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Ionic liquids (ILs) are known to provide stability to biomolecules. ILs are also widely used in the fields of chemical engineering, biological engineering, chemistry, and biochemistry because they facilitate enzyme catalyzed reactions and enhance their conversion rate. In this work, we have evaluated the influence of alkyl chain substitution of ammonium ILs such as diethylammonium dihydrogen phosphate (DEAP) and triethylammonium hydrogen phosphate (TEAP) for the stability and activity of the tobacco etch virus (TEV) protease. Further, we performed molecular dynamics (MD) simulations to calculate the RMSD (root mean square deviation) for TEV and TEV + ILs. Experimental and simulations results show that TEV is more stable in the presence of TEAP than DEAP. Whereas, TEV protease activity for the cleavage of fusion proteins is preserved in the presence of DEAP while lost in the presence of TEAP. Hence, DEAP IL can serve as alternative solvents for the stability of the TEV protease with preserved activity. To the best of our knowledge, this is first study to show that ILs can stabilize and maintain the TEV protease cleavage activity.

The seat comfort of automobiles is one of the significant factors for determining the driver’s fatigue, emotional experience, and individual space (which captures their individuality, rather than just a means of transportation in modern society). Conventional automobile seats could not provide seating comfort suitable for all drivers, in the form of seats that fit each driver’s body type and the difficulty of meeting individual needs. This study proposes self-shape adjustable (the SSA seats) seats that improve the sitting comfort, safety, and secure the stability, by adjusting shape fit to the driver’s body type. The SSA seats transforms the seat itself, in a way that improves the distribution of contact pressure and reduces sitting fatigue, with the pneumatic system. The transformed seats provide better sitting comfort and safety than the conventional automobile seat, by providing a seat shape suitable for the body shape of all users. It was verified that the SSA seats, proposed in this paper, have a uniform and improved pressure distribution, compared to the conventional seat, in various sitting postures; the contact area between the seat and user is enlarged, and the pressure concentrated on the ischial bone is lowered. In addition, it was proven (through user evaluation) that quantitative evaluation verification was the same as qualitative evaluation results.

Oxidative stress is one of the leading causes of various diseases. Oxidative stress conditions occur when reactive oxygen species (ROS) levels exceed cell defense mechanisms. Antioxidants are effective against ROS, a major factor in oxidative stress. Antioxidants play important roles in our body by scavenging free radicals, thereby preventing them from damaging biological molecules. Vitamin C is essential for various physiological functions such as the synthesis of collagen and neurotransmitters, and it is a major antioxidant. Although vitamin C is widely used in cosmetic and therapeutic products, it easily undergoes oxidation by air, pH, temperature, and UV light, thereby decreasing its potency as an antioxidant and reducing the shelf-life of products containing vitamin C. To overcome this drawback, we developed Aptamin C®, an innovative single-strand DNA aptamer that maximizes the antioxidant efficacy of vitamin C by binding to its reduced form and delaying its oxidation. To investigate the effect of Aptamin C® and vitamin C complex on human skin, we performed both in vitro and clinical tests. We observed that the Aptamin C® and vitamin C complex had effective ROS-scavenging and anti-inflammatory effects.

Ionic polymer metal composites with a flexible large deformation have been used as biomimetic actuators and sensors in various fields. This work mainly focuses on the validation of the proposed the...

Dishevelled (Dvl) plays a crucial role in Wnt signaling by interacting with membrane-bound receptors and downstream molecules through its PDZ domain. CXXC5 is one of the key molecules that interacts with Dvl and negatively regulates the Wnt/β-catenin pathway in osteoblast differentiation. Recently, the Dvl-CXXC5 interaction has been identified as an excellent target for osteoporosis treatment. Therefore, it is desirable to have detailed structural information for the Dvl-CXXC5 interaction. Although solution structures of the Dvl1 PDZ domain have been reported, a high-resolution crystal structure would provide detailed sidechain information that is essential for drug development. Here, we determined the first crystal structure of the Dvl-1 PDZ domain at a resolution of 1.76 Å, and compared it with its previously reported solution structure. The Dvl1 PDZ domain crystal belonged to the space group H32 with unit-cell parameters a = b = 72.837, c = 120.616, α = β = 90.00, γ = 120.00. The crystal structure of Dvl1 PDZ shared its topology with the previously reported structure determined by nuclear magnetic resonance (NMR); however, the crystal structure was quite different from the solution structure in both the secondary structural region and the ligand-binding pocket. Molecular modeling based on NMR and X-ray crystallographic data yielded detailed information about the Dvl1/CXXC5 interaction, which will be useful for designing inhibitors.

RAS proteins play critical roles in various cellular processes, including growth and transformation. RAS proteins are subjected to protein stability regulation via the Wnt/β-catenin pathway, and glycogen synthase kinase 3 beta (GSK3β) is a key player for the phosphorylation-dependent RAS degradation through proteasomes. GSK3β-mediated RAS degradation does not occur in cells that express a nondegradable mutant (MT) β-catenin. Here, we show that β-catenin directly interacts with RAS at the α-interface region that contains the GSK3β phosphorylation sites, threonine 144 and threonine 148 residues. Exposure of these sites by prior β-catenin degradation is required for RAS degradation. The introduction of a peptide that blocks the β-catenin-RAS interaction by binding to β-catenin rescues the GSK3β-mediated RAS degradation in colorectal cancer (CRC) cells that express MT β-catenin. The coregulation of β-catenin and RAS stabilities by the modulation of their interaction provides a mechanism for Wnt/β-catenin and RAS-ERK pathway cross-talk and the synergistic transformation of CRC by both APC and KRAS mutations.

The variation in the biological function of proteins plays an important role in plasma medicine and sterilization. Several non-thermal plasma sources with different feeding gases are used worldwide for plasma treatment, including dielectric barrier discharge (DBD) and atmospheric-pressure plasma jet (APPJ) as the most commonly used sources. Therefore, in the present work, we used both DBD and APPJ plasma sources with N2 and air as feeding gases to evaluate the effects on the structural, thermodynamic, and activity changes of enzymes. In the current work, we used lysozyme as a model enzyme and verified the structural changes using circular dichroism (CD), fluorescence, and X-ray crystallography. In addition, we investigated the lysozyme thermodynamics using CD thermal analysis and changes in the B-factor from X-ray crystallography. The results showed that lysozyme activity decreased after the plasma treatment. From these analyses, we concluded that N2-feeding gas plasma disturbs the structure and activity of lysozyme more than Air feeding gas plasma in our experimental studies. This study provides novel fundamental information on the changes to enzymes upon plasma treatment, which has been absent from the literature until now.

Surgical exposure for massive tumor of the anterior skull base can be difficult. We have combined two surgical approaches, the subfrontal/subcranial approach and the Le Fort I osteotomy approach, to provide panoramic accesses to the anterior skull base. Via this combination of exposures, we have easily accessed and resected tumors extending from the cribiform plate all the way down to the level of the upper cervical spine. Additionally, we have used the nasal endoscope and microscope to assist in our resection. This approach has several advantages, which include ease of post-resection skull base reconstruction and excellent cosmetic results by avoiding anterior facial incisions. We believe this approach should be in the armamentarium of any surgeon resecting massive anterior skull base tumor.

To study the efficacy of vagus nerve stimulation (VNS) therapy as an adjunctive treatment for intractable epilepsy in patients under 12 years of age, we analyzed 2-year postimplant data of 35 consecutive patients. Of the 35 patients, 18 (51.4%) at 6 months, 18 (51.4%) at 12 months, and 21 (60.1%) at 24 months showed ≥50% reduction in seizure frequency (responders). Although incremental seizure freedom was noted, no patient remained seizure-free throughout the 3 study periods. Partial response (≥50% seizure reduction in 2 or less study periods) was seen in 8 (22.9%) patients. Twelve patients (34.3%) were nonresponders. Out of 29 patients with primary generalized epilepsy, 20 (68.9%) and, out of 6 patients with focal epilepsy, 3 (50%) had ≥50% seizure control in at least one study period. No major complications or side effects requiring discontinuation of VNS therapy were encountered. We conclude that (1) patients with intractable primary generalized epilepsy respond better to VNS therapy, (2) cumulative effect of neuromodulation with improving responder rate to seizure freedom with continuation of VNS therapy is noted, and (3) VNS therapy is safe and is well tolerated in children receiving implant under 12 years of age.

Both the Wnt/β-catenin and Ras pathways are aberrantly activated in most human colorectal cancers (CRCs) and interact cooperatively in tumor promotion. Inhibition of these signaling may therefore be an ideal strategy for treating CRC. We identified KY1220, a compound that destabilizes both β-catenin and Ras, via targeting the Wnt/β-catenin pathway, and synthesized its derivative KYA1797K. KYA1797K bound directly to the regulators of G-protein signaling domain of axin, initiating β-catenin and Ras degradation through enhancement of the β-catenin destruction complex activating GSK3β. KYA1797K effectively suppressed the growth of CRCs harboring APC and KRAS mutations, as shown by various in vitro studies and by in vivo studies using xenograft and transgenic mouse models of tumors induced by APC and KRAS mutations. Destabilization of both β-catenin and Ras via targeting axin is a potential therapeutic strategy for treatment of CRC and other type cancers activated Wnt/β-catenin and Ras pathways.

Plasma medicine is an upcoming research area that has attracted the scientists to explore more deeply the utility of plasma. So, apart from the treating biomaterials and tissues with plasma, we have studied the effect of soft plasma with different feeding gases such as Air, N2 and Ar on modification of biomolecules. Hence, in this work we have used the soft plasma on biomolecules such as proteins ((Hemoglobin (Hb) and Myoglobin (Mb)), calf thymus DNA and amino acids. The structural changes or structural modification of proteins and DNA have been studied using circular dichroism (CD), fluorescence spectroscopy, protein oxidation test, gel electrophoresis, UV-vis spectroscopy, dynamic light scattering (DLS) and 1D NMR, while Liquid Chromatograph/Capillary Electrophoresis-Mass Spectrometer (LC/CE-MS) based on qualitative and quantitative bio-analysis have been used to study the modification of amino acids. Further, the thermal analysis of the protein has been studied with differential scanning calorimetry (DSC) and CD. Additionally, we have performed docking studies of H2O2 with Hb and Mb, which reveals that H2O2 molecules preferably attack the amino acids near heme group. We have also shown that N2 gas plasma has strong deformation action on biomolecules and compared to other gases plasma.

Object. Transsphenoidal adenomectomy with resection of a defined pituitary adenoma has been the treatment of choice for CD for the last 30 years. Surgical resection, however, may not always result in long-term remission of CD. This is particularly important in light of the high risk of morbidity and mortality in patients in the unsuccessfully treated cushingoid state. As such, it is interesting to identify prognostic factors that may predict the likelihood of long-term remission. Methods. The authors review their series of 174 patients who have undergone transsphenoidal procedures for CD over a period of 20 years with minimum follow-up periods of 5 years. Selection of these patients was based on clinical, imaging, and laboratory criteria that included serum cortisol levels, loss of diurnal variation in serum cortisol levels, urinary free cortisol concentration, and results of a dexamethasone suppression test, petrosal sinus sampling, and corticotroph-releasing hormone stimulation tests as indicated. All patients who met the biochemical criteria underwent transsphenoidal microsurgery. The authors found an overall rate of remission of 74% at 5 years postoperatively. Patients in whom morning serum cortisol concentrations were lower than 3 µg/dl (83 nmol/L) on postoperative Day 3, following an overnight dexamethasone suppression test, had a 93% chance of remission at the 5-year follow-up examination. Patients with cortisol concentrations higher than this level uniformly failed to achieve long-term remission. Conclusions. Transsphenoidal microsurgery is an effective means of control for patients with adrenocorticotrophic hormone—producing microadenomas. Clinical outcome correlated well with the size of the tumor, as measured on preoperative imaging studies, and with postoperative morning cortisol levels following an overnight dexamethasone suppression test. Postoperative cortisol levels can be used as a useful prognostic indicator of the likelihood of future recurrence following transsphenoidal adenomectomy in CD.

To establish efficacy of vagus nerve stimulation (VNS) therapy in the treatment of intractable epilepsy, we compared outcome data from the baseline period to the maintenance period (7 to 24 months post-implant) by calculating the mean seizure frequency at 6-month intervals (7 to 12 months - first study period, 13 to 18 months - second study period, and 19 to 24 months - third study period), in 39 consecutive patients on unchanged antiepilepsy drug (AED) regimen for 24 months following the VNS implant. Of the 39 patients24 (61.5%) at first study period, 25 (64.1 %) at second study period and 25 (64.1 %) at third study period were responders (≥50% reduction in seizures). Twenty one (53.9 %) patients were responders in all three study periods. Incremental seizure control was seen in 15 of these 21 patients. Although 3 (7.7%), 4 (10.3 %) and 8 (20.5%) patients had a total (100%) seizure control at first, second and third study periods respectively, no patient remained seizure-free through all 3 study periods. Seven (17.9%) patients were partial responders (≥ 50% seizure reduction in two or less study periods). Eleven patients (28.2%) were non-responders (< 50% seizure reduction in all 3 study periods). Twenty three patients (59%) had partial epilepsy with and without secondarily generalization and 16 patients (41%) had primary generalized epilepsy. Eleven (47.8%) of the 23 patients with partial epilepsy and 10 (62.5%) of the 16 patients with generalized epilepsy were responders in all three study periods. We conclude that: 1) More than 60 % of patients on unchanged AED regimen continued to be responders at 24 months following the VNS implant. 2) A trend towards increasing responder rate with increasing duration of VNS therapy was observed.3) No major complications or side effects requiring discontinuation of VNS therapy were noted.

Interleukin-8 (IL-8) is a proinflammatory CXC chemokine that has been associated with the promotion of neutrophil chemotaxis, degranulation, and the pathogenesis of several neutrophil-infiltrating chronic inflammatory diseases. In the current study, we generated and characterized a 2'-fluoro-pyrimidine modified RNA aptamer (8A-35) against human IL-8. The 8A-35 aptamer binds to IL-8 with high specificity and affinity, yielding an estimated K(D) of 1.72 pM. NMR data revealed that the residues of Lys8, Leu10, Val63, Val66, Lys69 and Ala74 of IL-8 interact with aptamer. Moreover, the 8A-35 aptamer has a potent IL-8-neutralizing activity that can modulate multiple biological activities of IL-8 in human neutrophils, including migration, intracellular signaling, and intracellular Ca(2+) mobilization. Our results suggest that the 8A-35 aptamer has great potential to be a lead structure in the development of effective therapeutic agents against inflammatory diseases.

Objective Elemental iodine (I(2)) can kill a broad spectrum of organisms, including bacteria, fungi, and viruses. Furthermore, it is inexpensive, bacterial resistance is unknown, and allergic reactions are rare. Because of these properties, we wanted to determine the concentration of I(2) that would kill Staphylococcus epidermidis and Staphylococcus aureus without causing injury to the central nervous system, in an attempt to further reduce the rates of shunt infections. Methods Bacterial kill studies using S. epidermidis and S. aureus were performed by using Ringer's lactate solution alone or solution containing I(2) at a concentration of 5, 10, 20, 50, 100, or 1000 parts per million (ppm), cefazolin at 1 mg/ml, or bacitracin at 100 units/ml. Twenty-one adult male Wistar rats, in seven groups, underwent a frontoparietal craniectomy. The surfaces of their brains were irrigated for 1 hour with Ringer's lactate solution alone or solution containing I(2) (at the concentrations noted above). After 72 hours of observation, the animals were killed. Their brains were then fixed in formalin, stained with hematoxylin/eosin, and examined. Results Even with exposure of only 15 seconds to an I(2) solution of 20 ppm, no growth was detected with an inoculum of either bacteria of 100 million. In contrast, the two antibiotics were not nearly as effective as I(2), with kill rates ranging from 19 to 93%. Examination of the rat brains demonstrated no histological changes after subarachnoid exposure to solutions containing 5, 10, 20, or 50 ppm; however, necrosis was observed with concentrations of 100 and 1000 ppm. Conclusion I(2) can be added to irrigation solutions in sufficient concentrations to be bactericidal without causing any central nervous system injury.