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Continued waves, new variants, and limited vaccine deployment mean that SARS-CoV-2 tests remain vital to constrain the coronavirus disease 2019 (COVID-19) pandemic. Affordable, point-of-care (PoC) tests allow rapid screening in non-medical settings. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) is an appealing approach. A crucial step is to optimize testing in low/medium resource settings. Here, we optimized RT-LAMP for SARS-CoV-2 and human β-actin, and tested clinical samples in multiple countries. “TTTT” linker primers did not improve performance, and while guanidine hydrochloride, betaine and/or Igepal-CA-630 enhanced detection of synthetic RNA, only the latter two improved direct assays on nasopharygeal samples. With extracted clinical RNA, a 20 min RT-LAMP assay was essentially as sensitive as RT-PCR. With raw Canadian nasopharygeal samples, sensitivity was 100% (95% CI: 67.6% - 100%) for those with RT-qPCR Ct values ≤ 25, and 80% (95% CI: 58.4% - 91.9%) for those with 25 < Ct ≤ 27.2. Highly infectious, high titer cases were also detected in Colombian and Ecuadorian labs. We further demonstrate the utility of replacing thermocyclers with a portable PoC device (FluoroPLUM). These combined PoC molecular and hardware tools may help to limit community transmission of SARS-CoV-2.

Celiac disease (CD) is an autoimmune enteropathy induced by the ingestion of gluten from wheat, barley, and rye in genetically susceptible individuals. The global prevalence of CD is 1.4%. However, most of the prevalence studies have been conducted in Caucasian populations; few studies have been performed in Latin America. The aim of this study is to determine the seroprevalence of auto-antibodies used as markers for CD in a Colombian cohort. In this cross-sectional study, the serum samples from Colombian donors of the National Red Cross Blood Bank were collected between June and September 2017 in Bogotá, Colombia. All sera were tested for IgA antitissue transglutaminase (TTG) by enzyme-linked immunosorbent assay. Seropositive sera were tested for IgA antiendomysium (EMA) using indirect immunofluorescence assay. The ancestral genetic composition was determined in donor samples with antibody assay reactivity. Those with two seroreactive assays were typed for HLA class II DQ2 and DQ8. In total, 228 blood donors participated in the study. Among them, 113 were females (49.56%) with an average age of 31.63 years (SD ± 12.99); males had an average of 34.71 years (SD ± 13.01). Only 3 (1.31%) donors reported chronic diarrhea and nonintentional weight loss; 11 (4.82%) had a family history of CD. For the serological assays, 11 donors (4.82%) were seroreactive to IgA anti-TTG: 3 had high reactivity and 8 had low reactivity. Of those seroreactive to IgA anti-TTG, 3 (1.32%) were also seroreactive to anti-EMA, and they were typed as HLA-DQ8 or HLA-DQ2. The baseline ancestral percentage of the seroreactive donors was higher for European and Native American than for African genes. The seroprevalence for anti-TTG and anti-EMA with the presence of HLA-DQ8 and HLA-DQ2 was 1.32%. Additionally, 4.82% donor participants were reactive only for anti-TTG. Compared with other studies, our findings suggest that Colombia has a high prevalence of CD markers.

Abstract Introduction Chagas disease is a parasitic infection whose pathogenesis is related to parasite persistence and a dysfunctional cellular immune response. Variability in cytokine secretion among chronic Trypanosoma cruzi‐infected patients might preclude the identification of the pool of antigen specific T cells. The goal of this study was to determine the fraction of T cells responding to T. cruzi antigen measured by the expression of membrane TNF‐α and CD154. Methods A total of 21 chagasic patients, 11 healthy and 5 non‐chagasic cardiomyopathy controls were analyzed. PBMCs were short‐term cultured in the presence of anti‐CD28, anti‐CD49d, anti‐TNF‐α, and TACE (TNF‐α converting enzyme) inhibitor either under T. cruzi‐lysate or polyclonal stimuli. Cells were stained with anti‐CD3, anti‐CD4, anti‐CD8, and anti‐CD154, and analyzed with flow cytometry. Results CD4+ and CD8+ T cells in chagasic patients displayed higher percentages of membrane‐bound TNF‐α+ and CD154+ compared with controls after T. cruzi‐antigen stimulation. Both markers displayed a positive correlation in the T cell subpopulations analyzed. Symptomatic chagasic patients were differentiated from asymptomatic patients based on the expression of CD154 and membrane TNF‐α in TCD4+ and TCD8+ compartments, respectively. Conclusions These results show that both markers could be useful for assessing the pool of antigen‐specific T cells in chronic chagasic patients.

Monocytes are classified according to their CD14 and CD16 expression into classical (reparative), intermediate (inflammatory), and non-classical. This study assessed the frequency of monocyte and the relationship between monocyte subset percentages and the levels of blood cytokines in Colombian chagasic patients with different clinical forms. This study included chagasic patients in different clinical stages: indeterminate (IND) n = 14, chronic chagasic cardiomyopathy (CCC) n = 14, and heart transplant chagasic (HTCC) n = 9; controls with non-chagasic cardiopathy (NCC) n = 15, and healthy individuals (HI) n = 15. Peripheral blood mononuclear cells (PBMCs) were isolated, labeled for CD14, CD16, and HLA-DR, and analyzed by flow cytometry. Cytokines were measured with a bead-based immunoassay. Percentages of total CD14+ CD16+ and CD14+ HLA-DR+ monocytes were higher in patients with heart involvement (CCC, HTCC, and NCC) than controls. Percentages of intermediate monocytes increased in symptomatic chagasic patients (CCC and HTCC) compared to asymptomatic chagasic patients (IND) and controls (HI). Asymptomatic chagasic patients (IND) had higher percentages of classical monocytes, an increased production of CCL17 chemokine compared to chagasic symptomatic patients (CCC), and their levels of CCL17 was positively correlated with the percentage of classical monocyte subset. In CCC, the percentages of intermediate and classical monocytes were positively correlated with IL-6 levels, which were higher in this group compared to HI, and negatively with IL-12p40 concentration, respectively. Remarkably, there also was an important increased of classical monocytes frequency in three chronic chagasic patients who underwent cardiac transplant, of which one received anti-parasitic treatment. Our findings suggest that cardiac chagasic patients have an increased percentage of inflammatory monocytes and produce more IL-6, a biomarker of heart failure and left ventricular dysfunction, whereas asymptomatic chagasic individuals present a higher percentage of reparative monocytes and CCL17.

Glioblastoma (GBM) is the most prevalent type of primary brain tumor. Treatment options include maximal surgical resection and drug-radiotherapy combination. However, patient prognosis remains very poor, prompting the search for new models for drug discovery and testing, especially those that allow assessment of in vivo responses to treatment. Zebrafish xenograft models have an enormous potential to study tumor behavior, proliferation and cellular interactions. Here, an in vivo imaging and proliferation assessment method of human GBM xenograft in zebrafish larvae is introduced. Zebrafish larvae microinjected with fluorescently labeled human GBM cells were screened daily using a stereomicroscope and imaged by light sheet fluorescence microscopy (LSFM); volumetric modeling and composite reconstructions were done in single individuals. Larvae containing tumors were enzymatically dissociated, and proliferation of cancer cells was measured using dye dilution by flow cytometry. GBM micro-tumors formed mainly in the zebrafish yolk sac and perivitelline space following injection in the yolk sac, with an engraftment rate of 73%. Daily image analysis suggested cellular division, as micro-tumors progressively grew with differentiated fluorescence intensity signals. Using dye dilution assay by flow cytometry, at least three GBM cells' division cycles were identified. The combination of LSFM and flow cytometry allows assessment of proliferation and tumor growth of human GBM inside zebrafish, making it a useful model to identify effective anti-proliferative agents in a preclinical setting.

La cardiopatía chagásica crónica se presenta en un 30% de las personas infectadas con Trypanosoma cruzi. Aunque el parásito puede ser controlado por la respuesta inmune después de la fase aguda, su detección se hace difícil en la fase crónica a pesar de la persistencia de éste en los tejidos de los individuos infectados. Dado que solo un porcentaje de estos individuos crónicamente infectados desarrolla el daño tisular, se sugiere la existencia de factores asociados que predicen la aparición de la sintomatología crónica: a) la genética del hospedero (moléculas del HLA), cuyo papel aún no se ha dilucidado, b) factores dependientes del parásito cómo la variabilidad de los genotipos (TcI a TcVI), su asociación con tropismo y daño tisular; y c) otros factores como la cantidad del inóculo, la reexposición constante a vectores infectados, las diferentes vías de infección y el estado inmunológico del hospedero. Varias teorías han sido implicadas en el compromiso cardiaco, como la persistencia del T. cruzi en los tejidos, la autoinmunidad inducida y el daño tisular producido por la respuesta inmune. En esta revisión se pretende emitir una hipótesis respecto a la disfunción celular inmune producida por la persistencia parasitaria en los tejidos y su papel en la patogénesis de la enfermedad. Se consideran aspectos como el pronóstico de los pacientes con cardiopatía chagásica llevados a trasplante de corazón por falla cardiaca avanzada comparado con otras causas de falla que conducen a trasplante y la posible organización de los infiltrados inflamatorios en el tejido cardiaco, el cual podría funcionar como un tejido linfoide terciario.

Objetivo. Se realizó una búsqueda sistemática de la literatura científica con el objetivo de conocer el número de artículos colombianos publicados con el uso de citometría de flujo, sus aplicaciones y el aporte a la literatura mundial sobre el tema.Método. Se utilizaron las bases de datos electrónicas PubMed, LILACS e ISI Web of Knowledge, y se buscaron los términos “flow cytometry” and “Colombia” o “citometría de flujo” y “Colombia”. Se incluyeron trabajos de investigación experimentales que usaron la citometría de flujo y cuyo primero o último autor pertenecieran a grupos de investigación colombianos.Resultados. Se encontraron 74 artículos que cumplían los criterios de inclusión, publicados desde agosto de 1997 hasta marzo de 2010. La gran mayoría de los trabajos pertenecía al área biomédica y en 54% se aplicó al estudio de enfermedades infecciosas. El número de publicaciones por año aumentó desde 1997 hasta 2004, cuando se presentó un pico de 11 artículos. En total, se encontraron 27 grupos de investigación, pertenecientes a 13 instituciones. El 39,2% (n=29) de los trabajos fueron publicados por grupos de investigación de la Universidad de Antioquia. El tipo de aplicación de la citometría de flujo más usada fue el análisis de inmunofenotipo (58,1%).Conclusiones. Para el periodo comprendido entre agosto de 1997 y marzo de 2010, PubMed reportó 76.312 publicaciones relacionadas con la citometría de flujo. Por consiguiente, el aporte de las publicaciones colombianas a la literatura mundial es tan sólo de 0,096%.

El concepto de “privilegiado” del sistema nervioso central con respecto a la respuesta inmune ha evolucionado en los últimos años debido a la participación de elementos locales en los mecanismos de defensa. Se ha podido demostrar el papel de la neuroglia en la respuesta inmune local en base al estudio de diversas patologías infecciosas y auto-inmunes del sistema nervioso central, tanto en humanos y como en modelos murinos. La neuroglia secreta y es blanco de diferentes citocinas y quimocinas que participan en la respuesta inflamatoria del sistema nervioso ante agresiones de diversos tipos tanto biológicas como físicas. Se ha demostrado la presencia in vivo de moléculas de clase I del complejo mayor de histocompatibilidad en la microglia, los astrocitos y los oligodendrocitos pero sólo durante procesos inflamatorios, la presencia de moléculas de clase II se ha comprobado únicamente en la microglia y su identificación en los astrositos es motivo de controversia. La presencia de algunos marcadores inmunológicos en la neuroglia y los estudios funcionales ha permitido identificar mecanismos como la fagocitosis y la presentación de antígenos a los linfocitos TCD4+ por parte de la microglia, la secreción de citocinas por los astrocitos y la presencia en oligodendrocitos de receptores para estos factores solubles. Todos estos avances han permitido la comprensión de algunos mecanismos fisio-patogénicos y la posible aplicación de novedosas medidas terapéuticas en el sistema nervioso central.

Chagas disease is a parasitic infection caused by the protozoan Trypanosoma cruzi. One of the complications of the disease is the infection of the central nervous system (CNS), as it can result from either the acute phase or by reactivation during the chronic phase, exhibiting high mortality in immunocompromised patients. This systematic review aimed to determine clinical and paraclinical characteristics of patients with Chagas disease in the CNS. Articles were searched from PubMed, Scopus and LILACS until January 2023. From 2325 articles, 59 case reports and 13 case series of patients with Chagas in the CNS were retrieved from which 138 patients were identified. In this population, 77% of the patients were male, with a median age of 35 years old, from which most of them came from Argentina and Brazil. Most of the individuals were immunocompromised from which 89% were HIV-positive, and 54 patients had an average of 48 cells per mm3 CD4+ T cells. Motor deficits and seizures were the most common manifestation of CNS compromise. Furthermore, 90 patients had a documented CNS lesion by imaging from which 89% were supratentorial and 86% were in the anterior/middle cranial fossa. The overall mortality was of 74%. Among patients who were empirically treated with anti-toxoplasma drugs, 70% died. This review shows how Chagas disease in the CNS is a devastating complication requiring prompt diagnosis and treatment to improve patients’ outcomes.

Introduction: The objective of this study was to examine the effects of in ovo nicotinamide riboside (NR) feeding on high-yield broiler growth and meat quality.Methods: Fertilized Cobb 700 by-product eggs (N = 3,240) were randomly assigned to one of four in ovo treatments and injected with 0 (0NR), 250 (250NR), 500 (500NR), or 1,000 (1,000NR) mM NR at incubation-day 10. Chicks were hatched, vent sexed, and randomly placed 18 per pen in one of 32 floor pens. On day 48, birds were processed and deboned.Results: There were dose effects for all part weights (p < 0.05). Pectoralis major weight of 250, 500, and 1,000NR carcasses were heavier than 0NR (p < 0.03) but did not differ from remaining NR doses (p > 0.26). Pectoralis minor weight of 250NR carcasses was greater (p < 0.01) than 0NR and did not differ from other NR tenders (p > 0.21). Pectoralis minor weight of 500 and 1,000NR carcasses was greater than 0NR (p < 0.09), but did not differ (P = 0.82) from each other. There were no dose effects for all Pectoralis major and minor myopathy scores and incidence except incidence of tenders scoring “0” and “1” for woody-like tender. Percentage of NR1,000 tenders scoring 0 and 1 for woody-like tender were less than and greater than all other treatments, respectively (p < 0.05). There were no differences among remaining NR doses and NR0 tenders (p > 0.10). There were dose effects for muscle fiber number (P = 0.03). There tended to be more muscle fibers within 250 and 1,000NR muscles compared to 0NR (p < 0.09). Pectoralis major muscle from 500NR did not differ in muscle fiber number compared to 250 and 1,000NR (p > 0.18), but had more (p < 0.01) fibers than 0NR muscle. There tended to be more fibers in 250 and 1,000NR muscles compared to 0NR muscle (p < 0.09).Discussion: Nicotinamide riboside in ovo feeding caused birds to produce heavier parts; however, myopathy scores and incidence were minimally affected which may have been due greater muscle fiber number.

The objective of this study was to determine the daily dietary nicotinamide riboside (NR) dose required to maximize the delay of subjective muscle fatigue onset. Barrows (N = 100) were assigned to one of five treatments: a conventional swine finishing diet containing 0 (CON), 15 (15NR), 30 (30NR), 45 (45NR) mg·kg body weight−1·d−1 NR, or CON supplemented with 45 mg·kg body weight−1·d−1 NR by drench or cookie dough (DRE). All treatments were administered for the final 11 days of feeding. On supplementation d 10, barrows individually experienced a performance test at 1.09 m/s until they were subjectively exhausted. Wireless electromyography (EMG) sensors were affixed to the biceps femoris (BF), tensor fascia latae (TFL), and semitendinosus (ST) to measure real-time muscle activity. There were no treatment effects for barrow speed (p = 0.57), a tendency for a treatment effect (p = 0.07) for distance, and a treatment effect (p = 0.04) on time to exhaustion. Barrows of the 15NR and DRE treatments had greater (p = 0.05) distances to exhaustion than CON barrows but did not differ from other NR barrows (p > 0.11). Barrows in the 45NR treatment did not differ (p = 0.11) in distance from 30NR barrows but tended to have a greater (p = 0.07) distance compared to CON barrows. All other treatment comparisons did not differ (p > 0.27). Barrows in the DRE treatment moved for longer (p < 0.01) than CON barrows, but all other treatments did not differ from each other (p > 0.15). There was no treatment × period interaction for all muscles’ root mean square (RMS) values (p > 0.16), but there were Period effects for all muscles (p < 0.01) and a Treatment effect (p = 0.04) in the TFL. For all muscles, period 4 had greater RMS values than all other periods (p < 0.01), who did not differ from each other (p > 0.29). In the TFL, CON barrows had greater RMS values during the performance test compared to all NR treatments (p < 0.02), who did not differ from each other (p > 0.18). Overall, NR demonstrates potential in being a useful tool in fatigue prevention, but efficient administration of the compound needs further investigation.

Trypanosoma cruzi, the causal agent of Chagas disease, has coexisted with humans for thousands of years. Therefore, the parasite has developed several mechanisms of antigenic variability that has allowed it to live inside the cells and evade the host immune response. Since T. cruzi displays an intracellular cycle-stage, our research team focused on providing insights into the CD8+ T cells immune response in chronic Chagas cardiomyopathy. We began our work in the 2000s studying parasite antigens that induce natural immune responses such as the KMP11 protein and TcTLE, its N-terminal derived peptide. Different approaches allowed us to reveal TcTLE peptide as a promiscuous CD8+ T cell epitope, able of inducing multifunctional cellular immune responses and eliciting a humoral response capable of decreasing parasite movement and infective capacity. Next, we demonstrated that as the disease progresses, total CD8+ T cells display a dysfunctional state characterized by a prolonged hyper-activation state along with an increase of inhibitory receptors (2B4, CD160, PD-1, TIM-3, CTLA-4) expression, an increase of specific terminal effector T cells (TTE), a decrease of proliferative capacity, a decrease of stem cell memory (TSCM) frequency, and a decrease of CD28 and CD3ζ expression. Thus, parasite-specific CD8+ T cells undergo clonal exhaustion, distinguished by an increase in late-differentiated cells, a mono-functional response, and enhanced expression of inhibitory receptors. Finally, it was found that anti-parasitic treatment induces an improved CD8+ T cell response in asymptomatic individuals, and a mouse animal model led us to establish a correlation between the quality of the CD8+ T cell responses and the outcome of chronic infection. In the future, using OMICs strategies, the identification of the specific cellular signals involved in disease progression will provide an invaluable resource for discovering new biomarkers of progression or new vaccine and immunotherapy strategies. Also, the inclusion of the TcTLE peptide in the rational design of epitope-based vaccines, the development of immunotherapy strategies using TSCM or the blocking of inhibitory receptors, and the use of the CD8+ T cell response quality to follow treatments, immunotherapies or vaccines, all are alternatives than could be explored in the fight against Chagas disease.

The objective of this study was to collect and quantify three-axis acceleration data from six locations within commercial pig transport trailers during summer. Two trucks with straight-deck trailers transporting two loads per day were observed for 5 consecutive days (N = 20). Accelerometers were placed under the floor of each trailer’s top and bottom decks (DECs) in the center of three sections (SECs): fore, middle, and aft. Data from each trailer section were processed to calculate z- and x,y-axis root mean square (RMS) values and vibration dose values (VDVs) during loading, transport, and unloading. There were no DEC × SEC interactions or SEC main effects for z-axis RMS or VDV during any transportation stage (P > 0.06). The bottom deck had a greater x,y-axis RMS than the top deck during all transportation stages (P < 0.01). The bottom deck had a greater x,y-axis VDV than the top deck during loading and transport (P < 0.03), but there was no difference (P = 0.52) during unloading. The bottom deck had a greater z-axis RMS and VDV than the top deck during loading and transport (P < 0.01), but there were no differences during unloading (P > 0.07). There were no SEC effects for x,y- and z-axis RMSs and VDVs during all transportation stages (P > 0.06). Acceleration values were compared with exposure action values (EAV; injury possible) and exposure limit values (ELV; injury likely) vibrations thresholds. Over the 5 observation days during all transport stages, a greater percentage of compartments violated both RMS and VDV thresholds in the x,y orientation (average 90%) than in the z orientation (average 76%). Overall, these data indicate that bottom decks experience greater three-axis vibrations than top decks in straight-deck trailers and that pigs on bottom decks may experience greater discomfort during transportation that could contribute to fatigue or the non-ambulatory condition.

Background Characterized by an inflammatory pathogenesis, acne is the most common skin disorder worldwide. Altered sebum production, abnormal proliferation of keratinocytes, and microbiota dysbiosis represented by disbalance in Cutibacterium acnes population structure, have a synergic effect on inflammation of acne-compromised skin. Although the role of C. acnes as a single factor in acne development is still under debate, it is known that skin and skin-resident immune cells recognize this bacterium and produce inflammatory markers as a result. Control of the inflammatory response is frequently the target for acne treatment, using diverse chemical or physical agents including antibiotics. However, some of these treatments have side effects that compromise patient adherence and drug safety and in the case of antibiotics, it has been reported C. acnes resistance to these molecules. Phage therapy is an alternative to treat antibiotic-resistant bacterial strains and have been recently proposed as an immunomodulatory therapy. Here, we explore this perspective about phage therapy for acne, considering the potential immunomodulatory role of phages. Methodology Literature review was performed using four different databases (Europe PubMed Central-ePMC, Google Scholar, PubMed, and ScienceDirect). Articles were ordered and selected according to their year of publication, number of citations, and quartile of the publishing journal. Results The use of lytic bacteriophages to control bacterial infections has proven its promising results, and anti-inflammatory effects have been found for some bacteriophages and phage therapy. These effects can be related to bacterial elimination or direct interaction with immune cells that result in the regulation of pro-inflammatory cytokines. Studies on C. acnes bacteriophages have investigated their lytic activity, genomic structure, and stability on different matrices. However, studies exploring the potential of immunomodulation of these bacteriophages are still scarce. Conclusions C. acnes bacteriophages, as well as other phages, may have direct immunomodulatory effects that are yet to be fully elucidated. To our knowledge, to the date that this review was written, there are only two studies that investigate anti-inflammatory properties for C. acnes bacteriophages. In those studies, it has been evidenced reduction of pro-inflammatory response to C. acnes inoculation in mice after bacteriophage application. Nevertheless, these studies were conducted in mice, and the interaction with the immune response was not described. Phage therapy to treat acne can be a suitable therapeutic alternative to C. acnes control, which in turn can aid to restore the skin’s balance of microbiota. By controlling C. acnes colonization, C. acnes bacteriophages can reduce inflammatory reactions triggered by this bacterium.

Objective: CD4+CD8+ double-positive T-cells (DPTs) have been classified as a separate T-cell subpopulation, with two main phenotypes: CD4high CD8low and CD4low CD8high. In recent years, the relevance of DPTs in the pathogenesis of infections, tumors, and autoimmune diseases has been recognized. Reference values among healthy individuals remain unknown. Therefore, the aim of this study is to provide a reference value for DPTs in peripheral blood from healthy donors in a blood bank in Bogotá, Colombia, and to determine the activation status using a surface marker. Materials and Methods: One hundred healthy donors were enrolled in the study. Peripheral blood cells were stained for CD3, CD4, CD8, and CD154 (CD40L), and cellular viability was assessed with 7-aminoactinomycin D and analyzed by flow cytometry. Results: The median value for DPTs was 2.6% (interquartile range=1.70%-3.67%). Women had higher percentages of DPTs than men (3.3% vs. 2.1%). The subpopulation of CD4low CD8high showed higher expression of CD154 than the other T-cell subpopulations. Conclusion: DPT reference values were obtained from blood bank donors. A sex difference was found, and the CD4low CD8high subpopulation had the highest activation marker expression.

¿Por qué dependemos de la biodiversidad? Autor: Juan Armando Sánchez Muñoz ISBN: 978-958-504-022-9 Editorial: Intermedio Editores Ediciones Uniandes Fecha de publicación: Noviembre del 2021 198 pp.

La mayoría de las evaluaciones serológicas comunitarias específicas para los anticuerpos de SARSCoV-2 se han realizado en instituciones y en trabajadores de la salud. En este estudio, se evaluaron anticuerpos IgG específicos para el virus en personas que trabajaban en una universidad de Bogotá, Colombia. El objetivo fue determinar la exposición previa a SARS-CoV-2 en los asistentes al campus durante el cierre de la ciudad. Se evaluaron 237 personas, incluidas 93 mujeres y 144 hombres, mediante la detección quimioluminiscente de anticuerpos IgG anti-proteína N viral entre noviembre y diciembre de 2020. Se encontraron 32 individuos positivos, es decir, una seroprevalencia del 13,5 % (10 mujeres y 22 hombres), en su mayoría asintomáticos (68,75 %) y se determinaron tres grupos de individuos seropositivos. Solo 13 de los individuos serorreactivos tenían una detección positiva previa del ARN del SARS-CoV-2 mediante RT-qPCR realizada, en promedio, 91 días antes de la prueba serológica. Los individuos seropositivos no provenían de las localidades con los porcentajes más altos de casos de SARS-CoV-2 en la ciudad. La encuesta se realizó después del primer pico de transmisión del SARS-CoV-2 en la ciudad y antes de la preparación para la reapertura del campus a estudiantes en el 2021, y demostró una baja seroprevalencia con un alto procentaje de asintomáticos. Estos resultados ayudarán a evaluar algunas de las estrategias establecidas para controlar la propagación del virus en instituciones educativas u otras comunidades similares.

The objective of this study was to determine the effects of in ovo injection of nicotinamide riboside (NR) on broiler embryonic myogenesis. Fertilized Cobb 500 broiler eggs (N = 240) were sorted by weight and within each strata, randomly assigned to 1 of 4 NR dose treatments (0 mmol, 250 mmol, 500 mmol, or 1 mol; final concentration in yolk of 0, 2.5, 5.0, or 10.0 mmol) of NR. At day 10 of incubation, 100 μL of the assigned NR dose was injected into the yolk sac of the developing embryo, and chicks were euthanized within 24 h of hatching. Pectoralis major muscle (PMM) and individual fiber morphometrics were collected. Chicks injected with NR had greater PMM weight and length (P 0.14). Chicks from eggs injected with NR had greater PMM weight and width than control chicks (P 0.06). There was no treatment effect (P = 0.20) for PMM fiber cross-sectional area; however, there was a treatment effect (P

The objective of this study was to evaluate the repeatability of the Pressed Juice Percentage (PJP) as well as to determine the accuracy of previously determined PJP thresholds at sorting beef strip loin steaks into categories based on the probability of being rated juicy by sensory panelists. Beef strip loin steaks representing 3 USDA quality grades (Prime, Low Choice, and Low Select) and Low Select strip loin steaks enhanced to 108% of the raw weight with a water, salt, and alkaline phosphate solution were cooked to three degrees of doneness [DOD; Rare (60°C), Medium (71°C), Very Well-Done (82°C)] to maximize variation in juiciness. Paired steaks were evaluated for PJP, Slice Shear Force (SSF), and Warner-Bratzler shear force (WBSF). Additionally, steaks representing Prime, Low Choice, and Low Select that were non-enhanced or enhanced with the same enhancement solution cooked to three DOD [Rare (60°C), Medium (71°C), Very Well-Done (82°C)] from a previously conducted sensory study were used to evaluate the ability of PJP to sort steaks into juiciness categories. Results indicated PJP had a high repeatability coefficient of 0.70, indicating only a small portion (30%) of the variation observed was due to sample measurement differences between paired samples. The repeatability of SSF and WBSF were 0.68 and 0.85, respectively. The evaluated PJP threshold values accurately segregated strip loin steaks by the probability of a sample being rated as juicy (mean juiciness rating of > 50) by consumers. The actual percentage of juicy samples was determined to be 41.67, 72.31, 89.33, and 98.08% for the predicted < 50%, 50 to 75%, 75 to 90%, and > 90% categories, respectively. Results of this study indicate the PJP juiciness measure is both repeatable and accurate in sorting beef strip loin steaks based on the likelihood of a steak being classified as juicy by consumers.

The objective of this study was to evaluate the impact of marbling level in combination with enhancement on beef palatability when strip loin steaks were cooked to 3 degrees of doneness. Consumer and trained sensory panelists evaluated strip steak palatability traits of 3 USDA quality grades: Prime, Low Choice, and Low Select. Additional strip loins from each grade were enhanced to 108% with a water, salt, and alkaline phosphate solution. Steaks from each treatment were cooked to 3 degrees of doneness (DOD; Rare: 60°C, Medium: 71°C, or Very Well-Done: 82°C). Consumer panelists rated all enhanced treatments similar ( > 0.05) for each palatability trait. Enhanced steaks had greater ( < 0.05) juiciness, tenderness, flavor liking, and overall liking ratings than all non-enhanced treatments, regardless of grade. Consumer juiciness, tenderness, and overall liking scores increased ( < 0.05) as DOD decreased. Trained sensory panelists rated all enhanced treatments similar ( > 0.05) for initial and sustained juiciness, myofibrillar tenderness, and overall tenderness and greater ( < 0.05) than all non-enhanced treatments, other than non-enhanced Prime, for the same traits. Trained sensory panel ratings increased ( < 0.05) as DOD decreased for sustained juiciness, and both myofibrillar and overall tenderness. Moreover, Warner-Bratzler shear force values were similar ( > 0.05) among all enhanced treatments, and lower ( < 0.05) than non-enhanced Low Choice and Low Select treatments. Results from this study indicate marbling level has no impact on the palatability traits of enhanced strip loin steaks. Therefore, enhancement of higher valued, high marbled cuts does not provide additional palatability benefits over low marbled cuts, as enhancement does not provide an additive effect with marbling on beef eating quality.

The objective of this study was to assess consumer and chef perceptions and knowledge of beef degrees of doneness (DOD) as well as to measure the changes in cooked color over time related to DOD. Steaks from strip loins (M. longissimus lumborum) from each of 5 quality treatments were used for this study. Steaks were cooked to an endpoint temperature of either very-rare (54°C), rare (60°C), medium-rare (63°C), medium (71°C), well-done (77°C), or very well-done (82°C). L*, a*, and b* were evaluated at 0, 1, 2, 3, 6, 9, and 12 min post-cutting and digital pictures were taken immediately on an internal surface of the steak. Digital surveys for the evaluation of the images of the cooked steaks were created for consumers and chefs. There were time × DOD interactions (P < 0.05) for L*, a*, and b* values. For very-rare, rare, medium-rare, and medium, a* values increased (P < 0.05) over time. For L*, well-done and very well-done steaks became darker (P < 0.05) and very-rare, rare, and medium-rare steaks became lighter (P < 0.05) over time. Surveys indicated consumers determine DOD when cooking beef in their home primarily by using color, whereas chefs primarily use touch to determine DOD. There were no quality grade effects (P > 0.05) for DOD responses for steak pictures evaluated by consumers or chefs. Consumers identified the DOD of cooked steaks as the DOD that corresponds to published end-point temperatures 27 to 35% of the time. Chefs typically identified the DOD as 1 DOD higher than which the steaks were cooked for steaks cooked to medium or less and 1 DOD lower for steaks cooked to well-done and higher. This indicates differences exist in the perceptions of DOD between culinary professionals and consumers, and may contribute to decreased consumer satisfaction when ordering steaks in a restaurant.

The objective of this study was to evaluate the effect of increased pork hot carcass weights on consumer visual acceptability and purchase intent ratings of top loin chops cut to various thicknesses in a price labeled versus unlabeled retail display scenario. Pork loins (N = 200) were collected from 4 different hot carcass weight groups: light weight (LT; less than 111.8 kg), medium-light weight (MLT; 111.8 to 119. kg), medium-heavy weight (MHVY; 119.1 to 124.4 kg), and a heavy weight group (HVY; 124.4 kg and greater). Loins were fabricated into 4 pairs of chops of specified thicknesses (1.27, 1.91, 2.54, and 3.18 cm). One chop from each pair was assigned to be packaged with or without a label. Consumers assessed chops for appearance, desirability, and purchase intent. For both appearance and purchase intent ratings, chops from HVY carcasses were given more desirable (P < 0.05) ratings compared to LT chops. Consumers gave greater (P < 0.05) appearance ratings to thicker cut chops. There was a hot carcass weight × chop thickness interaction (P < 0.05) for the percentage of consumers that indicated the chop was desirable overall. Regardless of hot carcass weight group, chops with a thickness of 1.27 cm had the lowest (P < 0.05) percentage of consumers indicate they were desirable overall. A greater (P < 0.05) percentage of consumers indicated “yes” they would purchase chops cut to a thickness of 2.54 cm compared to all other thicknesses. Additionally, there was a greater (P < 0.05) percentage of consumers who indicated they would purchase unlabeled chops compared to labeled chops. These results, within the population sampled, indicate that carcass weight and chop thickness can affect consumer preference and thus should be considered by retailers when marketing fresh pork loin chops.

Aerobically packaged frozen bacon has significant challenges to flavor and odor properties as storage length advances. Naturally smoked sugar (NSS), a food ingredient made from applying hardwood smoke to liquid sugar, may possess antioxidant functionality that is beneficial in controlling lipid oxidation in bacon. Therefore, the objective of this study was to determine if NSS could be added directly to a bacon formulation to limit the rate of lipid oxidation in frozen, aerobically packaged bacon. Three replications of this experiment were conducted using 5 pork bellies per replication. Individual bellies were cut in half yielding an anterior and posterior section and then randomly assigned to a treatment combination with either the anterior or posterior section receiving the NSS treatment. Treatment brines consisted of a control (CON) brine or a brine with added NSS injected to retain 12% added solution. After injection, smoking, cooking, and slicing, bacon slices were frozen (–17.8 ± 2°C) and stored aerobically for 0, 40, 80, and 120 d for sensory and gas chromatography mass spectrometry (GCMS) analyses or 0, 20, 40, 60, 80, 100, and 120 d for thiobarbituric acid reactive substances (TBARS) analysis. There were significant (P < 0.01) Treatment × Day interactions for oxidized flavor intensity, TBARS, and hexanal concentration. Panelist oxidized flavor intensity scores, TBARS values, and hexanal content increased from d 0 to 120 (P < 0.01) for CON, whereas these measures in bacon manufactured with added NSS did not change (P > 0.16). Sensory ratings for saltiness, smoke intensity, and bacon flavor intensity were higher (P < 0.01) for the NSS treatment compared to CON. The ability of NSS to function as an effective antioxidant in frozen bacon was confirmed by the inhibition of lipid oxidation products and improved sensory panel scores over time.

The objective of this study was to evaluate the effects of increased pork hot carcass weight on loin quality and palatability of top loin chops. Pork loins (N = 200) were collected from 4 different hot carcass weight groups: A light weight (LT; less than 111.8 kg), medium-light weight (MLT; 111.8 to 119.1 kg), medium-heavy weight (MHVY; 119.1 to 124.4), and a heavyweight group (HVY; 124.4 and greater). Following fabrication, chops were assigned to fat and moisture analysis, Warner-Bratzler shear force (WBSF), consumer sensory panels, or trained sensory panels. Chops from the HVY group were rated as more (P < 0.05) tender compared to chops from the LT carcasses. Additionally, chops from the HVY weight group had greater (P < 0.05) consumer overall like ratings compared to chops from both the LT and MLT groups. Carcass weight did not affect (P > 0.05) consumer flavor liking ratings. Hot carcass weight treatment did not contribute (P > 0.05) to the percentage of chops rated acceptable for flavor and overall liking. The greatest (P < 0.05) percentage of samples were rated acceptable for juiciness for chops from the HVY weight group, and the lowest (P < 0.05) percentage of acceptable ratings for tenderness were for chops from the LT weight group. Both initial and sustained juiciness from MHVY carcasses were rated as more (P < 0.05) juicy compared to chops from both MLT and LT carcasses by trained sensory panelists. Additionally, chops from the LT carcasses had the lowest (P < 0.05) myofibrillar tenderness ratings. Chops from MHVY and HVY carcasses were similar (P > 0.05), with greater (P < 0.05) overall tenderness ratings compared to chops from LT carcasses. These results indicate chops from heavier weight carcasses may have improved tenderness and juiciness compared to chops from lighter carcasses.

The objective of this study was to evaluate the impact of USDA quality grade on beef eating quality of top sirloin steaks when cooked to multiple degrees of doneness (DOD). Beef top sirloin butts (N = 60; 15/quality grade) were collected to equally represent 4 quality grades [Prime, Top Choice (modest00 to moderate100), Low Choice, and Select]. Steaks were assigned to 1 of 3 DOD: rare (60°C), medium (71°C), and well-done (77°C). Steaks were allocated to either consumer sensory analysis, trained sensory analysis, fat and moisture analysis, or Warner-Bratzler shear force (WBSF). There were no interactions (P > 0.05) for all consumer ratings of palatability traits, indicating increases in DOD had the same impact across all quality grades. Prime steaks had greater (P < 0.05) juiciness ratings than all other quality grades, except for Top Choice. As DOD increased, consumer ratings and the percentage of steaks rated acceptable for each palatability trait decreased (P < 0.05; rare > medium > well-done). There was a quality grade × DOD interaction (P < 0.05) for trained sensory panel juiciness scores. When cooked to medium, Prime and Top Choice steaks were rated higher (P < 0.05) for juiciness than Low Choice and Select steaks, while there were no differences at all other DOD. Similar to consumer ratings, trained panel ratings of tenderness decreased (P < 0.05) as DOD increased (rare > medium > well-done). Lastly, there was no quality grade by DOD interaction (P > 0.05) for Warner-Bratzler shear force. These results indicate that regardless of the DOD top sirloin steaks are cooked to, quality grade had minimal impact on palatability. Therefore, it is unnecessary for consumers, retailers, and foodservice to pay premium prices for higher quality top sirloin steaks, as the same eating experience will be provided.

The objective of this study was to determine the impact on beef palatability perceptions when consumers with varying degree of doneness (DOD) preferences are served steaks cooked to multiple DOD. Paired Low Choice strip loin steaks were randomly assigned to a DOD of either rare (60°C), medium-rare (63°C), medium (71°C), medium-well (74°C), or well-done (77°C). Consumer panelists were prescreened for DOD preference (rare, medium, or well-done) prior to sensory panels and were assigned to panels based on their DOD preference. In the first round of testing, consumers were served 1 sample from each of the 5 DOD under low-intensity red incandescent light to mask any DOD differences among samples. In round 2 of testing, consumers were fed the paired samples cooked to the same DOD under white incandescent lights. There were no (P > 0.05) consumer DOD preference × steak DOD interactions or consumer DOD preference effects for tenderness, juiciness, and flavor ratings when steaks were evaluated under both lighting types. Within the white-lighting testing, there was a consumer DOD preference × steak DOD interaction (P < 0.05) for overall acceptability. Consumers who preferred steaks cooked to well-done reported no differences (P > 0.05) in overall palatability among DOD under white-lighting, whereas consumers who preferred steaks cooked to rare and medium rated steaks lower (P < 0.05) for overall palatability as DOD increased. Regardless of DOD preference, consumer sensory ratings decreased (P < 0.05) when steaks were cooked above the consumer’s preferred DOD; whereas sensory ratings improved (P < 0.05) when steaks were served below the consumers’ preferences. These results indicate that overcooking steaks has the greatest negative impact on beef palatability perception and thus, foodservice should err on the side of undercooking steaks to preserve, and potentially improve, eating satisfaction.

Acute leukemia is a heterogeneous set of diseases affecting children and adults. Current prognostic factors are not accurate predictors of the clinical outcome of adult patients and the stratification of risk groups remains insufficient. For that reason, this study proposes a multifactorial analysis which integrates clinical parameters, ex vivo tumor characterization and behavioral in vivo analysis in zebrafish. This model represents a new approach to understand leukemic primary cells behavior and features associated with aggressiveness and metastatic potential. Xenotransplantation of primary samples from patients newly diagnosed with acute leukemia in zebrafish embryos at 48 hpf was used to asses survival rate, dissemination pattern, and metastatic potential. Seven samples from young adults classified in adverse, favorable or intermediate risk group were characterized. Tumor heterogeneity defined by Leukemic stem cell (LSC) proportion, was performed by metabolic and cell membrane biomarkers characterization. Thus, our work combines all these parameters with a robust quantification strategy that provides important information about leukemia biology, their relationship with specific niches and the existent inter and intra-tumor heterogeneity in acute leukemia. In regard to prognostic factors, leukemic stem cell proportion and Patient-derived xenografts (PDX) migration into zebrafish were the variables with highest weights for the prediction analysis. Higher ALDH activity, less differentiated cells and a broader and random migration pattern are related with worse clinical outcome after induction chemotherapy. This model also recapitulates multiple aspects of human acute leukemia and therefore is a promising tool to be employed not only for preclinical studies but also supposes a new tool with a higher resolution compared to traditional methods for an accurate stratification of patients into worse or favorable clinical outcome.

BACKGROUND: Chronic persistent infections have been associated with T lymphocytes functional impairment. The aim of this study was to compare the activation status, the proliferative potential and the expression of CD28 and CD3ζ chain on T lymphocytes between chronic chagasic patients and uninfected controls. METHODOLOGY/PRINCIPAL FINDINGS: Forty-two chronic chagasic patients, 28 healthy individuals and 32 non-chagasic cardiomyopathy donors were included. Peripheral blood was marked for CD3, CD4, CD8, HLA-DR, CD28, CD38 and intracellular CD3ζ. Peripheral blood mononuclear cells were stained with carboxyfluorescein diacetate succinimidylester and incubated with T. cruzi lysate or phytohemagglutinin for five days. Cells from 3 healthy controls were incubated with T. cruzi trypomastigotes separated with transwells; and the expression of CD3ζ chain and proliferation index was determined. Heart-infiltrating cells from two chronic chagasic patients were tested for the aforementioned cellular markers. Chagasic patients displayed higher frequencies of CD4+/HLA-DR+/CD38+ (8.1% ± 6.1) and CD8+/HLA-DR+/CD38+ (19.8 ± 8.9) T cells in comparison with healthy (1.6 ± 1.0; 10.6 ± 8.0) and non-chagasic cardiomyopathy donors (2.9 ± 2.9; 5.8 ± 6.8). Furthermore, the percentage of CD4+ activated T cells was higher in chagasic patients with cardiac involvement. CD8+ T cells proliferation index in chagasic donors (1.7 ± 0.3) was lower when compared with healthy (2.3 ± 0.3) and non-chagasic cardiomyopathy individuals (3.1 ± 1.1). The frequencies of CD4+/CD28+ and CD8+/CD28+ T cells, as well as the CD3ζ(bright)/CD3ζ(dim)% ratios in CD4+ and CD8+ were lower in chagasic patients when compared with both control groups. The CD3ζ(bright)/CD3ζ(dim)% ratio and proliferative indexes for CD4+ and CD8+ T lymphocytes decreased gradually in those cells cultivated with parasites and displayed lower values than those incubated with medium alone. Finally, heart-infiltrating T cells from two T. cruzi infected patients also expressed activation markers and down-regulate CD28 and CD3ζ. CONCLUSIONS: CD8+ T lymphocytes from chagasic donors displayed reduced proliferative capacity, which might be associated with CD3ζ down-regulation and diminished CD28 expression on CD4 T cells.

BACKGROUND: CD4+/CD8+ double positive (DP) T cells have been described in healthy individuals as well as in patients with autoimmune and chronic infectious diseases. In chronic viral infections, this cell subset has effector memory phenotype and displays antigen specificity. No previous studies of double positive T cells in parasite infections have been carried out. METHODOLOGY/PRINCIPAL FINDINGS: Seventeen chronic chagasic patients (7 asymptomatic and 10 symptomatic) and 24 non-infected donors, including 12 healthy and 12 with non-chagasic cardiomyopathy donors were analyzed. Peripheral blood was stained for CD3, CD4, CD8, HLA-DR and CD38, and lymphocytes for intracellular perforin. Antigen specificity was assessed using HLA*A2 tetramers loaded with T. cruzi K1 or influenza virus epitopes. Surface expression of CD107 and intracellular IFN-γ production were determined in K1-specific DP T cells from 11 chagasic donors. Heart tissue from a chronic chagasic patient was stained for both CD8 and CD4 by immunochemistry. Chagasic patients showed higher frequencies of DP T cells (2.1% ± 0.9) compared with healthy (1.1% ± 0.5) and non-chagasic cardiomyopathy (1.2% ± 0.4) donors. DP T cells from Chagasic patients also expressed more HLA-DR, CD38 and perforin and had higher frequencies of T. cruzi K1-specific cells. IFN-γ production in K1-specific cells was higher in asymptomatic patients after polyclonal stimulation, while these cells tended to degranulate more in symptomatic donors. Immunochemistry revealed that double positive T cells infiltrate the cardiac tissue of a chagasic donor. CONCLUSIONS: Chagasic patients have higher percentages of circulating double positive T cells expressing activation markers, potential effector molecules and greater class I antigenic specificity against T. cruzi. Although K1 tetramer positive DP T cell produced little IFN-γ, they displayed degranulation activity that was increased in symptomatic patients. Moreover, K1-specific DP T cells can migrate to the heart tissue.