Influence of oblique proximal ulnar osteotomy on humeral intracondylar fissures in 35 spaniel breed dogs.

OBJECTIVE: To investigate the effects of oblique proximal ulnar osteotomy (PUO) on the healing of humeral intracondylar fissure (HIF) in spaniel breed dogs. STUDY DESIGN: Clinical retrospective study. SAMPLE POPULATION: A total of 51 elbows from 35 spaniel dogs. METHODS: An oblique PUO was performed in dogs diagnosed with HIF. The degree of healing of HIF was subjectively and objectively assessed on preoperative and long-term follow-up CT imaging. Objective assessment was performed by measuring the bone density in Hounsfield units (HU) of a rectangular region of interest (ROI) encompassing the entire hypoattenuated humeral fissure. Major and minor complications were recorded. RESULTS: A total of 24 partial and 27 complete HIFs were diagnosed. The follow-up CT scan was performed at a median 18.5 months (range 10-49 months). Subjective assessment confirmed partial or complete healing of the HIF in 41 elbows (80.3%). Objective assessment confirmed a difference in mean HU of the HIF's ROI between preoperative (HU 640) and last follow-up CT images (HU 835) (p = .001). Young dogs (<14 months) had the highest increase in HU of the HIF's ROI. Major complications occurred in five dogs (6 limbs) of which four were related to the lack of healing of the fissure (7.8%). CONCLUSION: Oblique PUO resulted in partial or complete healing of HIF and pain resolution in the majority of dogs. CLINICAL SIGNIFICANCE: This study introduces an innovative approach to achieve healing of the HIF in the dog, which may help reduce the high complication rate traditionally associated with the use of transcondylar screws.

Pesticides luminescent sensing by a Tb3+-doped Zn metal-organic framework with selectivity towards parathion.

Organophosphorus pesticides (OPPs) such as parathion have extensive uses in agriculture and household applications. Chronic exposure to these pesticides can cause severe health and environmental issues. Therefore, a current ecological concern is associated with accumulating these noxious OPPs in food and water sources. In this work, a new Tb3+-doped Zn-LMOF (Zn-LMOF= (3D) {[Zn3(1,4 benzenedicarboxylate)3(EtOH)2]·(EtOH)0.6}∞) was synthesized by a solvent-free reaction between the Zn-LMOF and the salt TbCl3·6H2O using a high-speed ball milling. The Tb@Zn-LMOF was thoroughly characterized by multiple spectroscopic tools, including Scanning Electron Microscopy with Energy-Dispersive X-ray Spectroscopy, and studied in-depth as a luminescent sensor for a series of pesticides (parathion, malathion, methalaxil, carbofuran, iprodione, captan and glyphosate) in aqueous methanol. The Tb@Zn-LMOF is a long-lived green-emitting compound with luminescence originated by an efficient antenna effect from the excited energy levels of Zn-LMOF toward the 5D state of Tb3+ ions, as it is displayed by its strong emission bands at 488, 545, 585, and 620 nm and a lifetime of 1.01 ms upon excitation at 290 nm. Additions of pesticides to a neutral methanolic dispersion of Tb@Zn-LMOF modified its green emission intensity with a pronounced selectivity toward parathion within the micromolar concentration range. The detection limit for parathion was calculated to be 3.04 ± 0.2 µM for Tb@Zn-LMOF. Based on 31P NMR and mass spectrometry studies, it is attributed to the release of lanthanide ions from Tb@Zn-LMOF with the simultaneous formation of a Tb3+-parathion complex.

Water-soluble fluorescent chemosensor for sorbitol based on a dicationic diboronic receptor. Crystal structure and spectroscopic studies.

Selective recognition of saccharides by phenylboronic dyes capable of functioning in aqueous conditions is a central topic of modern supramolecular chemistry that impacts analytical sciences and biological chemistry. Herein, a new dicationic diboronic acid structure 11 was synthesized, structurally described by single-crystal X-ray diffraction, and studied in-depth as fluorescent receptor for six saccharides in pure water at pH = 7.4. This dicationic receptor 11 has been designed particularly to respond to sorbitol and involves two convergent and strongly acidified phenyl boronic acids, with a pKa of 6.6, that operate as binding sites. The addition of sorbitol in the micromolar concentration range to receptor 11 induces strong fluorescence change, but in the presence of fructose, mannitol, glucose, lactose and sucrose, only moderate optical changes are observed. This change in emission is attributed to a static complexation photoinduced electron transfer mechanism as evidenced by lifetime experiments and different spectroscopic tools. The diboronic receptor has a high affinity/selectivity to sorbitol (K = 31 800 M-1) over other saccharides including common interfering species such as mannitol and fructose. The results based on 1H, 11B NMR spectroscopy, high-resolution mass spectrometry and density functional theory calculations, support that sorbitol is efficiently bound to 11 in a 1 : 1 mode involving a chelating diboronate-sorbitol complexation. Since the experimental B⋯B distance (5.3 Å) in 11 is very close to the calculated distance from the DFT-optimized complex with sorbitol, the efficient binding is attributed to strong acidification and preorganization of boronic acids. These results highlight the usefulness of a new diboronic acid receptor with a strong ability for fluorescent recognition of sorbitol in physiological conditions.

Dynamic allosteric networks drive adenosine A1 receptor activation and G-protein coupling.

G-protein coupled receptors (GPCRs) present specific activation pathways and signaling among receptor subtypes. Hence, an extensive knowledge of the structural dynamics of the receptor is critical for the development of therapeutics. Here, we target the adenosine A1 receptor (A1R), for which a negligible number of drugs have been approved. We combine molecular dynamics simulations, enhanced sampling techniques, network theory and pocket detection to decipher the activation pathway of A1R, decode the allosteric networks and identify transient pockets. The A1R activation pathway reveal hidden intermediate and pre-active states together with the inactive and fully-active states observed experimentally. The protein energy networks computed throughout these conformational states successfully unravel the extra and intracellular allosteric centers and the communication pathways that couples them. We observe that the allosteric networks are dynamic, being increased along activation and fine-tuned in presence of the trimeric G-proteins. Overlap of transient pockets and energy networks uncover how the allosteric coupling between pockets and distinct functional regions of the receptor is altered along activation. By an in-depth analysis of the bridge between activation pathway, energy networks and transient pockets, we provide a further understanding of A1R. This information can be useful to ease the design of allosteric modulators for A1R.

Lower body weight and increasing age are significant risk factors for complications following bi-oblique proximal ulnar osteotomy in dogs.

OBJECTIVE: To report and evaluate the risk factors for complications following bi-oblique proximal ulnar osteotomy (PUO) in dogs. ANIMALS: 82 client-owned dogs (93 limbs). METHODS: Clinical records of dogs treated with bi-oblique PUO over a 5-year period were reviewed. Postoperative radiographs were analyzed, and osteotomy location and angles were recorded; follow-up radiographs were reviewed to assess the degree of osteotomy healing. Complications were classified as minor, major, and catastrophic. Logistic regression analysis was performed to investigate possible associations between the predictor variables and occurrence of complications. RESULTS: 82 dogs (93 limbs) were included. Postoperative complications were documented in 39 limbs (13 major and 26 minor). The most common major complication was osteotomy nonunion (8 limbs), while the most common minor complication was delayed union (21 limbs). Statistical analysis revealed that lower body weight (P = .01) and older age (P = .04) were significantly associated with the development of postoperative complications. CLINICAL RELEVANCE: In this study, lighter and older dogs were more likely to develop complications following bi-oblique proximal ulnar osteotomy. We did not identify an association between osteotomy angle and location on complication rate. Careful patient selection is therefore required to reduce the incidence of postoperative complications after bi-oblique PUO.

Wearable Sports Technology Development in Costa Rica: Inertial Measurement Unit Integration for Real-Time External Load Monitoring / Desarrollo de tecnología deportiva portátil en Costa Rica: integración de una unidad de medición inercial para monitoreo de carga externa en tiempo real / Desenvolvimento de tecnologia desportiva portátil na Costa Rica: integração de uma unidade de medição por inércia para monitorização da carga externa em tempo real

Abstract: The increasing trend and projection for the use of wearable sensors technology in the general population have led to the development of different inertial measurement units (IMU) to monitor activity and motion variables during exercise or sports. This study aims to systematize the practical experience of developing an inertial measurement device to quantify external load in real time for runners. This article shows the installed capacity of Costa Rican higher education institutions to integrate existing sensors available in the international market in a single device to program it and use it for different sport science purposes. Two scientific laboratories joined resources and integrated an interdisciplinary team composed of exercise physiologists, sports scientists, physiotherapists, and electromechanical engineers who globally faced the challenge of creating SafeRun IMU. In this case, an IMU was constructed with great success, capable of quantifying external load variables controlled from a mobile device in real-time. The results showed an acceptable agreement and reliability of the data extracted from the IMU. The information collected from this IMU could be used to control and quantify external load to develop or redirect exercise programming and prescription and prevent overuse injuries or physical overload conditions. This device differs from other existing ones due to its small size and portability, which benefits its commercialization and massification.

Resumen: La creciente tendencia y proyección para el uso de tecnología de sensores portátiles en la población en general ha llevado al desenvolvimiento de diferentes dispositivos de medición inercial, para monitorear las variables de actividad y movimiento durante el ejercicio o los deportes. El objetivo del presente estudio es sistematizar la experiencia práctica de desarrollar un dispositivo de medición inercial, con el fin de cuantificar la carga externa en tiempo real para los corredores. Este artículo demostró la capacidad instalada de las instituciones de educación superior de Costa Rica para integrar los sensores existentes disponibles en el mercado internacional en un solo dispositivo, con el propósito de programarlo y usarlo en diferentes fines de ciencias del deporte. Dos laboratorios científicos unieron esfuerzos e integraron un equipo interdisciplinario compuesto por fisiólogos del ejercicio, científicos del deporte, fisioterapeutas e ingenieros electromecánicos, quienes, de manera global, afrontaron el reto de la creación de la IMU. En este caso, se desarrolló una IMU con gran éxito, capaz de cuantificar las variables de carga externa controladas desde un dispositivo móvil en tiempo real. Los resultados mostraron un nivel de concordancia y fiabilidad aceptable de los datos extraídos de la IMU. La información obtenida de esta IMU podría usarse para controlar y cuantificar la carga externa, con miras a desplegar o redirigir la programación y prescripción de ejercicios, así como con el afán de prevenir lesiones por sobreuso o condiciones de sobrecarga física.

Resumo: A crescente tendência e projeção para o uso da tecnologia de sensores portáteis na população em geral levou ao desenvolvimento de diferentes dispositivos de medição inercial para monitorar as variáveis de atividade e de movimento durante o exercício ou esportes. O objetivo do presente estudo é sistematizar a experiência prática de desenvolvimento de um dispositivo de medição inercial, a fim de quantificar a carga externa em tempo real para os corredores. Este artigo demonstrou a capacidade instalada das instituições de ensino superior da Costa Rica para integrar os sensores existentes disponíveis no mercado internacional em um único dispositivo, com a finalidade de programá-lo e usá-lo para diferentes fins de ciência do esporte. Dois laboratórios científicos uniram forças e integraram uma equipe interdisciplinar composta por fisiologistas do exercício, cientistas do esporte, fisioterapeutas e engenheiros eletromecânicos, que, de forma global, enfrentaram o desafio de criar a IMU. Neste caso, foi desenvolvida uma IMU de grande sucesso, capaz de quantificar variáveis de carga externas controladas a partir de um dispositivo móvel em tempo real. Os resultados mostraram um nível aceitável de concordância e confiabilidade dos dados extraídos da IMU. As informações obtidas desta IMU poderiam ser utilizadas para monitorar e quantificar a carga externa, com o objetivo de implantar ou redirecionar a programação e prescrição de exercícios, bem como com o objetivo de evitar lesões por uso excessivo ou condições de sobrecarga física.

The occurrence of Sarcocystis spp. in the myocardium of alpacas (Vicugna pacos) with associated risk factors in the Peruvian Andes.

Sarcocystis masoni n. sp. (known as "S. lamacanis") infects alpacas affecting their productivity and can cause a food poisoning syndrome in humans by consuming contaminated, undercooked cardiac muscle. There are few studies estimating the prevalence of this parasite in alpacas, although this information is crucial for the control and prevention of sarcocystosis. This study aimed to determine the frequency and density of Sarcocystis masoni n. sp. in the heart of alpacas in Huancavelica, a province of the Andean region of Peru. Heart samples were taken for histopathology from 104 alpacas slaughtered at the municipal slaughterhouse of Huancavelica, the official abattoir in the Huancavelica district. No macroscopic sarcocysts were observed. All alpacas (100%) had microscopic sarcocysts of Sarcocystis masoni n. sp., with no inflammatory reactions. The alpacas showed an average sarcocyst density of 60.8 ± 23.3/mm2. Sarcocysts density was significantly higher (p < 0.05) as the age of the animals increased. In addition, sarcocysts density was significantly higher (p < 0.05) in male animals aged 4 and 5 years compared to females of the same age. These results confirmed that heart sarcocystosis is highly endemic in Peruvian alpacas. Therefore, it is recommended that alpaca hearts be well-cooked at the time of consumption. The present study showed current data and contributes to the knowledge of this parasitosis. Studies of this nature are necessary because they are the basis for developing animal health programs.

Chemical Characterization, Antioxidant Capacity and Anti-Oxidative Stress Potential of South American Fabaceae Desmodium tortuosum.

It has been proposed that oxidative stress is a pathogenic mechanism to induce cytotoxicity and to cause cardiovascular and neuronal diseases. At present, natural compounds such as plant extracts have been used to reduce the cytotoxic effects produced by agents that induce oxidative stress. Our study aimed to evaluate the antioxidant and cytoprotective capacity of Desmodium tortuosum (D. tortuosum) extract in the co- and pre-treatment in EA.hy926 and SH-SY5Y cell lines subjected to oxidative stress induced by tert-butylhydroperoxide (t-BOOH). Cell viability, reactive oxygen species (ROS), nitric oxide (NO), caspase 3/7 activity, reduced glutathione (GSH), glutathione peroxidase (GPx), glutathione reductase (GR), and molecular expression of oxidative stress biomarkers (SOD2, NRF2 and NFκB1) and cell death (APAF1, BAX, Caspase3) were all evaluated. It was observed that the D. tortuosum extract, in a dose-dependent manner, was able to reduce the oxidative and cytotoxicity effects induced by t-BOOH, even normalized to a dose of 200 µg/mL, which would be due to the high content of phenolic compounds mainly phenolic acids, flavonoids, carotenoids and other antioxidant compounds. Finally, these results are indicators that the extract of D. tortuosum could be a natural alternative against the cytotoxic exposure to stressful and cytotoxic chemical agents.

Locking Compression Plate Fixation of Feline Acetabular Fractures: Application, Complications and Perioperative Outcome.

OBJECTIVES: The aim of this study was to report the use, complications and perioperative outcome of locking compression plates (LCP) for acetabular fracture fixation in feline patients. STUDY DESIGN: Medical records were reviewed for cats presented with acetabular fractures between 2011 and 2019. Only cases that involved open reduction and internal fixation with at least one LCP and had a minimum of 5-week postoperative follow-up were included. Long-term follow-up was based on the Feline Musculoskeletal Pain Index. RESULTS: Fifteen cats with 15 acetabular fractures met the inclusion criteria. Locking compression plates were used as a sole method of fixation in 11/15 cases, while with other ancillary implants in 4/15 cases. There were two minor complications related to the LCP consisting of a single screw backing out. Two major perioperative complications not related to the locking implants were described and required further surgical treatment. At the last clinical follow-up (median 46 days: 38-88 days), function was described as full for 7/15 cats, and acceptable for 8/15 cats. The Feline Musculoskeletal Pain Index questionnaire was completed by 9/15 owners at a median of 2,332 days; all owners reported their cats had minimal impairment. CLINICAL SIGNIFICANCE: Based on our results, LCP is a suitable implant for acetabular fracture fixation in cats, associated with a satisfactory outcome. Complications were similar to previously reported with other methods.

Functional outcomes of dogs undergoing shoulder arthrodesis with 2 locking compression plates.

OBJECTIVE: To report the outcomes of dogs undergoing shoulder arthrodesis with the application of 2 locking compression plates (LCPs). STUDY DESIGN: Retrospective case series. ANIMALS: Twelve client owned dogs undergoing shoulder arthrodesis using 2 LCPs as the principal method of fixation. METHODS: Medical records were searched for dogs who had undergone shoulder arthrodesis at a hospital between 2014-2021. Inclusion criteria required at least 6 months postoperative follow up and the use of 2 LCPs as the principal method of arthrodesis. Outcomes recorded include clinical assessment, Liverpool Osteoarthritis in Dogs (LOAD) scoring, force plate analysis, and a client visual-analogue scale with at least 6 months follow up. RESULTS: All shoulders were immobilized with 2 LCPs and no postoperative external coaptation. The duration of follow up was 855.9 days (174-2135). Minor complications were recorded in 2/12 dogs, neither requiring surgical revision. Preoperative to postoperative mean lameness scores reduced from 2.9 to 0.8 (0-5), with 11/12 dogs making a full functional recovery, with a mean long-term LOAD score of 12.9 (4-23). Gait analysis was available in 5/12 dogs, placing a mean of 31.9% of their bodyweight on the operated limb, with a mean symmetry index of 8.7%. Owners scores (n = 11/12) were consistent with a high level of satisfaction (9.6/10). CONCLUSION: Shoulder arthrodesis with 2 LCPs was associated with low morbidity and good outcomes in our clinical setting. CLINICAL SIGNIFICANCE: Canine shoulder arthrodesis performed with 2 LCPs should be considered as a suitable alternative in dogs with severe shoulder joint pathology.

The Effect of Femur Positioning on Measurement of Tibial Plateau Angle: An In Vitro Study.

Five canine cadaveric pelvic limbs with intact cranial cruciate ligaments were used to quantify the effect of variation in limb positioning on the radiographic measurement of the tibial plateau angle (TPA) with reference to the degree of femoral condyle superimposition. Intra-osseous pin placement and a custom jig design allowed the controlled three-dimensional manipulation of limbs. Medio-lateral digital radiographic projections were taken with perfect femoral hemicondylar superimposition to establish a "reference" TPA (difference in position = 0 mm), and subsequently in varying degrees of supination/pronation and abduction/adduction. The lack of femoral hemicondylar superimposition for each radiograph was quantified using a tangential line technique with reference to the long tibial axis. A total of 176 radiographs were each assessed by three observers. "True" TPA was measured and it ranged within 17-25° across all limbs assessed. Variation in femoral condylar positioning ranged from -13 mm to +13 mm proximo-distally, and -11 mm to +11 mm cranio-caudally. Moreover, 3 mm non-superimposition of the femoral condyles produced 90.6% of measurements with 1° difference between measured and "true" TPA, and a sensitivity of 97.9% for a 2° difference. Further reduction in femoral condylar superimposition to 4 mm reduced the frequency of 1° difference between measured and "true" TPA to 84.9%, and to 94.8% for a 2° difference. In conclusion, measurement of TPA in large breed dogs from radiographs with greater than 3 mm variation in femoral condylar superimposition should be interpreted with caution.

Identification, Quantification, and Characterization of the Phenolic Fraction of Brunfelsia grandiflora: In Vitro Antioxidant Capacity.

Brunfelsia grandiflora is an ancient plant widely used for its promising medicinal properties, although little explored scientifically. Despite being a rich source of phenolic compounds responsible in part for the proven anti-inflammatory activity, its characterization has not been carried out to date. The present work deals with the exhaustive identification and quantification of its phenolic fraction, along with its antioxidant activity. Decoction resulting from the bark as fine powder was filtered and lyophilized, and polyphenols were extracted from the resulting product by aqueous-organic solvents. Seventy-nine polyphenols were identified using LC-MSn. Hydroxycinnamates was the most abundant group of compounds (up to 66.8%), followed by hydroxycoumarins (15.5%), lignans (6.1%), flavonols (5.7%), phenolic simples (3.1), gallates (2.3%), flavanols (0.3%), and flavanones (0.2%). About 64% of the characterized phenols were in their glycosylated forms. The quantification of these phytochemicals by LC-QToF showed that this medicinal plant contained 2014.71 mg of phenolic compounds in 100 g dry matter, which evidences a great antioxidant potency determined by ABTS and DPPH assays. Therefore, Brunfelsia grandiflora represents an important source of polyphenols which supports its therapeutic properties scientifically proven.

Time Evolution of the Millisecond Allosteric Activation of Imidazole Glycerol Phosphate Synthase.

Deciphering the molecular mechanisms of enzymatic allosteric regulation requires the structural characterization of functional states and also their time evolution toward the formation of the allosterically activated ternary complex. The transient nature and usually slow millisecond time scale interconversion between these functional states hamper their experimental and computational characterization. Here, we combine extensive molecular dynamics simulations, enhanced sampling techniques, and dynamical networks to describe the allosteric activation of imidazole glycerol phosphate synthase (IGPS) from the substrate-free form to the active ternary complex. IGPS is a heterodimeric bienzyme complex whose HisH subunit is responsible for hydrolyzing glutamine and delivering ammonia for the cyclase activity in HisF. Despite significant advances in understanding the underlying allosteric mechanism, essential molecular details of the long-range millisecond allosteric activation of IGPS remain hidden. Without using a priori information of the active state, our simulations uncover how IGPS, with the allosteric effector bound in HisF, spontaneously captures glutamine in a catalytically inactive HisH conformation, subsequently attains a closed HisF:HisH interface, and finally forms the oxyanion hole in HisH for efficient glutamine hydrolysis. We show that the combined effector and substrate binding dramatically decreases the conformational barrier associated with oxyanion hole formation, in line with the experimentally observed 4500-fold activity increase in glutamine hydrolysis. The allosteric activation is controlled by correlated time-evolving dynamic networks connecting the effector and substrate binding sites. This computational strategy tailored to describe millisecond events can be used to rationalize the effect of mutations on the allosteric regulation and guide IGPS engineering efforts.

Marginal excision of cutaneous mast cell tumors in dogs was not associated with a higher rate of complications or prolonged wound healing than marginal excision of soft tissue sarcomas.

OBJECTIVE: To compare wound healing following planned marginal excision of cutaneous mast cell tumors (MCTs) with that of soft tissue sarcomas (STSs) and to identify risk factors for wound healing complications and delay in healing. ANIMALS: 126 dogs that underwent intentional marginal excision of cutaneous MCTs (n = 77) or subcutaneous STSs (49). PROCEDURES: Medical records of included dogs were reviewed and signalment, tumor size, tumor location, skin closure type, time to healing, reported complications, histopathological grade, and surgical margins were recorded. These variables and outcomes (complication rate and time to complete healing) were compared between dogs in the MCT and STS groups. Potential risk factors for complications and delayed healing were analyzed. RESULTS: No significant difference between the groups was found in any of the variables. Wound healing complication rates were 29% (22/77) for the MCT group and 31% (15/49) for the STS group. The mean ± SD time to complete healing was 16.5 ± 7.5 days for the MCT group and 17.7 ± 9.3 days for the STS group. These outcomes did not differ significantly between groups. For both groups, the use of subdermal plexus flap reconstruction was associated with the development of complications and increased time to complete healing. CLINICAL RELEVANCE: Marginal excision of cutaneous MCTs was not associated with a higher rate of complication or prolonged wound healing, compared with marginal excision of STSs. The use of flap reconstruction in skin closure may delay healing and planned adjuvant therapy. Owners should be counseled regarding these risks and where appropriate and feasible, surgery without reconstruction should be considered.

Big data and artificial intelligence (AI) methodologies for computer-aided drug design (CADD).

There have been numerous advances in the development of computational and statistical methods and applications of big data and artificial intelligence (AI) techniques for computer-aided drug design (CADD). Drug design is a costly and laborious process considering the biological complexity of diseases. To effectively and efficiently design and develop a new drug, CADD can be used to apply cutting-edge techniques to various limitations in the drug design field. Data pre-processing approaches, which clean the raw data for consistent and reproducible applications of big data and AI methods are introduced. We include the current status of the applicability of big data and AI methods to drug design areas such as the identification of binding sites in target proteins, structure-based virtual screening (SBVS), and absorption, distribution, metabolism, excretion and toxicity (ADMET) property prediction. Data pre-processing and applications of big data and AI methods enable the accurate and comprehensive analysis of massive biomedical data and the development of predictive models in the field of drug design. Understanding and analyzing biological, chemical, or pharmaceutical architectures of biomedical entities related to drug design will provide beneficial information in the biomedical big data era.

Complications after proximal abducting ulnar osteotomy and prognostic factors in 66 dogs.

OBJECTIVE: To report complications and prognostic factors in dogs undergoing proximal abducting ulnar osteotomy (PAUL). To evaluate the ability to predict complications on the basis of post-operative radiographic examination. STUDY DESIGN: Retrospective cohort study. ANIMALS: Sixty-six dogs. METHODS: Medical records of dogs treated with PAUL between 2014 and 2019 were reviewed for demographics, intraoperative findings, and post-operative complications. Post-operative radiographs were reviewed by two masked expert orthopedic surgeons, who were asked to predict the likelihood of major mechanical complications. The prognostic value of variables was tested with univariate and multivariable logistic regression. Inter-investigator agreement to predict complications was evaluated with two-by-two tables and kappa coefficient. RESULTS: Seventy-four PAULs in 66 dogs were included. Duration of follow-up ranged from 12 to 75 months (median: 53 months). Post-operative complications were documented in 19/74 limbs (16 dogs), including major complications in 13 limbs. These complications consisted mainly of non-union (six limbs), implant failure (two limbs), and infection (two limbs) requiring revision surgery in nine limbs. Body weight was the only variable associated with an increased risk of post-operative complications (p = .04). Agreement between expert predictions was low (respectively k = -0.08 and k = 0.11). CONCLUSION: Major complications were reported in one fourth of limbs treated with PAUL and were more likely as body weight increased. Suboptimal plate and screw placement or osteotomy reduction on post-operative radiographs were poorly predictive of complications. CLINICAL SIGNIFICANCE: Complications are fairly common after PAUL, particularly in heavier dogs, and post-operative radiographic examination seems unreliable to predict those.

In Silico Identification and Experimental Validation of Distal Activity-Enhancing Mutations in Tryptophan Synthase.

Allostery is a central mechanism for the regulation of multi-enzyme complexes. The mechanistic basis that drives allosteric regulation is poorly understood but harbors key information for enzyme engineering. In the present study, we focus on the tryptophan synthase complex that is composed of TrpA and TrpB subunits, which allosterically activate each other. Specifically, we develop a rational approach for identifying key amino acid residues of TrpB distal from the active site. Those residues are predicted to be crucial for shifting the inefficient conformational ensemble of the isolated TrpB to a productive ensemble through intra-subunit allosteric effects. The experimental validation of the conformationally driven TrpB design demonstrates its superior stand-alone activity in the absence of TrpA, comparable to those enhancements obtained after multiple rounds of experimental laboratory evolution. Our work evidences that the current challenge of distal active site prediction for enhanced function in computational enzyme design has become within reach.

Clinical Cohort Study in Canine Patients, to Determine the Average Platelet and White Blood Cell Number and Its Correlation with Patient's Age, Weight, Breed and Gender: 92 Cases (2019-2020).

Due to its easy preparation and that it is well tolerated, the use of autologous platelet-rich plasma (PRP) has become increasingly popular in regenerative medicine. However, there are still no clear guidelines on how it should be classified or whether the individual canine patient's clinical status can influence its quality. OBJECTIVE: This study aims to show if the weight, age, sex, neutered status or breed of canine patients have any correlation with the composition of PRP. DESIGN: A blinded count of the platelets and white blood cells (WBC) was performed from 111 samples from 92 client owned dogs undergoing treatment for degenerative joint disease (DJD). The results were analysed using Pearson correlation test, ANOVA test or Student T-test. RESULTS: There is a positive correlation between the number of platelets and WBC in canine patients of different breeds, but there was no significant difference on the platelet number and WBC number among the different breeds. The weight of the patient is also directly correlated to the platelet number (p = 0.003) but not WBC number. WBC number was negatively correlated to the weight of the patient. The sex and age of the patient did not affect platelets and WBC number, although WBC number is increased in non-neutered male population (p = 0.003). However, it would be interesting to investigate whether the growth factors released from the platelet granules are affected by patient variables in a canine population. CONCLUSIONS: Our results show that it is possible to obtain good quality autologous PRP, irrespective of age, sex, neutered status or weight of the patient, for PRP regenerative therapy.

Optimization of the ultrasoud-assisted extraction of saponins from quinoa (Chenopodium quinoa Wild) using response surface methodology.

BACKGROUND: Quinoa grain has a bitter tasting layer in the pericarp called saponin, a triterpenoid glycoside with industrial potential. Traditionally, quinoa saponins are extracted with a large amount of water, which is why ultrasound technology constitutes an emerging technological alternative which is considered efficient and profitable compared to traditional extraction methods. The objective of this research was to determine the amplitude, time, and concentration of ethanol that guarantee a higher content of saponin through extraction assisted by ultrasound. METHODS: To find the optimal extraction conditions, the response surface methodology was used using the Box Behnken design with 5 central points, taking as a response the content of saponins (expressed in oleanolic acid as it is the most abundant sapogenin). RESULTS: According to the results obtained, the R2 values were in agreement with the adjusted R2, showing that the data fit the model well. The results showed that ethanol concentration has a significant effect (p < 0.05) on the saponin content in the extract. Optimization showed that the optimal extraction conditions were 70% ethanol, 59% amplitude and an exposure time of 12 min. These values were obtained experimentally to compare theoretical values and found residual error percentages less than 3%. The emulsifying activity was evaluated, reporting a value of 52,495 units of emulsion activity per milliliter (UAE/mL), and the foaming stability indicated that 87.54% of the initial foam was maintained after 5 min, indicating high stability. CONCLUSIONS: The parameters of ethanol concentration, amplitude and time were optimized in the extraction of saponins, assisted by ultrasound. Furthermore, the extract obtained had good foaming and emulsifying characteristics, suggesting its suitability for use in industry.

Comparison of Fat Harvested from Flank and Falciform Regions for Stem Cell Therapy in Dogs.

BACKGROUND: Adipose tissue has recently gained attention as a source of mesenchymal stem cells (AdMSCs) for applications in treating degenerative joint disease in veterinary patients. This study aimed to quantify the stromal vascular fractions (SVFs) and colony forming units (CFU) of AdMSCs from the falciform and flank regions and compare dogs of different ages and weights. METHODS: Fat tissue was harvested from the flank (21 dogs) and falciform regions (17 dogs). The fat tissue was enzymatically digested and the number of nucleated cells in the SVF was counted. The SVF was cultured in vitro and the cell growth was assessed by counting the CFU per gram of fat and the aspect ratio of the cells. CONCLUSIONS: There was no significant difference in the number of nucleated cells in the SVF from the two sites. The CFU/g of fat from falciform was 378.9 ± 293 g and from flank was 486.8 ± 517 g, and this was also insignificant. Neither age nor weight of the patient had an impact on the SVF or CFU/g. No surgical complications were reported from either of the sites. Harvesting fat for stem cell therapy for intra-articular therapy of degenerative joint disease can be an easy and fast process when obtaining the fat either from the flank or the falciform region, and it is not age or weight dependent. The harvest site for clinical canine patients can be left to the surgeon's discretion and comfort.