Histo Hepatic Alterations in Commercially Important Fish (Pseudoplatystoma corruscans Spix & Agassiz, 1829) Exposed to a Glyphosate-Based Herbicide

The presence of herbicides in the aquatic environment can cause different effects at all levels of biological organization. This study aimed to evaluate the hepatic alterations of the herbicide Roundup WG® on juvenile Pseudoplatystoma corruscans exposed to three different concentrations of this chemical compound: 0.25g/1000L, 0.50g/1000L, and 0.75g/1000L, plus control treatment (0.00g/1000L). The experiment lasted sixty days and, in the end, liver fragments were collected for further histological processing, using the hematoxylin-eosin (HE) technique. Only the control group showed a statistically significant increase in body mass and total length during the experiment. Blood glucose also showed no difference among the sample groups. The lesions found in the liver considered severe were hemorrhage, vacuolization, and hypertrophy of hepatocytes and the presence of free melanomacrophages, recorded in the groups exposed to 0.50 and 0.75g/1000L. The results obtained in the present study indicate that the herbicide Roundup WG® can promote liver alterations in Pseudoplatystoma corruscans.

Considerations regarding liver current pathology

This work presents reasoning about liver pathology. Liver pathology is an area of interest due to the growing number of medical cases worldwide. The factors associated with the appearance of this pathology are those that relate to individual lifestyle. Liver pathology studies are performed up to the biomolecular level, involving cytochrome P450 2E1 (CYP2E1). With a normal liver, subject to aging and the aforementioned risk factors, there are changes in liver structure, which often can lead to changes arising from nonalcoholic hepatic steatosis (NASH). Under certain circumstances this degenerates into chronic hepatitis, which often goes undiagnosed and this can lead to liver cirrhosis. Due to gradual changes in the liver, medical interventions are often delayed. Importantly, structural analysis of microscopically processed liver fragments will reveal changes in all structural elements of the liver, from lipid loading in hepatic steatosis to inflammatory, necrotic, destructive, fibrotic changes in liver cirrhosis, including somewhat similar changes in types of chronic hepatitis. The presented images and the described structural changes in the liver are an important criterion for liver damage, including preclinical pathology.

Mitochondria Load Degree in Hepatocytes of the Classical Hepatic Lobules in Chinchilla (Chinchilla lanigera)

Hepatocytes represent the majority of the liver cell population and are arranged in the form of cords placed in intimate contact with the sinusoidal capillaries. The functional complexity corroborated with the intensity of the activity of hepatocytes requires large amounts of energy. The organelles involved in the production of chemical energy used in the activity of hepatocytes are the mitochondria. The purpose of this study was to verify the mitochondrial load of hepatocytes in all areas of the classical hepatic lobules, in order to indirectly assess the intensity of hepatocyte activity in each area. Materials and Methods Five fresh corpses of chinchilla (Chinchilla lanigera) from an independent breeder from Bistrița-Năsăud county were used. Liver fragments were harvested and fixated in Kolster’s solution for 24 hours, stained with Heidenhain ferric hematoxylin, and assessed using Olympus BX41 microscope. Fixation with Kolster's solution and the staining with Heidenhain's iron hematoxylin clearly shows the hepatocytic mitochondria in shades from gray to black. The liver lobules displayed an uneven distribution of mitochondria depending on the area. In zone 1 of the classical hepatic lobule, the degree of loading of hepatocytes with mitochondria is larger than in zone 2 and much larger than in zone 3. Morphological features of the hepatocytes, including the number and distribution of mitochondria in the hepatic lobules, should improve the understanding of the physiology and pathology of the liver.

Clinical, imaginological and pathological aspects of umbilical panvasculitis in calves: case report

The diagnosis of umbilical infections in neonates can be obtained from clinical signs, but the intracavitary involvement of structures and associated complications can be underestimated, compromising the establishment of adequate therapeutic approaches or prognosis. This case report presents the clinical, imaging, pathological and microbiological aspects of an umbilical infection in calves. Physical examination of the animal identified apathy, low body score, increased volume in the umbilical region and joints. The abdominal palpation identified firm structures in topography of the arteries and umbilical vein. Imaging examinations of the abdomen and joints were performed. Multiple, hyperechogenic focal structures have been identified in the liver, as well as cylindrical and firm structures in topography of the arteries and umbilical vein. In the joints, osteolytic changes, periosteal reactions, subchondral sclerosis and formation of osteophytes were seen. Umbilical panvasculitis triggered arthritis and an infectious process in the liver, the case being assessed as having an unfavorable prognosis and the animal being referred for euthanasia. At necropsy, multifocal abscesses were observed in the pleura, ribs, omentum, spleen and liver. There was granulomatous exudate in the urinary vesicle. The affected joints presented thickening of the joint capsule with the presence of exudate. In the microbiological analysis of liver fragments, urinary vesicle content and joint exudate, Proteus mirabilis with resistance to antimicrobials was identified. Imaging studies collaborated with the establishment of the prognosis and conduct adopted, and must, whenever possible, be included in the clinical examination. In case of death, necropsy allows a correct association of clinical signs and imaging findings.

Effect of supercritical carbon dioxide on the in vivo biocompatible and resorptive properties of tissue-specific scaffolds from decellularized pig liver fragments

The creation of bioengineered tissue/organ equivalents is closely related to the development of biodegradable, highly porous 3D scaffolds, which to some extent provide the microenvironment necessary to maintain the viability of the cellular component. According to many researchers, the most interesting are tissue-specific matrices that can selectively support the adhesion, proliferation and differentiation of tissue cells of those organs from which they are obtained by decellularization. It was shown that during intramuscular implantation in rats of decellularized pig liver fragments (DLFp), independent of the method of detergent residues removing (96 hours of washing in phosphate-salt buffer (PBS) or combined: 24 hours in PBS and 8 hours with supercritical CO2 (sc-CO2), the samples meet the requirements for medical devices in terms of local and general toxic effects. Thus, the use of sc-CO2 made it possible to reduce the duration of the technology for producing biocompatible tissue-specific matrices based on DLFp by 3 times. Moreover, when using sc-CO2 at the stage of washing the DLFp matrix, a “mild reaction” of the tissue to the sample is observed during 2 months of intramuscular implantation of the matrix in rats with its complete resorption after 3 months of the experiment. Under the same conditions, the duration of a similar local action of DLFp washed in the PBS on the tissue is 3 months with degradation of 63% of the matrix of the sample size.

Spontaneous poisoning by Cestrum intermedium in dairy cattle

ABSTRACT: Acute hepatotoxicity caused by plants poisoning is responsible for economic losses in farm animals in Brazil. Reports of Cestrum intermedium natural poisoning in cattle are not commonly described in Rio Grande do Sul (RS). This study aimed to document an outbreak of spontaneous C. intermedium poisoning in dairy cattle in the Central-Eastern Mesoregion of RS. Three nine-month-old Holstein and Jersey heifers were affected after they were placed in a small paddock with shortage forage. In this area, specimens of C. intermedium Sendtn with signs of consumption were observed. Morbidity and lethality rates were 100% and clinical courses ranged from 9 to 12 hours. At post mortem examination of the three heifers, there was predominance of acute liver lesions. The liver was moderately enlarged and on the cut surface there was a marked accentuation of the lobular pattern and hemorrhage. Inside the rumen, partially digested C. intermedium Sendtn leaves were observed. The histological aspects of the liver were mostly centrilobular coagulative necrosis and hemorrhage, frequently extended to the midzonal region. The immunohistochemistry technique was performed, in which the polyclonal antibody caspase 3 was used in liver fragments. Moderate to marked immunolabeling was observed in the cytoplasm and nucleus of hepatocytes, predominantly on the periphery of areas of hepatic necrosis indicating cell apoptosis. The diagnosis of C. intermedium Sendtn poisoning in dairy cattle in this study was based on epidemiological, clinical and anatomopathological findings. Since the C. intermedium poisoning is uncommon in dairy cattle, we are describing it for the first time in the Central-Eastern Mesoregion of RS, and represents a differential diagnosis of other acute toxic liver diseases in cattle.

Experimental approaches to creating a tissue-specific matrix for a bioartificial liver

Shortage of donor organs for liver transplantation in the treatment of end-stage liver disease dictates the need to develop alternative methods that include technologies on tissue engineering and regenerative medicine. Objective: to study the ability of a tissue-specific matrix from decellularized human liver fragments (DHLF) to maintain adhesion and proliferation of human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) and HepG2 under static conditions and in a flow-through bioreactor. Materials and methods. Treatment with surfactants (SAS) – sodium dodecyl sulfate, Triton X-100 – followed by exposure to DNase was used for decellularization of human liver fragments (no more than 8 mm3). Biochemical screening included the determination of DNA quantity in the test samples. Efficiency of surfactant washing was assessed by the cytotoxicity of the matrix in the NIH 3T3 fibroblast culture. Viability and metabolic activity of cells were assessed via vital staining with a complex of fluorescent dyes LIVE/DEAD ® and PrestoBlue™ (Invitrogen, USA). Morphological examination of the liver cell-engineered constructs was carried out through histological staining and scanning electron microscopy with lanthanide contrast. Results. It was shown that the liver decellularization method used allows to obtain a biocompatible matrix with a residual DNA quantity <1%, which is capable of maintaining adhesion and proliferation of hAT-MSCs and HepG2. On day 7 of cultivation in the bioreactor, there was formation of a single conglomerate of the DHLF matrix with numerous groups of viable cells with a high nuclear-cytoplasmic ratio. The urea content in the culture medium is 1.5 ± 0.1 mmol/L, exceeding that of samples obtained under static conditions. This indicates the metabolic activity of HepG2 in the composition of the obtained culture systems. It was shown that constant flow of the culture medium in the perfusion bioreactor increased the proliferative activity of HepG2 and allowed to provide a more uniform colonization by matrix cells in comparison with static cultivation conditions. Conclusion. The conditions for uniform colonization of DHLFs in a flow-through bioreactor with cell cultures were established. The ability of the matrix to maintain adhesion and proliferation of hADSCs and HepG2 for 11 days indicates that it could be used in liver tissue engineering.

Identification, Expression, and Functions of the Somatostatin Gene Family in Spotted Scat (Scatophagus argus)

Somatostatins (SSTs) are a family of proteins consisting of structurally diverse polypeptides that play important roles in the growth regulation in vertebrates. In the present study, four somatostatin genes (SST1, SST3, SST5, and SST6) were identified and characterized in the spotted scat (Scatophagus argus). The open reading frames (ORFs) of SST1, SST3, SST5, and SST6 cDNA consist of 372, 384, 321, and 333 bp, respectively, and encode proteins of 123, 127, 106, and 110 amino acids, respectively. Amino acid sequence alignments indicated that all SST genes contained conserved somatostatin signature motifs. Real-time PCR analysis showed that the SST genes were expressed in a tissue specific manner. When liver fragments were cultured in vitro with synthetic peptides (SST1, SST2, or SST6 at 1 μM or 10 μM) for 3 h or 6 h, the expression of insulin-like growth factor 1 and 2 (Igf-1 and Igf-2) in the liver decreased significantly. Treatment with SST5 had no significant effect on Igf-1 and Igf-2 gene expression. This study provides an enhanced understanding of the gene structure and expression patterns of the SST gene family in S. argus. Furthermore, this study provides a foundation for future exploration into the role of SST genes in growth and development.

The beginning of the program of multiorgan postmortem donation and transplantation in the Republic of Sakha (Yakutia)

The article is devoted to the main aspects of organization of multiorgan donation from donors with brain death in the Republic of Sakha (Yakutia). Transplant services to the population of this region have been in place since 2001, and until 2016 only kidney transplants or liver fragments from a living related donor have been performed. The presence of an effective postmortem donation program will allow to meet the high demand of this region for high-tech assistance by organ transplantation.

Technology features of decellularization of human liver fragments as tissue-specific fine-grained matrix for cell-engineering liver construction

One of the problems when you create a bioengineered liver, as an alternative to transplantation of the donor liver in the treatment of end-stage liver failure, is the matrix, able to temporarily perform the functions of the natural extracellular matrix (ECM) and provide the necessary conditions to maintain the viability of the liver cells. The main disadvantage of resorbable biopolymer matrices is the absence of tissue specifi c properties and the impossibility of reproducing the unique structure of the ECM of the liver.Aim:to develop technology for decellularization of liver tissue fragments, saving the structural properties of native ECM of the liver.Materials and methods.The decellularization of mechanically grinded human liver fragments was carried out in three changes of buffer solution (pH = 7.4) containing 0.1% sodium dodecyl sulfate and increasing the concentration of Triton X100 (1%, 2% and 3%, respectively). During technology development were investigated the effects of duration, conditions (static, dynamic, rotary system, magnetic stirrer) washing and methods of liver tissue grinding on the completeness of removal of cellular elements and detritus preserving the the liver ECM structure. Slices of decellularized liver tissue samples were stained by hematoxylin and eosin, and Masson method for the detection of connective-tissue elements.Results.Histological analysis methods showed that the best from the point of view of effi ciency of decellularization and the safety of the structure of own human liver ECM, is a mode of washing of liver fragments for three days at room temperature in static conditions, accompanied by stirring by a magnetic stirrer for 2–3 times a day for one hour. Longer time or a large multiplicity of mixing mode is accompanied by increased risk of liver tissue damage. On the basis of the experimental results obtained the algorithm of preliminary study of donor human liver designed to optimize the process of obtaining decellularization fragments of liver tissue was elaborated.Conclusion.It was elaborated the algorithm and technology of obtaining of decellularized liver tissue fragments from the human donor liver which saved the structural properties of native ECM of the liver and complete removal of cellular elements and detritus.