Metabolomics Coupled with Pathway Analysis Provides Insights into Sarco-Osteoporosis Metabolic Alterations and Estrogen Therapeutic Effects in Mice

Postmenopausal osteoporosis (PMOP) and sarcopenia are common diseases that predominantly affect postmenopausal women. In the occurrence and development of these two diseases, they are potentially pathologically connected with each other at various molecular levels. However, the application of metabolomics in sarco-osteoporosis and the metabolic rewiring happening throughout the estrogen loss-replenish process have not been reported. To investigate the metabolic alteration of sarco-osteoporosis and the possible therapeutical effects of estradiol, 24 mice were randomly divided into sham surgery, ovariectomy (OVX), and estradiol-treated groups. Three-dimensional reconstructions and histopathology examination showed significant bone loss after ovariectomy. Estrogen can well protect against OVX-induced bone loss deterioration. UHPLC-Q-TOF/MS was preformed to profile semi- polar metabolites of skeletal muscle samples from all groups. Metabolomics analysis revealed metabolic rewiring occurred in OVX group, most of which can be reversed by estrogen supplementation. In total, 65 differential metabolites were identified, and pathway analysis revealed that sarco-osteoporosis was related to the alterations in purine metabolism, glycerophospholipid metabolism, arginine biosynthesis, tryptophan metabolism, histidine metabolism, oxidative phosphorylation, and thermogenesis, which provided possible explanations for the metabolic mechanism of sarco-osteoporosis. This study indicates that an UHPLC-Q-TOF/MS-based metabolomics approach can elucidate the metabolic reprogramming mechanisms of sarco-osteoporosis and provide biological evidence of the therapeutical effects of estrogen on sarco-osteoporosis.

Phosphate-buffered Formalin Fixative Under Controlled Fixation Temperature Yielded Excellently Preserved Histomorphology

Aim: Using mouse liver as experimental model, this study attempts to identify a formalin-based fixative and fixation temperature that jointly provides the best balance of preservation of tissue morphology. Methodology: Liver samples from fifty (50) albino mice aged between of 6 to 8 weeks consisting of both male and female was harvested following cervical dislocation and randomly distributed into control and experimental groups. Control samples were fixed in 10mL of 10% formalin at 25oC, 30oC, 35oC, 40oC, 45oC, 50oC, 55oC and 60oC respectively for 24 hours, while experimental samples were each fixed in equal volume of phosphate-buffered 10% formalin (pH 7.2, 7.4, 7.6 and 7.8) at the same temperature and time duration regimen and processed for general tissue morphology. Nuclear, cytoplasm and cell membrane morphology were assessed as evidence of the combined effectiveness of fixative and fixation temperature. Morphology was scored using a four-point grading scale with 1 being poor and 4 being excellent. Results: Nuclear, cytoplasm and cell membrane morphology were excellently preserved in tissue fixed with phosphate-buffered 10% formalin (pH 7.2) at 45oC. Tissue fixed with 10% formalin at 35oC exhibited excellent nuclear and cell membrane morphology, while excellent preservation of cell membrane morphology were observed in tissues fixed with 10% formalin at 40oC, phosphate-buffered 10% formalin (pH 7.4) at 55oC and 60oC, (pH 7.6) at 50oC and 55oC and (pH 7.8) at 55oC respectively. Furthermore, excellent preservation of nuclear morphology was observed in tissue fixed with phosphate-buffered 10% formalin (pH 7.8) at 60oC. Conclusion: Phosphate-buffered 10% formalin at a temperature of 45oC and pH 7.2 provide an excellent formalin-based fixative and fixation temperature that adequately preserves the microanatomy of tissue for histopathology examination.

Oral vaccination of piglets against Mycoplasma hyopneumoniae using silica SBA-15 as an adjuvant effectively reduced consolidation lung lesions at slaughter

Mycoplasma (M.) hyopneumoniae is the main pathogen of porcine enzootic pneumonia (PEP). Its controlling is challenging, and requires alternative strategies. This study aimed to develop an oral vaccine against M. hyopneumoniae using a nanostructured mesoporous silica (SBA-15) as an adjuvant, and compare its effect with an intramuscular (IM) commercial vaccine (CV). Fifty 24 day-old M. hyopneumoniae-free piglets composed five equal groups for different immunization protocols, consisting of a CV and/or oral immunization (OI). Control piglets did not receive any form of immunization. All piglets were challenged with M. hyopneumoniae strain 232 on D49 by tracheal route. IgA antibody response in the respiratory tract, bacterial shedding and serum IgG were evaluated. The piglets were euthanized on 28 (D77) and 56 (D105) days post-infection. Lung lesions were macroscopically evaluated; lung fragments and bronchoalveolar fluid (BALF) were collected for estimation of bacterial loads by qPCR and/or histopathology examination. All immunization protocols induced reduction on Mycoplasma-like macroscopic lung lesions. IgA Ab responses anti-M. hyopneumoniae, the expression of IL-4 cytokine and a lower expression of IL-8 were induced by CV and OI vaccines, while IgG was induced only by CV. Oral immunization using silica as a carrier-adjuvant can be viable in controlling M. hyopneumoniae infection.

Primary cavernous hemangioma of the glans penis with balanitis xerotica obliterans: a case report

The uniqueness of this case is in its rarity and divergent presentation. Here, we report a case of 61 years old diabetic male who presented with painless swelling on the glans penis for the past 6 months along with bleeding from the mass and difficulty to pass urine for 1 month. Examination revealed solid, mass arising from the tip of glans with non-retractile, indurated prepuce. No clinically palpable lymphadenopathy was present. Dorsal slit was performed and mass excised. Histopathology examination revealed cavernous hemangioma. Primary cavernous hemangioma of glans penis is very rare but challenging for urologists to diagnose and manage such cases.

A case report of male breast cancer in a 14-year-old patient

Breast cancer is one of the most common malignancies. This malignancy can originate from the epithelium of the ducts or lobules of the breast. Although rare, breast cancer can also be found in men. The incidence of breast cancer in men is <1% of the total breast cancer and it is estimated that 1 in 100.00 men worldwide is diagnosed with breast cancer. Breast cancer in pediatric patients is rare. Incidence of breast cancer in pediatric is estimated to be 0.2-0.8/100.000 for females with less than 20 years old. In this case report is a 14-years-old male with ten years history of a mass in his right breast. At first the mass was said to be as small as a marble, then since last year the mass have grown bigger. Biopsy and histopathology examination were performed to confirm the diagnosis. The result of histopathology examination was ductal invasive carcinoma. The patient and family refused to undergo chemotherapy. The incidence of breast cancer in pediatric is rare, and it has high mortality rate in pediatric patient.

Pathology and molecular characterization of Leucocytozoon caulleryi from backyard chickens in Khon Kaen Province, Thailand

Aim: The aim of this study was to characterize Leucocytozoon caulleryi from backyard chickens in Khon Kaen Province, Thailand. Materials and Methods: Tissue samples were collected from backyard chickens suspected to have leucocytozoonosis and subjected to histopathology examination. The BLAST Tool at NCBI GenBank (Basic Local Alignment Research Tools) (http://www.ncbi.nlm.nih.gov/BLAST) was used to identify the nucleotide sequence of the partial cytochrome c oxidase subunit I (cox I) gene. A Phylogenetic tree for analysis of L. caulleryi was constructed by using MEGA 7.0 software (https:// www.megasoftware.net/). Results: The necropsy results revealed the subcutaneous hemorrhages of pectoral muscles, multifocal hemorrhages of the thymus and pectoral muscles, hemorrhage of the proventriculus and peritoneal cavity, and megaloschizonts of the pancreas and intestine, including subcapsular hemorrhages of the liver. Microscopic examination revealed numerous megaloschizonts of Leucocytozoon spp. in the pectoral muscles, intestine, pancreas, and thymus. Molecular analysis of the partial cox I gene showed that the causal agent was closely related to L. caulleryi reported in Japan. Conclusion: From these results, L. caulleryi was determined to be the causal agent of leucocytozoonosis and was closely associated with L. caulleryi reported in Japan.

EP.WE.28Routine histopathology examination of the sleeved stomach after sleeve gastrectomy - significance and comparison with preoperative endoscopy

Background Sleeve gastrectomy (SG) is one of the most commonly performed bariatric procedures. However, the available literature on histological examination of the resected sleeved stomachs is widely inconsistent. This study aimed at assessing the histopathological findings after SG performed in our center compared to the routinely performed preoperative esophagogastroduodenoscopy (OGD) and the intraoperative findings. Material and methods We conducted a retrospective cohort study using the bariatric unit database of all patients who had SG procedures from June 2007 to December 2019. Results Out of the 620 SG performed, there were 219 patients with complete data, including preoperative OGD reports, operative notes, and postoperative histopathology reports. We found normal specimens in 43.83% (n = 96) and evidence of inflammation in 50.68% (n = 111) of cases. There were 10 (4.56%) specimens with evidence of inflammation and other lesions and two (0.91%) specimens with lesions only. Twenty-five patients had inflammation, and 10 had lesions on preoperative OGD with negative histopathology reports. Conclusion Routine histopathology examination after SG does not change the management, and preoperative OGD can detect inflammatory changes and lesions, which could have been missed with histology analysis only.

Mesenchymal chondrosarcoma in the maxillary gingiva of a Maltese dog: a case report

A 13-year-old castrated male Maltese dog was presented to a local animal hospital with an oral hemorrhage. An intraoral examination revealed an irregular proliferated lobular mass at the right side of the maxillary gingiva and hard palate. A surgically excised mass was requested for a histopathology examination. Histopathologically, the neoplastic foci were composed of biphasic morphologic patterns, such as primitive mesenchymal tissue and mature or immature cartilage tissue. Immunohistochemically, most of the neoplastic cells forming cartilaginous islands tested positive for S-100; the surrounding mesenchymal cells tested positive for vimentin. This paper describes a rare case of mesenchymal chondrosarcoma in the maxillary gingiva of a Maltese dog.

Cutaneous angiomatosis in a dog: a case report

A 1-year-old, castrated male mixed-breed dog presented with diffuse, purple lesions arranged in an irregular patchy pattern, with a slight elevation on the right hindlimb extending from the tarsus joint to the upper region of the thigh. Dermatological examinations and fungal and bacterial cultures revealed no infectious agents. The therapeutic response to antibiotics and antifungal agents was negative. A histopathology examination of the lesion revealed vascular proliferation with vasodilation and numerous varying-sized vessels. Mast-cell-dominated perivascular cuffing was also noted. The dog was diagnosed with cutaneous angiomatosis due to diffuse lesions and the histopathology findings of hemangioma.

Malignant Melanoma of Anal Canal

Anal melanoma (AM) is a rare and highly aggressive mucosal melanocytic malignancy. We report a case of 70 year old male presented with chief complaints of anal pain and intermittent rectal bleeding with clinical diagnosis of Polyp or Carcinoma. On histopathology examination malignant melanoma was suggested which was further confirmed by immunohistochemistry (S100 and HMB 45 strongly positive). Anal melanoma is rare neoplastic condition with catastrophic outcome.