Use of Cold Atmospheric Plasma in the Treatment of Squamous Cell Carcinoma: in vitro Effects and Clinical Application in Feline Tumors: A Pilot Study.

Cold atmospheric plasma (CAP) has shown promising results against squamous cell carcinoma (SCC) in both in vivo and in vitro assays, mainly in humans and mice. Its applicability for treatment of feline tumors, however, remains unknown. This study aimed to evaluate the anticancer effects of CAP on a head and neck squamous cell carcinoma (HNSCC) cell lineage and against a clinical case of cutaneous SCC in a cat. Control and treatment groups employing the HNSCC cell line (SCC-25) were used, the latter exposed to CAP for 60 seconds, 90 seconds, or 120 seconds. The cells were subjected to the MTT assay nitric oxidation assay and thermographic in vitro analyses. The clinical application was performed in one cat with cutaneous SCC (3 sites). The lesions were treated and evaluated by thermographic, histopathological, and immunohistochemical examinations (caspase-3 and TNF-alpha). Treatment of the SCC-25 cells for 90 seconds and 120 seconds resulted in a significant nitrite concentration increase. Decreased cell viability was observed after 24 hours and 48 hours, regardless of exposure time. However, the cell viability reduction observed at 72 hours was significant only in the 120 seconds treatment. In vitro, the temperature decreased for all treatment times, while the plasma induced a slight increase in mean temperature (0.7°C) in the in vivo assay. Two of the 3 clinical tumors responded to the treatment: one with a complete response and the other, partial, while the third (lower lip SCC) remained stable. Both remaining tumors displayed apoptotic areas and increased expression of caspase-3 and TNF-alpha. Adverse effects were mild and limited to erythema and crusting. The CAP exhibited an in vitro anticancer effect on the HNSCC cell line, demonstrated by a dose-dependent cell viability reduction. In vivo, the therapy appears safe and effective against feline cutaneous SCC. The treatment did not result in a clinical response for 1 of 3 lesions (proliferative lower lip tumor), however, a biological effect was still demonstrated by the higher expression of apoptosis indicators.

Feline injection site sarcoma: immunohistochemical characteristics.

OBJECTIVES: Feline injection site sarcoma (FISS) is a rapid growing locally aggressive tumor with a low metastatic rate. Its histologic features are clearly defined, but there are few studies regarding its immunohistochemical characteristics. The present study investigated the immunohistochemical characteristics of 21 cases of FISS. METHODS: FISSs from 12 male and nine female cats, 20 mixed-breed and one Siamese, were included in the study. After histopathological diagnosis, additional histologic sections were immunostained for vimentin, cytokeratin, desmin, S100 protein, viral feline leukemia virus (FeLV) particles, cyclooxygenase 2 (COX-2) and c-KIT. Positive and negative controls were adopted accordingly. Immunostainings were classified as positive or negative according to the number of positive cells from a total of 1000 cells per tumor section. RESULTS: Histopathologic diagnosis of the tumors revealed 18 (85.7%) fibrosarcomas and three (14.3%) other sarcomas; four fibrosarcomas (22.2%) were grade III, five (27.8%) were grade II and nine (50.0%) were grade I. Two sarcomas were grade III and one was grade II. Seventeen (81%) tumors were negative for desmin. All samples were positive for vimentin. Twenty tumors (95.2%) were positive for S-100 protein. Positivity for c-KIT was observed in four (19%) samples; COX-2 was positive in 13 (61.9%) and FeLV viral particles were positive in nine (42.9%) FISSs. CONCLUSIONS AND RELEVANCE: Immunohistochemical findings of FISSs revealed positive immunostainings for desmin, vimentin, S-100 protein, c-KIT, COX-2 and FeLV viral particles.

Imunolocalização de receptores de leptina no ovário de preás (Galea spixii Wagler, 1831) / Immunolocalization of leptin receptor of Spix's Yellow-toothed Cavy (Galea spixii Wagler, 1831) ovary

A leptina, uma citocina produzida pelas células adiposas, é alvo da comunidade científica por acreditarem que ela apresente impacto sobre a reprodução dos animais promovendo a puberdade, foliculogênese e oogênese, ciclo estral e auxiliando na fecundação. A compreensão dos mecanismos que controlam a atividade reprodutiva de preás (Galea spixii) possui papel relevante para a preservação da espécie. Desta forma, o presente trabalho propôs analisar a imunolocalização dos receptores de leptina (Ob-R) no ovário de preás. Coletaram-se os ovários de 20 fêmeas adultas, não prenhes e saudáveis. As amostras foram fixadas em paraformaldeído a 4% em tampão fosfato, incluídas em parafina e seccionadas para a realização de imunohistoquímica (IHC). As secções foram fotomicrografadas e avaliadas quanto à intensidade da reação. Observou-se forte imunorreação no oócito e nas células da teca, moderada nas células do estroma ovariano e nas células luteínicas grandes e fracamente coradas nas células da granulosa, endoteliais, perivasculares e células luteínicas pequenas. Quando comparado a expressão de receptores ao longo do desenvolvimento folicular foi observado que o oócito e as células da teca se mantiveram com expressão na mesma intensidade. Entretanto, as células da granulosa apresentaram forte marcação nos estádios pré-antrais enquanto que nos folículos antrais apresentou fraca intensidade. Concluímos que em ovários de Galea spixii existe a presença de Ob-R nas principais estruturas do ovário sugerindo que este hormônio desempenhe papel fundamental na reprodução desta espécie.

Leptin, a cytokine produced by adipose cells, is the target of the scientific community for believing that it has an impact on the reproduction of the animals promoting puberty, folliculogenesis and oogenesis, estrous cycle and aiding in fertilization. The understanding of the mechanisms controlling the reproductive activity of Spix's Yellow-toothed Cavy (Galea spixii) plays a relevant role in the preservation of the species. Thus, the present study proposed to analyze the immunolocalization of leptin receptors (Ob-R) in the ovary of cavies. Ovaries from 20 adult, non-pregnant, healthy females were collected. The samples were fixed in 4% phosphate buffered paraformaldehyde, embedded in paraffin and sectioned for immunohistochemistry. The sections were photomicrographs and intensity of the reaction was measured. Strong immunoreaction was observed in oocyte and theca cells, moderate in ovarian stromal cells and large luteal cells and weak stained in granulosa, endothelial, perivascular and small luteal cells. When compared to receptor expression along follicular development it was observed that the oocyte and the theca cells remained with expression at the same intensity. However, the granulosa cells presented strong stained in the preantral stages, whereas in the antral follicles it presented low intensity. We conclude that in the ovaries of Galea spixii there is the presence of Ob-R in the main structures of the ovary sugesting that this hormone plays a fundamental role in the reproduction of this species.

Predictive values of aspartate aminotransferase and gamma-glutamyl transferase for the hepatic accumulation of copper in cattle and buffalo.

Ten cattle and 10 buffalo were divided into 2 groups (control [n = 8] and experimental [n = 12] that received daily administration of copper. Three hepatic biopsies and blood samples were performed on days 0, 45, and 105. The concentration of hepatic copper was determined by spectrophotometric atomic absorption, and the activities of aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) were analyzed. Regression analyses were done to verify the possible existing relationship between enzymatic activity and concentration of hepatic copper. Sensitivity, specificity, accuracy, and positive and negative predictive values were determined. The serum activities of AST and GGT had coefficients of determination that were excellent predictive indicators of hepatic copper accumulation in cattle, while only GGT serum activity was predictive of hepatic copper accumulation in buffalo. Elevated serum GGT activity may be indicative of increased concentrations of hepatic copper even in cattle and buffalo that appear to be clinically healthy. Thus, prophylactic measures can be implemented to prevent the onset of a hemolytic crisis that is characteristic of copper intoxication.