Autologous antigen-presenting cells efficiently expand piggyBac transposon CAR-T cells with predominant memory phenotype.

The quality of chimeric antigen receptor (CAR)-T cell products, including the expression of memory and exhaustion markers, has been shown to influence their long-term functionality. The manufacturing process of CAR-T cells should be optimized to prevent early T cell exhaustion during expansion. Activation of T cells by monoclonal antibodies is a critical step for T cell expansion, which may sometimes induce excess stimulation and exhaustion of T cells. Given that piggyBac transposon (PB)-based gene transfer could circumvent the conventional pre-activation of T cells, we established a manufacturing method of PB-mediated HER2-specific CAR-T cells (PB-HER2-CAR-T cells) that maintains their memory phenotype without early T cell exhaustion. Through stimulation of CAR-transduced T cells with autologous peripheral blood mononuclear cell-derived feeder cells expressing both truncated HER2, CD80, and 4-1BBL proteins, we could effectively propagate memory-rich, PD-1-negative PB-HER2-CAR-T cells. PB-HER2-CAR-T cells demonstrated sustained antitumor efficacy in vitro and debulked the HER2-positive tumors in vivo. Mice treated with PB-HER2-CAR-T cells rejected the second tumor establishment owing to the in vivo expansion of PB-HER2-CAR-T cells. Our simple and effective manufacturing process using PB system and genetically modified donor-derived feeder cells is a promising strategy for the use of PB-CAR-T cell therapy.

Takayasu Arteritis With Antiphosphatidylserine/Prothrombin Antibody-Positive Antiphospholipid Syndrome: Case Report and Literature Review.

A relationship between Takayasu arteritis (TA) and positive antiphospholipid antibody states has been pointed out, but patients with TA complicated with antiphospholipid antibody syndrome (APS) are rare. Here we report the case of a 17-year-old Japanese man diagnosed with TA based on pulselessness of the left brachial artery, discrepancy of blood pressure between the upper extremities, and arterial wall thickening and narrowing of artery in contrast computed tomography. He was also diagnosed with provisional APS based on a pulmonary infarction without narrowing of the pulmonary artery and positive antiphosphatidylserine/prothrombin antibody. The patient also had concurrent Crohn's disease (CD) based on histopathological findings, which may have been associated with TA. We started high-dose corticosteroid therapy and anticoagulation therapy, and his symptoms including fever, dizziness, chest pain, and lower-right uncomfortable abdomen improved.We reviewed 9 cases of TA with APS including our patient by conducting a PubMed search. Based on past reports, we considered the relationship among TA, APS, and CD.Clinicians should bear in mind that many etiologies can exist in 1 patient, and differential diagnoses are essential.

No Dynamic Changes in Inflammation-related Microcirculatory Parameters in Developing Rats During Local Cortex Exposure to Microwaves.

The biological effects of exposing the developing brain to radiofrequency electromagnetic fields (RF) are still unclear. Our experiments investigated whether three inflammation-related, microcirculatory parameters in juvenile and young adult rats were modified during local cortex exposure to RF under non-thermal conditions. The cortex tissue was locally exposed to 1457 MHz RF at an averaged specific absorption rate of 2.0 W/kg in the target area for 50 min and variations of pial venule parameter were measured directly in vivo. There was no significant difference in hemodynamics, plasma velocity or vessel diameter, between exposed and sham-exposed groups for either rat development stage. No increase related to RF exposure was found in leukocyte adhesion to endothelial cells in any microvessels observed. These findings suggest that RF is unlikely to initiate inflammatory responses, at least under these exposure conditions.

No Dynamic Changes in Blood-brain Barrier Permeability Occur in Developing Rats During Local Cortex Exposure to Microwaves.

Little information is available about the effects of exposure to radiofrequency electromagnetic fields (RF) on cerebral microcirculation during rat developmental stages. We investigated whether the permeability of the blood-brain barrier (BBB) in juvenile and young adult rats was modified during local cortex exposure to RF under non-thermal conditions. The cortex tissue targeted was locally exposed to 1457 MHz RF at an average specific absorption rate of 2.0 W/kg in the target area for 50 min and permeability changes in the BBB of the pia mater were measured directly, using intravital fluorescence microscopy. There was no significant difference in extravasation of intravenously-injected dye between exposed and sham-exposed groups of either category of rats. No histological evidence of albumin leakage was found in any of the brains just after exposure, indicating that no traces of BBB disruption remained. These findings suggest that no dynamic changes occurred in BBB permeability of the rats at either of these developmental stages, even during local RF exposure at non-thermal levels.

No changes in cerebral microcirculatory parameters in rat during local cortex exposure to microwaves.

The aim of this study was to determine whether cerebral microcirculatory parameters in rats were modified during local cortex exposure to a radiofrequency electromagnetic field (RF) under non-thermal conditions. The cortex tissue targeted was locally exposed to 1439 MHz RF using a figure-8 loop antenna at an averaged specific absorption rate of 2.0 W/kg in the target area for 50 min. Three microcirculatory parameters related to cerebral inflammation were measured by the cranial window method in real-time under RF exposure. No extravasation of intravenously injected fluorescent dye was observed during RF exposure. There was no significant difference either in pial venule blood flow velocity or diameter between exposed and sham-exposed rats. Histological evaluation for the brain immediately after RF exposure did not reveal any serum albumin leakage sites or degenerate neurons. These findings suggest that no dynamic changes occurred in cerebral microcirculation even during local cortex exposure under these conditions.

Effects of short-term W-CDMA mobile phone base station exposure on women with or without mobile phone related symptoms.

To investigate possible health effects of mobile phone use, we conducted a double-blind, cross-over provocation study to confirm whether subjects with mobile phone related symptoms (MPRS) are more susceptible than control subjects to the effect of electromagnetic fields (EMF) emitted from base stations. We sent questionnaires to 5,000 women and obtained 2,472 valid responses from possible candidates; from these, we recruited 11 subjects with MPRS and 43 controls. There were four EMF exposure conditions, each of which lasted 30 min: continuous, intermittent, and sham exposure with and without noise. Subjects were exposed to EMF of 2.14 GHz, 10 V/m (W-CDMA), in a shielded room to simulate whole-body exposure to EMF from base stations, although the exposure strength we used was higher than that commonly received from base stations. We measured several psychological and cognitive parameters pre- and post-exposure, and monitored autonomic functions. Subjects were asked to report on their perception of EMF and level of discomfort during the experiment. The MPRS group did not differ from the controls in their ability to detect exposure to EMF; nevertheless they consistently experienced more discomfort, regardless of whether or not they were actually exposed to EMF, and despite the lack of significant changes in their autonomic functions. Thus, the two groups did not differ in their responses to real or sham EMF exposure according to any psychological, cognitive or autonomic assessment. In conclusion, we found no evidence of any causal link between hypersensitivity symptoms and exposure to EMF from base stations.

Effects of acute exposure to a 1439 MHz electromagnetic field on the microcirculatory parameters in rat brain.

UNLABELLED: THE AIM of this study was to determine the potential of radio-frequency electromagnetic fields (RF-EMF) to affect cerebral microcirculation, including blood-brain barrier function, in rat brain. MATERIALS AND METHODS: The head of the rat was exposed for 10 min to 1439 MHz RF-EMF having three intensity doses: 0.6, 2.4, 4.8 W/kg of brain averaged specific absorption rate (BASAR). Four microcirculatory parameters: blood-brain barrier permeability, leukocyte behavior, plasma velocity, and vessel diameter were measured before and after RF-EMF exposure using a closed cranial window method. RESULTS: No extravasation of intravenously injected dyes from pial venules was found at any BASAR level. No significant changes in the number of endothelial-adhering leukocytes after exposure were found. The hemodynamics indicated that the plasma velocities and vessel diameters remained constant within the physiological range throughout each exposure. CONCLUSION: These findings suggest that there were no effects on the cerebral microcirculation under the given RF-EMF exposure conditions.

Effects of subchronic exposure to a 1439 MHz electromagnetic field on the microcirculatory parameters in rat brain.

UNLABELLED: THE AIM of this study was to investigate whether repeated exposure to radio frequency electromagnetic field (RF-EMF) of 1439 MHz affects the cerebral microcirculation, including blood-brain barrier function, in a rat brain. MATERIALS AND METHODS: The head of the rat was exposed for four weeks (60 min/day, 5 days/week) to RF-EMF at 2.4 W/kg of brain averaged specific absorption rate (BASAR). Three microcirculatory parameters: blood-brain barrier permeability, leukocyte behavior and plasma velocity were measured before and after RF-EMF exposure using a closed cranial window method. RESULTS: No extravasation of intravenously injected dyes from pial venules was found at any BASAR level. No significant changes in the number of endothelial-adhering leukocytes after exposure were found. The plasma velocity remained constant within the physiological range through each exposure. CONCLUSION: These findings suggest that there were no effects on the cerebral microcirculation under the given RF-EMF exposure conditions.

Long-term observation of pial microcirculatory parameters using an implanted cranial window method in the rat.

The aim of this study was to confirm whether our improved closed cranial window (CCW) method could be used for long-term microscopical observation of pial microcirculation intravitally in the rat. We investigated chronological changes in three microcirculatory parameters: permeability of blood-brain barrier, leukocyte behavior, and plasma velocities in the pial venules, immediately after implantation (control group) and at one and four weeks after implantation in different age-matched rats (implanted group). No extravasation of sodium fluorescein from pial venules was confirmed in any observation period. The number of endothelial-adhering leukocytes in the implanted group kept within the physiological range, being similar to those of the control group. The velocities of fluorescent microspheres flowing in pial venules showed no noticeable changes between the two groups. These findings suggest that our CCW method allows long-term observation of the pial microcirculation without any pathophysiological changes in the evaluated parameters up to four weeks after the implantation.