Experimental Search for Invisible Dark Matter Axions around 22 µeV.

The axion has emerged as the most attractive solution to two fundamental questions in modern physics related to the charge-parity invariance in strong interactions and the invisible matter component of our Universe. Over the past decade, there have been many theoretical efforts to constrain the axion mass based on various cosmological assumptions. Interestingly, different approaches from independent groups produce good overlap between 20 and 30 µeV. We performed an experimental search to probe the presence of dark matter axions within this particular mass region. The experiment utilized a multicell cavity haloscope embedded in a 12 T magnetic field to seek for microwave signals induced by the axion-photon coupling. The results ruled out the KSVZ axions as dark matter over a mass range between 21.86 and 22.00 µeV at a 90% confidence level. This represents a sensitive experimental search guided by specific theoretical predictions.

Effect of depth of discharge (DOD) on cycling in situ formed Li anodes.

Lithium-metal solid-state batteries (LMSSBs) have garnered immense interest due to their potential to enhance safety and energy density compared to traditional Li-ion batteries. The anode-free approach to manufacturing Li-metal anodes could provide the additional benefit of reducing cost. However, a lack of understanding of the mechano-electrochemical behavior related to the cycling of in situ formed Li anodes remains a significant challenge. To bridge this knowledge gap, this work aims to understand the cycling behavior of in situ formed Li anodes on garnet Li7La3Zr2O12 (LLZO) solid-electrolyte as a function of the depth of discharge (DOD). The results of this study show that cycling in situ formed Li of 3 mA h cm-2 with a DOD of 66% leads to unstable cycling, while cycling with a DOD of 33% exhibits stable cycling. Furthermore, we observed interfacial deterioration and inhomogeneity of in situ formed Li anodes during cycling with a DOD of 66%. This study provides mechanistic insight into the factors that affect stable cycling that can help guide approaches to improve the cycling behavior of in situ formed Li anodes.

Optimizing and determining the click chemistry mediated Cu-64 radiolabeling and physiochemical characteristics of trastuzumab conjugates.

Over the last decade, 64Cu-labeling of monoclonal antibody (mAb) via inverse electron demand Diels-Alder click chemistry (IEDDA) have received much attention. Despite the tetrazine-transcyclooctene (Tz-TCO) click chemistry's convenience and efficiency in mAb labeling, there is limited information about the ideal parameters in the development of click chemistry mediated (radio)immunoconjugates. This encourages us to conduct a systematic optimization while concurrently determining the physiochemical characteristics of the model mAb, trastuzumab, and TCO conjugates. To accomplish this, we investigated a few critical parameters, first, we determined the degree of conjugations with varying molar equivalents (eq.) of TCO (3, 5, 10, and 15 eq.). Through analytical techniques like size exclusion chromatography, dynamic light scattering, and zeta potential, qualitative analysis were performed to determine the purity, degree of aggregation and net charge of the conjugates. We found that as the degree of conjugation increased the purity of intact mAb fraction is compromised and net charge of conjugates became less positive. Next, all trastuzumab-PEG4-TCO conjugates with varying molar ratio and quantity (30, 50, 100, 200, 250 µg) were radiolabeled with 64Cu-NOTA-PEG4-Tz via IEDDA click chemistry and radiochemical yields were determined by radio-thin layer chromatography. The radiochemical yields of trastuzumab conjugates improved with increased amount and molar ratio. Next, we investigated the effect of the radioprotectant ascorbic acid (AA) of varied concentrations (0.25, 0.5, 0.75, 1 mM) on radiochemical yields and subsequent pharmacokinetics. A concentration of 0.25 mM of AA was found to be optimal for click reaction and in vivo biodistribution. Finally, we investigated the indirect influence of bioconjugation buffers on radiochemical yields and biodistribution in NIH3T6.7 tumor models that resulted approximately ∼11 %ID/g tumor uptake.

In vivo detection of hydrogen sulfide in the brain of live mouse: application in neuroinflammation models.

PURPOSE: Hydrogen sulfide (H2S) plays important roles in brain pathophysiology. However, nuclear imaging probes for the in vivo detection of brain H2S in living animals have not been developed. Here, we report the first nuclear imaging probe that enables in vivo imaging of endogenous H2S in the brain of live mice. METHODS: Utilizing a bis(thiosemicarbazone) backbone, a fluorescent ATSM-FITC conjugate was synthesized. Its copper complex, Cu(ATSM-FITC) was thoroughly tested as a biosensor for H2S. The same ATSM-FITC ligand was quantitatively labeled with [64Cu]CuCl2 to obtain a radioactive [64Cu][Cu(ATSM-FITC)] imaging probe. Biodistribution and positron emission tomography (PET) imaging studies were performed in healthy mice and neuroinflammation models. RESULTS: The Cu(ATSM-FITC) complex reacts instantly with H2S to release CuS and becomes fluorescent. It showed excellent reactivity, sensitivity, and selectivity to H2S. Endogenous H2S levels in living cells were successfully detected by fluorescence microscopy. Exceptionally high brain uptake of [64Cu][Cu(ATSM-FITC)] (> 9% ID/g) was observed in biodistribution and PET imaging studies. Subtle changes in brain H2S concentrations in live mice were accurately detected by quantitative PET imaging. Due to its dual modality feature, increased H2S levels in neuroinflammation models were characterized at the subcellular level by fluorescence imaging and at the whole-body scale by PET imaging. CONCLUSION: Our biosensor can be readily utilized to study brain H2S function in live animal models and shows great potential as a novel imaging agent for diagnosing brain diseases.

Vertical stacking of three-dimensional nanostructures via an aerosol lithography for advanced optical applications.

In this report, we introduce a method utilizing ion-assisted aerosol lithography to stack 3D nanostructures vertically. The stacked 3D nanostructures exhibit extraordinary optical properties: the double layer 3D nanostructures show more than 5-fold increased surface enhanced Raman scattering intensities compared to their single layer counterpart. This unusual enhancement of Raman intensity implies the existence of additional vertical hotspots formed by interlayer cavity effects between the lower and upper nanostructures. Allowing for full three-dimensional control in nanofabrication, this work provides a reliable way to create complex-shaped advanced optical nanostructures with non-intuitive bulk optical properties.

Fermented Whey Protein Supplementation Improves Muscular Strength, Muscle Parameters, and Physical Performance in Middle-Aged Korean Adults: An 8-Week Double Blind Randomized Controlled Trial.

The present study evaluated the effects of fermented whey protein using kimchi lactic acid bacteria Lactobacillus casei DK211 on skeletal muscle mass, muscle strength, and physical performance in healthy middle-aged males performing regular resistance exercises. Effective protein supplementation and regular exercise are two important factors for improving muscle health. Therefore, in this study, the effects of consuming fermented whey protein twice a day were investigated and compared with that of non-fermented supplementation. Forty-eight males (average age 44.8) were randomly assigned to two groups: Fermented whey protein supplementation (FWPS) and non-fermented whey protein concentration supplementation (WPCS) groups. Each group ingested 37 g of FWPS or WPCS twice a day for eight weeks. Body composition, muscle strength, and physical performance were assessed pre- and post-intervention. Independent t-tests or chi-square tests for the categorical variables were performed for analyzing the observations. FWPS was effective in promoting the physical performance in dynamic balance measurement and muscle health, indicated through the increment in grip strength (left), upper arm circumference, and flat leg circumference from the baseline. However, similar improvements were not observed in the WPCS group. These results imply that whey protein fermented by L. casei DK211 is an effective protein supplement for enhancing muscle health in males performing regular resistance exercises.

Preclinical Evaluation of hnRNPA2B1 Antibody in Human Triple-Negative Breast Cancer MDA-MB-231 Cells via PET Imaging.

Triple-negative breast cancer (TNBC) does not express estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. Because TNBC lacks the expression of commonly targeted receptors, it is challenging to develop a new imaging agent for this cancer subtype. Heterogeneous nuclear ribonucleoproteins (hnRNPs) are RNA-protein complexes that have been linked to tumor development and progression. Considering the high expression of hnRNPA2B1, an hnRNP subtype, in TNBC MDA-MB-231 cells, this study aimed to develop a novel hnRNPA2B1 antibody-based nuclear imaging agent. The hnRNPA2B1-specific antibody was radiolabeled with 64Cu and evaluated in vitro and in vivo. The trans-cyclooctene (TCO) was functionalized on the antibody to obtain hnRNP-PEG4-TCO and reactive tetrazine (Tz) on the ultrastable bifunctional chelator PCB-TE2A-alkyne to yield PCB-TE2A-Tz for the inverse electron demand Diels-Alder reaction. The 64Cu-radiolabeled antibody was administered and imaged at 1-18 h time points for conventional imaging. Alternatively, the unlabeled antibody conjugate was administered, and 48 h later radiolabeled 64Cu-PCB-TE2A-Tz was administered to the same mice for the pretargeting strategy and imaged at the same time intervals for direct comparison. The tumor was successfully visualized in both strategies, and comparatively, pretargeting showed superior results. The 64Cu-PCB-TE2A-Tz was successfully clicked at the tumor site with hnRNP-PEG4-TCO and the non-clicked were concurrently eliminated. This led to increase the tumor uptake with extremely high tumor-to-background ratio manifested by positron emission tomography (PET) imaging and biodistribution studies.

Successful Application of CuAAC Click Reaction in Constructing 64Cu-Labeled Antibody Conjugates for Immuno-PET Imaging.

Immuno-positron emission tomography (immuno-PET) is a rapidly growing imaging technique in which antibodies are radiolabeled to monitor their in vivo behavior in real time. However, effecting the controlled conjugation of a chelate-bearing radioactive atom to a bulky antibody without affecting its immunoreactivity at a specific site is always challenging. The in vivo stability of the radiolabeled chelate is also a key issue for successful tumor imaging. To address these points, a facile ultra-stable radiolabeling platform is developed by using the propylene cross-bridged chelator (PCB-TE2A-alkyne), which can be instantly functionalized with various groups via the click reaction, thus enabling specific conjugation with antibodies as per choice. The PCB-TE2A-tetrazine derivative is selected to demonstrate the proposed strategy. The antibody trastuzumab is functionalized with the trans-cyclooctene (TCO) moiety in the presence or absence of the PEG linker. The complementary 64Cu-PCB-TE2A-tetrazine is synthesized via the click reaction and radiolabeled with 64Cu ions, which then reacts with the aforementioned TCO-modified antibody via a rapid biorthogonal ligation. The 64Cu-PCB-TE2A-trastuzumab conjugate is shown to exhibit excellent in vivo stability and to maintain a higher binding affinity toward HER2-positive cells. The tumor targeting feasibility of the radiolabeled antibody is evaluated in tumor models. Both 64Cu-PCB-TE2A-trastuzumab conjugates show high tumor uptakes in biodistribution studies and enable unambiguous tumor visualization with minimum background noise in PET imaging. Interestingly, the 64Cu-PCB-TE2A-PEG4-trastuzumab containing an additional PEG linker displays a much faster body clearance compared to its counterpart with less PEG linker, thus affording vivid tumor imaging with an unprecedentedly high tumor-to-background ratio.

A short PEG linker alters the in vivo pharmacokinetics of trastuzumab to yield high-contrast immuno-PET images.

The prolonged blood circulation of the radiolabeled antibody conjugates is problematic when using immuno-PET imaging due to the increased radiation exposure and longer hospitalization required until sufficient contrast develops. In contrast to the prevailing belief that PEGylation prolongs blood retention time, we observed that a PEGylated antibody with a short PEG8 linker cleared much faster from the blood while maintaining tumor uptake compared to its non-PEGylated counterpart. Breast tumors were clearly visualized with a very high tumor-to-background ratio as early as 24 h after injection in immuno-positron emission tomography (PET) imaging.

Correction: A short PEG linker alters the in vivo pharmacokinetics of trastuzumab to yield high-contrast immuno-PET images.

Correction for 'A short PEG linker alters the in vivo pharmacokinetics of trastuzumab to yield high-contrast immuno-PET images' by Woonghee Lee et al., J. Mater. Chem. B, 2021, 9, 2993-2997, DOI: 10.1039/D0TB02911D.

Indirect effects of decompression surgery on glycemic homeostasis in patients with type 2 diabetes mellitus and lumbar spinal stenosis.

BACKGROUND CONTEXT: Lumbar spinal stenosis (LSS) patients with diabetes mellitus (DM) are presumed to experience difficulty when performing regular daily exercise, although such exercise is of paramount importance for glucose homeostasis and control. Therefore, decompression surgery, which can help patients perform regular physical activity, would have indirect positive effects on blood glucose control in LSS patients with DM. PURPOSE: To evaluate the indirect effects of spinal surgery on hemoglobin A(1c) (HbA(1c)) levels in the patient with Type 2 DM and LSS. STUDY DESIGN: Prospectively collected observational cohort data. PATIENT SAMPLE: Patients with degenerative LSS and DM. OUTCOME MEASURES: The fasting total cholesterol (TC), fasting blood glucose (FBG), and HbA1c levels and visual analog scale (VAS) for back pain, VAS for leg pain, and Oswestry Disability Index (ODI). METHODS: According to the treatment methods, 31 and 37 patients were allocated to the surgical and conservative treatment groups, respectively. The HbA(1c), TC, and FBG levels and the ODI and VAS for back/leg pain were recorded for all patients before surgical and conservative treatments. At the first and second follow-up assessments after surgical or conservative treatment, the data were reassessed for all patients. RESULTS: In both groups, the VAS for back/leg pain and the ODI scores significantly decreased after surgical or conservative treatment. In the surgical treatment group, the HbA(1c) levels were significantly decreased at the first and second assessments after surgery, whereas the conservative treatment group did not show significant reductions in HbA(1c) levels at the first and second follow-up assessments. In both groups, the FBG levels did not differ between the initial and follow-up assessments. The TC levels were significantly decreased at the second follow-up assessment, only in the surgical treatment group. The amount of ODI score reduction correlated positively with the degree of HbA(1c) level reduction at the first follow-up assessment. CONCLUSIONS: The present study demonstrates the reduction in HbA(1c) level in patients with DM and LSS after decompression surgery with or without fusion. We believe this reduction in the HbA(1c) level may be a result of increased physical activity, subsequent to successful surgical decompression of the cauda equina.

Mortality after hip fractures in nonagenarians.

OBJECTIVES: Nonagenarians with hip fractures represent a special group of people because of their advanced age and co-morbidities. We evaluated mortality after hip fractures in nonagenarians. METHODS: Fifty-one patients were studied over a 2-year period. There were 39 female and 12 male patients. Twenty seven patients sustained an intertrochanteric fracture of the femur, 24 suffered from femoral neck fracture. The American Society of Anaesthetists (ASA) score of II was the most frequent among 51. Forty-one of them had one or more co-morbidities. Patient review was done 2 years after the fracture. RESULTS: The mortality rate at one year was 53.4% in men, and 15.7% in women. After multivariate analysis, the type of fracture (intertrochanteric fracture) was identified as a risk factor for one-year mortality (P = 0.025). CONCLUSION: The outcome in nonagenarians with hip fractures is poor due to the high rates of mortality, especially in men, and this could be informed to patients and their families before hip fracture surgery.

Round-robin test on thermal conductivity measurement of ZnO nanofluids and comparison of experimental results with theoretical bounds.

Ethylene glycol (EG)-based zinc oxide (ZnO) nanofluids containing no surfactant have been manufactured by one-step pulsed wire evaporation (PWE) method. Round-robin tests on thermal conductivity measurements of three samples of EG-based ZnO nanofluids have been conducted by five participating labs, four using accurate measurement apparatuses developed in house and one using a commercial device. The results have been compared with several theoretical bounds on the effective thermal conductivity of heterogeneous systems. This study convincingly demonstrates that the large enhancements in the thermal conductivities of EG-based ZnO nanofluids tested are beyond the lower and upper bounds calculated using the models of the Maxwell and Nan et al. with and without the interfacial thermal resistance.