Interleukin-7 improves the fitness of regulatory T cells for adoptive transfer.

Adoptive regulatory T-cell (Treg) transfer has emerged as a promising therapeutic strategy for regulating immune responses in organ transplantation, graft versus host disease, and autoimmunity, including Type 1 diabetes. Traditionally, Treg for adoptive therapy have been sorted and expanded in vitro using high doses of IL-2, demonstrating stability and suppressive capabilities. However, limitations in their long-term survival post-infusion into patients have been observed. To address this challenge, we investigated a novel expansion protocol incorporating interleukin-7 (IL-7) alongside the traditional method utilizing IL-2 (referred to as IL-7 method, IL-7M). Our study revealed that naïve Treg express significant levels of CD127 and display robust responsiveness to IL-7, characterized by STAT-5 phosphorylation. Expanding naïve Treg with the IL-7M protocol led to a substantial enrichment of CD45RA+ CD62L+ CD95+ Treg but showing a reduction in the final cell yield and suppressive function. Moreover, Treg expanded with the IL-7M exhibited preserved telomere length and demonstrated enhanced resistance to cytokine withdrawal and fas-mediated apoptosis. When transferred into NSG mice IL-7M-Treg persisted longer and reduced the expansion of T cells, but did not significantly reduce the severity of xenoGvHD. In conclusion, our data demonstrate the feasibility of expanding naïve Treg in the presence of IL-7 to generate a Treg product enriched in poorly differentiated CD45RA+ cells with enhanced survival capabilities.

Impact of Assistant Experience on Perioperative Outcomes of Simple and Radical Laparoscopic Nephrectomy: Does It Really Matter?

Background and Objectives: While systematic reviews highlight the advantages of laparoscopic nephrectomy over traditional open surgery, the impact of an assistant's experience on surgical outcomes remains unclear. This study aims to evaluate whether the level of assistant expertise influences laparoscopic nephrectomy outcomes. Materials and Methods: Retrospective data from our institutional database were analyzed for patients who underwent laparoscopic nephrectomy between January 2018 and December 2022. Procedures were performed by a highly experienced surgeon, including postgraduate year (PGY)-3 to PGY-5 residents as assistants. Senior-level assistants had completed at least 10 procedures. Patient characteristics, surgical outcomes, and postoperative details were collected. Multivariable linear and logistic regression models were performed to test the effect of assistant experience (low vs. high) on estimated blood loss (EBL), length of stay (LOS), operative time (OT), and postoperative complications. Results: 105 patients were included, where 53% had highly experienced assistants and 47% had less experienced ones. Low assistant experience and higher BMI predicted longer operative time (OT), confirmed by multivariable regression (ß = 40.5, confidence interval [CI] 18.7-62.3, p < 0.001). Assistant experience did not significantly affect EBL or LOS after adjusting for covariates (ß = -14.2, CI -91.8-63.3, p = 0.7 and ß = -0.83, CI -2.7-1.02, p = 0.4, respectively). There was no correlation between assistant experience and postoperative complications. Conclusions: Assistant experience does not significantly impact complications, EBL, and LOS in laparoscopic nephrectomy. Surgeries with less experienced assistants had longer OT, but the overall clinical impact seems limited. Trainee involvement remains safe, guided by experienced surgeons.

Nitrogen-15 dynamic nuclear polarization of nicotinamide derivatives in biocompatible solutions.

Dissolution dynamic nuclear polarization (dDNP) increases the sensitivity of magnetic resonance imaging by more than 10,000 times, enabling in vivo metabolic imaging to be performed noninvasively in real time. Here, we are developing a group of dDNP polarized tracers based on nicotinamide (NAM). We synthesized 1-15N-NAM and 1-15N nicotinic acid and hyperpolarized them with dDNP, reaching (13.0 ± 1.9)% 15N polarization. We found that the lifetime of hyperpolarized 1-15N-NAM is strongly field- and pH-dependent, with T1 being as long as 41 s at a pH of 12 and 1 T while as short as a few seconds at neutral pH and fields below 1 T. The remarkably short 1-15N lifetime at low magnetic fields and neutral pH drove us to establish a unique pH neutralization procedure. Using 15N dDNP and an inexpensive rodent imaging probe designed in-house, we acquired a 15N MRI of 1-15N-NAM (previously hyperpolarized for more than an hour) in less than 1 s.

Performance and reproducibility of 13C and 15N hyperpolarization using a cryogen-free DNP polarizer.

The setup, operational procedures and performance of a cryogen-free device for producing hyperpolarized contrast agents using dissolution dynamic nuclear polarization (dDNP) in a preclinical imaging center is described. The polarization was optimized using the solid-state, DNP-enhanced NMR signal to calibrate the sample position, microwave and NMR frequency and power and flip angle. The polarization of a standard formulation to yield ~ 4 mL, 60 mM 1-13C-pyruvic acid in an aqueous solution was quantified in five experiments to P(13C) = (38 ± 6) % (19 ± 1) s after dissolution. The mono-exponential time constant of the build-up of the solid-state polarization was quantified to (1032 ± 22) s. We achieved a duty cycle of 1.5 h that includes sample loading, monitoring the polarization build-up, dissolution and preparation for the next run. After injection of the contrast agent in vivo, pyruvate, pyruvate hydrate, lactate, and alanine were observed, by measuring metabolite maps. Based on this work sequence, hyperpolarized 15N urea was obtained (P(15N) = (5.6 ± 0.8) % (30 ± 3) s after dissolution).

From women to women-hematuria during therapy for metastatic breast cancer, what to suspect and when to be alarmed; Bladder metastasis from breast cancer-our experience and a systematic literature review.

Breast cancer is one of the most important causes of premature mortality in women worldwide. Around 12% of breast cancer patients will develop metastatic disease, a stage associated with poor prognosis, and only 26% of patients are likely to survive for at least 5 years after being diagnosed. Although the most common sites where breast cancer tends to spread are bones, lungs, brain, and liver, it is important that physicians consider other less frequent organs and viscera, like the bladder, as a target destination. In this article we report our experience with this rare form of metastases and a systematic literature review. We analyzed case reports, case series, and review articles present in PubMED/MEDLINE up to March 2022. We excluded the nonrelevant articles, editorials, letters to the editor, and articles written in other languages. We identified a total of 302 articles, with 200 articles being removed before screening; therefore, the total number of abstracts reviewed was 102. Fifty-five articles were excluded before full text review because they did not meet the inclusion criteria, and one article was not retrievable. Therefore, we included a total of 45 articles in this review. The intention of this review is to highlight the importance of the early detection of bladder metastases and to facilitate the diagnostic process for the responsible physician. The most common signs and symptoms and breast cancer subtype associated with bladder metastases, as well as overall survival after their detection, were all assessed. Bladder metastases from metastatic breast cancer are prevalent in the invasive, lobular breast cancer subtype; most patients present with hematuria (39.5%) and the relative 5-year survival rate is 2%. The main limitations of this review are the low number of cases reported in the literature, clinical and pathological differences between the individual cases, and absence of the control group. This study was not funded.

[Hyperpolarized 13C magnetic resonance imaging-a window into metabolism : High-resolution images of human metabolism]. / Hyperpolarisierte 13C­Magnetresonanztomographie ­ ein Fenster in den Stoffwechsel : Hochauflösende Darstellung des humanen Metabolismus.

CLINICAL ISSUE: Despite being one of the main pillars of modern diagnostics, magnetic resonance imaging (MRI) uses only a tiny fraction of its potential: no more than a millionth of all nuclear spins contribute to the MRI signal. In order to increase this fraction, called polarization, MRI scanners with stronger magnetic fields are being developed. However, even the most modern scanners do not exploit the potential of MRI. METHODOLOGICAL INNOVATIONS: To make full use of this potential, hyperpolarized MRI (HP-MRI) is an excellent tool: quantum mechanical tricks can be used to generate contrast agents whose nuclear spins can deliver a MRI signal that is up to a 100,000 times stronger. This signal enhancement allows imaging of in vivo processes that would be otherwise impossible to measure. It is particularly interesting to introduce these magnetically labeled nuclei into metabolic processes so that the metabolism can be investigated non-invasively and in vivo. PERFORMANCE: Small but diagnostically important changes in metabolism could be found before macroscopic tissue changes were otherwise visible. High-resolution images can be acquired within a few 100 ms, enabling metabolic monitoring in real-time. Heart, brain, and prostate are among the organs that have already been investigated in over 90 clinical trials using this emerging technology. ACHIEVEMENTS: So far, displaying tissue in a similar manner was only possible using nuclear medicine, e.g., positron emission tomography (PET) utilizing radionuclides and without resolution of various metabolic steps. A change in tumor metabolism following treatment was shown within hours in HP-MRI. These applications coupled with background information about the technology are the subject of this review.

Imaging Inflammation - From Whole Body Imaging to Cellular Resolution.

Imaging techniques have evolved impressively lately, allowing whole new concepts like multimodal imaging, personal medicine, theranostic therapies, and molecular imaging to increase general awareness of possiblities of imaging to medicine field. Here, we have collected the selected (3D) imaging modalities and evaluated the recent findings on preclinical and clinical inflammation imaging. The focus has been on the feasibility of imaging to aid in inflammation precision medicine, and the key challenges and opportunities of the imaging modalities are presented. Some examples of the current usage in clinics/close to clinics have been brought out as an example. This review evaluates the future prospects of the imaging technologies for clinical applications in precision medicine from the pre-clinical development point of view.

Digital chest tomosynthesis: the 2017 updated review of an emerging application.

Lung cancer is the leading cause of cancer death and second most common cancer among both men and women, but most of them are detected when patients become symptomatic and in late-stage. Chest radiography (CR) is a basic technique for the investigation of lung cancer and has the benefit of convenience and low radiation dose, but detection of malignancy is often difficult. The introduction of computed tomography (CT) for screening has increased the proportion of lung cancer detected but with higher exposure dose and higher costs. Digital chest tomosynthesis (DCT), a tomographic technique, may offer an alternative to CT. DCT uses a conventional radiograph tube, a flat-panel detector, a computer-controlled tube mover and reconstruction algorithms to produce section images. It shows promise in the detection of potentially malignant lung nodules, with higher sensibility than CR, and is emerging as a low-dose and low-cost alternative to CT to improve treatment decisions. In fact, an increasing number of researchers are showing that tomosynthesis could have a role in the detection of lung cancer, in addition to its present role in breast screening. However, DCT offers some limitations, such as limited depth resolution, which may explain the difficulty in detecting pathologies in the subpleural region and the occurrence of artefacts from medical devices. Once solved these limitations and once more studies supporting its use will be available, DCT could become the first-line lung cancer screening tool among patients at considerable risk of lung cancer.

Genetically modified laboratory animals in the name of the 3Rs?

Although the introduction of GM animal models at first was viewed as a potent way to enhance experimental biology in the name of the 3Rs by its proponents, over the years, the number of animals used has greatly increased and concerns about the suffering of these animals have emerged in the debate. The purpose of this contribution is to show the need and the urgency for a systematic evaluation of genetically modified laboratory animals (GM animals) according to the 3Rs principle. This evaluation presents various difficulties due to the special features of the genetic modifications of animals, the variety of scientific purposes connected with the use of these animals, the lack of coherent statistical data about this use and the difficulties related to the welfare assessment of these animals. In this article I discuss the significance of the procedures involving GM animals for each of the 3R principles. On this basis, I offer an answer to the question of whether these procedures are compatible with the spirit of the 3Rs.