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The GM2-gangliosidoses are neurological diseases causing premature death, thus developing effective treatment protocols is urgent. GM2-gangliosidoses result from deficiency of a lysosomal enzyme β-hexosaminidase (Hex) and subsequent accumulation of GM2 gangliosides. Genetic changes in HEXA, encoding the Hex α subunit, or HEXB, encoding the Hex β subunit, causes Tay–Sachs disease and Sandhoff disease, respectively. Previous studies have showed that a modified human Hex µ subunit (HEXM) can treat both Tay–Sachs and Sandhoff diseases by forming a homodimer to degrade GM2 gangliosides. To this end, we applied this HEXM subunit in our PS813 gene editing system to treat neonatal Sandhoff mice. Through AAV delivery of the CRISPR system, a promoterless HEXM cDNA will be integrated into the albumin safe harbor locus, and lysosomal enzyme will be expressed and secreted from edited hepatocytes. 4 months after the i.v. of AAV vectors, plasma MUGS and MUG activities reached up to 144- and 17-fold of wild-type levels (n = 10, p

Deficiencies in the lysosomal hydrolase β-galactosidase (β-gal) lead to two distinct diseases: the skeletal disease Morquio syndrome type B, and the neurodegenerative disease GM1-gangliosidosis. Utilizing CRISPR-Cas9 genome editing, the mouse β-gal encoding gene, Glb1, was targeted to generate both models of β-gal deficiency in a single experiment. For Morquio syndrome type B, the common human missense mutation W273L (position 274 in mice) was introduced into the Glb1 gene (Glb1W274L), while for GM1-gangliosidosis, a 20 bp mutation was generated to remove the catalytic nucleophile of β-gal (β-gal−/−). Glb1W274L mice showed a significant reduction in β-gal enzyme activity (8.4–13.3% of wildtype), but displayed no marked phenotype after one year. In contrast, β-gal−/− mice were devoid of β-gal enzyme activity (≤1% of wildtype), resulting in ganglioside accumulation and severe cellular vacuolation throughout the central nervous system (CNS). β-gal−/− mice also displayed severe neuromotor and neurocognitive dysfunction, and as the disease progressed, the mice became emaciated and succumbed to the disease by 10 months of age. Overall, in addition to generating a novel murine model that phenotypically resembles GM1-gangliosidosis, the first model of β-galactosidase deficiency with residual enzyme activity has been developed.

In the present study, we aimed to assess the toxicity of artemisinin on the haematological system and its effect on the productive performance of broiler chickens. Eighteen-day-old chickens were randomly divided into four groups of 30 chickens (three replicates of 10 broilers/group): control group and three experimental groups: ART5 - diet with 5 ppm of artemisinin; ART50 - diet with 50 ppm of artemisinin; and ART500 - diet with 500 ppm of artemisinin. Artemisinin enhanced the productive performances of broiler chickens at the lowest concentration (5 ppm), but at the highest concentration (500 ppm), it negatively affected weight gain and the feed conversion ratio. The performance characteristics of the chickens whose diets were supplemented with 50 ppm artemisinin were similar to those of the control group. Additionally, 5 ppm artemisinin in feed did not significantly affect the haematological parameters of the chickens, but 50 and 500 ppm artemisinin induced a gradual decline of the total leukocytes, lymphopenia, monocytosis, and eosinopenia, and the highest concentration caused anaemia. Artemisinin at a low concentration could be used as a feed additive in the poultry industry to improve organic broiler production performance with no serious side effects.

Primary spindle cell neoplasms of the gastrointestinal system show considerable morphologic overlaps. Proper histological classification of these neoplasms using only the classical hematoxylin-eosin (HE) stained slides is often difficult and frequently a source of diagnosis error. Thus, the immunohistochemistry is an essential tool in the proper diagnosis and classification of such tumors.In this study we tested and standardised an immunohistochemical protocol used for the differentiation of primary intestinal sarcomas (nonangiogenic, nonlymphogenic intestinal sarcomas) in dogs. The paper also aimed to critically review the state of art in immunohistochemistry of intestinal spindle cell neoplasms currently used in dogs.Seven cases of primary intestinal spindle cell neoplasms were diagnosed and classified using a 6 antibody panel immunohistochemistry. Indirect immunohistochemistry staining for S100 protein, desmin, I±-SMA, CD34, c-Kit and GFAP was conducted using a Leica Bond-Max auto-immunostainer.Based on the immunoreactivity, five stromal tumours were classified as leiomyosarcomas, one fibrosarcoma and one an undifferentiated sarcoma.In most cases of intestinal sarcomas, the proper diagnosis requests the use of immunohistochemistry. The retrospective analysis of these tumours proved that at least in one case, the HE stain was not sufficient for the proper diagnosis and classification of the tumour.

Lipid-rich invasive human breast cancer is a rare enigmatic entity among special types of infiltrating duct carcinoma. Our paper reports a lipid-rich mammary carcinoma in a female cat with the gross, microscopic and immunohistochemical description of the tumor. A 13-year-old intact adult female, mixed-breed cat was presented by the owner to the Laboratory of Pathologic Anatomy from the Faculty of Veterinary Medicine Cluj-Napoca, Romania. A complete necropsy examination was performed in our laboratory. The tissue samples were collected and processed by paraffin technique for further histological, histochemical, immunofluorescence and immunohistochemical examination. During the necropsy examination, a subcutaneous mass was discovered on the chest. Several variably sized, well-demarcated neoplasms were noted in the right axillary lymph node, right thoracic wall, pleura, lungs, liver, spleen and kidney. Histologically, the cells frequently formed tubuloacinar structures. The morphology of the described tumor showed features of a poorly differentiated mammary carcinoma. Numerous tumoral cells were large and polygonal, with abundant cytoplasm that showed foam-like cytoplasm. The tumoral cells contained either multiple small or large and solitary vacuoles that pushed the nucleus to the periphery of the cell. Intracytoplasmic vacuoles of the neoplastic cells were positive for Oil-Red-O and negatively with Periodic Acid–Schiff. As for immunofluorescence/immunohistochemistry, nonvacuolated and vacuolated neoplastic cells were positive for cytokeratin and negative for vimentin. Histochemical and immunohistochemical analysis support a diagnosis of lipid-rich mammary carcinoma. This is the second reliable record of a lipid-rich mammary carcinoma in female cat and the first one with internal metastases.