Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature

Worldwide cooperation between laboratories working with classical swine fever virus (CSFV) requires exchange of virus isolates. For this purpose, shipment of CSFV RNA is a safe alternative to the exchange of infectious material. New techniques using desiccation have been developed to store RNA at room temperature and are reported as effective means of preserving RNA integrity. In this study, we evaluated the stability and integrity of dried CSFV RNA stored at room temperature. First, we determined the stability of CSFV RNA covering CSFV genome regions used typically for the detection of viral RNA in diagnostic samples by reverse transcription-polymerase chain reaction (RT-PCR). To this end, different concentrations of in vitro-transcribed RNAs of the 5’-untranslated region and of the NS5B gene were stored as dried RNA at 4, 20, and 37oC for two months. Aliquots were analyzed every week by CSFV-specific quantitative real-time RT-PCR. Neither the RNA concentration nor the storage temperature did affect CSFV RNA yields at any of the time evaluated until the end of the experiment. Furthermore, it was possible to recover infectious CSFV after transfection of SK-6 cells with dried viral RNA stored at room temperature for one week. The full-length E2 of CSFV was amplified from all the recovered viruses, and nucleotide sequence analysis revealed 100% identity with the corresponding sequence obtained from RNA of the original material. These results show that CSFV RNA stored as dried RNA at room temperature is stable, maintaining its integrity for downstream analyses and applications

Deciphering predictive factors for choice of thrombopoietin receptor agonist, treatment free responses, and thrombotic events in immune thrombocytopenia

Malalties hematològiques; Trastorns immunològicsEnfermedades hematológicas; Trastornos inmunológicosHaematological diseases; Immunological disordersVery few data exist on when a particular thrombopoietin-receptor agonist (TPO-RA) is favored in clinical practice for the treatment of patients with immune thrombocytopenia (ITP), about novel risk factors for vascular events (VE) with these drugs, nor about predictive factors for therapy free responses (TFR). We conducted an observational, retrospective, long-term follow-up multicenter study from November 2016 to January 2018 of 121 adult ITP patients initiating TPO-RA between January 2012 to December 2014. Data reflected that a platelet count ≤25 × 109/l at the time when the TPO-RA was initiated was associated with a 2.8 higher probability of receiving romiplostim vs. eltrombopag (P = 0.010). VE on TPO-RA was related to previous neoplasia in patients over 65 years (50% vs. 2.2%, P < 0.001), and to previous splenectomy in younger patients (100% vs. 33%, P = 0.001). Receiving romiplostim as first TPO-RA with no subsequent TPO-RA switching was associated with a 50% likelihood of TFR after 2.9 years of therapy (3.3 years in chronic ITP patients). These real-world data help deciphering some areas of uncertainty, and offer insight into some of the most relevant challenges of ITP which may help clinicians make appropriate treatment decisions in the management of adult ITP patients with TPO-RA

The economic impact of machine perfusion technology in liver transplantation

Introduction: Several clinical studies have demonstrated the safety, feasibility, and efficacy of machine perfusion in liver transplantation, although its economic outcomes are still underexplored. This review aimed to examine the costs related to machine perfusion and its associated outcomes.Methods: Expert opinion of several groups representing different machine perfusion modalities. Critical analysis of the published literature reporting the economic outcomes of the most used techniques of machine perfusion in liver transplantation (normothermic and hypothermic ex situ machine perfusion and in situ normothermic regional perfusion).Results: Machine perfusion costs include disposable components of the perfusion device, perfusate components, personnel and facility fees, and depreciation of the perfusion device or device lease fee. The limited current literature suggests that although this upfront cost varies between perfusion modalities, its use is highly likely to be cost-effective. Optimization of the donor liver utilization rate, local conditions of transplant programs (long waiting list times and higher MELD scores), a decreased rate of complications, changes in logistics, and length of hospital stay are potential cost savings points that must highlight the expected benefits of this intervention. An additional unaccounted factor is that machine perfusion optimizing donor organ utilization allows patients to be transplanted earlier, avoiding clinical deterioration while on the waiting list and the costs associated with hospital admissions and other required procedures.Conclusion: So far, the clinical benefits have guided machine perfusion implementation in liver transplantation. Albeit there is data suggesting the economic benefit of the technique, further investigation of its costs to healthcare systems and society and associated outcomes is needed.</p

Personality in patients with eating disorders depending on the presence/absence of comorbidity with borderline personality disorder / Personalidad en pacientes con trastorno alimentario en función de la presencia/ausencia de comorbilidad con trastorno límite de la personalidad

Abstract  Studies on comorbidity between eating disorders (ED) and personality disorders yield rates of 20-80%, a condition that makes difficult the diagnosis and complicates prognosis, especially regarding their comorbidity with borderline personality disorder (BPD). The objective of this study was to assess whether pathological personality alterations make it possible to distinguish not only between different types of ED, but also in terms of their comorbidity with BPD. Participants included 29 patients with ED, 10 with comorbid ED with BPD (EDc), 27 with BPD and 22 healthy controls, who completed the Millon Multiaxial Clinical Inventory (MCMI-II), and two other measures, one on impulsivity and other aimed at assess symptoms of bulimia nervosa (BN). The ED group was characterized by greater compulsivity, mainly in the subgroup with anorexia nervosa vs. BPD and EDc groups; however, these last two groups shared many features that show emotional instability, although less that the subgroup with BN. These findings support a continuum of severity in terms of compulsivity-impulsivity, with important etiopathogenic, diagnostic and psychotherapeutic implications.Resumen   Los estudios sobre comorbilidad entre los trastornos de la conducta alimentaria (TCA) y los trastornos de la personalidad reportan tasas del 20-80%, condición que dificulta el diagnóstico y ensombrece el pronóstico, especialmente respecto a su comorbilidad con el trastorno límite de la personalidad (TLP). El objetivo de este estudio fue valorar si las alteraciones patológicas de la personalidad permiten distinguir no solo entre distintos tipos de TCA, sino además en función de su comorbilidad con el TLP. Participaron 29 pacientes con TCA, 10 con TCA comórbido con TLP [TCAc], 27 con TLP y 22 controles sanas, quienes completaron el Inventario Clínico Multiaxial de Millon (MCMI-II) y otras dos medidas, una sobre impulsividad y otra dirigida a evaluar síntomas de bulimia nerviosa (BN). El grupo TCA se caracterizó por mayor compulsividad, principalmente en el caso del subgrupo con anorexia nerviosa vs. los grupos TLP y TCAc; no obstante, estos dos últimos compartieron un amplio número de rasgos indicativos de inestabilidad emocional, y también –aunque en menor medida− con el subgrupo con BN. Los hallazgos apoyan un continuum de gravedad en función de la compulsividad-impulsividad, con importantes implicaciones etiopatogénicas, diagnósticas y psicoterapéuticas.

Effects and Mechanisms of Cognitive, Aerobic Exercise, and Combined Training on Cognition, Health, and Brain Outcomes in Physically Inactive Older Adults : The Projecte Moviment Protocol

Altres ajuts: It has also been rewarded with three pre-doctoral fellowships ( FPU014/01460, FI-2016, and FI-2018).Introduction: Age-related health, brain, and cognitive impairment is a great challenge in current society. Cognitive training, aerobic exercise and their combination have been shown to benefit health, brain, cognition and psychological status in healthy older adults. Inconsistent results across studies may be related to several variables. We need to better identify cognitive changes, individual variables that may predict the effect of these interventions, and changes in structural and functional brain outcomes as well as physiological molecular correlates that may be mediating these effects. Projecte Moviment is a multi-domain randomized trial examining the effect of these interventions applied 5 days per week for 3 months compared to a passive control group. The aim of this paper is to describe the sample, procedures and planned analyses. Methods: One hundred and forty healthy physically inactive older adults will be randomly assigned to computerized cognitive training (CCT), aerobic exercise (AE), combined training (COMB), or a control group. The intervention consists of a 3 month home-based program 5 days per week in sessions of 45 min. Data from cognitive, physical, and psychological tests, cardiovascular risk factors, structural and functional brain scans, and blood samples will be obtained before and after the intervention. Results: Effects of the interventions on cognitive outcomes will be described in intention-to-treat and per protocol analyses. We will also analyze potential genetic, demographic, brain, and physiological molecular correlates that may predict the effects of intervention, as well as the association between cognitive effects and changes in these variables using the per protocol sample. Discussion: Projecte Moviment is a multi-domain intervention trial based on prior evidence that aims to understand the effects of CCT, AE, and COMB on cognitive and psychological outcomes compared to a passive control group, and to determine related biological correlates and predictors of the intervention effects. Clinical Trial Registration: www.ClinicalTrials.gov, identifier NCT03123900

Toxoplasma gondii and <i>Trichinella</i> infections in wild boars (<i>Sus scrofa</i>) from Northeastern Patagonia, Argentina

Wild boar (Sus scrofa) was introduced in many countries of the world and is recognized as carrier of many infectious diseases. Wild game meat consumption is recognized as a source of transmission of Toxoplasma gondii and Trichinella spp. The aim of the present study was to evaluate the prevalence of antibodies to T. gondii and Trichinella spp. in free-range wild boars in Northeastern Argentine Patagonia. Between 2014 and 2018, 144 blood samples and 423 muscle samples from 423 carcasses were collected. To detect T. gondii IgG, 144 sera were processed by an immunofluorescent antibody test, and to detect anti-Trichinella IgG, 125 sera and 304 muscle juice samples were processed by ELISA. Detection of first stage larvae in muscle was performed by artificial digestion . A total of 423 wild boars muscle samples were negative to Trichinella spp. by artificial digestion. Antibodies to Trichinella spp. were detected in 2.4% (3/125) of serum samples and in 1.64% (5/304) of meat juice samples. Antibodies to T. gondii infection were detected in 12.5% (18/144) of the serum samples. This is the first study to reveal the presence of antibodies to T. gondii in wild boars from Argentina. The present results suggest that consumption of raw or undercooked wild boar meat could represent a potential source risk for toxoplasmosis in humans and that Trichinella spp. is infrequent and/or that it circulates in low burdens among wild boars in Northeastern Patagonia.Laboratorio de Inmunoparasitologí

Digestibilidad in vitro de rebrote del forraje integral de dos nuevos cultivares de caña de azúcar (Saccharum spp. C97-366 y C99-374).

Se determinó la digestibilidad in vitro de dos nuevos cultivares de caña de azúcar para forraje. Se desarrollaron tres corridas con la técnica de producción de gases con heces vacunas como inóculo. Se estudió la fracción integral de la planta de 6; 8 y 11 meses de edad de rebrote. Se utilizó como testigo el cultivar forrajero My5514 y, para determinarlos parámetros, el modelo de Ørskov y McDonald (1979) modificado por Correa (2004). Se realizaron análisis de varianza simple y se determinaron las diferencias entre medias por la prueba de Tukey para P &lt; 0,05. Los resultados mostraron que los dos nuevos cultivares produjeron volúmenes de gas in vitro similares e, incluso, superiores a los 8 y 11 meses de edad, al cultivar forrajero My5514. A los 8 meses los nuevos cultivares mostraron los mayores valores de producción de gas in vitro. Los valores obtenidos para los parámetros de producción de gas in vitro, velocidad(0,011 h-1-0,033 h-1) y potencial de producción de gas (32,7 ml-52,1 ml) fueron altos; por su parte el lag mostró valores bajos (2,10 h-4,57 h).In vitro Digestibility of Whole Forage Shoots from Two New Sugar Cane Cultivars (Saccharumspp. C97-366 y C99-374) ABSTRACTIn vitro digestibility of two new sugar cane cultivars for forage was determined. Three runs were performed using gas from cattle gases as inoculum. The integral fraction of the plant at 6; 8 and 11 months of reshoot were studied. Forage cultivar My5514 was used as witness, and to determine the parameters, the Ørskov and McDonald (1979) model modified by Correa (2004) was used. Simple variance analyses were made and the differences were determined by the Tukey test for P &lt; 0.05. The results showed that the two new cultivars produced similar in vitro gas volumes to My5514 forage cultivar, and were even higher at 8 and 11 months of age. Eight months later, the new cultivars produced the highest in vitro gas values. The values achieved for the production parameters of in vitro gas, speed (0.011 h-1-0.033 h-1) and potential for gas production (32.7 ml-52.1 ml) were high; on its turn, lag had low values (2.10 h-4.57 h)

Monitoring an Alien Invasion: DNA Barcoding and the Identification of Lionfish and Their Prey on Coral Reefs of the Mexican Caribbean

BACKGROUND: In the Mexican Caribbean, the exotic lionfish Pterois volitans has become a species of great concern because of their predatory habits and rapid expansion onto the Mesoamerican coral reef, the second largest continuous reef system in the world. This is the first report of DNA identification of stomach contents of lionfish using the barcode of life reference database (BOLD). METHODOLOGY/PRINCIPAL FINDINGS: We confirm with barcoding that only Pterois volitans is apparently present in the Mexican Caribbean. We analyzed the stomach contents of 157 specimens of P. volitans from various locations in the region. Based on DNA matches in the Barcode of Life Database (BOLD) and GenBank, we identified fishes from five orders, 14 families, 22 genera and 34 species in the stomach contents. The families with the most species represented were Gobiidae and Apogonidae. Some prey taxa are commercially important species. Seven species were new records for the Mexican Caribbean: Apogon mosavi, Coryphopterus venezuelae, C. thrix, C. tortugae, Lythrypnus minimus, Starksia langi and S. ocellata. DNA matches, as well as the presence of intact lionfish in the stomach contents, indicate some degree of cannibalism, a behavior confirmed in this species by the first time. We obtained 45 distinct crustacean prey sequences, from which only 20 taxa could be identified from the BOLD and GenBank databases. The matches were primarily to Decapoda but only a single taxon could be identified to the species level, Euphausia americana. CONCLUSIONS/SIGNIFICANCE: This technique proved to be an efficient and useful method, especially since prey species could be identified from partially-digested remains. The primary limitation is the lack of comprehensive coverage of potential prey species in the region in the BOLD and GenBank databases, especially among invertebrates

BCL3-rearrangements in B-cell lymphoid neoplasms occur in two breakpoint clusters associated with different diseases

The t(14;19)(q32;q13) often juxtaposes BCL3 with immunoglobulin heavy chain (IGH) resulting in overexpression of the gene. In contrast to other oncogenic translocations, BCL3 rearrangement (BCL3-R) has been associated with a broad spectrum of lymphoid neoplasms. Here we report an integrative whole-genome sequence, transcriptomic, and DNA methylation analysis of 13 lymphoid neoplasms with BCL3-R. The resolution of the breakpoints at single base-pair revealed that they occur in two clusters at 5' (n=9) and 3' (n=4) regions of BCL3 associated with two different biological and clinical entities. Both breakpoints were mediated by aberrant class switch recombination of the IGH locus. However, the 5' breakpoints (upstream) juxtaposed BCL3 next to an IGH enhancer leading to overexpression of the gene whereas the 3' breakpoints (downstream) positioned BCL3 outside the influence of the IGH and were not associated with its expression. Upstream BCL3-R tumors had unmutated IGHV, trisomy 12, and mutated genes frequently seen in chronic lymphocytic leukemia (CLL) but had an atypical CLL morphology, immunophenotype, DNA methylome, and expression profile that differ from conventional CLL. In contrast, downstream BCL3-R neoplasms were atypical splenic or nodal marginal zone lymphomas (MZL) with mutated IGHV, complex karyotypes and mutated genes typical of MZL. Two of the latter four tumors transformed to a large B-cell lymphoma. We designed a novel fluorescence in situ hybridization assay that recognizes the two different breakpoints and validated these findings in 17 independent tumors. Overall, upstream or downstream breakpoints of BCL3-R are mainly associated with two subtypes of lymphoid neoplasms with different (epi)genomic, expression, and clinicopathological features resembling atypical CLL and MZL, respectively