Endoscopic ultrasound-guided fine-needle biopsy as a tool for studying the intra-tumoral microbiome in pancreatic ductal adenocarcinoma: a pilot study

A new approach by investigating the intra-tumoral microbiome raised great interest because they may influence the host immune response and natural history of the disease. However, previous studies on the intra-tumoral microbiome of pancreatic ductal adenocarcinoma (PDAC) were mostly based on examining the formalin-fixed paraffin-embedded tumor specimens. This study aims to investigate the feasibility of using endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) as a complementary procedure of surgical biopsy to obtain adequate fresh pancreatic cancer tissue for intra-tumoral microbial research. This was a prospective pilot study performed at a single tertiary referral center. We obtained pancreatic cancer tissue by EUS-FNB and surgical biopsy, respectively. We amplified the V3-V4 hyper-variable region of bacterial 16S ribosomal ribonucleic acid (rRNA) genes, constructed a pair-end library, and performed high-throughput sequencing. From August 2020 to November 2020, nine eligible patients with PDAC were enrolled in this study. The intra-tumoral microbiome profile was successfully generated from the PDAC cancer tissue obtained by EUS-FNB as well as by surgical biopsy. There was no significant difference in intra-tumoral alpha-diversity or bacterial taxonomic composition between tissues obtained by EUS-FNB and by surgical biopsy. EUS-FNB can collect sufficient fresh cancer tissue for microbiome analyses without complication. The intra-tumoral microbiome profile in tissues obtained by EUS-FNB had similar alpha-diversity and taxonomic profiles with those obtained by surgical biopsy. It implicated, except for surgical biopsy, EUS-FNB can be another valid and valuable tool for studying intra-tumoral microbiome in patients with resectable and unresectable PDAC.

Diagnostic Value of Vascular Endothelial Growth Factor C (VEGF-C) and CA 15-3 in Breast Cancer

ABSTRACT Background: expression of VEGF-C and CA 15-3 may be useful to differentiate between malignant and benign breast tumour because VEGF-C plays a role in promoting angiogenesis and lymphangiogenesis in malignant processes and CA 15-3 is the soluble form of transmembrane protein MUC1, a tumour marker which shows higher expression in breast cancer.Objective: to determine the diagnostic value of VEGF-C and CA 15-3 as tumour markers in patients with breast cancer.Methods: this diagnostic study recruited 76 patients that underwent surgical biopsy procedures at Dr. Kariadi and Pantiwilasa Citarum Hospitals Semarang. The VEGF-C and CA 15-3 levels in blood specimens taken before surgical biopsy procedure was determined using ELISA method. An ROC curve and AUC were used to establish the cut-off points and diagnostic value. Pathology examination results from the biopsy specimens were used as the gold standard.Results: the cut-off value for VEGF-C and CA 15-3 were 989.50 pg/mL and 74.00 U/mL. Sensitivity for VEGF-C, CA 15-3 and VEGF-C+CA 15-3 were 76.6%, 64.1% and 89.1%. Specificity for VEGF-C, CA 15-3 and VEGF-C+CA 15-3 were 75.0%, 75.0% and 50.0%. The AUC for VEGF-C, CA 15-3 and VEGF-C+CA 15-3 was 0.831 (95% CI = 0.727-0.934), 0.742 (95% CI = 0.628-0.856) and 0.840 (95% CI = 0.742-0.938).Conclusion: VEGF-C in combination with CA 15-3 is the best diagnostic parameter for breast cancer and has the best accuracy as a tumour marker for breast cancer.

ML-7 Liquid biopsy for MYD88 mutation in cerebrospinal fluid in patients with suspected primary CNS lymphoma

Background: Treatment intervention for central nervous system lymphoma (CNSL) requires pathological diagnosis by surgical biopsy. However, there are some cases in which the risk of surgery is high due to age, comorbidities, localization of lesions, etc. We are developing a CNSL diagnostic method based on the detection of MYD88 L265P mutation by digital PCR (dPCR) using CSF-DNA, and a high accuracy with a sensitivity of 92.9% and a specificity of 100% has been reported. Here, we report two cases with suspected brain stem CNSL, whose treatment strategy was determined by integrated clinico-laboratory information including neurological presentations, imaging, and the result of liquid biopsy. Result: Case 1. A 63-year-old woman visited our hospital with a complaint of right hemiplegia, which deteriorated in two months. MR images revealed a contrast-enhancing lesion in the left midbrain-ventral pons, suggesting CNSL. Biopsy was not considered because of its location, while dPCR using CSF-DNA showed a cluster of MYD88 mutation signals. Based on these work-ups, she was treated with high-dose methotrexate-based chemotherapy, resulting in a complete response with marked improvement of symptoms. Case 2. An 83-year-old man was referred for a history of diplopia and ataxic gait lasting for a month. MR images revealed an invasive lesion on his right midbrain-dorsal pons. Biopsy was declined due to the location, and liquid biopsy using CSF-DNA was performed to assist the diagnosis. In the first test, the CSF-DNA yield was too insufficient to determine the mutation signal by dPCR. The second dPCR using sufficient amount of CSF-DNA resulted in the Target/Total value of 0.049% which was lower than the threshold, suggesting the absence of MYD88 mutation. The patient underwent radiation therapy accordingly.Conclusions: CSF MYD88 mutation analysis by dPCR may have clinical utility and requires sufficient amount of CSF-DNA for exclusion of noise signals.

Prevalence and distribution of canine neoplastic and non-neoplastic cutaneous lesions in Serbia: a retrospective study of 2432 Cases (2011 – Mid 2021)

Cutaneous lesions, especially skin tumors in dogs, are among the most common lesions in this animal species. The aim of this study was to identify the most common types of canine cutaneous lesions, to determine the absolute and relative frequency of each type of cutaneous lesion, anatomical locations, mean age, as well as gender and breed distribution. The examination included all samples of cutaneous lesions in dogs obtained by surgical biopsy in veterinary clinics and examined at the Laboratory of the Department of Pathology at the Faculty of Veterinary Medicine, University of Belgrade from the 1st January 2011 to the 1st July 2021. In this period (126 months), a total of 2432 samples of cutaneous lesions were examined, of which 1984 (81.58%) were tumors (1037/1984, 52.27% benign and 947/1984, 47.73% malignant) and 448 (18.42%) non-neoplastic cutaneous lesions. The most commonly found cutaneous tumors were: mast cell tumor (17.34% of all tumors), histiocytoma (9.78%), papilloma (7.91%), lipoma (7.81%), squamous cell carcinoma (7.36%), trichoblastoma (4.44%), hepatoid adenoma (4.39%) and malignant melanoma (4.18%). The most common non-neoplastic cutaneous lesions were: follicular cyst(s) (35.04% of all non-neoplastic lesions), pyogranulomatous chronic dermatitis (23.88%), lymphocytic dermatitis (7.37%), hyperkeratosis (4.24%), and granulomatous dermatitis (3.79%). Our results substantially confirm previously reported data regarding cutaneous neoplastic and nonneoplastic lesions in dogs, and provide updated information on their frequency, animal age, anatomic location and breed distributions.

Determining chromosomal arms 1p/19q co-deletion status in low graded glioma by cross correlation-periodogram pattern analysis

Prediction of mutational status of different graded glioma is extremely crucial for its diagnosis and treatment planning. Currently FISH and the surgical biopsy techniques are the ‘gold standard’ in the field of diagnostics; the analyses of which helps to decide appropriate treatment regime. In this study we proposed a novel approach to analyze structural MRI image signature pattern for predicting 1p/19q co-deletion status non-invasively. A total of 159 patients with grade-II and grade-III glioma were included in the analysis. These patients earlier underwent biopsy; the report of which confirmed 57 cases with no 1p/19q co-deletion and 102 cases with 1p/19q co-deletion. Tumor tissue heterogeneity was investigated by variance of cross correlation (VoCC). Significant differences in the pattern of VoCC between two classes was quantified using Lomb-Scargle (LS) periodogram. Energy and the cut-off frequency of LS power spectral density were derived and utilized as the features for classification. RUSBoost classifier was used that yield highest classification accuracy of 84% for G-II and 87% for G-III glioma respectively in classifying 1p/19q co-deleted and 1p/19q non-deleted glioma. In clinical practice the proposed technique can be utilized as a non-invasive pre-confirmatory test of glioma mutation, before wet-lab validation.

IgG4-related chronic sclerosing sialadenitis in a child with recurrent parotitis: a case report

Background IgG4-related disease (IgG4-RD) includes a group of immune-mediated diseases histologically characterized by lymphoplasmacytic infiltrate with a prevalence of IgG4-positive plasma cells, storiform fibrosis and obliterative phlebitis. Autoimmune pancreatitis, sialadenitis, dacryoadenitis and retroperitoneal fibrosis are the most frequent manifestations. IgG4-related sialadenitis usually affects submandibular glands and is very rare in children. Here we report the case of IgG4-related sialadenitis in a six-year-old patient previously diagnosed as juvenile recurrent parotitis. Case presentation A six-year-old patient was referred to our Centre for left parotid swelling of 4 × 3 cm, that was tender, soft in consistency, with overlying red and warm skin. His general condition was good but he was subfebrile; general examination revealed mild enlargement of left cervical lymph nodes. In the last 2 years he had had five episodes of parotitis, diagnosed by another pediatric Center as juvenile recurrent parotitis. On ultrasound examination the left parotid gland appeared enlarged, inhomogeneous, with a colliquative intraparotid lymph node and no evidence of sialolithiasis. Laboratory tests showed an increase of white blood cells and anti-VCA IgM and IgG positivity, with anti-EBNA e anti-EA I negativity. The patient was initially treated with oral antibiotics, but after 10 days the parotid became fluctuating, requiring surgical biopsy and drainage. Postoperative course was regular, with complete remission under oral antibiotic and steroid therapy. Microbiological tests, including cultures for aerobic and anaerobic bacteria, mycobacteria and Bartonella, were negative. Surprisingly, histology showed marked fibrosis and histiocytic and lymphoplasmacellular infiltrate with polyclonal plasma cells mostly expressing IgG4 immunoglobulins. Thus, the diagnosis of IgG4 related chronic sialadenitis in recurrent parotitis and recent EBV infection was made. Conclusions IgG4-related sialadenitis is very unusual in children. Histology plays a key role in diagnosis, considering that up to 30% of patients have normal serum IgG4 levels, as shown in our case. The lack of previous histological data makes it impossible to attribute our patient’s previous episodes of parotitis to IgG4-RD, though it is a very consistent possibility.

Post-Mortem Brain Donations vs Pre-Mortem Surgical Resections for Glioblastoma Research: Viewing the Matter as a Whole

There have been limited improvements in diagnosis, treatment and outcomes of primary brain cancers, including glioblastoma, over the past 10 years. This is largely attributable to persistent deficits in understanding brain tumour biology and pathogenesis due to a lack of high-quality biological research specimens. Traditional, pre-mortem, surgical biopsy samples do not allow full characterisation of the spatial and temporal heterogeneity of glioblastoma, nor capture end-stage disease to allow full evaluation of the evolutionary and mutational processes that lead to treatment resistance and recurrence. Furthermore, the necessity of ensuring sufficient viable tissue is available for histopathological diagnosis, while minimising surgically induced functional deficit, leaves minimal tissue for research purposes and results in formalin fixation of most surgical specimens. Post-mortem brain donation programs are rapidly gaining support due to their unique ability to address the limitations associated with surgical tissue sampling. Collecting, processing, and preserving tissue samples intended solely for research provides both a spatial and temporal view of tumour heterogeneity as well as the opportunity to fully characterise end-stage disease from histological and molecular standpoints. This review explores the limitations of traditional sample collection and the opportunities afforded by post-mortem brain donations for future neurobiological cancer research.

Intraoperative Ultrasound Applications in Intracranial Surgery

The methods and use of intraoperative ultrasound in 33 canine and five feline patients and its ability to localize and identify anatomical structures and pathological lesions in canines and felines undergoing intracranial surgery are described from a case series. All were client-owned referral patients admitted for neurologic evaluation, with an advanced imaging diagnosis of an intracranial lesion, and underwent surgical biopsy or surgical removal of the lesion. Medical records, retrieval and review of imaging reports, and characterization of findings for all canine and feline patients show that intraoperative ultrasound guidance was used in intracranial procedures during the period of 2012 and 2019. Twenty-nine of the canine patients had intracranial tumors. The remainder had various other conditions requiring intracranial intervention. Three of the feline patients had meningiomas, one had a depressed skull fracture, and one had an epidural hematoma. The tumors appeared hyperechoic on intraoperative ultrasound with the exception of cystic portions of the masses and correlated with the size and location seen on advanced imaging. Statistical comparison of the size of images seen on ultrasound and on MRI for 20 of the canine tumors revealed no statistical differences. Neuroanatomical structures, including vascular components, were easily identified, and tumor images correlated well with preoperative advanced imaging. The authors conclude that intraoperative ultrasound is a valuable asset in intracranial mass removals and can augment surgical guidance in a variety of intracranial disorders that require surgery. This is the first known publication in veterinary surgery of using intraoperative ultrasound as a tool in the operating theater to identify, localize, and monitor the removal/biopsy of intracranial lesions in small animals undergoing craniotomy/craniectomy.

Desmoplastic fibroma of bone: A morphological and immunohistochemical characterization

Background: Desmoplastic Fibroma (DF) of bone is a locally aggressive and infrequent benign neoplasm. Recently was described a role of vascular endothelial growth factor in the interstitial fibrotic processes. Case presentation: A 13-year-old female presented with pain, swelling and limitation of movements in right forearm. An osteolytic lesion at the distal end of the right radius was shown, with pathologic concentration of Technetium 99 and slight enhancement of soft tissue lesion employing computerized axial tomography. The surgical biopsy showed nodular formations of hyalinized collagen fibers arranged in thick bands with few well-differentiated interstitial fibroblasts / myofibroblasts, focally expressing VEGF-A. Conclusion: The intramedullary neoplastic proliferation is limited by the cortical bone, provoking compression of the intratumorally micro-vessels, favoring both, the extracellular matrix and VEGF-A synthesis. Future research should include therapeutic intervention with anti-CD117 and anti-VEGF-A drugs, with the aim of limiting tumor growth, facilitating the complete surgical excision of the neoplasm. Keywords: desmoplastic fibroma; vascular endothelial growth factor; hyalinization; neoplasm progression.