Exploiting Leishmania-Primed Dendritic Cells as Potential Immunomodulators of Canine Immune Response.

Dendritic cells (DCs) capture pathogens and process antigens, playing a crucial role in activating naïve T cells, bridging the gap between innate and acquired immunity. However, little is known about DC activation when facing Leishmania parasites. Thus, this study investigates in vitro activity of canine peripheral blood-derived DCs (moDCs) exposed to L. infantum and L. amazonensis parasites and their extracellular vesicles (EVs). L. infantum increased toll-like receptor 4 gene expression in synergy with nuclear factor κB activation and the generation of pro-inflammatory cytokines. This parasite also induced the expression of class II molecules of major histocompatibility complex (MHC) and upregulated co-stimulatory molecule CD86, which, together with the release of chemokine CXCL16, can attract and help in T lymphocyte activation. In contrast, L. amazonensis induced moDCs to generate a mix of pro- and anti-inflammatory cytokines, indicating that this parasite can establish a different immune relationship with DCs. EVs promoted moDCs to express class I MHC associated with the upregulation of co-stimulatory molecules and the release of CXCL16, suggesting that EVs can modulate moDCs to attract cytotoxic CD8+ T cells. Thus, these parasites and their EVs can shape DC activation. A detailed understanding of DC activation may open new avenues for the development of advanced leishmaniasis control strategies.

Comparing follow-up of patients with vitamin K antagonists in a health center: pre- and post-COVID-19 pandemic / Comparação do seguimento de doentes hipocoagulados com antagonistas da vitamina K numa unidade de saúde: pré e pós pandemia COVID-19

Introduction and objectives: During the COVID-19 pandemic, the follow-up of patients treated with vitamin K antagonists (VKAs) may have been affected. This study aims to compare how these patients were monitored pre- and post-COVID-19 pandemic and understand the impact of non-face-to-face appointments on their follow-up. Methods: We conducted a retrospective cohort study in a Portuguese Health Center. The study included patients treated with VKAs and followed at the Health Center for international normalized ratio (INR) monitoring between March 2019 and March 2021. Data collected: sex, age, type of VKA; INR; date of INR assessment, type of appointment (face-to-face or phone/e-mail). Rosendaal's method was used to calculate pre-COVID-19 and post-COVID-19 time in therapeutic range (TTR). Good TTR control was defined if values ≥ 70%. Results: 44 patients were included. The mean TTR in the pre-COVID-19 period was 64.55% (95% CI: 58.10 - 71.00%). The post-COVID-19 mean was slightly higher (+ 2.26%), 66.81% (95% CI: 59.66 - 73.97%), but the difference was not statistically significant (p = 0.576). The use of non-face-to-face appointments did not contribute to worsening post-pandemic TTR, show-ing no lower follow-up than during pre-pandemic period in which all contacts were face-to-face [CI (95%) -0.397 - 0.196 for a reference range -0.489 - 0.693]. Conclusions: The TTR value in both periods was similar and lower than the value defined for effective hypocoagulation. The use of non-face-to-face consultation in the post-COVID-19 period does not seem to have influenced the quality of hypocoagulation (AU).

Introdução e objetivos: Durante a pandemia COVID-19 o acompanhamento de doentes medicados com antagonistas da vitamina K (AVKs) pode ter sido afetado. Este estudo pretende comparar a forma como estes doentes foram monitorizados antes e depois da pandemia COVID-19 e compreender o impacto da consulta não presencial no seu seguimento. Métodos: Estudo de coorte retrospetivo num Centro de Saúde em Portugal. O estudo incluiu doentes tratados com AVKs e seguidos no Centro de Saúde para monitorização do International Normalized Ratio(INR) entre março de 2019 e março de 2021. Dados recolhidos: sexo, idade, tipo de AVK; INR; data da avaliação do INR, tipo de consulta (presencial ou por telefone/e-mail). Foi utilizado o método de interpolação linear de Rosendaal para calcular o tempo em intervalo terapêutico (TTR) pré- e pós-COVID-19. Foi definido um bom controle se valores de TTR ≥ 70%. Resultados: Foram incluídos 44 doentes. A média de TTR no período pré-COVID-19 foi de 64,55% (95% IC: 58,10 - 71,00%). A média pós-COVID-19 foi ligeiramente superior (+ 2,26%), 66,81% (95% IC: 59,66 - 73,97%), mas a diferença não foi estatisticamente significativa (p = 0,576). A utilização da consulta não presencial não contribuiu para o agravamento do TTR no período pós-pandemia, não mostrando um seguimento inferior ao do período pré-pandemia em que todos os contatos foram presenciais [IC (95%) -0,397 - 0,196 para um intervalo de referência -0,489 - 0,693]. Conclusões: O valor de TTR em ambos os períodos foi semelhante e inferior ao valor definido para hipocoagulação eficaz. A utilização da consulta não presencial no período pós-COVID-19 não parece ter influenciado a qualidade da hipocoagulação (AU).

Training Effects on the Stress Predictors for Young Lusitano Horses Used in Dressage.

The purpose of this study was to investigate stressful responses during a 6-week training protocol in young Lusitano horses used for dressage. The hypothesis was that the proposed training protocol would improve fitness and ensure the welfare of the animals by reducing stress predictors. Nine 4-year-old horses were evaluated before (M1) and six weeks after (M2) beginning a training protocol. The training program was performed six times per week and included 40−80 min of individually intensity-adjusted preparatory exercises for dressage. For both moments, the horses were examined before (T0) and after (T1) dressage simulation tests (DST), and at 30 (T2) and 240 min (T3) during the recovery period. Blood samples were taken to determine the horses' cortisol levels, total WBC, and neutrophil and lymphocyte counts. All variables were analyzed by one-way ANOVA and Tukey tests, with p ≤ 0.05. After training, there was a significant reduction in cortisol (p = 0.0133), HR (p = 0.0283), total WBC (p < 0.0001), and neutrophil (p < 0.0001) and lymphocyte (p = 0.0341) counts. Other findings included an increase in HRV parameters related to a cardiac vagal modulation. In conclusion, the chosen training protocol led to better fitness as the horses worked more intensively with lower cardiovascular requirements, and they showed blunted cortisol responses at M2. Such data can be used to evaluate performance, but also to predict the welfare of athletic horses.

A survey on the orthopedic and functional assessment in a Portuguese population of police working dogs.

BACKGROUND: Working dogs are at an increased risk of developing an orthopedic disease compared to companion dogs. This study aimed to evaluate functional and orthopedic index fitness in a Portuguese population of police working dogs. In an observational, prospective study, information on 165 dogs was collected. The age, sex, breed, specific work, and history of previous diagnosis of orthopedic disease were recorded for each patient. A copy of the Canine Orthopedic Index (COI), Hudson Visual Analogue Scale (HVAS), and Functional Assessment (FA) was collected for all dogs. COI, HVAS, and FA scores between breeds, work, age, sex, and history of a previous diagnosis of orthopedic disease were compared. Multiple regression was run to predict COI, HVAS, and FA scores from breeds, work, age, sex, and history of orthopedic disease. Correlations between items were determined with Pearson's correlation. A p < 0.05 was set. RESULTS: The sample was composed of 92 males and 73 females, with a mean age of 5.2 ± 3.2 years. Four main dog breeds were represented, 60 Belgian Malinois Shepherd Dogs, 52 German Shepherd Dogs, 29 Labrador Retrievers, and 14 Dutch Shepherd Dog. A prevalence of diarrhea of 10.6% was determined, with 4% of dogs having liquid diarrhea. German Shepherd Dogs had significantly higher FA scores (p = 0.03). Dogs with a history of previous veterinary assistance due to orthopedic issues had significantly lower HVAS scores and higher scores with all remaining questionnaires (p < 0.01 for all). No differences were found between sexes or specific work. Age and a history of orthopedic disease contributed to the prediction of all scores. FA scores had a good correlation with COI and HVAS. CONCLUSION: This population of police working dogs has a good to excellent level of physical fitness. There was a relationship between increasing age, history of orthopedic disease, and worse scores with all questionnaires. All considered questionnaires could differentiate between animals with a previous history of orthopedic disease and sound dogs.

Effect of a single intra-articular high molecular weight hyaluronan in a naturally occurring canine osteoarthritis model: a randomized controlled trial.

BACKGROUND: Osteoarthritis (OA) is a complex joint disease and chronic pain source, affecting a patient's quality of life and posing a financial burden. As the dog is considered a nearly ideal species for translation research of human OA and the most used model for research, exploring spontaneous dog OA under the One Health/One Medicine concept can improve both humans and dogs' health and well-being. METHODS: In a clinical treatment experiment, forty (N=40) joints were selected and randomly assigned to a control group (CG), which received 0.9% NaCl or a treatment (HG), which received Hylan G-F 20. Evaluations were performed on treatment day (T0), 8, 15, 30, 90, and 180 days post-treatment. They consisted of four different Clinical Metrology Instruments (CMI), evaluation of weight distribution, joint range of motion, thigh girth, radiographic and digital thermography imaging, synovial fluid interleukin-1 (IL-1), and C-reactive protein concentrations. Results were compared with repeated measures ANOVA, with a Huynh-Feldt correction, Paired samples T-test, or Wilcoxon signed-ranks test, with p<0.05. RESULTS: Patients had a mean age of 6.5±2.4 years and a bodyweight of 26.6±5.2kg, and joints graded as mild (n=28, 70%), moderate (n=6, 15%), and severe OA (n=6, 15%). No differences were found between groups at T0. Symmetry index and deviation showed significant improvements in HG from 30 days (p<0.01) up to 180 days (p=0.01). Several CMI scores, particularly pain scores, improved from 90 to 180 days. Radiographic signs progressed in both groups. In both groups, increasing body weight and age corresponded to worse clinical presentation. IA hyaluronan administration produced increased lameness in six cases, which resolved spontaneously. CONCLUSIONS: This study characterizes the response to treatment with Hylan G-F 20, which can produce significant functional and pain level improvements in patients with OA, even those with factors related to worse response to treatment.

Management of Osteoarthritis Using 1 Intra-articular Platelet Concentrate Administration in a Canine Osteoarthritis Model.

BACKGROUND: Osteoarthritis (OA) represents a significant burden to societies, as it affects quality of life and performance and implies a large cost in terms of health care. Autologous platelets are a regenerative treatment modality for OA that are thought to be a potential stimulation of the natural healing cascade. PURPOSE: To describe the effect of the platelet concentrate V-PET in the management of OA in a naturally occurring canine model, using several outcome assessment modalities. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 40 joints of active working police dogs with hip OA were randomly assigned to a control group (CG) and a platelet concentrate group (PCG; treatment) and evaluated. At treatment day (T0) and 8, 15, 30, 90, and 180 days after treatment, weight distribution, joint range of motion at flexion and extension, thigh girth, digital thermography, radiographic signs, 4 clinical metrology instruments, and synovial fluid interleukin 1 and C-reactive protein levels were recorded. Results were compared using repeated-measures analysis of variance with a Huynh-Feldt correction, paired-samples t test, or Wilcoxon signed rank test, with P < .05. RESULTS: Dogs were 6.5 ± 2.4 years old (mean ± SD) and had a body weight of 26.7 ± 5.2 kg. At T0, 32 (80%) joints were graded as having mild OA, 6 (15%) as moderate, and 2 (5%) as severe. No differences were found between groups at T0. Between the PCG and CG, the symmetry index showed significant improvements in the PCG from 8 days (P = .01) to 180 days (P = .01). Joint flexion also improved in the PCG up to 90 days (P < .05) and extension improved up to 180 days (P < .01). Several clinical metrology instrument scores also improved up to 90 to 180 days after treatment. In the CG, radiographic signs progressed, while the PCG showed some improved signs. In both groups, increasing body weight and age corresponded with worse clinical and laboratory findings. CONCLUSION: A single injection of platelet concentrate had a positive effect, lasting up to 6 months, on several clinical, imaging, and laboratory signs in a naturally occurring canine OA model. CLINICAL RELEVANCE: We characterized the effects of this platelet concentrate in dogs, considered the gold standard of the study of OA, with a group of working animals with similar high demands as athletes.

Thermographic imaging of police working dogs with bilateral naturally occurring hip osteoarthritis.

BACKGROUND: Digital thermal imaging is a physiologic, non-invasive, contactless, and non-radiating diagnostic tool that can assess a wide range of musculoskeletal conditions, including hip osteoarthritis (HOA). Fifty police working dogs were evaluated to compare the dorsoventral (DV) and lateral (LT) thermographic images in dogs with naturally occurring bilateral HOA. A DV, and left and right lateral LT images were obtained for each animal in six different moments. They were positioned standing in a symmetrical upright position for the DV view. Each image included the area from the last lumbar to the first coccygeal vertebrae. Each LT view was set with the greater trochanter in the centre of the image. Images were taken with a thermographic camera from a distance of 60 cm. Mean and maximal temperatures were recorded, analyzed with ANOVA, dependent samples t-test, and Spearman correlation, with P < 0.05. RESULTS: Nine hundred images were considered, collected from 30 males and 20 females, with a mean age of 6.5 ± 2.2 years and bodyweight of 26.7 ± 5.3 kg. The overall value recorded on the DV view was 25.3º ± 9.1 and 28.4º ± 2.8 on the lateral view. These were significantly different (P < 0.01) and with a low correlation (r = 0.10, P = 0.03). German Shepard dogs showed significantly lower values on all views than other breeds (P < 0.01), and heavier dogs had higher values on the lateral view. CONCLUSIONS: This is the first study that describes digital thermography's diagnostic use to evaluate working dogs with naturally occurring HOA, comparing two different views. Future studies should address each one's value in the diagnosis and response to treatment of this disease.

Operative Hysteroscopy Intravascular Absorption Syndrome: The Gynaecology's TURP Syndrome-A Case Report.

INTRODUCTION: Operative hysteroscopy intravascular absorption (OHIA) syndrome is caused by intravascular absorption of fluid distension/irrigation medium during hysteroscopy. There are very few reported cases of this syndrome using saline as irrigation fluid. The current report was on a case of severe OHIA syndrome that necessitated resuscitation in an intensive care unit (ICU). CASE PRESENTATION: A 41-year-old, 65-kg, smoker female patient was admitted for an endoscopic resection of submucous uterine myoma under general anesthesia using a laryngeal mask airway. In the 50th minute of the procedure, end-tidal CO2 dropped from 35 to 25 mmHg and pulse oximetry (SpO2) from 100% to 90%. Crackling sounds were heard from the base of the lungs. As a pulmonary oedema was suspected, the volume of irrigated saline was checked. A total of 4000 of the total 9000 mL of the saline had been absorbed into intravascular compartment. She developed a severe metabolic acidosis (pH 7.09) with severe hypokalemia (K+ 2.3 mEq/L), hypocalcaemia (Ca2+ 0.76 mEq/L), anemia (hemoglobin 5.3 g/dL), and hypothermia (tympanic temperature 33°C), as well as a generalized oedema with pulmonary and airway oedema. Due to airway oedema, she could only be intubated with a 6.5-mm tracheal tube. Resuscitation in the ICU was required. Electrolyte disturbances were corrected and furosemide was administered. She had a full recovery after 24 hours and 48 hours later, she was discharged. CONCLUSIONS: Absorption of the irrigation fluid can result in life-threatening fluid overload. Accurate fluid balancing and limiting the operation time may prevent such complications. Therefore, early diagnosis and treatment of this syndrome is emphasized.

Detection of Extravascular Trypanosoma Parasites by Fine Needle Aspiration.

Fine Needle Aspiration (FNA) is a routine diagnostic procedure essential to both medical and veterinary practices. It consists of the percutaneous aspiration of cells and/or microorganisms from palpable masses, organs or effusions (fluid accumulation in a body cavity) using a thin needle similar to the regular needle used for the venous puncture. The material collected by FNA is in general highly cellular, and the retrieved aspirate is then smeared, air dried, wet-fixed, stained and observed under a microscope. In the clinical context, FNA is an important diagnostic tool that serves as a guide to the appropriate therapeutic management. Because it is simple, fast, minimally invasive and requires limited investment in the laboratory and human resources, it is extensively used by veterinary practitioners, mostly in domestic, but also in farm animals. In studies using animal models, FNA has the advantage that it can be performed repeatedly in the same animal, enabling longitudinal studies through the collection of cells from tumors and organs/tissues over the course of the disease. In addition to routine microscopy, retrieved material can also be used for immunocytochemistry, electron microscopy, biochemical analysis, flow cytometry, molecular biology or in vitro assays. FNA has been used to identify the protozoan parasite Trypanosoma brucei in the gonads of infected mice, opening the possibility for a future diagnosis in cattle.

Evaluation of the Effect of Mesotherapy in the Management of Osteoarthritis-Related Pain in a Police Working Dog Using the Canine Brief Pain Inventory.

CASE DESCRIPTON: A 9-year-old, 33.4kg (73.63Lb) male entire drug detection Labrador Retriever Dog was presented with an history of constant lameness from the right thoracic limb, aggravated with exercise and work. CLINICAL FIDINGS: Clinical examination revealed mild signs of pain on the manipulation of the elbow joint, with reduced range of motion on the end feel of joint flexion and extension and crepitation. Radiographic examination of the right elbow joint revealed severe, chronic osteoarthritis, with osteophyte formation on the humeral epicondyles and articular margin of the distomedial humerus, with a narrowed joint space, and osteophytes on the proximal radius, proximomedial ulna, and anconeal process. TREATMENT AND OUTCOME: A solution comprised of a combination of lidocaine, thiocolchicoside, and piroxicam was prepared and applied around the right elbow joint. The animal was rested for 3 days and normal work load was introduced over a 5-day period. The CPBI was completed by the trainer before treatment (T0), 14 days (T1), 1 (T2), 2 (T3), 3 (T4), 4 (T5), 5 (T6), and 6 (T7) months after treatment. Following the mesotherapy session, pain score results consistently declined until the 3-month evaluation moment. At the 6-month follow-up evaluation, values have risen to near baseline values. No side effects were recorded. CLINICAL RELEVANCE: Mesotherapy produced significant reduction of pain score results, as measure by the CBPI, and may be a promising treatment option for canine osteoarthritis-related pain. Further studies are required.

Effect of an Oral Joint Supplement When Compared to Carprofen in the Management of Hip Osteoarthritis in Working Dogs.

The goal of this study was to evaluate the effectiveness of an oral joint supplement in working dogs with hip osteoarthritis compared with a positive control group (CG). Fifteen animals were divided in treatment group (TG, n = 10) and CG (n = 5). To TG a commercially available joint supplement, containing glucosamine HCl, chondroitin sulphate, and hyaluronic acid was given for 40 days and a 70-day course of a placebo, to be administered as if it was carprofen. The CG received carprofen for 70 days, and a placebo to be administered as the joint supplement. Response to treatment, measured by the canine brief pain inventory (CBPI) and the Hudson visual analog scale, was evaluated before treatment (T0), after 15 days (T1) and 1 (T2), 2 (T3), 3 (T4), 4 (T5), and 5 (T6) months. With CBPI, no differences were found in pain interference score and pain severity score between TG and CG throughout or when comparing results within groups. Individual results were considered successful in a maximal of three dogs of the TG by T3 (30%) and 1 in CG (25%). With Hudson visual analog scale, improvements where registered with individual results, for 40%-50% of the animals in TG and 60%-80% of cases in CG. The oral joint supplement and carprofen produced some improvements in individual scores but where unable to do so when overall results were considered. Each of these options may not be able, by itself, to fully address the demands of a working dog with joint disease and related pain.

Percepção dos familiares cuidadores informais de utentes hospitalizados no serviço de Medicina 1A com diagnóstico de AVC acerca das ajudas, apoios, ensinos / Perception of family informal caregivers of patients hospitalized in the Medicine 1A service with diagnosis of stroke about the help, support, teachings

A escolha do tema "Percepções dos familiares/cuidadores informais de utentes hospitalizados no Serviço de Medicina 1 A com diagnóstico de AVC acerca das ajudas/apoio/ensinos" deve-se ao facto do Acidente Vascular Cerebral (AVC) ser uma das patologias mais comuns da actualidade, resultando em grandes níveis de dependência dos doentes e em muita sobrecarga para os familiares. Este é um problema complexo, resultando na marginalização destes doentes, após a sua alta clínica, justificando uma actuação junto das famílias para a necessária modificação desta situação. Realizámos um estudo qualitativo, descritivo com características fenomenológicas, objectivando saber: se os familiares cuidadores de doentes vítimas de AVC têm ajuda de familiares ou das redes sociais para cuidar do seu familiar no domicílio; quais as doenças crónicas do doente; quais as dificuldades funcionais do doente; que ensinos foram realizados por parte dos profissionais de saúde para que exista uma continuidade de cuidados no domicílio; que dificuldades o cuidador informal sente ao cuidar do seu familiar doente; quais as implicações na sua vida familiar, social e profissional; que opiniões têm os participantes sobre o que pensam ser melhor para o seu familiar. O método de amostragem seguido foi o não probabilístico acidental por conveniência, num total de 10 cuidadoras informais. O método de recolha de dados foi a entrevista semi-estruturada. Concluímos que as cuidadoras informais são esposas e filhas. Na globalidade, não têm outro tipo de ajuda para cuidar o seu doente, a não ser a ajuda de outros familiares. Os ensinos realizados pelos profissionais de saúde foram: vestir o doente ou auxiliá-lo a trocar de roupa, levante, caminhar, transferências, alimentação e higiene, os quais foram ao encontro das suas maiores dificuldades. As entrevistadas sentem-se abaladas psicologicamente, resultante da angústia da situação actual e do futuro, ponderando algumas a necessidade de recorrer à institucionalização do seu doente.

The theme "Perceptions of family/ informal caregivers of patients hospitalized in the Medicine Service 1A due to a stroke diagnosis on the support/ help/ teachings" is due to the fact of Cerebrovascular Accident (CVA) is one of the most common pathologies today, resulting in high levels of patient dependency and a lot of overhead for family members. This is a complex problem, resulting in the marginalization of these patients after their clinical high, justifying an action with the families for the necessary modification of this situation. We conducted a qualitative, descriptive study with phenomenological features, aiming to know: whether the family caregivers of stroke patients have help from family or social networks to care for them at home; the chronic diseases of the patient; the functional difficulties of the patient; the teachings conducted by health professionals so that there is a continuity of care at home; the difficulties felt by informal caregiver when taking care of the sick family member; the implications for family, social and professional life; participants' opinions about what is best for their sick family member. The sampling method followed was a not accidental probabilistic by convenience, for a total of 10 informal caregivers. The method of data collection was a semi-structured interview. We conclude that informal caregivers are wives and daughters. Overall, they have no other help to care for their patient, unless the help of other family members. The teachings conducted by health professionals were: dress the patient or help him changing clothes, to get up, to walk, transfers, food and hygiene. Interviewees feel psychologically shaken, resulting from the distress of the current situation and future, some weighing the need for the institutionalization of his sick family member.

An integrated strategy to identify key genes in almond adventitious shoot regeneration.

Plant genetic transformation usually depends on efficient adventitious regeneration systems. In almond (Prunus dulcis Mill.), regeneration of transgenic adventitious shoots was achieved but with low efficiency. Histological studies identified two main stages of organogenesis in almond explants that were induced for adventitious shoot regeneration; a dedifferentiation stage (early) and a shoot initiation stage (late). Histological observation revealed that the limitation in the recovery of transformed shoots is primarily a function of the low organogenic competence of the transformed tissues rather than transformation efficiency. To identify key genes involved in organogenesis, shoot-induced leaves and suppression-subtractive hybridization were used, to build a cDNA library from each organogenic stage. cDNA clones from both libraries were randomly picked, PCR-amplified, and arrayed on glass slides. For transcript profiling, microarray hybridization was performed using cDNA pools from both the early and the late stages. Statistically significant differential expression was found for 128 cDNA clones (58 early, and 70 late), representing 92 unique gene functions. Genes encoding proteins related to protein synthesis and processing and nitrogen and carbon metabolism were differentially expressed in the early stage, whilst genes encoding proteins involved in plant cell rescue and defence and interaction with the environment were mostly found in the late stage. The LTP/alpha-amylase inhibitor/trypsin gene was more strongly expressed at an early stage, as confirmed by quantitative RT-PCR, while a gibberellic acid stimulated protein gene seems to be a good marker for the late stage. These results are discussed on the basis of the putative roles of the annotated differentially regulated genes in almond organogenesis.