Biohydrogen production and purification: Focusing on bioelectrochemical systems.

Innovative technologies on green hydrogen production become significant as the hydrogen economy has grown globally. Biohydrogen is one of green hydrogen production methods, and microbial electrochemical cells (MECs) can be key to biohydrogen provision. However, MECs are immature for biohydrogen technology due to several limitations including extracellular electron transfer (EET) engineering. Fundamental understanding of EET also needs more works to accelerate MEC commercialization. Interestingly, studies on biohydrogen gas purification are limited although biohydrogen gas mixture requires complex purification for use. To facilitate an MEC-based biohydrogen technology as the green hydrogen supply this review discussed EET kinetics, engineering of EET and direct interspecies electron transfer associated with hydrogen yield and the application of advanced molecular biology for improving EET kinetics. Finally, this article reviewed biohydrogen purification technologies to better understand purification and use appropriate for biohydrogen, focusing on membrane separation.

Regulatory Programs of B-cell Activation and Germinal Center Reaction Allow B-ALL Escape from CD19 CAR T-cell Therapy.

Chimeric antigen receptor (CAR) T-cell therapy has led to tremendous successes in the treatment of B-cell malignancies. However, a large fraction of treated patients relapse, often with disease expressing reduced levels of the target antigen. Here, we report that exposing CD19+ B-cell acute lymphoblastic leukemia (B-ALL) cells to CD19 CAR T cells reduced CD19 expression within hours. Initially, CD19 CAR T cells caused clustering of CD19 at the T cell-leukemia cell interface followed by CD19 internalization and decreased CD19 surface expression on the B-ALL cells. CD19 expression was then repressed by transcriptional rewiring. Using single-cell RNA sequencing and single-cell assay for transposase-accessible chromatin using sequencing, we demonstrated that a subset of refractory CD19low cells sustained decreased CD19 expression through transcriptional programs of physiologic B-cell activation and germinal center reaction. Inhibiting B-cell activation programs with the Bruton's tyrosine kinase inhibitor ibrutinib increased the cytotoxicity of CD19 CAR T cells without affecting CAR T-cell viability. These results demonstrate transcriptional plasticity as an underlying mechanism of escape from CAR T cells and highlight the importance of combining CAR T-cell therapy with targeted therapies that aim to overcome this plasticity. See related Spotlight by Zhao and Melenhorst, p. 1040.

Death Receptor 6 as a Prognostic Marker in Low-grade Glioma.

BACKGROUND/AIM: Low-grade gliomas (LGG) are heterogenous tumours, causing variable survivals in patients. Identifying molecular markers for a more accurate prognosis is, therefore, important. Since death receptor 6 (DR6) is up-regulated in gliomas and shows an aberrant signalling network, we tested its suitability as a prognostic marker. MATERIALS AND METHODS: DR6 was investigated in patient samples via PCR and western blot. Clinical data were analysed and compared to The Cancer Genome Atlas (TCGA) 'brain lower grade glioma' dataset. RESULTS: DR6 was found to be enhanced in LGG and its expression increased in recurrent LGG. The receptor showed a protective effect in primary LGG with a significantly elongated progression-free survival that was confirmed in the TCGA study. This effect was reversed in relapsed LGG in which cases with high DR6 expression reveal a shorter overall survival. CONCLUSION: DR6 is an interesting candidate for further studies regarding its effect as a prognostic marker, playing an opposing role in primary and relapsed LGG.

Effect of Low Frequency Cerebellar Repetitive Transcranial Magnetic Stimulation on Balance Impairment in Patients With Cerebral Infarction.

OBJECTIVE: To investigate the effect of low frequency cerebellar repetitive transcranial magnetic stimulation (rTMS) on balance impairment in patients with cerebral infarction. METHODS: Thirty-two patients were randomly divided into two groups: rTMS group (n=16) and control (n=16). In the rTMS group, treatment was performed five times per week for 2 weeks (10 sessions), and in the control group, a sham coil was used with the sound and sensation of scalp similar to the rTMS coil. Patients in both groups underwent a conventional rehabilitation program. Berg Balance Scale (BBS) was used as the primary outcome measurement. Timed Up and Go test (TUG), 10-m walk test (10mWT), and Activity-specific Balance Confidence scale (ABC) were used as the secondary outcome measurement. All scales were measured at baseline (T0), after 10 sessions of rTMS (T1), and at 4 weeks after treatment completion (T2) by therapists with over 5 years of clinical experience. RESULTS: There were significant improvements between T0 and T1, and between T0 and T2, for all assessed items in the rTMS group. Whereas there were significant improvements between T0 and T1, and between T0 and T2, for the BBS and 10mWT in the control group. TUG (-4.87±5.05 vs. -0.50±2.97 seconds) and ABC score (8.10±8.33 vs. 0.16±0.97) were observed significant differences in comparison of the changes from T0 to T1 between the two group. BBS score (4.40±3.66 vs. 1.88±3.14), TUG (-4.87±4.56 vs. -0.62±2.96 seconds) and ABC score (8.22±7.70 vs. -0.09±0.86) differed significantly from T0 to T2 between the two groups. CONCLUSION: Our findings suggest that low-frequency cerebellar rTMS is helpful for improving balance in patients with cerebral infarction, and maybe a beneficial treatment for these patients.

Single-Cell Profiling Reveals Metabolic Reprogramming as a Resistance Mechanism in BRAF-Mutated Multiple Myeloma.

PURPOSE: Although remarkably effective in some patients, precision medicine typically induces only transient responses despite initial absence of resistance-conferring mutations. Using BRAF-mutated myeloma as a model for resistance to precision medicine we investigated if BRAF-mutated cancer cells have the ability to ensure their survival by rapidly adapting to BRAF inhibitor treatment. EXPERIMENTAL DESIGN: Full-length single-cell RNA (scRNA) sequencing (scRNA-seq) was conducted on 3 patients with BRAF-mutated myeloma and 1 healthy donor. We sequenced 1,495 cells before, after 1 week, and at clinical relapse to BRAF/MEK inhibitor treatment. We developed an in vitro model of dabrafenib resistance using genetically homogeneous single-cell clones from two cell lines with established BRAF mutations (U266, DP6). Transcriptional and epigenetic adaptation in resistant cells were defined by RNA-seq and H3K27ac chromatin immunoprecipitation sequencing (ChIP-seq). Mitochondrial metabolism was characterized by metabolic flux analysis. RESULTS: Profiling by scRNA-seq revealed rapid cellular state changes in response to BRAF/MEK inhibition in patients with myeloma and cell lines. Transcriptional adaptation preceded detectable outgrowth of genetically discernible drug-resistant clones and was associated with widespread enhancer remodeling. As a dominant vulnerability, dependency on oxidative phosphorylation (OxPhos) was induced. In treated individuals, OxPhos was activated at the time of relapse and showed inverse correlation to MAPK activation. Metabolic flux analysis confirmed OxPhos as a preferential energetic resource of drug-persistent myeloma cells. CONCLUSIONS: This study demonstrates that cancer cells have the ability to rapidly adapt to precision treatments through transcriptional state changes, epigenetic adaptation, and metabolic rewiring, thus facilitating the development of refractory disease while simultaneously exposing novel vulnerabilities.

Nitro-Oleic Acid (NO2-OA) Improves Systolic Function in Dilated Cardiomyopathy by Attenuating Myocardial Fibrosis.

Nitro-oleic acid (NO2-OA), a nitric oxide (NO)- and nitrite (NO2-)-derived electrophilic fatty acid metabolite, displays anti-inflammatory and anti-fibrotic signaling actions and therapeutic benefit in murine models of ischemia-reperfusion, atrial fibrillation, and pulmonary hypertension. Muscle LIM protein-deficient mice (Mlp-/-) develop dilated cardiomyopathy (DCM), characterized by impaired left ventricular function and increased ventricular fibrosis at the age of 8 weeks. This study investigated the effects of NO2-OA on cardiac function in Mlp-/- mice both in vivo and in vitro. Mlp-/- mice were treated with NO2-OA or vehicle for 4 weeks via subcutaneous osmotic minipumps. Wildtype (WT) littermates treated with vehicle served as controls. Mlp-/- mice exhibited enhanced TGFß signalling, fibrosis and severely reduced left ventricular systolic function. NO2-OA treatment attenuated interstitial myocardial fibrosis and substantially improved left ventricular systolic function in Mlp-/- mice. In vitro studies of TGFß-stimulated primary cardiac fibroblasts further revealed that the anti-fibrotic effects of NO2-OA rely on its capability to attenuate fibroblast to myofibroblast transdifferentiation by inhibiting phosphorylation of TGFß downstream targets. In conclusion, we demonstrate a substantial therapeutic benefit of NO2-OA in a murine model of DCM, mediated by interfering with endogenously activated TGFß signaling.

Changes in Blood Glucose Level After Steroid Injection for Musculoskeletal Pain in Patients With Diabetes.

OBJECTIVE: To investigate changes in blood glucose level after steroid injection in patients with type 2 diabetes mellitus (DM) and factors affecting those changes. METHODS: We retrospectively studied 51 patients with type 2 DM who underwent steroid injection for shoulder and back pain. Mean fasting blood sugar (FBS) levels for 7 days before steroid injection was used as the baseline blood glucose level, which was compared with FBS levels for 14 days after steroid injection. We compared the differences in blood glucose changes between HbA1c >7% and HbA1c ≤7% groups and those between insulin and non-insulin treated groups. Demographic data, injection site, and steroid dose were analyzed. RESULTS: Compared to baseline, blood glucose significantly (p=0.012) elevated 1 day after steroid injection but not 2 days after injection. In the HbA1c >7% and insulin groups, blood glucose was significantly increased 1 day after injection compared to that in the HbA1c ≤7% (p=0.011) and non-insulin (p=0.024) groups, respectively. Higher HbA1c level before injection was significantly (p=0.003) associated with the degree of blood glucose increase 1 day after injection. No significant differences were noted in the degree of blood glucose increase according to injection site or steroid dose. CONCLUSION: Higher HbA1c level was associated with greater elevation in blood glucose 1 day after steroid injection. Careful monitoring of blood glucose is required on the first day after steroid injection in patients with poorly controlled DM.