Pretreatment of rhesus monkeys with transdermal patches containing physostigmine and procyclidine: implications of the delivery system for the potential application against VX nerve agent intoxication in humans.

Physostigmine (Phs) is a reversible inhibitor of acetylcholinesterase (AChE) that penetrates the blood-brain barrier (BBB) and could be used to protect the central nervous system (CNS) against the effects of nerve agents. For prophylactic effectiveness, long, steady, and adequate inhibition of AChE activity by Phs is needed to broadly protect against the CNS effects of nerve agents. Here, we evaluated the efficacy of transdermal patches containing Phs and procyclidine (PC) as prophylactic agents. Patches (25 cm2) containing 4.4 mg Phs and 17.8 mg PC had a protective ratio of approximately 78.6-fold in rhesus monkeys challenged with VX nerve agent and given an antidote. Physiologically based pharmacokinetic model in conjunction with an indirect pharmacodynamic (PBPK/PD) was developed for Phs and scaled to rhesus monkeys. The model was able to reproduce the concentration profile and inhibitory effect on AChE of Phs in monkeys, as evidenced by correlation coefficients of 0.994 and 0.992 for 25 cm2 and 49 cm2 patches, respectively (i.e., kinetic data), and 0.989 and 0.968 for 25 cm2 and 49 cm2 patches, respectively (i.e., dynamic data). By extending the monkey PBPK/ PD model to humans, the effective human dose was predicted to be five applications of a 25 cm2 patch (i.e., 22 mg Phs), and two applications of a 49 cm2 patch (i.e., 17.4 mg Phs). Therefore, given that patch application of Phs in rhesus monkeys has a prolonged effect (namely, AChE inhibition of 19.6% for the 25 cm2 patch and 23.0% for the 49 cm2 patch) for up to 216 h, patch formulation of Phs may provide similar protection against nerve agent intoxication in humans.

Differences in the 3-dimensional aging changes of the lips among female adults with skeletal Class I, II, and III malocclusion.

INTRODUCTION: The objective of this study was to evaluate and compare the 3-dimensional (3D) aging changes of the lips among adult skeletal Class I, II, and III malocclusion. METHODS: Female adult orthodontic patients aged 20-50 years with pretreatment cone-beam computed tomography scans were retrospectively classified according to age (20s [20-29 years], 30s [30-39 years], and 40s [40-49 years]) and then subclassified by malocclusion into skeletal Class I, II, and III relationship (9 groups; n = 30 per group). Positional differences in midsagittal and parasagittal soft-tissue landmarks and 3D morphologic aging changes of the lips were evaluated using cone-beam computed tomography scans. RESULTS: Labiale superius and cheilion for patients in their 40s indicated a significant downward and backward position compared with those in their 20s, regardless of skeletal classifications (P <0.05). Accordingly, the upper lip height decreased, and the mouth width increased significantly (P <0.05). For Class III malocclusion, the upper lip vermilion angle was greater for patients in their 40s than those in their 20s (P <0.05), whereas the lower lip vermilion angle was only lower for patients with Class II malocclusion (P <0.05). CONCLUSIONS: Middle-aged adult females (40-49 years) had a lower upper lip height and greater mouth width than those in their 20s, regardless of skeletal malocclusion. However, prominent morphologic aging changes of the lips were noted on the upper lip for skeletal Class III malocclusion and the lower lip for skeletal Class II malocclusion, implying that the underlying skeletal features (or malocclusion) may influence 3D aging changes of the lips.

Comparison of Lip Line Cant Change After 1-Jaw and 2-Jaw Surgery.

OBJECTIVES: The purpose of this study was to compare the lip line cant (LLC) changes after 1 and 2-jaw surgery, and to evaluate the correlations of the craniofacial factors affecting LLC. METHODS: The study subjects were selected (LLC amount within 1.5-6.0°) from among the patients diagnosed with Class III malocclusion who underwent one (n = 20) or 2-jaw surgery (n = 20). Cone beam computed tomography images were obtained immediately before the operation (T1) and 6 months after the operation (T2). Preoperative and postoperative craniofacial measurements were obtained. RESULTS: The study subjects showed 3.12° LLC on average before undergoing 1-jaw surgery, and their LLC changed to 1.27° after the surgery. As for 2-jaw surgery, the subjects showed 3.38° LLC on average before the surgery and 0.98° after the surgery. LLC at pre-treatment may be more affected by a cant of the occlusal plane in the mandible than maxilla. In the comparison of the value of changes of LLC, the value of 2-jaw surgery was bigger than the value of 1-jaw surgery but the difference was statistically insignificant. LIMITATIONS: This study had a limitation in that the muscles were not considered. And the metal bracket or metal crown and bridge, however, can cause noise and blurring artifacts in CT, which can lead to a low resolution. And the limited number of the patients should be considered. CONCLUSIONS: In correlation analysis, both pre-surgery LLC and change of LLC have correlation with almost all the craniofacial measurement. Lip-line cant of patients with facial asymmetry has higher correlation with mandibular cant than with other cants. To improve the LLC, a surgical plan should be established to minimize the mandibular cant using the computer simulation as well as the maxillary cant.

Salivary glutamate is elevated in individuals with chronic migraine.

Background Glutamate has been implicated in migraine pathogenesis, and is elevated in the plasma, cerebrospinal fluid, and saliva in migraineurs. However, no comparison of glutamate levels among chronic migraine, episodic migraine and controls has been reported. The aim is to compare salivary glutamate levels of individuals with chronic migraine with those of individuals with episodic migraine and healthy controls. Methods We investigated salivary glutamate level of 46 women with chronic migraine, 50 women with episodic migraine, and 19 healthy controls via enzyme linked immunosorbent assay. Results The salivary glutamate level of the chronic migraine group (median and interquartile range, 20.47 [15.27-30.15] pmol/mg total protein) was significantly higher than those of the episodic migraine (16.17 [12.81-20.15] pmol/mg total protein, p = 0.008) and control (12.18 [9.40-16.24] pmol/mg total protein, p = 0.001) groups. The salivary glutamate level of the episodic migraine group was marginally elevated from that of the control group (post hoc p = 0.016). Thresholds of 16.58 and 17.94 pmol/mg total protein optimize the sensitivity and specificity to differentiate chronic migraine participants from healthy controls and episodic migraine participants, respectively. Conclusions Salivary glutamate level was elevated in chronic migraine participants. These data suggest that salivary glutamate level could be an indicator of CM.

3,4-Dihydroquinazoline derivatives inhibit the activities of cholinesterase enzymes.

A series of 3,4-dihydroquinazoline derivatives consisting of the selected compounds from our chemical library on the diversity basis and the new synthetic compounds were in vitro tested for their inhibitory activities for both acetylcholinesterase (AChE, from electric eel) and butyrylcholinesterase (BChE, from equine serum) enzymes. It was discovered that most of the compounds displayed weak AChE and strong BuChE inhibitory activities. In particular, compound 8b and 8d were the most active compounds in the series against BChE with IC50 values of 45nM and 62nM, as well as 146- and 161-fold higher affinity to BChE, respectively. To understand the excellent activity of these compounds, molecular docking simulations were performed to get better insights into the mechanism of binding of 3,4-dihydroquinazoline derivatives. As expected, compound 8b and 8d bind to both catalytic anionic site (CAS) and peripheral site (PS) of BChE with better interaction energy values than AChE, in agreement with our experimental data. Furthermore, the non-competitive/mixed-type inhibitions of both compounds further confirmed their dual binding nature in kinetic studies.

Effect of the T-type channel blocker KYS-05090S in mouse models of acute and neuropathic pain.

T-type channels are important contributors to the initiation and the maintenance of chronic pain states. Blocking T-type channels is therefore a possible therapeutic strategy for relieving pain. Here, we report the Cav3.2 T-type channel blocking action of a previously reported small organic molecule, KYS-05090S. This compound was able to reduce transiently expressed Cav3.2 currents with low micromolar affinity and mediated a hyperpolarizing shift in half-inactivation potential. KYS-05090S was then tested in models of acute and neuropathic pain. KYS-05090S (10 µg/10 µl delivered intrathecally) significantly reduced acute pain induced by formalin in both the tonic and inflammatory phases. Its antinociceptive effect was not observed when delivered to Cav3.2 null-mice revealing a Cav3.2-dependent mechanism. KYS-05090S also reduced neuropathic pain in a model of partial sciatic nerve injury. Those results indicate that KYS-05090S mediates a potent analgesic effect in inflammatory and neuropathic pain through T-type channel modulation, suggesting that its scaffold could be explored as a new class of analgesic compounds.

In vitro synergistic anticancer activity of the combination of T-type calcium channel blocker and chemotherapeutic agent in A549 cells.

As a result of our continuous research, new 3,4-dihydroquinazoline derivative containing ureido group, KCP10043F was synthesized and evaluated for T-type Ca(2+) channel (Cav3.1) blockade, cytotoxicity, and cell cycle arrest against human non-small cell lung (A549) cells. KCP10043F showed both weaker T-type Ca(2+) channel blocking activity and less cytotoxicity against A549 cells than parent compound KYS05090S [4-(benzylcarbamoylmethyl)-3-(4-biphenylyl)-2-(N,N',N'-trimethyl-1,5-pentanediamino)-3,4-dihydroquinazoline 2 hydrochloride], but it exhibited more potent G1-phase arrest than KYS05090S in A549 cells. This was found to be accompanied by the downregulations of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D2, cyclin D3, and cyclin E at the protein levels. However, p27(KIP1) as a CDK inhibitor was gradually upregulated at the protein levels and increased recruitment to CDK2, CDK4 and CDK6 after KCP10043F treatment. Based on the strong G1-phase cell cycle arrest of KCP10043F in A549 cells, the combination of KCP10043F with etoposide (or cisplatin) resulted in a synergistic cell death (combination index=0.2-0.8) via the induction of apoptosis compared with either agent alone. Taken together with these overall results and the favorable in vitro ADME (absorption, distribution, metabolism, and excretion) profiles of KCP10043F, therefore, it could be used as a potential agent for the combination therapy on human lung cancer.

Direct-write patterning of bacterial cells by dip-pen nanolithography.

The ability of dip-pen nanolithography (DPN) to generate nano- or microarrays of soft or hard materials (e.g., small molecules, DNA, proteins, nanoparticles, sols, and polymers) in a direct-write manner has been widely demonstrated. The transporting of large-sized ink materials such as bacteria, however, remains a significant challenge with this technique. The size limitation of the water meniscus formed between the DPN tip and the solid surface becomes a bottleneck in such diffusion-based molecular transport experiments. Herein, we report a straightforward "stamp-on" DPN method that uses a nanostructured poly(2-methyl-2-oxazoline) hydrogel-coated tip and carrier agents to generate patterns of micrometer-sized Escherichia coli JM 109 bacterial cells. We demonstrate that this approach enables the deposition of a single bacterial cell array on a solid surface or arrays of layers of multiple cells by modulating the viscosity of the "ink" solution. Fluorescence microscopy images indicated that the deposited bacterial cells were kept alive on Luria-Bertani-agar layered solid surfaces after DPN patterning.

Therapeutic Efficacy of ABN401, a Highly Potent and Selective MET Inhibitor, Based on Diagnostic Biomarker Test in MET-Addicted Cancer.

The receptor tyrosine kinase c-MET regulates processes essential for tissue remodeling and mammalian development. The dysregulation of c-MET signaling plays a role in tumorigenesis. The aberrant activation of c-MET, such as that caused by gene amplification or mutations, is associated with many cancers. c-MET is therefore an attractive therapeutic target, and inhibitors are being tested in clinical trials. However, inappropriate patient selection criteria, such as low amplification or expression level cut-off values, have led to the failure of clinical trials. To include patients who respond to MET inhibitors, the selection criteria must include MET oncogenic addiction. Here, the efficacy of ABN401, a MET inhibitor, was investigated using histopathologic and genetic analyses in MET-addicted cancer cell lines and xenograft models. ABN401 was highly selective for 571 kinases, and it inhibited c-MET activity and its downstream signaling pathway. We performed pharmacokinetic profiling of ABN401 and defined the dose and treatment duration of ABN401 required to inhibit c-MET phosphorylation in xenograft models. The results show that the efficacy of ABN401 is associated with MET status and they highlight the importance of determining the cut-off values. The results suggest that clinical trials need to establish the characteristics of each sample and their correlations with the efficacy of MET inhibitors.

How assistive devices affect activities of daily living and cognitive functions of people with brain injury: a meta-analysis.

PURPOSE: This study aims to systematically evaluate and synthesize the literature on the effects of assistive technology devices on the activities of daily living and cognitive functions of people with brain injury. METHODS: Eight randomized controlled trials were selected from online databases pertaining to the scientific use of AT devices for people with brain injury. The Jadad scale was used to analyse the subjects qualitatively, and Comprehensive Meta-Analysis 2.0 was used to test the statistical heterogeneity, effect size, sensitivity and publication bias of each of the selected studies. RESULTS: All selected studies were assigned a score of three on the Jadad scale, which could classify them as high-quality studies. The total number of participants in the studies was 385. The effect size of activities of daily living was 0.72, which is a medium effect size and that of cognitive function was 0.30, which is a small effect size. CONCLUSION: Assistive devices are effective in improving the activities of daily living and relatively less effective in enhancing the cognitive function of people with brain injury. This meta-analysis is evidence that assistive devices could be an effective intervention method for people with brain injury. Implications for Rehabilitation The purpose of this study is to demonstrate the effectiveness of this approach and to generalize the use of assistive devices. We aim to provide a basis for popularizing assistive devices as a therapeutic intervention method.