Immunolocalization of angiotensin-(1-7), angiotensin II and angiotensin-converting enzyme 2 in goat ovary

There is increasing evidence as to the participation of the ovarian renin-angiotensin system in important reproductive processes. The inhibition of the angiotensin-converting enzyme (ACE) caused an increase in the rate of ovulation and pregnancy in the artificial insemination protocol has fixed time (TFIA). This study aimed to investigate the presence and location of Ang II, Ang- (1-7) and ACE2 in goat ovaries and the possibility of the involvement of these peptides in previous results. Ten ovaries from goats were collected in a slaughterhouse, washed in buffered PBS, perfused with protease inhibitor solution and processed for immunohistochemistry protocol. The search for peptides was performed using the avidin–biotin–peroxidase method. A strong immunoreactivity for Ang II in theca cells of antral follicles and corpus luteum was observed. Antral follicles (theca cells), corpus luteum and oocyte cytoplasm in early antral follicles exhibited strong immunoreactivity for Ang-(1-7). There was strong immunoreactivity for ACE2 in the cytoplasm of luteal cells and theca cells of antral follicles. In this study, for the first time, the presence and location of Ang II, Ang-(1-7) and ACE2 are reported in goat ovary, suggesting that there is participation in follicular development, oocyte maturation and corpus luteum development.

Characteristics of a Novel ATP2B3 K416_F418delinsN Mutation in a Classical Aldosterone-Producing Adenoma

In patients with primary aldosteronism (PA), the prevalence of ATP2B3 mutation is rare. The aim of this study is to report a novel ATP2B3 mutation in a PA patient. Based on our tissue bank of aldosterone-producing adenomas (APA), we identified a novel somatic ATP2B3 K416_F418delinsN mutation. The affected individual was a 53 year-old man with a 4 year history of hypertension. Computed tomography (CT) showed bilateral adrenal masses of 1.6 (left) and 0.5 cm (right) in size. An adrenal venous sampling (AVS) showed a lateralization index (LI) of 2.2 and a contralateral suppression index (CLS) of 0.12; indicating left functional predominance. After a left unilateral adrenalectomy, he achieved partial biochemical and hypertension–remission. This classical adenoma harbored a novel ATP2B3 K416_F418delinsN somatic mutation, which is a deletion from nucleotides 1248 to 1253. The translated amino acid sequence from 416 to 418, reading as lysine-phenylalanine-phenylalanine, was deleted; however, an asparagine was inserted due to merging of residual nucleotide sequences. The CYP11B2 immunohistochemistry staining demonstrated strong immunoreactivity in this classical adenoma. The ATP2B3 K416_F418delinsN mutation is a functional mutation in APA, since HAC15 cells, a human adrenal cell line, transfected with the mutant gene showed increased CYP11B2 expression and aldosterone production.

Metabolic Disorders/Obesity Is a Primary Risk Factor in Hidradenitis Suppurativa: An Immunohistochemical Real-World Approach

<b><i>Background:</i></b> Hidradenitis suppurativa (HS) is an inflammatory, potentially scarring disease of the hair follicle, affecting the apocrine gland-bearing skin areas. The major comorbid disorders associated with the occurrence or the aggravation of the disease are obesity and smoking. Numerous efforts to dissociate these factors led to controversial results. <b><i>Objectives:</i></b> To assess the importance of metabolic disorders/obesity, smoking/environmental toxins, and inflammation in HS by utilizing the differential expression of major relevant protein markers in lesional skin of obese/smoking versus non-obese/non-smoking HS patients. <b><i>Methods:</i></b> Lesional skin specimens deriving from two groups of HS patients (BMI &#x3e;30 and smokers, <i>n</i> = 12 vs. BMI &#x3c;30 and non-smokers, <i>n</i> = 10) were stained with antibodies raised against irisin, PPARγ, and IGF-1R, which correlate with metabolic disorders/obesity, EGFR and AhR, associated with smoking, and IL-17, IL-17R, and S100A8, as markers of inflammation. <b><i>Results:</i></b> Metabolic disorders/obesity-related markers exhibited marked differential expression between the two groups, while smoking-associated markers a limited one. IL-17R expression was stronger in obese/smokers, and S100A8 staining exhibited intense strong immunoreactivity in both groups without significant difference. <b><i>Conclusions:</i></b> The notion that obesity plays a role in HS development appears to be supported by the prominent regulation of the associated lesional biomarkers. Tobacco smoking might contribute less to HS than previously suspected.

Molecular Characterization and Expression Analysis of Annexin B3 and B38 as Secretory Proteins in Echinococcus Granulosus

Background: Cystic Echinococcosis is a parasitic zoonotic disease, which poses a threat to public health and animal husbandry, and causes significant economic losses. Annexin is a kind of phospholipid binding protein with calcium ion binding activity, which has many functions.Methods: Two annexin protein family genes (EgAnxB3 and EgAnxB38) were cloned and molecular characterized using bioinformatic analysis. The immunoreactivity of rEgAnxB3 and rEgAnxB38 was investigated using western blotting. The distribution and transcript levels of EgAnxB3 and EgAnxB38 in different developmental stages of Echinococcus granulosus were analyzed by immunofluorescence localization and quantitative real-time reverse transcription PCR, and their secretory characteristics were analyzed preliminary. The phospholipid-binding activities of rEgAnxB3 and rEgAnxB38 were also tested.Results: EgAnxB3 and EgAnxB38 are conserved and contain calcium binding sites. Both rEgAnxB3 and rEgAnxB38 could be specifically recognized by the serum samples from E. granulosus-infected sheep, which have strong immunoreactivity. EgAnxB3 and EgAnxB38 were distributed in all stages of E. granulosus and had high transcript levels in the 28-day strobilated worms. They were found in liver tissues near the cysts. In addition, rEgAnxB3 had Ca2+-dependent phospholipid-binding properties.Conclusions: EgAnxB3 and EgAnxB38 contained calcium-binding sites, and rEgAnxB3 had Ca2+-dependent phospholipid-binding properties. rEgAnxB3 and rEgAnxB38 had strong immunoreactivity. EgAnxB3 and EgAnxB38 were transcribed in PSCs and worms. They were expressed in all stages of E. granulosus, and distributed in the liver tissues near the hydatid cyst, indicating that EgAnxB3 and EgAnxB38 are secreted proteins that play an crucial role in the development of E. granulosus.

Molecular characterization of CNS paragangliomas identifies cauda equina paragangliomas as a distinct tumor entity

Paragangliomas/pheochromocytomas are rare neuroendocrine tumors that arise from the adrenal gland or ganglia at various sites throughout the body. They display a remarkable diversity of driver alterations and are associated with germline mutations in up to 40% of the cases. Comprehensive molecular profiling of abdomino-thoracic paragangliomas revealed four molecularly defined and clinically relevant subtypes. Paragangliomas of the cauda equina region are considered to belong to one of the defined molecular subtypes, but a systematic molecular analysis has not yet been performed. In this study, we analyzed genome-wide DNA methylation profiles of 57 cauda equina paragangliomas and show that these tumors are epigenetically distinct from non-spinal paragangliomas and other tumors. In contrast to paragangliomas of other sites, chromosomal imbalances are widely lacking in cauda equina paragangliomas. Furthermore, RNA and DNA exome sequencing revealed that frequent genetic alterations found in non-spinal paragangliomas—including the prognostically relevant SDH mutations—are absent in cauda equina paragangliomas. Histologically, cauda equina paragangliomas show frequently gangliocytic differentiation and strong immunoreactivity to pan-cytokeratin and cytokeratin 18, which is not common in paragangliomas of other sites. None of our cases had a familial paraganglioma syndrome. Tumors rarely recurred (9%) or presented with multiple lesions within the spinal compartment (7%), but did not metastasize outside the CNS. In summary, we show that cauda equina paragangliomas represent a distinct, sporadic tumor entity defined by a unique clinical and morpho-molecular profile.

Comprehensive genomic analysis contrasting primary colorectal cancer and matched liver metastases: a retrospective study

(Background)Recent studies have revealed that colorectal cancer (CRC) displays intratumor genetic heterogeneity, and the cancer microenvironment play an important role in the proliferation, invasion, and metastasis of CRC.(Methods)We performed genomic analysis on 22 paired patients with primary CRC and synchronous colorectal liver metastasis (CRLM) using whole-exome sequencing, cancer gene panels, and microarray gene expression profiling. In addition, immunohistochemical analysis was used to confirm the protein expression of genes identified as highly expressed in CRLM by DNA microarray analysis.(Results)A total of 22 paired patients were enrolled in this study, including 13 (59.1%) colon and 9 (40.9%) rectal cancer patients. All patients had synchronous liver metastasis and did not have any other extrahepatic metastases. We identified 11 probes (10 genes) that were highly expressed in CRLM compared to CRC, from 36022 probes obtained from primary CRC, CRLM, and normal liver tissues, using gene expression analysis with DNA microarray. Of the 11 probes (10 genes), 5 genes classified as encoding “matricellular proteins” (osteopontin, periostin, thrombospondin-2, MGP, and GPNMB) were selected for immunohistochemical analysis.Osteopontin was strongly expressed in CRLM (6 of 22 cases: 27.3%) but not in CRC (0 of 22: 0%; p=0.02). Periostin also showed strong immunoreactivity in CRLM (17 of 22: 68.2%) compared to 7 of 22 (31.8%) in CRC (p=0.006). Thrombospondin-2 showed strong immunoreactivity both in CRC and CRLM (54.5% in CRC, 45.5% in CRLM; p=0.55). GPNMB and MGP were rarely positive for both CRC and CRLM. A comparison of immunoreactive positive factors for these 5 genes revealed the complexities of gene expression in CRLM. Of the cases, 16 (72.7%) CRC revealed zero or only one positive immunoreactivity. On the other hand, CRLM revealed more frequent multiple immunoreactivity; 16 cases (72.7%) shared 2 or more factors, which was statistically more frequent in CRLM than in CRC (p=0.007).(Conclusions)This study revealed the genomic heterogeneity between paired primary CRC and CRLM in terms of cancer cell microenvironment. This finding will lead to new diagnostic and therapeutic targets in the era of genome-guided personalized cancer treatment.(Trial registration)This study was retrospectively registered.

Primary malignant melanoma arising from ruptured ovarian mature cystic teratoma with elevated serum CA 19–9: a case report and review of literature

Background Ovarian mature cystic teratomas comprise tissues derived from all three germ layers. In rare incidences, malignant tumors may arise from ovarian mature cystic teratoma, which occurs in 0.2–1.8% of cases. A variety of tumors can arise within mature cystic teratoma, among which malignant melanoma is exceedingly rare. Case presentation A 42-year-old woman presented with abdominal pain. Transvaginal ultrasonography showed mixed echogenic cystic masses in both ovaries. Her serum cancer antigen (CA19–9) level was elevated at 29,770 U/ml. Surgical excision was performed. Histologic examination showed infiltrating nests of pleomorphic cells with prominent nucleoli and black pigments in the background of a mature cystic teratoma. These pleomorphic cells showed strong immunoreactivity for Melan-A and HMB-45. The patient was re-evaluated and the possibility of a melanoma at any other site was ruled out. Based on these findings, we concluded that the malignant melanoma originated from the ovarian mature cystic teratoma. Conclusion We report a rare case of primary malignant melanoma derived from an ovarian mature cystic teratoma.

Morphologic and Immunohistochemical Characterization of Spontaneous Lymphoma/Leukemia in NSG Mice

The NOD.Cg- Prkdcscid Il2rgtm1Wjl/SzJ strain (NOD scid gamma, NSG) is a severely immunodeficient inbred laboratory mouse used for preclinical studies because it is amenable to engraftment with human cells. Combining scid and Il2rgnull mutations results in severe immunodeficiency by impairing the maturation, survival, and functionality of interleukin 2–dependent immune cells, including T, B, and natural killer lymphocytes. While NSG mice are reportedly resistant to developing spontaneous lymphomas/leukemias, there are reports of hematopoietic cancers developing. In this study, we characterized the immunophenotype of spontaneous lymphoma/leukemia in 12 NSG mice (20 to 38 weeks old). The mice had a combination of grossly enlarged thymus, spleen, or lymph nodes and variable histologic involvement of the bone marrow and other tissues. All 12 lymphomas were diffusely CD3, TDT, and CD4 positive, and 11 of 12 were also positive for CD8, which together was consistent with precursor T-cell lymphoblastic lymphoma/leukemia (pre-T-LBL). A subset of NSG tissues from all mice and neoplastic lymphocytes from 8 of 12 cases had strong immunoreactivity for retroviral p30 core protein, suggesting an association with a viral infection. These data highlight that NSG mice may develop T-cell lymphoma at low frequency, necessitating the recognition of this spontaneously arising disease when interpreting studies.

Incidence, Gross Morphology, Histopathology and Immunohistochemistry of Canine Mammary Tumors

The aim of present investigation was to study the gross morphology and incidence of canine mammary tumors (CMTs) based on age, sex, breed, reproductive status and location along with histopathological classification and immunohistochemical characteristics. A total of 56 CMTs samples were collected from 49 cases of dogs. Gross morphology was studied in 26 cases of canine mammary tumors. For histopathological classification, samples were fixed in 10% formalin, embedded in paraffin and sections (5 μm thick) obtained from each was stained with HandE stain. Immuno-histochemistry was carried out by using p63 antibody to confirm the histopathological types of CMTs. Malignant tumors and benign tumors were mostly observed in older dogs. Among 9 breeds affected, the highest incidence was recorded in a German shepherd. Caudal abdominal pair was most commonly affected. Most of the cases were observed in intact female dogs, except for one male dog. The tumors were oval and round in shape with 30–2000 g weight, soft to hard in consistency and grayish white cut surface. Out of 56 CMTs, the highest incidence was found of malignant neoplasms (36/56, 64.28%), followed by benign neoplasms (10/56, 17.85%) and non-neoplastic proliferation hyperplasia/dysplasia (10/56, 17.85%). Complex carcinoma, carcinoma, and malignant myoepithelioma and malignant myoepithelioma were confirmed by p63 antibody. In these neoplasms, myoepithelial cells showed strong immunoreactivity with p63.

Polycystic liver in two adult llamas

Polycystic liver is usually considered an incidental finding in human and veterinary medicine. Two unrelated adult llamas ( Lama glama) with a history of marked anorexia and weight loss were received for autopsy and diagnostic workup. The main gross change in the liver of both animals was multiple variably sized cysts randomly distributed throughout the parenchyma. Histologically, the cysts compressed the adjacent parenchyma and were lined by a single layer of cuboidal-to-columnar epithelium, surrounded by a fibrous collagen capsule. The lumen of the cysts contained finely granular-to-homogeneous basophilic material. The lining epithelium displayed strong immunoreactivity for pancytokeratin AE1/AE3 and cytokeratins 7, 8, 8/18, and 19, and was negative for vimentin, confirming the biliary epithelial origin of the cysts. No parasitic or infectious agents, or neoplastic changes, were detected. All other laboratory tests performed in both llamas were negative or non-diagnostic, suggesting that the congenital hepatic cysts described may have been at least partly responsible for clinical disease in both animals.