190 THE USEFULNESS OF A BILATERAL TRANS-CERVICAL PNEUMOMEDIASTINAL APPROACH FOR MEDIASTINOSCOPIC RADICAL ESOPHAGECTOMY

Several authors have reported on the left trans-cervical and transhiatal approaches under pneumomediastinum and right cervical open surgery for mediastinoscopic esophagectomy. However, with these approaches, sufficient dissection of the right upper mediastinal paraesophageal lymph nodes, right recurrent nerve lymph nodes and the subaortic arch to the left tracheobronchial lymph nodes is thought to be difficult. We herein report the usefulness of the ‘bilateral’ trans-cervical pneumomediastinal approach. Methods Ten patients with thoracic esophageal cancer were treated using this approach. Under pneumomediastinum via a right neck incision, the right cervical and upper mediastinal paraesophageal lymph nodes were dissected. The left recurrent nerve lymph nodes were dissected using a left trans-cervical pneumomediastinal approach. The subaortic arch to the left tracheobronchial lymph nodes was dissected with a combined right and left trans-cervical crossover approach. After this approach, thoracoscopic observation was then performed in the left decubitus position, and if the lymph nodes were not sufficiently dissected, the remnant lymph nodes were retrieved thoracoscopically. Results The average total number of dissected lymph nodes among the right cervical and upper mediastinal paraesophageal lymph nodes identified with a right cervical open/right trans-cervical mediastinoscopic/right thoracoscopic approach was 3.2/4.0/0.6, respectively. The average total number of dissected lymph nodes among the subaortic arch to the left tracheobronchial lymph nodes with a right trans-cervical mediastinoscopic/right thoracoscopic approach was 1.5/0.6, respectively. These findings indicate that, without the right trans-cervical pneumomediastinal approach, roughly four of the right cervical and upper mediastinal paraesophageal lymph nodes and one or two of the subaortic arch to the left tracheobronchial lymph nodes could not have been retrieved. Conclusion A bilateral trans-cervical pneumomediastinal approach is useful for achieving sufficient upper mediastinal lymph node dissection and esophagectomy.

The Post-Acute Phase of SARS-CoV-2 Infection in Two Macaque Species Is Associated with Signs of Ongoing Virus Replication and Pathology in Pulmonary and Extrapulmonary Tissues

The post-acute phase of SARS-CoV-2 infection was investigated in rhesus (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis). During the acute phase of infection, SARS-CoV-2 was shed via the nose and throat, and viral RNA was occasionally detected in feces. This phase coincided with a transient change in systemic immune activation. Even after the alleged resolution of the infection, computed tomography (CT) and positron emission tomography (PET)-CT revealed pulmonary lesions and activated tracheobronchial lymph nodes in all animals. Post-mortem histological examination of the lung tissue revealed mostly marginal or resolving minimal lesions that were indicative of SARS-CoV-2 infection. Evidence for SARS-CoV-2-induced histopathology was also found in extrapulmonary tissue samples, such as conjunctiva, cervical, and mesenteric lymph nodes. However, 5–6 weeks after SARS-CoV-2 exposure, upon necropsy, viral RNA was still detectable in a wide range of tissue samples in 50% of the macaques and included amongst others the heart, the respiratory tract and surrounding lymph nodes, salivary gland, and conjunctiva. Subgenomic messenger RNA was detected in the lungs and tracheobronchial lymph nodes, indicative of ongoing virus replication during the post-acute phase. These results could be relevant for understanding the long-term consequences of COVID-19 in humans.

MALDI MSI Reveals the Spatial Distribution of Protein Markers in Tracheobronchial Lymph Nodes and Lung of Pigs after Respiratory Infection

Respiratory infections are a real threat for humans, and therefore the pig model is of interest for studies. As one of a case for studies, Actinobacillus pleuropneumoniae (APP) caused infections and still worries many pig breeders around the world. To better understand the influence of pathogenic effect of APP on a respiratory system—lungs and tracheobronchial lymph nodes (TBLN), we aimed to employ matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI). In this study, six pigs were intranasally infected by APP and two were used as non-infected control, and 48 cryosections have been obtained. MALDI-TOF MSI and immunohistochemistry (IHC) were used to study spatial distribution of infectious markers, especially interleukins, in cryosections of porcine tissues of lungs (necrotic area, marginal zone) and tracheobronchial lymph nodes (TBLN) from pigs infected by APP. CD163, interleukin 1β (IL-1β) and a protegrin-4 precursor were successfully detected based on their tryptic fragments. CD163 and IL-1β were confirmed also by IHC. The protegrin-4 precursor was identified by MALDI-TOF/TOF directly on the tissue cryosections. CD163, IL-1β and protegrin-4 precursor were all significantly (p < 0.001) more expressed in necrotic areas of lungs infected by APP than in marginal zone, TBLN and in control lungs.

Tuberculosis in wild South American sea lions Otaria flavescens stranded in Chubut, Argentina

Pinniped tuberculosis, commonly caused by Mycobacterium pinnipedii, is a zoonotic disease reported in free-living and captive otariid species of the southern hemisphere. Currently, data concerning pinniped tuberculosis in South America are scarce, reinforcing the need for further studies of the disease in free-ranging pinnipeds. In this study, we investigated the presence of tuberculosis in South American sea lions Otaria flavescens (SASLs) stranded along the Chubut coastline (Argentina). Necropsies were performed in 9 SASLs, and tissue samples were collected for histopathology, bacteriology, and molecular diagnosis. Four SASLs showed enlarged tracheobronchial lymph nodes (TBLNs) with multifocal to coalescing granulomas. In these animals, a direct IS6110-PCR amplification confirmed the presence of a Mycobacterium tuberculosis complex member in TBLNs (n = 4) and lungs (n = 2), but the agent could not be further identified. In one SASL, Mycobacterium murale was isolated from lungs without lesions. This study confirms the presence of tuberculosis in SASLs from Chubut, where tourist activities promote close interaction with the animals, generating a potential risk to human health. Further research is currently focusing on addressing the prevalence of tuberculosis in wild SASLs, to assess the risk for public health and develop management strategies to avoid human infection.

Metastatic renal interstitial cell tumor in a dog

Renal interstitial cell tumor (RICT) is a rare renal sarcoma of dogs that arises from renal interstitial cells. Herein we describe a RICT in an 8-y-old female Labrador Retriever dog that died after a 2-d history of lethargy and disorientation. Grossly, soft white nodules of 1–10 mm diameter were present in the renal cortex and corticomedullary junction of both kidneys, left cardiac ventricular wall, and right cerebral hemisphere. A pale-white to yellow, firm, irregular mass effaced 80% of the right pulmonary parenchyma, involving mainly the cranial and middle lobes, and the adjacent tracheobronchial lymph nodes. Histologically, the renal, myocardial, and cerebral neoplasm consisted of interlacing bundles of stellate-to-spindle cells with eosinophilic vacuolated cytoplasm and round-to-oval nuclei with finely stippled chromatin. The mitotic count was 28 per 2.37 mm2. Alcian blue stain revealed an extracellular myxomatous matrix throughout the neoplasm. Neoplastic cells had cytoplasmic immunolabeling for vimentin and cyclooxygenase 2. The pulmonary and tracheobronchial neoplasm consisted of infiltrative nodules of cuboidal epithelial cells that had a moderate amount of eosinophilic cytoplasm and round nuclei with coarsely stippled chromatin. There were 5 mitoses per 2.37 mm2. Neoplastic cells had cytoplasmic and nuclear immunolabeling for cytokeratin AE1/AE3 and thyroid transcription factor 1, respectively. Morphologic and immunohistochemical findings were consistent with a RICT with cardiac and cerebral metastases, and a pulmonary carcinoma with tracheobronchial lymph node metastasis.

Blastomyces dermatitidis pneumonia in a llama

A 20-y-old female llama ( Lama glama) was euthanized after a history of chronic dyspnea and osteoarthritis. At autopsy, the lungs were covered by clear gelatinous material and expanded by firm, variably discrete, tan-white nodules up to 8 cm diameter containing tan-white, viscous material. The tracheobronchial lymph nodes were firm and enlarged up to 6 × 4 × 3 cm; the thoracic aorta and carotid arteries were lined by hard, tan-white, mineralized intimal plaques. Histologic examination of lung revealed numerous 10–20 μm diameter yeasts with clear 1–2 μm thick double-contoured walls, central basophilic nuclei, and frequent broad-based budding, consistent with Blastomyces dermatitidis. DNA sequencing confirmed the diagnosis. B. dermatitidis should be considered in the differential diagnosis of pulmonary disease in llamas.