RESUMO
The study aimed to compare the quality of perioperative analgesia, the motor block duration, and the effects on main cardiovascular parameters of dexmedetomidine (1 µg/kg/nerve block) or magnesium sulphate (2 mg/kg/nerve block) as adjuvants to 0.3% ropivacaine for sciatic and saphenous nerves block in dogs undergoing tibial plateau leveling osteotomy (TPLO). Dogs randomly received perineural dexmedetomidine-ropivacaine (D group), magnesium sulphate-ropivacaine (M group), or ropivacaine (C group). Fentanyl was administered in case of intraoperative nociception. Postoperative pain was assessed using the Short Form-Glasgow Composite Measure Pain Scale (SF-GCMPS) and VAS scale. The duration of motor blockade and intra- and postoperative cardiovascular parameters were also recorded. Group M required significantly more fentanyl than D group (p = 0.04). Group M had a significantly higher SF-GCMPS score than group C at 4 (p = 0.002) and 5 h after extubation (p = 0.01), and a significantly higher VAS score than group D at 3 h after extubation (p = 0.03), and at 4 h if compared to group C (p = 0.009). No significant differences regarding the duration of motor blockade were detected between groups (p = 0.07). The heart rate was significantly lower in group D than in M and C groups intraoperatively and during the first 1.5 h post extubation. The addition of dexmedetomidine or magnesium sulphate as adjuvants to perineural ropivacaine did not improve the quality of perioperative analgesia and did not prolong the motor blockade in dogs undergoing sciatic and saphenous nerves block for TPLO surgery.
Assuntos
Dexmedetomidina , Sulfato de Magnésio , Bloqueio Nervoso , Osteotomia , Dor Pós-Operatória , Ropivacaina , Animais , Cães , Dexmedetomidina/administração & dosagem , Dexmedetomidina/farmacologia , Ropivacaina/administração & dosagem , Ropivacaina/farmacologia , Sulfato de Magnésio/farmacologia , Sulfato de Magnésio/administração & dosagem , Osteotomia/veterinária , Dor Pós-Operatória/veterinária , Dor Pós-Operatória/tratamento farmacológico , Dor Pós-Operatória/prevenção & controle , Masculino , Feminino , Bloqueio Nervoso/veterinária , Anestésicos Locais/administração & dosagem , Anestésicos Locais/farmacologia , Tíbia/cirurgiaRESUMO
During tibial plateau leveling osteotomy (TPLO), the laceration of the cranial tibial artery (LCTA) may occur, and the ligation of the cranial tibial artery might lead to impaired blood supply to the osteotomy site. The present case-control study aimed to evaluate the effect of LCTA on TPLO healing and the occurrence of perioperative complications. The incidence and predisposing factors to LCTA were also investigated. Fourteen cases experiencing LCTA were retrospectively enrolled from medical records of two veterinary teaching hospitals (LCTA group), whereas 28 randomly selected TPLOs that did not experience LCTA were included in the control group. Signalment data, proximal tibial epiphysis conformation, osteotomy features, perioperative complications, and bone healing were compared between the two groups. Bone healing was evaluated using the modified radiographic union scale for tibial fracture and the visual analog scale. The mean incidence was 9.6%. Bodyweight was significantly higher in the LCTA group compared to the control group (P = 0.009). Dogs belonging to the LCTA groups were significantly younger (P = 0.01). Intraoperative hypotension was significantly overreported in the LCTA group (P = 0.0001). None of the other variables differed significantly between the two groups. Dogs' size seems to be a predisposing factor, with dogs weighing >15 kg having 22 times more chance of experiencing LCTA. Due to the well-developed collateral blood supply of the canine hindlimb, LCTA and the closure of the cranial tibial artery did not appear to delay the radiographic bone healing or affect the incidence of perioperative complications.
Assuntos
Osteotomia , Tíbia , Artérias da Tíbia , Animais , Cães , Osteotomia/veterinária , Feminino , Masculino , Estudos de Casos e Controles , Estudos Retrospectivos , Tíbia/cirurgia , Tíbia/irrigação sanguínea , Artérias da Tíbia/diagnóstico por imagem , Artérias da Tíbia/lesões , Artérias da Tíbia/cirurgia , Complicações Pós-Operatórias/veterinária , Complicações Pós-Operatórias/etiologia , Doenças do Cão/cirurgia , Doenças do Cão/diagnóstico por imagem , Consolidação da Fratura , Lacerações/veterinária , Lacerações/cirurgiaRESUMO
BACKGROUND: Reports describing sciatic nerve injuries (SNI) and their outcome are scarce in veterinary medicine. HYPOTHESIS: Describe the causes of traumatic and iatrogenic SNI and evaluate which clinical and electrodiagnostic findings predict outcome. ANIMALS: Thirty-eight dogs and 10 cats with confirmed SNI referred for neurologic and electrodiagnostic evaluation. METHODS: Clinical and electrodiagnostic examination results, including electromyography (EMG), motor nerve conduction studies, muscle-evoked potential (MEP), F-waves, sensory nerve conduction studies, and cord dorsum potential (CDP), were retrospectively evaluated. Quality of life (QoL) was assessed based on owner interviews. RESULTS: Surgery (42%) and trauma (33%) were the most common causes of SNI; in dogs, 24% were caused by bites from wild boars. Ability to flex and extend the tarsus was significantly associated with positive outcome in dogs. Mean time from onset of clinical signs until electrodiagnostic evaluation was 67 ± 65 (range, 7-300) days and 65 ± 108 (range, 7-365) days for dogs and cats, respectively. A cut-off amplitude of 1.45 mV for compound motor action potentials (CMAP) was predictive of positive outcome in dogs (P = .01), with sensitivity of 58% and specificity of 100%. CONCLUSIONS AND CLINICAL IMPORTANCE: Clinical motor function predicts recovery better than sensory function. Electrodiagnostic findings also may play a role in predicting the outcome of SNI. Application of the proposed CMAP cut-off amplitude may assist clinicians in shortening the time to reassessment or for earlier suggestion of salvage procedures. Owners perceived a good quality of life (QoL), even in cases of hindlimb amputation.
Assuntos
Doenças do Cão , Eletromiografia , Nervo Isquiático , Animais , Cães , Gatos , Nervo Isquiático/lesões , Masculino , Feminino , Estudos Retrospectivos , Doenças do Cão/diagnóstico , Doenças do Cão/fisiopatologia , Eletromiografia/veterinária , Doenças do Gato/diagnóstico , Doenças do Gato/fisiopatologia , Qualidade de Vida , Eletrodiagnóstico/veterinária , Neuropatia Ciática/veterinária , Neuropatia Ciática/diagnóstico , Neuropatia Ciática/fisiopatologia , Doença Iatrogênica/veterinária , Condução Nervosa/fisiologiaRESUMO
Cardiovascular changes have been reported in late pregnancy in mares. However, there are no data on changes in peripheral blood flow. Doppler ultrasound represents a sensitive method for assessing the blood flow directed to the hoof. The aims of this study were to evaluate the blood flow parameters of the lateral palmar digital artery (LPDA) in pregnant mares and to assess intra- and interrater agreement between two observers with different levels of experience. The LPDAs of pregnant Italian Standardbred mares were examined. The vessels were located with B-mode ultrasound and analyzed with color and pulsed wave Doppler. The following parameters were recorded by the operators: heart rate (HR), peak systolic velocity (PSV), end-diastolic velocity (EDV), and resistive index (RI). Measurements were performed between 2 and 3 months of gestation (T1), in the last month of pregnancy (T2) and a week after delivery (T3). Seventeen mares aged 3-18 years met the inclusion criteria. Ultrasound examinations of the LPDA were subjectively easy to perform and well tolerated by the mares. Interrater and intrarater agreement were good and moderate, respectively. The HR was higher at T2 than at T1 and T3. The PSV and RI changed significantly during pregnancy, with higher values at T2 and T3, whereas the EDV remained unchanged throughout the examination. Doppler examination showed that peripheral flow changes were present in mares in late pregnancy. However, the persistence of higher values after delivery invites further investigation to assess the correlation between metabolic/endocrine changes related to pregnancy and Doppler parameters.
Assuntos
Prenhez , Animais , Cavalos , Feminino , Gravidez , Ultrassonografia Doppler/veterinária , Artérias/diagnóstico por imagem , Velocidade do Fluxo Sanguíneo/veterináriaRESUMO
BACKGROUND: The therapeutic role and prognostic relevance of lymphadenectomy in mast cell tumor (MCT) has historically been evaluated on regional rather than sentinel lymph nodes. HYPOTHESIS/OBJECTIVES: To update information about the association of histological nodal (HN) classes with clinical outcome in dogs with MCT after tumor excision and extirpation of normal-sized sentinel nodes (SLN) guided by radiopharmaceutical. ANIMALS: Ninety-four dogs with histologically-confirmed treatment-naïve MCT (71 cutaneous, 22 subcutaneous and 1 conjunctival MCT) were included if without: distant metastases, lymphadenomegaly, concurrent mixed cutaneous, and subcutaneous MCT. METHODS: This was a monoistitutional cohort study. Tumors characteristics were retrieved and SLNs were classified according to Weishaar's system. Incidence of MCT-related events (local, nodal, distant relapse), de novo MCT or other tumors and death (MCT-related and non-MCT-related), were recorded. Incidence curves were compared among the HN classes. RESULTS: Twenty-seven dogs had HN0, 19 HN1, 37 HN2, and 11 HN3 SLN. Thirteen (2 HN0, 4 HN2, and 7 HN3) received adjuvant chemotherapies. Kiupel high grade, increasing number of SLN and lymphocentrums were associated with higher HN classes. Five dogs died for MCT-related causes: 1 low-grade (HN0) and 1 subcutaneous (HN3) had a local relapse, 2 high-grade had distant relapse (HN3-HN0) and 1 dog developed disease progression from a de novo subcutaneous MCT. No nodal relapse was registered. Fourteen dogs developed de novo MCTs. CONCLUSION/DISCUSSION: Low grade/low-risk MCT with nonpalpable and normal sized SLN have a favorable outcome independently from the HN. Result should be considered strictly related to the successful SLN detection guided pre- and intraoperative by radiopharmaceutical markers.
Assuntos
Doenças do Cão , Metástase Linfática , Linfonodo Sentinela , Animais , Cães , Doenças do Cão/patologia , Feminino , Masculino , Metástase Linfática/patologia , Linfonodo Sentinela/patologia , Excisão de Linfonodo/veterinária , Estudos de Coortes , Mastocitoma/veterinária , Mastocitoma/patologia , Sarcoma de Mastócitos/veterinária , Sarcoma de Mastócitos/patologia , Resultado do TratamentoRESUMO
BACKGROUND: Inflammatory myopathy and perivasculitis have been recently described in horses with chronic equine piroplasmosis (EP). These alterations may be linked to poor performances. The aims of this study were to evaluate the prevalence for EP in clinically healthy Italian Standardbred (IS) racehorses and to compare laboratory parameters and performance metrics between positive and negative horses. Real-time PCR was applied for the detection of T. equi and B. caballi positivity. Haematology parameters, blood chemistry results, subjective muscle mass scores, and performance metrics were compared between PCR-positive and -negative horses. RESULTS: This cross-sectional study included 120 well-trained IS racehorses and was performed over a two-years period. The prevalence of T. equi was 36.3%, whereas all samples were negative for B. caballi. Red blood cells count, haemoglobin concentration, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase activities were significantly higher in PCR-positive horses, whereas blood urea nitrogen, globulin concentration and globulin-to-albumin ratio were significantly lower in PCR-positive horses compared to PCR-negative ones. Nonetheless, all values fell within the physiological range. The best racing time, which was selected as the most representative of the performance metrics at the principal component analysis, was not affected by PCR positivity, the muscle mass score or the training yard. The best racing time was significantly better in horses with a mild or no signs of muscular atrophy, within the PCR-positive group. The muscle mass score was associated with the training yard in PCR-negative horses. CONCLUSIONS: Prevalence of T. equi was high in IS racehorses in southern Italy. The absence of obvious changes in haematological and biochemical parameters, as well as performance metrics in positive horses, highlights the need for specific diagnostic tests to identify chronically infected horses.
Assuntos
Globulinas , Theileria , Animais , Cavalos , Estudos Transversais , Theileria/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Itália/epidemiologiaRESUMO
The management of unowned cats is an emerging problem, with public institutions and citizens' concerns regarding their care and arrangement. Little is known regarding the outcome of traumatic orthopedic injuries in these patients. Indeed, complete functional recovery (CFR) should be the goal of treatment for return to their original location or adoption. The aim was to identify clinical factors influencing CFR in traumatized unowned cats with orthopedic lesions. This category of cats referred by the veterinary public service over three years was enrolled. Various clinical variables were retrospectively collected from the medical records and evaluated by nominal logistic analysis. Forty-eight unowned cats were enrolled, with a median estimated age of 24 (1-180) months and a body weight of 3 (0.7-5) kg. Thirty-four (71%) patients reached CFR. Estimated age, body weight, time from trauma to therapeutic intervention, spine involvement, presence of comorbidities, hospitalization time, and the radiographic score results were significantly associated with CFR. A longer time to therapeutic intervention seemed to be associated with a better outcome. Probably, cats severely traumatized did not live long enough to be evaluated and treated. Lighter cats experienced more severe consequences following blunt trauma. Younger and lighter cats bore a higher risk of panleukopenia-related death.
RESUMO
Methylene Blue (MB) is combined with radiopharmaceutical for intraoperative sentinel lymph node (SLN) mapping, but its role during SLN extirpation has not been investigated yet in veterinary medicine. The aim of this study was to assess whether MB increased surgical detection of SLN beyond the use of intraoperative gamma-probe (IGP) alone in clinically node-negative dogs with mast cell tumors (MCTs) following the detection of sentinel lymphocentrums (SLCs) via preoperative planar lymphoscintigraphy. Dogs enrolled underwent MCT excision and SLC exploration guided by both MB and IGP. Data recorded for each SLN were staining (blue/non-blue), radioactivity (hot/non-hot), and histopathological status (HN0-1 vs. HN2-3). A total of 103 dogs bearing 80 cutaneous, 35 subcutaneous, and 1 mucocutaneous MCTs were included; 140 SLCs were explored, for a total of 196 SLNs removed. Associating MB with IGP raised the SLNs detection rate from 90% to 95%. A total of 44% of SLNs were metastatic: 86% were blue/hot, 7% were only blue, 5% were only hot, and 2% were non-blue/non-hot. All HN3 SLNs were hot. Combining MB with IGP can increase the rate of SLN detection in dogs with MCTs; nonetheless, all lymph nodes identified during dissection should be removed, as they might be unstained but metastatic.
RESUMO
The detection of subtle changes in the pituitary dimensions has relevant clinical implications. In cats, a few studies have established the cut-off values of the pituitary gland's dimensions using small and inhomogeneous samples. The aims of this study were: to determine by computed tomography (CT) the pituitary linear dimensions and the pituitary-to-brain (P:B) ratio in a sample of domestic short-haired (DSH) cats; to assess the effects of sex, age, and weight on pituitary dimensions; and to evaluate the inter- and intra-observer agreement for such measurements. All skull CTs of DSH cats performed over four years using a multidetector CT and a standardized protocol were retrospectively reviewed. The exclusion criteria were: clinical, laboratory, or CT alterations of the pituitary gland, brain diseases, fractures of the neurocranium, and diabetes. The pituitary dimensions and brain area were assessed by two different observers using multiplanar reconstructions and automated segmentation tools. Fifty-one cats were included in the final sample. The intraclass correlation coefficients for intra- and inter-observer reliability were good/excellent, and moderate/good, respectively. No differences between sexes were detected, and negligible correlations were found between age and weight. According to this study, a pituitary gland with a height > 4 mm or a P:B ratio > 0.49 mm should be considered enlarged.
RESUMO
BACKGROUND: Hypoxic tumor microenvironment (TME) contributes to the onset of many aspects of the cancer biology associated to the resistance to conventional therapies. Hypoxia is a common characteristic and negative prognostic factor in the head and neck squamous carcinomas (HNSCC) and is correlated with aggressive and invasive phenotype as well as with failure to chemo- and radio-therapies. The carbonic anhydrase isoenzymes IX and XII (CA IX/XII), regulators of extra and intracellular pH, are overexpressed in TME and are involved in adaptative changes occurring in cancer cells to survive at low O2. In this study, we aim to investigate in HNSCC cells and murine models the possibility to target CA IX/XII by the specific inhibitor SLC-0111 to potentiate the effects of cisplatin in hampering cell growth, migration and invasion. Furthermore, we analyzed the signal pathways cooperating in acquisition of a more aggressive phenotype including stemness, epithelial-mesenchymal transition and apoptotic markers. METHODS: The effects of cisplatin, CA IX/XII specific inhibitor SLC-0111, and the combinatorial treatment were tested on proliferation, migration, invasion of HNSCC cells grown in 2D and 3D models. Main signal pathways and the expression of stemness, mesenchymal and apoptotic markers were analyzed by western blotting. Molecular imaging using NIR-Annexin V and NIR-Prosense was performed in HNSCC xenografts to detect tumor growth and metastatic spread. RESULTS: HNSCC cells grown in 2D and 3D models under hypoxic conditions showed increased levels of CA IX/XII and greater resistance to cisplatin than cells grown under normoxic conditions. The addition of CA IX/XII inhibitor SLC-0111 to cisplatin sensitized HNSCC cells to the chemotherapeutic agent and caused a reduction of proliferation, migration and invasiveness. Furthermore, the combination therapy hampered activation of STAT3, AKT, ERK, and EMT program, whereas it induced apoptosis. In HNSCC xenografts the treatment with cisplatin plus SLC-0111 caused an inhibition of tumor growth and an induction of apoptosis as well as a reduction of metastatic spread at a higher extent than single agents. CONCLUSION: Our results highlight the ability of SLC-0111 to sensitize HNSCC to cisplatin by hindering hypoxia-induced signaling network that are shared among mechanisms involved in therapy resistance and metastasis.
Assuntos
Anidrases Carbônicas , Neoplasias de Cabeça e Pescoço , Animais , Anidrases Carbônicas/genética , Anidrases Carbônicas/metabolismo , Anidrases Carbônicas/farmacologia , Proliferação de Células , Cisplatino/farmacologia , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Humanos , Camundongos , Compostos de Fenilureia , Carcinoma de Células Escamosas de Cabeça e Pescoço/tratamento farmacológico , Sulfonamidas , Microambiente TumoralRESUMO
Small extracellular vesicles (sEV) are a class of extracellular vesicles (30-150 nm), delivering molecules including proteins, metabolites, and microRNAs (miRNAs), involved in physiological intercellular crosstalk and disease pathogenesis. The present pilot study aims are (I) to develop an easy and fast protocol for the isolation of sEV from plasma of mast cell tumor (MCT)-affected dogs; (II) to evaluate if miR-21-5p (sEV-miR-21-5p), a miRNA overexpressed by MCT, is associated with sEV. Seventeen dogs have been enrolled in the study: 4 healthy and 13 (6 with and 7 without nodal metastasis) MCT-affected dogs. sEV were isolated using size exclusion chromatography (SEC) (IZON column 35nm) and were characterized by Western blot, Nanoparticle tracking analysis, and transmission electron microscopy. sEV-miR-21-5p was quantified using digital PCR. sEV expressed the specific markers CD9 and TSG101, and a marker of mast cell tryptase. The sEV mean concentration and size were 2.68E + 10 particles/ml, and 99.6 nm, 2.89E + 10 particles/ml and 101.7 nm, and 3.21E + 10 particles/ml and 124 nm in non-metastatic, nodal metastatic, and healthy samples, respectively. The comparative analysis demonstrated that the level of sEV-miR-21-5p was significantly higher in dogs with nodal metastasis compared to healthy (P = 0.038) and without nodal metastasis samples (P = 0.007). In conclusion, the present work demonstrated that a pure population of sEV can be isolated from the plasma of MCT-affected dogs using the SEC approach and that the level of sEV-miR-21-5p is higher in nodal metastatic MCT-affected dogs compared with healthy and MCT-affected dogs without nodal involvement.
RESUMO
BACKGROUND: We have conducted a systematic review focusing on the advancements in preclinical molecular imaging to study the delivery and therapeutic efficacy of miRNAs in mouse models of breast cancer. METHODS: A systematic review of English articles published in peer-reviewed journals using PubMed, EMBASE, BIOSIS™ and Scopus was performed. Search terms included breast cancer, mouse, mice, microRNA(s) and miRNA(s). RESULTS: From a total of 2073 records, our final data extraction was from 114 manuscripts. The most frequently used murine genetic background was Balb/C (46.7%). The most frequently used model was the IV metastatic model (46.8%), which was obtained via intravenous injection (68.9%) in the tail vein. Bioluminescence was the most used frequently used tool (64%), and was used as a surrogate for tumor growth for efficacy treatment or for the evaluation of tumorigenicity in miRNA-transfected cells (29.9%); for tracking, evaluation of engraftment and for response to therapy in metastatic models (50.6%). CONCLUSIONS: This review provides a systematic and focused analysis of all the information available and related to the imaging protocols with which to test miRNA therapy in an in vivo mice model of breast cancer, and has the purpose of providing an important tool to suggest the best preclinical imaging protocol based on available evidence.
RESUMO
BACKGROUND: Management of triple-negative breast cancer (TNBC) is still challenging because of its aggressive clinical behavior and limited targeted treatment options. Cisplatin represents a promising chemotherapeutic compound in neoadjuvant approaches and in the metastatic setting, but its use is limited by scarce bioavailability, severe systemic side effects and drug resistance. Novel site-directed aptamer-based nanotherapeutics have the potential to overcome obstacles of chemotherapy. In this study we investigated the tumor targeting and the anti-tumorigenic effectiveness of novel cisplatin-loaded and aptamer-decorated nanosystems in TNBC. METHODS: Nanotechnological procedures were applied to entrap cisplatin at high efficacy into polymeric nanoparticles (PNPs) that were conjugated on their surface with the epidermal growth factor receptor (EGFR) selective and cell-internalizing CL4 aptamer to improve targeted therapy. Internalization into TNBC MDA-MB-231 and BT-549 cells of aptamer-decorated PNPs, loaded with BODIPY505-515, was monitored by confocal microscopy using EGFR-depleted cells as negative control. Tumor targeting and biodistribution was evaluated by fluorescence reflectance imaging upon intravenously injection of Cyanine7-labeled nanovectors in nude mice bearing subcutaneous MDA-MB-231 tumors. Cytotoxicity of cisplatin-loaded PNPs toward TNBC cells was evaluated by MTT assay and the antitumor effect was assessed by tumor growth experiments in vivo and ex vivo analyses. RESULTS: We demonstrate specific, high and rapid uptake into EGFR-positive TNBC cells of CL4-conjugated fluorescent PNPs which, when loaded with cisplatin, resulted considerably more cytotoxic than the free drug and nanovectors either unconjugated or conjugated with a scrambled aptamer. Importantly, animal studies showed that the CL4-equipped PNPs achieve significantly higher tumor targeting efficiency and enhanced therapeutic effects, without any signs of systemic toxicity, compared with free cisplatin and untargeted PNPs. CONCLUSIONS: Our study proposes novel and safe drug-loaded targeted nanosystems for EGFR-positive TNBC with excellent potential for the application in cancer diagnosis and therapy.
Assuntos
Cisplatino/uso terapêutico , Receptores ErbB/metabolismo , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Animais , Cisplatino/farmacologia , Humanos , Camundongos , Nanopartículas , Técnica de Seleção de AptâmerosRESUMO
BACKGROUND: Invadopodia are actin-based cell-membrane protrusions associated with the extracellular matrix degradation accompanying cancer invasion. The elucidation of the molecular mechanisms leading to invadopodia formation and activity is central for the prevention of tumor spreading and growth. Protein tyrosine kinases such as Src are known to regulate invadopodia assembly, little is however known on the role of protein tyrosine phosphatases in this process. Among these enzymes, we have selected the tyrosine phosphatase Shp1 to investigate its potential role in invadopodia assembly, due to its involvement in cancer development. METHODS: Co-immunoprecipitation and immunofluorescence studies were employed to identify novel substrate/s of Shp1AQ controlling invadopodia activity. The phosphorylation level of cortactin, the Shp1 substrate identified in this study, was assessed by immunoprecipitation, in vitro phosphatase and western blot assays. Short interference RNA and a catalytically-dead mutant of Shp1 expressed in A375MM melanoma cells were used to evaluate the role of the specific Shp1-mediated dephosphorylation of cortactin. The anti-invasive proprieties of glycerophosphoinositol, that directly binds and regulates Shp1, were investigated by extracellular matrix degradation assays and in vivo mouse model of metastasis. RESULTS: The data show that Shp1 was recruited to invadopodia and promoted the dephosphorylation of cortactin at tyrosine 421, leading to an attenuated capacity of melanoma cancer cells to degrade the extracellular matrix. Controls included the use of short interference RNA and catalytically-dead mutant that prevented the dephosphorylation of cortactin and hence the decrease the extracellular matrix degradation by melanoma cells. In addition, the phosphoinositide metabolite glycerophosphoinositol facilitated the localization of Shp1 at invadopodia hence promoting cortactin dephosphorylation. This impaired invadopodia function and tumor dissemination both in vitro and in an in vivo model of melanomas. CONCLUSION: The main finding here reported is that cortactin is a specific substrate of the tyrosine phosphatase Shp1 and that its phosphorylation/dephosphorylation affects invadopodia formation and, as a consequence, the ability of melanoma cells to invade the extracellular matrix. Shp1 can thus be considered as a regulator of melanoma cell invasiveness and a potential target for antimetastatic drugs. Video abstract.
Assuntos
Cortactina/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 6/metabolismo , Pseudópodes/metabolismo , Animais , Linhagem Celular Tumoral , Matriz Extracelular/metabolismo , Humanos , Fosfatos de Inositol/metabolismo , Neoplasias Pulmonares/secundário , Melanoma/metabolismo , Melanoma/patologia , Camundongos Endogâmicos BALB C , Camundongos Nus , Modelos Biológicos , Invasividade Neoplásica , Fosforilação , Ligação Proteica , Especificidade por SubstratoRESUMO
The use of platelet-rich plasma (PRP) to enhance tenodesmic lesion healing has been questioned over the years. The aim of this study was to evaluate current literature to establish the effectiveness of PRP for treating tenodesmic lesions through a systematic review, in accordance with the PRISMA guidelines, and a meta-analysis. Studies comparing PRP with placebo or other treatments for horses with tenodesmic injuries or evaluated PRP effect on tendon and ligament explants were included. Outcomes were clinical, ultrasound, histologic, molecular evaluation, and adverse effects. Two authors independently extracted data and assessed each study's risk of bias. Treatment effects were evaluated using risk ratios for dichotomous data, together with 95% CI. Data were pooled using the random-effects model. The quality of the evidence for each outcome was assessed using GRADE criteria. Twenty-four trials met inclusion criteria for systematic review, while fifteen studies were included in the meta-analysis. Results showed no significant differences in the outcomes between PRP and control groups. Finally, there is no definitive evidence that PRP enhances tendons and ligaments healing. Therefore, there is a need for more controlled trials to draw a firmer conclusion about the efficacy of PRP as a treatment for tenodesmic lesions in the horse.
RESUMO
In a previous study, an ultrasonographic method to assess kidney size in dogs as a ratio of kidney length to aortic luminal diameter (KL/AoD ratio) was proposed. The main limitation of this method was the wide range of normal values (5.5-9.1), which resulted in poor sensitivity and specificity. The aim of this prospective, observational, reference interval study was to determine whether the KL/AoD normal cut-off values in a single breed (Whippets) would have a narrower range than the previously reported normal reference ranges. The influence of sex, age, weight, and side on kidney length (KL) and of sex, age, weight, and scanning plane (longitudinal vs transversal) on aortic luminal diameter (AoD) were also investigated. Thirty-six clinically healthy Whippets (16 males, 20 females) without ultrasonographic renal lesions were included in this study. The 95% confidence interval of mean KL/AoD was found to be narrower than the previously reported range (ie, 6.3-6.9 versus 5.5-9.1). This was considered to be especially notable in that the KL in this breed exhibits marked sexual dimorphism. The KL/AoD ratio did not differ between right versus left sides or male versus female sexes in Whippets (P > .05). Findings from the current study provided KL/AoD ratio normal reference range cut-off values for future use in Whippets and supported the use of breed-specific KL/AoD ratio values for characterizing abnormal renal size in other canine breeds.
Assuntos
Aorta/diagnóstico por imagem , Cães , Rim/diagnóstico por imagem , Ultrassonografia/veterinária , Animais , Feminino , Masculino , Valores de Referência , Sensibilidade e Especificidade , Ultrassonografia/normasRESUMO
Equine exertional rhabdomyolysis (ER) is a well-recognized clinical syndrome affecting racehorses. Prevalence analysis of ER showed that female sex was a significant risk factor. The aim of this research was to evaluate the differences and correlations in the serum activity of muscle enzymes and the stage of the estrous cycle in ER-susceptible and control (C) mares. Serum muscle enzyme activity before and after exercise and sex hormones were analyzed in the two groups of mares. Ten cyclic ER and 10 cyclic C mares were examined weekly for 4 weeks. During diestrus, ER horses had significantly higher resting and postexercise aspartate aminotransferase (AST) activity, but not creatine kinase (CK) activity, compared with controls; only postexercise AST activity was significantly higher during estrus compared with activity levels in controls. During estrus, 17ß-estradiol and AST activity were significantly negatively correlated in the control but not ER mares. Based on our results, further studies should be performed to characterize the presumptive different roles played by sexual hormones in horses susceptible to ER compared with healthy mares.
Assuntos
Doenças dos Cavalos , Condicionamento Físico Animal , Rabdomiólise , Animais , Creatina Quinase , Feminino , Cavalos , Itália/epidemiologia , Músculos , Rabdomiólise/veterináriaRESUMO
The translation of results from the preclinical to the clinical setting is often anything other than straightforward. Indeed, ideas and even very intriguing results obtained at all levels of preclinical research, i.e., in vitro, on animal models, or even in clinical trials, often require much effort to validate, and sometimes, even useful data are lost or are demonstrated to be inapplicable in the clinic. In vivo, small-animal, preclinical imaging uses almost the same technologies in terms of hardware and software settings as for human patients, and hence, might result in a more rapid translation. In this perspective, magnetic resonance imaging might be the most translatable technique, since only in rare cases does it require the use of contrast agents, and when not, sequences developed in the lab can be readily applied to patients, thanks to their non-invasiveness. The wide range of sequences can give much useful information on the anatomy and pathophysiology of oncologic lesions in different body districts. This review aims to underline the versatility of this imaging technique and its various approaches, reporting the latest preclinical studies on thyroid, breast, and prostate cancers, both on small laboratory animals and on human patients, according to our previous and ongoing research lines.
RESUMO
BACKGROUND: Ultrasonography (US) is the recommended imaging technique to evaluate jugular veins. This prospective randomized clinical study was designed to collect a series of B-mode US measurements of manually distended jugular veins in healthy Italian Standardbreds and to find possible correlations between ultrasound measurements and animal morphometric characteristics. Forty-two horses, eight males and 34 females (range 3-22 years; bodyweight 494.4 ± 41.7 kg), were included in the study. The diameters and wall thicknesses of both jugular veins were measured at three different sites of the neck. The differences in ultrasound measurements based on scans, age, gender, side, and site of the neck were evaluated by ANOVA or by the Kruskal-Wallis test. The effects of the morphometric measures on each ultrasound parameter were evaluated by MANOVA (P < 0.05). RESULTS: The ultrasound measurements did not differ significantly between the three different sites or between genders; hence, they were pooled together in the results. On the transverse scan, the mean dorsoventral and lateromedial diameters were 1.58 ± 0.23 and 2.20 ± 0.25 cm, respectively; the mean superficial and deep wall thicknesses (SWT and DWT) were 0.07 ± 0.01 and 0.08 ± 0.01 cm, respectively. On the longitudinal scan, the mean dorsoventral diameter was 1.59 ± 0.26 cm, and the SWT and DWT were both 0.08 ± 0.01 cm. Neck length, from the caudal edge of the mandible to the thoracic inlet, was related to the dorsoventral diameter in both longitudinal and transverse scan and to the SWT and DWT in transverse scan, whereas height at the withers (measured with tape) and estimated weight were related to the wall thickness. Dividing the subjects into groups by age in years ("young" 3-7, "mature" 8-14, "old" > 14), differences were found for the lateromedial diameter in the transverse scan and the SWT on the longitudinal scan. The main limitation of this study was that only one operator performed the measurements. CONCLUSION: The US measurements of the jugular veins and their relationship with morphometric measures reported in this manuscript might be considered as guidelines both for early diagnosis and monitoring jugular vein abnormalities in healthy Italian Standardbred horses.
Assuntos
Cavalos/anatomia & histologia , Veias Jugulares/diagnóstico por imagem , Ultrassonografia/veterinária , Fatores Etários , Animais , Feminino , Veias Jugulares/anatomia & histologia , Masculino , Estudos ProspectivosRESUMO
Junctional adhesion molecule A (JAM-A) is a transmembrane protein that contributes to different biological process, including the epithelial to mesenchymal transition (EMT). Through an EMT profiler array, we explored the molecular players associated with human thyroid cancer progression and identified JAM-A as one of the genes mostly deregulated. The quantitative real-time polymerase chain reaction and immunohistochemistry analyses showed that downregulation of JAM-A occurred in anaplastic thyroid carcinoma (ATC) compared with normal thyroid (NT) and papillary thyroid carcinoma (PTC) tissues and correlated with extrathyroid infiltration, tumor size, and ATC histotype. In ATC cell lines, JAM-A restoration suppressed malignant hallmarks of transformation including cell proliferation, motility, and transendothelial migration. Accordingly, knockdown of JAM-A enhanced thyroid cancer cell proliferation and motility in PTC cells. Through the proteome profiler human phospho-kinase array, we demonstrated that higher expression of JAM-A was associated with a significant increased level of phosphorylation of p53 and GSK3 α/ß proteins. In conclusion, our findings highlight a novel role of JAM-A in thyroid cancer progression and suggest that JAM-A restoration could have potential clinical relevance in thyroid cancer treatment.