Analysis and applications of respiratory surface EMG: report of a round table meeting.

Surface electromyography (sEMG) can be used to measure the electrical activity of the respiratory muscles. The possible applications of sEMG span from patients suffering from acute respiratory failure to patients receiving chronic home mechanical ventilation, to evaluate muscle function, titrate ventilatory support and guide treatment. However, sEMG is mainly used as a monitoring tool for research and its use in clinical practice is still limited-in part due to a lack of standardization and transparent reporting. During this round table meeting, recommendations on data acquisition, processing, interpretation, and potential clinical applications of respiratory sEMG were discussed. This paper informs the clinical researcher interested in respiratory muscle monitoring about the current state of the art on sEMG, knowledge gaps and potential future applications for patients with respiratory failure.

Carcass, meat quality traits, and economic analysis of Nellore bulls fed with finishing feedlot diets containing mechanically processed corn silage.

Effects of mechanical processing (MP) of corn silage and its inclusion in feedlot diets on carcass and meat quality traits of Nellore (Bos indicus) were analyzed. Seventy-two bulls aged approximately 18 months and with an initial average body weight of 392.8 ± 22.3 kg were used. The experimental design was a 2 × 2 factorial arrangement, considering the concentrate-roughage (C:R) ratio (40:60 or 20:80), MP of silage and their interactions. After slaughter, hot carcass weight (HCW), pH, temperature, backfat thickness (BFT), and ribeye area (REA), yields of meat cuts (tenderloin, striploin, ribeye steak, neck steak, and sirloin cap), meat quality traits and economic analysis were evaluated. A lower final pH was found in the carcasses of animals consuming diets containing MP versus unprocessed silage (pH = 5.81 versus 5.93). Carcass variables (HCW, BFT, and REA) and meat cut yields were not affected by treatments. The C:R 20:80 increased the intramuscular fat (IMF) content by approximately 1%, without affecting moisture, ash, and protein contents. Meat/fat color (L*, a* and b*) and Warner-Bratzler shear force (WBSF) were similar among treatments. The results indicated that the MP of corn silage in finishing diets can provide better carcass pH results in Nellore bulls, without negatively influencing carcass weight, fatness, and meat tenderness (WBSF). The IMF content of meat was slightly improved using a C:R 20:80 and lower total costs per arroba produced (3.5%), daily costs per animal/day (4.2%), and cost per ton of feeds (5.15%) were found with MP silage.

Using rumination and activity data for early detection of anaplasmosis disease in dairy heifer calves.

Bovine anaplasmosis causes considerable economic losses in dairy cattle production systems worldwide, ranging from $300 million to $900 million annually. It is commonly detected through rectal temperature, blood smear microscopy, and packed cell volume (PCV). Such methodologies are laborious, costly, and difficult to systematically implement in large-scale operations. The objectives of this study were to evaluate (1) rumination and activity data collected by Hr-Tag sensors (SCR Engineers Ltd.) in heifer calves exposed to anaplasmosis; and (2) the predictive ability of recurrent neural networks in early identification of anaplasmosis. Additionally, we aimed to investigate the effect of time series length before disease diagnosis (5, 7, 10, or 12 consecutive days) on the predictive performance of recurrent neural networks, and how early anaplasmosis disease can be detected in dairy calves (5, 3, and 1 d in advance). Twenty-three heifer calves aged 119 ± 15 (mean ± SD) d and weighing 148 ± 20 kg of body weight were challenged with 2 × 107 erythrocytes infected with UFMG1 strain (GenBank no. EU676176) isolated from Anaplasma marginale. After inoculation, animals were monitored daily by assessing PCV. The lowest PCV value (14 ± 1.8%) and the finding of rickettsia on blood smears were used as a criterion to classify an animal as sick (d 0). Rumination and activity data were collected continuously and automatically at 2-h intervals, using SCR Heatime Hr-Tag collars. Two time series were built including last sequence of -5, -7, -10, or -12 d preceding d 0 or a sequence of 5, 7, 10, or 12 d randomly selected in a window from -50 to -15 d before d 0 to ensure a sequence of days in which PCV was considered normal (32 ± 2.4%). Long short-term memory was used as a predictive approach, and a leave-one-animal-out cross-validation (LOAOCV) was used to assess prediction quality. Anaplasmosis disease reduced 34 and 11% of rumination and activity, respectively. The accuracy, sensitivity, and specificity of long short-term memory in detecting anaplasmosis ranged from 87 to 98%, 83 to 100%, and 83 to 100%, respectively, using rumination data. For activity data, the accuracy, sensitivity, and specificity varied from 70 to 98%, 61 to 100%, and 74 to 100%, respectively. Predictive performance did not improve when combining rumination and activity. The use of longer time-series did not improve the performance of models to predict anaplasmosis. The accuracy and sensitivity in predicting anaplasmosis up to 3 d before clinical diagnosis (d 0) were greater than 80%, confirming the possibility for early identification of anaplasmosis disease. These findings indicate the great potential of wearable sensors in early identification of anaplasmosis diseases. This could positively affect the profitability of dairy farmers and animal welfare.

Investigation of the protective suitability of a dental fluorinated varnish by means of X Ray fluorescence and Raman spectroscopy.

BACKGROUND AND AIM: Evaluating the protective effect in human enamel of a fluorinated varnish after enduring a citric acid erosive challenge. METHODS: An in vitro model was developed considering the intraoral environment, human saliva and acid erosive procedures. The evaluation of the enamel specimens was undertaken through the direct analysis of enamel by means of Raman spectroscopy and Energy Dispersive X Ray Fluorescence (EDXRF). Ten tooth specimens per group were analysed during three stages: 1- before treatment; 2- After varnish (treatment group) or toothpaste (control) application; 3- After citric acid cycle. Additionally, Particle Induced Gamma Ray emission (PIGE) was used to gauge the fluorine uptake by enamel after the application of the varnish (stage 2). Results were presented as mean and standard deviation with ANOVA and Tukey post hoc performed considering a significance level of 0.05. RESULTS: A significant (p < 0.05) higher Ca levels were detected in treatment group at stage 2 (37.4 ± 0.4 w/w%) and 3 (37.1 ± 0.1) when compared to the control group. After varnish application in treatment group, depolarization ratios were significant lower (p < 0.05) and anisotropy were significant higher (p < 0.05), however no differences were detected in FWHM. CONCLUSIONS: The use of a fluorinated dental varnish suggests a protective effect for human enamel against dental erosion demineralization process which was detectable in an in vitro model.

Kidney injury molecule-1 and urinary gamma-glutamyl transferase as biomarkers of acute kidney injury in cats.

OBJECTIVES: To evaluate the concentration of kidney injury molecule-1 and activity of urinary gamma-glutamyl transferase in cats with urethral obstruction and healthy cats. MATERIALS AND METHODS: Blood and urine samples were collected from a group of 15 healthy cats (control group) and a group of 20 cats with urethral obstruction at presentation, and 24 hours and 7 days after unblocking the obstruction. The serum creatinine, urinary creatinine and urinary gamma-glutamyl transferase were measured by spectrophotometry and kidney injury molecule-1 by the sandwich enzyme-linked immunosorbent assay. RESULTS: On presentation, cats with obstruction had serum creatinine concentration and urinary gamma-glutamyl transferase index higher than healthy cats (mean difference 544 µmol/L, 95% confidence intervals 222 to 865 µmol/L, and 0.0022 U/µmol-uCre, 0.00043 to 0.0039 U/µmol-uCre, respectively), urine creatinine concentration lower (mean difference 25,624 µmol/L, 17,329 to 33,919 µmol/L), and no significant difference in the kidney injury molecule-1/urinary creatinine ratio (mean difference 13 pg/µmol-uCre, -33 to 59 pg/µmol-uCre). In the group of cats with urinary obstruction, over time serum creatinine decreased, urine creatinine increased, urinary gamma-glutamyl transferase index did not change significantly, and kidney injury molecule-1/urinary creatinine ratio increased. CLINICAL SIGNIFICANCE: Cats with post-renal obstruction and potential intrinsic renal damage had higher urinary gamma-glutamyl transferase index than healthy cats at the time of presentation and showed increase in kidney injury molecule-1/urinary creatinine ratio over time.

Genomics and transcriptomics insights into luteolin effects on the beta-rhizobial strain Cupriavidus necator UYPR2.512.

Cupriavidus necator UYPR2.512 is a rhizobial strain that belongs to the Beta-subclass of proteobacteria, able to establish successful symbiosis with Mimosoid legumes. The initial steps of rhizobium-legumes symbioses involve the reciprocal recognition by chemical signals, being luteolin one of the molecules involved. However, there is a lack of information on the effect of luteolin in beta-rhizobia. In this work, we used long-read sequencing to complete the genome of UYPR2.512 providing evidence for the existence of four closed circular replicons. We used an RNA-Seq approach to analyse the response of UYPR2.512 to luteolin. One hundred and forty-five genes were differentially expressed, with similar numbers of downregulated and upregulated genes. Most repressed genes were mapped to the main chromosome, while the upregulated genes were overrepresented among pCne512e, containing the symbiotic genes. Induced genes included the nod operon and genes implicated in exopolysaccharides and flagellar biosynthesis. We identified many genes involved in iron, copper and other heavy metals metabolism. Among repressed genes, we identified genes involved in basal carbon and nitrogen metabolism. Our results suggest that in response to luteolin, C. necator strain UYPR2.512 reshapes its metabolism in order to be prepared for the forthcoming symbiotic interaction.

Groundwater Quality in the Yucatan Peninsula: Insights from Stable Isotope and Metals Analysis.

High surface water-groundwater connectivity characterizes watersheds underlain by karsts, increasing contaminant transport risks. However, karsts are highly complex, making research necessary to understand the transport of contaminants from the surface, through the aquifer, to discharge areas. In Yucatan, the lack of waste water treatment raises the risk of groundwater contamination. We monitored stable isotopes (δ18 O-NO3 and δ15 N-NO3 ), cadmium, and lead to document waste water contamination and transport during the rainy and dry seasons, using water samples collected along the Ring of Cenotes during each season. Specific conductance and pH showed no consistent seasonality, with conductance ranging from 0.5 to 55 mS/cm and pH ranging from 6.6 to 8.6 for most samples. Nitrate concentrations in the cenotes averaged 205 ± 260 µM and no seasonal pattern was observed. Cd and Pb concentrations were 0.1 to 37.9 µg/L and 0.2 to 243.2 µg/L, respectively. Nitrate stable isotope values were 2.6 to 27.2‰ for δ18 O and 1.2 to 20.7‰ for δ15 N. The statistical relationship between δ15 N and δ18 O, in dry season samples, indicated that denitrification was occurring. A scale measure for waste water recognition showed: (1) high variability among sites probably related with dry/rainy seasons and/or diverse anthropogenic activities; and (2) specific water quality variables that contribute to contamination at each site during each season. Importantly, our analyses indicate that in the area surrounding the Ring of Cenotes, waste water exhibits spatial and temporal patterns related to complex transport and dilution processes, as is the case in karsts in general.

Identifying and managing patient–ventilator asynchrony: An international survey / Identificación y manejo de la asincronía paciente-ventilador: encuesta internacional

Objective To describe the main factors associated with proper recognition and management of patient–ventilator asynchrony (PVA). Design An analytical cross-sectional study was carried out. Setting An international study conducted in 20 countries through an online survey. Participants Physicians, respiratory therapists, nurses and physiotherapists currently working in the Intensive Care Unit (ICU). Main variables of interest Univariate and multivariate logistic regression models were used to establish associations between all variables (profession, training in mechanical ventilation, type of training program, years of experience and ICU characteristics) and the ability of HCPs to correctly identify and manage 6 PVA. Results A total of 431 healthcare professionals answered a validated survey. The main factors associated to proper recognition of PVA were: specific training program in mechanical ventilation (MV) (OR 2.27; 95%CI 1.14–4.52; p=0.019), courses with more than 100h completed (OR 2.28; 95%CI 1.29–4.03; p=0.005), and the number of ICU beds (OR 1.037; 95%CI 1.01–1.06; p=0.005). The main factor influencing the management of PVA was the correct recognition of 6 PVAs (OR 118.98; 95%CI 35.25–401.58; p<0.001). Conclusion Identifying and managing PVA using ventilator waveform analysis is influenced by many factors, including specific training programs in MV, the number of ICU beds, and the number of recognized PVAs. (AU)

Objetivo Describir los factores asociados al correcto reconocimiento y manejo de la asincronía paciente-ventilador (APV). Diseño Estudio analítico transversal. Ámbito Estudio internacional realizado en 20 países mediante una encuesta a través de Internet. Participantes Médicos, terapeutas respiratorios, enfermeras/os y fisioterapeutas que trabajan actualmente en unidades de cuidados intensivos (UCI). Principales variables de interés Se utilizó un análisis uni y multivariado para describir la asociación entre todas las variables (profesión, formación en ventilación mecánica, tipo de programa de formación, años de experiencia y características de la UCI en la cual trabajan los profesionales) con la correcta identificación y manejo de 6 APV. Resultados Un total de 431 profesionales respondieron una encuesta validada previamente. Los factores asociados a una correcta identificación de 6 APV fueron: haber completado un programa de formación específico sobre ventilación mecánica (OR: 2,27; IC 95%: 1,14-4,52; p=0,019), programa de formación con más de 100h (OR: 2,28; IC 95%: 1,29-4,03; p=0,005) y el número de camas de UCI (OR: 1,037; IC 95%: 1,01-1,06; p=0,005). El principal factor asociado a un adecuado manejo de la APV fue la correcta identificación de 6 APV (OR: 118,98; IC 95%: 35,25-401,58; p<0,001). Conclusiones La identificación y el manejo de la asincronía paciente-ventilador, mediante el análisis de las curvas del ventilador está influenciada por programas de formación, específicos sobre ventilación mecánica, el número de camas de la UCI y el número de asincronías identificadas. (AU)

Monitoring bovine tick fever on a dairy farm: An economic proposal for rational use of medications.

In this study, we evaluated the monitoring of tick fever (TF) in a Brazilian dairy farm in the Minas Gerais state, Brazil, from July 10 to August 4, 2018. We aimed to identify diagnostic and treatment flaws in the protocol adopted by the farm, and to establish a novel and accurate TF monitoring protocol based on precision dairy farming and rational use of antimicrobials and antiparasitic drugs, while evaluating the economic benefits of the proposed strategy. We monitored TF in 395 heifer calves aged between 3 and 14 mo. According to the farm's standard protocol, all calves with an increase of 0.5°C in rectal temperature compared with the previous week's measurement were treated for Anaplasma spp. and Babesia spp. Blood smears were collected from the tail tip of the treated calves. During the last week of the study, we prepared blood smears of all calves regardless of treatment indication. Economic analysis was performed. The results indicated that at least 56.86% (261/459) of the calves did not require treatment for TF, whereas only 23.09% (106/459) had treatment indications. Negative blood smears (45.97%; 211/459) indicated the possibility of calves being affected by another disease or a condition that was not being adequately treated or those not necessarily sick. These results demonstrate the excessive use of medications, representing a direct economic loss, in addition to potentially favoring the occurrence of resistance to antimicrobials. In contrast, 9.42% (26/276) of calves had no treatment indication based on rectal temperature but had treatment indications based on blood smears. Only 5.73% (42/735) of blood smears had co-infection with hemopathogens, and none had triple co-infection. Therefore, we proposed the monitoring of TF using rectal temperature and microscopic analysis. If implemented, this strategy would result in a direct annual savings of approximately $22,638.96 (77.99%) related to medication for the treatment of TF. Therefore, implementing the proposed protocol would be cheaper than treatment based only on rectal temperatures. The currently implemented TF protocols overestimate the occurrence of TF, resulting in overtreatment. Thus, implementing a TF monitoring protocol based on a microscopy tool is justified, with benefits including rational use of medication, potential to generate savings, and reduced morbidity and mortality rates, in addition to enabling other diagnoses.

Identifying and managing patient-ventilator asynchrony: An international survey.

OBJECTIVE: To describe the main factors associated with proper recognition and management of patient-ventilator asynchrony (PVA). DESIGN: An analytical cross-sectional study was carried out. SETTING: An international study conducted in 20 countries through an online survey. PARTICIPANTS: Physicians, respiratory therapists, nurses and physiotherapists currently working in the Intensive Care Unit (ICU). MAIN VARIABLES OF INTEREST: Univariate and multivariate logistic regression models were used to establish associations between all variables (profession, training in mechanical ventilation, type of training program, years of experience and ICU characteristics) and the ability of HCPs to correctly identify and manage 6 PVA. RESULTS: A total of 431 healthcare professionals answered a validated survey. The main factors associated to proper recognition of PVA were: specific training program in mechanical ventilation (MV) (OR 2.27; 95%CI 1.14-4.52; p=0.019), courses with more than 100h completed (OR 2.28; 95%CI 1.29-4.03; p=0.005), and the number of ICU beds (OR 1.037; 95%CI 1.01-1.06; p=0.005). The main factor influencing the management of PVA was the correct recognition of 6 PVAs (OR 118.98; 95%CI 35.25-401.58; p<0.001). CONCLUSION: Identifying and managing PVA using ventilator waveform analysis is influenced by many factors, including specific training programs in MV, the number of ICU beds, and the number of recognized PVAs.

Effect of kernel processing and particle size of whole-plant corn silage with vitreous endosperm on dairy cow performance.

Kernel processing and theoretical length of cut (TLOC) of whole-plant corn silage (WPCS) can affect feed intake, digestibility, and performance of dairy cows. The objective of this study was to evaluate for lactating dairy cows the effects of kernel processing and TLOC of WPCS with vitreous endosperm. The treatments were a pull-type forage harvester without kernel processor set for a 6-mm TLOC (PT6) and a self-propelled forage harvester with kernel processor set for a 6-mm TLOC (SP6), 12-mm TLOC (SP12), and 18-mm TLOC (SP18). Processing scores of the WPCS were 32.1% (PT6), 53.9% (SP6), 49.0% (SP12), and 40.1% (SP18). Twenty-four Holstein cows (139 ± 63 d in milk) were blocked and assigned to six 4 × 4 Latin squares with 24-d periods (18 d of adaptation). Diets were formulated to contain 48.5% WPCS, 15.5% citrus pulp, 15.0% dry ground corn, 9.5% soybean meal, 6.8% low rumen degradability soybean meal, 1.8% calcium soap of palm fatty acids (FA), 1.7% mineral and vitamin mix, and 1% urea (dry matter basis). Nutrient composition of the diets (% of dry matter) was 16.5% crude protein, 28.9% neutral detergent fiber, and 25.4% starch. Three orthogonal contrasts were used to compare treatments: effect of kernel processing (PT6 vs. SP6) and effect of TLOC (particle size; SP6 vs. SP12 and SP12 vs. SP18). Cows fed SP6 produced 1.2 kg/d greater milk yield with no changes in dry matter intake, resulting in greater feed efficiency compared with PT6. Cows fed SP6 also produced more milk protein (+36 g/d), lactose (+61 g/d), and total solids (+94 g/d) than cows fed PT6. The mechanism for increased yield of milk and milk components involved greater kernel fragmentation, starch digestibility, and glucose availability for lactose synthesis by the mammary gland. However, cows fed SP6 had lower chewing time and tended to have greater levels of serum amyloid A compared with PT6. Milk yield was similar for SP6 and SP12, but SP12 cows tended to have less serum amyloid A with greater chewing time. Cows fed SP18 had lower total-tract starch digestibility and tended to have lower plasma glucose and produce less milk compared with cows fed SP12. Compared with PT6, feeding SP6 raised linear odd-chain FA concentration in milk. Similarly, a reduction of these same FA occurred for SP12 compared with SP6. Cows fed SP6 had greater proportion of milk C14:1 and C16:1 compared with PT6 and SP12. Lesser trans C18:1 followed by greater C18:0 concentrations were observed for SP12 and PT6 compared with SP6, which is an indication of more complete biohydrogenation in the rumen. Under the conditions of this study, the use of a self-propelled forage harvester with kernel processing set for a 12-mm TLOC is recommended for WPCS from hybrids with vitreous endosperm.

Seven years of evaluation of ectoparasites and vector-borne pathogens among ring-tailed coatis in an urban park in southeastern Brazil.

Wild animals have been recognized as potential reservoirs of vector-borne pathogens. Proximity between these animals and urban areas increases the need to know which pathogens these are and whether they can infect domestic animals and humans. In Mangabeiras Municipal Park in Belo Horizonte, Brazil, coatis live near the urban area, which is mainly occupied by human residents and their domestic animals. Therefore, the objective of this study was to detect, through molecular and direct methods, the presence of ectoparasites and hemoparasites in coatis. A total of 216 samples were collected, of which 209 samples were from first-captures and seven were from recaptures. The following parasites were found: ticks of the genus Amblyomma, lice of the species Neotrichodectes pallidus and fleas of the species Rhopalopsyllus lutzi lutzi and Ctenocephalides felis felis. All the samples were negative for the family Anaplasmataceae and the species Leishmania sp. and Trypanosoma cruzi. The hemoparasites Trypanosoma evansi, Hepatozoon procyonis, Babesia sp. and Sarcocystis neurona were found. The area of the present study is not endemic for T. evansi, which therefore suggests that these coatis may be acting as reservoirs or sentinels of this parasite. This finding is of great epidemiological importance and should be investigated more closely. Thus, this study showed that there is a great variety of pathogens in the park that transit among coatis and, probably, among other animals that inhabit or live close to the park.

Type I interferon receptors and interferon-τ-stimulated genes in peripheral blood mononuclear cells and polymorphonuclear leucocytes during early pregnancy in beef heifers.

This study characterised the expression of interferon (IFN)-τ-stimulated genes (ISGs) and Type I IFN receptors in circulating polymorphonuclear cells (PMNs) of beef heifers and compared it with expression in peripheral blood mononuclear cells (PBMCs) up to Day 20 of gestation. Nelore heifers (n=26) were subjected to fixed-time AI (FTAI) on Day 0. PMNs and PBMCs were isolated on Days 0, 10, 14, 16, 18 and 20 after FTAI. The abundance of target transcripts (ubiquitin-like protein (ISG15), 2'-5'-oligoadenylate synthetase 1 (OAS1), myxovirus resistance 1 (MX1), myxovirus resistance 2 (MX2), IFN receptor I (IFNAR1) and IFN receptor 2 (IFNAR2)) was determined using real-time quantitative polymerase chain reaction and compared between pregnant (n=8) and non-pregnant (n=9) females. In both PBMCs and PMNs, ISG15 and OAS1 expression was greater in pregnant than non-pregnant heifers on Days 18 and 20. There were no significant differences in the expression of ISGs between PBMCs and PMNs. A time effect on expression was found for IFNAR1 in PBMCs and IFNAR2 in PMNs, with decreased expression of both genes on Days 18 and 20. When the expression of these genes was compared between cell types only in pregnant heifers, IFNAR2 expression in PMNs had an earlier decrease when compared to its expression in PBMCs, starting from Day 18. In conclusion, PMNs do not respond earlier to the conceptus stimulus, and ISG15 and OAS1 expression in both PMNs and PBMCs can be used as a suitable marker for pregnancy diagnosis on Days 18 and 20. In addition, gestational status did not affect IFNAR1 and IFNAR2 expression, but IFNAR2 showed a distinct response between PMNs and PBMCs of pregnant heifers.

Evaluation of the effect of fluorinated tooth bleaching products using polarized Raman microscopy and particle induced gamma-ray emission.

In this in vitro study, the effect of the application of tooth bleaching products in human enamel was evaluated using polarized Raman microscopy, particle induced gamma-ray emission (PIGE) and Vickers Hardness test. Due to their acidic nature, teeth whitening products are associated with changes in enamel mineralization. Consequently, products have appeared in the market that promote the incorporation of fluorine in order to decrease the solubility of the hydroxyapatite in enamel and prevent demineralization. This way, four commercial products with different active principle concentrations: 16% carbamide peroxide (Opalescence PF® and VivaStyle®) or 6% hydrogen peroxide (Opalescence Go PF® and VivaStyle Paint On®) and presence or not of fluorine were compared. The information on the crystalline state of the enamel was provided by the determination of the depolarization ratio of the symmetric stretching band of phosphate (at 959 cm-1). Furthermore, the content and uptake of F was evaluated using PIGE in the two fluorinated products as well as in one negative control group. In order to evaluate the microhardness of enamel by means of Vickers test, another group of polished samples was prepared (using Opalescence PF®) and evaluated. Conversely to what could be expected, the obtained results determined a statistically significant decrease of depolarization ratio, leading to an increase of mineralization after the application of the bleaching products, except for VivaStyle®. For this group, no significant variation was obtained before-after treatment, most likely due to the acidity of the product (pH = 5.8). Additionally, an increase of concentration of F in the dental tissues was determined for the fluorinated products. On the other hand, enamel polishing, required for the application of the Vickers test, led to increased susceptibility to erosion, resulting in decreased hardness and an increased enamel depolarization ratio.

Chemiluminescent ELISA with multi-epitope proteins to improve the diagnosis of canine visceral leishmaniasis.

Diagnosing canine visceral leishmaniasis (CVL) is difficult because clinical signs of the disease are non-specific and a many infected animals in endemic areas, as in Brazil, are asymptomatic. Serological tests are the most common diagnostic methods employed, but most have limitations. For this reason, the implementation of a rapid, sensitive, and specific diagnostic test for CVL has become increasingly important. In this study, we adapted a chemiluminescent enzyme-linked immunosorbent assay (CL ELISA), using two multi-epitope recombinant proteins (PQ10 and PQ20) and a crude Leishmania antigen produced using promastigotes of L. infantum, as antigens to detect CVL infection in animals from Belo Horizonte. To investigate cross-reactions, samples from dogs with other infections (babesiosis, ehrlichiosis and Trypanosoma cruzi) were tested. Assay performance validations were conducted to analyse parameters such as variability, reproducibility, and stability. CL ELISA sensitivity/specificity with PQ10 antigen was 93.1%/80.0%; with the PQ20 protein 93.1%/96.6%; and with the crude antigen 75%/73.3%. Inter-assay variability and inter-operator coefficient of variation were <7% and <15%, with PQ10 and PQ20, respectively. The accuracy of the CL ELISA was classified as excellent for PQ10 (AUC = 0.95) and PQ20 (AUC = 0.98) and moderate for the crude antigen (AUC = 0.77). The kappa score for qualitative agreement between two plate lots was excellent for PQ10 (0.89) and good for PQ20 (0.65). PQ20 remained more stable than PQ10. The CL ELISA with recombinant proteins is a promising tool to diagnose CVL.

Bothrops alternatus Snake Venom Induces Cytokine Expression and Oxidative Stress on Renal Function.

BACKGROUND: Envenomation caused by Bothrops alternatus is common in Southern Brazil. Acute Kidney Injury occurs after Bothrops snakebite and more information is necessaryrequired to understand its mechanism. OBJECTIVE: The objective was to evaluate the effect of Bothrops alternatus venom (BaV) on renal cells and rat isolated kidney function. METHODS: Wistar rats (n = 6, weighing 260-320 g) were perfused with a Krebs-Henseleit solution containing 6 g 100 mL-1 of bovine serum albumin. After 30 minutes, the kidneys were perfused with BaV to a final concentration of 1 and 3 µgmL-1; and subsequently were evaluated for Perfusion Pressure (PP), Renal Vascular Resistance (RVR), Urinary Flow (UF), Glomerular Filtration Rate (GFR), and percentage of electrolyte tubular transport. Renal histological analysis, cytokine release, oxidative stress and cytotoxicity in renal proximal tubular cells were assessed. RESULTS: BaV reduced PP, RVR, GFR, UF, total and proximal sodium transport (%TNa+), and chloride (%TCl-) in the isolated kidney perfusion model. Histological analysis of perfused kidneys disclosed the presence of proteinaceous material in the glomeruli and renal tubules, vacuolar tubular epithelial cell degeneration, Bowman's capsule degeneration, swelling of glomerular epithelial cells, glomerular atrophy and degeneration, and the presence of intratubular protein. Cytokine release (TNF-α, IL-1ß, IL-10) and oxidative stress were increased in the kidneys. The viability of LLC-MK2 cells (IC50: 221.3 µg/mL) was decreased by BaV and necrosis was involved in cell death. CONCLUSION: These findings indicate that BaV modifies functional parameters in an isolated perfused kidney model and has cytotoxic effects on renal lineage cells.

The effect of length of storage and sodium benzoate on the nutritive value of reconstituted sorghum grain silages for dairy cows.

Twenty Holstein cows at 168 ± 87 d in milk (mean ± SD) were assigned to a 4 × 4 Latin square design with a 2 × 2 factorial arrangement to evaluate the effects of 2 storage lengths (30 or 90 d) and the presence of sodium benzoate (control or 0.2% as fed) on the nutritive value of reconstituted sorghum grain silages (RSGS). For each treatment, dry ground sorghum grain was rehydrated to 35% moisture and ensiled in 200-L plastic drums. The treatments were RSGS stored for 30 d without sodium benzoate (30 CON), RSGS stored for 30 d with sodium benzoate (30 BEN), RSGS stored for 90 d without sodium benzoate (90 CON) and RSGS stored for 90 d with sodium benzoate (90 BEN). Diets contained 16.3% RSGS. Silages stored for 90 d had higher concentrations of 1,2-propanediol, soluble protein, and ammonia nitrogen than did those stored for 30 d. Sodium benzoate reduced ethanol and ethyl-ester formation. Silages stored for 90 d had higher starch (89.3 vs. 86.9%) and protein (57.1 vs. 54.0%) digestibility compared with silages stored for 30 d. The ruminal acetate-to-propionate ratio tended to be lower in RSGS stored for 90 d than in RSGS stored for 30 d (3.75 vs. 3.34). Milk yield increased from 30.0 kg/d in cows fed RSGS stored for 30 d to 31.2 kg/d in cows fed RSGS stored for 90 d, without a change in dry matter intake (23.5 kg/d on average). Hence, feed efficiency and milk N efficiency also had tendencies to increase in cows fed RSGS stored for 90 d. Sodium benzoate did not alter cow performance but slightly increased plasma glucose (65.2 vs. 63.6 mg/dL). In conclusion, increasing the storage period of RSGS from 30 to 90 d improved starch and protein digestibility, milk yield, and feed efficiency.

High ammonium supply impairs photosynthetic efficiency in rice exposed to excess light.

Mechanisms involving ammonium toxicity, excess light, and photosynthesis are scarcely known in plants. We tested the hypothesis that high NH4+ supply in presence of high light decreases photosynthetic efficiency of rice plants, an allegedly tolerant species. Mature rice plants were previously supplied with 10 mM NH4+ or 10 mM NO3- and subsequently exposed to 400 µmol m-2 s-1 (moderate light-ML) or 2000 µmol m-2 s-1 (high light-HL) for 8 h. HL greatly stimulated NH4+ accumulation in roots and in a minor extent in leaves. These plants displayed significant delay in D1 protein recovery in the dark, compared to nitrate-supplied plants. These responses were related to reduction of both PSII and PSI quantum efficiencies and induction of non-photochemical quenching. These changes were also associated with higher limitation in the donor side and lower restriction in the acceptor side of PSI. This later response was closely related to prominent decrease in stomatal conductance and net CO2 assimilation that could have strongly affected the energy balance in chloroplast, favoring ATP accumulation and NPQ induction. In parallel, NH4+ induced a strong increase in the electron flux to photorespiration and, inversely, it decreased the flux to Rubisco carboxylation. Overall, ammonium supply negatively interacts with excess light, possibly by enhancing ammonium transport towards leaves, causing negative effects on some photosynthetic steps. We propose that high ammonium supply to rice combined with excess light is capable to induce strong delay in D1 protein turnover and restriction in stomatal conductance, which might have contributed to generalized disturbances on photosynthetic efficiency.

Role of extracellular vesicles during oocyte maturation and early embryo development.

The follicle is a dynamic microenvironment in the ovary where the oocyte develops. Intercellular communication between somatic cells and the oocyte inside the follicle is essential to generate a competent gamete. Extracellular vesicles are nanoparticles secreted by cells that mediate cell-to-cell communication in the follicle microenvironment and can be obtained from the follicular fluid. These extracellular vesicles have been studied as biomarkers and supplementation tools to mimic physiological conditions during assisted reproductive techniques because they are vehicles of bioactive molecules. Therefore, this paper reviews the importance of changes in the ovarian follicle and the effects of extracellular vesicles from follicular fluid during oocyte maturation and early embryo development. Finally, we propose that is important to consider the source of the extracellular vesicles to improve diagnostic methods and to increase invitro embryo production.