The mutational frequency of BRAF and KRAS in low-grade serous testicular neoplasms-a case series.

AIMS: Low-grade serous neoplasms of the testis are rare neoplasms that show striking morphological similarities with the better-understood ovarian neoplasms. This study is to see if there are similar molecular abnormalities in these two tumours. The cell of origin, relationship with serous ovarian tumour and the pathogenesis of these neoplasms are not fully established. METHODS AND RESULTS: As low-grade serous ovarian neoplasms are known to harbour mutations in the MAPK pathway, we investigated the involvement of BRAF and KRAS mutations in low-grade testicular serous tumour by performing mutational analysis of seven cases. Mutational analysis was performed by melting curve analysis followed by bidirectional sequencing. Our findings showed BRAF and/or KRAS mutations in three of the seven cases, which is similar to the proportions reported in low-grade ovarian serous neoplasms. Of these three cases, one showed co-mutation of BRAF and KRAS. CONCLUSION: The findings of this study are in support of a role of aberrant signalling of the MAPK pathway in the pathogenesis of low-grade serous testicular neoplasms, and provide a genetic link between low-grade testicular and ovarian serous tumours.

Algal macromolecular mediated synthesis of nanoparticles for their application against citrus canker for food security.

Citrus canker is a disease of economic importance and there are limited biocontrol agents available to mitigate it in an integrated manner. This study was conducted to combat citrus canker disease using biologically active nanoparticles (Ag, Cu and ZnO and 300, 900, 1200, and 1500 ppm) synthesized from macromolecules extracted from alga, Oedogonium sp. The synthesis of the nanoparticles was confirmed by UV-Vis Spectroscopy, FTIR, SEM, XRD, and DLS Zeta sizer while their efficacy was tested against Xanthomonas citri by measuring zone of inhibition. Results indicated that Ag and Cu nanoparticles at 1200 ppm exhibit the highest activity against Xanthomonas citri, followed by ZnO at 1500 ppm. The minimum inhibitory concentrations (MIC) of Ag, Cu and ZnO NPs were 1, 2 and 10 mg mL-1, respectively while minimum bactericidal concentrations (MBC) were for Ag and Cu 2, 4 mg mL-1 and for ZnO NPs more then 10 mg mL-1, were required to kill the X. citri. Bacterial growth respectively. Macromolecules extracted from algal sources can produce nanoparticles with bactericidal potential, in the order of Ag > Cu > ZnO to mitigate citrus canker disease and ensuring sustainable food production amid the growing human population.

P2X7 receptor-mediated purinergic signaling promotes liver injury in acetaminophen hepatotoxicity in mice.

Inflammation contributes to liver injury in acetaminophen (APAP) hepatotoxicity in mice and is triggered by stimulation of immune cells. The purinergic receptor P2X7 is upstream of the nod-like receptor family, pryin domain containing-3 (NLRP3) inflammasome in immune cells and is activated by ATP and NAD that serve as damage-associated molecular patterns. APAP hepatotoxicity was assessed in mice genetically deficient in P2X7, the key inflammatory receptor for nucleotides (P2X7-/-), and in wild-type mice. P2X7-/- mice had significantly decreased APAP-induced liver necrosis. In addition, APAP-poisoned mice were treated with the specific P2X7 antagonist A438079 or etheno-NAD, a competitive antagonist of NAD. Pre- or posttreatment with A438079 significantly decreased APAP-induced necrosis and hemorrhage in APAP liver injury in wild-type but not P2X7-/- mice. Pretreatment with etheno-NAD also significantly decreased APAP-induced necrosis and hemorrhage in APAP liver injury. In addition, APAP toxicity in mice lacking the plasma membrane ecto-NTPDase CD39 (CD39-/-) that metabolizes ATP was examined in parallel with the use of soluble apyrase to deplete extracellular ATP in wild-type mice. CD39-/- mice had increased APAP-induced hemorrhage and mortality, whereas apyrase also decreased APAP-induced mortality. Kupffer cells were treated with extracellular ATP to assess P2X7-dependent inflammasome activation. P2X7 was required for ATP-stimulated IL-1ß release. In conclusion, P2X7 and exposure to the ligands ATP and NAD are required for manifestations of APAP-induced hepatotoxicity.

A Feasibility Study of Multisite Networked Digital Pathology Reporting in England.

BACKGROUND: The objective of the project was to evaluate the feasibility of introducing a single-networked digital histopathology reporting platform in the Southwest Peninsula region of England by allowing pathologists to experience the technology and recording their perceptions. This information was then used in planning future service development. The project was funded by the National Health Service (NHS) Peninsula Cancer Alliance and took place in 2020 during the COVID-19 pandemic. MATERIALS AND METHODS: Digital slides of 500 cases from Taunton were reported remotely in Truro, Plymouth, Exeter, Bristol, or Bath by using a single remote reporting platform located on the secure Health and Social Care Network (HSCN) that links NHS sites. These were mainly small gastrointestinal, skin, and gynecological specimens. The digital diagnoses were compared with the diagnoses issued on reporting the glass slides. At the end of the project, the pathologists completed a Google Forms questionnaire of their perceptions of digital pathology. The results were presented at a meeting with the funder and discussed. RESULTS: From the 500 cases there were nine cases of significant diagnostic discrepancy, seven of which involved the misrecognition of Helicobacter pylori in gastric biopsies. The questionnaire at the end of the project showed that there was a general agreement that the platform was easy to use, and the image quality was acceptable. It was agreed that extra work, such as deeper levels, was easy to request on the software platform. Most pathologists did not agree that digital reporting was quicker than glass slide reporting. Some were less confident in their digital diagnoses than glass diagnoses. They agreed that some types of specimens cannot easily be reported digitally. All users indicated that they would like to report at least half of their work digitally in the future if they could, and all strongly agreed that digital pathology would improve access to expert opinions, teaching, and multidisciplinary meetings. It was difficult to find pathologists with time to undertake remote digital reporting, in addition to their existing commitments. CONCLUSIONS: Overall, the pathologists developed a positive perception of digital pathology and wished to continue using it.

Interaction between GRP78 and IGFBP-3 Affects Tumourigenesis and Prognosis in Breast Cancer Patients.

Insulin-like growth factor binding protein 3 (IGFBP-3) plays a key role in breast cancer progression and was recently shown to bind to the chaperone protein glucose-regulated protein 78 (GRP78); however, the clinical significance of this association remains poorly investigated. Here we report a direct correlation between the expression of GRP78 and IGFBP-3 in breast cancer cell lines and tumour sections. Kaplan-Meier survival plots revealed that patients with low GRP78 expression that are positive for IGFBP-3 had poorer survival rates than those with low IGFBP-3 levels, and we observed a similar trend in the publicly available METABRIC gene expression database. With breast cancer cells, in vitro IGFBP-3 enhanced induced apoptosis, however when GRP78 expression was silenced the actions of IGFBP-3 were switched from increasing to inhibiting ceramide (C2)-induced cell death and promoted cell invasion. Using immunofluorescence and cell surface biotinylation, we showed that knock-down of GRP78 negated the entry of IGFBP-3 into the cells. Together, our clinical and experimental results suggest that loss of GRP78 reduces IGFBP-3 entry into cells switching its actions to promote tumorigenesis and predicts a poor prognosis in breast cancer patients.

What is the significance of pT0 at cystectomy?

The current treatment for muscle invasive bladder cancer is radical cystectomy. However, this approach leads to significant changes in the patient's quality of life, as well as potential treatment failure. And so, with the introduction of alternative and neoadjuvant therapies available, the presence of pT0 tumour at cystectomy confers a number of issues. The variability in numbers of pT0 tumours at cystectomy highlights the importance of adequate clinical staging, as well as the increasing successful use of neoadjuvant chemotherapy. Although current literature is limited, patients with prior clinical stage of muscle-invasive but node-negative disease are likely to demonstrate the most improvement in survivability if they subsequently develop pT0 at cystectomy. This review also highlights the importance of response to chemotherapy as an indicator of subsequent prognosis. With increasing numbers of pT0 tumours seen at radical cystectomy, it is suggested that more conservative measures, such as re-staging following neoadjuvant chemotherapy and even 'selective bladder preservation' treatment, may be the future for the management of muscle invasive bladder cancer.

Isolated metastasis of testicular seminoma to extralobar pulmonary sequestration.

This case report describes a 46-year-old patient with a very unusual pattern of isolated metastasis to an extralobar pulmonary sequestration following orchidectomy for seminoma. The patient underwent uncomplicated video-assisted thoracoscopic resection of the metastasis, involving the whole of the sequestration mass.

Expression of lymphotoxin-beta (LT-beta) in chronic inflammatory conditions.

Functional studies in gene-knockout and transgenic mice systems have shown that lymphotoxin-alpha and lymphotoxin-beta (LT-alpha and LT-beta) are of fundamental importance in peripheral lymphoid organ development, but it remains unclear what role these cytokines have to play in the adult immune response and in the pathogenesis of disease. In this study, a polyclonal anti-serum to human LT-beta was used to investigate the distribution of LT-beta by immunohistochemistry in normal and diseased tissues. In the gut, lymph nodes, spleen, and tonsil, there was some LT-beta present on a variety of lymphoid cell types. In contrast, strong staining for LT-beta was observed on plasma cells and a subpopulation of CD4+ T cells in tissues affected by chronic inflammatory disease or infection, for example in inflammatory bowel disease, and in lymph nodes obtained from patients with sarcoidosis and tuberculosis. In tuberculous and sarcoid lymph nodes, LT-beta expression also occurred on some but not all epithelioid histiocytes within granulomas and on multi-nucleated giant cells. These findings support a role for LT-beta in human disease and suggest that it might represent a therapeutic target in a variety of common infective or inflammatory disorders.