Objective To evaluate viral loads at different stages of disease progression in patients infected with the 2019 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the first four months of the epidemic in Zhejiang province, China. Design Retrospective cohort study. setting A designated hospital for patients with covid-19 in Zhejiang province, China. ParticiPants 96 consecutively admitted patients with laboratory confirmed SARS-CoV-2 infection: 22 with mild disease and 74 with severe disease. Data were collected from 19

Mastocytosis is characterized by accumulations of mast cells in various organs (1). Most cases are indolent and confined to the skin, where discrete mast cell infiltrates are associated increased epidermal melanin, a clinical picture known as urticaria pigmentosa (UP). Other forms of mastocytosis combine UP with aggressive involvement of other organs or with haemotologic abnormalities (1-4). It is not known whether all forms of mastocytosis are true neoplasms or whether some might represent reactive hyperplasias (5-7). The c-KIT proto-oncogene encodes a type III receptor tyrosine kinase (KIT) that is critical to the development and survival of mast cells and melanocytes (8-11). The ligand for KIT (KL) can stimulate mast cell development, proliferation, and mediator release (9,12-17), as well as melanocyte proliferation and pigment production (18-20). To determine the role of c-KIT in the pathogenesis of mastocytosis, we examined tissue and cells isolated from a patient with UP and aggressive systemic mastocytosis with massive splenic involvement. We found a mutation that results in constitutive activation and expression of c-KIT in mast cells of both skin and spleen. This is the first in situ demonstration of an activation c-KIT mutation in neoplastic cells. It also demonstrates the clonal and neoplastic nature of this form of mastocytes.

The growth and differentiation of mast cells and melanocytes require stem cell factor (SCF), the ligand for the kit receptor tyrosine kinase. SCF may exist as a membrane-bound or soluble molecule. Abnormalities of the SCF-kit signaling pathway, with increased local concentrations of soluble SCF, have been implicated in the pathogenesis of the human disease cutaneous mastocytosis, but have not yet been shown to play a causal role. To investigate both the potential of SCF to cause mastocytosis and its role in epidermal melanocyte homeostasis, we targeted the expression of SCF to epidermal keratinocytes in mice with two different transgenes controlled by the human keratin 14 promoter. The transgenes contained cDNAs that either produced SCF, which can exist in both membrane-bound and soluble forms, or SCF, which remains essentially membrane bound. Murine epidermal keratinocyte expression of membrane-bound/ soluble SCF reproduced the phenotype of human cutaneous mastocytosis, with dermal mast cell infiltrates and epidermal hyperpigmentation, and caused the maintenance of a population of melanocytes in the interadnexal epidermis, an area where melanocytes and melanin are found in human skin but where they are not typically found in murine skin. Expression of membrane-bound SCF alone resulted in epidermal melanocytosis and melanin production, but did not by itself cause mastocytosis. We conclude, first, that a phenotype matching that of human mastocytosis can be produced in mice by keratinocyte overproduction of soluble SCF, suggesting a potential cause of this disease. Second, we conclude that keratinocyte expression of membrane-bound SCF results in the postnatal maintenance of epidermal melanocytes in mice. Since the resulting animals have skin that more closely approximates human skin than do normal mice, their study may be more relevant to human melanocyte biology than the study of skin of normal mice.

Interleukin 12 (IL-12), a heterodimeric cytokine composed of p40 and p35 chains, has potent immunologic effects in vitro. We used tuberculous pleuritis as a model to study the immunoregulatory potential of IL-12 in vivo at the site of human infectious disease. Messenger RNAs for p40 and p35 were detected in pleural fluid from six of six patients by reverse-transcription polymerase chain reaction. By using an ELISA that detected both free p40 and heterodimeric IL-12, we found that mean concentrations were 585±89 pg/ml in pleural fluid of patients with tuberculous pleuritis, which were significantly higher than those in serum of the same patients (54±36 pg/ ml), or in malignant pleural effusions (123±35 pg/ml). By using an ELISA specific for heterodimeric IL-12, we found that mean concentrations in pleural fluid of patients with tuberculous pleuritis were 165±28 pg/ml and undetectable in serum of the same patients, or in malignant pleural effusions. Clin. Invest. 1994. 93:1733-1739

Considerable mortality is still observed during the first years of life among patients with single ventricle. RV dominance is the most important risk factor for death but only before BCPS.

Clinical and immunologic evidence suggests that tuberculous pleuritis provides a model to understand protective immune mechanisms against Mycobacterium tuberculosis. We therefore evaluated the pattern of cytokine mRNA expression and cytokine production in pleural fluid and blood of patients with tuberculous pleuritis. RNA was extracted from mononuclear cells, reverse transcribed to cDNA, and amplified by polymerase chain reaction (PCR). After normalization for T-cell cDNA, cDNA from pleural fluid cells and peripheral blood mononuclear cells (PBMC) was amplified with cytokine-specific primers. PCR product was quantified by Southern blot. For the Thl cytokines gamma interferon (IFN-y) and interleukin-2 (IL-2), PCR product was greater in pleural fluid than in blood, whereas PCR product for the Th2 cytokine IL-4 was decreased in pleural fluid compared with blood. Concentrations of IFN-y were elevated in pleural fluid compared with serum, but IL-2, IL-4, and IL-5 were not detectable. Mean concentrations of IFN-y and IL-2 in supernatants of M. tuberculosis-stimulated pleural fluid cells were significantly greater than corresponding concentrations in supernatants of stimulated PBMC. In situ hybridization showed that increased IFN-y production by pleural fluid cells was associated with a 20to 60-fold increase in the frequency of antigen-reactive IFN-y-mRNA-expressing cells. Because IL-10 can be produced by T cells and macrophages, pleural fluid cells and PBMC were normalized for (-actin cDNA content and then amplified by PCR with IL-10-specific primers. IL-10 mRNA was greater in pleural fluid cells than in PBMC and was expressed predominantly by macrophages. IL-10 concentrations were elevated in pleural fluid versus serum. These data provide strong evidence for compartmentalization of Thl cytokines and IL-10 at the site of disease in humans with a resistant immune response to mycobacterial infection.

A tumor-on-a-chip platform with integration of decellularized liver matrix offers better biomimicry of tumor microenvironment and enhanced toxicity testing.

PurposeThe purpose of this study is to evaluate the dosimetric uncertainty associated with Gafchromic™ (EBT3) films and establish a practical and efficient film dosimetry protocol for Stereotactic Radiosurgery (SRS) and Stereotactic Body Radiotherapy (SBRT).Method and materialsEBT3 films were irradiated at each of seven different dose levels between 1 and 15 Gy with open fields and standard deviations of dose maps were calculated at each color channel for evaluation. A scanner non-uniform response correction map was built by registering and comparing film doses to the reference ion chamber array-based dose map delivered with the same doses. To determine the temporal dependence of EBT3 films, the average correction factors of different dose levels as a function of time were evaluated up to 4 days after irradiation.An integrated film dosimetry protocol was developed for dose calibration, calibration curve fitting, dose mapping, and profile/gamma analysis. Patient specific quality assurance (PSQA) was performed for 83 SRS/SBRT treatment plans, and analysis of the measurements and calculations are presented here.ResultsThe scanner response varied within 1 % for the field sizes less than 5 × 5 cm2, and up to 5 % for the field sizes of 10 × 10 cm2 for all color channels. The scanner correction method was able to remove visually evident, irregular detector responses for larger field sizes. The dose response of the film changed rapidly (~10 %) in the first two hours and became smooth plateaued afterwards, ~3 % change between 2 and 24 h. The uncertainties were approximately 1.5, 1.7 and 4.8 % over the dose range of 3~15 Gy for the red, green and blue channels. The green channel showed very high sensitivity and low uncertainty in the dose range between 10 and 15 Gy, which is suitable for SRS/SBRT commissioning and PSQA. The difference between the calculated dose and measured dose of ion chamber measurement at isocenter was −0.64 ± 2.02 for all plans, corresponding to a 95 % confidence interval of (−1.09, −0.26). The percentage of points passing the 3 %/1 mm gamma criteria in absolute dose, averaged over all tests was 95.0 ± 4.2.ConclusionWe have developed the EBT3 films based dosimetry protocol to obtain absolute dose values. The overall uncertainty has been established to be 1.5 % for SRS and SBRT PSQA.