Relationship among thermal environment, stage of lactation, body characteristics, yield and milk constituents of dairy Gyr cows managed on pasture.

Our aims were to evaluate changes in body characteristics, milk yield and milk constituents as well as to determine the relationship between the thermal environment and production characteristics during the first lactation of dairy Gyr cows managed on pasture. Between 2013 and 2015, forty-five primiparous dairy Gyr cows were evaluated from prepartum to 10 months of lactation in Southeast of Brazil. Body weight, body condition score (BCS), subcutaneous fat thickness (SFT), milk yield (305 d), and milk constituents were collected monthly and progesterone was collected weekly. Additionally, we determined the temperature humidity index (THI) based on microclimate data. Overall, the cows lost body weight until six months of lactation and there was a progressive decrease in BCS, SFT, milk yield and milk lactose as the months in lactation progressed. In contrast, there was an increase in milk fat, milk protein and milk solids. The thermal environment did not pose a consistent heat challenge, nevertheless, we found a positive correlation between the average THI two days before milk collection with milk yield, fat and lactose contents, but in contrast a negative correlation was found with total solids and protein. In conclusion, the THI and months of lactation affected the yield and constituents of milk. However, more studies are necessary to understand the impacts of body characteristics and thermal environment on yield and milk constituents throughout the productive life of Gyr dairy cows.

Comparative effectiveness of peroneus longus tendon (PLT) autografts versus hamstring tendon (HT) autografts in anterior cruciate ligament reconstruction: a comprehensive systematic review and meta analysis.

The hamstring tendon (HT) autograft is currently the most widely utilised autograft option for anterior cruciate ligament (ACL) reconstruction. However, recent studies endorse the peroneus longus tendon (PLT) autograft as a viable alternative. To evaluate this, we systematically reviewed randomised controlled trials (RCTs) to compare the efficacy of PLT against HT autografts. Our search encompassed Cochrane, Embase, OVID, PubMed, and Scopus databases for RCTs comparing outcomes of PLT and HT autografts in ACL reconstruction. Primary outcomes included Lysholm and International Knee Documentation Committee (IKDC) scores, while secondary outcomes involved American Orthopaedic Foot and Ankle Society (AOFAS) scores, graft diameters and donor-site complications. Statistical analysis was performed using Review Manager 5.4 (Cochrane Collaboration) and heterogeneity was assessed with I2 statistics. 683 patients from 6 RCTs were included, with 338 (49.5%) patients treated with PLT autografts. Follow-up ranged from 12 to 30 months. Despite lower preoperative Lysholm scores in the PLT group, no significant differences were observed at 6 and 12 months. Although preoperative and 6-month IKDC scores were lower in the PLT group, no significant differences were found at 12 and 24 months. AOFAS scores showed no significant preoperative difference, but slightly lower scores were noted in the PLT group at 12 or 24 months. There was no significant difference in graft diameter, while donor-site complications were fewer in the PLT group. In summary, the PLT autograft is a promising and non-inferior alternative to the HT autograft, demonstrating equivalent outcomes in patient-reported knee and ankle metrics, comparable graft diameters and fewer donor-site complications.

Expanding the genetic spectrum of ALKU syndrome: Compound heterozygosity for two deleterious variants in SMG8 gene.

Alzahrani-Kuwahara syndrome (ALKUS) is a neurodevelopmental disorder that includes microcephaly, facial dysmorphism, and variable congenital and eye malformations. We present the first case of ALKUS described in the European population caused by two variants in compound heterozygosity of the gene SMG8. We present a patient with two variants in compound heterozygosity in the SMG8 gene identified by in trio whole exome sequencing based in next generation sequencing (xGEN® Exome Research Panel, Nextseq550 platform). International case reporting (CARE) criteria were followed. Patient written consent was obtained through legal responsible persons. We describe a 27-year-old male, the second child of a healthy and non-consanguineous couple, whose genetic analysis showed two variants in compound heterozygosity, c.1159C > T (p.Arg387*) and c.2407del (p.Arg803Glyfs*10), in the SMG8 gene, both classified as likely pathogenic. As described by Fatema Alzahrani et al. in a series of eight patients, our patient had global developmental delay with impaired intellectual development, facial dysmorphism, and limb disproportion. Additionally, our patient had lower limb spastic paraparesis, marked osteotendinous hyperreflexia with extensor plantar response bilaterally and paretic gait. Our patient resembles the phenotype described by Fatema Alzahrani et al., however, he is the first patient with two SMG8 deleterious variants in compound heterozygosity, and the first to exhibit pyramidal signs and gait disorder as part of the phenotype.

Association of PTX3 gene polymorphisms and PTX3 plasma levels with leprosy susceptibility.

BACKGROUND: Pentraxin 3 (PTX3) is a soluble pattern recognition receptor that plays a crucial role in modulating the inflammatory response and activating the complement system. Additionally, plasma PTX3 has emerged as a potential biomarker for various infectious diseases. The aim of this study was to evaluate the association of PTX3 gene polymorphisms and PTX3 plasma levels with susceptibility to leprosy and clinical characteristics. METHODS: Patients with leprosy from a hyperendemic area in the Northeast Region of Brazil were included. Healthy household contacts and healthy blood donors from the same geographical area were recruited as a control group. The rs1840680 and rs2305619 polymorphisms of PTX3 were determined by real-time PCR. Plasma levels of PTX3 were determined by ELISA. RESULTS: A total of 512 individuals were included. Of these, 273 were patients diagnosed with leprosy; 53 were household contacts, and 186 were healthy blood donors. No association was observed between PTX3 polymorphisms and susceptibility to leprosy or development of leprosy reaction or physical disability. On the other hand, plasma levels of PTX3 were significantly higher in patients with leprosy when compared to household contacts (p = 0.003) or blood donors (p = 0.04). It was also observed that PTX3 levels drop significantly after multidrug therapy (p < 0.0001). CONCLUSIONS: Our results suggest that PTX3 may play an important role in the pathogenesis of leprosy and point to the potential use of this molecule as an infection marker.

Protonated and Sodiated Cyclophosphamide Fragmentation Pathways Evaluation by Infrared Multiple Photon Dissociation Spectroscopy.

Cyclophosphamide (CP or CTX) is a widely used antineoplastic agent, and the evaluation of its efficacy and its impacts on the environment are dependent on tandem mass spectrometry (MSn) techniques. Because there is no dedicated experimental study to characterize the actual molecular nature of the CP fragments upon collision-induced dissociation, this work evaluated the chemical structure of the fragments of protonated and sodiated CP and CP protonation sites by infrared multiple photon dissociation spectroscopy supported by density functional theory calculations. This study allowed us to propose a new fragment structure and confirm the nature of multiple fragments, including those relevant for transitions used for CP quantitative and qualitative analyses. Our results also show that there is no spectroscopic evidence that can rule out the existence of aziridinium fragments, making it clear that further studies on the nature of iminium/aziridinium fragments in the gas phase are necessary.

Implementation and Evaluation of a Prior Authorization Workflow for New-Start Inpatient Medications in Preparation for Discharge.

Purpose: Medications that require prior authorization can complicate the discharge planning process. This study implemented and evaluated a process for identifying and completing prior authorizations during the inpatient setting prior to patient discharge. Methods: A patient identification tool was developed within the electronic health record to alert the patient care resource manager of inpatient orders for targeted medications that frequently require prior authorization with the potential to delay discharge. A workflow process using the identification tool and flowsheet documentation was developed to prompt the initiation of a prior authorization, if necessary. Following hospital-wide implementation, descriptive data for a 2-month period was collected. Results: The tool detected 1353 medications for 1096 patient encounters over the 2-month period. The most frequent medications identified included apixaban (28.1%), enoxaparin (14.4%), sacubitril/valsartan (6.4%), and darbepoetin (6.4%). For the medications identified, there were 93 medications documented in the flowsheet data for 91 unique patient encounters. Of the 93 medications documented, 30% did not require prior authorization, 29% had prior authorization started, 10% were for patients discharged to a facility, 3% were for home medications, 3% were medications discontinued at discharge, 1% had prior authorization denied, and 24% had missing data. The most frequent medications documented in the flowsheet included apixaban (12%), enoxaparin (10%), and rifaximin (20%). Of the 28 prior authorizations processed, 2 led to a referral to the Medication Assistance Program. Conclusion: The implementation of an identification tool and documentation process can help improve PA workflow and discharge care coordination.

To plate, or not to plate? A systematic review of functional outcomes and complications of plate fixation in patellar fractures.

PURPOSE: Poor outcomes and high complication and reoperation rates have been reported with tension-band wiring (TBW) in the management of patellar fractures and particularly the comminuted ones. The purpose of this study was to investigate the functional outcomes and complication rates of patellar fractures managed with open reduction and internal fixation (ORIF) with a plate. METHODS: MEDLINE, EMCare, CINAHL, AMED and HMIC were searched, and the PRISMA guidelines were followed. Two independent reviewers extracted the data from the included studies and assessed them for the risk of bias. RESULTS: Plating of patellar fractures is associated with satisfactory range of movement (ROM) and postoperative function and low pain levels. We found a 10.44% complication rate and a low reoperation rate. Reoperations were mainly performed for metalwork removal. CONCLUSION: ORIF with plating of patellar fractures is a safe alternative in the management of patellar fractures and may be associated with a lower complication and reoperation rate compared to TBW. Future randomized prospective studies are needed to validated the results of the present systematic review.

The lectin-specific activity of Toxoplasma gondii microneme proteins 1 and 4 binds Toll-like receptor 2 and 4 N-glycans to regulate innate immune priming.

Infection of host cells by Toxoplasma gondii is an active process, which is regulated by secretion of microneme (MICs) and rhoptry proteins (ROPs and RONs) from specialized organelles in the apical pole of the parasite. MIC1, MIC4 and MIC6 assemble into an adhesin complex secreted on the parasite surface that functions to promote infection competency. MIC1 and MIC4 are known to bind terminal sialic acid residues and galactose residues, respectively and to induce IL-12 production from splenocytes. Here we show that rMIC1- and rMIC4-stimulated dendritic cells and macrophages produce proinflammatory cytokines, and they do so by engaging TLR2 and TLR4. This process depends on sugar recognition, since point mutations in the carbohydrate-recognition domains (CRD) of rMIC1 and rMIC4 inhibit innate immune cells activation. HEK cells transfected with TLR2 glycomutants were selectively unresponsive to MICs. Following in vitro infection, parasites lacking MIC1 or MIC4, as well as expressing MIC proteins with point mutations in their CRD, failed to induce wild-type (WT) levels of IL-12 secretion by innate immune cells. However, only MIC1 was shown to impact systemic levels of IL-12 and IFN-γ in vivo. Together, our data show that MIC1 and MIC4 interact physically with TLR2 and TLR4 N-glycans to trigger IL-12 responses, and MIC1 is playing a significant role in vivo by altering T. gondii infection competency and murine pathogenesis.

Safety and immunogenicity of 17DD attenuated yellow fever vaccine in howler monkeys (Alouatta spp.).

BACKGROUND: Alouatta spp. are highly susceptible to yellow fever (YF) infection and develop an often fatal disease. The threat posed by an outbreak started in 2016 leads us to investigate vaccination as a potential tool in preventing YF in non-human primates (NHP). METHODS: Susceptible howler monkeys were immunized with three different concentrations of the human Brazilian commercial YF17DD vaccine. Post-vaccination viremia/RNAemia, immunogenicity, and safety were characterized. RESULTS: The vaccine did not produce YF clinical manifestations in any of the NHPs. After immunization, all animals seroconverted demonstrating the ability of the YF vaccine to induce humoral response in Alouatta species. CONCLUSIONS: The present work has demonstrated the safe and immunogenic profile of the existing YF 17DD vaccine in howler monkeys. This knowledge may support further studies with other susceptible monkey species and provide a possible solution for controlling epizootics and preventing the devastation of endangered species.

On the Development of Selective Chelators for Cadmium: Synthesis, Structure and Chelating Properties of 3-((5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)amino)benzo[d]isothiazole 1,1-dioxide, a Novel Thiadiazolyl Saccharinate.

Aquatic contamination by heavy metals is a major concern for the serious negative consequences it has for plants, animals, and humans. Among the most toxic metals, Cd(II) stands out since selective and truly efficient methodologies for its removal are not known. We report a novel multidentate chelating agent comprising the heterocycles thiadiazole and benzisothiazole. 3-((5-(trifluoromethyl)-1,3,4-thiadiazol-2-yl)amino)benzo[d]isothiazole 1,1-dioxide (AL14) was synthesized from cheap saccharin and characterized by different techniques, including single crystal X-ray crystallography. Our studies revealed the efficiency and selectivity of AL14 for the chelation of dissolved Cd(II) (as compared to Cu(II) and Fe(II)). Different spectral changes were observed upon the addition of Cd(II) and Cu(II) during UV-Vis titrations, suggesting different complexation interactions with both metals.

DDX3 suppresses type I interferons and favors viral replication during Arenavirus infection.

Several arenaviruses cause hemorrhagic fever (HF) diseases that are associated with high morbidity and mortality in humans. Accordingly, HF arenaviruses have been listed as top-priority emerging diseases for which countermeasures are urgently needed. Because arenavirus nucleoprotein (NP) plays critical roles in both virus multiplication and immune-evasion, we used an unbiased proteomic approach to identify NP-interacting proteins in human cells. DDX3, a DEAD-box ATP-dependent-RNA-helicase, interacted with NP in both NP-transfected and virus-infected cells. Importantly, DDX3 deficiency compromised the propagation of both Old and New World arenaviruses, including the HF arenaviruses Lassa and Junin viruses. The DDX3 role in promoting arenavirus multiplication associated with both a previously un-recognized DDX3 inhibitory role in type I interferon production in arenavirus infected cells and a positive DDX3 effect on arenavirus RNA synthesis that was dependent on its ATPase and Helicase activities. Our results uncover novel mechanisms used by arenaviruses to exploit the host machinery and subvert immunity, singling out DDX3 as a potential host target for developing new therapies against highly pathogenic arenaviruses.

Effect of thermal stress on basic seminal characteristics of Gyr bulls.

The objective of this study was to evaluate the influence of thermal stress indices and minimum temperature on the semen quality of dairy Gyr bulls and to determine whether the minimum night temperature is able to balance the thermal stress that occurred during the day. A total of 922 semen samples were collected from 391 bulls aged between 15 and 49 months, which participated in progeny pretest trials, from December 2011 to April 2017. Semen characteristics (ejaculate volume, gross motility, sperm motility, sperm vigor, sperm concentration, and percentage of total defects) and scrotal circumference were analyzed. Meteorological data were recorded daily by the National Institute of Meteorology. The analyses were carried out using mixed models. The following fixed effects were included in the model: test, month of sample collection, scrotal circumference, the age of the bull at sampling as covariate (linear regression), and the climatic variables (equivalent temperature index [ETI], temperature and humidity index [THI], or minimum temperature). The effect of the bull nested within the test was included in the model as a random effect. Two periods were considered to evaluate the effect of the climatic variables on sperm quality: 7 days before collection (period 1) and the day of collection (period 2). There was an effect of ETI on sperm vigor and concentration in both periods and on volume and percentage of total defects only in the second period. The THI exerted a significant effect on volume in both periods and on sperm motility and concentration only in the second period. There was an effect of the minimum temperature on ejaculate volume, sperm motility, sperm concentration, and percentage of total defects only in the second period, which corresponds to the day of semen collection. Taken together, the results show that there was sufficient thermal stress to negatively affect semen quality. However, the minimum temperature during the night was a significant factor that balanced these negative effects of THI and ETI on semen traits of Gyr bulls.

Evaluation of the Katsuki-Sharpless Epoxidation Precatalysts by ESI-FTMS, CID, and IRMPD Spectroscopy.

The Katsuki-Sharpless epoxidation reaction is one of the most recognized chiral catalytic reactions, allowing for chiral epoxides to be used as starting materials for a series of synthetic pathways. The complete understanding of this reaction mechanism depends on the identification and description of species formed from Ti(IV) alkoxides and alkyl tartrates precatalysts. Despite previous reports on the nature of a bimetallic catalyst based on IR spectroscopy and NMR analysis, some debate remains. Therefore, we carried out mass spectrometry analysis by direct extraction of the ions from the reaction media by ESI(-)-FT-ICR and evaluated the observed ions by CID and IRMPD experiments. These techniques allowed us to detect carboxylates that correlate to the species in the reaction media and to confirm the actual presence of the titanium-tartrate complexes in solution. Our IRMPD results suggest the carboxylate dimer as an asymmetric species with two tartrates coordinated to a single Ti atom, while the other Ti center is coordinated by labile alkoxydes that could be easily exchanged by the organic peroxide and the substrate, allowing the epoxidation reaction to take place.

Genetic diversity and antimicrobial resistance profile of Enterococcus faecalis isolated from broilers with vertebral osteomyelitis in Southeast Brazil.

Vertebral osteomyelitis (VO) is a worldwide emerging disease that affects broilers. Recently, the isolation of Enterococcus faecalis in cases of the disease has been described. This study aimed at determining the genetic diversity and antimicrobial resistance profile of 12 E. faecalis strains isolated from broilers with VO. Strains were isolated from nine flocks from six farms in a high-density poultry production area in Southeast Brazil and were evaluated using multilocus sequence typing and phylogenetic analysis. Antimicrobial susceptibility tests and PCR were performed to detect antimicrobial resistance genes. E. faecalis isolates belonged to different sequence types (ST), six of which (ST49, ST100, ST116, ST202, ST249, and ST300) have been previously described. Strains ST708 and ST709 were newly identified in this study. Strain ST49 was most frequently isolated (50% of the flocks) from the analysed VO cases. No phylogenetic or phylogeographic relationship was found among the strains. The VO isolated E. faecalis strains showed highest resistance to aminoglycosides, mainly gentamicin (40%), but were highly susceptible to vancomycin (10%). Aminoglycoside resistance genes were detected in seven E. faecalis strains, and AAC6'-APH2″ genes were most frequently detected. The results showed that E. faecalis strains isolated from recently reported VO cases were highly diverse genetically. The diversity of genotypes in circulation in the analysed flocks, without apparent relationship among them, raises questions on aetiopathogenesis of the disease in broilers and evolutionary aspects of E. faecalis.

Method for the simultaneous determination of monoaromatic and polycyclic aromatic hydrocarbons in industrial effluents using dispersive liquid-liquid microextraction with gas chromatography-mass spectrometry.

We present a new method for simultaneous determination of 22 monoaromatic and polycyclic aromatic hydrocarbons in postoxidative effluents from the production of petroleum bitumen using dispersive liquid-liquid microextraction coupled to gas chromatography and mass spectrometry. The eight extraction parameters including the type and volume of extraction and disperser solvent, pH, salting out effect, extraction, and centrifugation time were optimized. The low detection limit ranging from 0.36 to 28 µg/L, limit of quantitation (1.1-84 µg/L), good reproducibility, and wide linear ranges, as well as the recoveries ranging from 71.74 to 114.67% revealed that the new method allows the determination of aromatic hydrocarbons at low concentration levels in industrial effluents having a very complex composition. The developed method was applied to the determination of content of mono- and polycyclic aromatic hydrocarbons in samples of raw postoxidative effluents in which 15 compounds were identified at concentrations ranging from 1.21 to 1017.0 µg/L as well as in effluents after chemical treatment.

Latex balloons: an alternative, low-cost model for vascular anastomosis training in medical education.

Simulators are increasingly being used in medical education, but accessibility is restricted by their elevated cost. A accessible and low-cost model was developed for teaching and learning vascular sutures and anastomoses at a Basic Surgical Techniques Laboratory. Latex balloons of varying colors, polypropylene 6.0 sutures, and other materials specifically for suturing (needle holder and forceps) were used. The balloons were fixed to screws inserted into wooden boards in order to facilitate repairs. E end-to-end, end-to-side, and side-to-side anastomoses and patching were performed. Anastomosis patency was tested by injecting water into one extremity of the balloon and observing the liquid exit via the opposite extremity. The advantages observed with this training model for anastomoses were malleability, resistance to passage of the suture, and the fact that it is inorganic. Latex balloons are an inexpensive option that are non­perishable and offer prolonged use for teaching and practice of arterial sutures and anastomoses.

Probing the geometry reorganization from solution to gas-phase in putrescine derivatives by IRMPD, 1H-NMR and theoretical calculations.

Single and double protonated (E)-1,4-diamine-2-butenes were evaluated as a model system to probe isomerization during the ESI processes employing infrared multiple-photon dissociation (IRMPD) spectroscopy and density function theory (DFT) calculations, including implicit and explicit solvation models. Our results show that the preferential protonation takes place at the amines for singly protonated species and that the double bond is not protonated even under double protonation, as expected from known pKa values. This behavior was shown to reflect the (E)-(Z) interconversion rate, as no interconversion was observed nor predicted by implicit solvation model based on density (SMD) calculations even by the olefin protonation pathway. Explicit solvent calculations show that the singly protonated (E) configuration observed in the gas phase is also the most stable configuration in solution due to molecular interactions with the solvent that are absent for the (Z) configuration. The explicit solvation calculation reverts the supposed gas-phase stability of the (Z) configuration in comparison to (E) from -9 kcal mol-1 (in relative Gibbs free energy) in the gas phase to +89 kcal mol-1 (in total potential energy) as depicted by explicit Monte Carlo (MC) simulations. Together with previous results for the saturated 1,4-diamines from Morton and coworkers that show the (Z) configuration related conformation to be the most stable geometry in the gas-phase due to intermolecular hydrogen bonding, our experiments clearly show that conformational reorganizations can take place during the ESI process. These results suggest that gas-phase experiments and vacuum calculations may not be valid as evidence for conformations in solution without prior testing to check for isomerization during the ESI process.

New procedure for the examination of the degradation of volatile organonitrogen compounds during the treatment of industrial effluents.

We present a new procedure for the determination of 32 volatile organonitrogen compounds in samples of industrial effluents with a complex matrix. The procedure, based on dispersive liquid-liquid microextraction followed by gas chromatography with nitrogen-phosphorus and mass spectrometric detection, was optimized and validated. Optimization of the extraction included the type of extraction and disperser solvent, disperser solvent volume, pH, salting out effect, extraction, and centrifugation time. The procedure based on nitrogen-phosphorus detection was found to be superior, having lower limits of detection (0.0067-2.29 µg/mL) and quantitation as well as a wider linear range. The developed procedure was applied to the determination of content of volatile organonitrogen compounds in samples of raw effluents from the production of bitumens in which 13 compounds were identified at concentrations ranging from 0.15 to 10.86 µg/mL and in samples of effluents treated by various chemical methods.