Immunophenotype characteristics of peripheral blood mononuclear leukocytes of chronic idiopathic urticaria patients.
The pathogenesis of chronic idiopathic urticaria (CIU) is not completely understood although autoimmunity has been proposed. The aim of the study was to assess the expression of different leukocyte antigens, by flow cytometry, assaying total blood of 29 patients with CIU and of 20 sex and age matched controls. Moreover, we assessed soluble CD154 a marker of immune cell activation, predominantly memory T cells. When patients were divided depending an their response to the autologous serum skin test (ASST), three different groups were encountered: group 1 (n=11): with negative ASST-, group 2 (n=11): positive ASST (ASST+) with normal lymphocyte counts and group 3 (n=7): ASST+ with low lymphocyte counts (< 1500 cells/mm3). A significant increase in CD19+ percentage and not in the absolute count (P < 0.05) was observed in group 1 as compared to controls and to the other groups. In contrast, CD30+, CD45RO+ and CD4+/CD45RO+ percentages and biologically active soluble CD154 levels were significantly higher (P < 0.05) in group 3 as compared to group 1 or to controls. In ASST positive groups, CD45RO+ and CD4+/CD45RO+ positiveness correlates with wheal diameter. In conclusion, memory cells may play a role in these different types of patients and in understanding CIU pathogenesis.


Comparative evaluation of three methods for quantifying tablet brittleness.
A low value of deformation before crushing is an obvious and understandable measure of brittleness of materials including tablets. In this article, three methods based on deformation measurement in a flexure tester are compared. The simplest one is a plain measurement of distance from contact or selected start point till fracture. Next the brittle-ductile method (BDI), where the distance is established by normalisation of the force-displacement curve based on the work of failure (WOF). The third method is the tablet brittleness index (TBI) by Gong and Sun, where the reciprocal of a linear distance is proposed as a brittleness quantity. The study is based on data from a previous investigation, where tablets of microcrystalline cellulose and lactose in different combinations and with four different crushing forces were utilised. The investigation shows that the BDI method is preferable. It is easy to compute without data manipulation, the sensitivity to the fracture force is negligible and it provides an independent characteristic of the brittleness of a compacted material.


AZD5363 Inhibits Inflammatory Synergy between Interleukin-17 and Insulin/Insulin-Like Growth Factor 1.
In the United States, one-third of population is affected by obesity and almost 29 million people are suffering from type 2 diabetes. Obese people have elevated serum levels of insulin, insulin-like growth factor 1 (IGF1), and interleukin-17 (IL-17). Insulin and IGF1 are known to enhance IL-17-induced expression of inflammatory cytokines and chemokines, which may contribute to the chronic inflammatory status observed in obese people. We have previously demonstrated that insulin/IGF1 signaling pathway crosstalks with IL-17-activated nuclear factor-κB pathway through inhibiting glycogen synthase kinase 3β (GSK3β) activity. However, it is unclear whether GSK3α also plays a role and whether this crosstalk can be manipulated by AZD5363, a novel pan-Akt inhibitor that has been shown to increase glycogen synthase kinase 3 activity through reducing phosphorylation of GSK3α and GSK3β. In this study, we investigated IL-17-induced expression of C-X-C motif ligand 1 (Cxcl1), C-C motif ligand 20 (Ccl20), and interleukin-6 (Il-6) in wild-type, GSK3α(-/-), and GSK3β(-/-) mouse embryonic fibroblast cells as well as in mouse prostate tissues by real-time quantitative PCR. We examined the proteins involved in the signaling pathways by Western blot analysis. We found that insulin and IGF1 enhanced IL-17-induced expression of Cxcl1, Ccl20, and Il-6, which was associated with increased phosphorylation of GSK3α and GSK3β in the presence of insulin and IGF1. AZD5363 inhibited the synergy between IL-17 and insulin/IGF1 through reducing phosphorylation of GSK3α and GSK3β by inhibiting Akt function. These findings imply that the cooperative crosstalk of IL-17 and insulin/IGF1 in initiating inflammatory responses may be alleviated by AZD5363.


Spectrum of resistance to root-knot nematodes and inheritance of heat-stable resistance in in pepper (Capsicum annuum L.).
Capsicum annuum L. has resistance to root-knot nematodes (RKN) (Meloidogyne spp.), severe polyphagous pests that occur world-wide. Several single dominant genes confer this resistance. Some are highly specific, whereas others are effective against a wide range of species. The spectrum of resistance to eight clonal RKN populations of the major Meloidogyne species, M. arenaria (2 populations), M. incognita (2 populations), M. javanica (1 population), and M. hapla (3 populations) was studied using eight lines of Capsicum annuum. Host susceptibility was determined by counting the egg masses (EM) on the roots. Plants were classified into resistant (R; EM ≤ 5) or susceptible (H; EM >5) classes. The french cultivar Doux Long des Landes was susceptible to all nematodes tested. The other seven pepper lines were highly resistant to M. arenaria, M. javanica and one population of M. hapla. Variability in resistance was observed for the other two populations of M. hapla. Only lines PM687, PM217, Criollo de Morelos 334 and Yolo NR were resistant to M. incognita. To investigate the genetic basis of resistance in the highly resistant line PM687, the resistance of two progenies was tested with the two populations of M. incognita: 118 doubled-haploid (DH) lines obtained by androgenesis from F(1) hybrids of the cross between PM687 and the susceptible cultivar Yolo Wonder, and 163 F(2) progenies. For both nematodes populations, the segregation patterns 69 R / 49 S for DH lines and 163 R / 45 S for F(2) progenies were obtained at 22°C and at high temperatures (32°C and 42°C). The presence of a single dominant gene that totally prevented multiplication of M. incognita was thus confirmed and its stability at high temperature was demonstrated. This study confirmed the value of C. annuum as a source of complete spectrum resistance to the major RKN.


Fear and loathing in the ER: managing procedural pain and anxiety in the Pediatric Emergency Department.
The pediatric emergency department can be frightening for children. Visits are unplanned, and frequently accompanied by significant emotional and physical distress. While treatment of pain and anxiety in children have been historically inadequate, the barriers to their treatment have largely been overcome through increased awareness, child- and family-focused care, standardized assessment, institutional safety protocols, and newer pharmacologic agents. The pediatric emergency physician is now a primary advocate for treatment of children's pain and anxiety and for the safe and appropriate use of procedural sedation. This article focuses on the treatment spectrum available for providing safe and effective procedural sedation, analgesia and anxiolytic therapy.


Biological Responses to Short-Term Maximal Exercise in Male Police Officers.
The specifics of short-term physical exercise are similar to the immediate reaction demands placed on police officers. Identifying the physiological predisposition to short-term high-intensity exercise in male law enforcement officers will assist in understanding their metabolism and make a significant contribution to a much more personal and individualized workout program. This will improve physical fitness of individual officers, improving their preparedness for such times of emergency. This cross-sectional study was conducted to investigate the responses of hematological (erythrocytes, hemoglobin, hematocrit, leucocytes, monocytes, neutrophils, lymphocytes), hormonal (testosterone, cortisol, melatonin), biochemical (glucose, uric-acid, lactate, creatine-phosphokinase) data to short-term maximal exercise in male police officers (n = 20). Blood samples were collected before- and after- the running-based anaerobic sprint test (RAST), and biological values were corrected for fluid shifts. Data were mean ± standard deviation of differences (= after minus before RAST). After the RAST, values of cortisol, lactate, neutrophils, lymphocytes, and monocytes increased significantly by 7.01 ± 37.36 mmol/l, 7.55 ± 1.67 mmol/l, 0.17 ± 0.26 103/µl, 0.61 ± 0.28 103/µl, and 0.10 ± 0.13 103/µl, respectively. After the RAST, values of melatonin, uric-acid, creatine-phosphokinase, hemoglobin, and hematocrit decreased significantly by -13.24 ± 4.60 pg/ml, -13.28 ± 14.35 µmol/l, -10.23 ± 10.13 IU/l, -2.01 ± 0.81 g/dl, and -4.46 ± 0.59%, respectively. Biological data of male police officers were affected by sprint test. Understanding changes in biological data following short-term maximal exercise can further assist in a better understanding of anaerobic metabolism, which will be helpful to find available methods for coaches to quantify training loads.


Molecular diversity of vascular potassium channel isoforms.
1. One essential role for potassium channels in vascular smooth muscle is to buffer cell excitation and counteract vasoconstrictive influences. Several molecular mechanisms regulate potassium channel function. The interaction of these mechanisms may be one method for fine-tuning potassium channel activity in response to various physiological and pathological challenges. 2. The most prevalent K+ channels in vascular smooth muscle are large-conductance calcium- and voltage-sensitive channels (maxi-K channels) and voltage-gated channels (Kv channels). Both channel types are complex molecular structures consisting of a pore-forming alpha-subunit and an ancillary beta-subunit. The maxi-K and Kv channel alpha-subunits assemble as tetramers and have S4 transmembrane domains that represent the putative voltage sensor. While most vascular smooth muscle cells identified to date contain both maxi-K and Kv channels, the expression of individual alpha-subunit isoforms and beta-subunit association occurs in a tissue-specific manner, thereby providing functional specificity. 3. The maxi-K channel alpha-subunit derives its molecular diversity by alternative splicing of a single-gene transcript to yield multiple isoforms that differ in their sensitivity to intracellular Ca2+ and voltage, cell surface expression and post- translational modification. The ability of this channel to assemble as a homo- or heterotetramer allows for fine-tuning control to intracellular regulators. Another level of diversity for this channel is in its association with accessory beta-subunits. Multiple beta-subunits have been identified that can arise either from separate genes or alternative splicing of a beta-subunit gene. The maxi-K channel beta-subunits modulate the channel's Ca2+ and voltage sensitivity and kinetic and pharmacological properties. 4. The Kv channel alpha-subunit derives its diverse nature by the expression of several genes. Similar to the maxi-K channel, this channel has been shown to assemble as a homo- and heterotetramer, which can significantly change the Kv current phenotype in a given cell type. Association with a number of the ancillary beta-subunits affects Kv channel function in several ways. Beta-subunits can induce inactivating properties and act as chaperones, thereby regulating channel cell-surface expression and current kinetics.


[Effects of different tillage methods on phospholipid fatty acids and enzyme activities in calcareous cinnamon soil].
In order to study changes of physical and chemical characteristics and microbial activities in soil under different tillage methods, effects of four tillage methods, rotary tillage (RT), subsoil tillage (ST), conventional tillage (CT) with corn straw returned to soil, and rotary tillage with no corn straw returned to soil (CK), on phospholipid fatty acids (PLFA) characteristics and hydrolase enzymes activities in calcareous cinnamon soil were investigated. The results showed that soil hydrolase enzymes activities, nutrient contents, microbial diversity varied greatly with the different tillage methods. Returning corn straw to soil increased the kinds, amount of soil total PLFAs, bacteria PLFAs and actonomycetes PLFAs, while decreased the fungi PLFAs, indicating that fungi was more adaptable than bacteria to an infertile environment. ST and CT resulted in higher amounts of total PLFAs, which were 74.7% and 53.3% higher than that of CK, indicating they were more beneficial to the growth of plants. They could also improve soil physical and chemical properties, increase alk-phosphatase, protease and urease activities, which would provide a favorable soil condition for high and stable crop yields.


[Acupuncture combined with traction therapy for lumbar disc herniation: a systematic review].
To evaluate the efficacy and safety of acupuncture combined with traction therapy for lumbar disc herniation, providing the basis for future research strategies. Randomized control trials. (RCT) of acupuncture combined with traction therapy for lumber disc herniation at home and abroad from 2000 to 2013 were searched, analysis and evaluation of literature and strength of evidence were based on the principles and methods of Evidence-based Medicine. The total effective rate and curative rate were considered as primary outcome measures; pain improvement, quality of life, relapse rate and adverse effects were considered as secondary outcome measures. Seventeen RCTs were identified, Meta-analysis showed that (1) total effective rate and curative rate: acupuncture combined with traction therapy was better than single therapy (acupuncture or traction); (2) pain improvement: acupuncture combined with traction therapy was better than traction therapy; (3) relapse rate: current evidence could not support the conclusion that acupuncture combined with traction therapy was better than traction therapy. Acupuncture combined with traction therapy for lumbar disc herniation was effective. However, the included studies were with high risk of bias, important outcome measures such as quality of life, relapse rate and adverse effects were not found in most of the studies. Current evidence has not yet been able to fully reflect acupuncture combined with traction therapy for lumbar disc herniation is better than single therapy, so more RCTs of higher quality are needed to further confirm its efficacy and safety.


Innate Immunomodulation in Food Animals: Evidence for Trained Immunity?
Antimicrobial resistance (AMR) is a significant problem in health care, animal health, and food safety. To limit AMR, there is a need for alternatives to antibiotics to enhance disease resistance and support judicious antibiotic usage in animals and humans. Immunomodulation is a promising strategy to enhance disease resistance without antibiotics in food animals. One rapidly evolving field of immunomodulation is innate memory in which innate immune cells undergo epigenetic changes of chromatin remodeling and metabolic reprogramming upon a priming event that results in either enhanced or suppressed responsiveness to secondary stimuli (training or tolerance, respectively). Exposure to live agents such as bacille Calmette-Guerin (BCG) or microbe-derived products such as LPS or yeast cell wall ß-glucans can reprogram or "train" the innate immune system. Over the last decade, significant advancements increased our understanding of innate training in humans and rodent models, and strategies are being developed to specifically target or regulate innate memory. In veterinary species, the concept of enhancing the innate immune system is not new; however, there are few available studies which have purposefully investigated innate training as it has been defined in human literature. The development of targeted approaches to engage innate training in food animals, with the practical goal of enhancing the capacity to limit disease without the use of antibiotics, is an area which deserves attention. In this review, we provide an overview of innate immunomodulation and memory, and the mechanisms which regulate this long-term functional reprogramming in other animals (e.g., humans, rodents). We focus on studies describing innate training, or similar phenomenon (often referred to as heterologous or non-specific protection), in cattle, sheep, goats, swine, poultry, and fish species; and discuss the potential benefits and shortcomings of engaging innate training for enhancing disease resistance.


Influence of Stimulus Polarity on the Auditory Brainstem Response From Level-Specific Chirp.
No known studies have investigated the influence of stimulus polarity on the Auditory Brainstem Response (ABR) elicited from level-specific (LS) chirp. This study is important as it provides a better understanding of the stimulus polarity selection for ABR elicited from LS chirp stimulus. We explored the influence of stimulus polarity on the ABR from LS chirp compared to the ABR from click at 80 dBnHL in normal-hearing adults. Nineteen adults with normal hearing participated. The ABRs were acquired using click and LS chirp stimuli using three stimulus polarities (rarefaction, condensation, and alternating) at 80 dBnHL. The ABRs were tested only on the right ear at a stimulus rate of 33.33 Hz. The ABR test was stopped when the recording reached the residual noise level of 0.04 µV. The ABRs amplitudes, absolute latencies, inter-peak latencies (IPLs), and the recorded number of averages were statistically compared among ABRs at different stimulus polarities and stimuli combinations. Rarefaction polarity had the largest ABR amplitudes and SNRs compared with other stimulus polarities in both stimuli. There were marginal differences in the absolute latencies and IPLs among stimulus polarities. No significant difference in the number of averages required to reach the stopping criteria was found. Stimulus polarities have a significant influence on the ABR to LS chirp. Rarefaction polarity is recommended for clinical use because of its larger ABR peak I, III, and V amplitudes than those of the other stimulus polarities.


Diagnosis and treatment of atypical and anaplastic meningiomas: a review.
Atypical and anaplastic meningiomas are uncommon tumors with a poorer prognosis than benign meningiomas. We reviewed the current literature and attempted to integrate and summarize available information to determine a logical approach to these tumors. Both tumors are rare and are often integrated with benign meningiomas when treatments are evaluated. In addition, because there has not been one histopathological classification scheme for atypical and anaplastic meningiomas in the past, there are numerous inconsistencies in the literature. Malignant progression with accumulation of mutations in a benign meningioma can result in an atypical and/or anaplastic meningioma. Both tumors are difficult to manage and have high recurrence and poor survival rates. The extent of tumor resection and histological grade are the key determinants for recurrence. In addition, metastases are unusual, but they do occur. We also review the evidence available that has resulted in the current World Health Organization classification. Radiation therapy can be used as an adjunctive treatment after both total and subtotal resection. In addition, the role of stereotactic radiosurgery is increasing, along with a possible role for brachytherapy. There are no effective chemotherapeutic agents available. A treatment algorithm is suggested.


Knockdown of SPOCK1 Inhibits the Proliferation and Invasion in Colorectal Cancer Cells by Suppressing the PI3K/Akt Pathway.
Sparc/osteonectin, cwcv, and kazal-like domains proteoglycan (testican) 1 (SPOCK1), known as testican-1, were found to be involved in the development and progression of tumors. However, in colorectal cancer (CRC), the expression pattern of SPOCK1 and its functional role remain poorly investigated. In the present study, we explored the role of SPOCK1 in CRC. Our results demonstrated that SPOCK1 is overexpressed in CRC cell lines. SPOCK1 silencing significantly inhibited the proliferation in vitro and the tumor growth in vivo. Furthermore, SPOCK1 silencing significantly attenuated the migration/invasion by reversing the EMT process in CRC cells. Finally, knockdown of SPOCK1 obviously decreased the protein expression levels of p-PI3K and p-Akt in HCT116 cells. In total, our study demonstrated for the first time that knockdown of SPOCK1 inhibits the proliferation and invasion in CRC cells, possibly through the PI3K/Akt signaling pathway. Therefore, SPOCK1 may be a potential therapeutic target for the treatment of CRC.


[Guideline of nitrous oxide-oxygen inhalation sedation technique in dental outpatient].
The use of nitrous oxide-oxygen inhalation sedation to relieve anxiety and pain in dental outpatient treatment has been a very mature and safe technique in the world. This technology has been introduced into China for nearly 20 years, and many clinical cases have proved its safety, practicability and effectiveness, which can meet the clinical needs of patients. To further standardize and popularize this technique, the Society of Sedation and Analgesia, Chinese Stomatological Association formed the recommended application guidelines after many discussions and revisions on the basis of widely soliciting opinions and referring to relevant literatures. It covers indications, contraindications, standardized operation procedures, occupational protection, identification and treatment of adverse reactions, training and other aspects. This guideline can be used as a reference for the use of nitrous oxide-oxygen inhalation sedation techniques in the outpatient setting of dentistry.


Effect of respiration on venous return and stroke volume in cardiac tamponade.
Measurements have been made of pressure and blood velocity in venae cavae and aorta in a patient with severe cardiac tamponade in whom there was pulsus paradoxus. The characteristic pressure changes of pulsus paradoxus were associated with variations in peak blood velocity in the ascending aorta and stroke output. Maximum filling of the right side of the heart occurred during inspiration, and was associated in time with minimum left ventricular stroke volume. These findings point to competition for space by the ventricles in the distended pericardial sac as being the major factor in the production of pulsus paradoxus.


The outcomes of mental health training programs: a review of the literature.
In these times of fiscal austerity, educators are asked increasingly to justify their programs. This review of literature indicates there are few controlled studies focusing on outcomes of mental health training programs, largely because the goals of training programs have been poorly identified and/or infrequently used as a basis for evaluation. In addition, these studies lack adequate criteria of success and standardized measures of success. Educators are, therefore, unable to critically evaluate the impact of the trained worker on patient care.


Effect of lifetime cigarette consumption on time domain spirogram indices.
A cross-sectional analysis on the dose-related change in time domain spirogram indices induced by lifetime cigarette consumption was conducted to examine the ability of those indices to detect early changes in the lung periphery in comparison with conventional spirometric indices. The subjects were asymptomatic healthy male workers from three occupational cohorts including asbestos workers and welders. They were asked to perform the forced expiration maneuver at least three times to obtain reliable results. A total of 893 subjects were enrolled in the study, and 484 of them who were aged 30 years or more and were free from chronic respiratory symptoms and abnormalities in chest radiography and spirometry, were analyzed. Although conventional indices, such as forced expiratory volume in one second and maximal midexpiratory flow, were not significantly different between smokers and nonsmokers, the standard deviation of transit times and of time constant distribution in smokers were significantly elevated compared with nonsmokers. Furthermore a dose-related change according to lifetime cigarette consumption was observed in those indices. We conclude that time domain spirogram indices, especially the standard deviation of time constant distribution, would be more useful than conventional indices in detecting early changes in the lung periphery.


Molecular Imaging of Tumors Using a Quantitative T 1 Mapping Technique via Magnetic Resonance Imaging.
Magnetic resonance imaging (MRI) of glioblastoma multiforme (GBM) with molecular imaging agents would allow for the specific localization of brain tumors. Prior studies using T 1-weighted MR imaging demonstrated that the SBK2-Tris-(Gd-DOTA)3 molecular imaging agent labeled heterotopic xenograft models of brain tumors more intensely than non-specific contrast agents using conventional T 1-weighted imaging techniques. In this study, we used a dynamic quantitative T 1 mapping strategy to more objectively compare intra-tumoral retention of the SBK2-Tris-(Gd-DOTA)3 agent over time in comparison to non-targeted control agents. Our results demonstrate that the targeted SBK2-Tris-(Gd-DOTA)3 agent, a scrambled-Tris-(Gd-DOTA)3 control agent, and the non-specific clinical contrast agent Optimark(™) all enhanced flank tumors of human glioma cells with similar maximal changes on T 1 mapping. However, the retention of the agents differs. The non-specific agents show significant recovery within 20 min by an increase in T 1 while the specific agent SBK2-Tris-(Gd-DOTA)3 is retained in the tumors and shows little recovery over 60 min. The retention effect is demonstrated by percent change in T 1 values and slope calculations as well as by calculations of gadolinium concentration in tumor compared to muscle. Quantitative T 1 mapping demonstrates the superior binding and retention in tumors of the SBK2-Tris-(Gd-DOTA)3 agent over time compared to the non-specific contrast agent currently in clinical use.


Lithographically Encrypted Inverse Opals for Anti-Counterfeiting Applications.
Colloidal photonic crystals possess inimitable optical properties of iridescent structural colors and unique spectral shape, which render them useful for security materials. This work reports a novel method to encrypt graphical and spectral codes in polymeric inverse opals to provide advanced security. To accomplish this, this study prepares lithographically featured micropatterns on the top surface of hydrophobic inverse opals, which serve as shadow masks against the surface modification of air cavities to achieve hydrophilicity. The resultant inverse opals allow rapid infiltration of aqueous solution into the hydrophilic cavities while retaining air in the hydrophobic cavities. Therefore, the structural color of inverse opals is regioselectively red-shifted, disclosing the encrypted graphical codes. The decoded inverse opals also deliver unique reflectance spectral codes originated from two distinct regions. The combinatorial code composed of graphical and optical codes is revealed only when the aqueous solution agreed in advance is used for decoding. In addition, the encrypted inverse opals are chemically stable, providing invariant codes with high reproducibility. In addition, high mechanical stability enables the transfer of the films onto any surfaces. This novel encryption technology will provide a new opportunity in a wide range of security applications.


Viral Myocarditis in the Setting of Delayed Manifestation of Hamman-Rich Syndrome.
Acute respiratory distress syndrome (ARDS)-like symptoms and rapid progression characterize the interstitial lung disease known as acute interstitial pneumonitis, also known as Hamman-Rich syndrome. It has a bad prognosis and a high incidence of mortality. We describe the case of a 25-year-old male patient with acute-onset type I respiratory failure with detrimental X-ray abnormalities who presented to the emergency room without any history of pulmonary disease or smoking. The provisional diagnosis of Hamman-Rich syndrome was reached after other clinical entities were ruled out based on CT findings. Myocardial hypokinesis of the apex and septum, as well as a modest systolic dysfunction (ejection fraction: 50%) similar to acute myocarditis, were detected by echocardiogram. Acute myocarditis in the setting of Hamman-Rich syndrome has been anecdotally reported and its mechanism remains to be elucidated.


Retrospective Comparative Analysis of Opioid Use and Intra-Articular Corticosteroid Injections Before and After Initial Treatment of Knee Osteoarthritis with Hylan G-F 20.
Knee osteoarthritis (OA) is a painful condition affecting >250 million people worldwide and is a leading cause of disability. Intra-articular (IA) corticosteroids and/or oral opioids are often recommended for the management of knee OA pain. There are, however, concerns regarding their safety and tolerability. Do patients diagnosed with knee OA show a decrease in opioids or IA corticosteroid injections prescribed/administered in hospitals following hylan G-F 20 treatment? This case-crossover, retrospective study using Health Facts®, a de-identified electronic health records database, enrolled patients ≥18 years with knee OA treated with hylan G-F 20 between January 1, 2000 and March 31, 2016, with data within 6 months before/after treatment. Primary endpoints compared days on opioids, amounts of opioids, and number of IA corticosteroid injections before/after hylan G-F 20 treatment via paired t-tests. A total of 513 patients were qualified for analysis. In the opioid cohort, the average total number of days on opioids (N = 50; 5.0 vs 13.5 days; P = 0.007) and average total amount of opioids (N = 44; 165.4 morphine mg equivalents [MME] vs 493.7 MME; P = 0.013) were lower 6 months after hylan G-F 20 treatment than 6 months before treatment. In the IA corticosteroid cohort, the average number of IA corticosteroid injections decreased after hylan G-F 20 treatment (N = 36; 0.56 in the 6-month follow-up vs 1.39 before treatment; P < 0.0001). Additional time frames of 1-5 months before and after treatment were examined; similar conclusions were drawn for patients with >2 months of data. Patients with knee OA previously treated with opioids or IA corticosteroid injections who received hylan G-F 20 demonstrated statistically significant decreases in these medications >2 months following hylan G-F 20 treatment versus >2 months before treatment.


In silico activity profiling reveals the mechanism of action of antimalarials discovered in a high-throughput screen.
The growing resistance to current first-line antimalarial drugs represents a major health challenge. To facilitate the discovery of new antimalarials, we have implemented an efficient and robust high-throughput cell-based screen (1,536-well format) based on proliferation of Plasmodium falciparum (Pf) in erythrocytes. From a screen of approximately 1.7 million compounds, we identified a diverse collection of approximately 6,000 small molecules comprised of >530 distinct scaffolds, all of which show potent antimalarial activity (<1.25 microM). Most known antimalarials were identified in this screen, thus validating our approach. In addition, we identified many novel chemical scaffolds, which likely act through both known and novel pathways. We further show that in some cases the mechanism of action of these antimalarials can be determined by in silico compound activity profiling. This method uses large datasets from unrelated cellular and biochemical screens and the guilt-by-association principle to predict which cellular pathway and/or protein target is being inhibited by select compounds. In addition, the screening method has the potential to provide the malaria community with many new starting points for the development of biological probes and drugs with novel antiparasitic activities.


Efficient Arylation of 2,7-Naphthyridin-1(2H)-one with Diaryliodonium Salts and Discovery of a New Selective MET/AXL Kinase Inhibitor.
New 8-chloro-2-phenyl-2,7-naphthyridin-1(2H)-one building blocks bearing diverse substitutes on the 2-phenyl group were synthesized via an efficient diaryliodonium salt-based N-arylation strategy with the advantage of mild conditions, short reaction times, and high yields. A small combinatorial library of 8-amino substituted 2-phenyl-2,7-naphthyridin-1(2H)-one was further conveniently constructed based on the above chlorinated naphthyridinones and substituted aniline. Preliminary biochemical screening resulted in the discovery of the new 2,7-naphthyridone-based MET/AXL kinase inhibitors. More importantly, 17c (IC50,MET of 13.8 nM) or 17e (IC50,AXl of 17.2 nM) and 17i (IC50,AXl of 31.8 nM) can efficient selectively inhibit MET or AXL kinase, respectively, while commercial cabozantinib showed no selectivity. The further exploration of the 8-substituted 2-phenyl-2,7-naphthyridin-1(2H)-one combinatorial library would significantly accelerate the discovery of more potent and selective inhibitors against diverse kinases.


Covalent interaction and semiempirical modeling of small molecules.
The use of sophisticated methods for modeling complex systems is gaining ever more importance recently because they allow the design of material with properties tailored to specific applications. However, problems may arise from conflicts between different reaction pathways inherent in the wide variety of chemical elements used. This is reflected by the impossibility of exactly solving the Schroedinger equation or of exactly describing the exchange interaction in density functional theory when the system complexity increases. An alternative is offered by the application of semiempirical methods because they strongly reduce system complexity. Normally, this is accomplished by increasing the degree of the approximations to the detriment of the formalism generality. This work is aimed to improve the semiempirical electronegativity equalization method. This is accomplished by modifying the point charge Madelung potential with the introduction of covalent interaction to better describe the chemical bonds.


Quantum dynamics simulations of the 2D spectroscopy for exciton polaritons.
We develop an accurate and numerically efficient non-adiabatic path-integral approach to simulate the non-linear spectroscopy of exciton-polariton systems. This approach is based on the partial linearized density matrix approach to model the exciton dynamics with explicit propagation of the phonon bath environment, combined with a stochastic Lindblad dynamics approach to model the cavity loss dynamics. Through simulating both linear and polariton two-dimensional electronic spectra, we systematically investigate how light-matter coupling strength and cavity loss rate influence the optical response signal. Our results confirm the polaron decoupling effect, which is the reduced exciton-phonon coupling among polariton states due to the strong light-matter interactions. We further demonstrate that the polariton coherence time can be significantly prolonged compared to the electronic coherence outside the cavity.


Are Th17 cells an appropriate new target in the treatment of rheumatoid arthritis?
Th17 cells play crucial roles not only in host defense but also in many human autoimmune diseases and corresponding animal models. Although many of the fundamental principles regarding Th17 biology have been rapidly elucidated in mice, there remain numerous controversies regarding the differentiation, plasticity, and pathogenicity of human Th17 cells. In this review, we consider these open questions in comparison to what has already been clarified in mice, and discuss the potential impact of discoveries related to the Th17 pathway on the development of new therapeutic strategies in Th17 driven autoimmune diseases, specifically rheumatoid arthritis.


The expression on mouse lymphoid cells of Th-B, an antigen common to mouse B cells and thymus cells.
A new cell-surface antigen of mouse lymphocytes, which was found earlier on normal and malignant plasma cells and about half the normal thymus cell population, has been shown to be on mature B cells and on the least immunocompetent subpopulation of cells of the thymus. The antigen was first detected by the cytotoxic effect on relevant cells of an in vivo purified rabbit antiserum raised against cells of the mouse IgM-producing plasma cell tumor MOPC-104E. We have now used a cell-transfer method to examine the effect of this antiserum (RantiM104E) on immunocompetent lymphocytes of spleen, including separated splenic B and T cells and thymus and bone marrow cells. We have found the antigen on the immunocompetent B lymphocytes of the spleen, but not on any other of the immunocompetent cells examined. The method involves determining the effect of the antiserum on the development of plaque-forming cells in the spleens of irradiated mice that have received lymphocytes from immune or non-immune donors by treating the lymphocyyes with RantiM104E antiserum before transfer. Plaque-forming cells are suppressed when antiserum-treated spleen cells are transferred to the irradiated mice.This suppression was found to be due to action on only the splenic B cells. The helper activity of splenic T cells was not reduced significantly by antiserum treatment. Neither was the helper activity of thymus cells similarly treated reduced by the antiserum. We found, in fact, that the subpopulation of the thymus cells which is resistant to the antiserum was more immunocompetent than the total thymus cell population. In addition, the hydrocortisone-resistant thymus cells were also found to be resistant to the cytotoxic action of the antiserum. The antiserum had no effect on bone marrow cells in the cell-transfer procedure. The antigen involved, which we are designating "Th-B" appears to be a B cell line marker which appears relatively early during the differentiation of mouse B cells from precursor cells and is lost during maturation of T cells.


Delivering advanced therapies: the big pharma approach.
After two decades of focused development and some recent clinical successes, cell and gene therapy (CGT) is emerging as a promising approach to personalized medicines. Genetically engineered cells as a medical modality are poised to stand alongside or in combination with small molecule and biopharmaceutical approaches to bring new therapies to patients globally. Big pharma can have a vital role in industrializing CGT by focusing on diseases with high unmet medical need and compelling genetic evidence. Pharma should invest in manufacturing and supply chain solutions that deliver reproducible, high-quality therapies at a commercially viable cost. Owing to the fast pace of innovation in this field proactive engagement with regulators is critical. It is also vital to understand the needs of patients all along the patient care pathway and to establish product pricing that is accepted by prescribers, payers and patients.


A Vondrak low pass filter for IMU sensor initial alignment on a disturbed base.
The initial alignment of the Inertial Measurement Unit (IMU) is an important process of INS to determine the coordinate transformation matrix which is used in the integration of Global Positioning Systems (GPS) with Inertial Navigation Systems (INS). In this paper a novel alignment method for a disturbed base, such as a vehicle disturbed by wind outdoors, implemented with the aid of a Vondrak low pass filter, is proposed. The basic principle of initial alignment including coarse alignment and fine alignment is introduced first. The spectral analysis is processed to compare the differences between the characteristic error of INS force observation on a stationary base and on disturbed bases. In order to reduce the high frequency noise in the force observation more accurately and more easily, a Vondrak low pass filter is constructed based on the spectral analysis result. The genetic algorithms method is introduced to choose the smoothing factor in the Vondrak filter and the corresponding objective condition is built. The architecture of the proposed alignment method with the Vondrak low pass filter is shown. Furthermore, simulated experiments and actual experiments were performed to validate the new algorithm. The results indicate that, compared with the conventional alignment method, the Vondrak filter could eliminate the high frequency noise in the force observation and the proposed alignment method could improve the attitude accuracy. At the same time, only one parameter needs to be set, which makes the proposed method easier to implement than other low-pass filter methods.


Effect of NaCl and mannitol iso-osmotic stresses on proline and free polyamine levels in embryogenic Fraxinus angustifolia callus.
With the aim to differentiate the ionic and osmotic components of salt stress, short and long-term changes in free polyamines and proline induced by iso-osmotic concentrations of NaCl (0.1 mol/L and 0.2 mol/L) and mannitol (0.2 mol/L and 0.4 mol/L) were determined in Fraxinus angustifolia callus. The peculiarities of the short-term responses were: i) a very early (30 min) and temporary increase in Putrescine (Pu) and Spermine (Spm) as a consequence of salt treatment, and ii) a continuous accumulation of Spermidine (Spd) and Spm in response to mannitol. The changes of Proline (Pro) were quite limited both in the short and in the long term, and generally occurred later than Polyamine (PAs) changes took place, suggesting a regulatory mechanism of PAs metabolism on Pro biosynthesis. In the long-term, no drastic accumulations of Pro or PAs in response to NaCl and mannitol were observed, suggesting that their physiological role is unlikely to be that of osmo-compatible solutes in this plant system. The salt induced a higher callus growth inhibition effect than did mannitol and this inhibition was associated with the reduction of endogenous levels of PAs, especially Pu. However, while a diverging time course was observed under lethal salt concentration (0.2 mol/L NaCl), a high parallelism in the endogenous changes of Pro and Pu was observed under all non-lethal conditions (control--0.2 and 0.4 mol/L mannitol--0.1 mol/L NaCl). Therefore the synchronous changes of Pro and Pu can be considered as a physiological trait associated with cell survival. These results indicate a strong metabolic co-ordination between PAs and Pro pathways and suggest that the metabolic fluxes through these pathways start competing only when the stress level is high enough to be lethal for cells.


Cytotoxic proteins and therapeutic targets in severe cutaneous adverse reactions.
Severe cutaneous adverse reactions (SCARs), such as Stevens-Johnson syndrome (SJS) and toxic epidermal necrosis (TEN), are rare but life-threatening conditions induced mainly by a variety of drugs. Until now, an effective treatment for SJS/TEN still remains unavailable. Current studies have suggested that the pathobiology of drug-mediated SJS and TEN involves major histocompatibility class (MHC) I-restricted activation of cytotoxic T lymphocytes (CTLs) response. This CTLs response requires several cytotoxic signals or mediators, including granulysin, perforin/granzyme B, and Fas/Fas ligand, to trigger extensive keratinocyte death. In this article, we will discuss the cytotoxic mechanisms of severe cutaneous adverse reactions and their potential applications on therapeutics for this disease.


Synergistic effects of cinnamaldehyde in combination with eugenol against wood decay fungi.
The combined effects of using cinnamaldehyde with catechin, quercetin or eugenol against wood decay fungi were examined by comparing their isoeffective concentrations to that of individual compound. Among all combinations, cinnamaldehyde with eugenol revealed the strongest synergy against Laetiporus sulphureus. The synergism was due to the interference of fungal cell wall synthesis and cell wall destruction plus radical scavenging effect. Results also suggested that antioxidant with fungicidal effect might be a better candidate than pure antioxidant for the system of fungicide/antioxidant.


The relationship between success of free gingival grafts and transplant thickness. Revascularization and shrinkage--a one year clinical study.
The purpose of this clinical investigation was to determine what effects the thickness of a free gingival graft has on the processes of revascularization and shrinkage. With the aid of a Mucotome, very thin, thin, and intermediate thickness grafts were excised from the palate. These were placed in the mandible or maxilla of patients with insufficient attached gingiva. Vertical graft shrinkage was measured at 15, 30, 90, and 360 days after surgery. Angiographic studies were made on 24 healing grafts. From the day of grafting to 360 days afterward the percentages of shrinkage were: very thin, 45%; thin, 44%; intermediate, 38%. The angiographic study demonstrated that rapid revascularization can be expected when uniform grafts of thin to intermediate thickness are placed on a periosteal recipient site which has been carefully freed of loose connective tissue and muscle attachments. An uneven, thick graft placed on a site of denuded bone favored a prolonged period of revascularization and delayed healing.


Surface-enhanced Raman gene probe for HIV detection.
We report, for the first time, the use of surface-enhanced Raman (SERS)-active labels for primers used in polymerase chain reaction amplification of specific target DNA sequences. This method has the potential for combining the spectral selectivity and high sensitivity of the SERS technique with the inherent molecular specificity offered by DNA sequence hybridization. The effectiveness of the detection scheme is demonstrated using the gag gene sequence of the human immunodeficiency virus. The potential use of multiple probes for simultaneous detection of multiple biological targets is discussed.


An obesity paradox in acute heart failure: analysis of body mass index and inhospital mortality for 108,927 patients in the Acute Decompensated Heart Failure National Registry.
Prior studies on chronic systolic heart failure (HF) have demonstrated that body mass index (BMI) is inversely associated with mortality, the so-called obesity paradox. The aim of this study was to determine whether BMI influences the mortality risk in acute decompensated HF, a subject not previously studied. The Acute Decompensated Heart Failure National Registry was analyzed for acute HF hospitalizations in 263 hospitals in the United States from October 2001 through December 2004. Patients with documented height and weight were divided into BMI (measured in kilograms per square meter) quartiles. Inhospital mortality by BMI quartile for all the patients and for those with reduced (n = 43,255) and preserved (n = 37,901) systolic function was assessed. Body mass index quartiles in the 108,927 hospitalizations were QI (16.0-23.6 kg/m2), QII (23.7-27.7 kg/m2), QIII (27.8-33.3 kg/m2), and QIV (33.4-60.0 kg/m2). Patients in the higher BMI quartiles were younger, had more diabetes, and had a higher left ventricular ejection fraction. Inhospital mortality rates decreased in a near-linear fashion across successively higher BMI quartiles. After adjustments for age, sex, blood urea nitrogen, blood pressure, creatinine, sodium, heart rate, and dyspnea at rest, BMI quartile still predicted mortality risk. For every 5-U increase in BMI, the odds of risk-adjusted mortality was 10% lower (95% CI 0.88-0.93, P < .0001). In this cohort of hospitalized patients with HF, higher BMI was associated with lower inhospital mortality risk. The relationship between BMI and adverse outcomes in HF appears to be complex and deserving of further study.


Prognostic role of miR-9 expression in various human malignant neoplasms: a meta-analysis.
Emerging evidence has shown that aberrant microRNA expression has the potential to be used for predicting survival and treatment response of malignant neoplasms. In recent years, the role of miR-9 had been investigated in various types of cancers, and it was found that the results were inconsistent and inconclusive. Hence, in this study, a meta-analysis was conducted to assess the prognostic value of miR-9 in various types of tumors. Eligible studies were identified through a systematic search in PubMed and EMBASE and then were assessed by further quality evaluation. Pooled hazard ratios (HRs) with 95% confidence intervals for overall survival (OS) were calculated to investigate the association between miR-9 expression and cancer prognosis. The pooled results of eight published studies showed that elevated miR-9 was a predictor of poor survival of various carcinomas, with pooled HR of 3.04 (95% confidence interval: 1.96-4.73) for OS. Subgroup analysis on the basis of tumor type, sample size, and HR estimate also showed that high levels of miR-9 were also significantly correlated with OS. In addition, when the subgroup analyses were grouped by follow-up time, it was found that the elevated expression of miR-9 was associated with a lower long-term survival when the follow-up time was >60 months, but there was no correlation between the outcomes and those patients whose follow-up time was <60 months. Funnel plots and Egger's tests revealed that there was no obvious publication bias risk in the meta-analysis. In conclusion, our results demonstrated that higher expression level of miR-9 significantly predicted worse OS in various carcinomas and that miR-9 may act as a novel biomarker in the prognosis of malignant neoplasms.


New rapid latex agglutination test for diagnosing Trichomonas vaginalis infection.
A newly developed latex agglutination test for Trichomonas vaginalis infection was compared for sensitivity, specificity, efficiency, and positive and negative predictive values with microscopy, culture, and an enzyme linked immunosorbent assay (ELISA) in the diagnosis of 395 women attending a genitourinary medicine clinic. T vaginalis infection was diagnosed in 42 (11%) women. The sensitivities of both the latex agglutination test and the ELISA were 95% compared with 74% for microscopy and 76% for culture. The latex test was specific and showed no cross reaction with a wide range of other genital tract infections. The latex agglutination test can detect antigen in both soluble and insoluble forms, and as it is simple to perform, can be undertaken during routine examination without recourse to special equipment or training. Further evaluation is required.


Structural insights on TRPV5 gating by endogenous modulators.
TRPV5 is a transient receptor potential channel involved in calcium reabsorption. Here we investigate the interaction of two endogenous modulators with TRPV5. Both phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and calmodulin (CaM) have been shown to directly bind to TRPV5 and activate or inactivate the channel, respectively. Using cryo-electron microscopy (cryo-EM), we determined TRPV5 structures in the presence of dioctanoyl PI(4,5)P2 and CaM. The PI(4,5)P2 structure reveals a binding site between the N-linker, S4-S5 linker and S6 helix of TRPV5. These interactions with PI(4,5)P2 induce conformational rearrangements in the lower gate, opening the channel. The CaM structure reveals two TRPV5 C-terminal peptides anchoring a single CaM molecule and that calcium inhibition is mediated through a cation-π interaction between Lys116 on the C-lobe of calcium-activated CaM and Trp583 at the intracellular gate of TRPV5. Overall, this investigation provides insight into the endogenous modulation of TRPV5, which has the potential to guide drug discovery.


Insights on the structure-activity relationship of peptides derived from Sticholysin II.
Sticholysin II (StII) is a pore-forming actinoporin from the sea anemone Stichodactyla helianthus. A mechanistic model of its action has been proposed: proteins bind to cell membrane, insert their N-termini into the lipid core and assemble into homo-tetramer pores responsible for host-cell death. Because very likely the first 10 residues of StII N-terminus are critical for membrane penetration, to dissect the molecular details of that functionality, we studied two synthetic peptides: StII1-30 and StII16-35 . They show diverse haemolytic and candidacidal activity that correlate with distinct orientations in SDS micelles. NMR shows that StII1-30 partly inserts into the micelle, while StII16-35 lays on the micelle surface. These results justify the diverse concentration dependence of their candidacidal activity supposing a different mechanism of action and providing new hints on StII lytic activity at molecular level. Biotechnological application of these peptides, focused on the development of therapeutic immunocomplexes, may be envisaged.


Responses of absolute and specific enzyme activity to consecutive application of composted sewage sludge in a Fluventic Ustochrept.
Composted sewage sludge (CS) is considered a rich source of soil nutrients and significantly affects the physical, chemical, and biological characteristics of soil, but its effect on specific enzyme activity in soil is disregarded. The present experiment examined the absolute and specific enzyme activity of the enzymes involved in carbon, nitrogen, and phosphorus cycles, the diversity of soil microbial functions, and soil community composition in a Fluventic Ustochrept under a maize-wheat rotation system in North China during 2012-2015. Application of CS led to increase in MBC and in its ratio to both total organic carbon (TOC) and microbial biomass nitrogen (MBN). Absolute enzyme activity, except that of phosphatase, increased in CS-treated soils, whereas specific activity of all the enzymes declined, especially at the highest dose of CS (45 t ha-1). The diversity of soil microbial community also increased in CS-treated soils, whereas its functional diversity declined at higher doses of CS owing to the lowered specific enzyme activity. These changes indicate that CS application induced the domination of microorganisms that are not metabolically active and those that use resources more efficiently, namely fungi. Redundancy analysis showed that fundamental alterations in soil enzyme activity depend on soil pH. Soil specific enzyme activity is affected more than absolute enzyme activity by changes in soil properties, especially soil microbial activity and composition of soil microflora (as judged by the following ratios: MBC/TOC, MBC/MBN, and TOC/LOC, that is labile organic carbon) through the Pearson Correlation Coefficient. Specific enzyme activity is thus a more accurate parameter than absolute enzyme activity for monitoring the effect of adding CS on the activities and structure of soil microbial community.


Time2bHealthy - An online childhood obesity prevention program for preschool-aged children: A randomised controlled trial protocol.
The use of parent-focused internet-based programs for the prevention and treatment of childhood obesity has shown promise but there is an acknowledged gap in parent-focused interventions which target the early childhood stage. The aim of this study is to determine the efficacy of Time2bHealthy - an online healthy lifestyle program for parents of preschool-aged children. The program will be evaluated using a two-arm, parallel, randomised controlled design. The 11-week program is underpinned by Social Cognitive Theory and consists of interactive modules on healthy eating, physical activity, screen-time and sleep. The intervention also involves elements of social media, where participants share discuss ideas and experiences and they can interact and obtain information with experts. Time2bHealthy is being compared to a comparison condition. Outcomes include change in BMI (primary outcome), dietary intake, physical activity, sleep, child feeding, parental role-modelling and parent self-efficacy. Process evaluation data, such as adherence and engagement with the online forums, will also be collected. Time2bHealthy is the first randomised controlled trial to our knowledge to assess the efficacy of an online parent-focussed healthy lifestyle program for preschool-aged children in changing body mass index. Early childhood is a crucial time for establishing healthy lifestyle behaviours and parents play an important role. This study therefore fills an important gap in the literature. If found to be efficacious, Time2bHealthy has potential for broad-reach access and translation into primary health care services. The study is registered with the Australian and New Zealand Clinical Trials Registry (12616000119493).


Obscurant Segmentation in Long Wave Infrared Images Using GLCM Textures.
The benefits of autonomous image segmentation are readily apparent in many applications and garners interest from stakeholders in many fields. The wide range of benefits encompass applications ranging from medical diagnosis, where the shape of the grouped pixels increases diagnosis accuracy, to autonomous vehicles where the grouping of pixels defines roadways, traffic signs, other vehicles, etc. It even proves beneficial in many phases of machine learning, where the resulting segmentation can be used as inputs to the network or as labels for training. The majority of the available image segmentation algorithmic development and results focus on visible image modalities. Therefore, in this treatment, the authors present the results of a study designed to identify and improve current semantic methods for infrared scene segmentation. Specifically, the goal is to propose a novel approach to provide tile-based segmentation of occlusion clouds in Long Wave Infrared images. This work complements the collection of well-known semantic segmentation algorithms applicable to thermal images but requires a vast dataset to provide accurate performance. We document performance in applications where the distinction between dust cloud tiles and clear tiles enables conditional processing. Therefore, the authors propose a Gray Level Co-Occurrence Matrix (GLCM) based method for infrared image segmentation. The main idea of our approach is that GLCM features are extracted from local tiles in the image and used to train a binary classifier to provide indication of tile occlusions. Our method introduces a new texture analysis scheme that is more suitable for image segmentation than the solitary Gabor segmentation or Markov Random Field (MRF) scheme. Our experimental results show that our algorithm performs well in terms of accuracy and a better inter-region homogeneity than the pixel-based infrared image segmentation algorithms.


High-resolution separation of disaccharide and oligosaccharide alditols from chondroitin sulphate, dermatan sulphate and hyaluronan using CarboPac PA1 chromatography.
Recent literature indicates that specific glycosaminoglycan structures are involved in various biological processes, such as anticoagulation, growth factor activation and viral infection. The initial step in the structural analysis of glycosaminoglycans is a definitive compositional analysis of its characteristic disaccharide repeat structures. Current chromatographic or electrophoretic procedures may have limitations in analysing glycosaminoglycan samples that are in low abundance, contain novel structures that need to be further characterized, or are metabolically labelled from radioactive precursors as a result of biosynthetic experiments. This study presents a new methodology for analysing disaccharides and oligosaccharides derived from chondroitin sulphate, dermatan sulphate and hyaluronan that fulfils the above criteria. The procedure involves the separation of reduced forms of these glycoconjugates on a CarboPac PA1 column using alkaline eluants. This study adopted a strategy which uses specific enzymes to release these disaccharides from their glycosaminoglycan forms. A borohydride reduction reaction was modified to be compatible with the buffer conditions commonly used with these enzymes in order to quantitatively reduce the disaccharides to their alditol forms (thereby stabilizing them to alkaline pH). Chromatography conditions were established which separated all known disaccharide alditol structures from chondroitin sulphate, dermatan sulphate and hyaluronan with extremely high resolution in a single run. Integrated pulsed amperometry was compared to UV absorbance measurement at 232 nm as two sensitive methods for detecting these reduced disaccharides; most of them could be routinely detected in the range of 50-500 ng. Data are presented applying this method to quantify hyaluronan in a biological sample which contains approximately 5000 cells and only approximately 10 ng of hyaluronan. Additional data are presented to demonstrate that this procedure will also separate oligosaccharide alditols derived from hyaluronan.


Exoscope for Upper Extremity Peripheral Nerve Surgery: Revision Carpal Tunnel Release With Epineurolysis and Hypothenar Fat Flap.
The exoscope is a high-resolution three-dimensional external microscope that allows the surgeon to perform delicate dissection in multiple regions of the body. The exoscope was originally used for intracranial and spine surgery. In this article, we describe its novel use in upper extremity peripheral nerve decompression surgery after recurrent carpal tunnel syndrome. This surgery is typically performed under the microscope, which allows precise microsurgical dissection to distinguish scar tissue from healthy nerve fascicles. Our case report highlights a 70-year-old man with recurrent carpal tunnel syndrome who underwent revision carpal tunnel surgery with epineurolysis and hypothenar fat flap. The ergonomic benefits of using the exoscope for microsurgery are described, along with intraoperative photos. Adequate symptom resolution was achieved.


Study of pubertal development in Abruzzo (Italy) and analysis of factors implicated in puberty variability.
The aim of the study was to confirm whether there is a secular trend towards the early onset of puberty without an early onset of menarche. The study included a total of 1,266 subjects, 771 females and 495 males, with an age range between 5.9 and 18.2 years. In all subjects height, percentile of height, weight, percentage of overweight, bone age and pubertal stages were evaluated. On the basis of pubertal stage the population was subdivided into Tanner's corresponding classes. The obtained data for the female population showed a stabilized onset of puberty compared to the last decades and prolonged length of the pubertal period. Pubertal development in males, instead, did not appear to have changed over the last decades. The study is in line with American studies; the earlier onset of puberty verified in the last decades has slowed prolonging the length of the pubertal period. The specific mechanisms implicated in sexual maturation are not yet well known and need further studies.


[Prophecy and mental illness].
It is a well known platitude that a mentally ill person may "think that he is God" or "believes that he is the Messiah". Despite the generalization and shallowness of this attitude, sometimes psychotic patients indeed have delusions with contents of divine revelation, messianic assignments or prophetic power. In this current article we examine the different connections between prophecy and mental condition, especially psychotic. We present sources that combine prophecy and insanity, and also possible psychiatric interpretation of these situations. Finally, we present the attitude of the Rambam to prophecy and the personality characteristics of the prophet, limiting the possibility of the mentally ill patient who pretends to be a prophet.


Situational cue and strategy influence on speechreading.
The influence of situational cues and reasoning strategies on speechreading was studied in normal-hearing and minimally hearing-impaired adults. Everyday Speech Sentences (Davis & Silverman, 1968) produced by a female talker were videotaped without sound in the context of a visual background scene. A non-verbal acoustic signal was matched to each sentence and dubbed onto the recording. Thus, each test item consisted of an inaudible sentence presented with an optical and acoustic situational cue. The experimental task was structured so that subjects speechread sentences using deductive or inductive reasoning strategies. Results demonstrated greater success in speechreading when sentences were matched with related situational cues than when unrelated cues were provided. This difference was noted only when inductive reasoning was used. The findings suggest that situational cues are most useful to the speechreader when he can be assured that a strong relationship exists between the spoken message and the talker's situational background.


Availability and Readability of Online Patient Education Materials Regarding Regional Anesthesia Techniques for Perioperative Pain Management.
Patient education materials (PEM) should be written at a sixth-grade reading level or lower. We evaluated the availability and readability of online PEM related to regional anesthesia and compared the readability and content of online PEM produced by fellowship and nonfellowship institutions. With IRB exemption, we constructed a cohort of online regional anesthesia PEM by searching Websites from North American academic medical centers supporting a regional anesthesiology and acute pain medicine fellowships and used a standardized Internet search engine protocol to identify additional nonfellowship Websites with regional anesthesia PEM based on relevant keywords. Readability metrics were calculated from PEM using the TextStat 0.1.4 textual analysis package for Python 2.7 and compared between institutions with and without a fellowship program. The presence of specific descriptive PEM elements related to regional anesthesia was also compared between groups. PEM from 17 fellowship and 15 nonfellowship institutions were included in analyses. The mean (SD) Flesch-Kincaid Grade Level for PEM from the fellowship group was 13.8 (2.9) vs 10.8 (2.0) for the nonfellowship group (p = 0.002). We observed no other differences in readability metrics between fellowship and nonfellowship institutions. Fellowship-based PEM less commonly included descriptions of the following risks: local anesthetic systemic toxicity (p = 0.033) and injury due to an insensate extremity (p = 0.003). Available online PEM related to regional anesthesia are well above the recommended reading level. Further, fellowship-based PEM posted are at a higher reading level than PEM posted by nonfellowship institutions and are more likely to omit certain risk descriptions.


Pharmacologic approaches to butterfly wing patterning: sulfated polysaccharides mimic or antagonize cold shock and alter the interpretation of gradients of positional information.
Butterflies produce complex and diverse wing patterns by mechanisms that are generally unknown. We have employed a pharmacological approach to explore the molecular mechanisms of pattern formation. In a screen of over 200 compounds injected into developing Junonia coenia pupae, we identified several specific sulfated polysaccharides that caused widespread, dose-dependent effects on adult wing patterns. These compounds were well tolerated and permitted butterflies to eclose normally and take flight at moderate levels of effect. Heparin and closely related chondroitin sulfates caused stage-specific expansion of distal and proximal band systems and reduction and repatterning of eyespots. Dextran sulfate and fucoidan, whose structures are widely divergent from heparin and one another, caused contraction of distal and proximal systems, but had no effect on eyespots. Nonsulfated or nonpolymeric saccharides were without effect. Pattern alterations were indistinguishable from those reported for extreme cold shock and exposure to sodium tungstate and "molsin". When administered after cold shock or coinjected with heparin, dextran sulfate reversed all patterning effects. We suggest that the primary effect of polysaccharide treatments is to alter the interpretation of gradients of positional information along the proximodistal axis of the pupal wing.


Sex-dependent and ontogenetic effects of low dose ethanol on social behavioral deficits induced by mouse maternal separation.
Early life stress can induce lifelong emotional and social behavioral deficits that may in some cases be alleviated by drugs or alcohol. A model for early life stress, rodent maternal separation, recapitulates these behavioral sequelae, which are not limited to potentiated anxiety-like behavior, attenuated social motivation, and altered reward-seeking. Here we employed mouse maternal separation with early weaning (MSEW), consisting of pup-dam separation lasting 4-8 hours on postnatal days (PD) 2-16, with early weaning on PD 17. Prior MSEW studies have limited subjects by age or sex, so we more comprehensively investigated MSEW effects in both sexes, during adolescence and adulthood. We found universal effects of MSEW to include lifelong enhancement of anxiety-like and despair behavior, as well as deficits in social motivation. We also observed some sex-dependent effects of MSEW, namely that female MSEW mice exhibited social habituation to a greater degree than their male counterparts. Low dose ethanol administration had no major effects on the social behavior of non-stressed mice. But interestingly, MSEW-induced social habituation was counteracted by low dose ethanol in adolescent female mice, and potentiated in adolescent male mice. These effects were absent in adult animals, suggesting that ethanol may exert differential effects on the developing brain in such a manner to produce age-, sex-, and stress-dependent effects upon social behavior. Together, results indicate that MSEW reliably produces long-lasting impairments in emotional and social behaviors in both sexes and across the lifespan, but may exert more salient social behavioral effects on female animals.


Cross-resistance and Fitness Cost Analysis of Resistance to Thiamethoxam in Melon and Cotton Aphid (Hemiptera: Aphididae).
The melon/cotton aphid, Aphis gossypii Glover, is a notorious pest in many crops. The neonicotinoid insecticide thiamethoxam is widely used for A. gossypii control. To evaluate thiamethoxam resistance risk, a melon/cotton aphid strain with an extremely high level of resistance to thiamethoxam (>2,325.6-fold) was established after selection with thiamethoxam for 24 generations. Additionally, the cross-resistance pattern to other neonicotinoids and fitness were analyzed. The cross-resistance results showed the thiamethoxam-resistant strain had extremely high levels of cross-resistance against clothianidin (>311.7-fold) and nitenpyram (299.9-fold), high levels of cross-resistance against dinotefuran (142.3-fold) and acetamiprid (76.6-fold), and low cross-resistance against imidacloprid (9.3-fold). Compared with the life table of susceptible strain, the thiamethoxam-resistant strain had a relative fitness of 0.950, with significant decreases in oviposition days and fecundity and prolonged developmental duration. The molecular mechanism for fitness costs was studied by comparing the mRNA expression levels of juvenile hormone acid O-methyltransferase (JHAMT), juvenile hormone-binding protein (JHBP), juvenile hormone epoxide hydrolase (JHEH), ecdysone receptor (EcR), ultraspiracle protein (USP), and Vitellogenin (Vg) in the susceptible and thiamethoxam-resistant strains. Significant overexpression of JHEH and JHBP and downregulation of EcR and Vg expression were found in the thiamethoxam-resistant strain. These results indicate that A. gossypii has the potential to develop extremely high resistance to thiamethoxam after continuous exposure, with a considerable fitness cost and cross-resistance to other neonicotinoids.


Liver inclusion improves outcomes of intestinal retransplantation in adults. [Corrected].
Retransplantation is a viable treatment option for patients with primary intestinal graft loss. We analyzed outcomes of retransplantations (n = 23) in adults from May 2000 to May 2010 at our center and compared patients who received liver-free retransplantations (n = 13) with those who received liver-inclusive retransplantations (n = 10). The overall survival rates at 1, 3, and 5 years in retransplantations were 90.9%, 67.1%, and 59.7% (patient) and 82.2%, 58.6%, and 51.3% (graft), respectively, which were similar to the rates in primary transplants. The patient survival rates at 1, 3, and 5 years were 91.7%, 55.6%, and 41.7%, respectively, in liver-free retransplantations, as compared to rates of 90.0%, 80.0%, and 80.0% in liver-inclusive retransplantations. The graft survival rates at 1, 3, and 5 years in liver-free retransplantations were 76.2%, 40.6%, and 27.1%, respectively, which were significantly worse than those in liver-inclusive retransplantations (P = 0.03). Within an average follow-up of 32.3 months, 8 of 13 (61.5%) of liver-free retransplantations underwent enterectomy because of severe acute cellular rejection (n = 3) or chronic rejection (n = 5). Six of 13 (46.2%) recipients succumbed to rejection-related complications. Compared to liver-free retransplantations, the rate and severity of acute rejection were markedly de creased in liver-inclusive retransplantations, and no chronic rejection was seen. Within an average follow-up of 44.5 months, two of 10 (20%) died due to graft-versus-host disease and infection, respectively. A liver-inclusive retransplantation offers a better long-term patient and graft survival, suggesting that including the liver as part of an intestinal graft should be considered in adult recipients when retransplantation is necessary.


Directed Self-Assembly of Triblock Copolymer on Chemical Patterns for Sub-10-nm Nanofabrication via Solvent Annealing.
Directed self-assembly (DSA) of block copolymers (BCPs) is a leading strategy to pattern at sublithographic resolution in the technology roadmap for semiconductors and is the only known solution to fabricate nanoimprint templates for the production of bit pattern media. While great progress has been made to implement block copolymer lithography with features in the range of 10-20 nm, patterning solutions below 10 nm are still not mature. Many BCP systems self-assemble at this length scale, but challenges remain in simultaneously tuning the interfacial energy atop the film to control the orientation of BCP domains, designing materials, templates, and processes for ultra-high-density DSA, and establishing a robust pattern transfer strategy. Among the various solutions to achieve domains that are perpendicular to the substrate, solvent annealing is advantageous because it is a versatile method that can be applied to a diversity of materials. Here we report a DSA process based on chemical contrast templates and solvent annealing to fabricate 8 nm features on a 16 nm pitch. To make this possible, a number of innovations were brought in concert with a common platform: (1) assembling the BCP in the phase-separated, solvated state, (2) identifying a larger process window for solvated triblock vs diblock BCPs as a function of solvent volume fraction, (3) employing templates for sub-10-nm BCP systems accessible by lithography, and (4) integrating a robust pattern transfer strategy by vapor infiltration of organometallic precursors for selective metal oxide synthesis to prepare an inorganic hard mask.


Revising the ADAS-cog for a more accurate assessment of cognitive impairment.
To examine whether it is appropriate to sum the cognitive part of the Alzheimer Disease Assessment Scale (ADAS-cog) items to assess cognitive impairment. This assumes items to have (1) equal measurement precision and (2) hierarchically ordered categories. Rasch analysis on the basis of pooled data from 3 Randomized Controlled Trials was used to examine these assumptions and to estimate each patient's level of impairment. Analyses were replicated in an independent sample. The original ADAS-cog scoring did not fit the Rasch Model and did not reliably distinguish between impairment levels. Patients with equal test scores had different impairment levels. Similarly, patients with different test scores could have the same impairment level. Revising the ADAS-cog by (1) weighting the items by their measurement precision and (2) collapsing nonhierarchical item categories resulted in good fit and a valid one to one correspondence between sum scores and estimated impairment levels. This revealed that equal differences in ADAS-cog scores did not reflect equal differences in impairment level along the test's score range. It is appropriate to summate the ADAS-cog items provided that the items are weighted and have their categories hierarchically ordered.


[Therapeutic communication in Nursing: essential instrument of care].
The objective of the study was to analyze the process of therapeutic communication as developed by nurses who work in hospital units according to Peplau's theory. It is a descriptive-exploratory research accomplished in the period of October and November of 2006, of the Municipal district of Fortaleza, CE, Brazil in october and november, 2006. The participants of the study were 14 patients and 12 nurses, through a a semi-structured interview. Data were organized in thematic and discussed with literature review. Results demonstrated that since admission to hospital discharge, there is communication and interaction with a interpersonal relationship. Several times, however, that communication does not occur as it was supposed to, because nurse in her work does not priorize the visit to patients in her daily activities. Patients have difficulties to distinguish nurses from other health care professionals, which difficults the approach and development of therapeutic communication.


Immobilization of lipase B within micron-sized poly-N-isopropylacrylamide hydrogel particles by solvent exchange.
The aim of the present work is the use of a water soluble enzyme in an organic solvent, still with a pronounced catalytic activity. Therefore, lipase B from Candida antarctica (CalB) is immobilized within micron-sized thermosensitive p-NIPAM hydrogel particles using a solvent exchange from polar to organic solvents. The absorbed amount of CalB is investigated at different immobilization temperatures. Confocal laser scanning microscopy (CLSM) shows that CalB is homogeneously distributed within the polymer network. An enhanced specific activity of CalB in n-hexane is achieved after immobilization within the p-NIPAM microgels. In order to get information on the supply of the substrate depending on the temperature, the activity is determined at different reaction temperatures. Additionally, the system is stable in the organic solvent, namely n-hexane, and shows a good reusability.


CDK5 and MAPT Gene Expression in Alzheimer's Disease Brain Samples.
Alzheimer's disease (AD) is a neurodegenerative disorder characterized by extracellular amyloid plaque and neurofibrillary tangles in the brain. Studies have shown that neurons are able to re-enter the cell cycle, but not enough to enable full replication. This leads to cell death and consequent neurodegeneration. This study aimed to characterize the expression of the MAPT gene and CDK5 (the gene involved in cell cycle regulation) in brain samples from patients with AD and controls. The real-time-PCR technique was used to characterize 150 samples from three areas of the brain (entorhinal cortex, auditory cortex, and hippocampus) of 26 AD patients and 24 healthy elderly subjects. When the brain samples were analyzed collectively, a decrease in CDK5 and MAPT gene expression was found in AD patients. When each groups' samples were separated by area of the brain and compared, significant differences were found in CDK5 expression in the hippocampus and the entorhinal cortex. In both cases, mRNA was lower in the AD group (p=0.0001); however, the same analysis using the MAPT gene revealed no significant statistical differences. No statistical differences were found when gene expression was compared between the different regions of the brain within each group. These results may contribute to a better understanding of the involvement of CDK5 and MAPT genes in AD in that they consider different areas of the brain that are affected differently based on disease progression. The main challenge is to establish an effective therapy for this debilitating disease in the future.


[Spontaneous spinal extradural hematoma. A rare complication during anticoagulant therapy].
A 42-year-old woman with metallic valve prosthesis in aortic position, in use of oral anticoagulant, presented a sudden syndrome of medullary compression, characterized by intense dorsal pain and partial paralysis of the inferior portion of the body, progressing to flaccid paraplegia of the arms and legs with level of sensibility in T-2. Large extradural spinal hematoma of the thoracic spine was diagnosed by magnetic resonance with medulla compression in the proximal portion. The patient was underwent to surgery (decompressive laminectomy), however with little motricity improvement and the level of sensibility. Due to respiratory insufficiency the patient died.


Behavior of polyethylene glycol molecules at an oscillating solid-liquid interface.
The behavior of polyethylene glycol (PEG) molecules at an oscillating solid-liquid interface was examined using a quartz crystal microbalance (QCM). The number-average molecular weights (Mn) of PEG molecules were systematically varied. This study revealed that the shift in the energy dissipation factor, ΔD, of the QCM against square root of the density-viscosity product of the PEG solution is linear and has an intercept. Moreover, the systematical analysis revealed that the ΔD slope rapidly decreases with Mn, and the ΔD intercept becomes constant above 1.1×10(4) g/mol. Those results indicated that the resonant length of PEG molecules moving with the oscillating plate at 9 MHz is 97.6 Å. We also found that the difference between the resonant lengths calculated from ΔD and series resonance-frequency shift, ΔF, is related to the density of the thin PEG layer formed on the gold electrode.


Angiographic and immunologic studies in acute necrotizing jejunoileitis.
Eighteen cases of laparotomy-proven acute necrotizing jejunoileitis were treated during the period 1992-1998. Routine laboratory investigations and bacteriological studies were non-contributory towards establishing an etiological basis for the diagnosis. Superior mesenteric artery digital subtraction angiography, carried out in five patients, demonstrated extensive neovascularization. IgG, IgM and IgA were normal in all eight children in whom it was measured; IgE levels were grossly elevated (mean 900 IU/ml; normal range 0-100 IU/ml). Four cases had evidence of hypersensitivity reactions (type I in one patient and type III in three patients) on histopathological examination of the resected small bowel. In addition, histopathological examination also showed the neovascularization observed on angiography. The evidence suggests an immunologically mediated insult resulting in intense vasculitis, which in turn causes ischemic damage to the bowel wall.


Androgen receptor content of normal and hyperplastic human prostate.
In an effort to determine whether human benign prostatic hyperplasia (BPH) is characterized by an increase in androgen receptor content, the levels of nuclear and cytosolic androgen receptors were quantitated in normal prostatic tissue obtained from five young men (mean age /+- SEM, 26 +/- 3 yr) and in hyperplastic (periurethral) and peripheral prostatic tissues obtained from nine older men (mean age, 62 +/- 2 yr). There was no significant difference between the cytosolic or nuclear androgen receptor content of the hyperplastic, peripheral, or normal prostatic tissue. Thus, in this study we were unable to identify an increase in androgen receptor content in BPH. These findings fail to support the hypothesis that increases in androgen receptor content are involved in the pathogenesis of human BPH.


Assessment of carotid plaque vulnerability using structural and geometrical determinants.
Because many acute cerebral ischemic events are caused by rupture of vulnerable carotid atheroma and subsequent thrombosis, the present study used both idealized and patient-specific carotid atheromatous plaque models to evaluate the effect of structural determinants on stress distributions within plaque. Using a finite element method, structural analysis was performed using models derived from in vivo high-resolution magnetic resonance imaging (MRI) of carotid atheroma in 40 non-consecutive patients (20 symptomatic, 20 asymptomatic). Plaque components were modeled as hyper-elastic materials. The effects of varying fibrous cap thickness, lipid core size and lumen curvature on plaque stress distributions were examined. Lumen curvature and fibrous cap thickness were found to be major determinants of plaque stress. The size of the lipid core did not alter plaque stress significantly when the fibrous cap was relatively thick. The correlation between plaque stress and lumen curvature was significant for both symptomatic (p=0.01; correlation coefficient: 0.689) and asymptomatic patients (p=0.01; correlation coefficient: 0.862). Lumen curvature in plaques of symptomatic patients was significantly larger than those of asymptomatic patients (1.50+/-1.0 mm(-1) vs 1.25+/-0.75 mm(-1); p=0.01). Specific plaque morphology (large lumen curvature and thin fibrous cap) is closely related to plaque vulnerability. Structural analysis using high-resolution MRI of carotid atheroma may help in detecting vulnerable atheromatous plaque and aid the risk stratification of patients with carotid disease.


Are there critical periods for brain growth in children born preterm?
Children born very preterm who attend mainstream schools have a high prevalence of minor motor, behavioural, and learning disorders. These appear to be associated with reduced postnatal growth, particularly of the head. It is unclear when this poor growth occurs and whether growth restriction during different periods has different effects on later function. To identify periods during early development, in children born preterm, when impaired head growth may influence minor motor and cognitive function. A geographically defined cohort of 194 infants born in Merseyside during 1980-81 and weighing less than 1500 g. Measurements of head circumference (occipitofrontal circumference (OFC)) were available at birth, hospital discharge, 4 years, and 15 years of age. Assessments of intelligence (intelligence quotient (IQ)) and minor motor impairment (test of motor impairment (TOMI)) were made at 8 years of age. Clinical, social, and demographic variables were obtained from the clinical record and maternal interviews. IQ correlated significantly with OFC at 4 and 15 years of age after correction for growth restriction at birth (intrauterine growth restriction (IUGR)) and social class. TOMI scores correlated significantly with OFC at all four times, but especially with OFC at discharge and with change in OFC between birth and discharge. They were not affected by correction for social class or IUGR. Although both IQ and minor motor impairments correlate strongly with each other at school age in very low birthweight children, the factors determining them and their timing of operation are different. Interventions designed to improve IQ in this population would need to reduce IUGR and improve later childhood growth. Those aimed to improve motor ability need to be targeted more at brain protection during the neonatal period.


Associations of dual use of tobacco cigarettes and e-cigarettes, sleep duration, physical activity and depressive symptoms among middle-aged and older Korean adults.
There is limited evidence of the association between dual tobacco-e-cigarette use and health-related variables in Korea. Thus, this study aimed to investigate the associations between types of cigarette smoking, sleep duration, physical activity and depressive symptoms among Korean adults. A cross-sectional study design using the 2019 Korean Community Health Survey. The study subjects consisted of 179,004 adults older than 40 years from a total of 229,099 individuals. Self-reported general characteristics, smoking history, sleep duration, physical activity and depressive symptoms were analysed. In multinomial logistic regression, dual users of tobacco cigarettes and e-cigarettes were more likely to have sleep duration of less than 7 h per day and to report both mild and moderate-to-severe depressive symptoms than non-smokers. Single use of either cigarettes or e-cigarettes increased the risk of short sleep duration and moderate-to-severe depressive symptoms.


Inhibition of Hageman factor activation.
A method for studying inhibitors of the contact stages of blood coagulation is described. A number of positively charged substances were shown to inhibit the contact stages. The inhibitory substances include spermine, cytochrome c, ribonuclease, and lysozyme. The inhibitory effect of these substances was neutralized by the addition of an activated plasma thromboplastin antecedent, factor XI, (PTA) fraction. Other positively charged substances including protamine, hexadimethrine, polylysine, polyornithine, methylene blue, and ortho-toluidine blue also inhibited the contact stages of coagulation, but the inhibitory effect on coagulation was not neutralized by the activated PTA fraction. Negatively charged substances such as heparin and insulin did not inhibit the contact stages of coagulation. Cytochrome c inhibited Celite adsorption of a partially purified Hageman factor fraction, and cytochrome, ribonuclease, spermine, and lysozome inhibited the adsorption of Hageman factor from PTA-deficient plasma. Very much smaller quantities of Celite completely adsorbed Hageman factor from the fraction rather than from whole plasma, which suggested the possibility that plasma contains an inhibitor or inhibitors of Hageman factor adsorption. Furthermore cytochrome c, spermine, ribonuclease, and lysozyme inhibited the coagulant activity of the following activators of the Hageman and PTA factors: Celite, kaolin, sodium stearate, ellagic acid, and skin. It is suggested that negatively charged sites on these activators are critical for adsorption and activation and that inhibition results from neutralization of the negatively charged sites by the adsorbed inhibtor. Tests with polylysine polymers indicate that inhibitory activity is directly related to molecular size over the molecular weight range of 4000 to 100,000.


Purification of the yeast mitochondrial methionyl-tRNA synthetase. Common and distinctive features of the cytoplasmic and mitochondrial isoenzymes.
Yeast-mitochondrial methionyl-tRNA synthetase was purified 1060-fold from mitochondrial matrix proteins of Saccharomyces cerevisiae using a four-step procedure based on affinity chromatography (heparin-Ultrogel, tRNA(Met)-Sepharose, Agarose-hexyl-AMP) to yield to a single polypeptide of high specific activity (1800 U/mg). Like the cytoplasmic methionyl-tRNA synthetase (Mr 85,000), the mitochondrial isoenzyme is a monomer, but of significantly smaller polypeptide size (Mr 65,000). In contrast, the corresponding enzyme of Escherichia coli is a dimer (Mr 152,000) made up of identical subunits. The measured affinity constants of the purified mitochondrial enzyme for methionine and tRNA(Met) are similar to those of the cytoplasmic isoenzyme. However, the two yeast enzymes exhibit clearly different patterns of aminoacylation of heterologous yeast and E. coli tRNA(Met). Furthermore, polyclonal antibodies raised against the two proteins did not show any cross-reactivity by inhibition of enzymatic activity and by the highly sensitive immunoblotting technique, indicating that the two enzymes share little, if any, common antigenic determinants. Taken together, our results further support the belief that the yeast mitochondrial and cytoplasmic methionyl-tRNA synthetases are different proteins coded for by two distinct nuclear genes. Like the yeast cytoplasmic aminoacyl-tRNA synthetases, the mitochondrial enzymes displayed affinity for immobilized heparin. This distinguishes them from the corresponding enzymes of E. coli. Such an unexpected property of the mitochondrial enzymes suggests that they have acquired during evolution a domain for binding to negatively charged cellular components.


An investigation onto Cd toxicity to freshwater microalga Chlorella sorokiniana in mixotrophy and photoautotrophy: A Bayesian approach.
Aquatic ecosystems are composed by a myriad of dissolved organic materials that can be assimilated by microalgae, while they can perform photosynthesis, this is refereed as mixotrophy. However, ecotoxicological tests usually consider only the photoautotrophic metabolism. This research investigated the ecotoxicological differences between photoautotrophy and mixotrophy in Chlorella sorokiniana exposed to cadmium (Cd). Chlorophyll a, photosynthetic efficiency (Fv/Fm), cell viability, biochemical composition and pH were used to monitor possible toxic effects at 72 h cultures. Glucose (1 g.L-1) was used as organic carbon source. To evaluate the probability of the photoautotrophic culture being more affected by Cd than the mixotrophic one, Bayesian statistical analysis was performed. The photoautotrophic cultures were more affected by Cd than the mixotrophic ones, with reduction of all evaluated parameters, except for protein concentration. However, in mixotrophic cultures, no changes in protein concentration and proteins:carbohydrates ratio were observed, and chlorophyll a, Fv/Fm and cell viability were only affected at the high Cd concentrations (range ln -11.5 to -9.4). However, both mixotrophy and photoautotrophy had the same probability of having the carbohydrates concentration affected by Cd. We conclude that the microalgae in mixotrophy were more resistant to the Cd than in photoautotrophy. In addition, we showed that under photoautotrophy Fv/Fm decreased linearly as Cd concentration increased, but in mixotrophy no effect was observed up to 10-5 molL-1 Cd, after which it decreased. We rationale that the reduced photosynthetic capacity under mixotrophy can end up reducing the release of oxygen gas, which can compromise the entire aquatic ecosystem.


Investigation into the Role of PI3K and JAK3 Kinase Inhibitors in Murine Models of Asthma.
Asthma is a clinical disorder commonly characterized by chronic eosinophilic inflammation, remodeling and hyper responsiveness of the airways. However, the kinases like Phosphoinositide 3 kinase (PI3K) and Janus kinase 3 (JAK3) are involved in mast cell proliferation, activation, recruitment, migration, and prolonged survival of inflammatory cells. The present study was designed to evaluate the in-vivo comparative effects of two kinase inhibitors on airway inflammation and airway remodeling in acute and chronic models of asthma. Mice were sensitized twice intra-peritoneally and then challenged by inhalation with ovalbumin (OVA). They developed an extensive inflammatory response, goblet cell hyperplasia, collagen deposition, airway smooth muscle thickening similar to pathologies observed in human asthma. The effects of PI3K inhibitor (30 mg/kg, p.o), JAK3 inhibitor (30 mg/kg, p.o) and Dexamethasone (0.3 mg/kg) on airway inflammation and remodeling in OVA sensitized/challenged BALB/c mice were evaluated. Twenty-four hours after the final antigen challenge, bronchoalveolar lavage (BAL) and histological examinations were carried out. It was observed that kinase inhibitors significantly reduced airway inflammation as evidenced by the decrease in pro inflammatory cytokines in BALF and lung homogenate and inflammatory cell count in sensitized mice after allergen challenge. Lung histological analysis showed increased infiltration of inflammatory cells, hyperplasia of goblet cells and the collagen deposition, which were significantly reduced with kinase inhibitor. In conclusion, our data suggest that PI3K and JAK3 inhibitors showed promising alternative therapeutic activity in asthma, which might significantly counteract the airway inflammation in patients with allergic asthma.


Diffuse lipomatosis in the leg after poliomyelitis.
Two elderly women with gradual enlargement of the leg following paralytic poliomyelitis are reported. From the clinical appearance, plain radiograms and angiographic studies the enlargement was thought to be caused by adipose tissue. On exploration the muscles within the calfs were found to be entirely replaced by adipose tissue. Histological examination revealed diffuse lipomatosis involving skeletal muscle, fasciae and in one case even periosteum and bone. Peripheral nerves exhibited fibrosis and myelin degeneration and vessel walls showed unusual angioma-like changes. Since it is known from the literature that peripheral nerve lesions of different kinds can be followed by adipose overgrowth we believe that there is a pathogenetic relationship between poliomyelitis and the diffuse lipomatosis in the present cases.


Second Generation Drug-Eluting Stent Implantation versus Coronary Artery Bypass Grafting in the Treatment of Young Patients with Left Main and/or Multivessel Coronary Disease.
Many studies have compared the outcomes of coronary artery bypass graft (CABG) and percutaneous coronary intervention (PCI) for complex coronary artery disease (CAD). However, no trials have focused on young patients (<45 years) with complex CAD. We conducted a retrospective evaluation to compare the outcomes of a second-generation drug-eluting stent (DES) and CABG in young patients with LM or three-vessel disease. In young patients with complex CAD who underwent PCI or CABG, a Kaplan-Meier analysis and Cox regression before and after propensity score matching were used to compare major adverse cardiac and cerebrovascular events (MACCE), including myocardial infarction (MI), stroke, death, and repeat revascularization. During follow-up, MACCE occurred in 20.5% of patients in the PCI group and 8.6% of patients in the CABG group (hazard ratio (HR): 3.263, 95% confidence interval (CI): 1.379 to 7.722, p=0.007). Repeat revascularization occurred more frequently in the PCI group (18.9% vs. 3.7%, respectively, HR: 6.968, 95% CI: 2.036 to 23.842, p=0.002). There were no significant differences in the other endpoints. After propensity score matching, no conclusions were modified. In young patients with LM or three-vessel disease, PCI showed a higher incidence of MACCE, which was mainly driven by repeat revascularization. However, this did not translate into hard endpoint differences. Therefore, PCI is an alternative treatment to CABG in young patients with complex CAD.