Intrauterine infusion of a pathogenic bacterial cocktail is associated with the development of clinical metritis in postpartum multiparous Holstein cows.

Metritis is a uterine disorder common in dairy cattle caused by bacterial infection, with greater incidence in the early postpartum period. The disease causes delayed uterine involution, with a fetid, watery, red-brown discharge, with animals presenting different clinical signs including fever, dullness, inappetence, decreased milk yield, and dehydration. We developed an in vivo model of clinical metritis in Holstein multiparous cows using a pathogenic cocktail of Escherichia coli, Fusobacterium necrophorum, and Trueperella pyogenes. A total of 36 multiparous cows were randomly allocated to 1 of 3 treatment groups of 12 animals each. Cows assigned to the control group received an intrauterine administration of sterile saline solution, those in the low-dose group received a bacterial inoculum containing 106 cfu of Escherichia coli, Trueperella pyogenes, and Fusobacterium necrophorum; and those in the high-dose group received 109 cfu of these same organisms, all within 24 h of parturition. Clinical signs; milk yield; dry matter intake; serum concentration of acute phase proteins, metabolites, and cytokines; and bacterial counts in vaginal discharge were measured from parturition until 14 d in milk. Based on clinical signs, cows in the low-dose group had a greater incidence of metritis (83.3%) compared with the control (9%) or high-dose (25%) group. The low-dose group also had lesser dry matter intake compared with controls and an acute phase protein profile that typified metritis. The low-dose group presented greater relative abundance of bacteria from the genus Fusobacterium in the vaginal discharge compared with control cows, and the high-dose group was similar to the control group. Unexpectedly, intrauterine infusion of cows with a lower dose of bacteria was more effective than a higher dose, indicating that the bacterial load may affect metritis incidence in a nonlinear manner. These findings demonstrate that clinical metritis can be experimentally induced in postpartum multiparous Holstein cows, creating a relevant model for the study of uterine diseases.

Optical fiber thermo-refractometer.

This work presents the implementation of a thermo-refractometer, which integrates the measurement of both refractive index and temperature in a single optical fiber structure. To this purpose, a lossy mode resonance (LMR)-based refractometer is obtained by means of the deposition of a titanium dioxide (TiO2) thin film onto a side-polished (D-shaped) single mode fiber. Measurement and subsequent temperature compensation are achieved by means of a fiber Bragg grating (FBG) inscribed in the core of the D-shaped region. The LMR wavelength shift is monitored in transmission while the FBG (FBG peak at 1533 nm) displacement is observed in reflection. The LMR is sensitive to both the surrounding refractive index (SRI), with a sensitivity of 3725.2 nm/RIU in the 1.3324-1.3479 range, and the temperature (- 0.186 nm/°C); while the FBG is only affected by the temperature (32.6 pm/°C in the 25°C - 45°C range). With these values, it is possible to recover the SRI and temperature variations from the wavelength shifts of the LMR and the FBG, constituting a thermo-refractometer, where it is suppressed the effect of the temperature over the refractometer operation, which could cause errors in the fourth or even third decimal of the measured SRI value.

Proteomic profile of vitrified in vitro-produced bovine embryos (Bos taurus indicus).

BACKGROUND: The proteomic profile of cryopreserved in vitro produced bovine embryos is little known but can provide insights on the successful application of cryo procedures in support of animal breeding. OBJECTIVE: To identify embryonic proteins and biomarkers related to improved cryotolerance of vitrified in vitro produced bovine embryos. MATERIALS AND METHODS: Proteins were isolated from embryo pools (n = 25 embryos per replicate) and analyzed using the nanoLC - MS/MS system. Further, the UniProtKB database (Uniprot -http://www.uniprot.org/) was used for protein identification. Proteins were classified based on their molecular mass, isoelectric point, and enzymatic activity. Post-translational modification predictions and functional gene ontology analysis were performed as well. Finally, a protein-protein interaction network was created to shed light on the embryo interactome. RESULTS: Based on the MS/MS approach, 66 proteins were identified from vitrified Bos taurus embryos. The retrieved proteins were presumably annotated, which allowed a description of the qualitative and functional aspects of the embryo proteome after the vitrification process. CONCLUSION: These findings allowed us to conclude that in vitro-produced vitrified embryos expressed proteins that underlie biological processes related to reproduction, stress and lipid metabolic process, which are essential to maintain embryo viability. doi.org/10.54680/fr22410110512.

Focal adhesion signaling: vascular smooth muscle cell contractility beyond calcium mechanisms.

Smooth muscle cell (SMC) contractility is essential to vessel tone maintenance and blood pressure regulation. In response to vasoconstrictors, calcium-dependent mechanisms promote the activation of the regulatory myosin light chain, leading to increased cytoskeleton tension that favors cell shortening. In contrast, SMC maintain an intrinsic level of a contractile force independent of vasoconstrictor stimulation and sustained SMC contraction beyond the timescale of calcium-dependent mechanisms suggesting the involvement of additional players in the contractile response. Focal adhesions (FAs) are conceivable candidates that may influence SMC contraction. They are required for actin-based traction employed by cells to sense and respond to environmental cues in a process termed mechanotransduction. Depletion of FA proteins impairs SMC contractility, producing arteries that are prone to dissection because of a lack of mechanical stability. Here, we discuss the role of calcium-independent FA signaling mechanisms in SMC contractility. We speculate that FA signaling contributes to the genesis of a variety of SMC phenotypes and discuss the potential implications for mechanical homeostasis in normal and diseased states.

Effect of treatment of pneumonia and otitis media with tildipirosin or florfenicol + flunixin meglumine on health and upper respiratory tract microbiota of preweaned Holstein dairy heifers.

The objective of this randomized clinical study was to compare the effect of 2 antimicrobial interventions, tildipirosin or florfenicol + flunixin meglumine, used for treatment of pneumonia and extralabel treatment for otitis on health parameters and upper respiratory tract (URT) microbiota of preweaned Holstein calves. Housed preweaned Holstein heifers diagnosed with either otitis or pneumonia were assigned into 1 of 2 treatment groups, receiving a single subcutaneous injection of either 4 mg/kg of tildipirosin (TLD; n = 444) or 40 mg/kg of florfenicol combined with 2.2 mg/kg of a nonsteroidal anti-inflammatory, flunixin meglumine (FLF; n = 442). Calves were enrolled and treated on the day of diagnosis of the first case of pneumonia or otitis. If a calf had a recurrent case, the opposite drug was administered, respecting an interval of 5 d between drug injections. Blood samples for leukocyte counts were collected at 0, 2, 4, and 6 d after treatment, and rectal temperature was measured daily during the 5 d after treatment. Ear scores were observed from calves with otitis. Additionally, swabs of the URT were collected from a subset of 20 calves in each treatment group at d 0, 3, 6, 9, and 11 following enrollment for analysis of URT microbiota through next-generation sequencing of the 16S rRNA gene and quantitative PCR. Swabs were also collected from a comparative group of 20 healthy calves that did not receive any drug. No differences were observed between groups for recurrence risk of either pneumonia (TLD = 32.4%; FLF = 29.7%) or otitis (TLD = 72.7%; FLF = 73.6%). Similarly, no differences were observed for the total number of treatments for pneumonia (TLD = 1.45; FLF = 1.42) or otitis (TLD = 2.96; FLF = 3.07). On the other hand, both drugs reduced rectal temperature, ear scores, and leukocyte counts, with FLF calves having a greater reduction in rectal temperature within 4 d after treatment. Both TLD and FLF reduced the total bacterial load when compared with healthy untreated calves, but no differences were observed between treatment groups. Furthermore, compared with the untreated group, treated calves had lower mean relative abundances (MRA) of the genera Mannheimia, Moraxella, and Pasteurella within 11, 9, and 3 d after treatment, respectively; however, no significant differences were observed between TLD and FLF. On the other hand, MRA of Mycoplasma was not decreased by both treatments compared to untreated animals, and a higher MRA was observed in the TLD group during 11 d after treatment in comparison to FLF and untreated calves. Based on this data, we concluded that both drugs used in the study were effective in reducing rectal temperature, ear scores, leukocyte counts, and MRA of the genera Mannheimia, Pasteurella, and Moraxella in the URT, and calves treated with FLF had a greater reduction in rectal temperature.

Immunization with a novel recombinant protein (YidR) reduced the risk of clinical mastitis caused by Klebsiella spp. and decreased milk losses and culling risk after Escherichia coli infections.

The primary objective of this study was to evaluate the protective efficacy of a novel recombinant subunit vaccine containing the protein YidR (rYidR) against clinical mastitis (CM) caused by Klebsiella spp. and Escherichia coli. Given that E. coli infection is known to cause metritis, we also evaluated the effect of rYidR vaccination on the incidence of metritis and conception at the first artificial insemination. Retained placenta and abortion incidence, milk production and composition, and serological responses to specific antigens were also evaluated. In total, 3,107 cows were blocked by parity and randomly allocated into 1 of 3 treatment groups: experimental recombinant subunit vaccine containing the YidR protein (rYidR); commercial vaccine composed of Klebsiella pneumoniae siderophore receptors and porin protein (Kleb-SRP; KlebVax, Epitopix, Willmar, MN); and sterile water adjuvanted with aluminum hydroxide (20%; placebo). Vaccinations were performed at the dry-off for cows, and at 223 ± 3 d of pregnancy for pre-fresh heifers. A second administration was given at 21 ± 3 d after the first injection. Vaccination with rYidR significantly reduced the incidence of CM caused by Klebsiella spp. (3.2%) when compared with the placebo (5.1%) group. No difference was observed on risk of Klebsiella CM between Kleb-SRP (5.9%) and placebo groups. Cows in the rYidR group that experienced E. coli CM had a lower risk of death or culling (12.5%) compared with the Kleb-SRP (27.6%) and placebo groups (27.8%). Furthermore, among cows that developed E. coli CM, rYidR-immunized cows produced more milk than did cows in the placebo and Kleb-SRP groups. Regardless of CM occurrence, rYidR-immunized cows tended to have higher milk production up to the eighth month of lactation than cows in the other groups. No significant effect of treatment was observed on the overall incidence of abortion and metritis; however, the risk of retained placenta tended to be lower for the rYidR group (4.7%) compared with the placebo group (6.7%). In addition, primiparous cows in the rYidR group had the highest conception risk at the first artificial insemination (48.3%) compared with the placebo (39.5%) group, and no significant difference was observed when the Kleb-SRP (40.1%) group was compared with the placebo group. Generally, higher antibody serum titers (IgM and IgG) were observed for the immunized groups compared with the placebo. In conclusion, the rYidR vaccine reduced the risk of CM caused by Klebsiella spp. and the mortality or culling of cows with E. coli infections. Other benefits of the novel vaccine include maintenance of milk production after CM caused by E. coli, and higher conception risk at the first service in primiparous cows compared with cows in the placebo and Kleb-SRP groups.

Negatively controlled, randomized clinical trial comparing different antimicrobial interventions for treatment of clinical mastitis caused by gram-positive pathogens.

The general objective of this study was to evaluate the effect of 3 intramammary antibiotic interventions using 2 commercially available antibiotics with narrow- or broad-spectrum activity on cure rates of clinical mastitis (CM) caused by gram-positive bacteria. We also compared the efficacy of treatment protocols, including a negative control, on outcomes at the cow and mammary quarter level. Before the onset of the study, 5,987 animals more than 12 mo old were randomly preassigned to 1 of 4 protocols in the event of gram-positive CM (except for Staphylococcus aureus and Trueperella pyogenes) during lactation: 3 infusions with 62.5 mg of amoxicillin performed 12 h apart (AMOX-L); 5 infusions once a day with 62.5 mg of amoxicillin (AMOX-EL); 5 infusions once a day with 125 mg of ceftiofur hydrochloride (CEFT-L); or negative control, no treatment performed until 5 d after diagnosis (NEG-CTR). Randomization was performed to preassign 90% of cows to one of the antibiotic protocols (30% in each group) and 10% to the negative control. A total of 696 quarter cases of CM met the inclusion criteria and were evaluated in the study. Quarter-level outcomes were assessed based on 5 milk samples collected up to 14 ± 3 d following enrollment (i.e., first day of treatment), whereas variables at the cow level [composite somatic cell count (SCC), milk production, and survival in the herd] were assessed up to 90 d after CM diagnosis. Streptococcus uberis, followed by Streptococcus dysgalactiae, were the main causes of gram-positive CM. Overall, clinical cure was higher for CEFT-L than for AMOX-EL, and no difference was observed between CEFT-L and AMOX-L. Likewise, no significant differences were detected on overall bacteriological cure, although some treatment effects were observed at the species level. Compared with antibiotic-treated groups, quarters assigned to NEG-CTR had higher counts of colony-forming units (cfu), 16S rRNA gene copy numbers, and Streptococcus relative abundance (RA) until d 5 after enrollment. Quarters treated with AMOX-L had higher cfu counts on d 5, 8, and 14 after enrollment compared with the other antibiotic protocols. In addition, the RA of Streptococcus spp. was higher on d 14 after enrollment for AMOX-treated quarters compared with the CEFT-L group. Linear score of SCC was higher for AMOX-treated cows than for CEFT-L in the first test day after CM. However, cows assigned to AMOX-L had higher milk production than those submitted to the AMOX-EL and CEFT-L protocols. In conclusion, the 2-d protocol with 3 intramammary infusions of amoxicillin (narrow-spectrum antimicrobial) had similar overall clinical and bacteriological cures as 5 administrations (once a day) with ceftiofur hydrochloride (wide spectrum). No significant difference was observed on CM recurrence and cow survival. However, quarters treated with 5-d protocols were more effective at reducing milk cfu counts than quarters in the AMOX-L protocol. In addition, lower Streptococcus spp. RA was observed in ceftiofur-treated quarters compared with the amoxicillin protocols at d 14 after CM diagnosis. Based on results of microbiome and bacterial load (quantitative PCR and cfu count) up to 5 d after CM diagnosis, antibiotic use remains an indispensable strategy for treatment of CM caused by gram-positive bacteria.

Evaluation of the sensitivity of the Express View function in the Mirocam® capsule endoscopy software.

Background: A new computer algorithm called Express-View has recently been introduced by Mirocam, but data concerning its application and efficacy are scarce.Objective: To evaluate the lesion detection rate, per-patient sensitivity and the diagnostic accuracy using Express-View.Methods: All patients who performed CE between January 2018 and June 2019, whose indication was obscure gastrointestinal bleeding (OGIB) and with findings on CE, were included. Lesions identified in conventional reading were selected and considered as reference.Results: Eighty-nine patients were included, 50.6% male, with a mean age of 68.4 years-old (±12.3). The Express-View mode detected 85.5% of lesions previously detected by conventional reading (524 out of 613). There were 89 missed lesions, mainly erosions or ulcers (44.9%) and angioectasias (38.2%). The lesion detection rate was found to be lower in the jejunum and ileum compared to extra-small bowel locations and duodenum (p = .04). Although Express-View had a per-patient sensitivity for all lesions of 56.2% and a per-patient sensitivity for all clinically significant lesions of 83.1%, it achieved a diagnostic accuracy of 91%.Conclusions: The per-patient sensitivity for all lesions was shown to be below expectations, although the lesion detection rate, the per-patient sensitivity for all clinically significant lesions, and the diagnostic accuracy were shown to be higher.

Recombinant protein subunit vaccine reduces puerperal metritis incidence and modulates the genital tract microbiome.

The objective of this study was to evaluate the efficacy of 3 vaccine formulations containing proteins (FimH, leukotoxin, and pyolysin), inactivated whole cells (Escherichia coli, Fusobacterium necrophorum, and Trueperella pyogenes), or both, in the prevention of postpartum uterine diseases. A randomized clinical trial was conducted at a commercial dairy farm; 800 heifers were assigned into 1 of 4 different treatment groups: control, vaccine 1 (bacterin and subunit proteins), vaccine 2 (bacterin), and vaccine 3 (recombinant subunit proteins), and each heifer received a subcutaneous injection of its respective treatment at 240 ± 3 and 270 ± 3 d of gestation. Vaccination significantly reduced the incidence of puerperal metritis when compared with control (9.1% vs. 14.9%, respectively; odds ratio 0.51). Additionally, vaccine 3 was found to reduce the incidence of puerperal metritis when compared with the control (8.0% vs. 14.9%, respectively; odds ratio 0.46). Reproduction was improved for metritic cows that were vaccinated, and the effect was stronger for cows that were treated with vaccine 3. In general, vaccination decreased the total vaginal bacterial load and decreased the vaginal load of F. necrophorum by 9 d in milk. Vaccination reduced the prevalence of puerperal metritis in the first lactation of dairy cows, leading to less metritic disease and improved reproduction.

Liquid Storage of Geophagus brasiliensis semen in the Presence of Different Extenders.

BACKGROUND: In order to preserve the genetic diversity of cichlid fish in gene banks, it is necessary to use certain extenders to maintain the integrity of spermatozoa cells during cooling. OBJECTIVE: To evaluate the effects of different extenders on the quality parameters of cooled semen of Geophagus brasiliensis. MATERIALS AND METHODS: Semen samples were collected from seven adult fish and diluted with five extenders: Beltsville Thawing Solution (BTS™), Hanks' Balanced Salt Solution (HBSS), Tris-glucose, Ginsburg's Fish Ringers, and Phosphate buffered Saline. All parameters were evaluated in fresh semen samples and after cooling at 4°C at 0, 24, 48, and 96 h to evaluate cell viability (membrane integrity, DNA, and mitochondrial functionality) and motility rate and weather motility. RESULTS: The BTS and Tris-glucose resulted in the best outcomes (P<0.05) in terms of membrane integrity assessments (35.1% and 30.9%, respectively), DNA integrity (71.6%; 75.7%), mitochondrial function (26.9%; 28.0%) and motility rate (8.6%; 8.6%), respectively, for semen cooled to 4°C for 96 h. However, the 48-h period motility after cooling in BTS was superior to all other treatments. CONCLUSION: BTS and Tris-glucose can be considered as the best extenders for the cold storage of Geophagus brasiliensis spermatozoa.

Cryopreservation of Bovine Semen Using Extract of Bertholletia excelsa (Brazil Nuts).

BACKGROUND: Semen cryopreservation is essential in animal breeding programs for improving the availability of genetic resources from animals with high breeding value. OBJECTIVE: To evaluate the addition of Brazil nut extract as a replacement for egg yolk in bovine semen cryopreservation. MATERIALS AND METHODS: Semen was collected from five Nelore bulls and cryopreserved with the addition (treatments) of 0, 25, 50, 75, or 100% Brazil nut extract in the cryoprotectant medium. After thawing, spermatic cells were evaluated for morphology, plasma membrane integrity, spermatic kinetics, and in vitro fertilization. The experimental design was in randomized blocks, and the data were submitted to regression analysis. RESULTS: The minor-type and total defects, and plasma membrane integrity were affected (P < 0.05) as a function of egg yolk substitution with Brazil nut extract. There was a significant effect (P < 0.05) of Brazil nut extract addition on the spermatic kinetics and cleavage rate. CONCLUSION: The addition of Brazil nut extract in the cryoprotective medium as a substitute of egg yolk for freezing bovine semen negatively affects sperm quality and fertility.

Three-Dimensional Correlative Imaging of a Malaria-Infected Cell with a Hard X-ray Nanoprobe.

Benefiting from the recent advances of synchrotron X-ray nanoprobes, we demonstrate three-dimensional (3D) correlative nanoimaging on malaria-infected human red blood cells. By combining X-ray fluorescence tomography and phase contrast nanotomography on the same cell with sub-100 nm pixel size, we establish a routine workflow from the data acquisition, data processing, to tomographic reconstruction. We quantitatively compare the elemental volumes obtained with different reconstruction methods, with the total variation minimization giving the most satisfactory results. We reveal elemental correlations in different cell compartments more reliably on reconstructions as opposed to 2D projections. Finally, we determine for the first time the 3D mass fraction maps of multiple elements at the subcellular level. The estimated total number of Fe atoms and the total mass of red blood cells show very good agreement with previously reported values.

In Vitro and In Vivo Efficiency of Extenders Added to Thawed Stallion Semen.

BACKGROUND: Addition of extenders to thawed semen could improve fertility. OBJECTIVE: To determine the efficiency of extenders to increase viability of thawed semen, measured by sperm parameters in vitro and pregnancy rates after artificial insemination (AI). MATERIALS AND METHODS: Sperm motility and acrosin activity were measured during a thermoresistance test (TRT). RESULTS: Progressive motility decreased (P<0.05) after 30 min in thawing semen treated with saline solution (SS) and only after 60 min with Tyrode's solution (TS) or freezing diluent (FD). The total motility decreased (P<0.05) after 60 min in thawed semen treated with SS, and after 90 min in thawed semen containing TS or FD. The acrosin activity decreased (P<0.05) after 60 min during the TRT, but there was no difference among treatments throughout the TRT. The pregnancy rates were similar among thawed-semen supplemented with SS, TS or FD. CONCLUSION: The extenders neither improve sperm parameters nor enhance AI results.

The Addition of AçaÍ Palm Fruit Extract in the Cryopreservation Diluent of Bull's Semen with Low Performance on Freezing.

BACKGROUND: Semen freezing is of great importance for animal production because it allows the use and the rapid diffusion of the genetic material from economically important animals. OBJECTIVE: To evaluate the effect of açai (Euterpe oleracea; Arecaceae family) extract addition to the semen cryopreservation diluent on the morphology, sperm motility parameters, and plasma membrane integrity of spermatozoa. MATERIALS AND METHODS: The ejaculates, obtained from five bulls with low performance on semen freezing, were fractionated and distributed according to the experimental group. The control samples did not have açaí extract added, whereas to the treated groups were added 5, 10, 15 or 20 mg ml-1 of açaí extract into the semen diluent. The sperm morphology was evaluated with a formalin-saline-buffered solution. The plasma membrane integrity was evaluated by the epifluorescent test, while the cellular kinetics was assessed by an automated analysis of the spermatic movement. RESULTS: The sperm defects showed a linearly decreasing effect (P < 0.05) with the addition of different concentrations of açaí extract. The plasma membrane integrity was higher (P < 0.05) after the açaí addition to the cryopreservation diluent. There was no significant effect (P > 0.05) of the açaí extract on the kinetics of spermatozoa. CONCLUSION: The addition of açaí extract to the cryopreservation diluent provided better preservation of the structural integrity of the sperm plasma membrane in the bull's semen with low tolerance to the cryopreservation process.

Quantitative region-of-interest tomography using variable field of view.

In X-ray computed tomography, the task of imaging only a local region of interest (ROI) inside a larger sample is very important. However, without a priori information, this ROI cannot be exactly reconstructed using only the image data limited to the ROI. We propose here an approach of region-of-interest tomography, which reconstructs a ROI within an object from projections of different fields of view acquired on a specific angular sampling scheme in the same tomographic experiment. We present a stable procedure that not only yields high-quality images of the ROI but keeps as well the quantitative contrast on the reconstructed images. In addition, we analyze the minimum number of projections required for ROI tomography from the point of view of the band region of the Radon transform, which confirms this number must be estimated based on the size of the entire object and not only on the size of the ROI.

Electromyographic characterisation of abdominal wall trigger points developed after caesarean section and response to local anaesthesia: an observational study.

OBJECTIVE: This study examines the electromyography pattern of abdominal trigger points developed after a caesarean section, and the association between clinical response and local anaesthetic injection. DESIGN: Prospective cohort study. SETTING: A tertiary university hospital. POPULATION: Twenty-nine women with chronic pelvic pain associated with trigger points after a caesarean section were included in the study. METHODS: Participants received needle electromyography before treatment, then underwent a treatment protocol consisting of trigger-point injection of 2 ml of 1% lidocaine. The protocol was repeated once a week for 4 weeks. The clinical responses of the patients were compared 1 week after and 3 months after treatment. The clinical trial is registered with the Brazilian Clinical Trials Registry (REBEC) under RBR-42c6gz (www.ensaiosclinicos.gov.br/rg/RBR-42c6gz/). MAIN OUTCOME MEASURES: Needle electromyography and algometry results and pain reduction. RESULTS: Fifteen patients had abnormal electromyography findings; 14 had normal findings. The rates of response 1 week and 3 months after treatment within the abnormal electromyography group were 95 and 87%, respectively. In the normal group, the rate was 38% both 1 week after and 3 months after treatment. CONCLUSIONS: Trigger points developed after caesarean section, even without clinical symptoms or signs of neuralgia, may originate from neuropathies. Electromyographic abnormalities were associated with pain remission after anaesthesia injection; normal electromyography findings were associated with undiagnosed causes of pain, such as adhesions. TWEETABLE ABSTRACT: Trigger points developed after caesarean section are neuropathies, even in the absence of classical neuralgia.

Assistive devices: an effective strategy in non-pharmacological treatment for hand osteoarthritis-randomized clinical trial.

This study aims to evaluate the use of assistive devices as a strategy in non-pharmacological treatment for hand osteoarthritis (HOA). This is a randomized, prospective, parallel, assessor-blinded clinical trial, in which patients with a diagnosis of HOA were randomly allocated to an intervention group (IG), where they received assistive devices for daily life activities, or to a control group (CG), where they received a guideline leaflet with information on joint protection and disease features. The primary outcomes considered were occupational performance, measured by the Canadian Occupational Performance Measure (COPM), and hand function was evaluated through the Score for the Assessment and Quantification of Chronic Rheumatoid Affections of the Hands (SACRAH). The secondary outcomes were pain, measured by the visual analog scale (VAS), and quality of life, measured by the World Health Organization Quality of Life Instrument, Short Form (WHOQOL-BREF). We compared both outcomes before and after interventions and outcomes between groups. Participants from the two groups were assessed at the time of inclusion in the study, 30, and 90 days after initial evaluation. Out of the 39 patients included, 19 were allocated to the IG and 20 to the CG. Only two patients from the CG did not complete the follow-up period. The patients' hand function and occupational performance improved after intervention (30 days-SACRAH-p < 0.05; COPM-p < 0.05; VAS-p < 0.05). When comparing results between the groups, there was a statistical difference in COPM (performance-p < 0.001; and satisfaction-p < 0.001), in the first reevaluation carried out. The use of assistive devices has proved to be an effective alternative in non-pharmacological treatment for HOA. CLINICAL TRIAL REGISTRATION: NCT02667145.

Temporal expression of pluripotency-associated transcription factors in sheep and cattle preimplantation embryos.

SummaryPluripotency-associated transcription factors (PATFs) modulate gene expression during early mammalian embryogenesis. Despite a strong understanding of PATFs during mouse embryogenesis, limited progress has been made in ruminants. This work aimed to describe the temporal expression of eight PATFs during both sheep and cattle preimplantation development. Transcript availability of PATFs was evaluated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in eggs, cleavage-stage embryos, morulae, and blastocysts. Transcripts of five genes were detected in all developmental stages of both species (KLF5, OCT4, RONIN, ZFP281, and ZFX). Furthermore, CMYC was detected in all cattle samples but was found from cleavage-stage onwards in sheep. In contrast, NR0B1 was detected in all sheep samples but was not detected in cattle morulae. GLIS1 displayed the most significant variation in temporal expression between species, as this PATF was only detected in cattle eggs and sheep cleavage-stage embryos and blastocysts. In silico analysis suggested that cattle and sheep PATFs share similar size, isometric point and molecular weight. A phenetic analysis showed two patterns of PATF clustering between cattle and sheep, among several mammalian species. In conclusion, the temporal expression of pluripotency-associated transcription factors differs between sheep and cattle, suggesting species-specific regulation during preimplantation development.

Variable Inter-assay Estimation of Sperm DNA Fragmentation In Stallions Classified as Good and Bad Semen Freezers.

BACKGROUND: Semen cryopreservation causes DNA damage, thus requiring continuous monitoring. OBJECTIVE: To compare two assays for sperm DNA fragmentation (SDF) from stallions with contrasting semen freezability. MATERIALS AND METHODS: Thirteen stallions were classified as good semen freezers (GSF) or bad semen freezers (BSF). Ejaculates were cryopreserved with three diluents. Semen was subject to SDF evaluation using the sperm chromatin structure assay (SCSA) and Halomax after thawing (0 h) and after a 4 h thermoresistance test. RESULTS: On semen of BSF, analysis by SCSA was similar between evaluations, but Halomax showed increased SDF at 4 h. The GSF group was similar between time points in both assays. Diluents did not affect SDF, irrespective of the assay. Halomax showed differences for BSF between time points, differently from SCSA. Linear regression did not show any correlation between assays. CONCLUSION: The use of Halomax should be encouraged for sperm DNA fragmentation analysis in horse frozen-thawed semen, particularly under field conditions.

Theoretical modulation of singlet/triplet chemiexcitation of chemiluminescent imidazopyrazinone dioxetanone via C8-substitution.

Coelenterazine, a member of the imidazopyrazinone class of chemiluminescent substrates, presents significant potential as a dynamic probe of reactive oxygen species in a biological environment, such as a superoxide anion, in which these species are important in cellular biology and pathology. The objective of the current study was to understand in what way the efficiency of singlet and triplet chemiexcitation could be modulated, towards a more efficient use of imidazopyrazinone-based compounds as dynamic chemiluminescent probes. To this end the thermolysis of imidazopyrazinone dioxetanone, substituted at the C8-position with electron-donating or electron-withdrawing groups, was characterized with a theoretical approach based on density functional theory. Substituents with different electron-donating/withdrawing characters have only a limited effect on the singlet chemiexcitation of anionic dioxetanone. For neutral dioxetanone, both electron-withdrawing and weak electron-donating substituents increase singlet chemiexcitation, to the contrary of strong electron-donating groups. During their thermolysis reaction, all molecules presented regions of degeneracy with triplet states, thereby indicating the possibility of triplet chemiexcitation.