Evaluation of a Low-cost Renal Simulator for the Diagnostic Ultrasound Training of Veterinary Medicine Students.

The acquisition of ultrasound diagnostic skills via training is important for undergraduate veterinary medical students. Unfortunately, commercial simulators are costly, which limits their use and makes it necessary to develop low-cost simulators for training purposes. The aim of this study was to evaluate the efficacy of an easily constructed, low-cost, high-fidelity renal simulator for use in diagnostic ultrasound training of veterinary medical students. To construct the simulators, donated cat kidneys were embedded in a prepared agar matrix. The echogenicity of ultrasound scans obtained on the kidney simulator was assessed by the subject lecturers and compared with images acquired during clinical routine diagnostic procedures. Five students with no prior experience of the technique, under the direct supervision of a lecturer, performed five B-mode ultrasound examinations of the renal structure of the simulator. The structure of the kidney was assessed, and its length, width and thickness were measured. Three lecturers performed the same procedure as the students, and their results were used for comparison. Appropriate anatomical and ultrasonographic realism was achieved for each of the three layers of the kidney (cortex, medulla and pelvis), and similar pixel values were obtained with the simulator model and actual clinical diagnostic ultrasounds. In addition, the kidney dimensions acquired by the students were consistent with those acquired by the lecturers. Thus, the proposed kidney simulator can be used for the training of veterinary medicine students in ultrasonographic diagnostic techniques.

Effect of intermittent induced aeration on nitrogen removal and denitrifying-bacterial community structure in Cork and gravel vertical flow pilot-scale treatment wetlands.

In this work, we have evaluated the impact of intermittent induced aeration in total nitrogen (TN), ammonia (NH4-N) and nitrate-nitrogen (NO3-N) removal in four pilot-scale vertical flow constructed wetlands (VFCW) (two aerated two non-aerated) using cork by-product or gravel as the filter material and planted with Phragmites australis. Both aerated and non-aerated systems achieved high COD and BOD5 elimination rates (≥ 90%) at the end of the 5-month test period. However, the aerated systems presented maximal COD and BOD5 removal from the third month of operation onwards since air supply favored the oxidative bioprocesses occurring within the wetlands. Cork and gravel aerated VFCW also proved to be more efficient (p < 0.05) in NO3-N removal than the non-aerated systems and this upgraded performance was correlated with a significant higher relative abundance of the nirS gene. The aerated systems also showed a slightly improved NH4-N removal. Noticeably, cork VFCW showed higher TN removal mean values (∼35%) than gravel wetlands (27-28%) regardless aeration. Moreover, cork VFCW showed higher relative abundance of the nosZ gene. Our results demonstrated a better nitrogen elimination for the aerated cork pilot-scale VFCW, and this behavior was correlated with a higher abundance of both nirS and nosZ, two of the key functional genes involved in nitrogen metabolism.

RADIOGRAPHIC ANATOMY OF THE FORELIMB IN THE NORTHERN TAMANDUA (TAMANDUA MEXICANA).

The thoracic limb anatomy of anteaters in the family Myrmecophagidae is specialized for accessing termite and ant nests and for defense purposes. In the case of the northern tamandua (Tamandua mexicana), the forelimbs are also adapted for arboreal and terrestrial locomotion. Unfortunately, this species faces many conservation threats, such as habitat loss and traffic accidents, and injured individuals are frequently taken to wildlife rehabilitation centers. However, lack of knowledge of the radiographic osteoanatomy of this species may prevent appropriate management of injuries and thereby reduce the chances of successful release and survival. In order to fill this knowledge gap, this article describes for the first time the radiographic anatomy of the thoracic limb of the northern tamandua using four standard views and one additional view. The additional orthogonal view helps visualize structures, such as the hamatus process and the sesamoid bone, that are otherwise difficult to visualize due to the natural forearm position of anteaters. Additionally, some fractures and physeal growth plates were identified in one juvenile individual. Further radiographic investigations should be conducted on anteaters to provide more tools for diagnosis, treatment, and rehabilitation of these animals.

Surgical Extraction of Viable Hen ( Gallus gallus domesticus) Follicles for In Vitro Fertilization.

Assisted reproduction techniques in birds have been developed for zootechnical purposes and have been adapted for use in conservation of wild bird species. To develop a technique for obtaining follicles in live hens, 5 Rhode Island red hens ( Gallus gallus domesticus) were anesthetized, and abdominal ultrasound was performed to confirm the presence of ovarian follicles. A left celiotomy then was performed to obtain follicles in different stages of maturation for in vitro fertilization. The follicles were located by digital exploration, then extracted by isolating each follicle with the index finger of each hand, holding it by the stigma, and then applying slight traction towards the exterior of the coelomic cavity until the follicle separated from the ovary. In total, 18 of 30 (60%) follicles obtained were suitable for in vitro fertilization, but only 3 (16%) were fertilized successfully. All birds recovered from the procedure and remained in good condition postoperatively. Perfecting assisted reproduction technique holds potential benefits for determining sex of embryos by blastomeres sexing, supporting the conservation efforts of avian species, and benefiting research areas, such as genetic and biopharmaceutical research.

Development of a bone nonunion in a noncritical segmental tibia defect model in sheep utilizing interlocking nail as an internal fixation system.

BACKGROUND: The bone nonunion is an important complication of bone fracture repair. The existing models developed on small animal species prevent using osteosynthesis materials designed to be implanted in human bones. The goal of this study was to develop a nonunion process in a noncritical segmental tibial defect in sheep, a species analogous in size to humans. MATERIALS AND METHODS: The animals were divided into two groups of four animals each. In Group 1 (experimental), the defect was created by surgically stripping the periosteum from the edges of a distal tibial osteotomy, keeping the edges 5 mm apart, and placing an incomplete O-shaped silicone ring in the gap. Group 2 (control) was intervened with a simple fracture at the distal end of the tibia. In both groups an interlocking nail was used as a fixation system. Over 8 wk after surgery, radiographs and histologic and histomorphometric analyses were performed. RESULTS: The control group showed a typical bone repair process. In contrast, the experimental group showed a fracture line with rounded edges and a scarce callus formation. The bone callus showed reduced amount of bone formation and large content of fibrous tissue (P=0.001). CONCLUSIONS: These results indicate that our model developed an atrophic nonunion in sheep, a species having multiple similarities to humans, such as weight, size, bone structure, and bone remodeling process.

CD24+CD44+CD54+EpCAM+ gastric cancer stem cells predict tumor progression and metastasis: clinical and experimental evidence.

BACKGROUND: Gastric cancer (GC) is a leading cause of cancer-related deaths worldwide. Specific and thorough identification of cancer cell subsets with higher tumorigenicity and chemoresistance, such as cancer stem cells (CSCs), could lead to the development of new and promising therapeutic targets. For better CSC identification, a complete or extended surface marker phenotype is needed to provide increased specificity for new cell targeting approaches. Our goal is to identify and characterize a putative extended phenotype for CSCs derived from patients with GC before treatment, as well as to evaluate its clinical value. In addition, we aim to ensure that cells with this phenotype have stemness and self-renewal capabilities. METHODS: This is a cohort study including 127 treatment-naïve patients with GC who attended the Instituto Nacional de Cancerología. Multiparametric flow cytometry analysis was performed to determine the extended phenotype of cells derived from gastric biopsies. The tumorigenic capability of cells identified in patients was assessed in a zebrafish model. RESULTS: CD24+CD44+CD54+EpCAM+ cells were present in all treatment-naïve patients included, with a median abundance of 1.16% (0.57-1.89%). The percentage of CD24+CD44+CD54+EpCAM+ cells was categorized as high or low using 1.19% as the cutoff for the CD24+CD44+CD54+EpCAM+ cell subset. Additionally, a higher TNM stage correlated with a higher percentage of CD24+CD44+CD54+EpCAM+ cells (Rho coefficient 0.369; p < 0.0001). We also demonstrated that a higher percentage of CD24+CD44+CD54+EpCAM+ cells was positively associated with metastasis. The metastatic potential of these cells was confirmed in a zebrafish model. Ultimately, under our conditions, we conclude that CD24+CD44+CD54+EpCAM+ cells are true gastric cancer stem cells (GCSCs). CONCLUSION: The CD24+CD44+CD54+EpCAM+ cells present in tissue samples from patients are true GCSCs. This extended phenotype results in better and more specific characterization of these highly tumorigenic cells. The relative quantification of CD24+CD44+CD54+EpCAM+ cells has potential clinical value, as these cells are associated with metastatic disease, making their presence an additional prognostic marker and possibly a target for the design of new antineoplastic treatments in the era of precision oncology. Overall, the extended CD24+CD44+CD54+EpCAM+ phenotype of GCSCs could support their isolation for the study of their stemness mechanisms, leading to the identification of better molecular targets for the development of both new therapeutic approaches such as oncoimmunotherapy and new diagnostic and clinical prognostic strategies for GC.

Determination of (fluoro)quinolones in environmental water using online preconcentration with column switching linked to large sample volumes and fluorescence detection.

An analytical method based on online enrichment using coupled-column liquid chromatography with fluorescence detection has been developed to determine marbofloxacin, ciprofloxacin, danofloxacin, enrofloxacin, norfloxacin, lomefloxacin, oxolinic acid, and nalidixic acid at trace levels in surface water. The sample containing the pharmaceuticals was pumped through a short C18 column in such a way that the analytes were retained on the column, whereas polar interferences, eluting at the first of the chromatogram, were discarded to waste. Then, the analytes were transferred by the chromatographic mobile phase to a second C18 analytical column, where they were separated following a conventional chromatography. The optimized approach allowed to preconcentrate 15 mL of sample volume adjusted at acid pH with phosphoric acid and modified with 5% of methanol, at a flow rate of 1.5 mL/min in 10 min. R(2) values were between 0.994 and 0.998, detection and quantitation limits ranged between 0.001 and 0.080 and between 0.002 and 0.100 µg/L, respectively, and the interday precision was below 9.8%. Recoveries in three different surface water samples, spiked at concentration levels between 0.002 and 0.500 µg/L (n = 3 for each spiking level), ranged from 82.1 to 125.8% with the relative standard deviation lower than 12%.

Bispecific CAR T Cells against EpCAM and Inducible ICAM-1 Overcome Antigen Heterogeneity and Generate Superior Antitumor Responses.

Adoptive transfer of chimeric antigen receptor (CAR) T cells has demonstrated unparalleled responses in hematologic cancers, yet antigen escape and tumor relapse occur frequently. CAR T-cell therapy for patients with solid tumors faces even greater challenges due to the immunosuppressive tumor environment and antigen heterogeneity. Here, we developed a bispecific CAR to simultaneously target epithelial cell adhesion molecule (EpCAM) and intercellular adhesion molecule 1 (ICAM-1) to overcome antigen escape and to improve the durability of tumor responses. ICAM-1 is an adhesion molecule inducible by inflammatory cytokines and elevated in many types of tumors. Our study demonstrates superior efficacy of bispecific CAR T cells compared with CAR T cells targeting a single primary antigen. Bispecific CAR T achieved more durable antitumor responses in tumor models with either homogenous or heterogenous expression of EpCAM. We also showed that the activation of CAR T cells against EpCAM in tumors led to upregulation of ICAM-1, which rendered tumors more susceptible to ICAM-1 targeting by bispecific CAR T cells. Our strategy of additional targeting of ICAM-1 may have broad applications in augmenting the activity of CAR T cells against primary tumor antigens that are prone to antigen loss or downregulation.

ALDH as a Stem Cell Marker in Solid Tumors.

Aldehyde dehydrogenase (ALDH) is an enzyme that participates in important cellular mechanisms as aldehyde detoxification and retinoic acid synthesis; moreover, ALDH activity is involved in drug resistance, a characteristic of cancer stem cells (CSCs). Even though ALDH is found in stem cells, CSCs and progenitor cells, this enzyme has been successfully used to identify and isolate cell populations with CSC properties from several tumor origins. ALDH is allegedly involved in cell differentiation through its product, retinoic acid. However, direct or indirect ALDH inhibition, using specific inhibitors or retinoic acid, has shown a reduction in ALDH activity, along with the loss of stem cell traits, reduction of cell proliferation, invasion, and drug sensitization. For these reasons, ALDH and retinoic acid are promising therapeutic targets. This review summarizes the current evidence for ALDH as a CSCs marker in solid tumors, as well as current knowledge about the functional roles of ALDH in CSCs. We discuss the controversy of ALDH activity to maintain CSC stemness, or conversely, to promote cell differentiation. Finally, we review the advances in using ALDH inhibitors as anti-cancer drugs.

Microbial nitrate removal efficiency in groundwater polluted from agricultural activities with hybrid cork treatment wetlands.

Agricultural practices have raised the level of nutrients reaching aquifers. In Europe, nitrate pollution is considered as one of the main threats for the quality of groundwater in agricultural areas. Treatment wetlands (TWs), also known as Constructed Wetlands, are used for groundwater treatment in areas with an important concentration of nitrogen compounds; total nitrogen removal depends on the type and operation scheme. Cork by-product from the industry has shown clear adsorbent properties to remove organic pollutants. The work is focused on the characterization of microbial communities involved in the nitrate­nitrogen removal process in groundwater polluted from agricultural activities. The experimental design allowed the comparison of nitrate removal efficiency depending on the filter media material, cork by-product or gravel, used in two hybrid TWs (a vertical flow cell followed by a horizontal subsurface flow cell), installed in areas close to two irrigated agricultural plots at the Lleida plain area (Spain). Both physicochemical and microbial results were consistent and confirm the nitrate removal efficiency using cork as a filter media. A significant (p = 0.0025) higher removal in Bellvís TW using cork compared with the Vilanova de la Barca gravel system was observed, achieving a removal rate from 80 to 99% compared to the 5-46%, respectively. Regarding the community composition of the two different TWs, microorganisms were mainly related to the phylum Proteobacteria, and included members found to be key players in the nitrogen cycle, such as ammonia and nitrite oxidizers, as well as denitrifiers. Also, the group Bacteroidetes turns to be another abundant phylum from our bacterial dataset, whose members are suggested to be strongly involved in denitrification processes. Some groups showed to prevail depending on the type of media (cork or gravel); Firmicutes and Delta and Epsilonproteobacteria had a significant higher abundance in the TW with cork, while Acidobacteria and Planctomyces were prevalent in gravel. Therefore, cork could be an alternative material used by treatment wetlands to minimize the impact in the environment caused by nitrogen pollution in groundwater bodies.

Fracturas longitudinales de sacro en Zona III Dennis, ¿Qué sabemos al respecto? / Longitudinal sacral fractures Dennis zone III, what do we know about them?

Objetivo: Las fracturas pelvicoacetabulares constituyen un tipo de fractura asociada a traumatismos de alta energía, que no sólo ponen en riesgo la vida del paciente, sino que además se asocian con un alto porcentaje de secuelas. Las fracturas de sacro longitudinales en línea media constituyen un sub-grupo raro de fracturas de sacro de la Zona III de Dennis que se revisan en este artículo a propósito de un caso atendido en nuestro centro. Material y Métodos: Varón de 63 años que tras un accidente de motocicleta presentó una fractura pélvicoacetabular por compresión anteroposterior (APC) III de Young-Burguess con una fractura vertical de sacro en la zona III de Dennis y diástasis de la sínfisis púbica de 5 cm. Resultados: Actualmente el paciente se encuentra caminando con bastones y con parestesias en fase de remisión. Conclusiones: La incidencia de lesiones neurológicas en fracturas de pelvis fluctúa entre el 3% y el 15 % y aumenta cuanto mayor sea la energía o el desplazamiento. Las fracturas de pelvis suponen un reto diagnóstico y terapeútico para el traumatólogo, debido tanto a la mortalidad que presentan como al gran número de secuelas que pueden acontecer (disfunciones intestinales, vesicales, sexuales, dolor neuropático)

Aim: Pelvic-acetabular fractures are a kind of fracture associated with high-energy trauma, which not only endanger the life of the patient but also are associated with a high percentage of sequelae. The longitudinal midline sacrum fractures constitute a rare subgroup of sacrum fractures of Dennis’ Zone III, which are reviewed in this article in relation to a case attended in our center. Material and Methods: A 63-year-old male presented a Young-Burguess anteroposterior (APC) III pelvic-acetabular fracture (APC) after a motorcycle accident with a vertical sacrum fracture in Dennis area III and pubic symphysis diastasis of 5 cm. Results: Currently the patient is walking with sticks and paresthesias getting better. Conclusions:The incidence of neurological injuries in pelvic fractures ranges between 3% and 15% and increases the higher the energy or displacement. Pelvic fractures pose a diagnostic as well as a therapeutic challenge for the Orthopaedics surgeons, due both to the mortality they present and to the large number of sequelae that may occur (intestinal, bladder, sexual dysfunction, neuropathic pain)

Power Doppler ultrasonography assessment of entheses in spondyloarthropathies: response to therapy of entheseal abnormalities.

OBJECTIVE: To investigate the response to therapy of entheseal abnormalities assessed with power Doppler (PD) ultrasound (US) in spondyloarthropathies (SpA). METHODS: A total of 327 patients with active SpA who were starting anti-tumor necrosis factor (TNF) therapy were prospectively recruited at 35 Spanish centers. A PDUS examination of 14 peripheral entheses was performed by the same investigator in each center at baseline and at 6 months. The following elementary lesions were assessed at each enthesis (presence/absence): morphologic abnormalities (hypoechogenicity and/or thickening), entheseal calcific deposits, cortical abnormalities (bone erosion and/or proliferation), adjacent bursitis and intraenthesis and perienthesis (tendon body and/or bursa) PD signal. Response to therapy of each elementary lesion was assessed by calculating change in the cumulative presence from baseline to 6 months. Intraobserver reliability of PDUS was evaluated by blindly assessing the stored baseline images 3 months after the real-time examination. RESULTS: Complete data were obtained on 197 patients who received anti-TNF therapy for 6 months. In 91.4% of the patients there were gray-scale or PD elementary lesions at baseline and at 6 months. Cumulative entheseal morphologic abnormalities, intraenthesis PD, perienthesis PD, and bursitis showed a significant decrease from baseline to 6 months (p < 0.05). There was high intraobserver reliability for all elementary lesions (interclass correlation coefficient > 0.90, p < 0.0005). CONCLUSION: Entheseal morphologic abnormalities, PD signal, and bursitis were US abnormalities that were responsive to anti-TNF therapy in SpA. PDUS can be a reproducible method for multicenter monitoring of therapeutic response in enthesitis of SpA.