Transcranial application of magnetic pulses for improving brain drug delivery efficiency via intranasal injection of magnetic nanoparticles.

As the blood-brain barrier (BBB) hinders efficient drug delivery to the brain, drug delivery via the intranasal pathway, bypassing the BBB, has received considerable attention. However, intranasal administration still has anatomical and physiological limitations, necessitating further solutions to enhance effectiveness. In this study, we used transcranial magnetic stimulation (TMS) on fluorescent magnetic nanoparticles (MNPs) of different sizes (50, 100, and 300 nm) to facilitate MNP's transportation and delivery to the brain parenchyma. To validate this concept, anesthetized rats were intranasally injected with the MNPs, and TMS was applied to the center of the head. As the result, a two-fold increase in brain MNP delivery was achieved using TMS compared with passive intranasal administration. In addition, histological analysis that was performed to investigate the safety revealed no gross or microscopic damages to major organs caused by the nanoparticles. While future studies should establish the delivery conditions in humans, we expect an easy clinical translation in terms of device safety, similar to the use of conventional TMS. The strategy reported herein is the first critical step towards effective drug transportation to the brain.

Development of an optical sandwich ELONA using a pair of DNA aptamers for yellow fever virus NS1.

Here, we proposed an enzyme-linked oligonucleotide assay (ELONA) for yellow fever (YF) diagnosis that uses a pair of aptamers, YFns1-4 and YFns1-31. The aptamers were selected to specifically bind to nonstructural protein 1 (NS1), which is secreted at a high concentration after YF infection. We applied the aptamers which did not interfere with each other on binding to the NS1 in a sandwich ELONA. In the assay, the best detection sensitivity was obtained when the combination of YFns1-31 as a capture aptamer and YFns1-4 as a detect aptamer was used. The sensitivity could be attributed to the results of the direct ELONA with each YFns1-4 and YFns1-31; a great absorbance intensity and a broad detectable range of NS1, respectively. The sandwich ELONA achieved a low detection limit of 0.85 nM in buffer and was highly specific to the YFV-NS1 as its detection signals were significantly distinct from those of other flavivirus-derived NS1. In addition, the assay showed a desirable sensitivity in serum-spiked condition. Our developed sandwich ELONA can be a new practical and applicable serological diagnostics in YF endemic regions where other flaviviruses coexist and facilities for complex diagnostic tests are lacking.

Aptamer-antibody hybrid ELONA that uses hybridization chain reaction to detect a urinary biomarker EN2 for bladder and prostate cancer.

We report an EN2-specific (Kd = 8.26 nM) aptamer, and a sensitive and specific enzyme-linked oligonucleotide assay (ELONA) for rapid and sensitive colorimetric detection of bladder and prostate cancer biomarker EN2 in urine. The assay relies on an aptamer-mediated hybridization chain reaction (HCR) to generate DNA nanostructures that bind to EN2 and simultaneously amplify signals. The assay can be performed within 2.5 h, and has a limit of detection of 0.34 nM in buffer and 2.69 nM in artificial urine. Moreover, this assay showed high specificity as it did not detect other urinary proteins, including biomarkers of other cancers. The proposed ELONA is inexpensive, highly reproducible, and has great chemical stability, so it may enable development of a simple, sensitive and accurate diagnostic tool to detect bladder and prostate cancers early.

Static and Dynamic Quadriceps Stretching Exercises in Patients With Patellofemoral Pain: A Randomized Controlled Trial.

BACKGROUND: Limited data are available on the effect of stretching exercise in patients with patellofemoral pain (PFP) who have inflexible quadriceps, which is one of the various causes of PFP syndrome. This study compares quadriceps flexibility, strength, muscle activation time, and patient-reported outcomes after static and dynamic quadriceps stretching exercises in patients with PFP who had inflexible quadriceps. HYPOTHESIS: Quadriceps flexibility and strength, muscle activation time, and patient-reported outcomes would improve with dynamic quadriceps stretching as compared with static quadriceps stretching exercises. STUDY DESIGN: Randomized controlled trial. LEVEL OF EVIDENCE: Level 2. METHODS: Of the 44 patients included in the study, 20 performed static stretching and 24 performed dynamic stretching. Quadriceps flexibility was assessed by measuring the knee flexion angle during knee flexion in the prone position (the Ely test). Muscle strength and muscle activation time were measured using an isokinetic device. The patient-reported outcomes were evaluated using the visual analogue scale for pain and anterior knee pain scale. RESULTS: No significant differences in quadriceps flexibility and strength, muscle activation time, and patient-reported outcomes in the involved knees were found between the 2 groups (P values > 0.05). CONCLUSION: Quadriceps flexibility and strength, muscle activation time, and patient-reported outcomes in patients with PFP who had inflexible quadriceps showed no significant differences between the static and dynamic quadriceps stretching exercise groups. CLINICAL RELEVANCE: Both static and dynamic stretching exercises may be effective for improving pain and function in patients with PFP who have inflexible quadriceps.

Effects of Static and Dynamic Stretching With Strengthening Exercises in Patients With Patellofemoral Pain Who Have Inflexible Hamstrings: A Randomized Controlled Trial.

BACKGROUND: Patellofemoral pain (PFP) syndrome is closely associated with muscle tightness. However, studies regarding the effects of stretching exercises on PFP patients with inflexible hamstrings are scarce. The aim of the study was to compare the effects between static and dynamic hamstring stretching in patients with PFP who have inflexible hamstrings. HYPOTHESIS: Compared with static hamstring stretching, dynamic hamstring stretching will improve the parameters of hamstring flexibility, knee muscle strength, muscle activation time, and clinical outcomes in this patient population. STUDY DESIGN: Prospective randomized controlled trial. LEVEL OF EVIDENCE: Level 2. METHODS: A total of 46 patients (25, static stretching; 21, dynamic stretching) participated. Hamstring flexibility was assessed according to the popliteal angle during active knee extension. Muscle strength and muscle activation time were measured using an isokinetic device. Clinical outcomes were evaluated using the visual analog scale (VAS) for pain and the anterior knee pain scale (AKPS). RESULTS: There were no differences in hamstring flexibility and knee muscle strength of the affected knees between the groups (P > 0.05). Significantly improved muscle activation time and clinical outcomes of the affected knees were observed in the dynamic stretching group compared with the static stretching group (all Ps < 0.01 for hamstring, quadriceps, VAS, and AKPS). CONCLUSION: In patients with PFP who have inflexible hamstrings, dynamic hamstring stretching with strengthening exercises was superior for improving muscle activation time and clinical outcomes compared with static hamstring stretching with strengthening exercises. CLINICAL RELEVANCE: Clinicians and therapists could implement dynamic hamstring stretching to improve function and reduce pain in patients with PFP who have inflexible hamstrings.

Poorer dynamic postural stability in patients with anterior cruciate ligament rupture combined with lateral meniscus tear than in those with medial meniscus tear.

BACKGROUND: Only limited data are available regarding postural stability between anterior cruciate ligament (ACL)-injured patients with medial meniscus (MM) tear and those with lateral meniscus (LM) tear. The purpose of this study was to compare preoperative postural stability for both involved and uninvolved knees in ACL rupture combined with MM and LM tears. It was hypothesized that there would be a significant difference in postural stability between these two groups. METHODS: Ninety-three ACL-injured patients (53 combined with MM tears vs. 40 combined with LM tears) were included. Static and dynamic postural stability were evaluated with the overall stability index (OSI), anterior-posterior stability index (APSI), and medial-lateral stability index (MLSI) using stabilometry. Knee muscle strength was evaluated using an isokinetic testing device. RESULTS: In the static postural stability test, none of the stability indices showed significant differences between the two groups for both knees (p > 0.05). In the dynamic postural stability test for involved side knees, the OSI and APSI were significantly higher in the LM tear group compared to the MM tear group (OSI: 2.0 ± 0.8 vs. 1.6 ± 0.5, p = 0.001; APSI: 1.5 ± 0.6 vs. 1.3 ± 0.5, p = 0.023), but not the MLSI (p > 0.05). In the static and dynamic postural stability tests in each group, there were no significant differences between the involved and uninvolved side knees (p > 0.05). There was no significant difference in the knee muscle strength between the two groups (p > 0.05). All postural stability showed no significant correlation with knee muscle strength (p > 0.05). CONCLUSION: Dynamic postural stability was poorer in patients with ACL rupture combined with LM tear than in those with MM tear. Therefore, close monitoring for postural stability would be necessary during preoperative and postoperative rehabilitation, especially for patients with ACL rupture combined with LM tear. LEVEL OF EVIDENCE: LEVEL III.

Abiotic and biotic stress tolerance in Arabidopsis overexpressing the multiprotein bridging factor 1a (MBF1a) transcriptional coactivator gene.

We conducted a genetic yeast screen to identify salt tolerance (SAT) genes in a maize kernel cDNA library. During the screening, we identified a maize clone (SAT41) that seemed to confer elevated salt tolerance in comparison to control cells. SAT41 cDNA encodes a 16-kDa protein which is 82.4% identical to the Arabidopsis Multiprotein bridging factor 1a (MBF1a) transcriptional coactivator gene. To further examine salinity tolerance in Arabidopsis, we functionally characterized the MBF1a gene and found that dehydration as well as heightened glucose (Glc) induced MBF1a expression. Constitutive expression of MBF1a in Arabidopsis led to elevated salt tolerance in transgenic lines. Interestingly, plants overexpressing MBF1a exhibited insensitivity to Glc and resistance to fungal disease. Our results suggest that MBF1a is involved in stress tolerance as well as in ethylene and Glc signaling in Arabidopsis.

Dual positional specificity and expression of non-traditional lipoxygenase induced by wounding and methyl jasmonate in maize seedlings.

Lipoxygenases (LOXs) catalyze the formation of fatty acid hydroperoxides involved in responses to stresses. This study examines the expression of a non-traditional dual positional specific maize LOX in response to wounding or methyl jasmonate (MeJA). Full-length maize LOX cDNA was expressed in Escherichia coli, and recombinant LOX was purified and characterized enzymatically. RP-HPLC and GC-MS analysis showed that the purified LOX converts alpha-linolenic acid into 13-hydroperoxylinolenic acid and 9-hydroperoxylinolenic acid in a 6:4 ratio. LOX mRNA accumulated rapidly and transiently in response to wounding reaching a peak of expression about 3 h after wounding. This increase followed an initial increase in endogenous jasmonic acid (JA) 1 h after wounding (JA burst). However, the expression of LOX induced by MeJA lasted longer than the expression induced by wounding, and the MeJA-induced expression seemed to be biphasic pattern composed of early and late phases. The expression of LOX in the presence of inhibitors of JA biosynthesis was not completely inhibited, but delayed in wound response and the expression period was shortened in MeJA response. These results suggest that wound-responsive JA burst may trigger the early phase of LOX expression which facilitates biosynthesis of endogenous JA through its 13-LOX activity, and subsequently leads to the activation of the late phase LOX expression in MeJA-treated maize seedlings. Implications of dual positional specificity of maize LOX in the observed expression kinetics are discussed.

A point mutation of valine-311 to methionine in Bacillus subtilis protoporphyrinogen oxidase does not greatly increase resistance to the diphenyl ether herbicide oxyfluorfen.

In an effort to asses the effect of Val311Met point mutation of Bacillus subtilis protoporphyrinogen oxidase on the resistance to diphenyl ether herbicides, a Val311Met point mutant of B. subtilis protoporphyrinogen oxidase was prepared, heterologously expressed in Escherichia coli, and the purified recombinant Val311Met mutant protoporphyrinogen oxidase was kinetically characterized. The mutant protoporphyrinogen oxidase showed very similar kinetic patterns to wild type protoporphyrinogen oxidase, with slightly decreased activity dependent on pH and the concentrations of NaCl, Tween 20, and imidazole. When oxyfluorfen was used as a competitive inhibitor, the Val311Met mutant protoporphyrinogen oxidase showed an increased inhibition constant about 1.5 times that of wild type protoporphyrinogen oxidase. The marginal increase of the inhibition constant indicates that the Val311Met point mutation in B. subtilis protoporphyrinogen oxidase may not be an important determinant in the mechanism that protects protoporphyrinogen oxidase against diphenyl ether herbicides.