Πέμπτη 28 Ιανουαρίου 2016

PAI-1, a target gene of miR-143, regulates invasion and metastasis by upregulating MMP-13 expression of human osteosarcoma

Abstract

Despite recent improvements in the therapy for osteosarcoma, 30–40% of osteosarcoma patients die of this disease, mainly due to its lung metastasis. We have previously reported that intravenous injection of miR-143 significantly suppresses lung metastasis of human osteosarcoma cells (143B) in a mouse model. In this study, we examined the biological role and mechanism of miR-143 in the metastasis of human osteosarcoma cells. We identified plasminogen activator inhibitor-1 (PAI-1) as a direct target gene of miR-143. To determine the role of PAI-1 in human osteosarcoma cells, siRNA was transfected into 143B cells for knockdown of PAI-1 expression. An in vitro study showed that downregulation of PAI-1 suppressed cell invasion activity, but not proliferation. Moreover, injection of PAI-1 siRNA into a primary lesion in the osteosarcoma mouse model inhibited lung metastasis compared to control siRNA-injected mice, without influencing the proliferative activity of the tumor cells. Subsequent examination using 143B cells revealed that knockdown of PAI-1 expression resulted in downregulation of the expression and secretion of matrix metalloproteinase-13 (MMP-13), which is also a target gene of miR-143 and a proteolytic enzyme that regulates tumor-induced osteolysis. Immunohistochemical analysis using clinical samples showed that higher miR-143 expressing cases showed poor expression of PAI-1 in the primary tumor cells. All such cases belonged to the lung metastasis-negative group. Moreover, the frequency of lung metastasis-positive cases was significantly higher in PAI-1 and MMP-13 double-positive cases than in PAI-1 or MMP-13 single-positive or double-negative cases (< 0.05). These results indicated that PAI-1, a target gene of miR-143, regulates invasion and lung metastasis via enhancement of MMP-13 expression and secretion in human osteosarcoma cells, suggesting that these molecules could be potential therapeutic target genes for preventing lung metastasis in osteosarcoma patients.

Thumbnail image of graphical abstract

Experiments conducted in vitro and in vivo and also clinical samples show that plasminogen activator inhibitor-1 (PAI-1) is related to invasion via up regulation of matrix metalloprotease 13 resulting in metastasis. The findings suggest that PAI-1 is likely to therapeutic target for preventing lung metastasis in human osteosarcoma.



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IJERPH, Vol. 13, Pages 177: Factors Associated with Relapse among Heroin Addicts: Evidence from a Two-Year Community-Based Follow-Up Study in China

Background: Many countries including China are facing a serious opiate dependence problem. Anti-drug work effectiveness was affected by the high relapse rate all over the world. This study aims to analyze the factors influencing heroin addict relapse, and to provide evidence for generating relapse prevention strategies. Methods: A community-based follow-up study was conducted in China between October 2010 and September 2012. A total of 554 heroin addicts in accordance with the inclusion criteria from 81 streets in 12 districts of Shanghai, China were divided into 4 groups: group 1—daily dosage taken orally of 60 mL of methadone or under combined with psychological counseling and social supports (n = 130); group 2—daily dosage taken orally of over 60 mL of methadone combined with psychological counseling and social supports (n = 50); group 3—JTT (Jitai tablets) combined with psychological counseling and social supports (n = 206); group 4—JTT combined with social supports (n = 168). Results: Log-rank test results showed that the cumulative relapse rate differences among four groups during the two-year follow-up period were not statistically significant (χ2 = 5.889, p = 0.117). Multivariate Cox regression analysis results showed that only three independent variables were still statistically significant, including compliance with participation in psychological counseling (OR = 3.563, p = 0.000), the years of drug use (OR = 1.078, p = 0.001)and intervention model. Conclusions: Using the detoxification medications combined with appropriate psychological counseling and social support measures will help improve the effectiveness of relapse prevention, which is a kind of alternative community detoxification pattern. Appropriate and standard psychological counseling is very important for anti-drug treatment. The longer the drug addiction lasts, the longer the anti-drug treatment takes.

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IJERPH, Vol. 13, Pages 176: Interactions of Six SNPs in ABCA1gene and Obesity in Low HDL-C Disease in Kazakh of China

Objective: To detect the interactions between six functional polymorphisms in ABCA1 and obesity in Kazakhs with low HDL-C levels. Methods: A total of 204 patients with low HDL-C and 207 health control subjects, which were randomly selected from among 5692 adult Kazakhs, were matched for age and sex. We genotyped ABCA1 single nucleotide polymorphisms of rs2515602, rs3890182, rs2275542, rs2230806, rs1800976, and rs4149313. Results: (1) The genotypic and allelic frequencies of rs2515602, rs2230806 and rs4149313 were different between normal HDL-C and low HDL-C subjects, the genotypic frequency of rs2275542 was also different between normal HDL-C and low HDL-C subjects (p < 0.05); (2) the level of HDL-C (rs2515602 and rs2275542) in normal HDL-C subjects were different among the genotypes (p < 0.05); the levels of TC, LDL-C (rs2515602, rs4149313); TG (rs2515602, rs1800976, rs4149313) in low HDL-C patients were different among the genotypes (p < 0.05); (3) interactions between the rs3890182, rs2275542, rs180096, and rs4149313 polymorphisms in ABCA1 gene and obesity may be associated with low HDL-C disease; (4) the C-C-C-A-A-G, T-C-C-A-A-A, T-C-C-A-A-G, C-C-C-A-A-A, C-T-G-G-A-A, and T-T-C-G-A-A haplotypes were significant between the subjects with normal HDL-C and low HDL-C level (p < 0.05). Conclusions: The differences in serum lipid levels between normal HDL-C and low HDL-C subjects among Kazakhs might partly result from ABCA1 gene polymorphisms; ABCA1 gene polymorphisms may be associated with low HDL-C disease; the low HDL-C disease might partly result from interactions between ABCA1 gene polymorphisms and obesity; the C-C-C-A-A-G, T-C-C-A-A-A, and T-C-C-A-A-G haplotypes may serve as risk factors of low HDL-C disease among Kazakhs, the C-C-C-A-A-A, C-T-G-G-A-A, and T-T-C-G-A-A haplotypes may serve as protective factor of low HDL-C disease among Kazakhs.

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IJERPH, Vol. 13, Pages 165: Environmental Change in the Agro-Pastoral Transitional Zone, Northern China: Patterns, Drivers, and Implications

Chengde city is located in the agro–pastoral transitional zone in northern China near the capital city of Beijing, which has experienced large-scale ecological construction in the past three decades. This study quantitatively assessed the environmental changes in Chengde through observation records of water resources, water environment, atmospheric environment, and vegetation activity and investigated the possible causes. From the late 1950s to 2002, the streamflow presented a downward trend induced by climate variability and human activities, with contribution ratios of 33.2% and 66.8%, respectively. During 2001–2012, the days of levels I and II air quality presented clear upward trends. Moreover, the air pollutant concentration was relatively low compared with that in the adjacent areas, which means the air quality has improved more than that in the neighboring areas. The water quality, which deteriorated during 1993–2000, began to improve in 2002. The air and water quality changes were closely related to pollutant emissions induced by anthropogenic activities. During 1982–2012, the vegetation in the southeastern and central regions presented restoration trends, whereas that in the northwestern area showed degradation trends. The pixels with obvious degradation trends correlated significantly with annual mean temperature and annual precipitation. Ecological engineering also played a positive role in vegetation restoration. This analysis can be beneficial to environment managers in the active response and adaptation to the possible effects of future climate change, population growth, and industrial development and can be used to ensure sustainable development and environmental safety.

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IJMS, Vol. 17, Pages 169: Icaritin Inhibits Collagen Degradation-Related Factors and Facilitates Collagen Accumulation in Atherosclerotic Lesions: A Potential Action for Plaque Stabilization

Most acute coronary syndromes result from rupture of vulnerable atherosclerotic plaques. The collagen content of plaques may critically affect plaque stability. This study tested whether Icaritin (ICT), an intestinal metabolite of Epimedium-derived flavonoids, could alter the collagen synthesis/degradation balance in atherosclerotic lesions. Rabbits were fed with an atherogenic diet for four months. Oral administration of ICT (10 mg·kg−1·day−1) was started after two months of an atherogenic diet and lasted for two months. The collagen degradation-related parameters, including macrophages accumulation, content and activity of interstitial collagenase-1 (MMP-1), and the collagen synthesis-related parameters, including amount and distribution of smooth muscle cells (SMC) and collagen mRNA/protein levels, were evaluated in the aorta. ICT reduced plasma lipid levels, inhibited macrophage accumulation, lowered MMP-1 mRNA and protein expression, and suppressed proteolytic activity of pro-MMP-1 and MMP-1 in the aorta. ICT changed the distribution of the SMCs towards the fibrous cap of lesions without increasing the amount of SMCs. Higher collagen protein content in lesions and aorta homogenates was observed with ICT treatment compared with the atherogenic diet only, without altered collagen mRNA level. These results suggest that ICT could inhibit the collagen degradation-related factors and facilitate collagen accumulation in atherosclerotic lesions, indicating a new potential of ICT in atherosclerotic plaques.

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IJMS, Vol. 17, Pages 168: Lingual Epithelial Stem Cells and Organoid Culture of Them

As tongue cancer is one of the major malignant cancers in the world, understanding the mechanism of maintenance of lingual epithelial tissue, which is known to be the origin of tongue cancer, is unquestionably important. However, the actual stem cells that are responsible for the long-term maintenance of the lingual epithelium have not been identified. Moreover, a simple and convenient culture method for lingual epithelial stem cells has not yet been established. Recently, we have shown that Bmi1-positive cells, residing at the second or third layer of the epithelial cell layer at the base of the interpapillary pit (IPP), were slow-cycling and could supply keratinized epithelial cells for over one year, indicating that Bmi1-positive cells are long-term lingual epithelial stem cells. In addition, we have developed a novel lingual epithelium organoid culture system using a three-dimensional matrix and growth factors. Here, we discuss current progress in the identification of lingual stem cells and future applications of the lingual culture system for studying the regulatory mechanisms of the lingual epithelium and for regenerative medicine.

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IJMS, Vol. 17, Pages 167: Mitotic Diversity in Homeostatic Human Interfollicular Epidermis

Despite decades of skin research, regulation of proliferation and homeostasis in human epidermis is still insufficiently understood. To address the role of mitoses in tissue regulation, we utilized human long-term skin equivalents and systematically assessed mitoses during early epidermal development and long-term epidermal regeneration. We now demonstrate four different orientations: (1) horizontal, i.e., parallel to the basement membrane (BM) and suggestive of symmetric divisions; (2) oblique with an angle of 45°–70°; or (3) perpendicular, suggestive of asymmetric division. In addition, we demonstrate a fourth substantial fraction of suprabasal mitoses, many of which are committed to differentiation (Keratin K10-positive). As verified also for normal human skin, this spatial mitotic organization is part of the regulatory program of human epidermal tissue homeostasis. As a potential marker for asymmetric division, we investigated for Numb and found that it was evenly spread in almost all undifferentiated keratinocytes, but indeed asymmetrically distributed in some mitoses and particularly frequent under differentiation-repressing low-calcium conditions. Numb deletion (stable knockdown by CRISPR/Cas9), however, did not affect proliferation, neither in a three-day follow up study by life cell imaging nor during a 14-day culture period, suggesting that Numb is not essential for the general control of keratinocyte division.

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Prevalence and antimicrobial susceptibility profiles of Staphylococcus aureus nasal carriage among pre-clinical and clinical medical students in a Tanzanian University

Methicillin resistant Staphylococcus aureus (MRSA) nasal carriage is a potential niche for spread and a risk factor for subsequent infections. Despite the fact that medical students are exposed to patients in the...

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A descriptive investigation of the impact of student research projects arising from elective research courses

Pharmacy academicians have noted the need to develop research skills in student pharmacists. At the Gatton College of Pharmacy, significant focus has been placed on the development of research skills through o...

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Toxicological study of the butanol fractionated root extract of Asparagus africanus Lam., on some blood parameter and histopathology of liver and kidney in mice

The butanol fractionated root extract of Asparagus africanus Lam., a traditional herb widely used to treat various ailments were analyzed for the presence of potential toxicity after single (acute) and repeated (...

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Formulation and Characterization of Bovine Serum Albumin-Loaded Niosome

Abstract

Niosomal vesicle, as a unique novel drug delivery system, is synthesized by non-ionic surfactants. Both hydrophilic and lipophilic drugs and also biomacromolecular agents, such as peptides and proteins can be encapsulated in this vesicular particle. Regarding polypeptide-based component loading, and delivery potential of the niosome, some valuable studies have been conducted in recent years. However, exploring the full potential of this approach requires fine tuned optimization and characterization approaches. Therefore, this study was conducted to achieve the following two goals. First, formulation and optimization of bovine serum albumin (BSA) load and release behavior as a function of cholesterol (CH) to sorbitan monostearate (Span 60) molar ratio. Second, investigating a cost- and time-effective polypeptide detecting method via methyl orange (MO) dye. To this aim, BSA-loaded niosomes were prepared by reversed-phase evaporation technique. The effect of CH to Sorbitan monostearate (Span 60) molar ratio on noisome entrapment efficiency (EE%) and release profile of BSA was studied using a ultraviolet (UV) spectrophotometer technique (NanoDrop 2000/2000c).Niosome with a 60% CH content showed the highest BSA EE% and release behavior. Then, BSA was dyed using MO in an acidic solution and used in BSA-niosome formulation. The MO-colored protein, loaded into the vesicles, was successfully assessed by an inverted light microscope, in order to observe the protein location in the vesicle. The results obtained in this study can be useful for various applications in different fields, including pharmaceutical, cosmetics, and drug delivery in biomedical and tissue engineering.



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Fusion Method for Solubility and Dissolution Rate Enhancement of Ibuprofen Using Block Copolymer Poloxamer 407

ABSTRACT

Aim of current research was to prepare ibuprofen-poloxamer 407 binary mixtures using fusion method and characterize them for their physicochemical and performance properties. Binary mixtures of ibuprofen and poloxamer were prepared in three different ratios (1:0.25, 1:0.5, and 1:0.75, respectively) using a water-jacketed high shear mixer. In vitro dissolution and saturation solubility studies were carried out for the drug, physical mixtures, and formulations for all ratios in de-ionized water, 0.1 N HCl (pH = 1.2), and phosphate buffer (pH = 7.2). Thermal and physical characterization of samples was done using modulated differential scanning calorimetry (mDSC), X-ray powder diffraction (XRD), and infrared spectroscopy (FTIR). Flow properties were evaluated using a powder rheometer. Maximum solubility enhancement was seen in acidic media for fused formulations where the ratio 1:0.75 had 18-fold increase. In vitro dissolution studies showed dissolution rate enhancement for physical mixtures and the formulations in all three media. The most pronounced effect was seen for formulation (1:0.75) in acidic media where the cumulative drug release was 58.27% while for drug, it was 3.67%. Model independent statistical methods and ANOVA based methods were used to check the significance of difference in the dissolution profiles. Thermograms from mDSC showed a characteristic peak for all formulations with Tpeak of around 45°C which suggested formation of a eutectic mixture. XRD data displayed that crystalline nature of ibuprofen was intact in the formulations. This work shows the effect of eutectic formation and micellar solubilization between ibuprofen and poloxamer at the given ratios on its solubility and dissolution rate enhancement.



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Luftemboli via intraossøs nål?

På sykehuset fikk mannen i 60-årene flere venekanyler i overekstremitetene. Den intraossøse nålen ble fjernet etter å ha vært åpen rundt en halvtime mens pasienten lå i sjokkleie.

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Double-sided fresnel zone plates as high performance optics in X-ray microscopy

X-ray microscopy describes a range of analytical techniques, specialized for the characterization of organic and inorganic samples using high energy photons. It takes advantage of the high penetration depth, high resolution and chemical sensitivity of X-rays and allows for the study of extended samples in their native environment without extensive sample preparation. Many of these experimental methods employ diffractive X-ray optics, like Fresnel zone plate lenses to obtain high spatial resolution or the better utilization of the incoming flux. Since improving the efficiency of zone plates can increase the throughput, quality and resolution of measurements, there is a constant demand for high efficiency and high resolution X-ray optics. Stacking is an established concept for extending the capabilities of zone plate optics. By stacking two zone plates in each other's optical near field, they act as a single zone plate with combined optical transmission profile, that would be infeasible to make as a single optical element. Yet the existing implementations of stacking suffer from issues regarding complexity and stability. This work presents the development of an alternative solution to conventional zone plate stacking, that circumvents most of its drawbacks. By patterning two zone plates on the front and back sides of a membrane, double-sided zone plates can deliver the advantages of stacked zone plates as inherently monolithic, single-chip optical elements. Double-sided blazed zone plates with two complementary binary zone plates on the two sides of the membrane were produced to provide an effective four level transmission profile. This allowed to bypass the fundamental limitations of binary zone plates by providing up to 54.7% diffraction efficiency at 6.2 keV while having 200 nm smallest half-pitch and a reasonable working distance. For high resolution zone plates, structure height is the main limiting factor. Therefore by patterning two identical zone plates on the two sides of the membrane, one can double the effective structure height. This provided us with a significant gain in focusing efficiency at high photon energies, as we have successfully measured 9.9% focusing efficiency at 9 keV with 30 nm smallest half-pitch, while preserving diffraction limited optical performance. Stacking two complementary zone plates for multiplying their spatial frequencies opens the possibility for ultra-high resolution zone plate optics. We have successfully produced and tested interlaced zone plate optics down to 7 nm smallest half-pitch while still maintaining practical aperture sizes. This thesis is a comprehensive summary of the work performed for the fabrication and characterization of the high performance zone plates representing each concept and provides possible examples for their future use.

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Impaired topological architecture of brain structural networks in idiopathic Parkinson’s disease: a DTI study

Abstract

Parkinson’s disease (PD) is considered as a neurodegenerative disorder of the brain central nervous system. But, to date, few studies adopted the network model to reveal topological changes in brain structural networks in PD patients. Additionally, although the concept of rich club organization has been widely used to study brain networks in various brain disorders, there is no study to report the changed rich club organization of brain networks in PD patients. Thus, we collected diffusion tensor imaging (DTI) data from 35 PD patients and 26 healthy controls and adopted deterministic tractography to construct brain structural networks. During the network analysis, we calculated their topological properties, and built the rich club organization of brain structural networks for both subject groups. By comparing the between-group differences in topological properties and rich club organizations, we found that the connectivity strength of the feeder and local connections are lower in PD patients compared to those of the healthy controls. Furthermore, using a network-based statistic (NBS) approach, we identified uniformly significantly decreased connections in two modules, the limbic/paralimbic/subcortical module and the cognitive control/attention module, in patients compared to controls. In addition, for the topological properties of brain network topology in the PD patients, we found statistically increased shortest path length and decreased global efficiency. Statistical comparisons of nodal properties were also widespread in the frontal and parietal regions for the PD patients. These findings may provide useful information to better understand the abnormalities of brain structural networks in PD patients.



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Soil Influences Colonization of Root-Associated Fungal Endophyte Communities of Maize, Wheat, and Their Progenitors

Root-associated fungal endophytes are vital component of root microbiome as some mitigate their host’s abiotic and biotic stress. We characterized root-associated fungal endophytes in cereal grains and their progenitors grown on two different soil-types. We aimed at determining how clay and desert soil affects the colonization of root fungal community. Both culture-dependent and culture-independent methods were employed to identify endophytes that successfully colonized greenhouse-grown host plants. The Internal Transcriber Spacer region of fungal ribosomal DNA was utilized for identification purposes. This study revealed soil as a prominent factor influencing the composition of microfungal communities inhabiting the roots of maize (Zea mays subsp. mays) and its conspecific progenitor, teosinte (Zea mays subsp. parviglumis). Similar results were found in wheat (Triticum aestivum subsp. aestivum) and its progenitor (Triticum monococcum subsp. monococcum). The multidimensional comparisons of Morisita-Horn similarity values of fungal colonists of various host plant taxa indicated that soil plays a primary role in shaping the root fungal community; a secondary effect was plant host identity, even when the plant host is a conspecific. Future studies focused on characterizing root endophytes in other cereal grains, and studying the effect of edaphic factors on fungal colonization, can ultimately contribute to crop productivity.

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Postobstructive Pulmonary Edema following Tonsillectomy/Adenoidectomy in a 2-Year-Old with Poland-Moebius Syndrome

A 2-year-old male with Poland-Moebius syndrome was transferred from a local hospital to the Pediatric ICU at Children’s Hospital of Georgia for suspected postobstructive pulmonary edema (POPE) after tonsillectomy/adenoidectomy (T&A). The patient’s respiratory status ultimately declined and he developed respiratory failure. Imaging suggested pulmonary edema as well as a left-sided pneumonia. Echocardiogram showed pulmonary hypertension and airway exam via direct fiberoptic bronchoscopy revealed tracheomalacia and bronchomalacia. He developed acute respiratory distress syndrome (ARDS) and remained intubated for ten days. This case highlights the association between congenital upper body abnormalities with cranial nerve dysfunction and the development of POPE with delayed resolution of symptoms. Patients with upper body abnormalities as above are at great risk of postoperative complications and should therefore be managed in a tertiary-care facility.

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Xyloglucan Based In Situ Gel of Lidocaine HCl for the Treatment of Periodontosis

The present study was aimed at formulating thermoreversible in situ gel of local anesthetic by using xyloglucan based mucoadhesive tamarind seed polysaccharide (TSP) into periodontal pocket. Temperature-sensitive in situ gel of lidocaine hydrochloride (LH) (2% w/v) was formulated by cold method. A full 32 factorial design was employed to study the effect of independent variables concentrations of Lutrol F127 and TSP to optimize in situ gel. The dependent variables evaluated were gelation temperature () and drug release (). The results revealed the surface pH of 6.8, similar to the pH of saliva. Viscosity study showed the marked increase in the viscosity of gel at 37°C due to sol-gel conversion. TSP was found to act as good mucoadhesive component to retain gel at the site of application in dental pocket. Gelation of formulation occurred near to body temperature. In vitro study depicted the fast onset of drug action but lasting the release (90%) till 2 h. Formulation F7 was considered as optimized batch, containing 18% Lutrol F127 and 1% tamarind seed polysaccharide. Thus, lidocaine hydrochloride thermoreversible in situ gel offered an alternative to painful injection therapy of anesthesia during dental surgery, with fast onset of anesthetic action lasting throughout the dental procedure.

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Restoration of the Ellipsoid Zone and Visual Prognosis at 1 Year after Surgical Macular Hole Closure

Purpose. To evaluate the restoration of the ellipsoid zone (EZ) and its influence on visual prognosis 1 year after surgical macular hole (MH) closure. Method. Subjects were patients with stage 2, 3, or 4 idiopathic MH who underwent primary vitrectomy that resulted in successful hole closure. Nineteen eyes with both EZ disruption with foveal detachment and a continuous external limiting membrane on optical coherence tomography during the early postoperative period were included in this study. Result. EZ disruption was restored in 10 eyes (53%, Group A) and remained in 9 eyes (47%, Group B) at 1 year after surgery. In Group B, the diameter of the residual EZ disruption was  μm. LogMAR visual acuity (VA) 1 year after surgery was significantly better than preoperative VA in each group (Group A: ; ; Group B: ; ), but there was no significant difference between the 2 groups (). There was no significant correlation between logMAR VA and EZ disruption diameter at 1 year after surgery. Conclusion. EZ was restored in 53% of eyes at 1 year after surgical closure of idiopathic MH. Mean residual EZ disruption diameter was  μm. Neither resolved nor residual EZ disruption influenced postoperative VA.

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Preclinical Models for Investigation of Herbal Medicines in Liver Diseases: Update and Perspective

Liver disease results from a dynamic pathological process associated with cellular and genetic alterations, which may progress stepwise to liver dysfunction. Commonly, liver disease begins with hepatocyte injury, followed by persistent episodes of cellular regeneration, inflammation, and hepatocyte death that may ultimately lead to nonreversible liver failure. For centuries, herbal remedies have been used for a variety of liver diseases and recent studies have identified the active compounds that may interact with liver disease-associated targets. Further study on the herbal remedies may lead to the formulation of next generation medicines with hepatoprotective, antifibrotic, and anticancer properties. Still, the pharmacological actions of vast majority of herbal remedies remain unknown; thus, extensive preclinical studies are important. In this review, we summarize progress made over the last five years of the most commonly used preclinical models of liver diseases that are used to screen for curative herbal medicines for nonalcoholic fatty liver disease, liver fibrosis/cirrhosis, and liver. We also summarize the proposed mechanisms associated with the observed liver-protective, antifibrotic, and anticancer actions of several promising herbal medicines and discuss the challenges faced in this research field.

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Circulating Carbonic Anhydrase IX and Antiangiogenic Therapy in Breast Cancer

Introduction. Carbonic anhydrase IX (CAIX) is a hypoxia regulated metalloenzyme integral to maintaining cellular pH. Increased CAIX expression is associated with poor prognosis in breast cancer. To explore CAIX as a biomarker for breast cancer therapies, we measured plasma CAIX levels in healthy control subjects and in breast cancer patients. Methods. In control subjects we evaluated plasma CAIX stability via commercially available ELISA. We then similarly quantified plasma CAIX levels in (1) locally advanced breast cancer (LABC) patients treated with neoadjuvant paclitaxel + sunitinib (T + S) followed by doxorubicin and cyclophosphamide (AC); (2) metastatic breast cancer (MBC) patients treated with systemic chemotherapy. Results. Plasma CAIX levels were stable at room temperature for at least 48 hours in control subjects. Mean baseline plasma CAIX levels were lower in controls compared to patients with LABC or MBC. In LABC, CAIX levels rose significantly in response to administration of antiangiogenic therapy (T + S) () but not AC (). In patients with MBC treated without an antiangiogenic agent CAIX levels did not change with therapy. Conclusions. Our results suggest that CAIX may be an easily obtained, stable measure of tumor associated hypoxia as well as a useful pharmacodynamic biomarker for antiangiogenic therapy.

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ChemTok: A New Rule Based Tokenizer for Chemical Named Entity Recognition

Named Entity Recognition (NER) from text constitutes the first step in many text mining applications. The most important preliminary step for NER systems using machine learning approaches is tokenization where raw text is segmented into tokens. This study proposes an enhanced rule based tokenizer, ChemTok, which utilizes rules extracted mainly from the train data set. The main novelty of ChemTok is the use of the extracted rules in order to merge the tokens split in the previous steps, thus producing longer and more discriminative tokens. ChemTok is compared to the tokenization methods utilized by ChemSpot and tmChem. Support Vector Machines and Conditional Random Fields are employed as the learning algorithms. The experimental results show that the classifiers trained on the output of ChemTok outperforms all classifiers trained on the output of the other two tokenizers in terms of classification performance, and the number of incorrectly segmented entities.

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DVD Review



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Issue Information



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NMR Concepts



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Oncogenic activation of the PI3K/Akt pathway promotes cellular glucose uptake by downregulating the expression of thioredoxin-interacting protein

Publication date: Available online 28 January 2016
Source:Cellular Signalling
Author(s): Shin Yee Hong, Fa-Xing Yu, Yan Luo, Thilo Hagen
Oncogenic activation of the PI3K/Akt pathway is known to play an important role in promoting glucose metabolism in cancer cells. However, the molecular mechanism through which the PI3K/Akt signalling pathway promotes glucose utilisation in cancer cells is still not well understood. It has recently been shown that the oncogenic activation of the PI3K/Akt/mTOR signalling in lung adenocarcinoma is important in promoting the localisation of glucose transporter 1 (GLUT1) at the plasma membrane. We thus hypothesised that the effect of constitutive activation of the PI3K/AKT signalling on glucose metabolism is mediated by thioredoxin interacting protein (TXNIP), a known regulator of the GLUT1 plasma membrane localisation. Consistent with previous studies, inhibition of the PI3K/Akt pathway decreased cellular glucose uptake. Furthermore, inhibition of PI3K/Akt signalling in non-small cell lung cancer (NSCLC) cell lines using clinically used tyrosine kinase inhibitors (TKIs) resulted in a decrease in GLUT1 membrane localisation. We also observed that inhibition of the PI3K/Akt pathway in various cell lines, including NSCLC cells, resulted in an increase in TXNIP expression. Importantly, knockdown of TXNIP using siRNA in the NSCLC cells promoted GLUT1 to be localised at the plasma membrane and reversed the effect of PI3K/Akt inhibitors. Together, our results suggest that the oncogenic activation of PI3K/Akt signalling promotes cellular glucose uptake, at least in part, through the regulation of TXNIP expression. This mechanism may contribute to the Warburg effect in cancer cells.



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Transforming growth factor type beta (TGF- β) requires reactive oxygen species to induce skeletal muscle atrophy

Publication date: Available online 26 January 2016
Source:Cellular Signalling
Author(s): Johanna Abrigo, Juan Carlos Rivera, Felipe Simon, Daniel Cabrera, Claudio Cabello-Verrugio
Transforming growth factor beta 1 (TGF- β1) is a classical modulator of skeletal muscle and regulates several processes, such as myogenesis, regeneration, and muscle function in skeletal muscle diseases. Skeletal muscle atrophy, characterized by the loss of muscle strength and mass, is one of the pathological conditions regulated by TGF- β. Atrophy also results in increased myosin heavy chain (MHC) degradation and the expression of two muscle-specific E3 ubiquitin ligases, atrogin-1 and MuRF-1. Reactive oxygen species (ROS) are modulators of muscle wasting, and NAD(P)H oxidase (NOX) is one of the main sources of ROS. While it was recently found that TGF- β1 induces atrophy in skeletal muscle, the underlying mechanism is not fully understood. In this study, the role of NOX-derived ROS in skeletal muscle atrophy induced by TGF- β was assessed. TGF- β1 induced an atrophic effect in C2C12 myotubes, as evidenced by decreased myotube diameter and MHC levels, together with increased MuRF-1 levels. Concomitantly, TGF- β increased NOX-induced ROS contents. Interestingly, NOX inhibition through apocynin and the antioxidant treatment with N-acetyl cysteine (NAC) decreased increased ROS levels in myotubes. Additionally, both apocynin and NAC completely prevented the decreased MHC, decreased myotube diameter, and increased MuRF-1 induced by TGF-β. Injection of TGF- β1 into the tibialis anterior muscle induced atrophy, as observed by decreased fibre diameter and MHC levels, together with increased MuRF-1 levels. Likewise, TGF-β increased the ROS contents in the smaller fibres of skeletal muscle. Additionally, the administration of NAC to mice prevented all atrophic effects and the increase in ROS induced by TGF-β in the tibialis anterior. This is the first study to report that TGF-β has an atrophic effect dependent on NOX-induced ROS in skeletal muscle.



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Analysis of the antibacterial effect of an Edwardsiella tarda LuxS inhibitor

LuxS/AI-2 quorum sensing is involved in the virulence of many bacterial pathogens, including the fish pathogen Edwardsiella tarda. In a previous study, we identified a small peptide, 5906, which inhibits E. tarda LuxS activity by binding specifically to LuxS in a manner that probably prevents the formation of functional LuxS homodimer. In the present study, using Japanese flounder as the experimental animal, we analyzed the antibacterial effect of 5906 produced by DH5α/p5906 (an Escherichia coli strain that produces 5906) and pID5906 (a mammalian plasmid that functional in flounder constitutively expresses 5906) against different bacterial fish pathogens. We found that fish administered with both DH5α/p5906 and pID5906 exhibited reduced bacterial recovery following E. tarda challenge. We also examined the effect of 5906 on the infection caused by another two fish pathogen, Aeromonas hydrophila and Vibrio harveyi. The results indicated that 5906 produced by DH5α/p5906 inhibited the AI-2 activity of A. hydrophila and V. harveyi, and that fish administered with DH5α/p5906 showed enhanced resistance against challenges with both bacteria. These results suggest that 5906 or its analogues/derivatives may be exploited for the development of broad-spectrum antibacterial agents applied in the prevention and control of fish bacterial diseases.

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Differential pain response at local and remote muscle sites following aerobic cycling exercise at mild and moderate intensity

Physical exercise has been shown to inhibit experimental pain response in the post-exercise period. Modulation of the pain system may be differentiated between muscle sites engaging in contractile activity. The purpose of this study was to assess the pain response at remote and local muscle sites following aerobic exercise at different work intensities. Participants included 10 healthy and physically active males (mean age ± SD, 21.2 ± 3.4). Somatic pressure pain threshold (PPT) at the rectus femoris (local) and brachioradialis (remote) muscle site was measured at before (Pre), 5 min after (Post1), and 15 min after (Post2) aerobic cycling exercise at 70 and 30 % of peak oxygen uptake (VO 2peak ) performed on different occasions in a counterbalanced order, separated by minimum of 3 days interval. Repeated measures ANOVA for PPT reveals significant main effect for time (f = 3.581, p = 0.049, observed power = 0.588) and muscle site (f = 17.931, p = 0.002, observed power = 0.963). There was a significant interaction shown for exercise intensity by time (f = 11.390, p = 0.012, observed power = 0.790). PPT at rectus femoris following cycling exercise at 70 % of VO 2peak reveals a significant increase between Pre-Post1 (p = 0.040). PPT for rectus femoris following cycling exercise at 30 % of VO 2peak revealed a significant decrease between Pre-Post1 (p = 0.026) and Pre-Post2 (p = 0.008). The PPT for brachioradialis following cycling exercise at 30 % of VO 2peak revealed a significant decrease between Pre-Post1 (p = 0.011) and Pre-Post2 (p = 0.005). These results show that aerobic exercise increases PPT locally at the exercise muscle site following exercise at 70 % of VO 2peak but reduces PPT following exercise at 30 % of VO 2peak .

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Value of Radiofrequency Ablation in The Management of Retropharyngeal Lymphatic Malformation

Lymphatic malformations are benign malformations frequently occurring in the head and neck. Retropharyngeal location is rare, can be life threating and its management is particularly challenging. Over a three-year period, three patients presented with symptomatic (dyspnea and/or dysphagia) retropharyngeal lymphatic malformation. All were treated using a radiofrequency ablation of lymphatic malformation through a trans-oral approach. No major complications occurred following the surgery. During the follow-up, no recurrence was noted and all patients were asymptomatic.

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Production and evaluation of a Spanish version of the LittlEARS® Auditory Questionnaire for the assessment of auditory development in children

To adapt the LittlEARS® Auditory Questionnaire (LEAQ) into Spanish and evaluate the psychometric properties of the Spanish version of the questionnaire.

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The Association of Congenital Tracheobronchial Stenosis and Cardiovascular Anomalies

Congenital tracheobronchial stenosis (CTBS) is a rare disorder characterized by the presence of focal or diffuse complete tracheal or bronchial cartilage rings resulting in a fixed lumen narrowing. The aim of this study was to expose the association of various cardiovascular (CV) anomalies with various types of CTBS.

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Treatment of Severe Tracheobronchomalacia: Ten-year experience

Paediatric tracheobronchomalacia is a rare but potentially serious condition. Severe tracheobronchomalacia requires intervention or operation. This is an evaluation of a ten-year experience at an institution.

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Polysomnography results in pediatric patients with mild obstructive sleep apnea: Adenotonsillectomy vs. watchful waiting

There is a lack of consensus and a paucity of data regarding how to best treat pediatric patients with mild obstructive sleep apnea. The objective of our study was to compare outcomes following adenotonsillectomy versus observation in children with mild obstructive sleep apnea based on polysomnography results.

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polymorphisms and susceptibility to recurrent tonsillitis in Italian children

the tonsils are secondary lymphoid organs fundamental for immune system response against pathogens within the oral cavity. Tonsillitis refers to inflammation of the pharyngeal tonsils that may include the adenoids and the lingual tonsils and that can be acute, recurrent, and chronic. Viral or bacterial infections, as well as immunologic factors are the main trigger to tonsillitis and disease's chronicity: the host immune responses, especially the innate one, could play an important role in susceptibility to the disease.

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Suicide attempt with a mix of synthetic cannabinoids and synthetic cathinones: Case report of non-fatal intoxication with AB-CHMINACA, AB-FUBINACA, alpha-PHP, alpha-PVP and 4-CMC

The use of designer drugs has risen severely in recent years. Among others, synthetic cannabinoids and synthetic cathinones have been recognized as the largest categories of new drugs identified in Europe last year [1]. Synthetic cannabinoids are novel psychoactive substances that imitate the effects of cannabis, while synthetic cathinones belong to phenylalkylamine group of drugs with characteristics similar to amphetamines. Their production appears to take place primarily in China and India. The drugs are then imported into Europe, where they are packaged and marketed as ‘legal highs’ or sold in the illicit market [1,2].

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The effects of hemoglobin levels and their interactions with cigarette smoking on survival in nasopharyngeal carcinoma patients

Abstract

There is very little published information regarding the prognostic value of hemoglobin (Hb) levels combined with smoking on the survival of patients with nasopharyngeal carcinoma (NPC), and the interactions between them remain unclear. A total of 2440 NPC patients were confirmed, and multivariate analysis was performed to identify valuable prognostic Hb levels in the entire population and in the cohort of smokers. The survival differences were compared using log-rank tests. The multiplicative and additive interactions were assessed using Cox regression and a Microsoft Word Excel spreadsheet. Postradiotherapy (RT) Hb was an independent prognostic factor for overall survival (OS) (HR = 0.797; = 0.006), failure-free survival (FFS) (HR=0.811; = 0.010), and loco-regional failure-free survival (LR-FFS) (HR = 0.725; = 0.000). In the cohort of smokers, pack-years was also an independent predictor of OS (HR = 0.673; < 0.001) and FFS (HR = 0.681; < 0.001), LR-FFS (HR = 0.663; = 0.001). A significant positive additive effect was found for the interaction between low post-RT Hb and high SI on OS, with RERI = 5.616, AP = 0.665, and S = 4.078. Stratified analyses demonstrated that heavy smokers with low post-RT Hb had HRs of 2.295 (< 0.001) for death, 2.222 (< 0.001) for disease failure, and 2.267 (< 0.001) loco-regional recurrence compared with light smokers with high post-RT Hb levels, and post-RT Hb level is an important predictor of survival in patients with NPC. The positive interaction between post-RT Hb level and pack-years contributes to the elevated risk of poor survival. Oncologists should devote particular attention to heavy smokers with low post-RT Hb levels in the future.

Thumbnail image of graphical abstract

Post-RT Hb level is an important predictor of survival in patients with nasopharyngeal carcinoma (NPC). Smoking index is also an independent predictor of survival in smoking NPC patients. The positive interaction between post-RT Hb level and smoking index contribute to an elevated risk of poor survival.



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Children's comprehension of an unfamiliar speaker accent: a review

Abstract

Background

The effect of speaker accent on listeners’ comprehension has become a key focus of research given the increasing cultural diversity of society and the increased likelihood of an individual encountering a clinician with an unfamiliar accent.

Aims

To review the studies exploring the effect of an unfamiliar accent on language comprehension in typically developing (TD) children and in children with speech and language difficulties. This review provides a methodological analysis of the relevant studies by exploring the challenges facing this field of research and highlighting the current gaps in the literature.

Methods & Procedures

A total of nine studies were identified using a systematic search and organized under studies investigating the effect of speaker accent on language comprehension in (1) TD children and (2) children with speech and/or language difficulties.

Main Contribution

This review synthesizes the evidence that an unfamiliar speaker accent may lead to a breakdown in language comprehension in TD children and in children with speech difficulties. Moreover, it exposes the inconsistencies found in this field of research and highlights the lack of studies investigating the effect of speaker accent in children with language deficits.

Conclusions & Implications

Overall, research points towards a developmental trend in children's ability to comprehend accent-related variations in speech. Vocabulary size, language exposure, exposure to different accents and adequate processing resources (e.g. attention) seem to play a key role in children's ability to understand unfamiliar accents. This review uncovered some inconsistencies in the literature that highlight the methodological issues that must be considered when conducting research in this field. It explores how such issues may be controlled in order to increase the validity and reliability of future research. Key clinical implications are also discussed.



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The role of IL-11 in immunity and cancer

• IL-11 mainly acts as an anti-inflammatory cytokine though it can function as a pro-inflammatory cytokine.• IL-11 promotes tumorigenesis by triggering the JAK-STAT3 pathway.

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Necrobiotic xanthogranuloma associated with monoclonal gammopathy: successful treatment with lenalidomide and dexamethasone



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Progesterone modulates endothelial progenitor cell (EPC) viability through the CXCL12/CXCR4/PI3K/Akt signalling pathway

Abstract

Objectives

Progesterone treatment can effectively increase levels of circulating endothelial progenitor cells (EPCs) and improve neurological functional outcome in a traumatic brain injury (TBI) rat model. However, the mechanisms of progesterone's effects on EPC viability remain elusive. The CXCL12/CXCR4 (CXC chemokine ligand 12/CXC chemokine receptor 4) signalling pathway regulates cell proliferation; we hypothesize that it mediates progesterone-induced EPC viability.

Materials and methods

EPCs were isolated from bone marrow-derived mononuclear cells (BM-MNCs) and treated with progesterone (5, 10 and 100 nm). MTS assay was used to investigate EPC viability. Protein expression was examined by Western blotting, ELISA assay and flow cytometry. Cell membrane and cytoplasm proteins were extracted with membrane and cytoplasm protein extraction kits. CXCR4 antagonist (AMD3100) and phosphatidylinositol 3-kinases (PI3K) antagonist (LY294002) were used to characterize underlying mechanisms.

Results

Progesterone-induced EPC viability was time- and dose-dependent. Administration of progesterone facilitated EPC viability and increased expression of CXCL12 and phosphorylated Akt (also known as protein kinase B, pAkt) activity (P < 0.05). Progesterone did not regulate CXCR4 protein expression in cultured EPC membranes or cytoplasm. However, progesterone-induced EPC viability was significantly attenuated by AMD3100 or LY294002. Inhibition of the signalling pathway with AMD3100 and LY294002 subsequently reduced progesterone-induced CXCL12/CXCR4/PI3K/pAkt signalling activity.

Conclusions

The CXCL12/CXCR4/PI3K/pAkt signalling pathway increased progesterone-induced EPC viability.



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Interplay between Cellular and Molecular Inflammatory Mediators in Lung Cancer

Inflammation is a component of the tumor microenvironment and represents the 7th hallmark of cancer. Chronic inflammation plays a critical role in tumorigenesis. Tumor infiltrating inflammatory cells mediate processes associated with progression, immune suppression, promotion of neoangiogenesis and lymphangiogenesis, remodeling of extracellular matrix, invasion and metastasis, and, lastly, the inhibition of vaccine-induced antitumor T cell response. Accumulating evidence indicates a critical role of myeloid cells in the pathophysiology of human cancers. In contrast to the well-characterized tumor-associated macrophages (TAMs), the significance of granulocytes in cancer has only recently begun to emerge with the characterization of tumor-associated neutrophils (TANs). Recent studies show the importance of CD47 in the interaction with macrophages inhibiting phagocytosis and promoting the migration of neutrophils, increasing inflammation which can lead to recurrence and progression in lung cancer. Currently, therapies are targeted towards blocking CD47 and enhancing macrophage-mediated phagocytosis. However, antibody-based therapies may have adverse effects that limit its use.

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Synergy between Rhizobium phaseoli and Acidithiobacillus ferrooxidans in the Bioleaching Process of Copper

This study investigates the synergy of Rhizobium phaseoli and Acidithiobacillus ferrooxidans in the bioleaching process of copper. The results showed that additional R. phaseoli could increase leaching rate and cell number of A. ferrooxidans. When the initial cell number ratio between A. ferrooxidans and R. phaseoli was 2 : 1, A. ferrooxidans attained the highest final cell number of approximately 2 × 108 cells/mL and the highest copper leaching rate of 29%, which is 7% higher than that in the group with A. ferrooxidans only. R. phaseoli may use metabolized polysaccharides from A. ferrooxidans, and organic acids could chelate or precipitate harmful heavy metals to reduce their damage on A. ferrooxidans and promote its growth. Organic acids could also damage the mineral lattice to increase the leaching effect.

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Quality Assurance Competition Strategy under B2C Platform

This paper examines the platform competition strategy in offering quality assurance policy based on two-sided B2C markets with buyers and sellers who trade through the B2C platform. We model this as a strategy decision whether control rights of quality assurance level are better held by the seller or by the platform. Whether the seller or the platform that provides quality assurance policy is preferred depends on their influence on network effects. We show that the seller and the platform have no strong intention to improve the quality assurance level in the monopoly markets. When all buyers multihome and all sellers single-home, sellers who hold the control rights of quality assurance are better than platforms. However, platforms that hold quality assurance drive more profits than sellers in the B2C markets where all buyers and all sellers multihome. Our findings connect platform competitive strategies to market and microfoundation of quality, which applies directly to managerial decisions in the B2C two-sided platform.

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Adaptive Randomized Ensemble Tracking Using Appearance Variation and Occlusion Estimation

Tracking-by-detection methods have been widely studied with promising results. These methods usually train a classifier or a pool of classifiers in an online manner and use previous tracking results to generate a new training set for object appearance and update the current model to predict the object location in subsequent frames. However, the updating process may easily cause drifting in terms of appearance variation and occlusion. The previous methods for updating the classifier(s) decided whether or not to update the classifier(s) by a fixed learning rate parameter in all scenarios. The learning rate parameter has a great influence on the tracker’s performance and should be dynamically adjusted according to the change of scene during tracking. In this paper, we propose a novel method to model the time-varying appearance of an object that takes appearance variation and occlusion of local patches into consideration. In contrast with the existing methods, the learning rate for updating classifier ensembles adaptively is adjusted by estimating the appearance variation with sparse optical flow and the possible occlusion of the object between consecutive frames. Experiments and evaluations on some challenging video sequences have been done and the results demonstrate that the proposed method is more robust against appearance variation and occlusion than those state-of-the-art approaches.

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Trastuzumab in Esophagogastric Cancer: HER2-Testing and Treatment Reality outside Clinical Studies in Germany

We analysed trends over time in palliative first-line chemotherapy in patients with locally advanced or metastatic esophagogastric cancer. Special focus was on frequency and quality of HER2-testing and trends in drug use in combination with trastuzumab. Earlier published data about patients treated outside clinical studies showed a relatively low rate of HER2-testing and insufficient test quality. A total of 2,808 patients retrospectively documented in Therapiemonitor from 2006 to 2013 were analysed regarding treatment intensity and trends in used drugs. Data on HER2-testing and therapies were analysed in two cohorts documented in 2010 and 2011 (1) compared to 2012 and 2013 (2). Treatment intensity increased: 49.3% of patients received at least a triplet in 2013 compared to 10.1% in 2006. In cohort 2 HER2 expression was tested in 79.1% of the cases. Still, in 26.9% testing was not done as requested by guidelines. Good performance status, multiple metastases, age ≤ 65 years, the objective “to prevent progression,” good cognitive capabilities, estimated good compliance, and social integration positively influenced the probability of HER2-testing; comorbidities negatively affected it. Usage of the combination of fluoropyrimidines and cisplatin with trastuzumab declined from 67% in cohort 1 to 50% in cohort 2.

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Mechanical and Histological Effects of Resorbable Blasting Media Surface Treatment on the Initial Stability of Orthodontic Mini-Implants

Introduction. This study aimed to evaluate the effects of resorbable blasting media (RBM) treatment on early stability of orthodontic mini-implants by mechanical, histomorphometric, and histological analyses. Methods. Ninety-six (64 for mechanical study and 32 for histological study and histomorphometric analysis) titanium orthodontic mini-implants (OMIs) with machined (machined group) or RBM-treated (CaP) surface (RBM group) were implanted in the tibiae of 24 rabbits. Maximum initial torque (MIT) was measured during insertion, and maximum removal torque (MRT) and removal angular momentum (RAM) were measured at 2 and 4 weeks after implantation. Bone-to-implant contact (BIC) and bone area (BA) were analyzed at 4 weeks after implantation. Results. RBM group exhibited significantly lower MIT and significantly higher MRT and RAM at 2 weeks than machined group. No significant difference in MRT, RAM, and BIC between the two groups was noted at 4 weeks, although BA was significantly higher in RBM group than in machined group. RBM group showed little bone resorption, whereas machined group showed new bone formation after bone resorption. Conclusions. RBM surface treatment can provide early stability of OMIs around 2 weeks after insertion, whereas stability of machined surface OMIs may decrease in early stages because of bone resorption, although it can subsequently recover by new bone apposition.

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Protective Effect of Diospyros kaki against Glucose-Oxygen-Serum Deprivation-Induced PC12 Cells Injury

Ischemic cerebrovascular disease is one of the most common causes of death in the world. Recent interests have been focused on natural antioxidants and anti-inflammatory agents as potentially useful neuroprotective agents. Diospyros kaki (persimmon) has been shown to exert anti-inflammatory, antioxidant, and antineoplastic effects. However, its effects on ischemic damage have not been evaluated. Here, we used an in vitro model of cerebral ischemia and studied the effects of hydroalcoholic extract of peel (PeHE) and fruit pulp (PuHE) of persimmon on cell viability and markers of oxidative damage mainly intracellular reactive oxygen species (ROS) induced by glucose-oxygen-serum deprivation (GOSD) in PC12 cells. GOSD for 6 h produced significant cell death which was accompanied by increased levels of ROS. Pretreatment with different concentrations of PeHE and PuHE (0–500 μg/mL) for 2 and 24 h markedly restored these changes only at high concentrations. However, no significant differences were seen in the protection against ischemic insult between different extracts and the time of exposure. The experimental results suggest that persimmon protects the PC12 cells from GOSD-induced injury via antioxidant mechanisms. Our findings might raise the possibility of potential therapeutic application of persimmon for managing cerebral ischemic and other neurodegenerative disorders.

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