Effects of general anaesthesia in dorsal recumbency with and without vatinoxan on bronchoalveolar lavage cytology of healthy horses.

Pneumonia is one of the potential complications of general anaesthesia in horses. Anaesthesia is known to increase neutrophils in bronchoalveolar lavage fluid (BALF) of horses after lateral recumbency, but studies after dorsal recumbency are lacking. Our primary aim was to determine when lung inflammation reaches its maximum and how rapidly BALF cytology returns to baseline after anaesthesia in dorsal recumbency. A secondary aim was to investigate the possible effect of vatinoxan, a novel drug, on the BALF cytology results. Six healthy experimental horses were enrolled in this observational crossover study. The horses were subject to repeated BALF and blood sampling for 7 days after general anaesthesia with two treatment protocols, and without anaesthesia (control). During the two treatments, the horses received either medetomidine-vatinoxan or medetomidine-placebo as premedication, and anaesthesia was induced with ketamine-midazolam and maintained with isoflurane for 1h in dorsal recumbency. The differences in BALF and blood variables between the two anaesthesia protocols and control were analysed with repeated measures analysis of variance models. In this study, anaesthesia in dorsal recumbency resulted in no clinically relevant changes in airway cytology that could be differentiated from the effect of repeated BALF sampling. No differences in BALF matrix metalloproteinase gelatinolytic activity could be detected between the two treatments or the control series. Marked increase in serum amyloid A was detected in some animals. Vatinoxan as premedication did not consistently affect lung cytology or blood inflammatory markers after anaesthesia.

Deficits in distal radius bone strength, density and microstructure are associated with forearm fractures in girls: an HR-pQCT study.

UNLABELLED: Forearm fractures are common during growth. We studied bone strength in youth with a recent forearm fracture. In girls, suboptimal bone strength was associated with fractures. In boys, poor balance and physical inactivity may lead to fractures. Prospective studies will confirm these relationships and identify targets for prevention strategies. INTRODUCTION: The etiology of pediatric forearm fractures is unclear. Thus, we examined distal radius bone strength, microstructure, and density in children and adolescents with a recent low- or moderate-energy forearm fracture and those without forearm fractures. METHODS: We assessed the non-dominant (controls) and non-fractured (cases) distal radius (7% site) using high-resolution peripheral quantitative computed tomography (HR-pQCT) (Scanco Medical AG) in 270 participants (girls: cases n = 47, controls n = 61 and boys: cases n = 88, controls n = 74) aged 8-16 years. We assessed standard anthropometry, maturity, body composition (dual energy X-ray absorptiometry (DXA), Hologic QDR 4500 W) physical activity, and balance. We fit sex-specific logistic regression models for each bone outcome adjusting for maturity, ethnicity, height, and percent body fat. RESULTS: In girls, impaired bone strength (failure load, ultimate stress) and a high load-to-strength ratio were associated with low-energy fractures (odds ratios (OR) 2.8-4.3). Low total bone mineral density (Tt.BMD), bone volume ratio, trabecular thickness, and cortical BMD and thickness were also associated with low-energy fractures (ORs 2.0-7.0). In boys, low Tt.BMD, but not bone strength, was associated with low-energy fractures (OR = 1.8). Boys with low-energy fractures had poor balance and higher percent body fat compared with controls (p < 0.05). Boys with fractures (both types) were less active than controls (p < 0.05). CONCLUSIONS: Forearm fracture etiology appears to be sex-specific. In girls, deficits in bone strength are associated with fractures. In boys, a combination of poor balance, excess body fat, and low physical activity may lead to fractures. Prospective studies are needed to confirm these relationships and clarify targets for prevention strategies.

Discrimination of fractures by low-frequency axial transmission ultrasound in postmenopausal females.

SUMMARY: In this cross-sectional study, 95 postmenopausal women, with and without fracture history, were measured by low-frequency axial transmission ultrasound. The measured ultrasound velocity discriminated the fractured subjects from the nonfractured ones equally or better than peripheral quantitative computed tomography (pQCT) and dual energy x-ray absorptiometry (DXA). These results suggest that low-frequency ultrasound is suitable for bone fragility assessment. INTRODUCTION: Quantitative low-frequency axial transmission ultrasound is a promising modality for assessing mineral density and geometrical properties of long bones such as radius and tibia. The aim of the current study was to evaluate the ability of low-frequency axial transmission ultrasound to discriminate fractures retrospectively in postmenopausal women. METHODS: A cross-sectional study involved 95 female subjects aged 45-88 years, whose fracture information was gathered retrospectively. The fracture group was defined as subjects with one or more low-/moderate-energy fractures. The radius and tibial shaft were measured with a custom-made ultrasonometer to assess the velocity of the low-frequency first-arriving signal (V (LF)). Site-matched pQCT was used to measure volumetric cortical and subcortical bone mineral density (sBMD), and cortical thickness (CTh). Areal BMD (aBMD) was measured using DXA for the whole body (WB), lumbar spine, and hip. RESULTS: The majority (19/32; 59 %) of the fractures were in the upper limb. V (LF) in the radius, but not in the tibia, discriminated fractures with an age- and BMI-adjusted odds ratio (OR) of 2.06 (95 % CI 1.21-3.50, p < 0.01). In the radius, CTh and cortical BMD (CBMD) significantly discriminated fractures, as did the total, cortical, and sBMD in the tibia (adjusted OR 1.35-2.15, p < 0.05). Sensitivity and specificity were similar among all the measurements (area under the receiver operating characteristic curve 0.74-0.81, p < 0.001). CONCLUSIONS: Low-frequency axial transmission ultrasound in the radius was able to discriminate fractured subjects from the nonfractured ones. This suggests that low-frequency axial transmission ultrasound has the potential to assess bone fragility in postmenopausal women.

Matrix metalloproteinases 2 and 9 and their tissue inhibitors in low malignant potential ovarian tumors.

OBJECTIVES: This study aimed to analyze the expression of matrix metalloproteinases 2 (MMP-2) and 9 (MMP-9) and their tissue inhibitors TIMP-1 and TIMP-2 in low malignant potential (LMP) ovarian tumors and to compare these values with those recorded for benign and malignant ovarian neoplasms. METHODS: A total of 53 ovarian tumors (16 benign, 15 LMP and 22 malignant) were evaluated by immunohistochemistry for the expression of MMP-2, MMP-9, TIMP-1 and TIMP-2. RESULTS: MMP-2 expression was found in 56% of the benign, 40% of the LMP, and 90% of the malignant ovarian tumors (benign vs. malignant, p = 0.021; LMP vs. malignant, p = 0.002). The expression of MMP-9, TIMP-1 and TIMP-2 was lower in the benign and LMP tumors compared with the malignant ones. CONCLUSION: The data suggest that, in relation to the expression of MMP-2, MMP-9, TIMP-1 and TIMP-2, LMP ovarian tumors are more similar to benign than to malignant ovarian tumors.

Expression of MMP2, MMP9, MT1-MMP, TIMP1, and TIMP2 mRNA in valvular lesions of the heart.

Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play an important role in several diseases. This study was undertaken to investigate the mRNA synthesis of MMP2, MMP9, membrane-type 1 (MT1)-MMP, and matrix metalloproteinase inhibitors TIMP1 and TIMP2 by in situ hybridization in a set of heart mitral and aortic valves operatively removed due to degenerative or inflammatory valvular diseases. The material consisted of 21 valves, eight with endocarditis and 13 with a degenerative valvular disease. The samples were studied by in situ hybridization with specific probes for MMP2, MMP9, MT1-MMP, TIMP1, and TIMP2. Synthesis of MMP2 mRNA was found in seven valves, five with endocarditis and two with degenerative valvular disease. Signals for MMP9 mRNA were found in two cases with endocarditis and five cases with degenerative valvular disease. No signal for MT1-MMP mRNA was found in the lesions. TIMP1 mRNA, on the other hand, was found in 17 cases, both endocarditis and degenerative valvular disease. TIMP2 mRNA was found in three cases of endocarditis. The signals for MMP2, MMP9, TIMP1, and TIMP2 mRNA were localized in endothelial cells and in fibroblast-like cells expressing alpha-smooth muscle actin, thus showing myofibroblast-type differentiation. The results show that matrix metalloproteinases MMP2 and MMP9, and matrix metalloproteinase inhibitors TIMP1 and TIMP2 mRNAs are synthesized in diseased valves and suggest that they may contribute to matrix remodelling in valvular disease.

Comparative analysis of the distribution of laminin chains in the basement membranes in some malignant epithelial tumors: the alpha1 chain of laminin shows a selected expression pattern in human carcinomas.

Laminins (Ln), together with Type IV collagen and nidogen-1, form the structural integrity of the basement membranes (BM). In this study we used immunohistochemistry to show the distribution of laminin chains alpha1, alpha3, alpha5, beta1, beta2, beta3, gamma1, gamma2, as well as Type IV collagen, in various types of carcinomas and in normal tissues. Except for diffuse gastric carcinomas and infiltrative breast carcinomas, the malignant epithelial tumor clusters were surrounded by quite a continuous BM in most tumors. These BMs comprised most abundantly Ln alpha5, beta1, and gamma1 chains. Conversely, the Ln alpha1 chain, a component of laminins-1 and -3, showed the most restricted distribution in BMs of both normal tissues and malignancies, being moderately present in carcinomas of thyroid gland and ovary and in intraductal carcinomas of breast. In other types of carcinomas, immunoreactivity for Ln alpha1 chain was found more randomly and was practically negative in carcinomas of tongue, stomach, and colon. These findings were comparable to those observed by in situ hybridization, which showed that carcinomas of thyroid gland and intraductal carcinomas of breast constitutively expressed Ln alpha1 mRNA and that the epithelial tumor cells were the main producers of it. The results suggest that epithelial malignancies, except for infiltrative breast and diffuse gastric carcinomas, produce more notable amounts of BM macromolecules in their growth substratum than has previously been anticipated. Corroborating their widespread distribution in normal epithelial tissues, the chains of Lns-5 and -10 are the most abundant Ln molecules in the corresponding carcinomas.

Localization of MT1-MMP, TIMP-1, TIMP-2, and TIMP-3 messenger RNA in normal, hyperplastic, and neoplastic endometrium. Enhanced expression by endometrial adenocarcinomas is associated with low differentiation.

We studied membrane-type matrix metalloproteinase-1 (MT1-MMP), tissue inhibitor of metalloproteinase-1 (TIMP-1), TIMP-2, and TIMP-3 messenger RNA (mRNA) using in situ hybridization to elucidate their temporal and spatial expression patterns in normal, hyperplastic, and neoplastic endometrium. All mRNAs studied were expressed weakly in proliferating endometrium but were induced strongly in late secretory endometrium except MT1-MMP. Endometrial hyperplasia samples did not show increased MT1-MMP or TIMP mRNA expression, indicating that the overall expression patterns in hyperplasia are comparable to those in proliferating endometrium under estrogen effect and that synthesis of extracellular matrix proteins, rather than degradation, predominates in this condition. Exceptionally, stromal cells in areas of desquamation were seen to express focally intense MT1-MMP mRNA in hyperplasia samples. All mRNAs investigated were expressed increasingly in endometrial adenocarcinomas, especially in less differentiated carcinomas. Furthermore, gelatin zymography revealed higher functional degradative activities in carcinoma tissues than in normal endometrium. Our results indicate that MT1-MMP expression, together with that of TIMPs, is involved most notably in normal endometrium under progesterone effect and, without being connected to cyclic hormonal levels, has an important role in the invasive growth of endometrial adenocarcinomas.

Differential expression of matrix metalloproteinase (MMP)-2, MMP-9, and membrane type 1-MMP in hepatocellular and pancreatic adenocarcinoma: implications for tumor progression and clinical prognosis.

In the present study, we used in situ hybridization to study 36 primary hepatocellular carcinomas (HCCs) and 35 pancreatic adenocarcinomas to analyze the expressions of membrane-type 1 matrix metalloproteinase (MT1-MMP), MMP-2, and MMP-9 mRNAs. In HCCs, MT1-MMP mRNA was mainly expressed by cancer cells and to a lesser extent by stromal cells. MMP-2 mRNA was expressed predominantly by cells of tumor stroma, whereas MMP-9 mRNA was seen mainly in neoplastic epithelial cells. In pancreatic adenocarcinomas, MT1-MMP and MMP-9 mRNA were seen at moderate levels both in cancer and in stromal cells, whereas MMP-2 mRNA was predominantly expressed by the tumor stroma. Antigens of MMP-2, MMP-9, and MT1-MMP immunolocalized to the neoplastic epithelium and to the stromal cells in both tumor types. In gelatin zymography, increased amounts of latent and active MMP-2 were found in tumor samples of HCC as compared with adjacent nontumorous liver tissue. On the other hand, the latent form of MMP-9 was found in almost equal amounts both in tumor and normal liver samples, but its active form was present only in HCC. Expression of MT1-MMP mRNA had a tendency to be associated with a lower degree of differentiation in HCC, but such association was not noticed in pancreatic tumors. Correlation to the clinical data showed that MT1-MMP expression had a strong statistical association with a poor outcome of patients (P < 0.01). A similar tendency was also observed in pancreatic adenocarcinomas, but the association did not reach statistical significance. MMP-2 and MMP-9 mRNA expression did not have significant correlation with prognosis. The results of this study support the previous suggestions of the importance of MT1-MMP for malignant growth and indicate that increased MT1-MMP mRNA expression by tumor cells in HCCs and pancreatic adenocarcinomas may have prognostic significance.

Expression of the laminin gamma2 chain in different histological types of lung carcinoma. A study by immunohistochemistry and in situ hybridization.

Sixty-four malignant lung tumours and 12 of their regional lymph node metastases were analysed for expression of the laminin gamma2 chain by immunohistochemistry and in situ hybridization. Expression of the laminin gamma2 chain was strongest in squamous cell carcinomas, followed by adenocarcinomas and large cell carcinomas. Positive cells, except for large cell carcinomas, were located at the epithelial-stromal interface of tumour clusters. An important exception was small cell lung carcinoma, with only a low level of laminin gamma2 chain expression. Apart from tumour type, this may reflect the relatively scanty fibrous stroma in these tumours and supports previous observations that small cell lung carcinoma cells, contrary to other types, lack surface expression of alpha(6)beta(4) integrin, the specific laminin-5 binding receptor. In frozen sections, immunohistochemistry showed linear basement membranes around tumour clusters in squamous cell carcinomas and adenocarcinomas. This shows that carcinoma cells are capable of heavy deposition of the laminin gamma2 chain around tumour clusters and suggests that a laminin gamma2 chain-containing substrate may be of significance for the spread and growth of malignant tumours.

Expression of the laminin gamma 2 chain in pancreatic adenocarcinoma.

Forty-two pancreatic adenocarcinomas were investigated immunohistochemically and by in situ hybridization for the expression of the laminin gamma 2 chain. In 41 cases, intracytoplasmic immunoreactivity for the gamma 2 chain was seen. Positive tumour cells were located especially at the epithelial-stromal interface of the tumour cell islands. In 22 cases, diffuse laminin gamma 2 chain immunoreactivity could also be seen in stroma and in seven cases, occasional positivity was detected in the neoplastic basement membranes. Signals for laminin gamma 2 chain mRNA in tumour cells displayed a distribution similar to that observed on immunohistochemistry. There were significantly more cases with less than 20 per cent of laminin gamma 2 chain-positive tumour cells in tumours extending to peripancreatic tissues and/or tumours with regional or distant metastases (p = 0.029). A corresponding statistical significance could also be noted in the mRNA level (P = 0.025). The results show that pancreatic adenocarcinomas display a high activity of laminin gamma 2 chain synthesis. Tumours with a strong laminin gamma 2 chain synthesis show a lower invasive and metastatic potential than tumours with a weak or moderate laminin gamma 2 chain expression.