Functional electrical stimulation to enhance reactive balance among people with hemiparetic stroke.

BACKGROUND: Individuals with stroke demonstrate a twofold higher fall incidence compared to healthy counterparts, potentially associated with deficits in reactive balance control, which is crucial for regaining balance from unpredictable perturbations to the body. Moreover, people with higher stroke-related motor impairment exhibit greater falls and cannot recover balance during higher perturbation intensities. Thus, they might need supplemental agents for fall prevention or even to be included in a perturbation-based protocol. Functional electrical stimulation is a widely used clinical modality for improving gait performance; however, it remains unknown whether it can enhance or interfere with reactive balance control. METHODS: We recruited twelve ambulatory participants with hemiparetic stroke (61.48 ± 6.77 years) and moderate-to-high motor impairment (Chedoke-McMaster Stroke Leg Assessment ≤ 4/7). Each participant experienced 4 unpredicted paretic gait-slips, with and without functional electrical stimulation (provided 50-500 ms after perturbation) in random order. The paretic quadriceps muscle group was chosen to receive electrical stimulation, considering the role of support limb knee extensors for preventing limb-collapse. Outcomes including primary (laboratory falls), secondary (reactive stability, vertical limb support) and tertiary (compensatory step length, step initiation, execution time) measures were compared between the two conditions. RESULTS: Participants demonstrated fewer falls, higher reactive stability, and higher vertical limb support (p < 0.05) following gait-slips with functional electrical stimulation compared to those without. This was accompanied by reduced step initiation time and a longer compensatory step (p < 0.05). CONCLUSION: The application of functional electrical stimulation to paretic quadriceps following gait-slips reduced laboratory fall incidence with enhanced reactive balance outcomes among people with higher stroke-related motor impairment. Our results lay the preliminary groundwork for understanding the instantaneous neuromodulatory effect of functional electrical stimulation in preventing gait-slip falls, future studies could test its therapeutic effect on reactive balance. Clinical registry number: NCT04957355.

Occurrence of Neofusicoccum parvum Causing Canker and Branch Dieback of European Hazelnut in Maule Region, Chile.

The European hazelnut (Corylus avellana) is an important fruit crop cultivated in Chile, with over 17,000 ha planted (46%) in the Maule region, central Chile. During a routine orchard survey in seasons 2020-2021 and 2021-2022, in the Maule region, canker and dieback symptoms were observed in two commercial orchards of European hazelnut cv. Tonda Di Giffoni in San Rafael (8-year-olds) and Linares (15-year-olds), with an incidence between 10% and 36%, respectively, based on external symptoms. Twenty symptomatic branches exhibiting cankers, reduced vigor, wilting, twig death, and dieback, were collected. A cross-section of diseased branches revealed mostly brown V or U-shaped cankers of hard consistency. Branches were cut, and pieces of cankers were surface sterilized in 96% ethanol for 3 s and briefly flamed. Small pieces of wood (5 mm2) from the edge of cankered tissues were placed on Potato Dextrose Agar (2% PDA) amended with 0.1% Igepal CO-630 and incubated at 25°C for five days in the dark (Díaz and Latorre 2014). Pure cultures were obtained by transferring a hyphal tip from growing colonies to fresh PDA media. Eight pure cultures (NP-Haz01 to NP-Haz08) developed dark to olive-brown fast-growing colonies with scarce aerial mycelium after seven days at 25°C on PDA under near-UV light. These isolates showed a dark-olive color on the reverse side of Petri dishes and developed abundant, aggregated, and dark-brown globose pycnidia after 21 days at 25°C. Conidia were hyaline, aseptate, ellipsoidal, densely granulate, externally smooth, and thin-walled dark, that measured (9.5-) 15.5 ±1.2 (-17.3) x (5.1-) 7.2 ± 0.6 (-9.1) µm (n = 30), with a length/width ratio of 2.15. These isolates were tentatively identified morphologically as Neofusicoccum sp. Molecular identification was performed using ITS1/ITS4, Bt2a/Bt2b and EF1-728F/EF1-986R primers of the internal transcribed spacer (ITS1-5.8S-ITS2) region, a portion of the beta-tubulin (BT) and part of the translation elongation factor (EF1-) genes, respectively (Dissanayake et al. 2015). A MegaBlast search in GenBank showed a 99% similarity to isolate CMW9081, the ex-type of Neofusicocum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips. The sequences were added to GenBank (OR393855 to OR393857 for ITS; OR400688 to OR400690 for BT; OR400691 to OR400693 for EF1-). Pathogenicity of three isolates (NP-Haz02, NP-Haz04, NP-Haz09) was studied on freshly made pruning wounds on attached branches of 3-year-old and one-year-old of European hazelnut cv. Tonda Di Giffoni in the San Rafael field. Fifteen pruning wounds were inoculated with 40 µL conidial suspension (105 conidia/mL) of each isolate of N. parvum. Sterile distilled water was used as a control treatment (n=15 branches) for branches of 3-year-olds and one-year-olds. Both pathogenicity tests were repeated once. Attached branches of 3-year-olds (6 months of incubation) and one-year-olds (4 months of incubation), developed necrotic streaks and cankers with a mean length of 33 to 82 mm and 25 to 51 mm, respectively. No necrotic streaks were observed in the branches treated with water. Neofusicoccum parvum was reisolated only from symptomatic tissues of inoculated branches, and morphological and molecularly (EF1-) identified, thus fulfilling Koch's postulates. Previously, other Botryosphaeriaceae spp. as Diplodia coryli (Guerrero and Pérez 2012) and D. mutila (Moya-Elizondo et al. 2023) have been obtained from canker and dieback of hazelnut in Chile. Recently, N. parvum was reported causing nut rot in hazelnuts in Italy (Wagas et al. 2022). To our knowledge, this is the first report of N. parvum causing canker and branch dieback of hazelnut trees in Chile and worldwide.

Spanish version of the ICIQ-Bowel questionnaire among colorectal cancer patients: construct and criterion validity : Comprehensive assessment of bowel function.

PURPOSE: Bowel complaints are very common among patients with colorectal cancer. However, the most used questionnaires for colorectal cancer survivors do not comprehensively comprise bowel symptoms. This study aimed to examine construct and criterion validity, as well as internal consistency, of the Chilean Version of the International Consultation on Incontinence Questionnaire Bowel Module (ICIQ-B) among people with colorectal cancer. METHODS: Cross-sectional, validation study performed with 106 colorectal cancer patients from Hospital del Salvador, Chile. Bowel function was assessed with the ICIQ-B. Construct validity was assessed with confirmatory factor analysis and hypothesis testing. Specific items of a quality-of-life questionnaire (EORTC QLQ-CR29) were used to correlate with similar ICIQ-B items for criterion validity. For internal consistency, Cronbach's alpha was computed. RESULTS: For construct validity, the confirmatory factor analysis showed that the three factors model did not fit our data. Meanwhile, hypothesis testing favored the construct validity of the instrument, considering that rectal cancer patients showed worse bowel pattern (p = 0.001), bowel control (p = 0.001) and quality of life (p < 0.001) scores compared to colon cancer patients. In addition, those patients assessed before surgery also presented worse scores bowel control (p = 0.023) and quality of life (p = 0.009) compared to post-surgical patients. Regarding criterion validity, the ICIQ-B items showed a significant correlation with similar QLQ-CR29 items. The internal reliability of the instrument was good (Cronbach's α = 0.909). CONCLUSION: Considering that this questionnaire appraises bowel function in more depth, it is recommended for use in clinical practice and research with colorectal cancer patients.

Lutein and Zeaxanthin Content in 21 Plant Species from a Very Humid Premontane Forest in Colombia Palatable for Free-Range Laying Hens.

Xanthophylls, such as lutein and zeaxanthin, have several functions in both plants and humans, including detoxification of oxidants (reactive oxygen species (ROS) and other radicals), maintenance of the structural and functional integrity of biological membranes, and photoprotection from intense light damage. The objective of the present study was to investigate the lutein and zeaxanthin content of 21 species of plants from a very humid premontane forest in Colombia during both dry and rainy seasons. The plants were selected based on being voluntarily eaten by laying hens under free-range conditions. Lutein and zeaxanthin were identified and quantified by high-performance liquid chromatography (HPLC). The results showed that all plants tested contained lutein, at levels ranging from 65.7 to 350 µg/g. Zeaxanthin levels were much lower (2.2 to 26.2 µg/lg) and were detected in only 5 of the 21 plants analyzed. Given that the lutein content of the plants tested in the present study was found to be comparable to that reported in marigold flowers (4-800 µg/g), it is possible that these plants can be used as a source of lutein in free-range laying hen production systems.

Occurrence of Alternaria alternata and A. tenuissima causing black rot in cherry fruits (Prunus avium) in Central Chile.

During the harvest of 2020 and 2021, sweet cherry (Prunus avium) fruit showed a firm rot with irregular pale to dark brown lesions on the fruit surface, with green to light brown fungal growth resembling Alternaria-like infection (Simmons, 2007). Diseased cherries (n= 80 fruit) were collected at harvest in mature (over 10-year-old) commercial orchards of cherry tree varieties Lapins, Regina, Santina, Skeena, and Sweetheart planted in four localities of the regions O´Higgins (33°59´ S, 70°42´W; San Francisco de Mostazal and Graneros) and Maule (35°00'S, 71°23´W; Curicó and Sagrada Familia), Central Chile. The incidence of black rot was 1.9 and 3.2% in O´Higgins and Maule region, respectively, and it was increased to up to 5% during cold storage. The fruit collected previously, were transported to the lab, and surface disinfected in 75% ethanol for 15 s, and rinsed in sterile water. Internal pieces from the junction of diseased and healthy tissues of fruits were placed on potato dextrose agar (PDA, 2%) for 7 days at 20°C. Forty-two isolates of Alternaria-like (Simmons, 2007) were recovered consistently from pure cultures taking hyphal tips from 7 days old cultures. On PDA, 28 isolates (group A) were characterized by cottony, white-gray to green colonies and conidial chains (4 to 10 conidia) with secondary chains (1 to 5 conidia) branching on the conidiophore. Conidia were ovate to obclavate (mean 22.8 ± 5.1 x 8.8 ± 1.5 µm; n=40) with 3 to 7 transepta and 1 longisepta. The remaining 14 isolates (group B) were characterized by cottony, olive-green to olive-brown colonies following a ring pattern of growth and white margins, with conidial chains (4 to 14 conidia) and uncommon secondary chains (1 to 4 conidia). Conidia were obpyriform to ovate, light brown to brown with a cylindrical short beak at the tip (mean 24.7 ± 5.9 × 11.2 ± 1.3 µm; n=40) with 2 to 4 transepta, and 0 to 2 longisepta. Two representative isolates of group A (Sant-02-2020 and Bing-03-2020) and group B (Sant-26-2021 and Skeen-43-2021) were amplified for the Alternaria major allergen (Alt a1), plasma membrane ATPase (ATP), and calmodulin (Cal) loci following the protocols described by Hong et al. (2005) and Lawrence et al. (2013). A MegaBlast search of sequences of group A (GenBank nos. OR267293- OR267294, OR258001- OR258002, and OR267297- OR267298, for Alt a1, ATP, and Cal, respectively) showed 100% similarity to strains UCD10529 and UCD10539 of A. alternata, and group B (GenBank nos. OR267295- OR267296, OR258003- OR258004, and OR258005- OR258006, for Alt a1, ATP, and Cal, respectively) showed 100% similarity to strains EGS 34-015 and A30 of A. tenuissima. Combined phylogenetic analysis using MEGA X clustered isolates Sant-02-2020 and Bing-03-2020, and Sant-26-2021 and Skeen-43-2021 with ex-type of A. alternata and A. tenuissima, respectively. Pathogenicity tests were conducted using isolates of A. alternata (Sant-02-2020; Bing-03-2020) and A. tenuissima (Sant-26-2021; Skeen-43-2021). Detached ripe cherry fruit var. Sweetheart (n=40 fruits/isolate) and Regina (n=40 fruits/isolate) were surfaces disinfested (75% ethanol, 30 s), wounded in the middle with a sterile needle (2 mm in depth), and inoculated with 20 µL of conidial suspension (106 conidia/mL). An equal number of healthy cherries (n=40 fruits) treated with sterile water were used as controls. The experiment was repeated once. All inoculated fruit incubated for 7 days at 22°C, developed between 13 ± 2.7 to 23 ± 2.5 mm and 14.1 ± 1.1 to 19 ± 3.6 mm in lesion diameter for A. alternata and A. tenuissima isolates, respectively. Koch´s postulates were fulfilled by 100% reisolation of the causal pathogen from inoculated fruit, and molecular identification of A. alternata and A. tenuissima isolates. Previously, A. alternata has been described as causing rots on cherries in Chile (Acuña 2010), and China (Zhao and Liu, 2012; Ahmad et al., 2020). To our knowledge, this is the first occurrence of cherry black rot caused by A. alternata and A. tenuissima in Central Chile. Epidemiological studies are necessary to develop integrated management of cherry black rot in Central Chile.

Anti-SpCas9 IgY Polyclonal Antibodies Production for CRISPR Research Use.

The CRISPR/Cas adaptative immune system has been harnessed as an RNA-guided, programmable genome editing tool, allowing for diverse biotechnological applications. The implementation of the system relies on the ability to detect the Cas9 protein in biological samples. This task is facilitated by employing antibodies, which exhibit several advantageous features and applications in the context of tropical neglected diseases. This study reports a one-month immunization scheme with the Cas9 protein fromStreptococcus pyogenes to produce IgY polyclonal antibodies (anti-SpCas9), which can be rapidly isolated by combining yolk de-lipidation with protein salting out using pectin and ammonium sulfate, respectively. Immunodetection assays indicate that the antibodies are highly sensitive, specific, and useful for detecting the SpCas9 protein in promastigotes ofLeishmania braziliensisexpressing exogenous SpCas9. Thus, the simple method for producing anti-SpCas9 IgY antibodies will accelerate CRISPR/Cas-based studies in Leishmania spp. This approach serves as a valuable research tool in this parasite model and holds the potential for wide application in various other biological samples, promoting the implementation of the system. In fact, a bioinformatics approach based on the identification of antigenic determinants in the SpCas9 protein suggests the possibility of using the anti-SpCas9 IgY antibodies in applications such as Prime and Base editing.

Characteristics and outcomes of multisystem inflammatory syndrome in children: A multicenter, retrospective, observational cohort study in Mexico.

Introduction: Multisystem inflammatory syndrome in children associated with coronavirus disease 2019 (MIS-C), a novel hyperinflammatory condition secondary to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, is associated with severe outcomes such as coronary artery aneurysm and death. Methods: This multicenter, retrospective, observational cohort study including eight centers in Mexico, aimed to describe the clinical characteristics and outcomes of patients with MIS-C. Patient data were evaluated using latent class analysis (LCA) to categorize patients into three phenotypes: toxic shock syndrome-like (TSSL)-MIS-C, Kawasaki disease-like (KDL)-MIS-C, and nonspecific MIS-C (NS-MIS-C). Risk factors for adverse outcomes were estimated using multilevel mixed-effects logistic regression. Results: The study included 239 patients with MIS-C, including 61 (26%), 70 (29%), and 108 (45%) patients in the TSSL-MIS-C, KDL-MIS-C, and NS-MIS-C groups, respectively. Fifty-four percent of the patients were admitted to the intensive care unit, and 42%, 78%, and 41% received intravenous immunoglobulin, systemic glucocorticoids, and anticoagulants, respectively. Coronary artery dilatation and aneurysms were found in 5.7% and 13.2% of the patients in whom coronary artery diameter was measured, respectively. Any cause in-hospital mortality was 5.4%. Hospitalization after ten days of symptoms was associated with coronary artery abnormalities (odds ratio [OR] 1.6, 95% confidence interval [CI] 1.2-2.0). Age ≥10 years (OR: 5.6, 95% CI: 1.4-2.04), severe underlying condition (OR: 9.3, 95% CI: 2.8-31.0), platelet count <150,000 /mm3 (OR: 4.2, 95% CI: 1.2-14.7), international normalized ratio >1.2 (OR: 3.8, 95% CI: 1.05-13.9), and serum ferritin concentration >1,500 mg/dl at admission (OR: 52, 95% CI: 5.9-463) were risk factors for death. Discussion: Mortality in patients with MIS-C was higher than reported in other series, probably because of a high rate of cases with serious underlying diseases.

Obtención de la proteína verde fluorescente recombinante y su anticuerpo policlonal Igy

La Proteína Verde Fluorescente (Green Fluorescent Protein, GFP) es ampliamente utilizada en ensayos in vivo e in vitro. Se han generado múltiples variantes de esta proteína para diversificar sus características, como la GFP-enhancer (EGFP) que emite una señal de fluorescencia 35 veces mayor en comparación con la proteína silvestre, siendo implementada como proteína fusión en estudios de localización y estabilidad estructural, entre otros. La detección de esta proteína y sus variantes puede ser directa o indirecta, mediante el uso de anticuerpos anti-GFP. Aunque el uso de GFP es generalizado y de evidente utilidad en investigación y en docencia, los insumos para su estudio exhiben un alto costo dado que deben ser importados, constituyendo un recurso limitado en Colombia. El presente trabajo reporta la clonación y expresión de la proteína recombinante 6xHisEGFP, cuya purificación se completó a partir de la fracción soluble e insoluble del sistema heterólogo Escherichia coli mediante cromatografía de afinidad a metales inmovilizados y electroforesis preparativa, respectivamente. La proteína purificada se implementó como antígeno para la producción de anticuerpos policlonales aviares (IgY) contra la EGFP, los cuales se obtuvieron desde los huevos colectados y el suero de las sangrías de las gallinas inmunizadas. En este sentido, la estrategia metodológica planteada constituye un avance en el desarrollo de un sistema biotecnológico para la producción nacional de herramientas moleculares como los anticuerpos policlonales aviares a bajo costo.

Green Fluorescent Protein (GFP) is widely used in in vivo and in vitro assays. Multiple variants of this protein have been generated to diversify its characteristics, such as the enhancer GFP (EGFP) that emits a 35-fold higher fluorescence signal compared to the wild-type protein, being implemented as a fusion reporter in localization and structural stability studies, among others. Detection of this protein can be direct or indirect, fusing anti-GFP antibodies. Although the use of GFP is generalized and of evident utility in research and teaching, the molecular tools for its study exhibit a high cost since they must be imported, constituting a limited resource in Colombia. This work reports the cloning and expression of the recombinant protein 6xHisEGFP, which purification was completed from the soluble and insoluble fraction of the heterologous Escherichia coli system by immobilized metal affinity chromatography and preparative SDS-PAGE, respectively. The purified protein was implemented as an antigen to produce avian polyclonal antibodies (IgY) against EGFP, which were obtained from collected eggs and blood serum from immunized hens. In this sense, the proposed methodological strategy constitutes an advance in the development of a biotechnological system for the national production of molecular tools such as avian polyclonal antibodies at low-cost.

[Acute coronary syndrome provoked by a coronary intramural hematoma]. / Hematoma intramural coronario como causa de síndrome coronario agudo.

Background: Intramural coronary hematoma (ICH) is an unusual cause of acute coronary syndrome, and it represents a diagnostic challenge, especially in young patients in which it is not considered among the differential causes of acute myocardial ischemia. Clinical case: 40-year-old female, with type 2 diabetes and no other cardiovascular risk factors, who assisted to the Emergency Room with chest pain. In her first evaluation, electrocardiographic abnormalities, and troponin I elevation were found. A cardiac catheterization was performed, in which a proximal obstruction of the left anterior descending artery was observed, and then an optical coherence tomography (OCT) confirmed the presence of an ICH without a dissection flap. A stent was implanted in the obstruction area, with adequate angiographic outcome. The patient had a satisfactory evolution and was discharged to home without evidence of systolic dysfunction and is free of cardiovascular symptoms at 6-month follow-up. Conclusions: ICH must be considered within the differential diagnosis of acute myocardial ischemia in young patients, especially females. Intravascular image diagnosis is essential for the adequate diagnosis and treatment. Treatment must be individualized considering the extent of ischemia.

Introducción: el hematoma intramural coronario (HIC) es una causa poco frecuente de síndrome isquémico coronario agudo y representa un reto diagnóstico, en especial en pacientes jóvenes en los que puede llegar a no ser contemplado dentro de las causas de isquemia cardiaca no ateroesclerótica. Caso clínico: mujer de 40 años, con diabetes tipo 2 y sin otro factor de riesgo cardiovascular, quien acudió al Servicio de Urgencias manifestando dolor torácico. En su evaluación inicial se encontraron alteraciones en el electrocardiograma y elevación de troponina I. Se decidió la realización de coronariografía donde se observó lesión severa de la arteria descendente anterior. El estudio fue complementado con una tomografía de coherencia óptica intracoronaria, la cual confirmó la presencia de HIC sin disección, por lo que se decidió implantar un stent en el segmento arterial afectado con resultado angiográfico exitoso. La convalecencia posterior evolucionó sin complicaciones, lo que posibilitó su egreso a domicilio sin disfunción sistólica en el ecocardiograma y con seguimiento sin eventualidades a los 6 meses. Conclusiones: el HIC es una entidad que debe ser considerada como una causa posible de isquemia miocárdica aguda en pacientes jóvenes y que requiere de la complementación diagnóstica con estudios de imagen intravascular. Su tratamiento debe ser individualizado en el contexto de la extensión de la isquemia.

Inflammatory biomarkers and pendelluft magnitude in ards patients transitioning from controlled to partial support ventilation.

The transition from controlled to partial support ventilation is a challenge in acute respiratory distress syndrome (ARDS) patients due to the risks of patient-self-inflicted lung injury. The magnitude of tidal volume (VT) and intrapulmonary dyssynchrony (pendelluft) are suggested mechanisms of lung injury. We conducted a prospective, observational, physiological study in a tertiary academic intensive care unit. ARDS patients transitioning from controlled to partial support ventilation were included. On these, we evaluated the association between changes in inflammatory biomarkers and esophageal pressure swing (ΔPes), transpulmonary driving pressure (ΔPL), VT, and pendelluft. Pendelluft was defined as the percentage of the tidal volume that moves from the non-dependent to the dependent lung region during inspiration, and its frequency at different thresholds (- 15, - 20 and - 25%) was also registered. Blood concentrations of inflammatory biomarkers (IL-6, IL-8, TNF-α, ANGPT2, RAGE, IL-18, Caspase-1) were measured before (T0) and after 4-h (T4) of partial support ventilation. Pendelluft, ΔPes, ΔPL and VT were recorded. Nine out of twenty-four patients (37.5%) showed a pendelluft mean ≥ 10%. The mean values of ΔPes, ΔPL, and VT were - 8.4 [- 6.7; - 10.2] cmH2O, 15.2 [12.3-16.5] cmH2O and 8.1 [7.3-8.9] m/kg PBW, respectively. Significant associations were observed between the frequency of high-magnitude pendelluft and IL-8, IL-18, and Caspase-1 changes (T0/T4 ratio). These results suggest that the frequency of high magnitude pendelluft may be a potential determinant of inflammatory response related to inspiratory efforts in ARDS patients transitioning to partial support ventilation. Future studies are needed to confirm these results.

Occurrence of leaf rust disease in Oxalis triangularis caused by Puccinia oxalidis in Valdivia, Chile.

False shamrock (Oxalis triangularis), native to South America, is an ornamental and popular plant bulb, commercialized for their attractive shape and color (purple triangular leaves) (Taha et al., 2013). In Chile, a rust was detected in O. triangularis plants growing from April to June in several gardens (n=10) in the city of Valdivia, estimating a disease incidence between 80 and 100%. The symptoms appeared as diffuse chlorotic spots from the upperside of leaves, where infected tissues eventually become completely necrotic, and yellow rust pustules were observed on the underside of leaves. Severe symptoms on infected leaves were consistently observed, showing necrosis on entire leaves. Symptomatic plants (n=50) were collected, and three representative isolates from different localities (OX1, OX2, and OX3) were used for morphological and genetic identification. Uredinia (n=20) were hypophyllous, erumpent, yellow, and irregularly distributed, with sizes from 340 to 850 µm in diameter. Urediniospores (n=150) were yellow, subglobose to globose, equinulate, with measures of 14.2 - 17.7 x 14.7 - 17.2 µm. Teliospores were absent. Based on morphological characters, this rust was identified as a Puccinia sp. These morphological characteristics coincided with those indicated by Safránková (2014), Abbasi et al. (2018), and Khouader et al. (2018). To classify this rust genetically, sequences analyses were performed using the ITS region of the rDNA (ITS4/ITS5) (White et al., 1990). The DNA was extracted using a commercial kit. The results indicated 99% similarity with two reference sequences of P. oxalidis (MH325473 and MH325474) available at GenBank (NCBI http://www.ncbi.nlm.nih.gov/BLAST/). The sequences obtained were deposited in GenBank (ON259085 to ON259087). Based on the maximum parsimony phylogenetic tree, the sequences of Chilean isolates were clustered with those of P. oxalidis references. Pathogenicity tests were conducted using three isolates (OX1 to OX3). Surface disinfection of leaves of O. triangularis (n=36 plants), were performed by spraying 1% NaOCl solution for 1 min. Subsequently, 2 mL of urediniospores suspensions of each isolate (OX1 to OX3) at a concentration of 106 urediniospores/mL, were sprayed with an atomizer on the underside of the leaves of all plants. Urediniospores were obtained following the methodology proposed by Ferrada et al. (2020). Control leaves were disinfested and inoculated with sterile distilled water. Plants of O. triangularis of 90-day-old were incubated in a humid chamber (24°C, 80% HR), with a photoperiod of 12 light /12 dark. At 11 days post-inoculation, all leaves inoculated developed chlorosis spots and pulverulent pustules (averaged 10.9 to 25.4 pustules per leaf), and then at 26 days post-inoculation, affected leaves showed necrotic tissues. The identity of these isolates was confirmed morphologically. The symptoms in the control leaves were negative. To our knowledge, this is the first report of multiple occurrences of the leaf rust disease on gardens of false shamrock caused by P. oxalidis in Valdivia, south of Chile. Previously, P. oxalidis has been reported to cause leaf rust disease in O. triangularis in the Czech Republic (Safránková, 2014) and O. debilis in Korea (Lee et al., 2018). The leaf rust disease could represent a threat to the ornamental gardens of O. triangularis in Valdivia. Currently, epidemiological studies of leaf rust disease are necessary to develop management strategies in gardens of O. triangularis.

Dothiorella sarmentorum Causing Branch Dieback of English Walnut in Maule Region, Chile.

English walnut (Juglans regia), cv. Chandler is the most cultivated tree nut in Chile, with 43,734 ha. In Maule Region, central Chile, English walnut plantings have expanded over an additional 7,000 ha in the last five years. During a routine orchard survey in 2019, branch and twig dieback symptoms were observed in two commercial orchards located in San Rafael (10 years old) and Longaví (12 years old) in the Maule Region, with an incidence of 45% to 65% of affected trees, respectively. Symptomatic branch samples (n = 15) were collected from the two commercial orchards and transported to the laboratory in a cooler and then surface sterilized in 96% ethanol for 3 s and briefly flamed. Cross-section of symptomatic branches revealed brown to dark-brown wedge-shaped wood cankers. Small (5 mm) pieces of wood from the edge of cankered tissues were placed on Potato Dextrose Agar (PDA, 2%) amended with 0.005% tetracycline, 0.01% streptomycin, and 0.1% Igepal CO-630 (PDAm) (Díaz and Latorre 2014) and incubated at 25°C for five days in the dark. Pure cultures were obtained by transferring a hyphal tip from growing colonies to fresh PDA media. Each fungal isolate was recovered from a single diseased branch (47%). Seven isolates (Dsar-1 to Dsar-7) developed dark to olive-brown fast-growing colonies with scarce aerial mycelium after seven days at 25°C on PDA. These isolates showed a dark-olive color on the reverse side of Petri dishes and developed abundant, aggregated, and dark-brown pycnidia after 15 days at 25°C. Conidia were hyaline and aseptate, dark brown, 1-septate, with a brown wall, ovoid with a broadly rounded apex and truncated base, (17.5-) 19.5 ±1.2 (-22.0) x (7.6-) 8.9 ± 0.6 (-10.1) µm (n = 30). These isolates were tentatively identified morphologically as Dothiorella sp. (Phillips et al. 2005). Molecular identification was performed using ITS1/ITS4 and EF1-728F/EF1-986R primers (White et al. 1990; Dissanayake et al. 2015) of the internal transcribed spacer (ITS1-5.8S-ITS2) region and part of the translation elongation factor (EF1-) genes, respectively. A MegaBlast search in GenBank showed a 100% similarity to isolate CBS 115038, the ex-type of Dothiorella sarmentorum. The sequences were added to GenBank (OM161950 to OM161956 for ITS; OM177188 to OM177194 for EF1-). Pathogenicity of two isolates (Dsar-2 and Dsar-7) was tested in the orchard on freshly made pruning wounds on attached branches of 2-year-old-pruned English walnut trees cv. Chandler. A second pathogenicity test was done on freshly made pruning wounds in 1-year-old rooted cuttings (n=15) (40 cm of long) of English walnut cv. Chandler. Each pruning wound was inoculated with 40 µL conidial suspension (105 conidia/mL). Sterile distilled water was used as a control treatment. Both pathogenicity tests were repeated once. After seven months for attached branches and four months for rooted plants, necrotic streaks with a mean length of 81.3 and 44.5 mm were observed below the inoculated pruning wounds, respectively. No necrotic streaks were observed in any of the control wounds. Dothiorella sarmentorum was 100% reisolated from symptomatic tissues of inoculated branches and molecularly identified (EF1-), thus fulfilling Koch's postulates. Recently, D. sarmentorum has been reported causing English walnut dieback in Spain (López-Moral et al. 2020). To our knowledge, this is the first report of D. sarmentorum causing branch dieback of English walnut in Chile. Further studies are needed to know the impact and extent of canker and branch dieback of walnut in commercial orchards in the Maule Region, central Chile.

Concurrent ankle-assisted movement, biofeedback, and proprioceptive stimulation reduces lower limb motor impairment and improves gait in persons with stroke.

BACKGROUND: Persons with stroke live with residual sensorimotor impairments in their lower limbs (LL), which affects their gait. PURPOSE: We investigated whether these residual impairments and resulting gait deficits can be reduced through concurrently applied assisted movement, biofeedback, and proprioceptive stimulation. METHODS: A robotic device provided impairment-oriented training to the affected LL of 24 persons with stroke (PwS) with moderate-to-severe LL impairment. Participants were given 22-30 training sessions over 2-3 months. During training, the interventional device cyclically dorsiflexed and plantarflexed the ankle at 5 deg/s through ±15 deg for 30 min while the participant assisted with the imposed movement. Concurrently, participants received visual biofeedback of assistive joint torque or agonist EMG while mechanical vibration was applied to the currently lengthening (i.e. antagonist) tendon. RESULTS: Sensorimotor impairment significantly decreased over the training period, which was sustained over 3 months, based on the Fugl-Meyer Assessment (FMA-LL) (p < .001), modified Ashworth scale in dorsiflexors (p < .05), and an ankle strength test (dorsiflexors and plantarflexors) (p < .05). Balance and gait also improved, based on the Tinetti Performance Oriented Mobility Assessment (POMA) (p < .05). CONCLUSION: Impairment-oriented training using a robotic device capable of applying assisted movement, biofeedback, and proprioceptive stimulation significantly reduces LL impairment and improves gait in moderately-to-severely impaired PwS.

Escucha activa: base de la relación terapéutica con persona mayor por teleenfermería en contexto pandémico / Active listening: basis of the therapeutic relationship with the elderly person by Telenursing in a pandemic context

El estudio de caso que se presenta a continuación busca resolver el desafío de establecer una relación terapéutica mediante la escucha activa en el contexto remoto de teleenfermería. Para ello se trabajó en un plan de cuidados de enfermería, en base a encuentros remotos con una usuaria de un condominio de viviendas tuteladas por medio de videollamadas, poniendo a prueba la forma en que se realizaba la escucha activa al encontrarse en espacios físicos diferentes. Se realizó una valoración a través de los 11 patrones funcionales de Gordon, priorizando las necesidades detectadas en la usuaria y estableciendo objetivos e intervenciones. Dentro de los resultados obtenidos se destaca que, al existir una óptima metodología de trabajo y retroalimentaciones constantes junto con un buen uso del tiempo, se puede lograr a cabalidad la escucha activa por medio de la teleenfermería a pesar de estar en espacios físicos diferentes. Esto último permite el establecimiento de una relación terapéutica entre el profesional de enfermería y la persona. Se concluye que la teleenfermería es un recurso valioso que es necesario continuar trabajando para que las diferencias entre una atención presencial y una atención remota sean cada vez mínimas[AU]

The case study presented below seeks to resolve the challenge of establishing a therapeutic relationship through active listening in the remote context of tele-nursing. To this end, we worked on a nursing care plan, based on remote meetings with a user of a supervised housing condominium by means of video calls, testing the way in which active listening was carried out when they were in different physical spaces. An assessment was carried out through Gordon's 11 functional patterns, prioritizing the needs detected in the user and establishing objectives and interventions. Among the results obtained, it stands out that, with an optimal work methodology and constant feedback together with a good use of time, active listening can be fully achieved by means of teleconferencing despite the fact that they are in different physical spaces. The latter allows the establishment of a therapeutic relationship between the nurse and the person. It is concluded that tele-nursing is a valuable resource that needs to be further developed so that the differences between face-to-face and remote care become increasingly minimal[AU]

O estudo de caso apresentado abaixo procura resolver o desafio de estabelecer uma relação terapêutica através da escuta activa no contexto remoto da tele-nursão. Para o efeito, trabalhámos num plano de cuidados de enfermagem, baseado em reuniões remotas com um utilizador de um condomínio habitacional supervisionado através de videochamadas, testando a forma como a escuta activa era realizada quando se encontravam em diferentes espaços físicos. Foi feita uma avaliação através dos 11 padrões funcionais de Gordon, dando prioridade às necessidades detectadas no utilizador e estabelecendo objectivos e intervenções. Entre os resultados obtidos, destaca-se que, com uma metodologia de trabalho óptima e um feedback constante juntamente com uma boa utilização do tempo, a escuta activa pode ser plenamente alcançada por meio de teleconferência, apesar de se encontrarem em espaços físicos diferentes. Este último permite o estabelecimento de uma relação terapêutica entre a enfermeira e a pessoa. Conclui-se que a tele-nurmagem é um recurso valioso que precisa de ser mais desenvolvido para que as diferenças entre cuidados presenciais e remotos se tornem cada vez mais mínimas[AU]

Antihyperlipidemic Activity of Gut-Restricted LXR Inverse Agonists.

Hyperlipidemia and increased circulating cholesterol levels are associated with increased cardiovascular disease risk. The liver X receptors (LXRs) are regulators of de novo lipogenesis and cholesterol transport and have been validated as potential therapeutic targets for the treatment of atherosclerosis. However, efforts to develop LXR agonists to reduce cardiovascular diseases have failed due to poor clinical outcomes-associated increased hepatic lipogenesis and elevated low-density lipoprotein (LDL) cholesterol (C). Here, we report that LXR inverse agonists are effective in lowering plasma LDL cholesterol and triglycerides in several models of hyperlipidemia, including the Ldlr null mouse model of atherosclerosis. Mechanistic studies demonstrate that LXR directly regulates the expression of Soat2 enzyme in the intestine, which is directly responsible for the re-uptake or excretion of circulating lipids. Oral administration of a gut-specific LXR inverse agonist leads to reduction of Soat2 expression in the intestine and effectively lowers circulating LDL cholesterol and triglyceride levels without modulating LXR target genes in the periphery. In summary, our studies highlight the therapeutic potential of the gut-restricted molecules to treat hyperlipidemia and atherosclerosis through the intestinal LXR-Soat2 axis.

Emerging Role of Nuclear Receptors for the Treatment of NAFLD and NASH.

Non-alcoholic fatty liver (NAFLD) over the past years has become a metabolic pandemic linked to a collection of metabolic diseases. The nuclear receptors ERRs, REV-ERBs, RORs, FXR, PPARs, and LXR are master regulators of metabolism and liver physiology. The characterization of these nuclear receptors and their biology has promoted the development of synthetic ligands. The possibility of targeting these receptors to treat NAFLD is promising, as several compounds including Cilofexor, thiazolidinediones, and Saroglitazar are currently undergoing clinical trials. This review focuses on the latest development of the pharmacology of these metabolic nuclear receptors and how they may be utilized to treat NAFLD and subsequent comorbidities.

Severe Outbreak of Dry Core Rot in Apple Fruits cv. Fuji Caused by Kalmusia variispora During Pre-harvest in Maule Region, Chile.

Apple (Malus × domestica) is an important fruit crop in Chile, with a cultivated area of 32,313 ha concentrated (63%) in Maule Region (35°25' S). Unusual core rot on 'Fuji' apples was observed at harvest in a commercial orchard in Curicó, Maule Region, with an incidence ranged between 22 to 35% in 2018 and 2019. Previously, in 2017, an incidence of 30% was estimated on 'Fuji' fruits destined to the Asian market. Internal decay symptoms consisted of dry, corky light to dark-brown tissue, within the seed locules initially. In moderate to severe cases, the necrotic lesion progresses deeper into the mesoderm. External symptoms were quite subtle, and typically, the disease goes unnoticed. However, infected fruit ripen earlier. Small pieces (2-3 mm) from the internal lesion margin of symptomatic apples (n = 50) were placed on Potato Dextrose Agar (PDA) (2%) and incubated at 20°C for 10 days. Pure cultures (n = 41) were obtained and transferred to Malt Extract Agar (MEA) (2%). Colonies on MEA produced an even to slight undulating buff margin with white woolly aerial mycelium, and immerse ochreous in the center, changing gray to olivaceous aerial mycelium with age. On the underside, colonies were umber and buff in the center and margin, respectively. After 10 days, numerous densely aggregate dark-brown mature pycnidia were observed. Aseptate conidia were subglobose to cylindrical, straight, and sometimes curved with rounded at both ends, that was initially hyaline to pale olive, thin, smooth wall with mean dimensions of (2.9-) 3.4 (-4.4) x (1.5-) 1.8 (2.2) µm (n=50). Based on morphology, the fungus was identified as Kalmusia variispora (Verkley et al. 2014). The internal transcribed space (ITS), portion of ß-tubulin (TUB), and large subunits of the nuclear ribosomal RNA (LSU) loci were used for molecular identification, using primers ITS4/ITS5, Bt2a/Bt2b, and LR0R/LR5 (Ariyawansa et al., 2014). BLAST searches indicated 100% identity with K. variispora (ex-type CBS 121517). The maximum parsimony phylogenetic analysis placed Chilean isolates in the K. variispora clade. Sequences were deposited in GenBank (OL711706 to OL711709, OL739499 to OL739502 and OL711710 to OL711713 for ITS, TUB and LSU, respectively). Pathogenicity tests were conducted using four K. variispora isolates. 'Fuji' apples (n = 20) were surface disinfested (75% ethanol, 30 s) and then wounded and inoculated with conidial suspension (50 L of 106 conidia/mL) deposited in the middle and into the core region using a sterile fine-tipped micropipette. Additionally, 20 one-year dormant rooted cuttings 'Fuji' and 'Cripps Pink' were pruned and immediately inoculated on the pruning wound. An equal number of apples and rooted cuttings treated with sterile water were used as controls. The experiments were repeated once. All inoculated fruits developed lateral lesions (22 to 37 mm) and dry core rot (18 to 36 mm) symptoms identical to those described in the original outbreak, after 20 days at 20°C in a commercial packing box. The inoculated cuttings produced canker lesions of 10 to 21 mm in length, and dieback symptoms were observed after 3 months. No symptoms were observed on the negative controls. Koch's postulates were fulfilled by 100% reisolating K. variispora. Previously, Alternaria spp. have been reported as the primary pathogen associated with moldy core and dry core rot of apples worldwide (McLeod et al., 2014) and in Chile (Elfar et al., 2018). However, Kalmusia spp. have been associated with dry core rot in apples (McLeod et al., 2014) and have been isolated from canker symptoms on apples in Chile (Díaz et al. 2021). To our knowledge, this is the first report of a severe outbreak of K. variispora causing dry core rot in apples in Chile and worldwide.

Colombian vs. American corn: Quality, chemical composition, mycotoxin content and metabolizable energy

Background: Corn is, quantitatively, one of the most important world crops (ranking second only after wheat) and a key ingredient in animal feeds. Objective: to assess and compare corn quality, mycotoxin content, chemical composition and apparent metabolizable energy (AME) of domestic and imported corn. Methods: Grain quality (USDA grading system) was determined in 30 samples of domestic and 21 samples of imported corn. From each origin, 15 samples were subjected to proximal analysis and 10 were used to determine fatty acid composition. Mycotoxin analysis was conducted on 30 samples of domestic and 23 of imported corn. Results: six of the 30 domestic samples corresponded to US1 grade (highest quality) vs. none of the imported. In the "sample grade" category (lowest quality), 10 and 6 samples corresponded to imported and domestic corn, respectively. Soybeans were found as contaminant in 15 of the 21 imported corn samples. Aspergillus spp. mycotoxins such as ochratoxin A were not detected, and aflatoxins were found in only a few samples at very low levels. Fusariotoxins such as deoxynivalenol and zearalenone were found in 61 and 43% of imported samples, respectively, but in none of the domestic samples. Domestic corn had lower carbohydrate content compared with imported corn (85.4 vs. 86.7%), but higher crude fat (3.8 vs. 3.1%). The AME values for domestic and imported corn were 3,697 and 3,378 kcal/kg, respectively. The fatty acid profiles from both corn types were similar. Conclusion: This study found significant differences between locally-grown and imported corn, particularly in terms of crude fat, AME content, fusariotoxins, and contaminant seeds (soybeans). These findings suggest that locally-grown corn might have nutritional and toxicological advantages over corn imported from the United States.

Antecedentes: El maíz es, cuantitativamente, uno de los cultivos más importantes a nivel mundial (ocupa el segundo lugar después del trigo) y uno de los principales ingredientes en dietas para animales. Objetivo: Evaluar y comparar la calidad del maíz, su contenido de micotoxinas, composición química, y energía metabolizable aparente (AME) del maíz nacional e importado. Métodos: La calidad del grano (sistema de clasificación de la USDA) se determinó en 30 muestras de maíz producido en Colombia y 21 de maíz importado. De cada origen, 15 muestras se sometieron a análisis proximal y 10 se analizaron para determinar el perfil de ácidos grasos. El contenido de micotoxinas se determinó en 30 muestras de maíz nacional y 23 de maíz importado. Resultados: Seis de las treinta muestras nacionales analizadas para calidad de grano correspondieron a grado US1 (la mejor calidad) frente a ninguna de las importadas. En categoría "grado muestra" (la menor calidad) se encontraron 10 y 6 de las muestras de maíz importado y nacional, respectivamente; 15 de las 21 muestras de maíz importado presentaron contaminación con semillas de fríjol soya. No se encontraron micotoxinas de hongos Aspergillus spp. (ocratoxina A), y niveles muy bajos de aflatoxinas en unas pocas muestras. Fusariotoxinas tales como deoxinivalenol y zearalenona se detectaron en el 61 y 43% de las muestras de maíz importado, respectivamente, pero en ninguna muestra de maíz nacional. El maíz nacional presentó menor contenido de carbohidratos (85,4 vs. 86,7%), pero mayor contenido de grasa cruda (3,8 vs. 3,1%) en comparación con el importado, respectivamente. Los valores de AME fueron de 3.697 y 3.378 kcal/kg para el maíz nacional e importado, respectivamente. El perfil de ácidos grasos de ambas procedencias fue similar. Conclusiones: Este estudio encontró diferencias significativas entre el maíz de producción nacional y el importado, en especial en su contenido de grasa cruda y AME, fusariotoxinas, y semillas contaminates (soya). Estos hallazgos sugieren que el maíz de producción nacional puede presentar ventajas de tipo nutricional y toxicológico frente al maíz importado de los Estados Unidos de América.

Antecedentes: O milho é quantitativamente uma das culturas mais importantes em nível mundial (ocupando o segundo lugar depois do trigo), sendo um dos principais ingredientes em dietas para animais. Objetivo: Avaliar e comparar a qualidade do milho, o teor de micotoxinas, a composição química e a energia metabolizável aparente (EMA) do milho nacional e importado. Métodos: A qualidade do grão (sistema de classificação do USDA) foi determinada em 30 amostras de milho produzido na Colômbia e 21 amostras de milho importado. Quinze amostras de cada tipo de milho foram submetidas a análise proximal e dez amostras de cada milho foram analisadas para determinar o perfil de ácidos graxos. O conteúdo de micotoxinas foi determinado em 30 amostras de milho da Colômbia e em 23 amostras de milho importado. Resultados: Seis das trinta amostras colombianas analisadas para qualidade de grão corresponderam a grau US1 (melhor qualidade) frente a nenhuma das amostras importadas. Na categoria "grau amostra" (menor qualidade) foram encontradas 10 e 6 amostras de milhos importados e colombianos, respectivamente; Quinze das 21 amostras de milho importado apresentaram contaminação com sementes de feijão soja. Não foram encontradas micotoxinas de fungos Aspergillus spp. (ocratoxina A) ou apenas níveis muito baixos em poucas amostras (aflatoxinas). As fusariotoxinas, deoxinivalenol e zearalenona foram detectadas em 61 e 43% das amostras de milho importado, respectivamente, mas não foram detectadas em nenhuma amostra do milho colombiano. O milho colombiano apresentou menor conteúdo de carboidratos (85,4 vs. 86,7%) porém maior conteúdo de gordura crua (3,8 vs. 3,1%). Os valores de AME foram 3.697 e 3.378 kcal/kg para as amostras de milho colombiano e importado, respectivamente. O perfil de ácidos graxos de ambos tipos de milho foi similar. Conclusões: Os resultados do presente estudo mostram diferenças significativas entre o milho de origem colombiana e o milho importado dos Estados Unidos, em especial no conteúdo de gordura crua e AME, fusariotoxinas e sementes contaminantes (soja). Estes achados sugerem que o milho de produção colombiana pode apresentar vantagens de tipo nutricional e toxicológica frente ao milho importado dos Estados Unidos.

Duration of the susceptibility of pruning wounds of different ages to infections by Phaeomoniella chlamydospora on grapevine cv. Cabernet Sauvignon in Central Chile.

Grapevine trunk diseases (GTDs) are one of the most important phytosanitary problems that affect grapevines (Vitis vinifera) worldwide. In Chile, Phaeomoniella chlamydospora is the major fungal trunk pathogen associated with GTDs. In the vineyards, the natural infections by P. chlamydospora are associated with air-borne conidia dispersed onto fresh pruning wounds from pycnidia. These pruning wounds are considered an important entrance for fungal trunk pathogens such as P. chlamydospora in the host in the field. However, the duration of the susceptibility of grapevine annual pruning wounds to P. chlamydospora is still unknown in Chile. Therefore, this study aimed to evaluate the period of susceptibility of pruning wounds of different ages to artificial infection of P. chlamydospora on grapevine cv. Cabernet Sauvignon, Central Chile. Artificial inoculations of a conidial suspension (105 conidia/mL) of P. chlamydospora were used to determine the susceptibility of pruning wounds of different ages, from 1, 15, 30, and 45 days after pruning. The experiments were conducted on lignified cuttings in a greenhouse, and on vine spurs in two vineyards (Buin and Nancagua, Central Chile) during two consecutive seasons. The results indicated that the pruning wounds of grapevine cv. Cabernet Sauvignon were very susceptible to infections by P. chlamydospora, with a percentage of pruning wounds infected from 97 to 71% for cuttings, and 96% to 60% for spurs, during the first 15 days after pruning. However, the susceptibility of pruning wounds of different ages in cuttings and spurs of grapevine, generally decreased as the time from pruning to inoculation increased. Moreover, the pruning wounds the pruning wounds remained susceptible to artificial inoculation by P. chlamydospora for up 45 days after pruning with percent of wounds infected from 8.0 to 12.2, and 8.3 to 18.8% on cuttings and spurs of grapevine, respectively. Finally, this study constitutes study constitutes the first research focalized on the susceptibility of pruning wounds of various ages of grapevine cv. Cabernet Sauvignon to artificial inoculations by P. chlamydospora in Central Chile.