Endothelial cells do not arise from tumor-initiating cells in human hepatocellular carcinoma

BACKGROUND: Conventional models of carcinogenesis suggest that tumors recruit blood vessel formation from normal host tissues. This concept has recently been challenged by prominent studies of glioblastoma, which suggest that intratumoral endothelial cells (ECs) may arise from cancer stem cells/tumor-initiating cells (TICs). Hepatocellular carcinoma (HCC) is a common, highly vascularized tumor with few effective therapies, against which anti-angiogenic strategies are being actively explored. TICs are felt to play a role in HCC pathobiology, but their contributions to tumor vasculature have not been studied. METHODS: We examined human HCCs in settings that selected for tumor formation from functionally defined TICs, and in which the origin of intratumoral ECs from TICs as opposed to host tissues could be clearly distinguished. We generated HCC nodules in the livers of immunodeficient mice by intrasplenic injection of HCC cells from cell lines and patient specimens and studied the tumor ECs by immunohistochemistry for mouse and human markers. We then used immunohistochemistry for EC markers in combination with fluorescence in situ hybridization (FISH) for X and Y chromosomes to study the endothelium of recurrent HCC specimens resected from sex-mismatched liver allografts of patients who had undergone liver transplantation for HCC. RESULTS: We observed that all ECs in intrahepatic human HCC xenografts expressed mouse rather than human CD31. FISH analysis of recurrent HCCs resected from patients with sex-mismatched liver allografts revealed that all CD31(+) and CD34(+) intratumoral ECs originated from the donor allograft rather than the tumor. CONCLUSIONS: These observations suggest that the vasculature of human HCC arises from normal host tissues rather than from TICs, supporting ongoing efforts to target angiogenesis in HCC as it is currently understood, and suggesting that the contribution of TICs to the vasculature of other cancers is disease-specific

Modeling the effect of entrained sand particles on pressure transverse in a flowing gas well

Purpose The production of natural gas from the reservoir is always associated with entrained solid particle of different sizes mainly sand particles and crystalline salts. Entrained solid transport along the gas phase has been a great concern for gas production engineer, as the detrimental consequences are often associated to a desirable high operational parameters such rate and pressure transverse in producing well. Design/methodology/approach A variety of models for predicting pressure transverse in flowing gas wells have been reported in the literatures. Most of the models were based on steady state fluid flow equation that did not consider time factor which results in inaccurate at early production time. Some of the early investigators overlooked the effect of the entrained solid on the pressure transverse phenomena in a gas well. Hence, there is a need for developing a more realistic model for estimating pressure transverse at all times in flowing solid-gas vertical well. Findings This study presents equation for pressure drop in flowing vertical well without neglecting any term in the momentum equation by the inclusion of accumulation and kinetic term. The solution of the resulting differential equation gives functional relationship between solid-gas flow rates and pressure at any point in flowing well at any given production time

Human Solid Tumor Xenografts in Immunodeficient Mice Are Vulnerable to Lymphomagenesis Associated with Epstein-Barr Virus

Xenografting primary human solid tumor tissue into immunodeficient mice is a widely used tool in studies of human cancer biology; however, care must be taken to prove that the tumors obtained recapitulate parent tissue. We xenografted primary human hepatocellular carcinoma (HCC) tumor fragments or bulk tumor cell suspensions into immunodeficient mice. We unexpectedly observed that 11 of 21 xenografts generated from 16 independent patient samples resembled lymphoid neoplasms rather than HCC. Immunohistochemistry and flow cytometry analyses revealed that the lymphoid neoplasms were comprised of cells expressing human CD45 and CD19/20, consistent with human B lymphocytes. In situ hybridization was strongly positive for Epstein-Barr virus (EBV) encoded RNA. Genomic analysis revealed unique monoclonal or oligoclonal immunoglobulin heavy chain gene rearrangements in each B-cell neoplasm. These data demonstrate that the lymphoid neoplasms were EBV-associated human B-cell lymphomas. Analogous to EBV-associated lymphoproliferative disorders in immunocompromised humans, the human lymphomas in these HCC xenografts likely developed from reactivation of latent EBV in intratumoral passenger B lymphocytes following their xenotransplantation into immunodeficient recipient mice. Given the high prevalence of latent EBV infection in humans and the universal presence of B lymphocytes in solid tumors, this potentially confounding process represents an important pitfall of human solid tumor xenografting. This phenomenon can be recognized and avoided by routine phenotyping of primary tumors and xenografts with human leukocyte markers, and provides a compelling biological rationale for exclusion of these cells from human solid tumor xenotransplantation assays

Banff 2022 liver group meeting report: monitoring long term allograft health.

The Banff Working Group on Liver Allograft Pathology met in September 2022. Participantsincluded hepatologists, surgeons, pathologists, immunologists and histocompatibility specialists.Presentations and discussions focused on the evaluation of long-term allograft health, including noninvasive and tissue monitoring, immunosuppression optimisation and long-term structural changes.Potential revision of the rejection classification scheme to better accommodate and communicate lateT cell-mediated rejection patterns and related structural changes, such as nodular regenerativehyperplasia, were discussed. Improved stratification of long-term maintenance immunosuppression tomatch the heterogeneity of patient settings will be central to improving long-term patient survival.Such personalised therapeutics are in turn contingent on better understanding and monitoring ofallograft status within a rational decision-making approach, likely to be facilitated in implementationwith emerging decision support tools. Proposed revisions to rejection classification emerging fromthe meeting include incorporation of interface hepatitis and fibrosis staging. These will be opened toonline testing, modified accordingly and subject to consensus discussion leading up to the next Banffconference

A Fatal Case of Diffuse Alveolar Hemorrhage in the Setting of Systemic Lupus Erythematosus: A Case Report and Review of Noninfectious Causes of Acute Pulmonary Hemorrhage in Adults

Systemic lupus erythematosus (SLE) is an autoimmune connective tissue disease, characterized by autoantibody production and immune complex formation, that has the potential to affect virtually any organ. Pleuropulmonary involvement occurs in 50–70% and commonly manifests as pleuritis and pleural effusion. Diffuse alveolar hemorrhage (DAH) is a rare manifestation of SLE. Most cases of DAH occur in young adults with an underlying autoimmune disease such as systemic vasculitis or Goodpasture syndrome. SLE is typically lower on the list of initial differential diagnoses of DAH due to its rarity compared to other etiologies. We present a case of a patient with dyspnea on exertion, dry coughs, lower extremity edema, and intermittent periorbital edema who ultimately succumbed to respiratory failure secondary to DAH in the setting of SLE. The diagnosis of SLE was suspected clinically and confirmed at autopsy due to her rapid clinical deterioration. DAH requires prompt intervention, and management is guided by the underlying disease process. SLE is a potentially treatable disease; therefore, timely diagnosis is important in order to exclude other noninfectious causes of DAH (reviewed in this report) and to initiate appropriate therapy

Patient demographics, parent HCC grade, and xenograft characteristics.

a<p>Degree of tumor differentiation documented in clinical pathology report.</p>b<p>Number of days between implantation of tumor sample and harvesting of a 1.5 cm3 xenograft.</p>c<p>Both xenografts demonstrated similar histology.</p>d<p>Non-alcoholic steatohepatitis.</p

Expression of leukocyte markers and EBER in non-HCC-like xenografts.

<p>(A) Representative section (×200) from a parent HCC sample that gave rise to a non-HCC-like xenograft, demonstrating a typical distribution of CD45⁺ leukocytes along a portal tract invaded by the tumor, only a small fraction of which are CD20⁺ B lymphocytes; EBER ISH is negative. (B) Representative sections (×400) from three non-HCC-like xenografts demonstrating that a very high proportion of cells stain positively for human CD45 and human CD20 (brown), consistent with human B lymphocytes; EBER ISH is very strongly positive in the cells in these xenografts (dark blue). (C) Representative multiparameter flow cytometry analysis of freshly isolated cells from a non-HCC-like xenograft demonstrating that a large proportion of tumor cells are human CD45⁺ leukocytes (left plot), and that the majority of the gated CD45⁺ population also expresses human CD19⁺ (right plot), consistent with B lymphocytes.</p

Immunoglobulin heavy chain gene rearrangements in non-HCC-like xenografts.

<p>PCR amplification of the variable region of the human IgH gene demonstrating unique dominant rearrangements in all of the non-HCC-like xenografts, confirming clonal B-cell proliferation. Dominant rearrangements were not amplified in HCC-like xenografts. Successful amplification of the β-globin gene confirms integrity of the genomic DNA analyzed.</p