Introduction The current 30% of canine perinatal mortality (Tonnessen et al., 2012) claims the need to deepen the knowledge about this phase, characterized by long-term hormonal and metabolic changes, that could benefit of long time-frame methods of investigation, reducing the quantity of samplings and respecting animal welfare (Veronesi et al., 2015). Therefore, blood, saliva, urine and feces, being invasive and providing punctual information, are not appropriate matrices. Given that sexual steroid hormones were reported to have an influence already from the perinatal period (Frey et al., 2017), in this work 17-β-Estradiol (E2) and Testosterone (T) concentrations were assessed from the claws of dogs up to 60 days of age. Material and Methods Ten male and 10 female puppies, viable and healthy, born by elective cesarean section, were enrolled. Samplings were performed by trimming claws at birth, and the regrowth at 30 and 60 days of age. Then, E2 and T concentrations were analyzed by RIA (Veronesi et al., 2015) and a possible effect of gender evaluated. Results All the hormonal claws concentrations showed a significant (p<0.001) drop from birth to 30 and 60 days of age, while no significant changes were observed between 30 and at 60 days of age (Table 1). No influence of the newborns’ gender was found. Discussion and Conclusions Because of the high levels of E2 and T observed at birth and at 30 days of age, it could be supposed that a source of production could be the placental and maternal compartments. However, the direct involvement of the fetus itself could not be excluded, given the reported accumulation from the nail capillary bed of those hormones (de Berker et al., 2007) and the production of sexual steroid hormones by feline fetal gonads (Braun and Jewgenow, 2017).
目前30%的犬类围产期死亡率(Tonnessen et al.， 2012)声称需要加深对这一阶段的了解，这一阶段的特征是长期的激素和代谢变化，这可能有利于长期的调查方法，减少采样数量并尊重动物福利(Veronesi et al.， 2015)。因此，血液、唾液、尿液和粪便具有侵入性并能及时提供信息，不适合作为基质。鉴于据报道，性类固醇激素从围产期就已经有影响(Frey et al.， 2017)，在本工作中，从60日龄的狗爪开始评估17-β-雌二醇(E2)和睾酮(T)浓度。材料与方法选择择期剖宫产出生的健康存活幼犬公、母各10只。出生时剪爪，30日龄和60日龄再生。然后，通过RIA分析E2和T浓度(Veronesi et al.， 2015)，并评估性别可能的影响。结果从出生到30和60日龄，所有激素爪浓度均显著(p<0.001)下降，而30和60日龄之间无显著变化(表1)。未发现新生儿性别的影响。讨论和结论由于在出生和30日龄时观察到E2和T的高水平，可以推测产生E2和T的来源可能是胎盘和母体间室。然而，不能排除胎儿本身的直接参与，因为有报道称这些激素在指甲毛细血管床上积累(de Berker等人，2007)，而且猫胎儿性腺会产生性类固醇激素(Braun和Jewgenow, 2017)。
{"title":"17-ß-Estradiol and Testosterone concentrations in claws of puppies up to 60 days of age","authors":"J. Fusi, T. Peric, Cristina Bergamin, M. Veronesi","doi":"10.13130/2283-3927/9992","DOIUrl":"https://doi.org/10.13130/2283-3927/9992","url":null,"abstract":"Introduction The current 30% of canine perinatal mortality (Tonnessen et al., 2012) claims the need to deepen the knowledge about this phase, characterized by long-term hormonal and metabolic changes, that could benefit of long time-frame methods of investigation, reducing the quantity of samplings and respecting animal welfare (Veronesi et al., 2015). Therefore, blood, saliva, urine and feces, being invasive and providing punctual information, are not appropriate matrices. Given that sexual steroid hormones were reported to have an influence already from the perinatal period (Frey et al., 2017), in this work 17-β-Estradiol (E2) and Testosterone (T) concentrations were assessed from the claws of dogs up to 60 days of age. Material and Methods Ten male and 10 female puppies, viable and healthy, born by elective cesarean section, were enrolled. Samplings were performed by trimming claws at birth, and the regrowth at 30 and 60 days of age. Then, E2 and T concentrations were analyzed by RIA (Veronesi et al., 2015) and a possible effect of gender evaluated. Results All the hormonal claws concentrations showed a significant (p<0.001) drop from birth to 30 and 60 days of age, while no significant changes were observed between 30 and at 60 days of age (Table 1). No influence of the newborns’ gender was found. Discussion and Conclusions Because of the high levels of E2 and T observed at birth and at 30 days of age, it could be supposed that a source of production could be the placental and maternal compartments. However, the direct involvement of the fetus itself could not be excluded, given the reported accumulation from the nail capillary bed of those hormones (de Berker et al., 2007) and the production of sexual steroid hormones by feline fetal gonads (Braun and Jewgenow, 2017).","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84898945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10013
F. Genova, M. Longeri, Cristina M Cozzi, A. Bagnato, M. Strillacci
Copy Number Variations (CNVs) have become promising markers, representing a major source of genomic variation. CNV involvement in phenotypic expression and in different diseases onset have been widely demonstrated in humans as well as in many domestic animals. However, this genomic investigation is still missing in Felis catus. This work is the first CNV mapping from a large data set of Whole Genome Sequencing (WGS) data in the domestic cat. A total of 42 cats of 14 different breeds were sequenced on the Illumina XTen (Washington University-St. Louis) which generated approximately 30-fold genome coverage from 150 paired-end reads (99 Lives Initiative). Maverix Biomics mapped the reads on the v6.2 reference assembly. CNV detection was performed using cn.mops and CNVnator, two Read Depth method software. One cat was excluded as outlier while, on the 41 remaining individuals, 1640 CNVs were detected by both the software and used to obtain 2891 CNVRs with BedTools. CNVRs covered the 0.4% of the total cat genome, with 136 loss, 127 gain and 26 complex detected (Fig. 1). A total of 164 singletons were identified and 9 CNVRs mapped in at least the 50% of the individuals. The number of CNVs in each cat ranged from 12 to 83. The clustering analysis of the detected CNVs was performed with R package “pvclust” and shows that same breed individuals cluster together. This study has led to the genetic characterization of 14 main cat breeds. Further analyses including other breeds and considering the genes located within these regions, could led to better evaluate the relationship between the presence of a specific CNV and a specific breed trait. This study can be considered a starting point for genomic CNV identification in the domestic cat, which could be further developed using the new released Felis catus vs9.0 reference aassembly.
拷贝数变异(拷贝数变异)作为基因组变异的主要来源，已成为一种很有前景的标记。CNV参与表型表达和不同疾病的发病已在人类和许多家畜中得到广泛证实。然而，这种基因组调查在猫科动物中仍然缺失。这项工作是第一个来自家猫全基因组测序(WGS)数据的大型数据集的CNV图谱。在Illumina XTen(华盛顿大学- st .)上对14个不同品种的42只猫进行了测序。Louis)从150对末端读取产生了大约30倍的基因组覆盖率(99 Lives Initiative)。Maverix Biomics将读取映射到v6.2参考程序集上。CNV检测使用cn。拖把和CNVnator，两个读深法软件。一只猫被排除为异常值，而在剩余的41只猫中，两种软件检测到1640个cnvr，并使用BedTools获得2891个cnvr。cnvr覆盖了猫总基因组的0.4%，其中检测到136个缺失，127个获得和26个复合体(图1)。总共鉴定出164个单子，并在至少50%的个体中绘制了9个cnvr。每只猫的CNVs数量从12到83不等。利用R软件包“pvclust”对检测到的CNVs进行聚类分析，发现同一品种的个体聚类在一起。这项研究得出了14个主要猫品种的遗传特征。进一步的分析包括其他品种并考虑这些区域内的基因，可以更好地评估特定CNV的存在与特定品种性状之间的关系。本研究可视为家猫基因组CNV鉴定的一个起点，可使用新发布的Felis catus vs9.0参考汇编进一步发展。
{"title":"Genome-wide CNV Mapping in Felis catus Using NGS Data","authors":"F. Genova, M. Longeri, Cristina M Cozzi, A. Bagnato, M. Strillacci","doi":"10.13130/2283-3927/10013","DOIUrl":"https://doi.org/10.13130/2283-3927/10013","url":null,"abstract":"Copy Number Variations (CNVs) have become promising markers, representing a major source of genomic variation. CNV involvement in phenotypic expression and in different diseases onset have been widely demonstrated in humans as well as in many domestic animals. However, this genomic investigation is still missing in Felis catus. This work is the first CNV mapping from a large data set of Whole Genome Sequencing (WGS) data in the domestic cat. A total of 42 cats of 14 different breeds were sequenced on the Illumina XTen (Washington University-St. Louis) which generated approximately 30-fold genome coverage from 150 paired-end reads (99 Lives Initiative). Maverix Biomics mapped the reads on the v6.2 reference assembly. CNV detection was performed using cn.mops and CNVnator, two Read Depth method software. One cat was excluded as outlier while, on the 41 remaining individuals, 1640 CNVs were detected by both the software and used to obtain 2891 CNVRs with BedTools. CNVRs covered the 0.4% of the total cat genome, with 136 loss, 127 gain and 26 complex detected (Fig. 1). A total of 164 singletons were identified and 9 CNVRs mapped in at least the 50% of the individuals. The number of CNVs in each cat ranged from 12 to 83. The clustering analysis of the detected CNVs was performed with R package “pvclust” and shows that same breed individuals cluster together. This study has led to the genetic characterization of 14 main cat breeds. Further analyses including other breeds and considering the genes located within these regions, could led to better evaluate the relationship between the presence of a specific CNV and a specific breed trait. This study can be considered a starting point for genomic CNV identification in the domestic cat, which could be further developed using the new released Felis catus vs9.0 reference aassembly.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88627149","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10000
L. Nordio, S. Valentina, C. Giudice
Feline diffuse iris melanoma (FDIM) is the most common primary intraocular neoplasm in cats. It is usually a malignant tumor, even if slowly progressive, thus representing an unique spontaneous model of the aggressive, although rare, human iris melanoma. In cats, the extent of the tumor within the eye, expressed as histological grade, is considered a good predictor of survival. In the context of the neoplastic cells-tumor microenvironment interaction, Matrix Metalloproteinase-9 (MMP-9) is an endopeptidase able to digest the extracellular matrix with involvement in tumor invasion . MMP-9 expression has been positively correlated with metastasizing behavior in human posterior uveal melanoma. The present study investigates the expression of MMP-9 in a caseload of formalin-fixed paraffin-embedded FDIMs in relation to the histological grade and mitotic index (MI) (threshold=7/10 hpf). Sixty-one samples of FDIM evaluated on light microscopy (Fig. 1) were selected (grade I n=22, grade II n=20, grade III n=19). Immunohistochemical staining with standard ABC method was performed using a mouse anti-MMP-9 antibody. Results were semi-quantitatively scored and compared by Mann-Whitney U test. MMP-9 was expressed in 59,1% grade I FDIM, 90,0% grade II and 80,0% grade III. Tumors with MMP-9 expression in more than 50% of neoplastic cells were 13,6% in grade I cases, 40,0% in grade II and 36,8% in grade III. MMP-9 was expressed in 71,4% of FDIM with MI≤7 and 92,3% of FDIM with MI>7. MMP-9 expression differed significantly between grade I and the other two grades, and between groups with low and high MI. In conclusion, intense expression of MMP-9 seems to correlate with the histological aggressiveness of FDIM.
{"title":"MMP-9 immunohistochemical expression is correlated with histologic grade in feline diffuse iris melanoma","authors":"L. Nordio, S. Valentina, C. Giudice","doi":"10.13130/2283-3927/10000","DOIUrl":"https://doi.org/10.13130/2283-3927/10000","url":null,"abstract":"Feline diffuse iris melanoma (FDIM) is the most common primary intraocular neoplasm in cats. It is usually a malignant tumor, even if slowly progressive, thus representing an unique spontaneous model of the aggressive, although rare, human iris melanoma. In cats, the extent of the tumor within the eye, expressed as histological grade, is considered a good predictor of survival. In the context of the neoplastic cells-tumor microenvironment interaction, Matrix Metalloproteinase-9 (MMP-9) is an endopeptidase able to digest the extracellular matrix with involvement in tumor invasion . MMP-9 expression has been positively correlated with metastasizing behavior in human posterior uveal melanoma. The present study investigates the expression of MMP-9 in a caseload of formalin-fixed paraffin-embedded FDIMs in relation to the histological grade and mitotic index (MI) (threshold=7/10 hpf). Sixty-one samples of FDIM evaluated on light microscopy (Fig. 1) were selected (grade I n=22, grade II n=20, grade III n=19). Immunohistochemical staining with standard ABC method was performed using a mouse anti-MMP-9 antibody. Results were semi-quantitatively scored and compared by Mann-Whitney U test. MMP-9 was expressed in 59,1% grade I FDIM, 90,0% grade II and 80,0% grade III. Tumors with MMP-9 expression in more than 50% of neoplastic cells were 13,6% in grade I cases, 40,0% in grade II and 36,8% in grade III. MMP-9 was expressed in 71,4% of FDIM with MI≤7 and 92,3% of FDIM with MI>7. MMP-9 expression differed significantly between grade I and the other two grades, and between groups with low and high MI. In conclusion, intense expression of MMP-9 seems to correlate with the histological aggressiveness of FDIM.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73108307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10033
E. Mariani, G. Savoini, T. Niewold
Feed additives are commonly used to improve pig performance and health, but they need to be tested so new biomarkers for intestinal health, non- or minimally invasive, are under investigations.The quantification of Myeloperoxidase (MPO) and Pancreatitis Associated Protein (PAP) in feces could prove useful to non-invasively monitor intestinal health (Niewold, 2015). MPO is an enzyme that permits to quantify the number of inflammatory cells present in tissues and feces (Prokopowicz et al., 2012) , while PAP is a protein mainly produced in the small intestine with anti-inflammatory and bactericidal activity (Cash et al., 2006; Mukherjee et al., 2014). Because of the lack of a commercial ELISA kit for porcine PAP detection, the main aim of this study was to develop and validate a new sandwich ELISA test for the quantification of PAP in pig fecal samples. Our study consisted of two phases: test development and test validation. During the development phase we used polyclonal antibodies previously immunized from rabbit serum with a pure peptide containing the N-terminus of pig PAP (Soler et al., 2015). The validation of the test was then performed with fecal extraction samples derived from animals with known high or low growth performance.Moreover, the temperature stability of PAP in feces and the optimal extraction method was tested. Even if only preliminary, our results seem to show a fair relationship between fecal consistency, used as health indicator, and PAP fecal concentrations. Furthermore, no relevant differences in PAP concentration after 24h of incubation at 37 °C, 4°C or room temperature were detected.To date, the present results suggest that PAP seems to be exceptionally stable in feces and is a very promising candidate as a non-invasive (fecal) biomarker for intestinal health and growth.
饲料添加剂通常用于改善猪的生产性能和健康，但它们需要进行测试，因此正在研究新的肠道健康生物标志物，非侵入性或微创性。粪便中髓过氧化物酶(MPO)和胰腺炎相关蛋白(PAP)的定量可能有助于无创监测肠道健康(Niewold, 2015)。MPO是一种可以量化组织和粪便中炎症细胞数量的酶(Prokopowicz et al.， 2012)，而PAP是一种主要产生于小肠的蛋白质，具有抗炎和杀菌活性(Cash et al.， 2006;Mukherjee et al.， 2014)。由于缺乏用于猪PAP检测的商用ELISA试剂盒，本研究的主要目的是开发和验证一种新的夹心ELISA检测方法，用于猪粪便样品中PAP的定量。我们的研究包括两个阶段:测试开发和测试验证。在开发阶段，我们使用了先前从兔血清中免疫的多克隆抗体，其中含有猪PAP的n端纯肽(Soler et al.， 2015)。然后用从已知生长性能高或低的动物身上提取的粪便样本对试验进行验证。并对粪便中PAP的温度稳定性和最佳提取方法进行了试验。即使只是初步的，我们的结果似乎显示了作为健康指标的粪便稠度与PAP粪便浓度之间的公平关系。此外，在37°C、4°C和室温条件下孵育24h后，PAP浓度无相关差异。到目前为止，目前的结果表明PAP在粪便中似乎非常稳定，是一个非常有前途的肠道健康和生长的非侵入性(粪便)生物标志物。
{"title":"Non-invasive intestinal biomarkers: a new ELISA test for Pancreatitis Associated Protein detection in pig","authors":"E. Mariani, G. Savoini, T. Niewold","doi":"10.13130/2283-3927/10033","DOIUrl":"https://doi.org/10.13130/2283-3927/10033","url":null,"abstract":"Feed additives are commonly used to improve pig performance and health, but they need to be tested so new biomarkers for intestinal health, non- or minimally invasive, are under investigations.The quantification of Myeloperoxidase (MPO) and Pancreatitis Associated Protein (PAP) in feces could prove useful to non-invasively monitor intestinal health (Niewold, 2015). MPO is an enzyme that permits to quantify the number of inflammatory cells present in tissues and feces (Prokopowicz et al., 2012) , while PAP is a protein mainly produced in the small intestine with anti-inflammatory and bactericidal activity (Cash et al., 2006; Mukherjee et al., 2014). Because of the lack of a commercial ELISA kit for porcine PAP detection, the main aim of this study was to develop and validate a new sandwich ELISA test for the quantification of PAP in pig fecal samples. Our study consisted of two phases: test development and test validation. During the development phase we used polyclonal antibodies previously immunized from rabbit serum with a pure peptide containing the N-terminus of pig PAP (Soler et al., 2015). The validation of the test was then performed with fecal extraction samples derived from animals with known high or low growth performance.Moreover, the temperature stability of PAP in feces and the optimal extraction method was tested. Even if only preliminary, our results seem to show a fair relationship between fecal consistency, used as health indicator, and PAP fecal concentrations. Furthermore, no relevant differences in PAP concentration after 24h of incubation at 37 °C, 4°C or room temperature were detected.To date, the present results suggest that PAP seems to be exceptionally stable in feces and is a very promising candidate as a non-invasive (fecal) biomarker for intestinal health and growth.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78359701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10017
U. Polito, M. Giancamillo, G. Peretti, M. E. Andreis, S. Modina, F. Boschetti, A. Giancamillo
Menisci are wedge-like structures interposed, in the knee joint, between the femoral and the tibial articular heads (Kohn et al. 1995; Greis et al. 2002). Improving the articular surface, the cellular nutrition and the articular lubrication, they are essential structures for the prevention of gonarthrosis (Proctor et al.1989; Makris et al. 2011). This study is focused upon the relationship between the contact forces at the femoral and tibial surfaces and the corresponding structure of these meniscal surfaces. For this purpose, 20 adult (~9 months old) female pigs (Landrace x Large white, average weight 75–90 kg; n=80 meniscal samples) were obtained from a local slaughterhouse and dissected to isolate the menisci. Swine meniscal samples were evaluated from morphological (Safranin-O, Sirius Red and collagen type I and II) (Di Giancamillo et al. 2014), biochemical (DNA and glycosaminoglycans, or GAGs, contents) and biomechanical (compression and traction tests) points of view at the level of femoral and tibial meniscal surfaces. Results revealed a characterization of the meniscus which is biomechanical-dependent. The femoral surface, morphologically characterized by the interposition of radial and oblique fibers and biomechanically by the femoral condyles compression, sliding and rolling forces, shows a higher compressive modulus (p<0.05) and a greater amount of cells and GAGs deposition (p<0.01 for each analysis). On the other hand, results from traction test revealed a higher tensile modulus (p<0.05) in the tibial surface, characterized by a circumferential arrangement of the fibers and a poorer GAGs deposition and cellular distribution (p<0.01). Results (summarized in the figure 1) from this work suggest that a biphasic “femoral-to-tibial” scaffold that mimic the different behavior and composition of the two meniscal surfaces could be useful in the light of meniscal replacement.
半月板是位于膝关节股骨和胫骨关节头之间的楔形结构(Kohn et al. 1995;Greis et al. 2002)。改善关节表面，细胞营养和关节润滑，它们是预防关节病必不可少的结构(Proctor等，1989;Makris et al. 2011)。本研究的重点是股骨和胫骨表面的接触力与这些半月板表面的相应结构之间的关系。为此，选用20头成年(~9月龄)母猪(长白猪x大白猪，平均体重75-90公斤;N =80半月板样本)从当地屠宰场获得并解剖分离半月板。猪半月板样本在股骨和胫骨半月板表面水平从形态学(红花素- o、Sirius Red和I型和II型胶原蛋白)(Di Giancamillo等人，2014年)、生化(DNA和糖胺聚糖，或GAGs，含量)和生物力学(压缩和牵引试验)角度进行评估。结果显示半月板的特征是生物力学依赖的。股骨表面形态特征为径向和斜向纤维的插入，生物力学特征为股骨髁的压缩、滑动和滚动力，其压缩模量较高(p<0.05)，细胞和GAGs沉积量较高(p<0.01)。另一方面，牵引试验结果显示胫骨表面的拉伸模量较高(p<0.05)，纤维呈周向排列，GAGs沉积和细胞分布较差(p<0.01)。这项工作的结果(总结在图1中)表明，模拟两个半月板表面的不同行为和组成的双相“股胫”支架在半月板置换术中可能是有用的。
{"title":"Meniscal femoral and tibial surfaces characterization in the swine model.","authors":"U. Polito, M. Giancamillo, G. Peretti, M. E. Andreis, S. Modina, F. Boschetti, A. Giancamillo","doi":"10.13130/2283-3927/10017","DOIUrl":"https://doi.org/10.13130/2283-3927/10017","url":null,"abstract":"Menisci are wedge-like structures interposed, in the knee joint, between the femoral and the tibial articular heads (Kohn et al. 1995; Greis et al. 2002). Improving the articular surface, the cellular nutrition and the articular lubrication, they are essential structures for the prevention of gonarthrosis (Proctor et al.1989; Makris et al. 2011). This study is focused upon the relationship between the contact forces at the femoral and tibial surfaces and the corresponding structure of these meniscal surfaces. For this purpose, 20 adult (~9 months old) female pigs (Landrace x Large white, average weight 75–90 kg; n=80 meniscal samples) were obtained from a local slaughterhouse and dissected to isolate the menisci. Swine meniscal samples were evaluated from morphological (Safranin-O, Sirius Red and collagen type I and II) (Di Giancamillo et al. 2014), biochemical (DNA and glycosaminoglycans, or GAGs, contents) and biomechanical (compression and traction tests) points of view at the level of femoral and tibial meniscal surfaces. Results revealed a characterization of the meniscus which is biomechanical-dependent. The femoral surface, morphologically characterized by the interposition of radial and oblique fibers and biomechanically by the femoral condyles compression, sliding and rolling forces, shows a higher compressive modulus (p<0.05) and a greater amount of cells and GAGs deposition (p<0.01 for each analysis). On the other hand, results from traction test revealed a higher tensile modulus (p<0.05) in the tibial surface, characterized by a circumferential arrangement of the fibers and a poorer GAGs deposition and cellular distribution (p<0.01). Results (summarized in the figure 1) from this work suggest that a biphasic “femoral-to-tibial” scaffold that mimic the different behavior and composition of the two meniscal surfaces could be useful in the light of meniscal replacement.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80098593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10011
B. Ruggerone, R. Troìa, E. Murgia, M. Giunti, F. Dondi, S. Paltrinieri
An early diagnosis of sepsis could allow a better prognosis and avoid the abuse of antibiotic administration; unfortunately, in veterinary medicine, specific and sensitive markers of sepsis are not available.Because Protein Carbonyls (PCOs), that result from protein oxidation, are widely used in human medicine as sepsis markers , the aim of our study was to validate an ELISA kit (Enzolifesciences, 3V Chimica, Roma) on canine serum and to measure PCOs, after a preliminary validation study, in three groups (homogeneous for age and size): healthy dogs without clinical or laboratory abnormalities (A, n=14), dogs with septic (B, n=14) and non-septic inflammation (C, n=12) at the first presentation and without previous treatments. Moreover, Paraoxonase-1, a negative acute phase protein with anti-oxidant properties (PON-1) was measured in each group with a method already validated in dogs.A Kruskal-Wallis test followed by a Wilcoxon signed rank test was used to evaluate differences between groups.The ELISA method for measuring PCOs showed a very good precision (coefficient of variation <12%) and a good accuracy in spiking-recovery tests.Compared with controls, the concentration of PCOs was significantly higher either in dogs with sepsis (P<0.001) or in dogs with non-septic inflammation (P=0.005) but no significant differences were found between the two groups of sick dogs. Conversely, PON-1 was significantly lower in sick dogs compared with controls (P<0.001 for both groups) and in septic dogs compared with dogs with non-septic inflammation (P=0.001).A negative correlation between the two markers was found (P<0.001, r=-0.594) Receiver operating characteristic (ROC) curves demonstrated that both markers may discriminate dogs with sepsis with the other groups. However, PCO was less sensitive than PON-1 in diagnosing sepsis.Future studies should be focused on the association of PCOs with other inflammatory markers, as well as the possible prognostic role of PCOs based on the outcome of the enrolled patients.
{"title":"Diagnosis of sepsis in dogs by measuring carbonylated proteins (PCOs) and paraoxonase (PON-1)","authors":"B. Ruggerone, R. Troìa, E. Murgia, M. Giunti, F. Dondi, S. Paltrinieri","doi":"10.13130/2283-3927/10011","DOIUrl":"https://doi.org/10.13130/2283-3927/10011","url":null,"abstract":"An early diagnosis of sepsis could allow a better prognosis and avoid the abuse of antibiotic administration; unfortunately, in veterinary medicine, specific and sensitive markers of sepsis are not available.Because Protein Carbonyls (PCOs), that result from protein oxidation, are widely used in human medicine as sepsis markers , the aim of our study was to validate an ELISA kit (Enzolifesciences, 3V Chimica, Roma) on canine serum and to measure PCOs, after a preliminary validation study, in three groups (homogeneous for age and size): healthy dogs without clinical or laboratory abnormalities (A, n=14), dogs with septic (B, n=14) and non-septic inflammation (C, n=12) at the first presentation and without previous treatments. Moreover, Paraoxonase-1, a negative acute phase protein with anti-oxidant properties (PON-1) was measured in each group with a method already validated in dogs.A Kruskal-Wallis test followed by a Wilcoxon signed rank test was used to evaluate differences between groups.The ELISA method for measuring PCOs showed a very good precision (coefficient of variation <12%) and a good accuracy in spiking-recovery tests.Compared with controls, the concentration of PCOs was significantly higher either in dogs with sepsis (P<0.001) or in dogs with non-septic inflammation (P=0.005) but no significant differences were found between the two groups of sick dogs. Conversely, PON-1 was significantly lower in sick dogs compared with controls (P<0.001 for both groups) and in septic dogs compared with dogs with non-septic inflammation (P=0.001).A negative correlation between the two markers was found (P<0.001, r=-0.594) Receiver operating characteristic (ROC) curves demonstrated that both markers may discriminate dogs with sepsis with the other groups. However, PCO was less sensitive than PON-1 in diagnosing sepsis.Future studies should be focused on the association of PCOs with other inflammatory markers, as well as the possible prognostic role of PCOs based on the outcome of the enrolled patients.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88175759","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10265
M. Bhide
Knowledge of host-pathogen interactions at molecular level is crucial for understanding of pathogenesis, disease prevention and cure. Here Borrelia is presented as a model neuroinvasive pathogen which employs vast immune evasion mechanisms like - drastic change in antigenic proteins, complement regulatory protein binding, antigenic variation etc. It is proposed that Borrelia may employ multiple strategies to evade host's complement system by binding complement regulatory proteins like factor H, vitronectin, C4BP and CD59. Amplitude of these mechanisms, their pathogen species dependent disparity, and characterization of interacting proteins from both sides (host and pathogen) will be presented during VAS presentations. Borrelia is also capable of invading central nervous system (CNS) in hide in the immune privileged site – the CNS. To do so it must cross the blood brain barrier (BBB). The mechanisms of the BBB crossing of this organism, like many of the other CNS invading pathogens, are still subject of on-going research. Our decade of research indicate the paracellular mechanism of the BBB penetration of Borrelia . Paracellular penetration of pathogen needs multiple protein:protein interactions between pathogen surface proteins and endothelial cells. The second part of my presentation in VAS will be dedicated to mechanisms employed by Borrelia to modulate cell signaling events in brain microvascular endothelial cells to cross the BBB. This presentation will also give sneak peak of several state-of-the-art technologies in genomics and proteomic used to understand the mechanisms of neuroinvasive and complement evasion by Borrelia . I hope that doctoral fellows will take advantage of the experimental pipeline that will be presented in VAS and apply successfully in their research
{"title":"Borrelia and host interaction: a tangled tale","authors":"M. Bhide","doi":"10.13130/2283-3927/10265","DOIUrl":"https://doi.org/10.13130/2283-3927/10265","url":null,"abstract":"Knowledge of host-pathogen interactions at molecular level is crucial for understanding of pathogenesis, disease prevention and cure. Here Borrelia is presented as a model neuroinvasive pathogen which employs vast immune evasion mechanisms like - drastic change in antigenic proteins, complement regulatory protein binding, antigenic variation etc. It is proposed that Borrelia may employ multiple strategies to evade host's complement system by binding complement regulatory proteins like factor H, vitronectin, C4BP and CD59. Amplitude of these mechanisms, their pathogen species dependent disparity, and characterization of interacting proteins from both sides (host and pathogen) will be presented during VAS presentations. Borrelia is also capable of invading central nervous system (CNS) in hide in the immune privileged site – the CNS. To do so it must cross the blood brain barrier (BBB). The mechanisms of the BBB crossing of this organism, like many of the other CNS invading pathogens, are still subject of on-going research. Our decade of research indicate the paracellular mechanism of the BBB penetration of Borrelia . Paracellular penetration of pathogen needs multiple protein:protein interactions between pathogen surface proteins and endothelial cells. The second part of my presentation in VAS will be dedicated to mechanisms employed by Borrelia to modulate cell signaling events in brain microvascular endothelial cells to cross the BBB. This presentation will also give sneak peak of several state-of-the-art technologies in genomics and proteomic used to understand the mechanisms of neuroinvasive and complement evasion by Borrelia . I hope that doctoral fellows will take advantage of the experimental pipeline that will be presented in VAS and apply successfully in their research","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87574270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10268
A. Muñoza
Recently, a scientific interest in exercise in water in equine athletes, both from training and rehabilitation purposes has been developed. Before an extended use of this practice in equine exercise physiology and medicine, the physiological adaptations to this type of exercise need to be elucidated. Six horses were subjected to sessions of 40 minutes in water treadmill, with speeds of 5.5-6 km/h and with water at different depths: baseline (BL, exercise in water treadmill without water), and with the water at level of the metacarpophalangeal/metatarsophalangeal (MCPJ; MTPJ), tarsal (TJ) and stifle joints (SJ). Hematology and blood lactate (LA) were measured before and after each session, and heart rate (HR) was monitored with a pulsometer. Horses also wore a portable gait analyzer, consisting of three orthogonal accelerometers, fixed at the sternum and at the sacrum, and the following parameters were measured: stride length (SL), stride frequency (SF), regularity (REG, measurement of the acceleration pattern similarity of successive strides in a period of time), symmetry (SYM, measurement of the similarities between left and right acceleration patterns), dorsoventral displacement (DVD, displacement of the gravity center in a dorsoventral direction), dorsoventral DVP, propulsion PP, mediolateral MLP and total power TP, representing the amount of acceleration and deceleration along the dorsoventral, longitudinal and lateral axes and the sum of the three powers respectively. Significant changes in hematological parameters and in blood LA concentrations before and after each exercise session were not found. Blood LA concentrations after BL, MCPJ, TJ and SJ sessions reached mean values of 1.03±0.41, 1.05±0.28, 1.05±0.52 and 1.11±0.22 mmol/l respectively. HR increased significantly in the four sessions, with significant differences after exercise between BL (66.13±8.92 bpm), TJ (81.89±10.67 bpm) and SJ (81.13±14.68 bpm) levels. Further, significant differences were also found between exercise at the level of MCPJ (71.09±8.90 bpm) and TJ. With the accelerometer fixed on the sternum, increased SL, DVD, DVP and TP together with a decreased SF was found with the water at the level of TJ, compared to the other water depths. With the accelerometer fixed on the sacrum, significant changes were not observed in SF between the different water depths. The highest values for SL, DVD, DVP, PP, MLP and TP were found with the water at the level of the TJ. Our results reveal that exercise in WT, nevertheless of the water depth (at least until stifle joint), represents a mild aerobic exercise, with low HR and blood LA under the aerobic threshold of 2 mmol/l. However, this type of exercise leads to profound changes in locomotion, mainly in stride length and frequency and in muscle power, with the highest values with water depths at the tarsal and stifle joints. Consequently, the low cardiovascular and metabolic demands are expected to be of interest for the recovery of
{"title":"Exercise in water in the horse: physiological adaptations and potential benefits for training and rehabilitation","authors":"A. Muñoza","doi":"10.13130/2283-3927/10268","DOIUrl":"https://doi.org/10.13130/2283-3927/10268","url":null,"abstract":"Recently, a scientific interest in exercise in water in equine athletes, both from training and rehabilitation purposes has been developed. Before an extended use of this practice in equine exercise physiology and medicine, the physiological adaptations to this type of exercise need to be elucidated. Six horses were subjected to sessions of 40 minutes in water treadmill, with speeds of 5.5-6 km/h and with water at different depths: baseline (BL, exercise in water treadmill without water), and with the water at level of the metacarpophalangeal/metatarsophalangeal (MCPJ; MTPJ), tarsal (TJ) and stifle joints (SJ). Hematology and blood lactate (LA) were measured before and after each session, and heart rate (HR) was monitored with a pulsometer. Horses also wore a portable gait analyzer, consisting of three orthogonal accelerometers, fixed at the sternum and at the sacrum, and the following parameters were measured: stride length (SL), stride frequency (SF), regularity (REG, measurement of the acceleration pattern similarity of successive strides in a period of time), symmetry (SYM, measurement of the similarities between left and right acceleration patterns), dorsoventral displacement (DVD, displacement of the gravity center in a dorsoventral direction), dorsoventral DVP, propulsion PP, mediolateral MLP and total power TP, representing the amount of acceleration and deceleration along the dorsoventral, longitudinal and lateral axes and the sum of the three powers respectively. Significant changes in hematological parameters and in blood LA concentrations before and after each exercise session were not found. Blood LA concentrations after BL, MCPJ, TJ and SJ sessions reached mean values of 1.03±0.41, 1.05±0.28, 1.05±0.52 and 1.11±0.22 mmol/l respectively. HR increased significantly in the four sessions, with significant differences after exercise between BL (66.13±8.92 bpm), TJ (81.89±10.67 bpm) and SJ (81.13±14.68 bpm) levels. Further, significant differences were also found between exercise at the level of MCPJ (71.09±8.90 bpm) and TJ. With the accelerometer fixed on the sternum, increased SL, DVD, DVP and TP together with a decreased SF was found with the water at the level of TJ, compared to the other water depths. With the accelerometer fixed on the sacrum, significant changes were not observed in SF between the different water depths. The highest values for SL, DVD, DVP, PP, MLP and TP were found with the water at the level of the TJ. Our results reveal that exercise in WT, nevertheless of the water depth (at least until stifle joint), represents a mild aerobic exercise, with low HR and blood LA under the aerobic threshold of 2 mmol/l. However, this type of exercise leads to profound changes in locomotion, mainly in stride length and frequency and in muscle power, with the highest values with water depths at the tarsal and stifle joints. Consequently, the low cardiovascular and metabolic demands are expected to be of interest for the recovery of","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86791975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10041
S. Giorgi, M. Comí, M. Ghiringhelli, V. Bontempo
In the search for the reduction of antibiotics in farm animals, a concept was developed based on studies with medium-chain fatty acid with 6-12 atoms (MCFA). In particular, they have been shown to exhibit against Gram+ bacteria in piglets at relatively high concentrations. However they can be hardly used as such because of their repellent odour and taste and for their rapid absorption in upper gastrointestinal tract. These problems could be overcome by the generation of monoacylglycerol, but esterification is usually carried out on a silica base, which reduces the concentration of FA, therefore limiting the antibacterial effects. Our hypothesis is that the saponification with calcium salts might positively affect their concentration in the GIT. The aim of the present study was to examine the effects of Laurate calcium soap (C12-Ca soap) on growth performance and health of post-weaning piglets. At weaning, 192 crossbreed Topics piglets were assigned to 3 experimental groups consisting of 16 pens (4 pigs/pen each): CTR (negative control), T1 (basal diet plus Amoxycillin at 400 mg/kg), and T2 (basal diet plus C12-Ca soap at 1 kg/ton). Gain and feed consumption did not differ among groups. Feed efficiency was higher in T1 (0,61) and T2 (0,58) than CTR (0,51) (P<0,01). Mortality was 0 in T1, and reduced in T2 (4,7%) compared to CTR (10,9%). These preliminary results suggest that saponification of MCFA may be a valuable alternative to in-feed antibiotics, used for growth promotion, and even for enhancing health in post-weaning piglets.
{"title":"The saponification of lauric acid with calcium soaps as an alternative to in-feed antibiotics in post-weaning piglets","authors":"S. Giorgi, M. Comí, M. Ghiringhelli, V. Bontempo","doi":"10.13130/2283-3927/10041","DOIUrl":"https://doi.org/10.13130/2283-3927/10041","url":null,"abstract":"In the search for the reduction of antibiotics in farm animals, a concept was developed based on studies with medium-chain fatty acid with 6-12 atoms (MCFA). In particular, they have been shown to exhibit against Gram+ bacteria in piglets at relatively high concentrations. However they can be hardly used as such because of their repellent odour and taste and for their rapid absorption in upper gastrointestinal tract. These problems could be overcome by the generation of monoacylglycerol, but esterification is usually carried out on a silica base, which reduces the concentration of FA, therefore limiting the antibacterial effects. Our hypothesis is that the saponification with calcium salts might positively affect their concentration in the GIT. The aim of the present study was to examine the effects of Laurate calcium soap (C12-Ca soap) on growth performance and health of post-weaning piglets. At weaning, 192 crossbreed Topics piglets were assigned to 3 experimental groups consisting of 16 pens (4 pigs/pen each): CTR (negative control), T1 (basal diet plus Amoxycillin at 400 mg/kg), and T2 (basal diet plus C12-Ca soap at 1 kg/ton). Gain and feed consumption did not differ among groups. Feed efficiency was higher in T1 (0,61) and T2 (0,58) than CTR (0,51) (P<0,01). Mortality was 0 in T1, and reduced in T2 (4,7%) compared to CTR (10,9%). These preliminary results suggest that saponification of MCFA may be a valuable alternative to in-feed antibiotics, used for growth promotion, and even for enhancing health in post-weaning piglets.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81026523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-06-12DOI: 10.13130/2283-3927/10002
E. Manzoni, S. Arcuri, T. Brevini, F. Gandolfi
In the last years, many papers highlighted the possibility to use epigenetic modifiers to directly interact with the epigenetic signature of an adult mature cell (Pennarossa et al., 2013; Chandrakantan et al., 2016). In particular, the molecule 5-azacytidine (5-aza-CR), which is able to interfere with DNA methylation, through both a direct and an indirect effect (Manzoni et al., 2016), can be used to remove the epigenetic ‘blocks’ responsible for tissue specification and to facilitate cell transition to a different lineage. In parallel, recent evidence has also shown that epigenetic conversion is influenced by the 3D rearrangement and by the mechanical properties of the cellular microenvironment (Pennarossa et al., 2017). In the experiments here presented, we investigated the effect of a selected 3D culture system on the conversion process. We used INS-eGFP porcine fibroblasts, that express enhanced green fluorescent protein (eGFP) under the control of insulin gene promoter, as experimental model, and wild-type pig fibroblasts, as control. Both cell types , were plated either on plastic or on 1kPa polyacrylamide (PAA) gel, that mimics the stiffness of pancreatic tissue in vivo . Cells were erased with 5-aza-CR for 18h and exposed to specific differentiation stimuli for 36 days (Pennarossa et al., 2014). The use of INS-eGFP fibroblasts allowed real-time monitoring of cells progressing towards the pancreatic phenotype. Morphological analysis and pancreatic marker expression were checked for the entire length of the experiment. PAA gels encouraged the induction of islet-like structures, suggesting that the of tridimensional clusters may be a crucial aspect of pancreatic differentiation in vitro . Moreover, the use of an adequate substrate accelerated cell differentiation process and anticipated insulin secretion ability. The results obtained demonstrated the direct implication of the yes-associated protein/transcriptional co-activator with PDZ-binding motif (YAP/TAZ) mechanotransduction-mediated pathway, indicating that mechanical cues exert a key role in pancreatic phenotype definition.
在过去的几年中，许多论文强调了使用表观遗传修饰剂直接与成年成熟细胞的表观遗传特征相互作用的可能性(Pennarossa et al.， 2013;Chandrakantan et al.， 2016)。特别是分子5-氮杂胞苷(5-aza-CR)，它能够通过直接和间接的影响干扰DNA甲基化(Manzoni等人，2016)，可以用来去除负责组织规范的表观遗传“块”，并促进细胞向不同谱系的转变。与此同时，最近的证据也表明，表观遗传转化受到3D重排和细胞微环境的机械特性的影响(Pennarossa et al.， 2017)。在这里提出的实验中，我们研究了选定的3D培养系统对转化过程的影响。我们以胰岛素基因启动子调控下表达增强型绿色荧光蛋白(eGFP)的INS-eGFP猪成纤维细胞为实验模型，以野生型猪成纤维细胞为对照。这两种类型的细胞要么被镀在塑料上，要么被镀在1kPa的聚丙烯酰胺(PAA)凝胶上，模仿体内胰腺组织的硬度。细胞用5-aza-CR擦除18h，并暴露于特定分化刺激下36天(Pennarossa et al.， 2014)。使用INS-eGFP成纤维细胞可以实时监测细胞向胰腺表型发展。形态学分析和胰腺标志物表达在整个实验期间进行检查。PAA凝胶促进胰岛样结构的诱导，表明三维簇的形成可能是体外胰腺分化的一个关键方面。此外，使用适当的底物加速细胞分化过程和预期的胰岛素分泌能力。结果表明，yes相关蛋白/转录共激活因子与pdz结合基序(YAP/TAZ)机械转导介导途径直接相关，表明机械提示在胰腺表型定义中发挥关键作用。
{"title":"Matrix stiffness boosts pancreatic differentiation via the YAP/TAZ mechanotransduction mediated pathway","authors":"E. Manzoni, S. Arcuri, T. Brevini, F. Gandolfi","doi":"10.13130/2283-3927/10002","DOIUrl":"https://doi.org/10.13130/2283-3927/10002","url":null,"abstract":"In the last years, many papers highlighted the possibility to use epigenetic modifiers to directly interact with the epigenetic signature of an adult mature cell (Pennarossa et al., 2013; Chandrakantan et al., 2016). In particular, the molecule 5-azacytidine (5-aza-CR), which is able to interfere with DNA methylation, through both a direct and an indirect effect (Manzoni et al., 2016), can be used to remove the epigenetic ‘blocks’ responsible for tissue specification and to facilitate cell transition to a different lineage. In parallel, recent evidence has also shown that epigenetic conversion is influenced by the 3D rearrangement and by the mechanical properties of the cellular microenvironment (Pennarossa et al., 2017). In the experiments here presented, we investigated the effect of a selected 3D culture system on the conversion process. We used INS-eGFP porcine fibroblasts, that express enhanced green fluorescent protein (eGFP) under the control of insulin gene promoter, as experimental model, and wild-type pig fibroblasts, as control. Both cell types , were plated either on plastic or on 1kPa polyacrylamide (PAA) gel, that mimics the stiffness of pancreatic tissue in vivo . Cells were erased with 5-aza-CR for 18h and exposed to specific differentiation stimuli for 36 days (Pennarossa et al., 2014). The use of INS-eGFP fibroblasts allowed real-time monitoring of cells progressing towards the pancreatic phenotype. Morphological analysis and pancreatic marker expression were checked for the entire length of the experiment. PAA gels encouraged the induction of islet-like structures, suggesting that the of tridimensional clusters may be a crucial aspect of pancreatic differentiation in vitro . Moreover, the use of an adequate substrate accelerated cell differentiation process and anticipated insulin secretion ability. The results obtained demonstrated the direct implication of the yes-associated protein/transcriptional co-activator with PDZ-binding motif (YAP/TAZ) mechanotransduction-mediated pathway, indicating that mechanical cues exert a key role in pancreatic phenotype definition.","PeriodicalId":14105,"journal":{"name":"International Journal of Health, Animal science and Food safety","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77146881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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