Temporomandibular joint injections in dogs with temporomandibular joint pain: 11 cases (2015-2019).

OBJECTIVES: To describe and evaluate the clinical application of temporomandibular joint injections using betamethasone and ropivacaine in German Shepherd dogs suffering from non-odontogenic orofacial pain due to temporomandibular dysplasia and/or osteoarthritis. MATERIALS AND METHODS: Outcomes in dogs presented with clinical signs of non-odontogenic orofacial pain associated to temporomandibular joint dysplasia and/or arthritis and treated with a temporomandibular joint injection were retrospectively-prospectively evaluated. RESULTS: The overall clinical signs free period ranged between 25 to 1579 days, with an average of 461 days. The clinical signs free period for temporomandibular joint osteoarthritis scores 1, 2 and 3 were on average 659 days (180-1579 days), 134 days (42-355 days) and 723 days (25-1377 days), respectively. Similarly the temporomandibular dysplasia scores 1, 2 and 3 were on average 306 days (26-1579 days), 1377 days and 669 days (25-1429 days) respectively. Those dogs in which only one side was injected the clinical signs free period average was 639 days (25-1578 days), compared with dogs in which both temporomandibular joints were injected showing a clinical signs free period average of 378 days (42-1377 days). CLINICAL SIGNIFICANCE: The temporomandibular joint injection technique proved to be feasible with a decent outcome in dogs suffering from non-odontogenic orofacial pain associated with temporomandibular joint osteoarthritis and/or dysplasia. Further randomised studies are required to confirm the effectiveness of this intervention.

100% peer review in radiation oncology: is it feasible?

PURPOSE: Peer review has been proposed as a strategy to ensure patient safety and plan quality in radiation oncology. Despite its potential benefits, barriers commonly exist to its optimal implementation in daily clinical routine. Our purpose is to analyze peer-review process at our institution. METHODS AND MATERIALS: Based on our group peer-review process, we quantified the rate of plan changes, time and resources needed for this process. Prospectively, data on cases presented at our institutional peer-review conference attended by physicians, resident physicians and physicists were collected. Items such as time to present per case, type of patient (adult or pediatric), treatment intent, dose, aimed technique, disease location and receipt of previous radiation were gathered. Cases were then analyzed to determine the rate of major change, minor change and plan rejection after presentation as well as the median time per session. RESULTS: Over a period of 4 weeks, 148 cases were reviewed. Median of attendants was six physicians, three in-training-physicians and one physicist. Median time per session was 38 (4-72) minutes. 59.5% of cases presented in 1-4 min, 32.4% in 5-9 min and 8.1% in ≥ 10 min. 79.1% of cases were accepted without changes, 11.5% with minor changes, 6% with major changes and 3.4% were rejected with indication of new presentation. Most frequent reason of change was contouring corrections (53.8%) followed by dose or fractionation (26.9%). CONCLUSION: Everyday group consensus peer review is an efficient manner to recollect clinical and technical data of cases presented to ensure quality radiation care before initiation of treatment as well as ensuring department quality in a feedback team environment. This model is feasible within the normal operation of every radiation oncology Department.

Identification of putative miRNA biomarkers in early rheumatoid arthritis by genome-wide microarray profiling: A pilot study.

Despite the existing research, the etiology of rheumatoid arthritis (RA), an autoimmune disease remains poorly understood with early and accurate diagnosis difficult to achieve. MicroRNAs (miRNAs) play an important role in biological processes as modulators of transcription and translation. Previous studies have demonstrated a downregulation of several genes in early RA stages and in addition, miRNAs may serve as early biomarkers of subclinical changes in early RA. When comparing the four groups (ANOVA P < 0.01, fold change > 4), we found 253 differentially expressed miRNAs. Of these, 97 miRNAs were identified as overexpressed in early rheumatoid arthritis. The validation of miRNA microarray expression was performed in a set by RT-qPCR and showed strong agreement with microarray expression data. The putative targets of overexpressed microRNAs in early RA were significantly enriched in apoptosis, tolerance loss and Wnt pathways. Moreover, ROC analysis showed values of AUC 0.76 and P < 0.05 for miR 361-5p, identifying this miRNA as a potential biomarker of disease. We identified specific microRNAs associated with early rheumatoid arthritis and proposed them as early biomarkers of disease. Our results provide novel insight into immune disease physiopathology and describe unreported microRNAs in RA with potential for clinical use.

Biogeographical origin and timing of the founder ichthyosis TGM1 c.1187G > A mutation in an isolated Ecuadorian population.

An unusually high frequency of the lamellar ichthyosis TGM1 mutation, c.1187G > A, has been observed in the Ecuadorian province of Manabí. Recently, the same mutation has been detected in a Galician patient (Northwest of Spain). By analyzing patterns of genetic variation around this mutation in Ecuadorian patients and population matched controls, we were able to estimate the age of c.1187G > A and the time to their most recent common ancestor (TMRCA) of c.1187G > A Ecuadorian carriers. While the estimated mutation age is 41 generations ago (~1,025 years ago [ya]), the TMRCA of Ecuadorian c.1187G > A carrier haplotypes dates to just 17 generations (~425 ya). Probabilistic-based inferences of local ancestry allowed us to infer a most likely European origin of a few (16% to 30%) Ecuadorian haplotypes carrying this mutation. In addition, inferences on demographic historical changes based on c.1187G > A Ecuadorian carrier haplotypes estimated an exponential population growth starting ~20 generations, compatible with a recent founder effect occurring in Manabí. Two main hypotheses can be considered for the origin of c.1187G > A: (i) the mutation could have arisen in Spain >1,000 ya (being Galicia the possible homeland) and then carried to Ecuador by Spaniards in colonial times ~400 ya, and (ii) two independent mutational events originated this mutation in Ecuador and Galicia. The geographic and cultural characteristics of Manabí could have favored a founder effect that explains the high prevalence of TGM1 c.1187G > A in this region.

Validation of the prostate health index in a predictive model of prostate cancer. / Validación del índice de salud prostática en un modelo predictivo de cáncer de próstata.

OBJECTIVES: To validate and analyse the clinical usefulness of a predictive model of prostate cancer that incorporates the biomarker «[-2] pro prostate-specific antigen¼ using the prostate health index (PHI) in decision making for performing prostate biopsies. MATERIAL AND METHODS: We isolated serum from 197 men with an indication for prostate biopsy to determine the total prostate-specific antigen (tPSA), the free PSA fraction (fPSA) and the [-2] proPSA (p2PSA). The PHI was calculated as p2PSA/fPSA×√tPSA. We created 2 predictive models that incorporated clinical variables along with tPSA or PHI. The performance of PHI was assessed with a discriminant analysis using receiver operating characteristic curves, internal calibration and decision curves. RESULTS: The areas under the curve for the tPSA and PHI models were 0.71 and 0.85, respectively. The PHI model showed a better ability to discriminate and better calibration for predicting prostate cancer but not for predicting a Gleason score in the biopsy ≥7. The decision curves showed a greater net benefit with the PHI model for diagnosing prostate cancer when the probability threshold was 15-35% and greater savings (20%) in the number of biopsies. CONCLUSIONS: The incorporation of p2PSA through PHI in predictive models of prostate cancer improves the accuracy of the risk stratification and helps in the decision-making process for performing prostate biopsies.

Autologous stem cell transplantation (ASCT) in patients with mantle cell lymphoma: a retrospective study of the Spanish lymphoma group (GELTAMO).

Guidelines recommend autologous stem cell transplantation (ASCT) consolidation in first complete or partial response after regimens including rituximab (R) and high-dose AraC (HDAC), but its use beyond that response is questioned. We present a retrospective analysis of 268 patients with MCL who received ASCT. With a median follow-up for survival patients of 54 months, progression-free survival and overall survival for the whole series were 38 and 74 months, respectively, and for patients transplanted in first CR 49 and 97 months, respectively. Patients without CR before transplant were analyzed separately, those who achieved CR after transplantation had better PFS (48 vs 0.03 months, p < 0.001) and OS (92 vs 16 months, p < 0.001) than the remaining. In univariate analysis, first CR at transplant (p = 0.01) and prior rituximab (p = 0.02) were the variables associated with PFS. For OS, the same variables resulted significant (p = 0.03 and p < 0.001, respectively). In multivariate analysis, only the status at transplant (first CR) remained significant. This retrospective study concludes that ASCT consolidation in first CR induces high survival rates. In other stages of disease, the need of ASCT as consolidation may be questioned.

Increased expression of microRNA-155 in peripheral blood mononuclear cells from psoriasis patients is related to disease activity.

BACKGROUND: MicroRNAs (miRNAs) gene expression regulators are altered in psoriasis suggesting their role in the pathogenesis. OBJECTIVE: To study expression changes of inflammation and toll-like receptor (TLR)-related miRNAs, miRNA-155, let-7i, miRNA-21, miRNA-146a and miRNA-223 in peripheral mononuclear cells (PBMCs) and miRNA-21, miRNA-146a and miRNA-223 in plasma, from chronic plaque-type psoriasis patients who were treatment-naive or had undergone a washout period (n = 11). MiRNAs were evaluated at baseline and after 11 (9-12) months [median (25th-75th percentile range)] of methotrexate (MTX) or topical (betamethasone plus calcipotriene) treatment. METHODS: MiRNA expression was analysed with quantitative real-time reverse transcription-polymerase chain reaction. Matched controls were studied. RESULTS: Psoriasis patients presented, at baseline, increased expression of miRNA-155, let-7i, miRNA-146a, miRNA-21 and miRNA-223 in PBMCs, plus miRNA-21, miRNA-146a and miRNA-223 in plasma. Receiver-operator characteristic (ROC) curve analysis and area under the curve (AUC) showed that expression of these miRNAs have the potential to distinguish between psoriasis and controls. At baseline, miRNA-155 expression in PBMCs correlated with Psoriasis Area Severity Index (PASI) [12 (8-14)] (Spearman r: 0.7140, P < 0.05) suggesting a role in psoriasis. After MTX or topical treatment, reduction in PASI was observed [87.5% (75-100)]; miRNA-155 expression in PBMCs decreased; plasma miRNA-21, miRNA-146a and miRNA-223 were down-regulated. ROC analysis showed that miRNA-155 expression in PBMCs from psoriasis patients have the potential to distinguish between patients' samples at baseline and after treatment (AUC: 0.942, sensitivity: 0.91; specificity: 0.91 values; maximum likelihood ratio =10). After treatment, miRNA-146a expression in PBMCs increased; miRNA-155/miRNA-146a ratio decreased, suggestive of a regulatory feedback; let-7i expression decreased; miRNA-21 and miRNA-223 remained elevated. CONCLUSION: In this exploratory study, psoriasis patients presented increased expression of miRNA-155 in PBMCs that correlated with PASI and decreased with disease remission. MiRNA-21, miRNA-146a and miRNA-223 in PBMCs and plasma were increased at baseline and differentially modulated, underscoring different roles of TLR-related miRNAs in psoriasis.

Implementation of a National Priority Allocation System for Hypersensitized Patients in Spain, Based on Virtual Crossmatch: Initial Results.

Access to kidney transplantation for patients with high levels of antibodies against HLA is a major challenge. This issue makes it difficult to detect compatible donors for those patients in a certain geographical area. Consequently, hypersensitized patients remain on the waiting list for long periods and their quality of life deteriorates. Our purpose was to increase access to transplantation for highly sensitized patients by developing a national priority allocation system based on virtual crossmatch. Between June 15, 2015, and May 15, 2016, 675 patients on the kidney transplant waiting list with calculated panel-reactive antibodies ≥98% and undergoing dialysis for at least 12 months were included in the study; 86.1% of the patients had previously received at least one transplant. Solid-phase immunoassays were used to identify class I and II HLA antibodies in all patients. Participating hospitals assigned to the program one of the kidneys of every identified brain-dead real donor between 18 and 70 years old. Survival data were collected for the recipients transplanted between June 15, 2015, and December 31, 2015. In all, 475 (290 male and 185 female) brain-dead donors were assigned to the program. Virtual crossmatch was negative for 191 (41%) donors, 149 offers were accepted, and 102 (21.8%) kidneys were transplanted. At the end of the study, patient and graft survival were both 93.4%. The implementation of a national prioritization system based on virtual crossmatch increased access to transplantation for highly sensitized patients, with excellent results in terms of patient and graft survival.

Antibody-Mediated Rejection in Kidney Transplantation Without Evidence of Anti-HLA Antibodies?

INTRODUCTION: The definition of antibody-mediated rejection (AMR) is based on serologic (presence and/or development of donor-specific anti-HLA antibodies [DSAs]) and histologic (C4d deposition and endothelial damage) criteria. However, several cases of AMR have been described without C4d deposition, and other cases of histologic AMR without DSAs, which could be driven by other non-HLA alloantibodies such as anti-MICA or anti-angiotensin II type I receptor (AT1R). Here we studied clinical and histologic humoral rejection in kidney transplant recipients without evidence of anti-HLA antibodies. MATERIALS AND METHODS: Fifteen kidney transplant recipients with AMR defined as C4d+ and/or histologic g+ptc without anti-HLA antibodies in screening test were studied. Sera at the moment of biopsy and 2 months earlier were studied for anti-HLA antibodies by Luminex, in neat, diluted 1/160, and sera after treatment with dithiothreitol (DTT) and confirmed by single-antigen test. The anti-AT1R was measured by enzyme-linked immunosorbent assay. RESULTS: A lack of anti-HLA and MICA antibodies was confirmed after anti-HLA screening test in all conditions (neat, diluted, and DTT-treated) and de novo development of AT1R antibodies was ruled out. Nevertheless, after single-antigen test, 3 patients were identified with a weak reaction against class I antigen and another 4 patients against class II antigen. Due to the lack of locus-C typing in the donors, the DSA assignment cannot be confirmed, whereas anti-HLA class II antigens were DSA. CONCLUSIONS: A low sensitivity in the screening of anti-HLA antibody testing was observed. Our results suggest performing single-antigen test in seronegative patients with clinical humoral rejection after screening to confirm the presence of DSA.

Detection of new pathogenic mutations in patients with congenital haemolytic anaemia using next-generation sequencing.

INTRODUCTION: Congenital haemolytic anaemia (CHA) refers to a group of genetically heterogeneous disorders, mainly caused by changes in genes encoding globin chains, cytoskeletal proteins and red cell enzymes, in which accurate diagnosis can be challenging with conventional techniques. METHODS: To set-up a comprehensive assay for detecting mutations that could improve aetiological diagnosis, we designed a custom panel for sequencing coding regions from 40 genes known to be involved in the pathogenesis of CHA, using the Ion Torrent™ (Thermo Fisher Scientific, S.L. Waltham, MA, USA) Personal Genome Machine (PGM) Sequencer. A control group of 16 samples with previously known mutations and a test group of 10 patients with unknown mutations were included for assay validation and application, respectively. RESULTS: In the test group, we identified pathogenic mutations in all cases: four patients had novel mutations in genes related to membrane defects (SPTB, ANK1, SLC4A1 and EPB41), four were homozygous or compound heterozygous for mutations in genes related to enzyme deficiencies (GPI, TPI1 and GSS), one had a mutation in the HBB gene and another presented a homozygous mutation in the ADAMTS13 gene. CONCLUSIONS: Ion PGM sequencing with our custom panel is a highly efficient way to detect mutations causing haemolytic anaemia, including new variations. It is a high-throughput detection method that is ready for application in clinical laboratories.

Antidiabetic and antioxidant activity of Stevia rebaudiana extracts (Var. Morita) and their incorporation into a potential functional bread.

In this study a new wheat bread was designed whose sugars were replaced with S. rebaudiana Bertoni aqueous extract. The impact of the S. rebaudiana Bertoni aqueous extract on nutritional and sensory quality, its ability to reduce sugar intake and its antioxidant properties were investigated. Functional bread with 50 % of sugars replaced with S. rebaudiana extract was compared with traditional wheat bread. The extract demonstrated alpha amylase (IC50 = 198.40 µg/mL) glucosidase (596.77 µg/mL) inhibition. The radical scavenging activity exhibited an IC50 value of 335.94 mg/mL. In comparison with the control, the bread with stevia extract was softer and had lower microbial growth during the shelf-life study. The sensory test showed that the substitution of 50 % stevia extract was more acceptable when comparing with all quality characteristics. Regarding the nutritional contribution, the content of dietary fiber and digestible carbohydrates in the bread with stevia extract was higher and lower respectively, so caloric intake was significantly reduced. The results showed that the biological properties of S. rebaudiana extract were retained after the bread making process and that the proposed bread is suitable as functional food in human nutrition.

Frequencies of circulating B-cell subpopulations before kidney transplantation identify patients at risk of acute rejection.

The response mediated by B lymphocytes has a crucial impact on kidney transplantation due to the role of anti-human leukocyte antigen antibodies in rejection and the contradictory observation of high B-lymphocyte numbers in tolerant kidney transplant recipients. The basis of the contradiction could lay in the different function of B-cell subsets depending on their degree of differentiation. We ought to measure circulating B-lymphocyte percentages in patients with end-stage renal disease before kidney transplantation to identify those with a high risk of acute rejection. Eighty patients on the waiting list for kidney transplantation followed up in our center were recruited from 2010, and samples were taken just before kidney transplantation. Eleven of 80 patients presented an episode of acute rejection (13.75%) and had an increased frequency of switched (SW) B cells compared with the rejection-free group (median [interquartile range] 24.5% [18.6% to 39.6%] vs 15.1 [8.45% to 23.4%]; P = .025). Subsequently, the frequency of SW B cells was assessed as a predicting factor of acute rejection. A value higher than 18.4% predicted patients at risk of suffering an acute rejection episode with a sensitivity of 81.8% and a specificity of 60.9% and an area under the curve of 71.2%. Moreover, a decrease in naïve B-cell subsets was related to patients at risk of acute rejection. The percentage of circulating B-cell subsets before kidney transplantation could be used as biomarker of risk to suffer acute rejection. These unicenter data must be validated in multicenter studies.

Number of antihypertensive drugs at 1 year after kidney transplantation.

High blood pressure (BP) affects up to 90% of kidney transplant recipients and is associated with lower patient and graft survival rates. Kidney Disease/Improving Global Outcomes (KDIGO) guidelines suggest maintaining BP at lower than 130/80 mm Hg. Multidrug therapy is usually required for the control of BP in this population. Our aim was to analyze the number of antihypertensive drugs used in our kidney transplantation population at 1 year after transplantation and their influence on graft and patient outcome. We included 411 deceased-donor kidney transplantation cases; data were obtained from a prospectively maintained institutional database. BP was measured at the outpatient clinic. Approximately 97 patients were not under antihypertensive therapy, whereas 130, 119, 52, and 13 received 1, 2, 3, or 4 antihypertensive drugs, respectively. The number of antihypertensive drugs was significantly related to lower patient survival rates independently of a previous diagnosis of hypertension and diabetes, recipient age and sex and renal function at 1-year. After multivariate linear regression analysis high body mass index, male gender of recipients, donor hypertension, previous acute rejection, and cyclosporine therapy were risk factors independently related to a higher number of antihypertensive drugs. To conclude, the number of antihypertensive drugs is an objective and easy-to-measure marker related to lower patient survival rates. Recipient body mass index, type of calcineurin inhibitor, and acute rejection are modifiable risk factors whose control can help to reduce the number of antihypertensive drugs needed to treat high BP in the kidney transplantation population.

A rat knockout model implicates TRPC4 in visceral pain sensation.

Acute and chronic pain resulting from injury, surgery, or disease afflicts >100 million Americans each year, having a severe impact on mood, mental health, and quality of life. The lack of structural and functional information for most ion channels, many of which play key roles in the detection and transmission of noxious stimuli, means that there remain unidentified therapeutic targets for pain management. This study focuses on the transient receptor potential canonical subfamily 4 (TRPC4) ion channel, which is involved in the tissue-specific and stimulus-dependent regulation of intracellular Ca²âº signaling. Rats with a transposon-mediated TRPC4-knockout mutation displayed tolerance to visceral pain induced by colonic mustard oil (MO) exposure, but not somatic or neuropathic pain stimuli. Moreover, wild-type rats treated with a selective TRPC4 antagonist (ML-204) prior to MO exposure mimicked the behavioral responses observed in TRPC4-knockout rats. Significantly, ML-204 inhibited visceral pain-related behavior in a dose-dependent manner without noticeable adverse effects. These data provide evidence that TRPC4 is required for detection and/or transmission of colonic MO visceral pain sensation. In the future, inhibitors of TRPC4 signaling may provide a highly promising path for the development of first-in-class therapeutics for this visceral pain, which may have fewer side effects and less addictive potential than opioid derivatives.

Disminución de los niveles plasmáticos de clusterina en pacientes con psoriasis / Decreased Plasma Levels of Clusterin in Patients With Psoriasis

Introducción y objetivos: La psoriasis es una enfermedad inflamatoria crónica que se ha asociado a un incremento del riesgo cardiovascular. La clusterina (apolipoproteína J) es un componente de las lipoproteínas de alta densidad (HDL) y tiene un papel protector de la ateroesclerosis. El objetivo del estudio ha sido evaluar la clusterina y el factor inhibitorio de la migración del macrófago (MIF) plasmáticos en pacientes con psoriasis grave comparando grupos de pacientes con distintos riesgos cardiovasculares asociados. Material y métodos: Se estudiaron 21 pacientes con psoriasis grave (Psoriasis Area Severity Index [PASI] y Body Surface Area [BSA] > 10) y 11 controles sin enfermedad dermatológica. Se evaluaron los factores de riesgo cardiovascular según criterios del síndrome metabólico del Adult Treatment Panel III (ATP- III ) y la ateromatosis carotídea subclínica mediante ecografía doppler de carótidas. La clusterina y MIF plasmáticos se midieron mediante Enzyme-Linked Immuno Sorbent Assay (ELISA). Resultados: El 47% de los pacientes con psoriasis presentaba criterios de síndrome metabólico y el 33% presentó placa de ateroma carotídea. Se observó una disminución significativa de la clusterina plasmática (μg/ml) en pacientes con psoriasis respecto a controles (81,39 ± 27,30; n = 21, versus 117 ± 21,6, n = 11; p = 0,0017). El MIF plasmático (ng/ml) estaba aumentado significativamente en los pacientes con psoriasis y placa de ateroma carotídea respecto a los controles (53,22 ± 29,02; n = 6, versus 34,21 ± 9,65; n = 11; p = 0,0394). Conclusiones: La disminución de clusterina en pacientes con psoriasis sugiere una relación con la enfermedad y con la situación inflamatoria sistémica. El aumento de MIF en pacientes parece relacionarse con la presencia de factores de riesgo cardiovascular asociados y placa de ateroma carotídea (AU)

Introduction and objectives: Psoriasis is a chronic inflammatory disease that has been linked to increased cardiovascular risk. The glycoprotein clusterin (apolipoprotein J) is a component of high-density lipoproteins and has a protective role in atherosclerosis. The aim of the present study was to evaluate the plasma levels of clusterin and the proinflammatory cytokine macrophage migration inhibitory factor (MIF) in patients with severe psoriasis, comparing groups of patients with different risks of cardiovascular disease. Material and methods: Twenty-one patients with severe psoriasis (psoriasis area severity index and body surface area > 10) and 11 healthy controls with no dermatologic disease were studied. Cardiovascular risk factors were assessed according to the Adult Treatment Panel (ATP) IIIcriteria. Subclinical carotid atheromatosis was assessed by Doppler ultrasonography of the carotid arteries. Plasma clusterin and MIF levels were measured by enzyme-linked immunosorbent assay. Results: ATP-III criteria for metabolic syndrome were met by 47% of the patients, and 33% had carotid atheromatous plaque. Mean (SD) clusterin plasma levels were significantly lower in patients with psoriasis compared with controls (81.39 [27.30] micreg/mL for the 21 patients vs 117[21.6] g/mL for the 11 controls; P = .0017). MIF plasma levels (ng/ml) were significantly higherin patients with atheromatous plaque compared with controls (53.22 [29.02] for the 6 patientswith plaque vs 34.21 [9.65] for the 11 controls; P = 0.0394).Conclusions: The decreased plasma levels of clusterin in psoriatic patients suggested an association with the disease and might be an indicator of systemic inflammatory activity. Increased levels of MIF appear to be associated with cardiovascular risk factors and carotid atheromatous plaque (AU)