Ingestion of Bacillus anthracis spores causes gastrointestinal (GI) anthrax. Humoral immune responses, particularly immunoglobulin A (IgA)-secreting B-1 cells, play a critical role in the clearance of GI pathogens. Here, we investigated whether B. anthracis impacts the function of colonic B-1 cells to establish active infection. GI anthrax led to significant inhibition of immunoglobulins (eg, IgA) and increased expression of program death 1 on B-1 cells. Furthermore, infection also diminished type 2 innate lymphoid cells (ILC2) and their ability to enhance differentiation and immunoglobulin production by secreting interleukin 5 (IL-5). Such B-1-cell and ILC2 dysfunction is potentially due to cleavage of p38 and Erk1/2 mitogen-activated protein kinases in these cells. Conversely, mice that survived infection generated neutralizing antibodies via the formation of robust germinal center B cells in Peyer's patches and had restored B-1-cell and ILC2 function. These data may provide additional insight for designing efficacious vaccines and therapeutics against this deadly pathogen.

Canine osteosarcoma is an extremely malignant bone tumor that often arises in the bones of the limbs. It is a highly metastatic disease distinguished by proliferative bone lesions and a tendency for pulmonary metastasis. Overexpression of proliferative proteins are associated with bad prognosis in human osteosarcoma. Here, we tested the expression of the different proliferative proteins (p53, p16, vimentin, and mdm2) in nine archival samples with canine osteosarcoma. Paraffin-embedded tissue sections were confirmed by histopathology and stained by immunohistochemistry for p53, p16, vimentin and mdm2. Positive expression of these proteins was evaluated as the ratio of positive cancer cells and the intensity staining was assessed in several areas. Histopathologically, 95% of samples were grade II and III. All high-grade osteosarcomas were particularly cellular. The cancer cells were generally large spindle-shaped and large nucleus with distribution of osteoid between the cancer cells. Immunohistochemical detection of p53, p16, vimentin and mdm2 was 89%, 56%, 78%, and 89% of samples respectively. The staining intensity for p53, p16, vimentin and mdm2 was particularly nuclear in 81%, 66%, 78%, and 79% of the cancer cells respectively. Our present work suggests that p53, p16, vimentin, and mdm2 were detected in grade III canine osteosarcomas samples. In addition, these proliferative markers are the significant biomarkers in canine osteosarcomas and can be used as a predictor for diagnostic and prognostic value and allowing cancer differentiation. This primary data supports that both canine and human osteosarcomas share same molecular characters which are approved by expression of proliferative genes.

Pheochromocytomas (PCs) are tumors arising from the chromaffin cells of the adrenal glands and are the most common tumors of the adrenal medulla in animals. In people, these are highly correlated to inherited gene mutations in the succinate dehydrogenase (SDH) pathway; however, to date, little work has been done on the genetic basis of these tumors in animals. In humans, immunohistochemistry has proven valuable as a screening technique for SDH mutations. Human PCs that lack succinate dehydrogenase B (SDHB) immunoreactivity have a high rate of mutation in the SDH family of genes, while human PCs lacking succinate dehydrogenase A (SDHA) immunoreactivity have mutations in the SDHA gene. To determine if these results are similar for dogs, we performed SDHA and SDHB immunohistochemistry on 35 canine formalin-fixed, paraffin-embedded (FFPE) PCs. Interestingly, there was a loss of immunoreactivity for both SDHA and SDHB in four samples (11%), suggesting a mutation in SDHx including SDHA. An additional 25 (71%) lacked immunoreactivity for SDHB, while retaining SDHA immunoreactivity. These data suggest that 29 out of the 35 (82%) may have an SDH family mutation other than SDHA. Further work is needed to determine if canine SDH immunohistochemistry on PCs correlates to genetic mutations that are similar to human PCs.

To investigate the incidental histopathologic findings in clinically normal dogs and determine the immunohistochemical expression of several tissue-specific differentiation markers. The University of Mosul's animal research ethical committee authorized this study (UoM.Dent/A.DM.L.1/22). Seven samples were obtained from the veterinary hospital in cooperation with the College of Dentistry. Dogs with two detectable testicles were considered intact. Dogs with a history of clinical signs of prostatic or lower urinary tract disease were dismissed from the study. Sections were cut at 5µm, and then hematoxylin and eosin staining procedures were applied. Immunohistochemistry was performed following the manufacturer's protocol for each antibody. Typical prostatic glandular features lined by simple columnar epithelial cells with apical intracytoplasmic eosinophilic granules were detected, which represent a major basic protein. Additionally, despite a lack of gross lesions, 13 types of microscopic lesions were identified. Infiltration by lymphocytes and plasma cells was the most frequent, in 85.71% (6/7) of samples. Atrophied glands and congested blood vessels were noticed in 71.42% (5/7). Intraluminal glandular hyperplasia, epithelial vacuolar degeneration, and increased fibrous tissue were detected in 57.14% (4/7). Increased interstitial stroma was found in 42.85% (3/7). Polypoid projections, hyaline cast deposition, cystic alveoli, and epithelial lining deterioration play a significant role in the inflammatory process, prostate concretions and capsule thickening were rarer abnormalities. P63, cytokeratin 8, and high molecular weight cytokeratin were used to target normal prostate gland cells in clinically normal mixed-breed dogs. We also investigated the expression of BCL -2 and P53, which have been recorded in human prostatic cancer. P63 was detected in all samples and CK8 was detected in 7/8 samples. No samples had immunoreactivity for BCL-2 and P53. Prostate samples from normal dogs show a spectrum of lesions, with a majority of samples having lymphoplasmacytic inflammation. IHC found weak to strong P63 immunoreactivity and strong CK8 immunoreactivity, with no reactivity for P53, BCL-2, and HMWC. All statistical work, both descriptive and inferential, was accomplished in JMP Pro 16.1. Analyzing with a chi-squared test. By showing similar lesions to those seen in humans, even in normal cases, these results further suggest that dogs may be a suitable model for human prostatic illness. Also, caution is warranted while looking for small neoplasms, as these lesions may hide small neoplasm foci.

Fixation inhibits autolysis and is an essential step in disease. Our work examined how neutral buffer NBF (NBF), zinc formalin (ZF), and Bouin's fluid (BF) affected tissue specific CD4, CD68, and Pax8 anti-genetic determinants. Fifty-one sections were analyzed for each fixative marker. One hundred fifty-three portions were scored 0, 1, 2, and 3. The comparison between scores in the detection of CD4 epitopes reflects that BF fluid was significant compared to NBF and ZF at a level of p< 0.05, whereas NBF comes after ZF. Surprisingly, ZF has no significant scores in the context of CD4 detection. On the other hand, CD68 detection at the score one level revealed that NBF and ZF showed significant CD68 anti-genetic determinants, whereas none of the CD68 epitopes were significantly determined in BF fluid. Eventually, both NBF and BF produced an excellent determination of Pax8 epitopes in contrast to ZF. However, there is no significant difference between them. Regarding scores three comparisons in all fixatives, BF fluid showed high-quality CD4 marker expression in contrast to NBF and ZF; however, neither CD68 nor Pax8 showed any significance in this context. We conclude that NBF showed an excellent quality result in detecting all epitopes. However, BF fluid was supreme in detecting anti-genetic determinants of CD4 and Pax8 epitopes in score 3. Additionally, ZF could distinguish CD68 and Pax8 but failed to detect CD4. Conversely, no significant differences were recorded between both B and NBF.

DNA extraction was successfully performed on all samples with variable concentrations. Fifteen liver samples were subjected to the extraction technique, with five liver samples utilized from each fixative, this comparison study aimed to assess DNA integrity across different fixatives. DNA was successfully extracted from all samples regardless of their varying quantities. Each sample yielded at least one motif with a length of 100 base pairs. All 15 samples produced 100 bp amplicon for the IRBP gene. All 15 samples generated a 200 bp amplicon from the NBF, ZF, and BF. The success proportion dropped with growing amplicon length, by which 66.6% (10/15) of 300 bp amplicons were successfully produced from NBF and ZF, showing significant differences at P<0.05 than BF, while no other amplicon length obtained 0% for 400, 500, and 750 bp respectively, on the other hand, frozen samples worked as a control were able to produce all amplicon length from 100bp to 750bp. Only samples from NBF and ZF constantly yielded more extensive amplicon ranges than BF. BF was unable to keep intact DNA fragments of more than 200 bp. Our study concluded that different fixatives could produce at least 100 bp and 200 bp. However, ZF and NBF are more dependable than BF in producing 300 bp for downstream analysis for further next-generation techniques. Furthermore, NBF and ZF are supreme in producing better-quality DNA fragments than BF.