morphology and morphometry of corneal endothelial cells in the tawny owl (Strix aluco), as well as to report the effects of aging on these parameters. ANIMAL STUDY AND PROCEDURES: Twenty tawny owls were included in the study and classified into 2 groups according to their age: fledglings (<1 year old) and adults (>1 year old). Central corneal endothelium was studied by means of noncontact specular microscopy (Specular Microscope SP-2000P; Topcon, Tokyo, Japan), and results for ECD (cells/mm2 ), mean cell area ((MCA (µm2 )), polymegathism (CV), and pleomorphism (% hexagonal cells) were obtained. Results are described by median, interquartile range (25th, 75th percentiles), and absolute range for ECD, MCA, pleomorphism, and polymegathism. In addition, inferential analyses by Mann-Whitney U test were also performed. A two-tailed Type I error of 5% was established. RESULTS: Results in fledglings were as follows: ECD = 2864 cells/mm2 , MCA = 348 µm2 , % hexagonal cells = 72.75%, and CV = 21. Results in adults were as follows: ECD = 2602 cells/mm2 , MCA = 384 µm2 , % hexagonal cells = 78.83%, and CV = 16. No significant differences in ECD and MCA were seen between the groups (P > .05), although there were significant differences in % hexagonal cells and CV (P < .05). CONCLUSIONS: Tawny owls present a uniform endothelium in cell size and shape, although ECD and MCA differ greatly from other bird species. Differences in ECD and MCA could not be found between fledglings and adults probably because of the youth of adult specimens, although there were differences in pleomorphism and polymegathism.

A multicenter retrospective study on cryopreserved amniotic membrane transplantation for the treatment of complicated corneal ulcers in the dog.

OBJECTIVE: To describe the surgical technique and outcome of amniotic membrane transplantation (AMT) for treatment of complicated corneal ulcers in dogs. ANIMALS STUDIED: One hundred and eleven dogs. PROCEDURE: Medical records (2010-2017) were reviewed to identify canine patients that underwent AMT for the treatment of complicated corneal ulcers. Quantitative and ordinal variables were analyzed using Mann-Whitney U test and Fisher's Exact test. RESULTS: Brachycephalic breeds were overrepresented (74/111; 66.6%). Three dogs were bilaterally affected. Fifty-one had melting ulcers (51/114; 44.7%), 33 stromal ulcers (33/63; 52.4%), 17 descemetoceles (17/63; 27%), and 13 perforations (13/63;20.6%). Mean defect size was 6.2 mm (2-18 mm). Human (32/114; 28.1%) or bovine (82/114; 71.9%) (AmnioVet(R)) cryopreserved amniotic membranes were used. Monolayer (31/114; 27.2%), bilayer (44/114; 38.6%) or multilayer technique (39/114; 34.2%) was performed, being anchored to the limbus (52/114; 45.6%), to the defect (48/114; 42.1%) or both (14/114; 12.3%). Mean epithelial healing and follow-up times were 25.6 days (15-45) and 98.7 days (21-400), respectively. Most common postsurgical complications included pigmentation (10/114; 8.8%) and graft failure (5/114; 4.4%). Depth and size of corneal lesions, human amniotic membrane (AM) and concurrent ocular diseases were associated with more complications (P < 0.05). Good cosmetic and visual outcomes were achieved in 99.1% (113/114) and 97.4% (111/114) of cases, respectively. Zero to low graft opacity was seen in 23.9% of the eyes (27/113), mild to moderate in 64.6% (73/113) and complete in 11.5% (13/113). CONCLUSIONS: Cryopreserved AMT is an effective surgical technique for the treatment of complicated corneal ulcers in the dog, with highly satisfactory visual and cosmetic outcomes.

Modified brow suspension technique for the treatment of pseudoptosis due to redundant frontal skin in the dog: a retrospective study of 25 cases.

PURPOSE: To describe the outcome of modified brow suspension technique for the treatment of upper eyelid pseudoptosis in canine patients with redundant frontal skin folds. METHODS: Medical records from 1999 to 2015 were retrospectively reviewed. Only dogs that underwent modified brow suspension for treating bilateral pseudoptosis were included in the study. Recorded data included breed, age, gender, main ophthalmic complaint, ocular findings, suture material used for brow suspension, number of slings placed, postoperative treatment, complications, follow-up time, and outcome. RESULTS: Twenty-five adult dogs of different gender and breed fulfilled the inclusion criteria, with the Shar-Pei being overrepresented (19/25;76%). The main complaint at first presentation was mucopurulent discharge (10/25;40%) and blepharospasm (10/25;40%). Concurrent corneal disease was diagnosed in 47 eyes (47/50;94%). Monofilament polyamide was the most frequently used suture material (46/50;92%). Number of slings placed varied, with one in two eyes (4%), two in 28 eyes (56%), three in 17 eyes (34%), and four in three eyes (6%). Complications were seen in three eyes (6%): suture rupture (one eye; 2%) and skin abscess (two eyes; 4%). Mean follow-up time was 17.6 months (1-84 months), with no cases of fold dermatitis or dermatologic disease observed within this period. Good esthetic and functional outcomes were obtained in 47 eyes at first intervention (97%) and in all patients after re-intervention. CONCLUSIONS: The modified brow suspension technique described offers a surgical alternative for dogs with pseudoptosis secondary to redundant frontal skin folds that potentially reduces anesthetic time and facial changes.

Effects of age and breed on corneal thickness, density, and morphology of corneal endothelial cells in enucleated sheep eyes.

OBJECTIVE: The purpose of the study was to determine corneal thickness (CT), endothelial cell density (ECD), morphology, and morphometry in ovine eyes, and to report the effects of age and breed on these parameters. ANIMALS STUDIED AND PROCEDURES: Forty-eight healthy ovine eyes were classified into six groups (eight eyes per group) according to animals' age (lambs and adults) and breed (Manchega, Ripollesa, and Rasa Aragonesa). Endothelial cell density (cells/mm(2) ) and CT (µm) were calculated in the central cornea and four peripheral quadrants by means of specular microscopy. Mean cell area (MCA (µm(2) )), pleomorphism (% hexagonal cells), and polymegethism (CV) were evaluated in the central cornea. Statistics consisted of an ANOVA model and a LSD test for the post hoc analysis. RESULTS: Results in lambs were as follows: ECD = 3532 ± 259, CT = 699 ± 56, MCA = 283 ± 22, % hexagonal cells = 75.64 ± 3.09, and CV = 18 ± 8.4; and in adults: ECD = 2204 ± 261, CT = 804 ± 87, MCA = 473 ± 53, % hexagonal cells = 69.9 ± 3.42, and CV = 18.1 ± 5.2. There were differences (P < 0.001) in all parameters studied between both groups except for the CV (P = 0.962). Regarding the breed, Rasa Aragonesa presented a higher CT (P = 0.001) and CV (P = 0.04) compared to other breeds. Mean values of all groups together showed that central and dorsal areas of the cornea were the thickest, whereas temporal quadrant was the thinnest (P < 0.001). CONCLUSIONS: This study demonstrates that age and breed influence corneal and endothelial parameters in sheep.

Cryopreservation (-20 °C) of feline corneoscleral tissue: histologic, microbiologic, and ultrastructural study.

OBJECTIVE: To evaluate microbiological, histologic, and ultrastructural characteristics of short-term cryopreserved (STC) feline corneoscleral tissue (<1 year) and to compare it with long-term cryopreserved (LTC) tissue (>7 years). ANIMALS STUDIED: Twenty healthy feline globes were obtained from 2003 to 2013. PROCEDURE: After a decontamination protocol, globes were enucleated and stored at -20 °C in broad-spectrum antibiotics. Corneoscleral tissue was evaluated at different storage periods: <1 year (10 eyes) and >7 years (8 eyes). Two eyes were used as controls. Microbiologic study included direct (blood, McConkey, and Sabouraud agars) and enrichment (brain-heart infusion broth) cultures. Cryopreservation artifacts were evaluated by hematoxylin-eosin. Corneoscleral collagen organization and number of normal and dead keratocytes were established by transmission electron microscopy. RESULTS: Although microbiologic cultures were positive only in STC [direct (20.8%); enrichment (37.5%)], significant differences between periods were only found in enrichment cultures (P = 0.006). Cryopreservation artifacts were most commonly observed in LTC tissues (P < 0.001). Normal keratocytes were predominant in STC corneas (STC 58.3%, LTC 12.5%) and apoptotic ones in LTC (STC 41.7%, LTC 75%), whereas necrotic keratocytes were only seen in LTC (LTC 12.5%) (P = 0.046). No structural differences were detected in collagen organization between STC and LTC (Pcornea = 0.147; Psclera = 0.362). CONCLUSIONS: Cryopreservation of feline corneoscleral tissue seems to reduce bacterial contamination over time. Apoptosis is the main cause of death of cryopreserved feline keratocytes. Based on the lack of significant structural differences between STC and LTC samples, these cryopreserved tissues could potentially be used for tectonic support for at least 10 years without structural or microbiological impediment.

The polymorphism Val158Met in the COMT gene: disrupted dopamine system in fibromyalgia patients?

ABSTRACT: The single-nucleotide polymorphism (SNP) rs4680 in the catechol-O-methyltransferase gene ( COMT ) is a missense variant (Val158Met) associated with altered activity of the COMT enzyme and suggested as a predictive feature for developing some chronic pain conditions. However, there are controversial results on its role in fibromyalgia (FM). Here, the SNP Val158Met was analyzed in 294 FM patients (without comorbidities) and 209 healthy controls (without chronic pain). The concurrent impact of Val158Met genotypes and FM comorbid disorders (depression and sleep impairment) on FM risk were tested. In addition, the genotypic distribution of FM patients in relation to pain intensity was evaluated. The G allele (Val) resulted in being more represented in the FM group (57.8%) compared with the control group (48.8%; P = 0.037). Logistic regression highlighted that having the G/G (Val/Val) homozygous genotype was associated with 2 times higher risk of having FM compared with the A/A (Met/Met) carriers ( P = 0.038), whereas depression and sleep impairment increased FM risk by 12 and 8 times, respectively ( P < 0.001). However, considering only the FM patient group, the A/A homozygous genotype was significantly associated with severe pain intensity ( P = 0.007). This study highlighted associations between the SNP Val158Met and both FM and pain intensity, suggesting a link between dopaminergic dysfunction and vulnerability to chronic pain. Further studies should explore this SNP in FM patients in conjunction with COMT enzymatic activity and other symptoms connected with the dopaminergic system such as depression or sleep impairment.

The Armadillo as a Model for Leprosy Nerve Function Impairment: Preventative and Therapeutic Interventions.

Mycobacterium leprae infection of peripheral nerves and the subsequent nerve function impairment (NFI), especially in response to reactional episodes, are hallmarks of leprosy. Improved treatments for M. leprae-induced nerve injury are needed, as most if not all of the disability and stigma associated with leprosy arises from the direct or indirect effects of NFI. Nine-banded armadillos (Dasypus novemcinctus), like humans, exhibit the full clinical spectrum of leprosy and extensive involvement of the peripheral nerves. In this study, state-of-the-art technology was used to compare nerve function between uninfected and M. leprae-infected armadillos. Motor nerve conduction velocity (MNCV) and compound muscle action potential (cMAP), which measure changes in the rate of impulse conduction velocity and amplitude, revealed a progression of impairment that was directly correlated with the duration of M. leprae infection and enabled development of an objective nerve impairment scoring system. Ultrasonography accompanied by color Doppler imaging detected enlargement of the M. leprae-infected nerves and increased vascularity, possibly due to inflammation. Assessment of epidermal nerve fiber density (ENFD), which shows a length-dependent innervation in armadillos that is similar to humans, identified small fiber degeneration early after M. leprae infection. Staining for neuromuscular junction (NMJ) integrity, which is an indicator of signal transduction efficiency into skeletal muscle, discerned a markedly lower number and structural integrity of NMJ in M. leprae-infected armadillo footpads. These tools for assessing nerve injury were used to monitor the effects of intervention therapy. Two potential neuro-protective drugs, ethoxyquin (EQ) and 4-aminopyridine (4-AP), were tested for their ability to ameliorate peripheral nerve injury in M. leprae-infected armadillos. 4-AP treatment improved MNCV, cMAP, and EFND compared to untreated animals, while EQ had less effect. These results support the armadillo as a model for M. leprae-induced peripheral nerve injury that can provide insights toward the understanding of NFI progression and contribute to the preclinical investigation of the safety and efficacy of neuro-preventive and neuro-therapeutic interventions for leprosy.

Quality of life in patients with fibromyalgia: Contributions of disease symptoms, lifestyle and multi-medication.

Fibromyalgia (FM) is a disease characterized by the presence of chronic and widespread musculoskeletal pain, which causes a high negative impact on the quality of life (QoL). Although there are many studies about the QoL of patients with FM, it is unknown which variables have a main influence on it. Therefore, in the present study, we aimed to determine which FM symptoms predict a worse QoL and also to establish whether lifestyle and multi-medication are associated to QoL. We assessed a sample of 134 women with FM using a semi-structured clinical interview to explore lifestyle (diet, exercise, smoking) and medication use, and questionnaires to cover the main symptoms of this disease and QoL (SF-36). We found that the patients with FM had a poor QoL, being "physical pain" and "vitality" the most affected domains. A linear regression analysis showed that depression and anxiety assessed by HADS were the FM symptoms which most significantly predicted QoL, explaining 49% of the variance. Concerning lifestyle/medication influences, we found that multiple drug treatment and smoking also predicted a worse QoL (14%). Moreover, patients who practiced exercise regularly showed better QoL than patients who did not (regardless of the severity of FM). Thus, our results suggest that treatment strategies to improve QoL in FM should be focused on improving psychological distress, promoting regular exercise and reducing smoking and multi-medication. The data highlights the role of positive self-management practices to improve QoL in FM.

Working Memory Performance, Pain and Associated Clinical Variables in Women With Fibromyalgia.

Working memory (WM) is a critical process for cognitive functioning in which fibromyalgia (FM) patients could show cognitive disturbances. Dyscognition in FM has been explained by interference from pain processing, which shares the neural substrates involved in cognition and may capture neural resources required to perform cognitive tasks. However, there is not yet data about how pain is related to WM performance, neither the role that other clinical variables could have. The objectives of this study were (1) to clarify the WM status of patients with FM and its relationship with nociception, and (2) to determine the clinical variables associated to FM that best predict WM performance. To this end, 132 women with FM undertook a neuropsychological assessment of WM functioning (Digit span, Spatial span, ACT tests and a 2-Back task) and a complete clinical assessment (FSQ, FIQ-R, BDI-1A, HADS, PSQI, MFE-30 questionnaires), including determination of pain thresholds and tolerance by pressure algometry. Patients with FM seem to preserve their WM span and ability to maintain and manipulate information online for both visuospatial and verbal domains. However, up to one-third of patients showed impairment in tasks requiring more short-term memory load, divided attention, and information processing ability (measured by the ACT task). Cognitive performance was spuriously related to the level of pain experienced, finding only that pain measures are related to the ACT task. The results of the linear regression analyses suggest that sleep problems and fatigue were the variables that best predicted WM performance in FM patients. Future research should take these variables into account when evaluating dyscognition in FM and should include dynamic measures of pain modulation.

DNA Methylation Changes in Fibromyalgia Suggest the Role of the Immune-Inflammatory Response and Central Sensitization.

Fibromyalgia (FM) has been explained as a result of gene-environment interactions. The present study aims to verify DNA methylation differences in eleven candidate genome regions previously associated to FM, evaluating DNA methylation patterns as potential disease biomarkers. DNA methylation was analyzed through bisulfite sequencing, comparing 42 FM women and their 42 healthy sisters. The associations between the level of methylation in these regions were further explored through a network analysis. Lastly, a logistic regression model investigated the regions potentially associated with FM, when controlling for sociodemographic variables and depressive symptoms. The analysis highlighted significant differences in the GCSAML region methylation between patients and controls. Moreover, seventeen single CpGs, belonging to other genes, were significantly different, however, only one cytosine related to GCSAML survived the correction for multiple comparisons. The network structure of methylation sites was different for each group; GRM2 methylation represented a central node only for FM patients. Logistic regression revealed that depressive symptoms and DNA methylation in the GRM2 region were significantly associated with FM risk. Our study encourages better exploration of GCSAML and GRM2 functions and their possible role in FM affecting immune, inflammatory response, and central sensitization of pain.

DNA methylation changes in genes involved in inflammation and depression in fibromyalgia: a pilot study.

OBJECTIVES: The present pilot study aims to investigate DNA methylation changes of genes related to fibromyalgia (FM) development and its main comorbid symptoms, including sleep impairment, inflammation, depression and other psychiatric disorders. Epigenetic modifications might trigger or perpetuate complex interplay between pain transduction/transmission, central pain processing and experienced stressors in vulnerable individuals. METHODS: We conducted DNA methylation analysis by targeted bisulfite NGS sequencing testing differential methylation in 112 genomic regions from leukocytes of eight women with FM and their eight healthy sisters as controls. RESULTS: Tests for differentially methylated regions and cytosines brought focus on the GRM2 gene, encoding the metabotropic glutamate receptor2. The slightly increased DNA methylation observed in the GRM2 region of FM patients may confirm the involvement of the glutamate pathway in this pathological condition. Logistic regression highlighted the simultaneous association of methylation levels of depression and inflammation-related genes with FM. CONCLUSIONS: Altogether, the results evidence the glutamate pathway involvement in FM and support the idea that a combination of methylated and unmethylated genes could represent a risk factor to FM or its consequence, more than single genes. Further studies on the identified biomarkers could contribute to unravel the causative underlying FM mechanisms, giving reliable directions to research, improving the diagnosis and effective therapies.

Feline leishmaniosis: diagnosis, treatment and outcome in 16 cats.

OBJECTIVES: Leishmaniosis is a vector-borne disease and in European countries is caused by Leishmania infantum. Cats are considered secondary reservoirs of the infection in endemic areas. The objective of this retrospective study is to describe the clinical findings, diagnosis, treatment and outcome of feline leishmaniosis (FeL) in 16 cats in Spain. METHODS: Medical records of cats diagnosed with leishmaniosis were retrospectively reviewed for cases that met the following inclusion criteria: identification of Leishmania organisms and/or DNA on cytological and/or histological specimens and/or a high anti-Leishmania antibody titre, compatible clinical findings and pathological abnormalities. RESULTS: Sixteen cats met the inclusion criteria, all of which were living in areas endemic for canine leishmaniosis. Systemic signs were present in 11 cases (68.8%). The most common clinical signs on presentation included cutaneous lesions in 12 cats (75%), ocular disease in six cats (37.5%) and anorexia in six cats (37.5%). A polyclonal gammopathy was noted in 12 cats (85.7%). Non-regenerative anaemia and renal abnormalities were present in six (37.5%) and five patients (31.3%), respectively. In nine cats (56.3%), immunosuppressive conditions/comorbidities were identified. The diagnosis was made in eight of the cats (50%) by cytology, but a combination of diagnostic tests was needed for definitive diagnosis in the remaining patients. Twelve cats (75%) were treated specifically for leishmaniosis. Five of the 12 cats (41.7%) did not improve with treatment. The median survival time in the group of patients treated specifically for leishmaniosis was 17 months. Median survival of patients treated with concomitant diseases was 13 months vs 41 months in those without, although this was not statistically significant (P = 0.557). CONCLUSIONS AND RELEVANCE: Presentation of FeL appears to be similar to canine leishmaniosis but with some specific features: ulcerative and nodular skin lesions are the predominant cutaneous signs; cats with immunosuppressive conditions or coexisting diseases were more commonly present than typically seen in dogs (mainly feline immunodeficiency virus). A combination of diagnostic tests may be needed for definitive diagnosis.

Serological and molecular detection of infection with Mycobacterium leprae in Brazilian six banded armadillos (Euphractus sexcinctus).

Leprosy was recognized as a zoonotic disease, associated with nine-banded armadillos (Dasypus novemcinctus) in the Southern United States of America in 2011. In addition, there is growing evidence to support a role for armadillos in zoonotic leprosy in South America. The current study evaluated twenty specimens of the six-banded armadillo (Euphractus sexcinctus), collected from rural locations in the state of Rio Grande do Norte (RN), Brazil for evidence of infection with Mycobacterium leprae. Serum was examined using two "in-house" enzyme-linked immunosorbent assays (ELISAs) and via two commercially available (ML flow and NDO-LID®) immunochromatographic lateral flow (LF) tests, for detection of the PGL-I and/or LID-1 antigens of the bacterium. The presence of M. leprae DNA in liver tissue was examined using the multi-copy, M. leprae-specific repetitive element (RLEP), as target in conventional and nested PCR assays. Molecular and anti-PGL-I-ELISA data indicated that 20/20 (100 %) of the armadillos were infected with M. leprae. The corresponding detection levels recorded with the LF tests were 17/20 (85 %) and 16/20 (85 %), for the NDO-LID® and ML flow tests, respectively. Our results indicate that, in common with D. novemcinctus, six banded armadillos (a species hunted and reared as a food-source in some regions of Brazil, including RN), represent a potential reservoir of M. leprae and as such, their role in a possible zoonotic cycle of leprosy within Brazil warrants further investigation.

Fournier's gangrene in a 44-year-old woman: CT scan findings.

OBJECTIVES: To show standard CT findings and their diagnostic usefulness in female patients suffering from Fournier's gangrene. METHOD/RESULT: A woman who had undergone a previous lateral internal sphincterotomy presented to the emergency department with severe pain in the hypogastrium and perianal region; physical examination revealed an induration in the left buttock. CT images showed an abscessed collection in the rectovaginal space and gas in the levator ani muscle, left ischiorectal fossa and the root of the left thigh. CONCLUSIONS: CT scan is considered an excellent diagnostic tool in the management of patients with Fournier's gangrene, as it shows both the origin of the infection and its extent.