A Phase I/II first-line study of R-CHOP plus B-cell receptor/NF-κB-double-targeting to molecularly assess therapy response

The ImbruVeRCHOP trial is an investigator-initiated, multicenter, single-arm, open label Phase I/II study for patients 61-80 years of age with newly diagnosed CD20+ diffuse large B-cell lymphoma and a higher risk profile (International Prognostic Index ≥2). Patients receive standard chemotherapy (CHOP) plus immunotherapy (Rituximab), a biological agent (the proteasome inhibitor Bortezomib) and a signaling inhibitor (the Bruton's Tyrosine Kinase-targeting therapeutic Ibrutinib). Using an all-comers approach, but subjecting patients to another lymphoma biopsy acutely under first-cycle immune-chemo drug exposure, ImbruVeRCHOP seeks to identify an unbiased molecular responder signature that marks diffuse large B-cell lymphoma patients at risk and likely to benefit from this regimen as a double, proximal and distal B-cell receptor/NF-κB-co-targeting extension of the current R-CHOP standard of care. EudraCT-Number: 2015-003429-32; ClinicalTrials.gov identifier: NCT03129828

A time-resolved proteomic and prognostic map of COVID-19.

COVID-19 is highly variable in its clinical presentation, ranging from asymptomatic infection to severe organ damage and death. We characterized the time-dependent progression of the disease in 139 COVID-19 inpatients by measuring 86 accredited diagnostic parameters, such as blood cell counts and enzyme activities, as well as untargeted plasma proteomes at 687 sampling points. We report an initial spike in a systemic inflammatory response, which is gradually alleviated and followed by a protein signature indicative of tissue repair, metabolic reconstitution, and immunomodulation. We identify prognostic marker signatures for devising risk-adapted treatment strategies and use machine learning to classify therapeutic needs. We show that the machine learning models based on the proteome are transferable to an independent cohort. Our study presents a map linking routinely used clinical diagnostic parameters to plasma proteomes and their dynamics in an infectious disease

A time-resolved proteomic and prognostic map of COVID-19

COVID-19 is highly variable in its clinical presentation, ranging from asymptomatic infection to severe organ damage and death. We characterized the time-dependent progression of the disease in 139 COVID-19 inpatients by measuring 86 accredited diagnostic parameters, such as blood cell counts and enzyme activities, as well as untargeted plasma proteomes at 687 sampling points. We report an initial spike in a systemic inflammatory response, which is gradually alleviated and followed by a protein signature indicative of tissue repair, metabolic reconstitution, and immunomodulation. We identify prognostic marker signatures for devising risk-adapted treatment strategies and use machine learning to classify therapeutic needs. We show that the machine learning models based on the proteome are transferable to an independent cohort. Our study presents a map linking routinely used clinical diagnostic parameters to plasma proteomes and their dynamics in an infectious disease

Clinical and virological characteristics of hospitalised COVID-19 patients in a German tertiary care centre during the first wave of the SARS-CoV-2 pandemic: a prospective observational study

Purpose: Adequate patient allocation is pivotal for optimal resource management in strained healthcare systems, and requires detailed knowledge of clinical and virological disease trajectories. The purpose of this work was to identify risk factors associated with need for invasive mechanical ventilation (IMV), to analyse viral kinetics in patients with and without IMV and to provide a comprehensive description of clinical course. Methods: A cohort of 168 hospitalised adult COVID-19 patients enrolled in a prospective observational study at a large European tertiary care centre was analysed. Results: Forty-four per cent (71/161) of patients required invasive mechanical ventilation (IMV). Shorter duration of symptoms before admission (aOR 1.22 per day less, 95% CI 1.10-1.37, p < 0.01) and history of hypertension (aOR 5.55, 95% CI 2.00-16.82, p < 0.01) were associated with need for IMV. Patients on IMV had higher maximal concentrations, slower decline rates, and longer shedding of SARS-CoV-2 than non-IMV patients (33 days, IQR 26-46.75, vs 18 days, IQR 16-46.75, respectively, p < 0.01). Median duration of hospitalisation was 9 days (IQR 6-15.5) for non-IMV and 49.5 days (IQR 36.8-82.5) for IMV patients. Conclusions: Our results indicate a short duration of symptoms before admission as a risk factor for severe disease that merits further investigation and different viral load kinetics in severely affected patients. Median duration of hospitalisation of IMV patients was longer than described for acute respiratory distress syndrome unrelated to COVID-19

Short-term azithromycin treatment promotes cornea allograft survival in the rat.

Any inflammatory response following corneal transplantation may induce rejection and irreversible graft failure. The purpose of this study is to analyze the anti-inflammatory effect of azithromycin (AZM) following experimental keratoplasty in rats.Corneal transplants were performed between Fisher-donor and Lewis-recipient rats. Recipients were postoperatively treated three times daily with AZM, miglyol, ofloxacin or dexamethasone eye drops. As an additional control, AZM was applied following syngeneic keratoplasty. Furthermore, short-term treatments with AZM for seven days perioperatively or with AZM only three days prior to the transplantation were compared to appropriate controls. All transplants were monitored clinically for opacity, edema, and vascularization. Infiltrating CD45(+), CD4(+), CD8(+), CD25(+), CD161(+) and CD163(+) cells were quantified via immunohistochemistry.AZM significantly promoted corneal graft survival compared with miglyol or ofloxacin treatment. This effect was comparable to topical dexamethasone. No adverse AZM effect was observed. Histology confirmed a significant reduction of infiltrating leukocytes. The short-term application of AZM for three days prior to transplantation or for seven days perioperatively reduced corneal graft rejection significantly compared with the controls.Along with antibiotic properties, topical AZM has a strong anti-inflammatory effect. Following keratoplasty, this effect is comparable to topical dexamethasone without the risk of steroid-induced adverse effects. Short-term treatment with AZM three days prior to the transplantation was sufficient to promote graft survival in the rat keratoplasty model. We therefore suggest further assessing the anti-inflammatory function of topical AZM following keratoplasty in humans

Clinical graft evaluation of AZM compared to miglyol.

<div><p>(A)Opacification, vascularization, and edema were monitored every three days by two independent investigators. A significant difference in opacification between the miglyol and AZM allogen group could be detected at day 3 (p < 0.05), day 9 (p < 0.01) and at day 12 (p < 0.001) after keratoplasty. No significant difference between AZM syngeneic (group 3) and allogeneic (group 2) could be detected. Vascularization was not influenced in all groups.</p> <p>(B)Vascularization was not affected by AZM compared to controls.</p> <p>(C)Corneal edema was reduced in the AZM allogen group compared to the miglyol group beginning at day 9 (p < 0.05). A representative photo on day 12 following keratoplasty is shown. Bars show group 4 (control, black) in comparison with group 2 (grey) and group 3 (control, light grey) with SEM.</p></div

Clinical Graft Assessment Score.

<p>Clinical Graft Assessment Score.</p

Immunohistological analysis of leukocytic infiltration.

<div><p>(A)Immunostainings on cryosections of miglyol-treated (group 4, black) and AZM-treated animals (group 2, light grey; group 3, dark grey) were performed on day 13. Infiltrating CD45⁺ leukocytes in group 2 were significantly reduced in comparison to group 4 (p<0.01). Red cells are positive for the antibody.</p> <p>(B)While no difference in the number of CD4⁺ T cells was seen in both groups, the infiltrating CD8⁺ T cells (p < 0.05) and CD25⁺ T cells (p < 0.05) were reduced significantly in the AZM-treated animals. </p> <p>(C)The number of infiltrating CD161⁺ NK cells (p < 0.05) and of CD163⁺ macrophages (p < 0.05) in the central corneal stroma was significant reduced.</p></div

Graft survival.

<div><p>(A)Clear graft survival following keratoplasty is shown by Kaplan-Meier estimator. Treatment with miglyol (group 4) led to rejection at day 13 [12.5–13.5], whereas with AZM (group 3), no rejection was detected. When treating the allogeneic transplanted animals with AZM (group 2), rejection occurred at day 28.5 [20.3–36.6] postoperatively. AZM allogeneic compared with the control with miglyol did promote graft survival (p < 0.004). </p> <p>(B) Treatment with ofloxacin (group 5) led to rejection at day 13, similar to miglyol, while AZM improved graft survival significantly. </p></div

Clinical graft evaluation of azithromycin and dexamethasone following keratoplasty.

<div><p>(A)Orthotopic penetrating keratoplasties between Lewis and Fisher rats were performed. Total graft survival post keratoplasty is shown by Kaplan-Meier curves. Rejection was defined as total graft opacification (opacity grade 4). With AZM (group 2), rejection occurred at day 28.5 [20.3–36.6] postoperatively. No rejection occurred in the DM-treated group (group 1) (p < 0.14). </p> <p>(B)Immunostainings on cryosections of DM-treated (control, black) and AZM-treated animals (light grey). Total leukocytic infiltrate was counted in three different squares per animal by two independent investigators. No significant difference of total leukocytic infiltrate (red cells) between the group 1 and 2 could be detected at day 35. Bars show group 1 (control, black) in comparison to group 2 (grey) with SEM.</p></div