Inhibin-α, E-cadherin, calretinin and Ki-67 antigen in the immunohistochemical evaluation of canine and human testicular neoplasms

Introduction. The steady increase of dogs with diagnosed testicular neoplasms observed in recent years prompted us to carry out immunohistochemical (IHC) studies for their better characterization. The aim of the study was to analyze most common canine testicular neoplasms (seminomas, Leydig cell and Sertoli cell tumors) with selected IHC markers and to compare the expressions of these proteins in corresponding canine and human testicular tumors. Material and methods. Studies were carried out on testicular canine tumors: 40 cases of seminoma, 40 cases of Leydig cell tumor and 40 cases of Sertoli cell tumor. Moreover, 15 cases of human seminomas and 5 cases of human Leydig cell tumors were also analyzed. Immunohistochemistry was performed on paraffin sections by standard technique using monoclonal anti-human antibodies against E-cadherin, inhibin-α, calretinin and Ki-67. The slides were subjected to computer-aided image analysis and the intensity of the immunoreactivity was assessed by a semi-quantitative scoring system. Results. Due to the very low prevalence of the Sertoli cell-derived tumors in the human population, we were able to examine the markers’ expression only in the canine gonadal tumors. We revealed that, apart from E-cadherin in Leydig cell tumors and calretinin in seminomas, the expression of all the analyzed markers in canine and human testicular tumors was similar. E.g. there was no immunoexpression of inhibin-α in 75% of canine and 100% of human cases of seminoma. The immunoreactivity of Ki-67 was intense in 40% of canine and 60% of human seminomas. Immunoexpression of inhibin-α in Leydig cell tumor was intense in 70% of canine and 100% of human cases, respectively. Also the immunoreactivity of calretinin was intense in 75% of cases of canine and 100% of human Leydig cell tumors. In 50% of canine and 40% of human Leydig cell tumors, the immunoexpression of Ki-67 was weak. Conclusions. The applied anti-human monoclonal antibodies against common antigens and markers of human testicular neoplasms could be routinely used for the immunohistochemical evaluation of canine testicular tumors.

Equine Stomach Development in the Fetal Period: An Anatomical, Topographical, and Morphometric Study

Studies of equine stomach prenatal development are very rare, and descriptions usually focus on the processes taking place in the embryonic period. Only general information about gastric organogenesis in the fetal period is available in embryology textbooks on domestic mammals. The material for our study included twenty half-breed horse fetuses divided into three age groups on the basis of known fetal age (verified using the CRL method). Our study consists of the topographical, morphological, and morphometrical description of stomach development between the 4th and 11th months of gestation. Even though the skeletotopy, syntopy, and holotopy of the stomach in the fetal period seems to be relatively unchanged, the organ shape and the proportions between its anatomical parts differed in fetuses from the three age groups. The achieved results were statistically elaborated to estimate the dynamics of the stomach shape. This can be described as changing from medium-wide to wide and from slightly bent to sharply bent. A nonlinear correlation of all metric values with CRL in all age groups was observed. A positive allometric growth rate of different intensity was seen in all metric parameters. All the values increased as the fetal period progressed. Only the parietal surface growth rate gradually changed from strongly positive allometric in the first age group to strongly negative allometric in the third age group. A difference between the non-glandular and glandular mucosa of the stomach was visible in the first group. Development of a well-distinguishable plicated edge margin began in the second age group together with gastric pits and gastric areas. The third age group showed a well-developed gastric groove and angular incisura

Effect of selected nonsteroidal anti-inflammatory drugs on the viability of canine osteosarcoma cells of the D-17 line: in vitro studies

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used in veterinary medicine. They are used in pain control and in anti-inflammatory and antipyretic therapies. Some NSAIDs, e.g piroxicam, also have a documented anticancer effect. The objective of this study was to evaluate which of the commonly used NSAIDs (etodolac, flunixin, tolfenamic acid, carprofen, and ketoprofen) are cytotoxic to the D-17 cell line of canine osteosarcoma

Histological and Electrophoretic Analysis of Carpathian barbel (Barbus carpathicus, Cyprinidae) Skin and Mucus in Environmental Context

Fish frequently serve as bioindicators of aquatic environments during their ecological evaluation. Carpathian barbel (Barbus carpathicus, Cyprinidae) is a species common to rivers and lakes of Eurasia and Africa. Seasons of the year can influence its skin morphology and mucus composition. The clinical status of the animal depends on the above-mentioned factors. The aim of this study was a histological, histometrical and electrophoretical analysis of periodical changes in barbel common integument. The accessible material was investigated in histological, cytological and electrophoretic analysis using hematoxylin-eosin staining, histometric morphometry, gel electrophoresis and cytological methods. The results demonstrated significant differences in the investigated parameters for spring–summer and autumn–winter periods. Both skin epithelium morphology (epithelium thickness, number of cell layers, melanophores and mucous cell existence) and mucus composition (proteins, immune system cells, keratinocytes and mucocytes) showed significant differences between investigated seasons. These morphological and physiological changes were more pronounced in the dorsal than ventral regions of common integument. The differences in the physical characteristics of mucus and the histological structure of the skin cannot only serve as a source of useful information about an evaluated ecosystem, but can be also related to additional factors, e.g., microbiological and chemical water contamination

Synergistic Antitumor Interaction of Risedronate Sodium and Standard Anticancer Agents in Canine (D-17) and Human Osteosarcoma (U-2 OS) Cell Lines

The study discusses in vitro cytotoxicity of a combination of cytostatic drugs (doxorubicin, cisplatin, carboplatin, etoposide) and risedronate sodium against canine and human osteosarcoma (D-17 and U-2 OS). Standard protocols were used for the preparation of cell cultures and evaluation of their viability and apoptosis. MTT assay assessed the culture viability and EC50, while the apoptotic effect of the drugs was checked with a TUNEL assay. Doxorubicin alone showed the strongest cytotoxicity against D-17 (0.056 ± 0.019 µg/mL) and U-2 OS (0.051 ± 0.003 µg/mL), while the lowest cytotoxicity was observed for carboplatin (D-17, 6.45 ± 0.2 µg/mL and U2-OS, 27.5 ± 2.3 µg/mL). Risedronate sodium at 100, 10 and 1 µg/mL lowered viability in OS cell lines by 53.38 ± 1.46 and 49.56 ± 0.7%, 97.08 ± 3.32 and 74.92 ± 4.01%, and 102.67 ± 3.56 and 94.56 ± 3.52%, respectively. In all analyzed drug combinations, risedronate sodium significantly (* p osteosarcoma cell lines. The decrease in cell viability caused by the studied compound combinations was weaker in canine than in human cell cultures. A combination of doxorubicin (all concentrations), cisplatin (1 µg/mL) and etoposide (1 µg/mL) with 100 µg/mL of risedronate sodium significantly improved the cytotoxicity of the drugs against canine and human osteosarcoma. Administration of carboplatin (1 µg/mL) and risedronate sodium (100 µg/mL), compared to carboplatin per se, produced no significant differences in cytotoxicity against the D-17 cell culture but significantly enhanced cytotoxicity in the U-2 OS line. The strongest apoptosis in both lines was detected for 0.01 µg/mL doxorubicin combined with 100 µg/mL risedronate sodium or 1 µg/mL cisplatin and 100 µg/mL risedronate sodium. In all combinations, the tested compounds revealed a synergistic mechanism of action

Dogs in the Wroclaw Stronghold, 2nd Half of the 10th–1st Half of the 13th Century (Lower Silesia, Poland)—An Zooarchaeological Overview

This article pertains to the issue of early medieval dogs (10th–mid-13th century) from the territory of Poland and Central Europe. The study is based on dog remains from the Wroclaw Cathedral Island (Ostrów Tumski), one of the most important administrative centres of early medieval Poland, the capital of a secular principality and the seat of diocese authorities. The main morphological and functional types of dogs living in Wroclaw and other parts of Poland were characterized on that basis. It has been concluded that the roles and perceptions of dogs were very ambiguous. On the one hand, they were hunting companionship for the elite and were considered a symbol of devotion and loyalty. On the other hand, dogs symbolised disgrace. In everyday life, these animals were sometimes abused, their skin was sometimes tanned and their bones modified into tools, and in exceptional cases, dogs were even eaten

The iron age dogs from alaybeyi höyük, eastern anatolia

To date, little is known about the biological and cultural status of Iron Age dogs in Anatolia. Here, we present a zooarchaeological study of an assemblage of 143 Iron Age dog bones, including two dog skeletons, unearthed from the 2016 and 2017 salvage excavations at Alaybeyi Höyük, Eastern Anatolia. At least eight adults and one juvenile individual, along with a large number of miscellaneous specimens, were identified. The morphological status of the Alaybeyi dogs were primarily compared to previously published Iron Age dogs from Yoncatepe in Eastern Anatolia, and with the average mean of 18 modern dog breeds. Unlike in other Eastern Anatolian Iron Age sites, butcher marks were observed in some specimens, indicating at least occasional cynophagy at the site. Noticeable pathologies were found in about 5% of the sample, particularly pathologies of the oral cavity and dentitions, suggesting that some of the dogs at Alaybeyi Höyük might have been undernourished, had to live on solid food, and probably injured by humans. The results of this study reflect both the morphological and biological status of Alaybeyi dogs, as well as the Alaybeyi people’s attitudes toward dogs, adding vital information to the very limited archaeological knowledge of dogs in Anatolia

In Vitro Studies on the Influence of Meloxicam on Cytotoxic Activity Induced by Risedronate Sodium in Canine (D-17) and Human (U-2 OS) Osteosarcoma Cell Lines

The study describes the cytotoxic effect against human and canine osteosarcoma (U-2 OS and D-17) cell lines induced by risedronate sodium and meloxicam per se and in combination. Both cell lines were prepared according to standard procedures for cell cultures studies. The cell viability was estimated in both cell lines treated with chosen concentrations of risedronate sodium and meloxicam. The apoptosis assessment was carried out using TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay. EC50 values, computed for risedronate sodium and meloxicam cytotoxicity, showed comparable effects against the canine OS cell line in similar concentration of both drugs. In case of human OS, the stronger cytotoxic effect of risedronate sodium was proved. The EC50 values for meloxicam in both cell lines were, statistically, significantly different (* p < 0.05). Moreover, the cytotoxic effect of a combined administration of meloxicam and risedronate sodium in doses 100 µg/mL, compared with the negative control showed statistically significant differences. The human OS cell line was more resistant to both compounds than the canine OS cell line. The apoptotic effect in canine and human osteosarcoma triggered by risedronate sodium and meloxicam was statistically significant (p < 0.05). The cytotoxic effect induced with 100 µg/mL of risedronate sodium proved statistically significant differences between both tested cell lines compared to negative control. The results obtained with 10 and 100 µg/mL of meloxicam were not statistically significant. The study showed the synergic mechanism of action of risedronate sodium and meloxicam, but the concentrations used in vitro will not be possible to achieve in in vivo. Therefore, our results serve as basis only to design future studies on the tissue level