Bones are organs of the skeletal system, providing shape, mechanical support, and protection to the body and facilitating the movement. In addition, bones contribute to the mineral homeostasis of the body and have rec...Bones are organs of the skeletal system, providing shape, mechanical support, and protection to the body and facilitating the movement. In addition, bones contribute to the mineral homeostasis of the body and have recently been found to participate in endocrine regulation of energy metabolism. The well-known limitations associated with clinical use of autografts and allografts continue to drive efforts to develop bone graft substitutes, using the principles of biomaterials and tissue engineering. Under some stressful and continuous compressive conditions, the ability of the bone tissue to tolerate strength decreases. Whenever these forces overcome the toleration of the bone tissue, bone fracture occurs. years展开更多
Porcine reproductive and respiratory syndrome(PRRS) is one of the most important diseases of swine industry. The causal agent, PRRS-virus(PRRSV), is able to evade the host immune response and survive in the organism c...Porcine reproductive and respiratory syndrome(PRRS) is one of the most important diseases of swine industry. The causal agent, PRRS-virus(PRRSV), is able to evade the host immune response and survive in the organism causing transient infections. Despite all scientific efforts, there are still some gaps in the knowledge of the pathogenesis of this disease. Antigen presenting cells(APCs), as initiators of the immune response, are located in the first line of defense against microorganisms, and are responsible for antigen recognition, processing and presentation. Dendritic cells(DCs) are the main type of APC involved in antigen presentation and they are susceptible to PRRSV infection. Thus, PRRSV replication in DCs may trigger off different mechanisms to impair the onset of a host effective immune response against the virus. On the one side, PRRSV may impair the basic functions of DCs by regulating the expression of major histocompatibility complex class Ⅱ and CD80/86. Other strategy followed by the virus is the induction of cell death of APCs by apoptosis, necrosis or both of them. The impairment and/or cell death ofAPCs could lead to a failure in the onset of an efficient immune response, as long as cells could not properly activate T cells. Future aspects to take into account are also discussed in this review.展开更多
Double balloon enteroscopy(DBE)is an endoscopic technique broadly used to diagnose and treat small bowel diseases.Among the associated complications of the oral DBE,post-procedure pancreatitis has taken the most atten...Double balloon enteroscopy(DBE)is an endoscopic technique broadly used to diagnose and treat small bowel diseases.Among the associated complications of the oral DBE,post-procedure pancreatitis has taken the most attention due to its gravity and the thought that it might be associated to the technique itself and anatomical features of the pancreas.However,as the etiology has not been clarified yet,this paper aims to review the published literature and adds new results from a porcine animal model.Biochemical markers,histological sections and the vascular perfusion of the pancreas were monitored in the pig during DBE practice.A reduced perfusion of the pancreas and bowel,the presence of defined hypoxic areas and disseminated necrotic zones were found in the pancreatic tissue of pigs.All these evidences contribute to support a vascular distress as the most likely etiology of the post-DBE pancreatitis.展开更多
AIM： To evaluate the effect of double-balloon enteros- copy （DBE） on pancreas histology and levels of pancre- atic enzymes. METHODS： Conventional upper gastrointestinal endos- copy was performed on five control pi...AIM： To evaluate the effect of double-balloon enteros- copy （DBE） on pancreas histology and levels of pancre- atic enzymes. METHODS： Conventional upper gastrointestinal endos- copy was performed on five control pigs. Oral DBE was performed with an EN-450T5 enteroscope on 20 pigs. Two experimental groups （10 pigs each） were defined according to DBE duration： 90 rain for Group 1 and 140 min for Group 2. During oral insertion, the balloons were not inflated in the descending part of the duodenum to avoid the minor duodenal papilla. Serum amy- lase, lipase and C-reactive protein （CRP） levels were monitored before the procedure and repeated every 30 min until the exploration was finished, as well as 24 h and 7 d after. After the procedure and for a total of 7 d, the pigs were observed twice a day for signs of de- creased activity, irritability, vomiting or anorexia. Gross and microscopic examination of the pancreas was per- formed on day 7. RESULTS： All animals tolerated DBE without clini- cal manifestations of acute pancreatitis. Experimental groups had higher levels of enzymes than the control group at 24 h. Throughout the exploration, the amylase levels increased significantly above the baseline 24 h after DBE, although the increase was not statistically significant and did not reach 20% of the baseline. An increase in lipase and CRP was observed at 24 h after the procedure, although by day 7, all enzymatic lev- els had returned to baseline. No differences between Groups 1 and 2 were found for any enzyme and sam- pling site during and after the procedure. Similarly, no correlation between insertion depth and enzyme levels was observed. Direct in situ and post-removal inspec- tion of the pancreas did not show any evidence of fluid collection, abscesses or hemorrhage. Histological examination of the pancreas from Groups 1 and 2 re- vealed the existence of focal areas （0.14-0.26 mm2） of ischemic necrosis in 47.4% of the animals. In the pigs with damaged pancreas, the left lobe （tail） was always affected. However, this only happened in 83.3% of the samples from the right lobe （head） and in 33.3% of the samples from the body of the pancreas. Significant differences were found between the left lobe （tail） and the body for the percentage of affected pancreas. Both the size of the lesions and the percentage of affected pancreas were higher in the left pancreatic lobe （tail）. The presence of the lesions was not related to the ex- ploration length.CONCLUSION： The increase in pancreatic enzymes after DBE could be related to focal points of pancreatic ischemic necrosis due to mechanical stress.展开更多
Epigenetic regulators have been implicated in tumorigenesis of many types of cancer;however,their roles in endothelial cell cancers such as canine hemangiosarcoma(HSA)have not been studied.In this study,we find that l...Epigenetic regulators have been implicated in tumorigenesis of many types of cancer;however,their roles in endothelial cell cancers such as canine hemangiosarcoma(HSA)have not been studied.In this study,we find that lysine-specific demethylase 2 b(KDM2 B)is highly expressed in HSA cell lines compared with normal canine endothelial cells.Silencing of KDM2 B in HSA cells results in increased cell death in vitro compared with the scramble control by inducing apoptosis through the inactivation of the DNA repair pathways and accumulation of DNA damage.Similarly,doxycycline-induced KDM2 B silencing in tumor xenografts results in decreased tumor sizes compared with the control.Furthermore,KDM2 B is also highly expressed in clinical cases of HSA.We hypothesize that pharmacological KDM2 B inhibition can also induce HSA cell death and can be used as an alternative treatment for HSA.We treat HSA cells with GSK-J4,a histone demethylase inhibitor,and find that GSK-J4 treatment also induces apoptosis and cell death.In addition,GSK-J4 treatment decreases tumor size.Therefore,we demonstrate that KDM2 B acts as an oncogene in HSA by enhancing the DNA damage response.Moreover,we show that histone demethylase inhibitor GSK-J4 can be used as a therapeutic alternative to doxorubicin for HSA treatment.展开更多
Nowadays the system of public health is constructed in such a manner so that its main objective is the recovery of an already sick person, while prediction and prevention receive little attention. Meanwhile the develo...Nowadays the system of public health is constructed in such a manner so that its main objective is the recovery of an already sick person, while prediction and prevention receive little attention. Meanwhile the development of these aspects of medicine can lead to the ability to control morbidity among the population, to identify chronic and genetic diseases in the early stages of development, and thus to prevent their further progression. This will reduce traditionally high costs of sick people treatment and the number of disabled population, and improve the quality and duration of life. The elaboration of new fields of science that are working on the study and interpretation of data obtained during laboratory and clinical research, creation of new methods for diagnosis, prognosis and treatment, provides an opportunity now to implement a new strategy, called PPPM, and gets promising results, which should lead to further development of an existing medicine.展开更多
AIM: To determine the hypothesis that inflating the balloons in the duodenal papilla determines changes in the biochemical markers of pancreatitis.METHODS: Four groups of pigs were used: Group papilla(GP), the overtub...AIM: To determine the hypothesis that inflating the balloons in the duodenal papilla determines changes in the biochemical markers of pancreatitis.METHODS: Four groups of pigs were used: Group papilla(GP), the overtube's balloon was inflated in the area of the papilla; GP + double balloon enteroscopy(GP + DBE), the overtube's balloon was kept inflated in the area of the papilla for 20 min before a DBE; Group DBE(GDBE), DBE was carried out after insuring the balloon's inflation far from the pancreatic papilla; and Group control(GC). Serum concentrations of amylase, lipase and C-reactive protein(CRP) were evaluated. Pancreases were processed for histopathology examination.RESULTS: Main changes occurred 24 h after the procedure compared with baseline levels. Amylase levels increased significantly in GP(59.2% higher) and were moderately higher in groups GP + DBE and GDBE(22.7% and 20%, respectively). Lipase increased in GP and GP + DBE, whereas it hardly changed in GDBE and in GC. CRP increased significantly in GP, GP + DBE and GDBE, while no changes were reported for GC. No statistically significant difference between groups GP and GP + DBE was found for the histopathological findings, except for vacuolization and necrosis of the pancreatic parenchyma that was higher in GP than in GP + DBE.CONCLUSION: The manipulation of the duodenal papilla by the inflated overtube's balloon during DBE causes pancreatic structural damage and increased biochemical markers associated with pancreatitis.展开更多
ROSE is a method for fast sample evaluation that does not compromise sensitivity and specificity in the hospital environment when establishing the diagnosis and the therapeutic protocol. The technique is already well-...ROSE is a method for fast sample evaluation that does not compromise sensitivity and specificity in the hospital environment when establishing the diagnosis and the therapeutic protocol. The technique is already well-established in human medicine as it operates within the standard operational procedure, reducing the rates of inconclusive results and cancelled cytopathology assays in the hospital environment. However, its application is scarce in Veterinary Medicine, including in hospitals where intensive care is available. This study aims at conducting a case-by-case assessment of inconclusive and cancelled examinations in the Cytopathology Outpatient Clinic at UNESP (HV), Botucatu, Brazil, from 2012 to 2016 and ascertains the causes. For this purpose, a retrospective study was conducted for 9587 examinations in canines, of which 4.1% and 10.44% were inconclusive and cancelled, respectively. These results are not in line with ROSE, which foresees a total rate of 5% for these occurrences. The reasons for these high rates in the outpatient clinic were the lack of a Standard Operating Procedure (SOP) and the relative inexperience of the residents in a university hospital. Therefore, with the adoption of the ROSE protocol for veterinary cytopathological examinations, together with adequate training for the outpatient professionals responsible for the examinations, the goals set forth by the ROSE protocol would be met and the rates of inconclusive and cancelled examinations would decrease considerably. Lastly, this study hopes to contribute towards the diagnosis and therapeutic protocols of the main diseases affecting dogs.展开更多
Milk production in Ecuador has enormous economic importance and large-, medium- and small-scale producers all participate in the market. There are multiple climatic regions, and dairy production is present in every on...Milk production in Ecuador has enormous economic importance and large-, medium- and small-scale producers all participate in the market. There are multiple climatic regions, and dairy production is present in every one of them. High ambient temperatures in the Ecuadorian tropics represent a key challenge to the conservation of milk in the custody of smallholders. The objective of the present study was to evaluate the efficiency of the application of a chemical activator of the Lactoperoxidase System (LP-s) in the conservation of raw milk, at room temperature, in the Ecuadorian tropics. In the present study, sodium thiocyanate—0.36 g·L-1 of milk—and sodium percarbonate—1.36 g·L-1 of milk—as an activator of LP-s were used and the pH and microbiological characteristics (total coliforms, Staphylococcus aureus, total aerobes, molds and yeasts) of the milk at different storage times (0, 4 and 8 hours). The results obtained in the present study showed a significant difference between the two groups under study at 8 hours of storage at room temperature in all parameters (except yeasts where there was no growth in the two treatments), being relevant the significant decrease of the bacterial content. Thus the present study shows that the use of sodium thiocyanate and sodium percarbonate in the above described concentrations could be modulating the activation of LP-s that provides an efficient alternative for the conservation of the raw milk without refrigeration, improving the income for losses of the product and obtaining a raw material of good quality for sale or for further processing, mainly for small producers who do not have the economic resources to have refrigeration means for their product and who must transport their milk for considerably longer distances until they arrive at the collection centers or the processing plants for sale, thus showing that the method used in the present study is not only effective but also has a relatively low cost and easy application.展开更多
Milk is one of the products that can be adulterated in many ways affecting the quality of this and its derivatives. Glucomacropeptide (GMP) is a protein that is found only in the whey from the production of fresh chee...Milk is one of the products that can be adulterated in many ways affecting the quality of this and its derivatives. Glucomacropeptide (GMP) is a protein that is found only in the whey from the production of fresh cheese, enzymatically obtained from the coagulation of casein and which is commonly used to adulterate fresh or powdered milk. The aim of this study was to determine the adulteration of milk with cheese whey thought a molecular approach, where the glucomacropeptide was collected by sequential precipitation with trichloroacetic acid (ATC) and detected by polyacrylamidododecylsulfate gel electrophoresis (PAGE-SDS), using samples of fresh milk, intentionally adulterated with serum in the proportion of 0%, 1%, 5%, 10% and 15%. The results obtained showed that the detection of glucomacropeptide by electrophoresis was positive in all samples of adulterated milk, evidencing a band of 20.9 kDa in the reading, corresponding to the molecular weight of the GMP, showing that the technique used determines the adulteration in the milk, in a specific and sensitive way, also shows that in the evaluation of physical-chemical and microbiological parameters of milk, there are no significant differences between treatments, except for the pH that tends to decrease as the percentage of serum in the milk increases.展开更多
The sarcoma is the generic nomenclature for neoplasm of mesodermal cells, which express in man and animals. Silent growth requires early diagnosis technique for identifying their proteins. The experimental model in vi...The sarcoma is the generic nomenclature for neoplasm of mesodermal cells, which express in man and animals. Silent growth requires early diagnosis technique for identifying their proteins. The experimental model in vivo murine sarcoma 180-TG (TG-180), is widely used in research to provide the stimuli of infectious and neoplastic antigens. In this case, the technique of immunohisto-chemistry helps identify the expressions of tumor cell variants. The objective of the research was to characterize immunoexpression murine sarcoma TG 180, by immunohistochemistry, antibodies AE1/AE3, vimentin, CD3, CD 45, CD79α and S100A4. For this, murine sarcoma TG-180, was implanted subcutaneously in 20 mice “Swiss”, male, 30 days old, 28 g for 10 days. Samples were taken and subjected to immunohistochemistry, via use of HistoMouse-MA™? kit. There was specifically labeled S100A4 and vimentin antibodies, indicative of poorly differentiated neoplasms fibroblasts. In fact, the model is established by identifying the origin of the cell, once identified, chemotherapeutic tests can also be performed. Neoplasia like these, when installed in man and animals, depending on the degree of aggressiveness requires treatment protocol varied between surgery and chemotherapy or combination of treatments.展开更多
Background: Several animal models, including dogs, have been useful to compare the pathogenesis of mammary neoplasm in humans, showing biological parallelism in the growth and development of breast cancer. The causes ...Background: Several animal models, including dogs, have been useful to compare the pathogenesis of mammary neoplasm in humans, showing biological parallelism in the growth and development of breast cancer. The causes of cancer could be attributed to change in several tumor suppressor genes. The relationship between molecule associated to senescence and clinical prognosis of patients affected by mammary cancer is little known. Beyond a collection of data, the major interest of the present study was to carry out a clinical follow-up of patients affected by these tumors, through association with new molecular markers by immunohistochemical technic. Upon completing the study, 15 patients survived, while 45 died. In the case of malignant neoplasms, 40 patients died because of the illness. The type of surgery most used by veterinarian surgeons was the simple lumpectomy, followed by the regional mastectomy. Sentinel node was removed by surgery only when clearly affected. Result: Markings against steroid hormones were positive. Regarding the markings against HER2 and Ki-67, they were negative in all cases. The markings against P53 and CD31 were all positives. Markings against molecules associated with cellular senescence were all positives. No statistical differences were found in immunomarcation for the different antigens used as clinical prognosis factors in mammary neoplasms. Conclusions: According to the study conditions, the survival of patients affected by breast tumors is directly related to diagnosis and malignancy histological grade, but not to animal breed, number of affected glands or patient reproductive status. On the other hand, immunohistochemical markings were not related to the patient prognosis. For this reason, it is important to highlight the persistance of a high percentage of mammary neoplasm cases clinically diagnosed with poor results on patient survival. Thus, educating owners and veterinarians for using diagnostic available tools to improve the prognosis after surgical animals affected by breast cancer is quite necessary.展开更多
Background: Experimental studies have shown that cyclo-oxygenase-2 (Cox2) is related to the development and progression of tumors, since this enzyme is induced and expressed by cells such as macrophages, osteoblasts, ...Background: Experimental studies have shown that cyclo-oxygenase-2 (Cox2) is related to the development and progression of tumors, since this enzyme is induced and expressed by cells such as macrophages, osteoblasts, “activated” endothelial cells, and tumor cells. The activity in tumors includes proliferation, cell transformation, tumor growth, invasion and metastasis and may play an important role in carcinogenesis of the canine osteosarcoma, since it has high expression in tissue fragments. The combination of selective Cox2 inhibitors and other treatment modalities is the basis for a new anti-cancer therapy strategy. This in vitro study exposed primary cells of five different canine osteosarcoma cultures to selective Cox2 inhibitor at increasing concentrations and times. Results: For Cox2 negative cultures, despite the absence of differences, greater sensitivity of cells to treatment was observed. For Cox2 positive cultures, a higher number of necrotic cells were observed (P ≤ 0.05), when compared with negative cultures. For exposure times with Celecoxib doses, no difference (P > 0.05) was found between the three times analyzed for living, apoptotic and apoptotic/necrotic cells. There are similarities in the values of 24 h and 48 h, with slight reduction of living cells, increasing those undergoing apoptosis and apoptosis/necrosis. There was significance for necrosis (P ≤ 0.05). In 72 hours, a significant difference was observed between the other two previous values (P ≤ 0.05). It was found for the group of 100 μML-1, that there was a numerically greater signaling for apoptosis and lower (P = 0.08) for necrosis, and this point was the onset of the pharmacodynamic phenomenon, with drop in the values for living cells and increased number of necrotic cells, with a tendency (P = 0.08) for reducing the percentage of necrotic cells for the group of 100 μML-1 when compared to that of 10 μML-1. Conclusions: For Cox2 positive and negative cultures, there was difference for necrotic cells and there was no difference between Cox2 positive and Cox2 negative groups in relation to the percentage of living cells and apoptotic and apoptotic/necrotic cells. At time of 72 hours, higher percentage of living cells, lower percentage of apoptotic cells and increased percentage of necrotic cells in relation to groups of 24 and 48 hours was observed. A tendency for reducing the percentage of necrotic cells for the group of 100 μML-1 when compared to that of the group of 10 μML-1 was observed.展开更多
Diagnosis and biological behavior of breast cancer of female dog represent one of the biggest challenges facing the Veterinarian in recent years. Due to its exponential growth and the degree of aggressiveness, the exa...Diagnosis and biological behavior of breast cancer of female dog represent one of the biggest challenges facing the Veterinarian in recent years. Due to its exponential growth and the degree of aggressiveness, the exact cause of this tumor is probably multifactorial and it is believed that may suffer influence from environmental factors. Among the suspected environmental contaminants are the pyrethroids. Aiming to investigate the participation of pyrethroids in tumorigenesis in female dogs, a study was conducted using 50 female dogs, 22 were positive for simple breast carcinoma (Group I), 18 with a diagnosis of complex breast carcinoma (Group II) and 10 negative (Group III) for breast cancer. In order to detect DNA damage, the Comet assay was performed on mammary samples of these animals, which also had samples submitted to the technique of High Performance Liquid Chromatography (HPLC), which aimed to quantify the concentration of pyrethroids. The results of HPLC of each animal were compared with those obtained by the Comet assay analysis of variance and the means were compared by the test groups “Student T” at the significance level of p ￡ 0.05. Despite presenting correlation between the amount of DNA damage and tumor aggressiveness, no statistical differences were found in the DNA damage of different histologic types of breast carcinoma. As for pyrethroids, even these were detected in 22% of tumor tissues and peritumoral fat, there was no difference in DNA damage between cells exposed and not exposed to environmental contaminant.展开更多
Objective: To investigate the potential of superparamagnetic iron oxide particles (SPIO) in MR imaging for the differentiation between hyperplastic and metastatic lymph node. Methods: Animal models of malignant lymph ...Objective: To investigate the potential of superparamagnetic iron oxide particles (SPIO) in MR imaging for the differentiation between hyperplastic and metastatic lymph node. Methods: Animal models of malignant lymph node metastasis were established in 6 New-Zealand rabbits by a unilateral intra-muscular injection of VX2 carcinoma cells, and models of hyperplastic lymph nodes were induced in another 6 rabbits by a unilateral intra-muscular injection of egg yolk emulsion. MR images of the lymph nodes were obtained before and 12 h after interstitial injection of SPIO. Image results were analyzed and compared with pathological findings. Results: On unenhanced images, the signal intensity of hyperplastic and metastatic lymph nodes did not differ significantly. After administration of SPIO, the signal intensity of both hyperplastic and metastatic lymph nodes remained unchanged on T1-weighted SE images. On T2-weighted SE images, the signal intensity of hyperplastic lymph nodes decreased heterogeneously, while that of all metastatic ones remained unchanged. On T2-weighted GRE images, the signal intensity of hyperplastic lymph nodes decreased significantly and homogeneously, while that of 4 metastatic ones remained unchanged and that of the rest 2 decreased heterogeneously. Conclusion: SPIO-enhanced MR imaging may enable the differentiation between the hyperplastic and metastatic lymph nodes.展开更多
Intestinal stem cells(ISCs)promote tissue repair after genotoxic or immune-mediated injury.However,ISCs are particularly sensitive to various stressors and primary targets of overwhelming immune responses,such as inte...Intestinal stem cells(ISCs)promote tissue repair after genotoxic or immune-mediated injury.However,ISCs are particularly sensitive to various stressors and primary targets of overwhelming immune responses,such as interferonγ(IFNγ)-mediated killing.In mouse models of radiation therapy-induced gut damage and in biopsies from patients who underwent allogeneic hematopoietic stem cell transplantation,we observed IFNγexpression by intestinal Treg cells.Treg cells leverage combined IFNγand interleukin 10(IL-10)stimulation of ISCs to nurture the growth of intestinal organoids through the activation of the mTORC1 and Myc pathways.Similarly,Treg cells or the combined addition of recombinant IFNγand IL-10 promoted the regeneration of organoids after irradiation,and both cytokines were essential for ensuring epithelial regeneration following acute intestinal tissue injury in vivo.The exposure of organoids to growth factor-free culture conditions revealed distinct EGF-like properties of IFNγand Wnt-like properties of IL-10.While IFNγrapidly induced epithelial proliferation,it depleted the pool of ISCs in vitro.Only the combination of IFNγand IL10 led to epithelial proliferation and organoid growth while simultaneously ensuring ISC maintenance over time.Our results reveal a context-dependent role of inflammatory signaling in ISCs,through which Treg cells promote epithelial repair following therapyinduced injury.展开更多
Halogenated natural products(HNPs)are considered to be emerging contaminants whose environmental distribution and fate are only incompletely known.Therefore,several persistent and bioaccumulative HNP groups,together w...Halogenated natural products(HNPs)are considered to be emerging contaminants whose environmental distribution and fate are only incompletely known.Therefore,several persistent and bioaccumulative HNP groups,together with manmade polychlorinated biphenyls(PCBs)and polybrominated diphenyl ethers(PBDEs),were quantified in the blubber of nine sperm whales(Physeter macrocephalus)stranded on the coast of the Mediterranean Sea in Italy.The naturally occurring polybrominated hexahydroxanthene derivatives(PBHDs;sum of TetraBHD and TriBHD)were the most prominent substance class with up to 77,000 ng/g blubber.The mean PBHD content(35,800 ng/g blubber)even exceeded the one of PCBs(28,400 ng/g blubber),although the region is known to be highly contaminated with manmade contaminants.Based on mean values,Q1∼PBDEs>MeO-BDEs∼2,2′-diMeO-BB 80 and several other HNPs followed with decreasing amounts.All blubber samples contained an abundant compound whose molecular formula(C_(16)H_(19)Br_(3)O_(2))was verified using high-resolution mass spectrometry.The only plausible matching isomer was(2S,4′S,9R,9′S)-2,7-dibromo-4′-bromomethyl-1,1-dimethyl-2,3,4,4′,9,9′-9,9′-hexahydro-1H-xanthen-9-ol(OH-TriBHD),a hydroxylated secondary metabolite previously detected together with TriBHD and TetraBHD in a sponge known to be a natural producer of PBHDs.The estimated mean amount of the presumed OH-TriBHD was 3000 ng/g blubber,which is unexpectedly high for hydroxylated compounds in the lipids of marine mammals.展开更多
文摘Bones are organs of the skeletal system, providing shape, mechanical support, and protection to the body and facilitating the movement. In addition, bones contribute to the mineral homeostasis of the body and have recently been found to participate in endocrine regulation of energy metabolism. The well-known limitations associated with clinical use of autografts and allografts continue to drive efforts to develop bone graft substitutes, using the principles of biomaterials and tissue engineering. Under some stressful and continuous compressive conditions, the ability of the bone tissue to tolerate strength decreases. Whenever these forces overcome the toleration of the bone tissue, bone fracture occurs. years
基金Supported by The Spanish Ministry of Education and Science,No.AGL2009-12438/GAN
文摘Porcine reproductive and respiratory syndrome(PRRS) is one of the most important diseases of swine industry. The causal agent, PRRS-virus(PRRSV), is able to evade the host immune response and survive in the organism causing transient infections. Despite all scientific efforts, there are still some gaps in the knowledge of the pathogenesis of this disease. Antigen presenting cells(APCs), as initiators of the immune response, are located in the first line of defense against microorganisms, and are responsible for antigen recognition, processing and presentation. Dendritic cells(DCs) are the main type of APC involved in antigen presentation and they are susceptible to PRRSV infection. Thus, PRRSV replication in DCs may trigger off different mechanisms to impair the onset of a host effective immune response against the virus. On the one side, PRRSV may impair the basic functions of DCs by regulating the expression of major histocompatibility complex class Ⅱ and CD80/86. Other strategy followed by the virus is the induction of cell death of APCs by apoptosis, necrosis or both of them. The impairment and/or cell death ofAPCs could lead to a failure in the onset of an efficient immune response, as long as cells could not properly activate T cells. Future aspects to take into account are also discussed in this review.
基金Supported by Fundacion Seneca,Comunidad Autonoma de la Region de Murcia,Spain,No.12024/PI/09
文摘Double balloon enteroscopy(DBE)is an endoscopic technique broadly used to diagnose and treat small bowel diseases.Among the associated complications of the oral DBE,post-procedure pancreatitis has taken the most attention due to its gravity and the thought that it might be associated to the technique itself and anatomical features of the pancreas.However,as the etiology has not been clarified yet,this paper aims to review the published literature and adds new results from a porcine animal model.Biochemical markers,histological sections and the vascular perfusion of the pancreas were monitored in the pig during DBE practice.A reduced perfusion of the pancreas and bowel,the presence of defined hypoxic areas and disseminated necrotic zones were found in the pancreatic tissue of pigs.All these evidences contribute to support a vascular distress as the most likely etiology of the post-DBE pancreatitis.
基金Supported by PI070712 Project (Instituto de Salud Carlos Ⅲ, Ministerio de Sandiady Consumo, Spain)BIO-MED07/08-0019 Project (Consejería de Educación, Cienciae Investigación de la Región de Murcia, Spain)
文摘AIM： To evaluate the effect of double-balloon enteros- copy （DBE） on pancreas histology and levels of pancre- atic enzymes. METHODS： Conventional upper gastrointestinal endos- copy was performed on five control pigs. Oral DBE was performed with an EN-450T5 enteroscope on 20 pigs. Two experimental groups （10 pigs each） were defined according to DBE duration： 90 rain for Group 1 and 140 min for Group 2. During oral insertion, the balloons were not inflated in the descending part of the duodenum to avoid the minor duodenal papilla. Serum amy- lase, lipase and C-reactive protein （CRP） levels were monitored before the procedure and repeated every 30 min until the exploration was finished, as well as 24 h and 7 d after. After the procedure and for a total of 7 d, the pigs were observed twice a day for signs of de- creased activity, irritability, vomiting or anorexia. Gross and microscopic examination of the pancreas was per- formed on day 7. RESULTS： All animals tolerated DBE without clini- cal manifestations of acute pancreatitis. Experimental groups had higher levels of enzymes than the control group at 24 h. Throughout the exploration, the amylase levels increased significantly above the baseline 24 h after DBE, although the increase was not statistically significant and did not reach 20% of the baseline. An increase in lipase and CRP was observed at 24 h after the procedure, although by day 7, all enzymatic lev- els had returned to baseline. No differences between Groups 1 and 2 were found for any enzyme and sam- pling site during and after the procedure. Similarly, no correlation between insertion depth and enzyme levels was observed. Direct in situ and post-removal inspec- tion of the pancreas did not show any evidence of fluid collection, abscesses or hemorrhage. Histological examination of the pancreas from Groups 1 and 2 re- vealed the existence of focal areas （0.14-0.26 mm2） of ischemic necrosis in 47.4% of the animals. In the pigs with damaged pancreas, the left lobe （tail） was always affected. However, this only happened in 83.3% of the samples from the right lobe （head） and in 33.3% of the samples from the body of the pancreas. Significant differences were found between the left lobe （tail） and the body for the percentage of affected pancreas. Both the size of the lesions and the percentage of affected pancreas were higher in the left pancreatic lobe （tail）. The presence of the lesions was not related to the ex- ploration length.CONCLUSION： The increase in pancreatic enzymes after DBE could be related to focal points of pancreatic ischemic necrosis due to mechanical stress.
基金supported by the Sasakawa Scientific Research Grant(KG,Research No.2019-4111)provided by the Japan Science Society and the KAKENHI Grant-in-Aid for Young Scientist(KA,Number 18K14575,20K15654)provided by Japan Society for the Promotion of Science。
文摘Epigenetic regulators have been implicated in tumorigenesis of many types of cancer;however,their roles in endothelial cell cancers such as canine hemangiosarcoma(HSA)have not been studied.In this study,we find that lysine-specific demethylase 2 b(KDM2 B)is highly expressed in HSA cell lines compared with normal canine endothelial cells.Silencing of KDM2 B in HSA cells results in increased cell death in vitro compared with the scramble control by inducing apoptosis through the inactivation of the DNA repair pathways and accumulation of DNA damage.Similarly,doxycycline-induced KDM2 B silencing in tumor xenografts results in decreased tumor sizes compared with the control.Furthermore,KDM2 B is also highly expressed in clinical cases of HSA.We hypothesize that pharmacological KDM2 B inhibition can also induce HSA cell death and can be used as an alternative treatment for HSA.We treat HSA cells with GSK-J4,a histone demethylase inhibitor,and find that GSK-J4 treatment also induces apoptosis and cell death.In addition,GSK-J4 treatment decreases tumor size.Therefore,we demonstrate that KDM2 B acts as an oncogene in HSA by enhancing the DNA damage response.Moreover,we show that histone demethylase inhibitor GSK-J4 can be used as a therapeutic alternative to doxorubicin for HSA treatment.
文摘Nowadays the system of public health is constructed in such a manner so that its main objective is the recovery of an already sick person, while prediction and prevention receive little attention. Meanwhile the development of these aspects of medicine can lead to the ability to control morbidity among the population, to identify chronic and genetic diseases in the early stages of development, and thus to prevent their further progression. This will reduce traditionally high costs of sick people treatment and the number of disabled population, and improve the quality and duration of life. The elaboration of new fields of science that are working on the study and interpretation of data obtained during laboratory and clinical research, creation of new methods for diagnosis, prognosis and treatment, provides an opportunity now to implement a new strategy, called PPPM, and gets promising results, which should lead to further development of an existing medicine.
基金Supported by the Project 12024/PI/09(Fundacion SenecaComunidad Autonoma de la Region de MurciaSpain)
文摘AIM: To determine the hypothesis that inflating the balloons in the duodenal papilla determines changes in the biochemical markers of pancreatitis.METHODS: Four groups of pigs were used: Group papilla(GP), the overtube's balloon was inflated in the area of the papilla; GP + double balloon enteroscopy(GP + DBE), the overtube's balloon was kept inflated in the area of the papilla for 20 min before a DBE; Group DBE(GDBE), DBE was carried out after insuring the balloon's inflation far from the pancreatic papilla; and Group control(GC). Serum concentrations of amylase, lipase and C-reactive protein(CRP) were evaluated. Pancreases were processed for histopathology examination.RESULTS: Main changes occurred 24 h after the procedure compared with baseline levels. Amylase levels increased significantly in GP(59.2% higher) and were moderately higher in groups GP + DBE and GDBE(22.7% and 20%, respectively). Lipase increased in GP and GP + DBE, whereas it hardly changed in GDBE and in GC. CRP increased significantly in GP, GP + DBE and GDBE, while no changes were reported for GC. No statistically significant difference between groups GP and GP + DBE was found for the histopathological findings, except for vacuolization and necrosis of the pancreatic parenchyma that was higher in GP than in GP + DBE.CONCLUSION: The manipulation of the duodenal papilla by the inflated overtube's balloon during DBE causes pancreatic structural damage and increased biochemical markers associated with pancreatitis.
文摘ROSE is a method for fast sample evaluation that does not compromise sensitivity and specificity in the hospital environment when establishing the diagnosis and the therapeutic protocol. The technique is already well-established in human medicine as it operates within the standard operational procedure, reducing the rates of inconclusive results and cancelled cytopathology assays in the hospital environment. However, its application is scarce in Veterinary Medicine, including in hospitals where intensive care is available. This study aims at conducting a case-by-case assessment of inconclusive and cancelled examinations in the Cytopathology Outpatient Clinic at UNESP (HV), Botucatu, Brazil, from 2012 to 2016 and ascertains the causes. For this purpose, a retrospective study was conducted for 9587 examinations in canines, of which 4.1% and 10.44% were inconclusive and cancelled, respectively. These results are not in line with ROSE, which foresees a total rate of 5% for these occurrences. The reasons for these high rates in the outpatient clinic were the lack of a Standard Operating Procedure (SOP) and the relative inexperience of the residents in a university hospital. Therefore, with the adoption of the ROSE protocol for veterinary cytopathological examinations, together with adequate training for the outpatient professionals responsible for the examinations, the goals set forth by the ROSE protocol would be met and the rates of inconclusive and cancelled examinations would decrease considerably. Lastly, this study hopes to contribute towards the diagnosis and therapeutic protocols of the main diseases affecting dogs.
文摘Milk production in Ecuador has enormous economic importance and large-, medium- and small-scale producers all participate in the market. There are multiple climatic regions, and dairy production is present in every one of them. High ambient temperatures in the Ecuadorian tropics represent a key challenge to the conservation of milk in the custody of smallholders. The objective of the present study was to evaluate the efficiency of the application of a chemical activator of the Lactoperoxidase System (LP-s) in the conservation of raw milk, at room temperature, in the Ecuadorian tropics. In the present study, sodium thiocyanate—0.36 g·L-1 of milk—and sodium percarbonate—1.36 g·L-1 of milk—as an activator of LP-s were used and the pH and microbiological characteristics (total coliforms, Staphylococcus aureus, total aerobes, molds and yeasts) of the milk at different storage times (0, 4 and 8 hours). The results obtained in the present study showed a significant difference between the two groups under study at 8 hours of storage at room temperature in all parameters (except yeasts where there was no growth in the two treatments), being relevant the significant decrease of the bacterial content. Thus the present study shows that the use of sodium thiocyanate and sodium percarbonate in the above described concentrations could be modulating the activation of LP-s that provides an efficient alternative for the conservation of the raw milk without refrigeration, improving the income for losses of the product and obtaining a raw material of good quality for sale or for further processing, mainly for small producers who do not have the economic resources to have refrigeration means for their product and who must transport their milk for considerably longer distances until they arrive at the collection centers or the processing plants for sale, thus showing that the method used in the present study is not only effective but also has a relatively low cost and easy application.
文摘Milk is one of the products that can be adulterated in many ways affecting the quality of this and its derivatives. Glucomacropeptide (GMP) is a protein that is found only in the whey from the production of fresh cheese, enzymatically obtained from the coagulation of casein and which is commonly used to adulterate fresh or powdered milk. The aim of this study was to determine the adulteration of milk with cheese whey thought a molecular approach, where the glucomacropeptide was collected by sequential precipitation with trichloroacetic acid (ATC) and detected by polyacrylamidododecylsulfate gel electrophoresis (PAGE-SDS), using samples of fresh milk, intentionally adulterated with serum in the proportion of 0%, 1%, 5%, 10% and 15%. The results obtained showed that the detection of glucomacropeptide by electrophoresis was positive in all samples of adulterated milk, evidencing a band of 20.9 kDa in the reading, corresponding to the molecular weight of the GMP, showing that the technique used determines the adulteration in the milk, in a specific and sensitive way, also shows that in the evaluation of physical-chemical and microbiological parameters of milk, there are no significant differences between treatments, except for the pH that tends to decrease as the percentage of serum in the milk increases.
文摘The sarcoma is the generic nomenclature for neoplasm of mesodermal cells, which express in man and animals. Silent growth requires early diagnosis technique for identifying their proteins. The experimental model in vivo murine sarcoma 180-TG (TG-180), is widely used in research to provide the stimuli of infectious and neoplastic antigens. In this case, the technique of immunohisto-chemistry helps identify the expressions of tumor cell variants. The objective of the research was to characterize immunoexpression murine sarcoma TG 180, by immunohistochemistry, antibodies AE1/AE3, vimentin, CD3, CD 45, CD79α and S100A4. For this, murine sarcoma TG-180, was implanted subcutaneously in 20 mice “Swiss”, male, 30 days old, 28 g for 10 days. Samples were taken and subjected to immunohistochemistry, via use of HistoMouse-MA™? kit. There was specifically labeled S100A4 and vimentin antibodies, indicative of poorly differentiated neoplasms fibroblasts. In fact, the model is established by identifying the origin of the cell, once identified, chemotherapeutic tests can also be performed. Neoplasia like these, when installed in man and animals, depending on the degree of aggressiveness requires treatment protocol varied between surgery and chemotherapy or combination of treatments.
基金the Sao Paulo Research Foundation(FAPESP),Brazil,which provided financial support for Francisco Pedraza-Ordonez through the Post-doctoral Fellowship2011/23737-3.
文摘Background: Several animal models, including dogs, have been useful to compare the pathogenesis of mammary neoplasm in humans, showing biological parallelism in the growth and development of breast cancer. The causes of cancer could be attributed to change in several tumor suppressor genes. The relationship between molecule associated to senescence and clinical prognosis of patients affected by mammary cancer is little known. Beyond a collection of data, the major interest of the present study was to carry out a clinical follow-up of patients affected by these tumors, through association with new molecular markers by immunohistochemical technic. Upon completing the study, 15 patients survived, while 45 died. In the case of malignant neoplasms, 40 patients died because of the illness. The type of surgery most used by veterinarian surgeons was the simple lumpectomy, followed by the regional mastectomy. Sentinel node was removed by surgery only when clearly affected. Result: Markings against steroid hormones were positive. Regarding the markings against HER2 and Ki-67, they were negative in all cases. The markings against P53 and CD31 were all positives. Markings against molecules associated with cellular senescence were all positives. No statistical differences were found in immunomarcation for the different antigens used as clinical prognosis factors in mammary neoplasms. Conclusions: According to the study conditions, the survival of patients affected by breast tumors is directly related to diagnosis and malignancy histological grade, but not to animal breed, number of affected glands or patient reproductive status. On the other hand, immunohistochemical markings were not related to the patient prognosis. For this reason, it is important to highlight the persistance of a high percentage of mammary neoplasm cases clinically diagnosed with poor results on patient survival. Thus, educating owners and veterinarians for using diagnostic available tools to improve the prognosis after surgical animals affected by breast cancer is quite necessary.
基金FAPESP(2009/53493-9)and(2009/53777-7)for the financial support
文摘Background: Experimental studies have shown that cyclo-oxygenase-2 (Cox2) is related to the development and progression of tumors, since this enzyme is induced and expressed by cells such as macrophages, osteoblasts, “activated” endothelial cells, and tumor cells. The activity in tumors includes proliferation, cell transformation, tumor growth, invasion and metastasis and may play an important role in carcinogenesis of the canine osteosarcoma, since it has high expression in tissue fragments. The combination of selective Cox2 inhibitors and other treatment modalities is the basis for a new anti-cancer therapy strategy. This in vitro study exposed primary cells of five different canine osteosarcoma cultures to selective Cox2 inhibitor at increasing concentrations and times. Results: For Cox2 negative cultures, despite the absence of differences, greater sensitivity of cells to treatment was observed. For Cox2 positive cultures, a higher number of necrotic cells were observed (P ≤ 0.05), when compared with negative cultures. For exposure times with Celecoxib doses, no difference (P > 0.05) was found between the three times analyzed for living, apoptotic and apoptotic/necrotic cells. There are similarities in the values of 24 h and 48 h, with slight reduction of living cells, increasing those undergoing apoptosis and apoptosis/necrosis. There was significance for necrosis (P ≤ 0.05). In 72 hours, a significant difference was observed between the other two previous values (P ≤ 0.05). It was found for the group of 100 μML-1, that there was a numerically greater signaling for apoptosis and lower (P = 0.08) for necrosis, and this point was the onset of the pharmacodynamic phenomenon, with drop in the values for living cells and increased number of necrotic cells, with a tendency (P = 0.08) for reducing the percentage of necrotic cells for the group of 100 μML-1 when compared to that of 10 μML-1. Conclusions: For Cox2 positive and negative cultures, there was difference for necrotic cells and there was no difference between Cox2 positive and Cox2 negative groups in relation to the percentage of living cells and apoptotic and apoptotic/necrotic cells. At time of 72 hours, higher percentage of living cells, lower percentage of apoptotic cells and increased percentage of necrotic cells in relation to groups of 24 and 48 hours was observed. A tendency for reducing the percentage of necrotic cells for the group of 100 μML-1 when compared to that of the group of 10 μML-1 was observed.
文摘Diagnosis and biological behavior of breast cancer of female dog represent one of the biggest challenges facing the Veterinarian in recent years. Due to its exponential growth and the degree of aggressiveness, the exact cause of this tumor is probably multifactorial and it is believed that may suffer influence from environmental factors. Among the suspected environmental contaminants are the pyrethroids. Aiming to investigate the participation of pyrethroids in tumorigenesis in female dogs, a study was conducted using 50 female dogs, 22 were positive for simple breast carcinoma (Group I), 18 with a diagnosis of complex breast carcinoma (Group II) and 10 negative (Group III) for breast cancer. In order to detect DNA damage, the Comet assay was performed on mammary samples of these animals, which also had samples submitted to the technique of High Performance Liquid Chromatography (HPLC), which aimed to quantify the concentration of pyrethroids. The results of HPLC of each animal were compared with those obtained by the Comet assay analysis of variance and the means were compared by the test groups “Student T” at the significance level of p ￡ 0.05. Despite presenting correlation between the amount of DNA damage and tumor aggressiveness, no statistical differences were found in the DNA damage of different histologic types of breast carcinoma. As for pyrethroids, even these were detected in 22% of tumor tissues and peritumoral fat, there was no difference in DNA damage between cells exposed and not exposed to environmental contaminant.
文摘Objective: To investigate the potential of superparamagnetic iron oxide particles (SPIO) in MR imaging for the differentiation between hyperplastic and metastatic lymph node. Methods: Animal models of malignant lymph node metastasis were established in 6 New-Zealand rabbits by a unilateral intra-muscular injection of VX2 carcinoma cells, and models of hyperplastic lymph nodes were induced in another 6 rabbits by a unilateral intra-muscular injection of egg yolk emulsion. MR images of the lymph nodes were obtained before and 12 h after interstitial injection of SPIO. Image results were analyzed and compared with pathological findings. Results: On unenhanced images, the signal intensity of hyperplastic and metastatic lymph nodes did not differ significantly. After administration of SPIO, the signal intensity of both hyperplastic and metastatic lymph nodes remained unchanged on T1-weighted SE images. On T2-weighted SE images, the signal intensity of hyperplastic lymph nodes decreased heterogeneously, while that of all metastatic ones remained unchanged. On T2-weighted GRE images, the signal intensity of hyperplastic lymph nodes decreased significantly and homogeneously, while that of 4 metastatic ones remained unchanged and that of the rest 2 decreased heterogeneously. Conclusion: SPIO-enhanced MR imaging may enable the differentiation between the hyperplastic and metastatic lymph nodes.
基金funded by the Else Kröner Fresenius-Stiftung(2019_A149 and 2022_EKMS.26 to J.C.F.)the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)-Projektnummer 360372040-SFB 1335(to F.B.,H.P.,S.H.,and K.S.)+12 种基金Projektnummer 395357507-SFB 1371(P04 to D.H.B.and E.H.,P05 to H.P.,P11 to M.T.and J.C.F.,Z02 to K.S.,seed funding to E.T.O.)Projektnummer 324392634-TRR 221(to H.P.,E.H.,M.B-H.,M.R.,M.F.,P.H.,M.E.,D.W.,and W.H.)Projektnummer BA 2851/6-1(to F.B.)Projektnummer PO 1575/5-1(to H.P.)Projektnummer 509149993-TRR 374(to E.V.,M.B-H.)Bavarian Cancer Research Centre(BZFK,to H.P.,W.H.,and F.B.)the German Cancer Aid(70114547 to H.P.,70113964 to J.C.F.)the Wilhelm Sander Foundation(2023.072.1 to J.C.F.,2021.041.1 to S.H.,2021.040.1 to H.P.)the European Hematology Association(to H.P.)a Mechtild Harf Research Grant from the DKMS Foundation for Giving Life(to H.P.)a Young Investigator Award by the Melanoma Research Alliance(to S.H.)the German JoséCarreras leukemia foundation(DJCLS 07 R/2020 to S.H.)funded/co-funded by the European Union(project MICROBOTS,Grant No.101124680 to H.P.).
文摘Intestinal stem cells(ISCs)promote tissue repair after genotoxic or immune-mediated injury.However,ISCs are particularly sensitive to various stressors and primary targets of overwhelming immune responses,such as interferonγ(IFNγ)-mediated killing.In mouse models of radiation therapy-induced gut damage and in biopsies from patients who underwent allogeneic hematopoietic stem cell transplantation,we observed IFNγexpression by intestinal Treg cells.Treg cells leverage combined IFNγand interleukin 10(IL-10)stimulation of ISCs to nurture the growth of intestinal organoids through the activation of the mTORC1 and Myc pathways.Similarly,Treg cells or the combined addition of recombinant IFNγand IL-10 promoted the regeneration of organoids after irradiation,and both cytokines were essential for ensuring epithelial regeneration following acute intestinal tissue injury in vivo.The exposure of organoids to growth factor-free culture conditions revealed distinct EGF-like properties of IFNγand Wnt-like properties of IL-10.While IFNγrapidly induced epithelial proliferation,it depleted the pool of ISCs in vitro.Only the combination of IFNγand IL10 led to epithelial proliferation and organoid growth while simultaneously ensuring ISC maintenance over time.Our results reveal a context-dependent role of inflammatory signaling in ISCs,through which Treg cells promote epithelial repair following therapyinduced injury.
文摘Halogenated natural products(HNPs)are considered to be emerging contaminants whose environmental distribution and fate are only incompletely known.Therefore,several persistent and bioaccumulative HNP groups,together with manmade polychlorinated biphenyls(PCBs)and polybrominated diphenyl ethers(PBDEs),were quantified in the blubber of nine sperm whales(Physeter macrocephalus)stranded on the coast of the Mediterranean Sea in Italy.The naturally occurring polybrominated hexahydroxanthene derivatives(PBHDs;sum of TetraBHD and TriBHD)were the most prominent substance class with up to 77,000 ng/g blubber.The mean PBHD content(35,800 ng/g blubber)even exceeded the one of PCBs(28,400 ng/g blubber),although the region is known to be highly contaminated with manmade contaminants.Based on mean values,Q1∼PBDEs>MeO-BDEs∼2,2′-diMeO-BB 80 and several other HNPs followed with decreasing amounts.All blubber samples contained an abundant compound whose molecular formula(C_(16)H_(19)Br_(3)O_(2))was verified using high-resolution mass spectrometry.The only plausible matching isomer was(2S,4′S,9R,9′S)-2,7-dibromo-4′-bromomethyl-1,1-dimethyl-2,3,4,4′,9,9′-9,9′-hexahydro-1H-xanthen-9-ol(OH-TriBHD),a hydroxylated secondary metabolite previously detected together with TriBHD and TetraBHD in a sponge known to be a natural producer of PBHDs.The estimated mean amount of the presumed OH-TriBHD was 3000 ng/g blubber,which is unexpectedly high for hydroxylated compounds in the lipids of marine mammals.
