COMPLETED PROJECTS
Hepatotoxicity and immunophenotypic analysis of lymphocytes
We performed histological technique and pathological evaluation of liver samples from mice that received experimental T cell-based immunotherapy. The hepatotoxicity assessment of obtained slides was performed in accordance with international criteria, including the International Harmonization of Nomenclature and Diagnostic Criteria (INHAND), issued by The Global Editorial and Steering Committee (GESC). We performed histological technique designed to isolate genetic material form material embedded in paraffin blocks and allows to use molecular biology methods as well as laser microdissection.
Furthermore, we conduct flow cytometry analysis of 30 peripheral blood samples from mouse of same study by BD FACS Canto II flow cytometer (BD Bioscience) and the FlowJo software analysis program. In the gating strategy, lymphocytes (small, non-granular cells - P1), single cells (Singlets) and live cells (Viable) were analyzed with control of an unstained sample (Unstain) and a double isotype control (Cocktail and APC). The following cell labeling was included: CD3+CD4+ (T helper cells; Th), CD3+CD8+ (cytotoxic T cells; Tc) and CD3-NKp46+ (NK cells).
Inflammatory bowel diseases and gastrointestinal lymphomas
The aim of this study is to analyze the potential impact of gut microbial metabolites on lymphoproliferative diseases development in animals and comparison of the data with human research outcomes. The research includes histopathological classification of inflammatory bowel diseases and gastrointestinal tract lymphomas in dogs and cats, along with evaluation of cell immunophenotype, proliferation parameters and markers of lymphoid line differentiation and maturation.
Advanced digital pathology system and the 3DHISTECH histological scanner allows us to perform morphometrics of morphological classification criteria of gastrointestinal lymphomas along with optical density analysis of immunohistochemical staining. Moreover, we have developed a protocol for isolation of ribonucleic acid (RNA) from paraffin blocks which is next analyzed for expression of the gut microbial metabolites receptors genes as AHR, FFAR2/GPR43 i FFAR3/GPR41. Our team is also performing immunohistochemical staining for examined receptors with optical density analysis as well as determining the subcellular location and level of analyzed proteins.
European bison keratoconjunctivitis
ALAB bioscience laboratory, as a part of larger project, conducted histopathological and morphometric analysis of European bison’s eyeballs with keratoconjunctivitis. Considering European bison species characteristics, we carried out morphometric analysis of corneal layer and designed histopathological severity scale which allows pathologist to make semi-quantitative assessment of keratoconjunctivitis lesions.
What is more, our team developed and performed a histological technique which enable additional molecular diagnostics methods of the examined eyeballs. We isolated genetic material from paraffin blocks and optimize the PCR reaction conditions what allowed us to look for potential infectious factors that could cause conjunctivitis and keratitis in bison.
Furthermore, we have developed a high-throughput screening profile based on Biomark HD technique for monitoring infectious diseases of European bison’s, this endangered and protected species in Poland.
Protective effect of plant origin drug in cadmium toxicity
ALAB bioscience laboratory were part of a protective effectiveness project of plant origin drug in nephrotoxicity induced by the administration of cadmium as a mouse model of environmental exposure to humans. We performed histological technique and standard hematoxylin and eosin (HE) staining as well as Masson's trichrome staining.
Moreover, we designed histopathological severity scale of nephrotoxicity in accordance with international criteria, including the International Harmonization of Nomenclature and Diagnostic Criteria (INHAND), issued by The Global Editorial and Steering Committee (GESC). Our team determined the relationship between effect and animal sex, exposure time and cadmium concentration. We have also prepared a full photographic documentation of histological slides, including comparison micrographs.
Acute toxicity of graphene
ALAB bioscience pathologists’ team performed a toxicopathological assessment in conformity with OECD protocol „Guideline for the Testing of Chemicals No. 420 (2001): Acute oral toxicity – fixed dose procedure” i procedurą UE B.1.BIS: „Acute oral toxicity – fixed dose procedure”. The study consists of dosing rats intraperitoneally with single, high dose of graphene oxide nanoflakes coated with metalized platinum nanoparticles. Our team examined the histological slides of kidneys, liver, heart, lungs, and spleen and assessed the acute in accordance with international criteria, including the International Harmonization of Nomenclature and Diagnostic Criteria (INHAND), issued by The Global Editorial and Steering Committee (GESC).
The study was realized within the framework of cooperation with the WULS at SGGW.
Determining the role of the protein product of the Zc3h12a gene - the Mcpip1 protein - in liver function
In collaboration with the Department of General Biochemistry of the Faculty of Biochemistry, Biophysics and Biotechnology at the ALAB bioscience laboratory, we conducted a series of histopathological, biochemical and hematological studies on mice subjected to selective deletion of the Zc3h12a gene in hepatocytes. The goal of the study was to determine the role of the protein product of the Zc3h12a gene - the Mcpip1 protein - in liver function. A team of our pathologists, together with biochemists at WBBiB UJ, interpreted the image, the obtained pathological, molecular and biochemical changes, to show features of Primary Biliary Cholongitis (PBC). In subsequent studies, our team performed histopathological classification of human livers for the diagnosis of non-alcoholic fatty liver disease (NAFLD, or Nonalcoholic Fatty Liver Disease) and evaluation of fibrosis of this organ.